COMMUNITY ENGAGEMENT/OUTREACH CORE
D Paul, Vice Chair
Meharry Medical Collegecity: Nashville country: United States (us)
Abstract: Under the leadership of Dr. Paul Juarez, the CEOC will establish the Academic/Community Partnership to Reduce Health Disparities (ACP) to coordinate community involved research, education and outreach activities at MMC, TSU, and Vanderbilt University. Dr. Juarez has considerable experience in conducting community-based participatory research, providing continuing education, training, and outreach to health professionals and community providers, and promoting consumer health education. The ACP will extend efforts begun by the Community Outreach Core (COC) of the Meharry EXPORT Center over the past four years in which the COC established an institutional committee to coordinate community engaged research, education, and outreach across research centers at Meharry (see progress report). It is our hypothesis that engaging community participation in efforts to reduce health disparities in racial/ethnic minority communities will yield new knowledge about the role of individual, community, and macro social level determinants of diseases and public health conditions that will lead to effective interventions and a reduction in health disparities among racial/ethnic minorities. The overall goals of the ACP is to provide a comprehensive, coordinated, and integrated plan of activities organized around defined areas of health disparities to (1) advance community participation in health disparities research arising from diseases and public health conditions that disproportionately affect minority populations; (2) promote the translation and diffusion of evidence-based research findings into clinical and public health practice with our community partners; and (3) coordinate community health education and outreach activities among the three academic partner institutions; and (4) reduce health disparities in Nashville/Davidson County. The ACP will work with existing community engagement cores of other research center at the three institutions communities that promotes a systems framework in addressing health disparities. The ACP initially will invite Core research faculty and members of existing community advisory groups of research, education, and training centers at Meharry, TSU, and Vanderbilt to participate in a coordinated effort to reduce health disparities in Nashville/Davidson County. It also will work to identify and coordinate efforts to address other unmet health needs and areas of health disparities. Community partners will include representatives from public, non-profit and faith based organizations, public schools, and businesses in Nashville/Davidson County. For the purpose of this application, the primary community of interest is underserved minority populations that experience health disparities within Nashville/Davidson County. However, our efforts to engage community in research, education, and outreach will extend beyond the community of interest to include other organizations and individuals that are located in or serve the needs of residents of Nashville/Davidson County. Existing centers at the three institutions with organized community-based research continuing health education, and community education and outreach cores to address areas of health disparities include the U54 Vanderbilt/Meharry Cancer Center (UNPAC), the Urban Partnership Academic Center of Excellence to prevent youth violence (NCCYS), the Title HI Ryan White HIV/AIDS Center for Health Promotion (HTV/AIDS Community Advisory Committee), state model AHEC program (community health professions advisory committee), the Center for Women´s Health, and the .HBCU Wellness Program (obesity/diabetes/chronic diseases), and the Vanderbilt/Meharry CTSA. Dr. Juarez serves on the community engagement and/or advisory groups for each of these major research centers. The ACP also will work with Alignment Nashville, a community-based organization established to increase community participation with the public schools, to coordinate health education and careers in health professions education and training opportunities for middle and high school students in the Nashville Public Schools. 4.4.1. Specific Aims. Specific aims of the Community Engagement and Outreach Core are to 1. Establish the Academic/Community Partnership to Reduce Health Disparities in Nashville/Davidson County. 2. Engage the community to participate in the development, conduct, and evaluation of a health disparities research (e.g. CBPR); 3. Provide continuing education on best practices for addressing health disparities to health care providers who work with medically underserved populations; 4. Recruit disadvantaged and underrepresented minority students at the target middle and high schools in Nashville Public Schools to participate in academic enrichment and support services to better prepare them for careers in health professions;. 5. Host promote community outreach and education to reduce health disparities; and 6. Conduct an annual academic/community health summit on health disparities
Keywords: Acquired Immunodeficiency Syndrome; Address; Advisory Committees; Affect; AIDS/HIV problem; Area; base; Businesses; Cancer Center; career; Centers of Research Excellence; Chronic Disease; Clinical; Communities; community based participatory research; Community Health; Community Health Education; Community Outreach; Community Participation; Continuing Education; County; Development; Diabetes Mellitus; Diffusion; Disadvantaged; Disease; Education and Outreach; effective intervention; Evaluation; evidence base; experience; Faculty; Goals; Health; health disparity; Health education; Health Occupations; Health Personnel; Health Professional; Health Promotion; high school; Historically Black Colleges and Universities; Individual; Institution; interest; Knowledge; Lead; Leadership; medically underserved population; member; middle school; Minority; Minority Groups; Modeling; National Center on Minority Health and Health Disparities; Obesity; outreach; Population; programs; Progress Reports; Provider; public health medicine (field); Public Health Practice; Recruitment Activity; Research; Role; Schools; Services; social; Students; System; Training and Education; Translations; Underrepresented Minority; Universities; Wellness Program; Women`s Health; Work; youth violence prevention
Budget start date: 1-DEC-2010
Budget end date: 30-NOV-2011
5P20MD000516-07_7213 (2011): $186180
Sponsored Links Excellgen http://Excellgen.com
Grants awarded to D Paul
EPIGENETIC CONTROL OF A RAS ACTIVATOR
D Paul
Cornell University Ithacacity: Ithaca country: United States (us)
Grant 5R01CA098597-09 from National Cancer Institute
Abstract: With genome sequence data available in massive abundance, the architecture of the mammalian genome, and the individual variation governing our traits are becoming increasingly understood. The technologies that have produced our sequence data are now being applied to characterize the structure and variation within our epigenome. Epigenetic modifications, which include DNA methylation and a variety of covalent changes to histone tails, are emerging as master regulators that control how sequence information within our genome is utilized. Proper placement and removal of epigenetic marks is essential for normal development, and misplacement of these marks are initiating events for many disease states. A host of enzymes and cofactors that place modifications in the epigenome are known, but almost nothing is known about how they select genomic locations to modify. Understanding how epigenetic states are established and maintained is of fundamental importance to human biology. My lab has identified a naturally occurring DNA sequence that has DNA methylation promoter activity It is both necessary and sufficient to direct the placement of local DNA methylation in mice. No other naturally occurring sequence with this activity has been identified. Just as the dissection of transcriptional promoters facilitated the discovery of transcription mechanisms, the DNA methylation promoter we´ve discovered provides a unique opportunity for elaborating the mechanisms governing placement of this essential epigenetic mark. There are four prominent sequence features within the DNA methylation promoter hypothesized to be important for its function. The four Aims proposed here will reveal their roles in promoting DNA methylation
Keywords: Architecture; Binding (Molecular Function); Coenzymes; Consensus Sequence; Data; Development; Disease; Dissection; DNA Methylation; DNA Sequence; Embryo; Employee Strikes; Epigenetic Process; Event; Excision; Female; G-Quartets; Genetic Transcription; Genome; genome sequencing; Genomics; Germ Lines; Goals; Histones; Human Biology; imprint; Individual; Length; Location; Maintenance; male; mammalian genome; Mammals; Methylation; Modification; Mus; preimplantation; Promotor (Genetics); ras-GRF1; Research; Role; Series; Site; Structure; System; Tail; Technology; tool; trait; Transgenic Mice; Transgenic Organisms; Variant; Work
Project start date: 2003-02-25
Project end date: 2012-07-31
Budget start date: 1-AUG-2011
Budget end date: 31-JUL-2012
PFA/PA: PA-07-070
5R01CA098597-09 (2011): $283822
CX43 PHOSPHORYLATION MODULATES KRAS MEDIATED PANCREAS CANCER PROGRESSION
D Paul, Member
Fred Hutchinson Cancer Research Centercity: Seattle country: United States (us)
Grant 5R21CA149554-02 from National Cancer Institute
Abstract: With an annual incidence and mortality of ~38,000 people, pancreas cancer or pancreatic ductal adenocarcinoma (PDA) is the fourth leading cause of cancer-related death in the United States. The 5-year overall survival for patients with PDA is less than 3%. The gap junction protein connexin43 (Cx43) is a tumor suppressor gene that is expressed in pancreatic ductal cells, the putative site of cancer origin. We have preliminary evidence that Cx43 expression, localization and phosphorylation is dysregulated during pancreas cancer. This proposal focuses on how these events interplay in vivo to affect carcinogenesis of the pancreas. Gap junctions (GJ) are specialized membrane domains that contain channels that allow exchange of small molecules (<1000 Da) including ions, metabolites, and second messengers (e.g., Ca2+ and IP3) between neighboring cells. Connexins, like other junctional proteins, also play critical signaling and growth control roles that are independent of channel function. Very extensive correlative evidence in vivo and cell lines indicates that gap junctional intercellular communication (GJC) and connexin expression regulate proliferation and play key tumor prevention roles. Cx43, by far the most widely expressed connexin (> 34 tissues and 46 cell types), is phosphorylated at multiple serine residues found in the cytoplasmic, C-terminal region. Cx43 phosphorylation can modulate the levels of protein trafficking, stability of the junctional complex, gap junctional communication (GJC) and the interaction with other proteins. We have found that homozygous "knock-in" (KI) of mutant Cx43 bearing an S to A amino acid substitutions at casein kinase 1 (CK1) phosphorylation sites S325, S328 and S330 (termed CK3*) leads to dramatically reduced Cx43-dependent GJC and sustained increased MAPK activity and decreased apoptosis in tissues in response to different acute stimuli. Our collaborator has recently developed the first models of preinvasive and invasive pancreatic ductal adenocarcinoma through the targeted physiologic expression of oncogenic KrasG12D to the pancreas. Resected pancreata demonstrate the full spectrum of preinvasive lesions seen in patients, and the lesions progress histologically over time culminating in fully invasive and metastatic disease. However, like in humans with pancreas cancer, disease progression, symptom presentation and speed of progression varies even in these syngeneic models. We hypothesize that Cx43 phosphorylation is critical for the control of cell growth and is modulated during pancreas carcinogenesis affecting its progression. Consequently, prevention of these regulatory events will result in an altered course of carcinogenesis in pancreas cancer. We hypothesize that Cx43 phosphorylation is critical for the control of cell growth and is modulated during pancreas carcinogenesis affecting its progression. Since drugs that affect Cx43 activity and gap junctional communication are under development, understanding how these factors affect progression could ultimately lead to better treatment of this lethal disease
Keywords: Acute; Affect; Amino Acid Substitution; Animal Model; Animal Models and Related Studies; Animals; Anti-Oncogenes; Antibodies; Antioncogenes; Apoptosis; Apoptosis Pathway; Area; Belief; biological signal transduction; Biology; Body Tissues; c-erbB-1; c-erbB-1 Protein; C-K-RAS; C-terminal; Cancer Biology; Cancer Cause; Cancer Etiology; Cancer Induction; cancer location; cancer progression; cancer site; carcinogenesis; Carcinoma; Casein Kinase 1; casein kinase I; Cell Communication and Signaling; Cell Death, Programmed; cell growth; Cell Line; Cell Lines, Strains; Cell Signaling; Cell to Cell Communication and Signaling; cell type; Cell-Cell Signaling; CellLine; Cells; Cellular Expansion; Cellular Growth; Cessation of life; Characteristics; Communicating Junction; Communication; Complex; Connexin 43; Connexin43; Connexins; cultured cell line; Cx43; Death; Development; Disease; Disease Progression; disease/disorder; Disorder; Disseminated Malignant Neoplasm; DNA Alteration; DNA mutation; Down-Regulation; Down-Regulation (Physiology); Downregulation; drug/agent; Drugs; Ductal Cell; Ductal Epithelial Cell; EGFR; Emerogenes; Epidermal Growth Factor Receptor; Epidermal Growth Factor Receptor Kinase; Epidermal Growth Factor Receptor Protein-Tyrosine Kinase; epithelial carcinoma; Epithelial Neoplasms, Malignant; Epithelial Tumors, Malignant; ERBB Protein; erbB-1; erbB-1 Proto-Oncogene Protein; ERBB1; erbBl; Event; Exhibits; Extracellular Signal-Regulated Kinase Gene; Gap Junction Proteins; Gap Junctions; Gastrointestinal Tract, Pancreas; Gene Alteration; Gene Mutation; gene product; Generalized Growth; Genes, Cancer Suppressor; Genes, Onco-Suppressor; Genetic Alteration; Genetic Change; Genetic defect; Genetic mutation; genome mutation; Growth; heavy metal lead; heavy metal Pb; HER1; Histologic; Histologically; Human; Human, General; in vivo; Incidence; intercellular communication; Intracellular Communication and Signaling; Intracellular Second Messengers; Invasive Lesion; Investigation; Investigators; Ions; K-Ras 2A; K-RAS2A; K-RAS2B; Ki-RAS; Knock-in; Knock-in Mouse; KRAS; KRAS2; KRAS2 gene; L-Serine; Lead; Lesion; Low-resistance Junction; Malignant neoplasm of pancreas; Malignant Pancreatic Neoplasm; Mammals, Mice; Man (Taxonomy); Man, Modern; MAP Kinase Gene; MAPK; Mediating; Medication; Membrane; membrane structure; Metastatic Cancer; Metastatic Malignant Neoplasm; Mice; migration; Mitogen-Activated Protein Kinase Gene; model organism; Modeling; Mortality; Mortality Vital Statistics; mouse model; Murine; Mus; mutant; Mutation; neoplasm progression; neoplastic progression; Nexus; Nexus Junction; Normal Tissue; Normal tissue morphology; Oncogene K-Ras; Oncogene, K-Ras-2; Oncogenes, Recessive; Oncogenes-Tumor Suppressors; Oncogenesis; Oncogenic; oncosuppressor gene; ontogeny; Pancreas; Pancreas Cancer; Pancreas Ductal Adenocarcinoma; Pancreatic; Pancreatic Cancer; Pancreatic Ductal Adenocarcinoma; Pathologist; pathway; Pathway interactions; Patients; Pb element; Pharmaceutic Preparations; Pharmaceutical Preparations; Phosphorylation; Phosphorylation Site; Physiologic; Physiological; Play; Prevention; Protein Kinase CK1; Protein Kinase CKI; Protein Phosphorylation; Protein Trafficking; protein transport; Proteins; proto-oncogene protein c-erbB-1; public health relevance; RASK2; Receptor, EGF; Receptor, TGF-alpha; Receptor, Urogastrone; Receptors, Epidermal Growth Factor-Urogastrone; Research Personnel; Researchers; Resected; Resistance; resistant; response; Role; second messenger; Second Messenger Systems; Second Messengers; Sequence Alteration; Serine; Signal Transduction; Signal Transduction Systems; Signaling; Site; small molecule; social role; Speed; Speed (motion); Stimulus; Survival Rate; Symptoms; Testing; Time; Tissue Growth; Tissues; Traffickings, Protein; Transforming Growth Factor alpha Receptor; tumor; tumor progression; Tumor Suppressing Genes; Tumor Suppressor Genes; tumorigenesis; United States; v-Ki-RAS2 Kirsten Rat Sarcoma 2 Viral Oncogene Homolog
Relevance: We hypothesize that Cx43 phosphorylation is critical for the control of cell growth and is modulated during pancreas carcinogenesis affecting its progression. Since drugs that affect Cx43 activity and gap junctional communication are under development, understanding how these factors affect progression could ultimately lead to better treatment of this lethal disease
Project start date: 2010-02-01
Project end date: 2013-01-31
Budget start date: 1-FEB-2011
Budget end date: 31-JAN-2012
PFA/PA: PA-08-208
5R21CA149554-02 (2011): $3870
COMBINATORIAL CONTRIBUTIONS OF HERITABLE NEUROENDOCRINE VARIATION TO MALEINFERTI
D Paul, Professor Of Biology
College Of William And Marycity: Williamsburg country: United States (us)
Grant 1R15HD068962-01 from Eunice Kennedy Shriver National Institute Of Child Health & Human Development
Abstract: This project will use a unique mammalian study system that was developed specifically to answer questions about heritable neuroendocrine variation that contributes combinatorialy to variation in fertility. Human populations hold large amounts of genetic variation from a great many variable genes that affect complex conditions such as infertility or obesity. Many variable alleles make small or environment-dependent contributions to any given physiological trait, many alleles have cost-benefit trade-offs (and so are negative in some ways, but beneficial in others), some alleles have antagonistic pleiotropic effects (being beneficial for one trait with a cost for another trait), and other alleles have complex epistatic interactions. Recent reviews state that alleles for infertility that have been discovered in men account for a small proportion of cases of infertility. Recent theoretical studies explain why this is expected for such alleles, even in large DNA-sequence studies. Despite these poor results, most studies of heritable causes of infertility continue to be DNA sequence studies. An alternative is to use natural populations to identify and test heritable variation in physiological traits that contribute to fertility and infertility. In my laboratory over the past 15 years, two artificial selection lines from a natural population of white-footed mice (Peromyscus leucopus) have been developed to be either fertile or infertile, respectively, in one environment (short winter-like photoperiod, SD), but fully fertile in another environment (long photoperiod, LD). The selection lines vary heritably and substantially in three neuroendocrine traits that are important regulators of the mammalian reproductive system. The lines also vary heritably in food intake and metabolism, which are known to affect the reproductive system. In this proposal, measurements of values for these traits in a large sample of individuals from an unselected Control line of mice will be used to test for combinatorial contributions of these traits to male fertility and infertility, along with predicted genes-by-environment interactions with photoperiod (SD vs. LD). Additional experiments will test additional, potentially variable neuroendocrine traits to add to an explanatory general linear model for variation in fertility. This proposal is a novel alternative to current research. This study is possible only because of this unique mammalian study system. Success in this project will begin to explain how combinatorial heritable variation from neuroendocrine traits might contribute to infertility in humans. It may be both simpler and more effective to diagnose and treat male infertility using neuroendocrine predictors in mathematical models of combinatorial neuroendocrine variation. This project investigates how heritable variation in multiple traits of the brain, pituitary, and testes combine to cause impaired fertility or infertility in males. A novel experimental system of white-footed mice was developed in order to identify heritable variation in brain, behavior, and hormonal traits related to fertility. This project is intended to help us understand the 30% of cases of male infertility that have no known causes, and to direct future research into infertility
Keywords: Accounting; Affect; Alleles; base; Brain; brain behavior; Clinical Treatment; combinatorial; Complex; cost; Costs and Benefits; Data; Development; Diagnosis; DNA Sequence; Eating; Energy Metabolism; Environment; Fertility; Food Energy; gene environment interaction; Genes; Goals; Gonadal structure; Gonadotropin Hormone Releasing Hormone; Gonadotropins; Hormonal; Hormones; Human; improved; Individual; Infertility; kisspeptin; Laboratories; Linear Models; Luteinizing Hormone; male; Male Infertility; Mammals; mathematical model; Measurement; Measures; men; Metabolism; Modeling; Mus; Nature; Neurons; Neurosecretory Systems; novel; Obesity; Pathway interactions; Peromyscus; Phenotype; Photoperiod; Physiological; Pituitary Gland; Population; Regulation; reproductive; Reproductive system; Research; research study; Rodent; Sampling; Source; Sperm Count Procedure; success; System; Testing; Testis; Theoretical Studies; trait; Variant; Variation (Genetics); White-Footed Mouse
Relevance: This project investigates how heritable variation in multiple traits of the brain, pituitary, and testes combine to cause impaired fertility or infertility in males. A novel experimental system of white-footed mice was developed in order to identify heritable variation in brain, behavior, and hormonal traits related to fertility. This project is intended to help us understand the 30% of cases of male infertility that have no known causes, and to direct future research into infertility
Project start date: 2011-06-15
Project end date: 2014-05-31
Budget start date: 15-JUN-2011
Budget end date: 31-MAY-2014
PFA/PA: PA-10-070
1R15HD068962-01 (2011): $378182
IMPACT OF HIV ON THE T CELL REPERTOIRE
D Paul
University Of California San Franciscocity: San Francisco country: United States (us)
Grant 5K23AI073100-05 from National Institute Of Allergy And Infectious Diseases
Abstract: This K23 proposal is designed to foster my development as a translational researcher in the field of HIV immunology. My proposal is based on a novel assay, called AmpliCot, which I have developed to measure the diversity of the T cell receptor (TCR) repertoire. Although the TCR repertoire is a crucial determinant of the immune response, only now does AmpliCot make it possible to quantitatively measure changes in the repertoire diversity of clinical samples. Under the experienced mentorship of Dr. Joseph M. McCune and Dr. Steven Deeks, and with the guidance of a team of expert consultants in clinical laboratory medicine (Dr. Timothy Hamill), clinical trial design (Dr. Jeffrey Martin), and biostatistics (Dr. Mark Segal), I will obtain the practical knowledge necessary to bring my laboratory investigations to the clinic, launching my career in translational research. HIV infection is known to deplete the body´s number of CD4+ T lymphocytes, but little is known about the effect of infection on the repertoire of TCR specificities. I will use AmpliCot to address the following aims Aim 1 To optimize the AmpliCot assay for use with clinical specimens and to define the normal reference interval for repertoire diversity in healthy adults. Aim 2 To characterize the damage HIV inflicts on TCR repertoire diversity. Aim 3 To assess the degree to which antiretroviral therapy can reconstitute TCR repertoire diversity. The proposed studies are planned in collaboration with two large ongoing cohort studies of HIV-infected individuals at San Francisco General Hospital, thereby facilitating recruitment and retention of subjects, streamlining patient visits, and aiding data analysis. This proposal is a first step towards testing the hypothesis that HIV-infected patients are vulnerable to opportunistic infections because they have lost important TCR specificities and have "holes" in their repertoires. This research could play an important role in helping doctors determine the optimal time to begin treating HIV-infected patients with antiretroviral therapy, and the AmpliCot assay could be used in the future to evaluate the ability of alternative treatments to reconstitute the immune system of HIV-infected patients. The data and skills I will obtain in the K23 award will serve as a foundation for a subsequent prospective study (to be proposed in a future R01 grant application) to investigate whether measurements of lymphocyte receptor diversity can predict risks of opportunistic infections or other clinical events in HIV-infected patients
Keywords: Acquired Immunodeficiency Syndrome; Acute; Address; Adult; Aftercare; Age; alternative treatment; Anti-Retroviral Agents; antiretroviral therapy; Applications Grants; base; Biological Assay; Biometry; Blood; career; CD4 Lymphocyte Count; CD4 Positive T Lymphocytes; Cell Count; Cells; Clinic; Clinical; Clinical Research; Clinical Trials Design; clinically relevant; Cohort Studies; Collaborations; Data; Data Analyses; design; Development; Disease; Ethnicity aspects; Event; experience; falls; Fostering; Foundations; Freezing; Future; Gender; General Hospitals; Genomics; Goals; Highly Active Antiretroviral Therapy; HIV; HIV Infections; human subject; Immune; Immune response; Immune system; Immunocompetence; Immunologic Deficiency Syndromes; Immunology; Individual; Infection; Investigation; Kinetics; Knowledge; Laboratories; Lead; Letters; Link; Lymphocyte; Lymphocyte Count; Measurement; Measures; Mediating; Medicine; Mentored Patient-Oriented Research Career Development Award; Mentorship; Methods; Normal Range; novel; Opportunistic Infections; pathogen; Pathogenesis; Patients; Peripheral; Play; Principal Investigator; Process; programs; Prospective Studies; receptor; reconstitution; Reference Values; Relative (related person); Research; Research Personnel; Risk; RNA; Role; Sample Size; Sampling; San Francisco; skills; Specificity; Specimen; Staging; T-Cell Antigen Receptor Specificity; T-Cell Receptor; T-Cell Receptor Genes; T-Lymphocyte; Techniques; Testing; Time; Translational Research; Visit
Project start date: 2007-05-15
Project end date: 2012-04-30
Budget start date: 1-MAY-2011
Budget end date: 30-APR-2012
PFA/PA: PA-05-143
5K23AI073100-05 (2011): $124200
3K23AI073100-04S1 (2010): $48598
DETERMINANTS OF INFECTIOUS HIV-1 PARTICLE PRODUCTION
D Paul, Professor
Aaron Diamond Aids Research Centercity: New York country: United States (us)
Grant 5R01AI050111-10 from National Institute Of Allergy And Infectious Diseases
Abstract: This is a competing renewal application in which we request support for our studies of retrovirus assembly. In the next grant period, we will address two important mechanistic questions relating to the cell biology of retroviral particle assembly. Specific Aim 1 is to understand how retroviral Gag proteins are targeted to specific sites in various cell types and the purpose of the myristoyl-switch in retarding HIV-1 Gag membrane binding. We will first refine and elaborate our biochemical and real-time live cell imaging assays of retrovirus assembly to determine the route that HIV-1 Gag takes to sites of assembly and egress in various cell types, including primary macrophages. These assays, should enable us to determine how HIV-1 Gag proteins select and move to sites of particle assembly. We will test the hypotheses that active transport pathways, or the intrinsic membrane binding properties of Gag, or both, govern Gag targeting. Additionally, we will determine the consequences for HIV-1 assembly and egress of targeting Gag to distinct cellular membranes in various cell types. Moreover we will test several hypotheses as to why HIV-1 has evolved a mechanism that inhibits membrane binding. Specific Aim 2 is to determine how HIV-1 Vpu affects the targeting of retroviral Gag proteins and promiscuously affects enveloped viral particle release. At present, the mechanism by which Vpu facilitates virus release is almost completely unclear. However our preliminary studies reveal dramatic relocalization of HIV-1 and MLV Gag proteins to the plasma membrane in the presence of Vpu. We will undertake a comprehensive mutagenesis approach to map the determinants of Vpu localization, and to determine the importance of Vpu localization and its effects on Gag localization in the stimulation of virus release virus release. We will also determine the underlying mechanism by which Vpu causes relocalisation of retroviral Gag proteins Finally we will attempt to identify cellular targets and/or cofactors of Vpu´s virus release function using tandem affinity purification. Understanding how HIV-1 and other retroviral Gag proteins select sites with the cell at which to assemble, and how viral and cellular activities influence this process, remains one of the most enigmatic problems in retrovirology. The experiments proposed herein should provide greater clarity as to how Gag proteins behave during assembly in cells
Keywords: Active Biological Transport; Address; Affect; Affinity Chromatography; Binding (Molecular Function); Biochemical; Biological Assay; Cell membrane; cell type; Cells; Cellular biology; cellular imaging; Cellular Membrane; cellular targeting; cofactor; Consequences of HIV; gag Gene Products; Gagging; Grant; HIV-1; Life; macrophage; Maps; Membrane; Mutagenesis; particle; Pathway interactions; Process; Production; Property; research study; Retroviridae; Retrovirology; Route; Site; Testing; Time; Viral; Virus
Project start date: 2001-04-01
Project end date: 2011-09-30
Budget start date: 1-APR-2010
Budget end date: 30-SEP-2011
5R01AI050111-10 (2010): $270106
ENVIRONMENTAL CONTEXT OF HEALTH DISPARITIES
D Paul, Vice Chair
Meharry Medical Collegecity: Nashville country: United States (us)
Grant 3P20MD000516-07S1 from National Institute On Minority Health And Health Disparities
Abstract: This application proposes to further environmental public health research by (1) focusing on health disparities that contribute disproportionately to premature death and morbidity found among poor and racial/ethnic minorities; (2) expanding the definition of environment to include built, social, and policy environments in addition to the natural environment; (3) employing High Throughput Analysis (HTA), spatial analyses, and multi-level analyses to increase our understanding of the relationships between health and environmental factors at the state, county, and sub-county levels, over time; (4) conducting trans-disciplinary training on environmental health disparities; and (5) engaging health disparities communities in the research endeavor through PPGIS and interactive mapping. Three Specific Aims are proposed 1. To establish an environmental health disparities research core that supports analysis of the complex interactions between health disparities and the natural, built, social and policy environments; 2. To promote the use of trans-disciplinary models and analyses to increase knowledge about the complex relationships between health disparities and the physical, built, social and policy environments; 3. To use public participatory geographic information systems (PPGIS) to engage community stakeholders in the use of spatial data and interactive mapping to reduce health disparities. Aims will be carried out by establishing an Environmental Health Disparities Core (EHDC) and integrating EHDC activities into the executive, research, research training, and community engagement cores of the parent p-20 grant. An inter-institutional, trans-disciplinary team of investigators has been recruited to oversee the development of a relational data base and web portal and to analyze relationships between health disparities and environmental factors. Collaborating institutions include the Department of Bioinformatics & Computational Biology, University of Tennessee, Knoxville, School of Public Health and Tropical Medicine, Tulane University, New Orleans, LA, Maryland Institute for Applied Environmental Health, School of Public Health, University of Maryland, and the Universities Space Research Association at Marshall Space Flight Center/National Space Science and Technology Center. Huntsville. AL. Only two of the eleven southeastern ´Stroke Belt´ states currently participate in the CDC´s EPHT initiative. Current EPHT efforts are limited in their approach as they pertain to health disparities. A focus on health disparities will require a more comprehensive, ecological or systems approach that extends the exposure disease paradigm to include the effects of the built, social, and policy environments
Project start date: 2003-09-30
Project end date: 2012-11-30
Budget start date: 21-SEP-2011
Budget end date: 30-NOV-2011
PFA/PA: RFA-MD-11-007
3P20MD000516-07S1 (2011): $279140
MECHANISMS OF ETHANOL-INDUCED NEUROINFLAMMATION AND NEURODEGENERATION
D Paul
University Of Arkansas Med Scis Ltl Rockcity: Little Rock country: United States (us)
Grant 5RC1AA019108-02 from Office Of The Director, National Institutes Of Health
Abstract: There are currently more than 300,000 patients receiving chronic dialysis therapy in the United States, and it is estimated that this population will rapidly rise to over 600,000 patients by 2010 and over 2,000,000 patients by 2030. Life expectancies for dialysis patients are only 1/3 to 1/6 of those of the general population with cardiovascular disease accounting for approximately 50% of mortality. At present, there are no therapies proved to lower the risk for cardiovascular morbidity and mortality in the dialysis population. Conventional cardiovascular risk factors exemplified in the Framingham Study have limited prognostic value in dialysis patients, suggesting that "non-traditional" risk factors influenced by the uremic metabolic milieu may be particularly important. Increased oxidative stress contributes to the pathogenesis of atherosclerosis and uremia has recently been identified as an increased oxidative stress state. Coenzyme Q10, a readily available dietary supplement frequently used as an alternative and complementary therapy is a potent lipophilic antioxidant that couples electron transport to oxidative phosphorylation in the mitochondria. Coenzyme Q10 administration has been associated with significant clinical benefits in the treatment of congestive heart failure and improves endothelial function in patients with type II diabetes mellitus and preserved kidney function. Plasma concentrations of coenzyme Q10 appear to reflect net overall metabolic demand, and our preliminary data demonstrates that plasma coenzyme Q10 levels are depressed in hemodialysis patients, suggesting that coenzyme Q10 may be an ideal antioxidant for these patients. The central hypothesis of this application is that administration of coenzyme Q10 as a targeted antioxidant therapy will be safe and well tolerated, and will ameliorate the excessive oxidative stress burden in hemodialysis patients. This in turn will lead to improvements in surrogate markers for cardiovascular risk. We propose to test this hypothesis through the following Specific Aims A.1. To determine the safety and tolerability of different doses of dietary supplement coenzyme Q10 in hemodialysis patients. A.2. To test the effect of the dietary supplement coenzyme Q10 on biomarkers of oxidative stress, systemic inflammation and endothelial function in hemodialysis patients. Patients receiving chronic dialysis therapy have a high death rate due to cardiovascular disease. This project seeks to test whether administration of coenzyme Q10, a readily available dietary supplement, could result in clinical benefits for dialysis patients. This study is also designed to test whether administration of coenzyme Q10 to dialysis is safe and well-tolerated
Keywords: Accounting; Address; Aldehydes; Anabolism; antioxidant therapy; Antioxidants; Atherosclerosis; atorvastatin; base; Biochemistry; Biological Assay; Biology; biomarker; Cardiac Death; Cardiovascular Diseases; cardiovascular pharmacology; cardiovascular risk factor; Cardiovascular system; Chronic; Chronic Kidney Failure; Clinical; Clinical Trials; Coenzyme Q10; Complementary therapies; Congestive Heart Failure; Couples; Data; Death Rate; depressed; design; diabetic; Dialysis patients; Dialysis procedure; Dietary Supplements; Dose; Electron Transport; Future; General Population; Hemodialysis; Hydroxymethylglutaryl-CoA reductase; Hydroxymethylglutaryl-CoA Reductase Inhibitors; improved; Inflammation; Isoprene; Kidney; Laboratories; LDL Cholesterol Lipoproteins; Lead; Life Expectancy; Lipid Peroxidation; Medical; Metabolic; Metabolism; Mitochondria; Modeling; Morbidity - disease rate; Mortality Vital Statistics; multidisciplinary; Myocardial Infarction; Non-Insulin-Dependent Diabetes Mellitus; novel strategies; nutrition; Oxidation-Reduction; Oxidative Phosphorylation; Oxidative Stress; Pathogenesis; Patients; Plasma; Population; Population Study; prognostic; public health relevance; Renal function; Risk Factors; Risk Reduction; Safety; Secondary to; sound; stroke; Sulfhydryl Compounds; Surrogate Markers; Testing; Therapeutic; United States; Uremia
Relevance: Patients receiving chronic dialysis therapy have a high death rate due to cardiovascular disease. This project seeks to test whether administration of coenzyme Q10, a readily available dietary supplement, could result in clinical benefits for dialysis patients. This study is also designed to test whether administration of coenzyme Q10 to dialysis is safe and well-tolerated
Project start date: 2009-09-25
Project end date: 2011-08-31
Budget start date: 1-SEP-2010
Budget end date: 31-AUG-2011
PFA/PA: RFA-OD-09-003
5RC1AA019108-02 (2010): $499999
NEUROSCIENCE RESEARCH CENTER CORE FACILITY AT UAMS
D Paul
University Of Arkansas Med Scis Ltl Rockcity: Little Rock country: United States (us)
Grant 5P30NS047546-05 from National Institute Of Neurological Disorders And Stroke
Abstract: The University of Arkansas for Medical Sciences (UAMS) seeks an NINDS Institutional Center Core Grant to Support Neuroscience Research. Neuroscience research is a major contributor to the recent growth of extramurally supported funding at UAMS, with a significant number of principal investigators (PIs) funded by the NINDS. The leadership of UAMS recognizes that having a greater number of NINDS-funded investigators will benefit the entire campus. One mechanism to achieve this increase is to establish a centrally located resource for all neuroscientists at UAMS, providing access to sophisticated equipment, trained technical staff, and the research expertise of established NINDS funded investigators. The long-term goal of this Neuroscience Center is to establish a facility that will foster interactions between members of the UAMS neuroscience community, leading to an increase in competitive research grants and the development of thematic program research. It is expected that creation of this center will serve as a recruiting tool to attract new faculty members with research interests in neuroscience, further expanding neuroscience research at UAMS. To achieve these goals the present proposal focuses on the development of three Cores 1) Animal Surgery and Animal Model Development, 2) Histology and Image Analysis, and 3) Biochemistry, Cell and Molecular Biology. Four Specific Aims address the manner in which the proposed Core facilities will meet the present and future demands of NINDS-funded investigators, thereby ensuring continued productivity and expansion of research directions. Aim 1 is to provide a centralized facility for the three proposed Cores to enhance the research productivity of NINDS-funded investigators so they remain competitive in their research. Aim 2 is to encourage new interactions between NINDS-funded investigators by creating an environment conducive to conducting interdisciplinary neuroscience research. PIs with qualifying projects will have regular opportunities to share ideas and build collaborative relationships when a common Core facility is available. Aim 3 is to enhance the research expertise of neuroscientists at UAMS by providing specialized research resources currently unavailable to most investigators on campus. The application of new research techniques will enhance the present and future research objectives of funded researchers. Aim 4 is to increase the number of NINDS-funded investigators at UAMS by encouraging the interaction of neuroscientists currently without NINDS-funding with PIs conducting qualifying research. As neuroscience researchers continue working together to build strength in critical areas, the NINDS will benefit by working with institutions such as UAMS to develop state of the art, contiguous Core facilities that will support growth and integration of the campus-wide neuroscience community
Project start date: 2004-08-15
Project end date: 2011-07-31
Budget start date: 1-AUG-2008
Budget end date: 31-JUL-2011
PFA/PA: PAR-02-059
5P30NS047546-05 (2008): $600294
DNA METHYLATION IN ALZHEIMER?S DISEASE AND NORMALLY AGED BRAIN
D Paul
Sun Health Research Institutecity: Sun City country: United States (us)
Grant 5R01AG036400-03 from National Institute On Aging
Abstract: The proposed research represents the coordination of a number of existing facilities and resources aimed at defining the role(s) of DNA methylation in the molecular pathology of Alzheimer´s disease. We will integrate the expertise of the Coleman/Rogers lab in Alzheimer´s disease with the expertise of the Laird lab in DNA methylation to identify DNA methylation sites in a genome wide study of an affected (temporal neocortex) and a minimally affected (cerebellum) brain region of Alzheimer´s disease (AD) and non demented control cases. The 550 brains to be utilized in this proposed study will come from the comprehensive and outstanding Brain Bank at Sun Health Research Institute. Since promoter DNA methylation only relates to the potential for gene expression, it may not be a very good predictor of gene expression differences. Consequently, we will determine the relationship between specific sites of DNA methylation and altered expression of specific genes in AD by correlating our DNA methylation data with an existing set of Affymetrix data on gene expression in ~100 normal and AD brains. Finally, to specify cell classes affected we will conduct combined immunohistochemistry and in situ htbridization studies to define cell classes affected by specific DNA methylation/expression changes. A successful attack on Alzheimer´s disease requires two components 1) early diagnosis and 2) effective treatment that will significantly delay or halt disease progression. The genome wide study of DNA methylation in Alzheimer´s disease proposed here will elucidate and extend the previously under appreciated role of a general molecular mechanism in the basic biology of AD and will lead to earlier diagnosis and new approaches to treatment of Alzheimer´s disease
Keywords: Affect; aging brain; Alzheimer`s Disease; Biology; Brain; Brain region; case control; Cells; Cerebellum; Data; Disease; Disease Progression; DNA Methylation; Early Diagnosis; effective therapy; Gene Expression; Genes; genome wide association study; Health; Immunohistochemistry; In Situ; Lead; Molecular; molecular pathology; Neocortex; novel strategies; Promotor (Genetics); Research; Research Institute; Resources; Role; Site; The Sun; To specify
Relevance: A successful attack on Alzheimer´s disease requires two components: 1) early diagnosis and 2) effective treatment that will significantly delay or halt disease progression. The genome wide study of DNA methylation in Alzheimer´s disease proposed here will elucidate and extend the previously under appreciated role of a general molecular mechanism in the basic biology of AD and will lead to earlier diagnosis and new approaches to treatment of Alzheimer´s disease
Project start date: 2009-09-15
Project end date: 2014-08-31
Budget start date: 1-SEP-2011
Budget end date: 31-AUG-2012
PFA/PA: RFA-RM-08-017
5R01AG036400-03 (2011): $388536
Sponsored Links Excellgen http://Excellgen.com
PHYSICAL AND SOCIAL ENVIRONMENTAL FACTORS IN ADULT ASTHMA OUTCOMES
D Paul
University Of California San Franciscocity: San Francisco country: United States (us)
Grant 5R01ES010906-10 from National Institute Of Environmental Health Sciences
Abstract: The proposed project will prospectively investigate physical and social environmental exposures as determinants of Health-Related Quality of Life (HRQOL) in adult asthma and rhinitis, focusing on the role of severity of disease as a principal mediator between the physical and social environment and HRQOL. The proposed study, a competitive renewal of an R01 near completion, builds upon our current research elucidating multifactorial models of physical and social environmental risks. The specific study aims are 1.] Delineate the specific pathways linking physical and social environmental exposures to disease-specific and general HRQOL in adult asthma and rhinitis, including mediation by changing disease status overtime; 2.] Identify specific risk factors, such as personal socioeconomic status, that modify the impact of these environmental factors on disease severity or the effect of severity on HRQOL in adult asthma and rhinitis. To accomplish these goals, we will assemble a prospective cohort of 636 adults, merging our ongoing asthma-rhinitis cohort (the basis of the current R01) with a second cohort of adults with severe asthma established as part of a methodologically similar study. Subjects will undergo 3 waves of structured telephone interviews over the study period to ascertain environmental exposures, disease severity, and HRQOL. Two home visits will be conducted in a subset of subjects (n=380) to assess the home environment (including measurements of dust antigen, airborne particulate, and dampness), measure pulmonary function and exhaled fraction of nitric oxide, and collect biological samples to measure secondhand smoke exposure. We will also carry out geocoding for linkage to external data sets for U.S. Census information, traffic density, and levels of ambient air pollutants. We will test predictive models of physical and social environmental factors, including measures of indoor air quality, ambient pollution, and neighborhood and community status, estimating the longitudinal effect of these factors on HRQOL. We will assess the role of disease severity as a mediating factor in the causal pathway leading from environmental exposures to changes in HRQOL; we will also assess selected individual factors that may be effect modifiers of this relationship, especially personal socioeconomic status (SES). Public Health Importance In order to understand the complex web of factors influencing airway disease outcomes from the perspective of persons with disease, there is a critical need for an analytic approach that can test the effect of multiple combined environmental risk factors on HRQOL, assess the mediating role of disease severity, and take into account individual effect modifiers. The long- term goal of our study is to identify modifiable factors and subgroups at high risk in order to improve HRQOL among adults with asthma. The proposed project builds upon our success in the current R01 by testing more specific pathways in the context of our accumulated experience to date
Keywords: Accounting; Adult; Adult asthma; Affect; Air Pollutants; airway inflammation; Antigens; Asthma; base; Biological; Censuses; cohort; Communities; Complex; Data Set; density; Diagnosis; Disease; Disease Outcome; Dust; Economics; Environment; Environmental Exposure; Environmental Risk Factor; Event; Exhalation; experience; Exposure to; Gender; Generic Drugs; Genetic; Goals; health related quality of life; Health Services Accessibility; Health Status; high risk; Home environment; Home visitation; Hospitalization; House Call; improved; Individual; Indoor Air Quality; Intensive Care; Internet; Life; Link; Measurement; Measures; Mediating; Mediation; Mediator of activation protein; Modeling; Neighborhoods; Nitric Oxide; Outcome; Outcome Study; Particulate; Pathway interactions; Persons; Physical environment; physical model; Physicians; Pollution; predictive modeling; programs; prospective; public health medicine (field); pulmonary function; Quality of life; Research; Research Personnel; Respiratory physiology; Rhinitis; Risk; Risk Factors; Role; Sampling; Severities; Severity of illness; Smoke; social; Social Environment; Socioeconomic Status; Specificity; Structure; Subgroup; success; Symptoms; Telephone Interviews; Testing; Theoretical model; trafficking; Variant; Work
Project start date: 2000-10-01
Project end date: 2012-06-30
Budget start date: 1-JUL-2011
Budget end date: 30-JUN-2012
5R01ES010906-10 (2011): $464916
NCMHD HEALTH DISPARITIES RESEARCH CENTER OF EXCELLENCE AT MEHARRY
D Paul, Vice Chair
Meharry Medical Collegecity: Nashville country: United States (us)
Grant 3P20MD000516-07S2 from National Institute On Minority Health And Health Disparities
Abstract: The overarching goals of the Center are to (1) demonstrate the efficacy of a systems framework for conducting health services research that address multi-level determinants of health disparities among racial/ethnic minorities; (2) provide inter-disciplinary mentoring, support, and training of research scientists in conducting health disparities research designed to prevent or delay the onset of diseases and conditions that disproportionately affect minority health; (3) establish academic and community partnerships to develop, implement, translate, and disseminate discoveries from health disparities research into real world, clinical, public health, and public policy practice; and (4) reduce health disparities among racial/ethnic minorities in Nashville/Davidson County. This application builds upon the unique institutional mission of Meharry, existing collaborative relationships with investigators at Tennessee State University and Vanderbilt University, our commitment to community engagement in research, accomplishments of our past EXPORT Center grant, and the new Meharry institutional research plan. For this application, the name of the Meharry EXPORT Center will be changed to the NCMHD Health Disparities Research Center of Excellence at Meharry (HDRCOE or the "Center") to reflect changes in the current RFA and refinement of our mission. The Center will have four cores Administration, Research, Research Training and Community Engagement. Five specific aims have been identified to guide efforts of the NCMHD Health Disparities Research Center of Excellence (HDRCOE or the "Center") in accomplishing its overarching goals 1 Establish an inter-institutional, inter-disciplinary, NCMHD health disparities research center of excellence at Meharry Medical College (MMC); 2 To conduct inter-disciplinary, health disparities research using a systems framework that will advance our understanding of the interrelationships between human factors, community context, and macro social forces as determinants in the development, progression, prevention, and control of disease and public health conditions that lead to health disparities across the lifespan. 3 To train students, residents, and scholarly productivity on health disparities. 4 To increase exchanges of knowledge about health disparities among faculty, students, and community participants about culturally sensitive diagnostic, prevention, treatment, and policy interventions for preventing, delaying, and managing the onset and progression of conditions and diseases that disproportionately affect health outcomes of racial/ethnic minorities and other medically underserved and at-risk populations. 5 To develop an administrative infrastructure that supports inter-institutional community participation in health disparities research across the three institutions
Keywords: Abnormal coordination; Achievement; Address; Adult; Affect; Appointment; Area; Behavioral; behavioral health; Cancer Center; career; Centers of Research Excellence; Charge; Clinical; clinical care; Clinical Research; Clinical Sciences; Clinical Trials; Cohort Studies; Collaborations; college; Commit; Communities; Community Health; Community Health Education; community organizations; Community Outreach; Community Participation; Community Practice; Consensus; Country; County; Data; Data Analyses; data management; Data Set; Dental Students; design; Development; Diagnostic; Discipline; Disease; disorder control; disorder prevention; Educational aspects; Educational Curriculum; Educational process of instructing; Educational workshop; Effectiveness; empowerment; Enrollment; Ensure; Evaluation; experience; Faculty; falls; Farming environment; Fostering; Frustration; Goals; graduate medical education; graduate student; Grant; Health; health disparity; Health Personnel; Health Professional; Health Promotion; Health Sciences; Health Services; Health Services Research; Health Status; Healthy People 2010; HIV; Hour; Human; Human Development; Human Resources; improved; Individual; Institution; interdisciplinary collaboration; Interdisciplinary Study; interest; Intervention; Investigation; Investments; Journals; Knowledge; Lead; Leadership; Learning; Link; Longevity; Master of Science; Measures; Medical; Medical Education; medical schools; medical specialties; Medical Students; medically underserved; member; Mentors; Methods; Minority; Minority Groups; minority health; Mission; Modeling; Names; National Center on Minority Health and Health Disparities; Nursing Faculty; Onset of illness; Outcome; Outcomes Research; outreach; Participant; Pathway interactions; Persons; Policies; Population; population health; Populations at Risk; portability; Postdoctoral Fellow; prevent; Prevention; Prevention education; Preventive Medicine; Primary Health Care; Process; Productivity; Professional Education; programs; public health medicine (field); Public Health Practice; Public Policy; racial and ethnic; Readiness; Recording of previous events; Relative (related person); Research; Research Activity; research and development; Research Design; Research Infrastructure; Research Personnel; Research Project Grants; Research Training; Residencies; Resources; Role; Rotation; Scientist; Series; skills; social; social health determinants; Structure; Students; success; System; Tennessee; tool; Training; Training Activity; Training and Infrastructure; Training Programs; Training Support; Translating; Translational Research; trend; Trust; Underrepresented Minority; Universities; Vision; Women`s Health; Work; Writing
Project start date: 2003-09-30
Project end date: 2013-11-30
Budget start date: 23-SEP-2011
Budget end date: 30-NOV-2011
PFA/PA: RFA-MD-08-004
3P20MD000516-07S2 (2011): $135609
3P20MD000516-07S3 (2011): $29195
STRUCTURE FUNCTION STUDIES OF RYR1 IN A MYOGENIC KNOCKOUT
D Paul, Professor
Brigham And Women´s Hospitalcity: Boston country: United States (us)
Grant 5R01AR043140-14 from National Institute Of Arthritis And Musculoskeletal And Skin Diseases
Abstract: The long-term goal of the proposed research is to identify the key regions within the primary sequence of theCa2+ release channel (ryanodine receptor; RyR1) of mammalian skeletal muscle sarcoplasmic reticulum (SR) which determine 1. Sensitivity to physiologic activating and inhibiting cations, and 2. The ability of the channel to sense physiologic changes in local redox potential. Hypothesis I Structural determinants of the cation activator and inhibitor sites are coordinated by spatially separated amino acids within the RyR1 linear sequence. Conserved negative charges within one or more of five cytoplasmic domains in the C-terminal -1000 amino acids of RyR1 coordinate cation binding and activation, whereas it appears that interactions between the C-terminal and central domains of RyR1, especially sequences encompassing difference regions D1 & D3, are needed to engage proper cation inhibition of RyRI. Specific Aim 1. Define the structural determinants in RyR1 that influence its binding and sensitivity to Ca2+ and Mg2+ using site-specific mutations of C-terminal truncated channels and full length RyR1 along with RyR1/RyR3 chimera. Specific Aim 2. Define the changes in cation binding constants and the activation energy (Ea) associated with channel activation for RyR1s with mutations within their cation binding motifs. Specific Aim 3. To define how altered cation regulation of RyR1 impacts excitation-contraction coupling and excitation-coupled Ca2+ entry. Hypothesis II Hyper-reactive sulfhydryls in the primary structure of RyR1 constitute a trans- SR redox gradient sensor. Specific Aim 1. To mutate the six hyper-reactive thiols that we have discovered in RyR1 and define their contribution to trans SR redox sensing behavior. Specific Aim 2. To establish mechanistic links between redox state and the ability of cations to regulate RyR1 activity. The proposed studies will provide new information concerning the molecular mechanisms by which microsomal calcium channels residing within the SR/ER membrane are regulated by two important physiological parameters, cations (Ca2+and Mg2+) and cellular redox state. It is likely that these mechanisms are conserved among the multi-member family of Ca2+ release channels and may be significant in identifying new clinical targets for the prevention and treatment of injury resulting from oxidative stress
Keywords: Address; Amino Acids; Behavior; Binding (Molecular Function); Binding Sites; Biochemical; Biological; Budgets; C-terminal; cadmium ion; Calcium Channel; Cations; Cells; Charge; Chimera organism; Clinical; Coupling; Cysteine; Cytoplasmic Tail; EF Hand Motifs; Family member; Fluorescence; Funding; Goals; inhibitor/antagonist; Injury; Knock-out; Length; Link; Measures; Membrane; Modeling; Molecular; Muscle Cells; Muscle Fibers; Mutate; Mutation; Oxidation-Reduction; Oxidative Stress; Physiological; Prevention; Property; protein protein interaction; Regulation; Research; Ryanodine; Ryanodine Receptor Calcium Release Channel; Ryanodine Receptors; RyR1; RyR3; Sarcoplasmic Reticulum; sensor; Site; skeletal; Skeletal muscle structure; Structure; Sulfhydryl Compounds; Techniques
Project start date: 1996-04-14
Project end date: 2012-03-31
Budget start date: 1-APR-2010
Budget end date: 31-MAR-2012
5R01AR043140-14 (2010): $469416
MAGNESIUM NMR OF DNA REPAIR PROTEINS
D Paul, Senior Staff Scientist
Battelle Pacific Northwest Laboratoriescity: Richland country: United States (us)
Grant 5R01EB003893-04 from National Institute Of Biomedical Imaging And Bioengineering
Abstract: The chemistry central to the function of the DNA repair proteins apurinic/apyrimidic endonuclease 1 (Ape1) and polymerase beta (Pol beta) is the chemistry of water activated by a magnesium ion. These proteins are key constituents of the base excision repair (BER) pathway, a process that plays a critical role in preventing the cytotoxic and mutagenic effects of most spontaneous, alkylation and oxidative DNA damage. The proposed research represents a novel application of low temperature (10K) solid-state 25Mg NMR applied to DNA repair proteins and their model systems. The principal aim for the proposed research is to establish a relationship between 25Mg magnetic resonance parameters (quadrupole coupling constants, shielding tensors and their relative orientations) and the structure/function relationships for known Mg-DNA repair proteins and extrapolating those relationships to other Mg-dependent DNA repair proteins where the role of the metal is less defined. These experiments are performed in concert with ab initio electronic structure calculations. The combination of these two methods provides a firm basis for the "chemical" understanding of the role these metals play in their respective proteins. Additionally, triple resonance experiments (1H, [15N or 31P], 25Mg) are proposed to facilitate the identity of neighboring ligands and the determination of selective distances between the magnesium and the substrates. The results of the entirety of these experiments are critical for the mechanistic understanding of the role the metal plays in these critical DNA repair proteins
Keywords: 2-Phenyl-2-Ene-Benzopyran-4-One Compounds; Active Sites; Address; Agreement; Alkaline Phosphatase; alkaline phosphomonoesterase; Alkylation; base; Base Excision Repairs; Binding; Binding (Molecular Function); Biological Models; Chemical Models; chemical structure function; Chemicals; Chemistry; clinical data repository; clinical data warehouse; cold temperature; Complex; computational modeling; computational models; computational simulation; computer based models; Computer Simulation; computerized modeling; Computerized Models; computerized simulation; Coupling; cytotoxic; Data Banks; Data Bases; data repository; Databank, Electronic; Databanks; Database, Electronic; Databases; Deoxyribonucleic Acid; Dependence; DNA; DNA Base Excision Repair; DNA Binding; DNA Binding Interaction; DNA repair protein; electronic structure; endonuclease; Environment; experiment; experimental research; experimental study; flavone; Flavones; Frequencies (time pattern); Frequency; gene product; glycerophosphatase; Goals; H+ element; Hydrogen Ions; Hydrogen Oxide; hydroxide; Hydroxide Ion; Hydroxides; hydroxyl ion; improved; in silico; interest; Investigators; Ions; Ligand Binding; Ligands; low temperature; Magnesium; magnesium ion; Magnetic Resonance; Maps; Mathematical Model Simulation; Mathematical Models and Simulations; Measures; Metal Binding Site; Metals; method development; Methods; Metric; Mg element; Mg++ element; Model System; Modeling; Models, Biologic; Models, Computer; Molecular; Molecular Interaction; mutant; N element; N2 element; Nitrogen; NMR Spectroscopy; Nomenclature; novel; nuclear magnetic resonance spectroscopy; Orthophosphoric-monoester phosphohydrolase (alkaline optimum); oxidative DNA damage; P element; pathway; Pathway interactions; Phosphorus; Play; Polymerase; Preparation; prevent; preventing; Process; programs; Programs (PT); Programs [Publication Type]; protein complex; Proteins; Protons; relational database; Relative; Relative (related person); Repairs, Base Excision; Research; Research Personnel; research study; Researchers; Role; salt; Sampling; Science of Chemistry; Simulation, Computer based; Site; social role; Sodium Chloride; Sodium chloride (NaCl); solid state; solid state nuclear magnetic resonance; Spectroscopy; Spectroscopy, NMR; Spectrum Analyses; Spectrum Analysis; stoichiometry; structure function relationship; Structure-Activity Relationship; System; System, LOINC Axis 4; Temperature; Testing; theories; triphosphate; tripolyphosphate; Variant; Variation; virtual simulation; Water
Project start date: 2006-08-17
Project end date: 2011-06-30
Budget start date: 1-JUL-2009
Budget end date: 30-JUN-2011
5R01EB003893-04 (2009): $585810
A CHARACTERIZATION OF BACULOVIRUS EARLY GENE EXPRESSION
D Paul
University Of Wisconsin Madisoncity: Madison country: United States (us)
Grant 5R01AI025557-20 from National Institute Of Allergy And Infectious Diseases
Abstract: The baculoviruses are large DNA viruses distinguished by their prolific multiplication in host insects. Due to their unique properties, these pathogens are used as vectors for foreign gene expression, potent biological insecticides, and gene transfer vehicles. To ensure their replicative success, baculoviruses use novel mechanisms that expedite viral gene expression and suppress host defenses, including cell death by apoptosis. It is the long term goal of this project to investigate the molecular interactions between baculoviruses and their host cell by defining the mechanisms by which Autographa californica nucleopolyhedrovirus (AcMNPV) regulates gene expression and modulates the host apoptotic response. In an integrated series of experiments that use biochemical, genetic, and cell biology approaches, the molecular mechanisms by which early AcMNPV transcriptional regulators accelerate replication will be investigated. The mechanism of enhancer-mediated transcriptional activation by the immediate early transregulator IE1 will be determined by using new loss-of-function mutations and dominant inhibitors. These dominant inhibitors will be used in combination with interfering RNA (RNAi) to define the roles of IE1 during infection, including the induction of apoptosis. AcMNPV-infected insect cells will be used as a powerful yet convenient system to define the molecular signals by which DNA viruses trigger apoptosis. Capitalizing on the finding that AcMNPV induces widespread apoptosis in cultured Drosophila melanogaster cells, we use dominant inhibitors, RNAi, and the baculovirus apoptotic suppressors (p35, p49, and iap) to investigate the viral and cellular apoptotic factors in this model organism for which many of the cell death components are known. These studies will be complemented by a characterization of the molecular signals (viral and host) responsible for nodavirus-induced apoptosis in Drosophila. Collectively, these studies are expected to provide important insight into the highly conserved pathways of apoptosis, which contribute significantly to the pathogenicity of human viruses. In addition, the molecular mechanisms by which baculoviruses regulate early gene transcription are directly relevant to insect vectors of human disease
Keywords: Animal Model; Apoptosis; Apoptotic; Baculoviruses; Binding (Molecular Function); Biochemical Genetics; Biological; Cell Death; Cell Line; Cells; Cellular biology; Complement; DNA Viruses; Dominant-Negative Mutation; Drosophila genus; Drosophila melanogaster; Early Gene Transcriptions; Enhancers; Ensure; Event; Gene Expression; Gene Transfer; Genes; Goals; Host Defense; human disease; Human Virus; in vivo; Induction of Apoptosis; Infection; inhibitor/antagonist; Insect Vectors; Insecta; Insecticides; insight; loss of function mutation; Mediating; Molecular; novel; Nucleopolyhedrovirus; pathogen; Pathogenicity; Process; Property; Range; Regulation; research study; response; RNA; RNA Interference; Role; Series; Signal Transduction; success; System; Trans-Activation (Genetics); Transcriptional Activation; Transcriptional Regulation; vector; Viral; Virus
Project start date: 1988-02-01
Project end date: 2010-02-28
Budget start date: 1-MAR-2008
Budget end date: 28-FEB-2010
5R01AI025557-20 (2008): $299435
5R01AI025557-19 (2007): $305342
5R01AI025557-18 (2006): $314573
DISCOVERY AND MECHANISM OF ANTIRETROVIRAL FACTORS
D Paul, Professor
Aaron Diamond Aids Research Centercity: New York country: United States (us)
Grant 2R01AI064003-06 from National Institute Of Allergy And Infectious Diseases
Abstract: The ancestors of modern organisms were colonized by viruses that were very rarely beneficial, and often deleterious to the host. Inevitably, hosts adapted to limit viral replication, and in so doing evolved various autonomous antiviral defense mechanisms. One of these is TRIM5, which is multimeric protein that recognizes incoming retroviral capsids, but its precise mechanisms of action are unclear. In specific aim 1 we will develop new techniques to understand what happens to various incoming components of retroviral particles as they complete the early steps of their life cycle. These assays will very likely illuminate the mechanisms by which TRIM5 proteins and other antiretroviral factors exert their inhibitory effects. Moreover, these techniques are likely to have a wide range of applications in the study of these steps of the retroviral life cycle. In specific aim 2 we will derive TRIM5-resistant HIV-1 capsids that will identify residues on the surface of the HIV-1 capsid that are targeted by TRIM5 and will serve a useful tools to probe the mechanism of action of TRIM5. Additionally these capsids will be used for generating simian-tropic HIV- strains that can replicate in rhesus macaques. In addition to known antiretroviral proteins like TRIM5, there are several reasons to think that there are more, perhaps many more, antiviral host defense genes in to be discovered. In specific aim 3, we will attempt to identify such genes by constructing focused, arrayed cDNA libraries, each consisting of tens to hundreds of candidate genes, that either (i) have key properties exhibited by known antiretroviral genes or (ii) are selectively expressed in cells that exhibit constitutive or induced resistance to HIV-1 or SIV infection. These genes libraries will be arrayed each gene will be individually tested for its ability to inhibit retrovirus replication. Humans and laboratory animals have an array of antiviral defense mechanisms, of which we have only a rudimentary understanding. Identifying and understanding the mechanism of action of antiretroviral gene products could lead to completely new chemotherapeutic strategies for tackling infectious diseases, including AIDS. In addition, understanding species-specific variation and engineering resistance to antiretroviral genes could facilitate the development of animal models of human retroviral infection
Keywords: Acquired Immune Deficiency; Acquired Immune Deficiency Syndrome; Acquired Immune Deficiency Syndrome Virus; Acquired Immuno-Deficiency Syndrome; Acquired Immunodeficiency Syndrome; Acquired Immunodeficiency Syndrome Virus; AIDS; AIDS Virus; Animal Model; animal model development; Animal Models and Related Studies; Animals; anti-retroviral; Anti-Retroviral Agents; anti-retroviral resistance; anti-retroviral resistant; antiretroviral; Antiretroviral Agents; Antiretroviral resistance; Antiretroviral resistant; Antiviral Agents; Antiviral Drugs; Antivirals; ARV resistance; ARV resistant; Assay; base; Base Sequence; Bioassay; Biologic Assays; Biological Assay; Candidate Disease Gene; Candidate Gene; Capsid; cDNA Library; cell type; Cells; coat (nveloped virus); Communicable Diseases; Data; Defense Mechanisms; Engineering; Engineerings; Event; Exhibits; Friend virus susceptibility 1; Funding; Fv-1 protein; Fv1; Fv1 protein; gene discovery; Gene Library; gene product; Genes; genetic library; Goals; Grant; heavy metal lead; heavy metal Pb; high reward; high risk; HIV; HIV-1; HIV-I; HIV1; Host Defense; HTLV-III; Human; Human immunodeficiency virus 1; Human Immunodeficiency Viruses; human T cell leukemia virus III; human T lymphotropic virus III; Human T-Cell Leukemia Virus Type III; Human T-Cell Lymphotropic Virus Type III; Human T-Lymphotropic Virus Type III; Human, General; Immunodeficiency Virus Type 1, Human; Immunologic Deficiency Syndrome, Acquired; Infection; Infectious Disease Pathway; Infectious Diseases; Infectious Diseases and Manifestations; Infectious Disorder; inhibitor; inhibitor/antagonist; Interferon Type I; Intervention; Intervention Strategies; interventional strategy; Laboratory Animals; LAV-HTLV-III; Lead; life course; Life Cycle; Life Cycle Stages; living system; Lymphadenopathy-Associated Virus; Macaca; Macaca mulatta; Macaque; Man (Taxonomy); Man, Modern; Methods and Techniques; Methods, Other; model organism; Modeling; Molecular Biology, Nucleic Acid Sequencing; mutant; nucleic acid sequence; Nucleic acid sequencing; Nucleotide Sequence; Organism; particle; Pb element; Pharmaceutical Agent; Pharmaceuticals; Pharmacologic Substance; Pharmacological Substance; Physiologic; Physiological; prevent; preventing; Property; Property, LOINC Axis 2; Proteins; psychological defense mechanism; public health relevance; Relative; Relative (related person); Resistance; resistance to anti-retroviral; resistance to antiretroviral; resistance to ARV; resistant; resistant to anti-retroviral; resistant to antiretroviral; resistant to ARV; Retroviridae; Retroviruses; Rhesus; Rhesus Macaque; Rhesus Monkey; selective expression; selectively expressed; SEQ-AN; Sequence Analyses; Sequence Analysis; Simian Immunodeficiency Viruses; SIV; Staging; Surface; Techniques; Testing; Time; tool; Variant; Variation; Viral; Virus; Virus-HIV; Virus-Retrovirus; Viruses, General; Work
Relevance: Humans and laboratory animals have an array of antiviral defense mechanisms, of which we have only a rudimentary understanding. Identifying and understanding the mechanism of action of antiretroviral gene products could lead to completely new chemotherapeutic strategies for tackling infectious diseases, including AIDS. In addition, understanding species-specific variation and engineering resistance to antiretroviral genes could facilitate the development of animal models of human retroviral infection
Project start date: 2005-01-01
Project end date: 2016-01-31
Budget start date: 15-FEB-2011
Budget end date: 31-JAN-2012
PFA/PA: PA-10-067
2R01AI064003-06 (2011): $465000
2R37AI064003-06 (2011): $465000
Sponsored Links Excellgen http://Excellgen.com
NEW TECHNOLOGIES, NOVEL DISEASES; INDUSTRIAL ILLNESS IN 20TH CENTURY RAYON MANUFA
D Paul
University Of California San Franciscocity: San Francisco country: United States (us)
Grant 1G13LM010076-01A1 from National Library Of Medicine
Abstract: The aim of this proposed project is to complete a scholarly, book-length monograph, "New Technologies, Novel Diseases Industrial Illness in 20th Century Rayon Manufacturing." This will examine the manner in which a well-defined industry employing a unique manufacturing process was inextricably linked to a widespread outbreak of severe disease with continuing manifestations of toxicity that became endemic within its workforce. This industry spanned the entirety of the 20th century, continuing to cause industrial illnesses throughout this period. This case study poses the central question When a new technology leads to novel adverse health effects, what are the barriers to and promoting factors for effective preventive measures? This monograph, in six principal chapters, will integrate intellectual history (technological innovation and the medical recognition of a novel hazard), political-economic history (studying what was an early and even prototypical multinational enterprise), and social history (viscose being both a key war materiel and a mass-marketed consumer product). The analysis will approach carbon disulfide-caused occupational disease in the viscose industry as a case study, employing an historical narrative approach. Study materials will be derived from systematic survey of primary published medico-scientific materials (including journal articles, textbooks, and scientific meeting proceedings); primary regulatory and other governmental materials (including labor inspection data); other published materials (such industrial histories and biographical sources); archival and other resources (for example, material related to social historical aspects, such as artifacts of popular culture). The introduction of new technologies, the emergence of toxicity, its biomedical recognition, the dissemination of knowledge regarding the problem, and the responses of the industry, affected employees, and governmental agents, will all be considered. The target audience includes occupational and environmental health clinicians and public health practitioners; historians of medicine, technology, science studies, social studies, and economics; and policy makers and regulators. The proposed project is significant because it addresses the transnational history of an understudied topic in occupational health with important contemporary implications for disease prevention. The relevance of this project to public health arises from the ways in which learning the lessons of the past better informs our protective strategies going forward in time, particularly insofar as emerging threats from novel technologies are concerned
Keywords: Accounting; Address; Affect; Artifacts; base; Benzene; Benzol; Benzole; Books; Carbon Disulfide; case history; case report; Case Study; Cellophane; Chemicals; consumer product; Cyclohexatriene; Data; Disease; Disease Outbreaks; Disease Outcome; disease prevention; disease/disorder; Disorder; disorder prevention; driving force; Economics; Elements; Employee; Environmental Health; Environmental Health Science; experience; fabric; hazard; Health; Historical Aspects; History; History, Modern 1601-; Hygiene, Industrial; Industrial Health; Industrial Medicine; Industry; International; Investigation; journal article; Journal Article; Journal Article (PT); Journal Article [Publication Type]; Knowledge; language translation; Learning; Length; Link; manufacturing process; Marketing; Measures; Medical; Medicine; meetings; Modeling; Modern 1601-history; Modern History; Monograph; Monograph (PT); Monograph [Publication Type]; Morphologic artifacts; new technology; novel; occupational disease/disorder; Occupational Diseases; occupational disorder; Occupational Health; Occupational Medicine; Occupational or Industrial Medicine; Outbreaks; Play; poison; Poisons; Policy Maker; Polymer Chemistry; Prevention; Preventive; Public Health; public health medicine (field); public health relevance; Publishing; rayon; Recording of previous events; Research Resources; Resources; response; Role; Route; Science; Science of Medicine; social; social role; Source; Standardization; Survey Instrument; Surveys; technological innovation; Technology; Textbooks; Textiles; Time; Toxic Chemical; toxic compound; Toxic effect; Toxic Substance; toxicant; Toxicities; Translating; Translatings; War
Relevance: New Technologies, Novel Diseases: Industrial Illness in 20th Century Rayon Manufacturing Public Health Relevance The proposed project , a book-length monograph, addresses the central question: When a new technology leads to novel adverse health effects, what are the barriers to and promoting factors for effective preventive measures? The introduction of new technologies, the emergence of toxicity, its biomedical recognition, the dissemination of knowledge regarding the problem, and the responses of the industry, affected employees, and governmental agents, will all be considered. The proposed project is highly relevant to public health because it addresses the transnational history of an understudied topic in occupational health with important contemporary implications for disease prevention. Learning the lessons of the past better informs our public health protective strategies going forward in time, particularly insofar as emerging threats from novel technologies are concerned
Project start date: 2011-03-15
Project end date: 2014-03-14
Budget start date: 15-MAR-2011
Budget end date: 14-MAR-2012
PFA/PA: PAR-09-030
1G13LM010076-01A1 (2011): $50000
EFFECT OF ARGININE METABOLISM ON BIOFILM FORMATION IN THE STAPHYLOCCOCI
D Paul
University Of Nebraska Medical Centercity: Omaha country: United States (us)
Abstract: Staphylococci are a major burden on our society causing significant morbidity, mortality, and increased cost in healthcare. Increasing this disease burden is their ability to form biofilms on biomaterials resulfing in increased tolerance to anfibiofics and action of the immune system. Further study on the development and maturation of staphylococcal biofilms may lead to novel therapeufics that lead to their disruption. Staphylococcal biofilms are known to display spatial heterogeneity containing several physiological states including aerobic and anaerobic regions. Maintenance of these physiological states is imperative to the development of a mature biofilm. It is hypothesized that arginine catabolism is crucial for the development and maturation of anaerobic regions within both a Staphylococcus aureus and S. epidermidis biofilm. In fact, several backgrounds of both S. aureus and S. epidermidis contain two complete copies of the arginine deiminase (ADI) operon, one of which is acquired on a pathogenicity island. The ADI operon synthesizes proteins which catabolize arginine resulfing in ATP and ammonia. Within certain microniches of a biofllm environment, the resulting ATP can be used for metabolic purposes whereas the ammonia may be used for pH homeostasis. Studies demonstrate that arginine metabolism is induced during biofilm development and that the acquired (i.e. from pathogenicity island) ADI operon is most transcriptionally active as the biofilm matures. The importance of arginine metabolism will be studied by first exploring the regulatory role of ArcR, a known regulator of ADI expression, during biofilm growth. It is hypothesized that maximal inducfion of the acquired ADI operon occurs during biofilm growth and that Induction under these conditions is ArcR- dependent Secondly, through the use of fluorescent gene fusions, we will examine the temporal and spafial pattern of ADI gene expression within a S. epidermidis and S. aureus biofilm and compare those data to known anaerobic and aerobic regions of a biofilm. Lasfiy, through a mouse foreign body infection model, the relative virulence of ADI mutants in comparison to wild type S. aureus and S. epidermidis w´ be determined. Suscepfibility of ADI mutants to anfibiotics will be tested in a guinea pig fissue cage model. RELEVANCE (See instructions)
Keywords: Address; Adherence (attribute); Aerobic; Ammonia; Antibiotic Therapy; Arginine; Arginine deiminase; Bacteremia; base; Biocompatible Materials; burden of illness; Catabolism; Cavia; Cells; cost; Data; design; Development; Disease; DNA; Environment; extracellular; Foreign Bodies; Gene Cluster; Gene Expression; Gene Fusion; Genes; Genetic; Genus staphylococcus; Glucose; Growth; Healthcare; Heterogeneity; Immune system; Infection; Instruction; Island; Lead; Link; Maintenance; Metabolic; Metabolism; Microbial Biofilms; Minor; Modeling; Morbidity - disease rate; Mortality Vital Statistics; Mus; mutant; Nitric Oxide; novel; Operon; Oryctolagus cuniculus; Pathogenicity Island; Pathway interactions; Pattern; pH Homeostasis; Physiological; polysaccharide intercellular adhesin; Predisposition; Proteins; Regulation; Relative (related person); Reporter Genes; research study; Role; Societies; Staphylococcus aureus; Staphylococcus epidermidis; Testing; Transcriptional Regulation; Virulence
Budget start date: 1-JUL-2011
Budget end date: 30-JUN-2012
5P01AI083211-03_5328 (2011): $306888
PROTEIN MASS SPECTROMETRY (SHARED RESOURCE)
D Paul, Professor
University Of California Irvinecity: Irvine country: United States (us)
Abstract: Protein identification and proteomics are important new tools in cancer research. The Protein Mass Spectrometry Shared Resource will provide high-level mass spectrometry of biomolecules and protein analyses. This Shared Resource is based around a pair of tandem mass analyzers (the AB 4700 MALDITOF/ TOF and Thermo LTQ nanoESI-QIT), with access to as many as ten HPLC systems containing 10 quaternary and two binary pumps, seven PCs running at least 21 software packages, and numerous smaller items. The Facility is able to routinely identify proteins in slices from polyacryamide gels. In addition, it can routinely identify proteins in pull-down mixtures, complex virions, and whole cell extracts. They have done multi-dimensional (up to three dimensions) of tandem HPLC prior to LC-MALDI analysis. This capability includes two-dimensional protein fractionation (PF2D) and two and three dimensions of tandem peptide fractionation. This Facility provides an exciting springboard to the frontiers of state-of-the art proteomics. The Protein Mass Spectrometry Facility has been supported by the Cancer Center as a developing Shared Fesource for the past three years. Permanent funds are now requested for support as a full Shared Resource
Keywords: 2-dimensional; Amides; anticancer research; Assay; base; Binding Sites; Bioassay; Biologic Assays; Biological Assay; biomarker; Cancer Center; Cancer Center of University of California Irvine; Cancer Center Support Grant; cancer research; CCSG; Cell Extracts; Chao Family Comprehensive Cancer Center of University of California Irvine; Chemical Fractionation; Chromatography, High Performance Liquid; Chromatography, High Pressure Liquid; Chromatography, High Speed Liquid; Combining Site; Complex Mixtures; computer program/software; Computer Programs; Computer software; Core Grant; Data; Deuterium; Development; Dimensions; FRACN; Fractionation; Fractionation Radiotherapy; frontier; Funding; Future; Gel; gene product; H element; H+ element; H2 isotope; High Pressure Liquid Chromatography; HPLC; Hydrogen; Hydrogen Ions; in vivo; Individual; instrument; Ions; Isotopes; Label; Ligand Binding; MALD-MS; MALDI; MALDI-MS; mass analyzer; mass spectrometer; mass spectrometry shared resource; Mass Spectrum; Mass Spectrum Analysis; matrix assisted laser desorption ionization; Measures; Methods; Methods and Techniques; Methods, Other; nano-electrospray; nanoelectrospray; P30 Grant; Peptides; Phosphopeptides; Phosphorylated Peptide; Photometry/Spectrum Analysis, Mass; Property; Property, LOINC Axis 2; protein complex; Proteins; Proteomics; Protocol; Protocols documentation; Protons; Pump; Reactive Site; Reagent; Relative; Relative (related person); Resolution; Resource Sharing; Robotics; Running; Sampling; Scheme; Services; Shotguns; Slice; Software; Solvents; Spectrometry, Mass; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; Spectroscopy, Mass; Spectroscopy, Mass, Matrix-Assisted Laser Desorption-Ionization; Spectrum Analyses, Mass; Spectrum Analysis, Mass; Spottings; System; System, LOINC Axis 4; Techniques; tool; two-dimensional; University of California Irvine Cancer Center; Virion; Virus Particle; Work
Budget start date: 1-FEB-2011
Budget end date: 31-JAN-2012
5P30CA062203-16_5615 (2011): $20554
SEQUENCED VS. INTEGRATED DELIVERY OF TREATMENT FOR ADOLESCENT DEPRESSION AND SUD
D Paul, Senior Scientist
Oregon Research Institutecity: Eugene country: United States (us)
Grant 5R01DA021357-05 from National Institute On Drug Abuse
Abstract: Project Summary. Although the frequency of comorbidity is a well-established aspect of adolescent psychopathology, little is known regarding the effective delivery of treatment services to youth with comorbid conditions. In an effort to improve treatment engagement, response, and maintenance of gains, the proposed study will evaluate service delivery methods of integrating two empirically-supported interventions, the Adolescent Coping With Depression course (ACWD) and Functional Family Therapy (FFT), for depressive disorder and substance use disorders (SLID), respectively. Over a 5-year period, 180 adolescents (ages 13-17) from Portland, OR and Albuquerque, NM diagnosed with depression and SUD, and their parents/guardians will be recruited and randomly assigned to one of three treatment delivery conditions (a) FFT followed by ACWD, (b) ACWD followed by FFT, or (c) an integrated intervention combining and augmenting the ACWD and FFT interventions. Treatment in each condition will consist of 24 (12 families and 12 groups) sessions provided over a 20 week period. Participants will be assessed at intake, after 6,12, 18, and 24 treatment sessions (i.e., Post-Treatment), and at 3-, 6-, and 9-month follow-up. By providing ACWD and FFT interventions in both sequences, we will evaluate whether depression maintains SUD (Self-Medication model), SUD maintains depression (Affective Consequences model), each disorder maintains the other (Reciprocal Relations model), or both are maintained by other variables (Independent Factors model). In addition, integrating interventions to target both disorders in a coordinated fashion is expected to enhance the efficacy of treatment for youth with co-occurring disorders compared to a sequence of treatment that targets only one disorder at a time. Moreover, because the integrated treatment will address both conditions in early sessions, higher engagement rates are expected, compared to either sequenced treatment delivery. The study will also examine whether depressotypic cognitions and pleasant activities mediate the effects of ACWD and if improved family functioning mediates the effects of FFT. Relative severity or temporal onset of the two disorders will be examined as potential moderators of treatment outcome. Relevance. The co-occurrence of depression and SUD exacerbates problems in functioning and increases distress (e.g., elevated rates of suicide attempts and completion). Comorbidity also complicates the selection and delivery of treatment services and is generally associated with higher drop-out and lower treatment response. The proposed project represents the first study to experimentally evaluate interventions for adolescents with co-occurring depression and substance use disorders. Testing the combination and sequencing of empirically supported treatments will provide critical information regarding the influence of each disorder on the other and yield important information regarding how to most effectively provide treatment services in a clinically meaningful and data-driven manner
Keywords: Acute; Address; Adolescent; Affective; Aftercare; Age; Alcohol or Other Drugs use; arm; Attention; base; Clinical Trials; Cognition; Cognitive Therapy; cohesion; Comorbidity; Conduct Disorder; Conflict (Psychology); control trial; coping; Data; Depressed mood; Depressive disorder; Diagnosis; Disease; Distress; Drops; Dysthymic Disorder; effective therapy; Family; Family psychotherapy; follow-up; Frequencies (time pattern); Functional disorder; improved; Individual; Intake; Intervention; Maintenance; Major Depressive Disorder; Measures; Mediating; Mediator of activation protein; meetings; Mental Depression; Methods; Modeling; Mood Disorders; New Mexico; Oregon; Outcome; Outcome Measure; parental monitoring; Parents; Participant; Phase; primary outcome; Process; Psychopathology; Randomized; Randomized Controlled Trials; Recovery; Recruitment Activity; Relapse; Relative (related person); Research; response; Sampling; Self Medication; Services; Severities; Staging; Substance Use Disorder; substance using adolescents; Suicide attempt; Symptoms; System; Testing; Time; treatment effect; Treatment Efficacy; Treatment outcome; treatment response; Youth
Project start date: 2007-09-30
Project end date: 2012-06-30
Budget start date: 1-JUL-2011
Budget end date: 30-JUN-2012
PFA/PA: PA-05-139
5R01DA021357-05 (2011): $517931
PERIPHERAL BIOMARKERS IN FAMILIAL ALZHEIMER´S DISEASE
D Paul
Sun Health Research Institutecity: Sun City country: United States (us)
Grant 5R21AG030429-02 from National Institute On Aging
Abstract: Our previous studies (presented in Preliminary Results) have shown that multivariate analysis of transcripts related to inflammatory response and to cellular stress can distinguish Alzheimer´s disease peripheral blood leukocytes from control leukocytes. This ability to distinguish clinically diagnosed AD from controls on the basis of expression profile of easily obtained samples raises the question; can we distinguish preclinical AD on the basis of gene expression profiles of peripheral blood leukocytes? To answer this question requires obtaining blood samples from a cohort of non-demented persons then following them longitudinally for years to detect new AD cases in the cohort. The cohort may be enriched by selecting persons at risk, as was done in the ADAPT study which defined risk on the basis of age and of AD in a first degree relative. In the study proposed here we enrich our cohort of non-demented persons at risk for AD on the basis of carrying one of the well-defined familial gene mutations that lead, inevitably, to subsequent AD. In this proposal we focus largely on the mutated gene for PS1 since it is a frequently mutated gene among the FAD gene mutations. The central hypothesis to be tested by the research proposed here is, then; that those persons with an FAD mutation but diagnosis of no AD represent "preclinical" AD and that they will present a gene expression profile that will be predictive of subsequent AD by virtue of being similar to the AD gene expression profile of family members with the same mutation and with AD. We will extract RNA from peripheral blood leukocytes obtained from three classes of persons 1) with an FAD mutation and with diagnosed AD, 2) with an FAD mutation and diagnosis of no AD and 3) wild type with no FAD mutation and a diagnosis of no AD. Where possible, these three classes will be derived from the same family. Gene expression in the extracted RNA will be examined by quantitative RT-PCR (qRT-PCR) with the targets being those transcripts related to inflammatory response and cellular stress that we have defined in our earlier work. Statistical analysis of these data will be by means of multivariate discriminant analysis, as we have done in our earlier work. Additionally, we will use standard and newly developed statistical methods for the analysis of array data to be collected on these same samples with the aim of determining whether alternate transcripts within the classes of inflammatory and cell stress (as well as other classes) may be superior to those we have previously defined. Data from the study proposed here will be interpreted within the framework of parallel study of analysis of protein expression in these same FAD cases (by Timothy Mhyre), as well as within the framework of our almost completed (Quad) study of late onset AD (LOAD) using the same methods as proposed here. The Quad Study has four groups of ~20 each AD, MCI, PD and control. We have also initiated a study of gene expression in peripheral leukocytes in LOAD at Sun Health Research Institute where we have access to over 150,000 elderly. Alzheimer´s disease is a major public health problem in the United States, as well as world-wide. In the U.S. the number of cases is 4.5 - 5 million with an annual Medicare cost of about $100 billion. As the population of the U.S. ages, the number of Alzheimer´s cases is projected to grow to 16 million by 2050, with consequent immense increased expenses. The need for more effective and early diagnosis, coupled with more effective treatment that would slow or halt disease progression is clearly a pressing need. The study proposed here addresses the diagnosis arm of these needs. A number of studies have established that Alzheimer´s disease has been damaging the brain for decades before symptoms become evident enough to come to medical attention. Unfortunately, by that time a large percentage of nerve cells have been lost in critical areas of the brain concerned with memory, judgment and cognitive skills. This emphasizes the importance of detecting Alzheimer´s disease as early as possible so that treatment can be initiated before significant brain damage has taken place. We have obtained data that demonstrates the ability to distinguish already diagnosed Alzheimer´s disease from control cases on the basis of a profile of gene expression by easily obtained peripheral blood leukocytes. The question addressed by the proposal submitted here is whether our method which has been successful at distinguishing already diagnosed disease can be successful at predicting a clinical diagnosis of Alzheimer´s disease years later. In order to do this we propose to study families with gene mutations that cause early Alzheimer´s disease. Some family members with such a mutation will already have AD, other family members with the mutation will not yet have been clinically diagnosed with AD, while still other family members will have neither the mutation nor Alzheimer´s disease. Profiling gene expression in peripheral blood cells of these three classes of family members will tell us whether family members with the mutation but not yet showing clinically detected Alzheimer´s disease show a gene expression profile similar to that of family members with the mutation who have been diagnosed with AD. If this is so, it would constitute a finding that it is possible to detect Alzheimer´s disease prior to the appearance of clinically detectable symptoms in cases of familial AD, and will provide a basis for extension of these studies to the more common late onset Alzheimer´s disease
Keywords: Acquired brain injury; Address; advanced age; Age; Aged 65 and Over; Alzheimer; Alzheimer disease; Alzheimer sclerosis; Alzheimer syndrome; Alzheimer`s; Alzheimer`s Disease; Alzheimer`s disease risk; Alzheimers Dementia; Alzheimers disease; Amentia; analytical method; Appearance; Area; Arm; Arts; base; biomarker; Blood Cells; Blood leukocyte; Blood Sample; Blood specimen; Boils; Brain; brain damage; Brain Injuries; brain lesion (from injury); Caring; Cellular Stress; clinical Diagnosis; Clinical Management; Code; Coding System; Cognitive; cohort; cost; Coupled; Data; Data Analysis, Statistical; Data Interpretation, Statistical; Dementia; dementia of the Alzheimer type; Dementia, Alzheimer Type; Dementia, Primary Senile Degenerative; Dementia, Senile; density; design; designing; Detection; Diagnosis; Discriminant Analyses; Discriminant Analysis; Disease; disease control; disease diagnosis; Disease Progression; disease/disorder; Disorder; disorder control; DNA Alteration; DNA mutation; early detection; Early Diagnosis; early onset AD; Early Onset Alzheimer Disease; effective therapy; Elderly; Elderly, over 65; elders; Encephalon; Encephalons; EOAD; Evaluation; Exhibits; Expression Profiling; Expression Signature; familial AD; familial Alzheimer disease; familial Alzheimer disease (FAD); Family; Family member; Family Study; First Degree Relative; Furuncles; Gene Alteration; Gene Expression; Gene Expression Profile; gene expression signature; Gene Mutation; Gene Products, RNA; Genes; Genetic Alteration; Genetic Change; Genetic defect; Genetic mutation; genome mutation; geriatric; Hand; Health; Health Insurance for Aged and Disabled, Title 18; Health Insurance for Aged, Title 18; health insurance for disabled; Health Insurance for Disabled Title 18; Heating; heavy metal lead; heavy metal Pb; improved; Inflammatory; Inflammatory Response; Judgment; late life; Late Onset Alzheimer Disease; later life; Lead; Leukocytes; Maps; Marrow leukocyte; medical attention; Medicare; Memory; Methods; molecuar profile; Molecular Fingerprinting; Molecular Profiling; molecular signature; Multivariate Analyses; Multivariate Analysis; Mutate; Mutation; Nerve Cells; Nerve Unit; Nervous System, Brain; Neural Cell; Neurocyte; neuronal; Neurons; older adult; older person; Pb element; Peripheral; peripheral blood; Peripheral Blood Cell; Persons; Population; Position; Positioning Attribute; pre-clinical; preclinical; Presenile Alzheimer Dementia; primary degenerative dementia; Primary Senile Degenerative Dementia; Probability; Procedures; protein expression; Public Health; public health medicine (field); public health relevance; Research; Research Institute; Reticuloendothelial System, Leukocytes; reverse transcriptase PCR; Reverse Transcriptase Polymerase Chain Reaction; Ribonucleic Acid; Risk; RNA; RNA, Non-Polyadenylated; Robotics; RT-PCR; RTPCR; sample collection; Sampling; senile dementia of the Alzheimer type; senior citizen; Sequence Alteration; skills; specimen collection; Statistical Data Analyses; Statistical Data Interpretation; Statistical Methods; Symptoms; Testing; The Sun; Time; Title 18; Transcript; transcriptome; United States; Universities; Upper arm; virtual; white blood cell; White Blood Cells; white blood corpuscle; White Cell; Work
Project start date: 2008-09-01
Project end date: 2011-08-31
Budget start date: 1-SEP-2009
Budget end date: 31-AUG-2011
PFA/PA: PA-06-181
5R21AG030429-02 (2009): $183813
UNCOVERING THE MOLECULAR BASIS OF MALIGNANT HYPERTHERMIA
D Paul, Professor
Brigham And Women´s Hospitalcity: Boston country: United States (us)
Grant 5P01AR052354-05 from National Institute Of Arthritis And Musculoskeletal And Skin Diseases
Abstract: Our overall hypothesis is that the mutations in RyR1 responsible for human Malignant Hyperthermia (MH) and Central Core Disease (CCD) alter the stability of the EC coupling macromolecular complex and the dynamics of Ca2+ signaling, These changes are responsible for susceptibility to agents that trigger MH and to create the weakness associated with CCD. Data from our dysgenic (mdg) and dyspedic mouse models have defined the critical regions of the slow voltage gated Ca2+ channel in the surface membrane (y is-DHPR) and RyR1 of the sarcoplasmic reticulum (SR) that are responsible for bi-directional signaling between these two proteins. We have also recently demonstrated a three-way interaction between RyR1, the vis-DHPR and a Ca2+-entry channel in the surface membrane. In vitro studies of RyRs and Y is-DHPRs with MH mutations expressed in null cell lines have demonstrated that they have a phenotype similar to muscle from MH susceptible humans and pigs. Three mouse models have been created expressing RyR1 with MH/CCD nutations. Additional mice will be created with other RyR1 mutations to allow us to study the relationship between severity of the phenotype and the site of a mutation. The themes of Projects 1-4 are closely interrelated, and the expertise from each of the project PI´s is unique and essential to the interdisciplinary goals of the proposed program. The administrative, tissue culture transgenic animal, human tissue and morphology cores will provide a unifying resource for common reagents and morphology studies for the four project investigators. The scope of investigations will range from creation and extensive phenotyping of MH and CCD mice (Projects 1 and 3), studies of protein function and interactions in isolated muscle triads (Project 2), influence of mutations on the RyR1 structure and biochemistry (Project 3), and their influence on the cellular physiology of muscle and dendritic cells (Projects 1, 2, & 4). The expected outcome of the work proposed by this Program Project is a better understanding how MH and CCD mutations of RyR1 differentially perturb the dynamics of cytoplasmic Ca2+ at rest and during activity, how they promote subtle to catastrophic cellular dysfunctions and their relationship to genetic background
Project start date: 2006-04-14
Project end date: 2012-03-31
Budget start date: 1-APR-2010
Budget end date: 31-MAR-2012
5P01AR052354-05 (2010): $1322554
D Paul, Professor
Brigham And Women´s Hospitalcity: Boston country: United States (us)
Abstract: It is the central tenant of Core B and this program project that the most appropriate method to study MH is in murine models that expresses frequently seen human MH mutations. Mouse models can provide sufficient tissues for molecular, biochemical, cellular, and physiological analyses by a multidiscipiinary team whose collective goal is to understand the etiology of these myopathies. The use of mice also permits these analyses to be performed in diverse genetic backgrounds. Core B will perform the repetitive tasks that are necessary to support all four Projects. This core will provide an important and integrated facility to receive gene-targeting constructs from Project 1 and Project 3. They will transfect these constructs into ES cells and after the projects identify homologously targeted clones, amplify these clones for blastocyst injection and inject these targeted ES cells into blastocysts to produce chimeras. After confirmation of germline transmission by Projects 1 and 3, Core B will produce MH "knock-in" mice to be studied by all four Projects. Core B will take cDNA constructs from Projects 1, 3 and 4 and package them into HSV1 (RyRs) or Lentivirus (DHPRs) virions to be used for expression of mutated proteins and intracellular reporters in myotube cultures and distribute them as needed to all 4 projects. Core B will make myoblast cell lines from all heterozygous and homozygous MH "knock-in" mice and maintain dyspedic, dysgenic and dyspedic/dysgenic cell lines to be used by all 4 projects. Core B will receive human MHS muscle samples from Core C and maintain myoblast cell lines from these samples to be used by projects 1 and 4. In addition to maintaining MH "knock-in" animal lines Core B will take responsibility for interbreeding these animals with C57BL6 and BalbC mice to produce MH animals with different genetic backgrounds The uniform and consistent supply of exactly the same study models to all four Projects by this Core will allow a truly integrated approach to the study of Malignant Hyperthermia
Keywords: anesthesia related hyperthermia; Animals; Biochemical; Blastocyst; blastocyst; Blastocyst structure; Blastosphere; blastula; Body Tissues; Causality; cDNA; Cell Line; Cell Lines, Strains; CellLine; Chimera; Chimera organism; Complementary DNA; cultured cell line; disease causation; disease etiology; disease/disorder etiology; disorder etiology; Disorder of muscle, unspecified; DNA, Complementary; Embryo, Preimplantation; embryonic stem cell; ES cell; Etiology; gene product; Gene Targeting; Genetic; Genetic Alteration; Genetic Change; Genetic defect; genome mutation; Goals; herpes simplex i; Herpes Simplex Virus 1; Herpes Simplex Virus Type 1; herpes virus 1, human; Herpesvirus 1 (alpha), Human; Herpesvirus 1, Human; HHV-1; HSV-1; HSV1; Human; human alphaherpesvirus 1; Human herpes simplex virus type 1; Human herpesvirus 1; Human herpesvirus type 1; Human, General; hyperthermia of anesthesia; Injection of therapeutic agent; Injections; Knock-in; Knock-in Mouse; Lentivirinae; Lentivirus; Malignant Hyperpyrexia; Malignant hyperpyrexia due to anesthesia; Malignant Hyperpyrexias; Malignant Hyperthermia; Mammals, Mice; Man (Taxonomy); Man, Modern; Methods; Mice; Modeling; Molecular; mouse model; Murine; Mus; Muscle; Muscle Cells, Embryonic; Muscle Cells, Precursor; Muscle Disease; Muscle disease or syndrome; Muscle Disorders; Muscle Fibers; Muscle Tissue; Muscular Diseases; muscular disorder; Mutate; Mutation; Myoblasts; Myopathic Conditions; Myopathic disease or syndrome; Myopathic Diseases and Syndromes; Myopathy; Myopathy, unspecified; Myotubes; Physiologic; Physiological; programs; Programs (PT); Programs [Publication Type]; Proteins; Reporter; Rhabdomyocyte; Sampling; Skeletal Fiber; Skeletal Muscle Cell; Skeletal Muscle Fiber; Skeletal Myocytes; stem cell of embryonic origin; Study models; Subfamily lentivirinae; Targetings, Gene; Tissues; Transmission; transmission process; Virion; Virus Particle; Virus-Lenti
Budget start date: 1-APR-2010
Budget end date: 31-MAR-2011
5P01AR052354-05_9001 (2010): $209624
LATE EVENTS IN RETROVIRUS ASSEMBLY
D Paul, Professor
Aaron Diamond Aids Research Centercity: New York country: United States (us)
Grant 5R01AI052774-10 from National Institute Of Allergy And Infectious Diseases
Abstract: In this competing renewal application we request support for our studies on late events in retrovirus assembly. In the previous funding cycle, we identified and characterized several cellular proteins that are recruited to sites of retrovirus assembly by viral late budding (L)-domains. These proteins are either (i) components of the class E vacuolar protein sorting (VPS) pathway that is normally involved in the manipulation of endosomes and multivesicular bodies (MVB) or (ii) ubiquitin ligases. For several of these factors we were able to demonstrate a critical role in the release of infectious retroviral particles. Several mechanistic questions remain, however, and recently we have identified additional cellular factors that seem very likely to be involved in the late stages of the construction and release of retrovirus particles, and/or endosome/MVB biogenesis. In the next grant period, we will attempt to elucidate how previously defined and newly identified cellular factors participate in the late stages of retrovirus assembly and related cellular functions by pursuing two specific aims. Specific Aim 1 is to understand how specific endosomal proteins are incorporated into HIV-1 particles and their role in generating infectious virions. Specific aim 2 is to determine the role of ubiquitin, ubiquitin ligases and novel ubiquitin ligase binding proteins, discovered during the previous funding period, in retroviral budding and trafficking of endocytosed proteins. Execution of these two aims should result in significant progress toward our long term goal of understanding how cellular and viral factors cooperate in the construction of complete, infectious retroviral particles. Understanding these events could potentially provide new opportunities to interfere in these processes for the treatment of diseases caused by HIV and other enveloped viruses. Many enveloped viruses, for example HIV-1, are responsible for life threatening infections. To spread from cell to cell they make use of cellular proteins to facilitate the release of accurately assembled virus particles. Understanding how this system works could give opportunities to intervene with targeted therapeutics
Keywords: Binding Proteins; Biogenesis; Cell physiology; Cells; Clathrin; Disease; Endocytosis; Endosomes; Event; Funding; Goals; Grant; HIV; HIV-1; Infection; Life; Mediating; Multivesicular Body; novel; particle; Pathway interactions; Process; Proteins; public health relevance; Recruitment Activity; Request for Applications; Retroviridae; Role; Site; Sorting - Cell Movement; Staging; System; therapeutic target; trafficking; Ubiquitin; ubiquitin ligase; Vacuolar Protein Sorting; Viral; Virion; Virus; Work
Project start date: 2002-05-15
Project end date: 2012-05-31
Budget start date: 1-JUN-2011
Budget end date: 31-MAY-2012
5R01AI052774-10 (2011): $332305
CENTER FOR INTERDISCIPLINARY RESEARCH ON AIDS
D Paul, Professor & Dean Of Public Health
Yale Universitycity: New Haven country: United States (us)
Grant 5P30MH062294-09 from National Institute Of Mental Health
Abstract: In 1997, the Center for Interdisciplinary Research on AIDS (CIRA) received a P01 Program Project grant from NIMH and NIDA to conduct research aimed at the prevention of HIV infection and reduction of the negative consequences of HIV disease in vulnerable and underserved populations. We successfully competed for a five-year P30 CSPAR grant in September 2001. With this application, we seek to renew this CSPAR grant for an additional 5 years of support. To date, a total of 86 externally funded research and training grants (69 research, 17 training) have been affiliated with the Center; 41 (32 research) of these are currently ongoing. Five additional research grants are currently under review. A total of 41 scientists have served as Principal Investigators on these different projects. An additional 21 have been awarded funds through our pilot project program during this same period. CIRA affiliated research will continue to identify the determinants of risky behaviors in vulnerable populations, develop and assess interventions to address this risk, analyze policy, laws, and structures associated with risk and prevention, and examine the global dimensions of the pandemic. In addition, we will support the conduct of translational research and collaborative research focusing on HIV prevention, care, and treatment in national and international settings. Wherever possible, this research will be interdisciplinary, drawing from the full range of theoretical and methodological expertise of CIRA scientists, who together, represent 22 different disciplines. Six Cores will support the conduct of research in these areas. An Administrative (Admin) Core will serve as the planning and decision-making body and will provide overall scientific leadership, and administrative management. A Development (Dev) Core will provide mechanisms for promoting and supporting new research and bringing new scientists, at all stages in their careers, to HIV/AIDS research. An Interdisciplinary Research Methods (IRM) Core will provide consultations, hands-on assistance, resources, and training in quantitative methods and biostatistics, qualitative methods and ethnography, cost effectiveness and mathematical modeling, and biological measures and analysis. The Community Research (CR) Core will facilitate community-based and translational research by CIRA scientists, build research capacity of community partners, and facilitate the dissemination of research findings. A Clinical and Health Services Research (CHSR) Core will foster the development of research focused on developing and testing clinical strategies to address HIV prevention, care, and treatment for patients with comorbid psychiatric, medical, substance use and behavioral conditions in domestic and international settings. The Law, Policy, and Ethics (LPE) Core will facilitate the development of new and support ongoing research with a legal, policy, ethics, or structural focus. It will also ensure that CIRA research meets the highest ethical and legal standards and that it is accessible to policymakers
Project start date: 2001-09-30
Project end date: 2013-06-30
Budget start date: 1-JUL-2011
Budget end date: 30-JUN-2012
PFA/PA: PAR-08-009
5P30MH062294-09 (2011): $2090970
MUSCLE: EXCITATION/CONTRACTION COUPLING GORDON RESEARCH CONFERENCE
D Paul, Professor
Gordon Research Conferencescity: West Kingston country: United States (us)
Grant 1R13AR062438-01 from National Institute Of Arthritis And Musculoskeletal And Skin Diseases
Abstract: Gordon Research Conference on Muscle Excitation-Contraction Coupling This is a proposal for partial support of the Gordon Research Conference (GRC) on Excitation-Contraction Coupling (ECC) in skeletal muscle, the only national or international meeting with this specific focus. The 2012 conference will be held in Europe at the GRC site in Les Diablerets, Switzerland. Calcium signaling figures importantly in the adaptive responses of healthy muscle to exercise and age mutations in a number central players in the ECC process cause muscle diseases such as 1. hypokalemic periodic paralysis (skeletal L-type Ca2+ channel (DHPR)) 2. malignant hyperthermia (RyR1, skeletal DHPR) 3. central core disease, multi-mini core disease and centro-nuclear myopathy (RyR1). 4. Brody´s disease (SERCA1). 5. Timothy syndrome (Cardiac DHPR). and 5. catecholaminergic polymorphic ventricular tachycardia (RyR2 and Calsequestrin 2). Key to understanding the mechanisms causing these diseases is understanding the affected protein´s normal function. In addition to muscle diseases that result from mutations to the proteins that participate in ECC, there are many diseases that result from mutations to proteins that are not directly involved in this process. However, because a disturbance in Ca2+ signaling is part of a pathological cascade that causes muscle damage there is substantial benefit to having these diseases discussed in the ECC Gordon conference. in response to exercise and aging and both are important to human health. In the time since the 2009 ECC Gordon conference, there has been important progress in our understanding of excitation-contraction coupling and other types of calcium signaling in muscle, as well as of the role of calcium in muscle disease, adaptation and repair. It is this progress that provides the basis for our planning of the 2012 Gordon ECC Gordon Conference. We feel confident that the meeting we have planned will stimulate participants to pursue new approaches and research directions in the fields of muscle ECC, calcium signaling and disease. The goals of the 2012 Muscle Excitation Contraction Coupling Gordon Research Conference are to discuss and evaluate critically new results related to the basic mechanisms that allow muscle to contract, and how these are affected by fatigue, age and mutations that cause muscle disease. This triennial conference is the only conference devoted to this very important area of medically relevant research and will allow participants who do basic, translational and clinical research to meet and discuss new ways to solve important problems affecting everyone´s everyday life
Keywords: Affect; Age; Aging; Area; base; Basic Science; Calcium; Calcium Signaling; Calsequestrin; Cardiac; Cells; Central Core Myopathy; Clinical Research; college; Contracts; Coupling; Development; Disease; Disease Progression; disease-causing mutation; Environment; Europe; Exercise; experience; Fatigue; Goals; Health; Human; human disease; Hypokalemic periodic paralysis; International; Knowledge; Life; Malignant hyperpyrexia due to anesthesia; Measures; meetings; Methods; Molecular; Muscle; Mutation; Myopathy; novel strategies; Nuclear; Participant; Process; Proteins; repaired; Research; Research Personnel; response; Role; RyR1; RyR2; Scientist; SERCA1; Signal Transduction; Site; skeletal; Skeletal muscle structure; Switzerland; symposium; Time; Timothy syndrome; Translational Research; Triad Acrylic Resin; Ventricular Tachycardia
Relevance: The goals of the 2012 Muscle: Excitation Contraction Coupling Gordon Research Conference are to discuss and evaluate critically new results related to the basic mechanisms that allow muscle to contract, and how these are affected by fatigue, age and mutations that cause muscle disease. This triennial conference is the only conference devoted to this very important area of medically relevant research and will allow participants who do basic, translational and clinical research to meet and discuss new ways to solve important problems affecting everyone´s everyday life
Project start date: 2011-09-20
Project end date: 2012-08-31
Budget start date: 20-SEP-2011
Budget end date: 31-AUG-2012
PFA/PA: PA-10-071
1R13AR062438-01 (2011): $22000
Sponsored Links Excellgen http://Excellgen.com
PHOSPHORYLATION OF GAP JUNCTION PROTEINS
D Paul, Member
Fred Hutchinson Cancer Research Centercity: Seattle country: United States (us)
Grant 5R01GM055632-14 from National Institute Of General Medical Sciences
Abstract: Gap junctions are specialized matched membrane domains that contain channels that allow exchange of small molecules including ions, metabolites, and second messengers (e.g., Ca2+ and IP3) between neighboring cells. These channels are necessary for proper development and genetic linkage analyses have implicated connexins in at least 14 human diseases. The gap junction protein connexin43 (Cx43) is regulated via phosphorylation and its interactions with other proteins. This proposal focuses on the role that these two regulatory processes play and interplay in vivo to affect tissue development and function. We will examine the role Cx43 regulation plays during fundamental biological processes such as in the heart during ischemia, tachycardia and preconditioning, in skin during wound repair, and in the eye during development. We propose to (1) determine the consequences of specific Cx43 phosphorylation events on Cx43 function. (2) characterize changes in Cx43 phosphorylation and function in skin and heart in response to conditions such as wounding and hypoxia/ischemia., and (3) investigate the in vivo role of Cx43 phosphorylation in skin, heart, ovary, and eye at different developmental stages using "knock-in" mice expressing phosphorylation site mutants of Cx43. Understanding the linkage of changes in Cx43 phosphorylation to the exquisite control of fundamental biological events is in itself important but given that drugs to affect Cx43-related cardiac function and Cx43 anti-sense gels to speed wound healing are being tested in humans, we need to fully define these medically important biological events to better understand their implications and opportunities for patient treatment. We propose to investigate the linkage of changes in the phosphorylation of the gap junction protein connexin43 and gap junctional communication to the exquisite control of cellular proliferation and migration during development, cardiac stress, and wound healing. Since Cx43 anti-sense treatments for wound healing and drugs to affect Cx43-related cardiac function are currently being tested in humans, our results will help us to better understand current drug implications for patient treatment and may lead to better alternatives
Keywords: 21+ years old; Activation Analysis; Adult; adult human (21+); Affect; Affinity; Amino Acid Substitution; Amino Acids; aminoacid; Analyses, Activation; Antibodies; Assay; Bioassay; Biologic Assays; Biological; Biological Assay; Biological Function; Biological Process; biological signal transduction; Body Tissues; Brain Hypoxia-Ischemia; C-terminal; Calcium Phospholipid-Dependent Protein Kinase; Calcium-Activated Phospholipid-Dependent Kinase; Cardiac; cardiac muscle; Cell Communication and Signaling; Cell Cycle; Cell Cycle Stage; Cell Division Cycle; Cell Function; Cell Growth in Number; Cell Line; Cell Lines, Strains; Cell Locomotion; Cell Migration; cell motility; Cell Movement; Cell Multiplication; Cell physiology; Cell Process; Cell Proliferation; Cell Signaling; cell type; CellLine; Cells; Cellular Function; Cellular Migration; Cellular Physiology; Cellular Process; Cellular Proliferation; Communicating Junction; Communication; Complex; conformation; conformational state; Connexin 43; Connexin43; Connexins; cultured cell line; Culturing, in vitro Vertebrate, Primary; Cx43; Development; drug/agent; Drugs; EC 2.7; Event; Extracellular Signal-Regulated Kinase Gene; Eye; Eye Development; eye morphogenesis; Eyeball; family based linkage study; gap junction channel; Gap Junction Proteins; Gap Junctions; gatekeeper; Gatekeeping; Gel; gene product; Generalized Growth; Genetic Alteration; Genetic Change; Genetic defect; genetic linkage analyses; genetic linkage analysis; Genital System, Female, Ovary; genome mutation; GFAC; Grant; Growth; Growth Agents; Growth Factor; Growth Factors, Proteins; Growth Substances; Heart; heart muscle; heavy metal lead; heavy metal Pb; Human; human disease; Human, Adult; Human, General; Hypoxia; hypoxia ischemia; Hypoxia-Ischemia, Brain; Hypoxic; in vivo; injury response; Intracellular Communication and Signaling; Intracellular Second Messengers; Ions; Ischemia; Kinases; Knock-in; Knock-in Mouse; L-Serine; Lead; life course; Life Cycle; Life Cycle Stages; Link; linkage analyses; Linkage Analysis; Low-resistance Junction; Man (Taxonomy); Man, Modern; MAP Kinase Gene; MAPK; Measures; Medication; Membrane; membrane structure; migration; Mitogen-Activated Protein Kinase Gene; Molecular Configuration; Molecular Conformation; Molecular Stereochemistry; Motility; Motility, Cellular; mRNA Expression; Muscle, Cardiac; Muscle, Heart; mutant; Mutation; Myocardium; Nexus; Nexus Junction; ocular development; ontogeny; Output; Ovary; Oxygen Deficiency; Patients; Pb element; Pharmaceutic Preparations; Pharmaceutical Preparations; Phospholipid-Sensitive Calcium-Dependent Protein Kinase; Phosphorylation; Phosphorylation Site; Phosphotransferases; PKC; Play; preconditioning; prevent; preventing; Primary Cell Cultures; Process; Protein Kinase C; Protein Phosphorylation; Protein Trafficking; protein transport; Proteins; Proteomics; public health relevance; Reagent; Regulation; response; response to injury; Retina; Role; second messenger; Second Messenger Systems; Second Messengers; Serine; Signal Pathway; Signal Transduction; Signal Transduction Systems; Signaling; Site; Skin; small molecule; social role; spatiotemporal; Speed; Speed (motion); src Oncogenes; Staging; Stress; Structure; Subcellular Process; Tachycardia; Testing; Tissue Growth; tissue repair; Tissues; trafficking; Traffickings, Protein; Transphosphorylases; v src; v-src Genes; v-src Oncogenes; Vibrissae; Whiskers; Wound Healing; Wound Repair
Relevance: We propose to investigate the linkage of changes in the phosphorylation of the gap junction protein connexin43 and gap junctional communication to the exquisite control of cellular proliferation and migration during development, cardiac stress, and wound healing. Since Cx43 anti-sense treatments for wound healing and drugs to affect Cx43-related cardiac function are currently being tested in humans, our results will help us to better understand current drug implications for patient treatment and may lead to better alternatives
Project start date: 1997-05-01
Project end date: 2013-12-31
Budget start date: 1-JAN-2011
Budget end date: 31-DEC-2011
PFA/PA: PA-07-070
5R01GM055632-14 (2011): $338986
MOLECULAR AND GENETIC ANALYSIS OF CHROMATIN STRUCTURE
D Paul, Professor Of Molecular Biology
Princeton Universitycity: Princeton country: United States (us)
Grant 5R01GM043432-20 from National Institute Of General Medical Sciences
Abstract: Eukaryotes chromosomes are subdivided into functionally and structurally autonomous domains. While this was first suggested by cytological studies on the lampbrush chromosomes of amphibian oocytes and the polytene chromosomes of insects more than a half century ago, only little progress was made on this problem until 10-15 years ago. Since then a combination of genetic, molecular and biochemical experiments has provided convincing evidence for the domain organization of eukaryotic chromosomes. These studies have shown that the domain organization of the chromatin fiber is of critical importance not only for the packaging of chromosomes inside the nucleus, but also for the proper regulation of gene activity. The subdivision of eukaryotic chromosome into discrete domains requires a mechanism to separate one domain from another. Special elements called boundaries or insulators have been shown to serve this purpose. Elements that function as boundaries of chromatin domains were first identified in Drosophila, and they have now been found in a diverse array of organisms including from yeast to mammals. These elements delimit high order chromatin domains and function to establish independent units of gene activity, insulating genes or regulatory elements within a domain from the action of regulatory elements located outside in adjacent domains. The major aims of this proposal are to learn more about the biological functions of boundaries, to characterize the factors that confer insulator activity of several BX-C boundaries (including apparent stage and perhaps tissue specific factors) and to elucidate the mechanisms that enable boundaries to establish independent units of genetic activity. Narrative Boundary elements subdivided eukaryotes chromosomes into functionally and structurally autonomous domains. The major aims of this proposal are to learn more about the role of boundaries in development and elucidate the mechanisms that enable boundaries to establish independent units of genetic activity
Keywords: ing; Address; Amphibia; Biochemical; Biological Assay; Biological Process; Boundary Elements; Cell Nucleus; Chromatin; Chromatin Fiber; Chromatin Structure; Chromosomes; Complex; Development; Drosophila genus; Elements; Enhancers; Eukaryota; fly; Gene Expression Regulation; Genes; Genetic; genetic analysis; Goals; Health; Insecta; insight; Learning; Mammals; Modeling; Molecular Analysis; Molecular Genetics; novel; Oocytes; Organism; Process; Property; Regulation; Regulatory Element; research study; Role; Series; Staging; Testing; Tissues; Trans-Activators; Transgenes; Yeasts
Relevance: Narrative Boundary elements subdivided eukaryotes chromosomes into functionally and structurally autonomous domains. The major aims of this proposal are to learn more about the role of boundaries in development and elucidate the mechanisms that enable boundaries to establish independent units of genetic activity
Project start date: 1989-12-01
Project end date: 2013-07-31
Budget start date: 1-AUG-2011
Budget end date: 31-JUL-2012
PFA/PA: PA-07-070
5R01GM043432-20 (2011): $409847
THE FUNCTION OF ANTIMALARIAL DRUG RESISTANCE PROTEINS
D Paul
Georgetown Universitycity: Washington country: United States (us)
Grant 5R01AI056312-08 from National Institute Of Allergy And Infectious Diseases
Abstract: Drug resistant malaria kills millions annually. Reversing horrific trends in incidence and mortality requires a balanced approach in vaccine and drug research, as well as field based efforts to control vector populations and infection rates. Current and future treatment of the many different strains of drug resistant malaria that now exist requires a more complete understanding of multiple genotypes and phenotypes. We must not be lulled into a false sense of (temporary) security provided by current artemisinin (ART) based therapies; black market ART is already circulating and generating ART resistance. The struggle against drug resistant malaria is ongoing and must be met continuously; else we have learned nothing from the past 50 years while watching CQ and other drugs fail. We must "stay ahead of the resistance curve" and define molecular mechanisms that guides ongoing drug and vaccine research. Our laboratory has helped to lead the field in molecular level analysis of PfCRT and PfMDR1 proteins using heterologous expression systems. In this competitive renewal period we will Aim 1) Continue to define binding functions of PfCRT isoforms via heterlogous expression in yeast and analysis of purified membrane, ISOV, and PL preparations harboring these proteins. We will use recently developed techniques and chemical probes for drug, amino acid, and ion binding and transport. We will also synthesize additional probes (e.g.,AzB-MQ, AzBCQ side chain length variants, AzB-QN) for PfCRT function. These probes will also be used in Aim 3. Aim 2) Continue to define drug transport functions of PfCRT isoforms using ISOV and PLs and radio labeled and fluorescent (e.g. NBD CQ) probes. We will also synthesize additional probes (e.g., NBD-MQ, NBD-QN) using previously synthesized intermediates and similar chemistry relative to successful synthesis of NBD-CQ. Aim 3) Test hypotheses for function of PfMDR1 following a similar approach, and also using high throughput plate based ATPase assays we have developed and published [93, 93B]. We will investigate the unusual (relative to other ABCB transporters) drug - influenced "communication" between the two symmetrical halves of PfMDR1 [93]. We will analyze binding, transport and ATPase properties of ISOV and PLs harboring known ratios of various PfCRT and PfMDR1 proteins to test for interactions between the two transporters. Drug resistant malaria continues to both evolve and spread, and globally causes over 1 million deaths annually. This project aims to define, at a molecular level, how mutated proteins cause that drug resistance. Such information is central to development of new drugs and other therapies to combat drug resistant malaria
Keywords: Amino Acids; Anti-malarial drug resistance; artemisinine; Artemisinins; ATP phosphohydrolase; base; Binding (Molecular Function); Biological Assay; Cessation of life; Chemicals; Chemistry; combat; Communication; Development; Drug resistance; Drug Transport; Equilibrium; Future; Genotype; Incidence; Infection; Ions; Killings; Label; Laboratories; Lead; Learning; Length; Malaria; Marketing; meetings; Membrane; Molecular; Mortality Vital Statistics; Mutate; Pharmaceutical Preparations; Phenotype; Population; Preparation; Property; Protein Isoforms; Proteins; public health relevance; Publishing; Radio; Relative (related person); Research; Resistance; Security; Side; System; Techniques; Testing; trend; Vaccine Research; Vaccines; Variant; vector control; Yeasts
Project start date: 2003-06-15
Project end date: 2013-05-31
Budget start date: 1-JUN-2011
Budget end date: 31-MAY-2012
PFA/PA: PA-07-070
5R01AI056312-08 (2011): $338503
MAKING A QUANTUM LEAP IN PLAQUE RESEARCH WITH MODERN SCIENCES
D Paul, Professor
University Of California Los Angelescity: Los Angeles country: United States (us)
Grant 5R01DE020102-02 from National Institute Of Dental & Craniofacial Research
Abstract: Current tooth decay (dental caries) prevention methods include enamel hardening with fluoride and bacterial removal via mechanical and general antimicrobial approaches. These methods are based on the knowledge that oral plaque bacteria ferment dietary carbohydrates to produce pH-reducing organic acids. Initial reductions in caries incidence observed upon widespread implementation of these measures reached a plateaudecades ago. Today more that 40% of children under 10 years of age as well as more than 85% of the adult population in the United States still suffer from the disease that cause annual treatment cost of $ 80 billion. Further development of these "remove and kill all" approaches are not likely to significantly improve oral health. Revolutionary advancements can only be achieved by expanding our understanding of the microorganism- mediated processes leading to tooth decay. This will require a detailed picture of dental plaque organisms, their metabolic activities and interactions. The long-term objective of this application is to combine and apply (established) advanced technologies to provide a detailed understanding of the biological processes involved in cariogenesis. This will include a comprehensive analysis of the cariogenic potential of known pathogens and their influence on acid production. Furthermore, we will provide information on the metabolic activity of species whose function in acid production is currently unknown. In compliance with the mission of the NIDCR we will further develop targeted strategies against the current caries epidemic based on current knowledge and the new information developed with the advanced technologies in this project. We will combine sophisticated, species-specific in vivo labeling tools (monoclonal antibodies and fluorescent protein-expressing bacteria) with monitoring of acid production (pH-sensitive dyes, fluorescent proteins and NMR profiling), labeling of acid active species with stable isotope probing (SIP). These tools will reveal details of the processes in dental plaques regarding species, interspecies interactions and the metabolic processes contributing to cariogenic (acid-producing) or healthy (homeostatic) conditions. The second goal of this application will examine the potential of our previously developed specifically targeted antimicrobial peptides (STAMPs) against cariogenic Streptococcus mutans to shift plaque ecology towards a healthy plaque. We will further improve a current prototype antimicrobial peptide that is activated in acidic environments and develop more antimicrobial peptides against known cariogenic species as well as those identified in this project. This study will greatly expand our knowledge of the biological processes within plaques that lead to disease and provide novel therapeutic approaches that aim to achieve long-term oral health by specifically removing cariogenic species and leaving beneficial or harmless populations intact. Relevance to public health statement Tooth decay (caries) remains a major health issue in the United States and worldwide with a prevalence of more than 50% in young children that increases to about 85% in the adult population. The consequences of this disease range from a significant number missed days at school or work to malnutrition and effects on overall health, and result in about $80B in treatment costs. Caries disease-progression studies and resulting treatment regimen have not yielded significant oral-health improvements in several years. We propose to revisit the processes involved in caries development by combining carefully chosen and highly complementary new analytical and molecular biology tools. These tools will identify the roles of individual species and their characteristics involved in the acid production that leads to tooth decay. This new approach will provide a deeper understanding of tooth-decay progression and allow for the subsequent development of novel targeted therapeutic approaches to eliminate "bad" bacteria from the mouth for a long-term change to better oral health
Keywords: 10 year old; Acids; Address; Adult; antimicrobial; antimicrobial peptide; Bacteria; base; Behavior; Biological; Biological Process; Biology; Carbohydrates; Caries prevention; cariogenic bacteria; Characteristics; Chemistry; Child; Clinical Management; Collaborations; commensal microbes; Communities; Complex; Data; Dental; Dental caries; Dental Enamel; Dental Plaque; Dentistry; Detection; Development; Dietary Carbohydrates; Dimensions; Disease; Disease Progression; DNA; Dyes; Ecology; Effectiveness; Engineering; Environment; Epidemic; Excision; Fermentation; Fluorescent Dyes; Fluorescent Probes; Fluorides; Future; Gingiva; Goals; Green Fluorescent Proteins; Health; improved; In Situ; in vivo; Incidence; Incubated; Individual; insight; Institutes; interest; Isotopes; Killings; Knowledge; Label; Laboratories; Laser Scanning Confocal Microscopy; Lead; Learning; Left; Link; Malnutrition; Maps; Measures; Mechanics; Mediating; Metabolic; Metabolism; Methods; microbial; Microbial Biofilms; microbial community; microbicide; microorganism; Mission; Molecular Biology; Monitor; Monoclonal Antibodies; National Institute of Dental and Craniofacial Research; new therapeutic target; novel; novel strategies; novel therapeutic intervention; Oral; oral bacteria; oral biofilm; Oral cavity; Oral health; Oral Microbiology; oral pathogen; oral plaque; organic acid; Organism; Pacific Northwest; pathogen; Pathogenesis; Peptides; Population; Prevalence; Process; Production; Property; Proteins; prototype; public health medicine (field); public health relevance; quantum; Research; Research Personnel; research study; RNA; Role; Saliva; Sampling; Schools; Science; Source; Spatial Distribution; stable isotope; Stimulus; Streptococcus mutans; success; Sucrose; System; Techniques; Technology; Testing; Therapeutic Effect; Time; tool; Treatment Cost; Treatment Protocols; United States; Virulence; Work
Relevance: Relevance to public health statement Tooth decay (caries) remains a major health issue in the United States and worldwide with a prevalence of more than 50% in young children that increases to about 85% in the adult population. The consequences of this disease range from a significant number missed days at school or work to malnutrition and effects on overall health, and result in about $80B in treatment costs. Caries disease-progression studies and resulting treatment regimen have not yielded significant oral-health improvements in several years. We propose to revisit the processes involved in caries development by combining carefully chosen and highly complementary new analytical and molecular biology tools. These tools will identify the roles of individual species and their characteristics involved in the acid production that leads to tooth decay. This new approach will provide a deeper understanding of tooth-decay progression and allow for the subsequent development of novel targeted therapeutic approaches to eliminate "bad" bacteria from the mouth for a long-term change to better oral health
Project start date: 2010-05-01
Project end date: 2014-04-30
Budget start date: 1-MAY-2011
Budget end date: 30-APR-2012
PFA/PA: PA-07-070
5R01DE020102-02 (2011): $374287
REGULATION OF REPETITIVE CHROMATIN STRUCTURES DURING THE HUMAN CELL CYCLE
D Paul, Associate Professor
Univ Of Massachusetts Med Sch Worcestercity: Worcester country: United States (us)
Grant 2R01GM055712-14A1 from National Institute Of General Medical Sciences
Abstract: Chromatin Assembly Factor-1 (CAF-1) is a three-subunit protein complex conserved throughout eukaryotes. CAF-1 is a nucleosome assembly factor important for DNA replication, DNA repair, and heterochromatin formation. CAF-1 protein levels correlate with cell proliferation and cancer prognosis, making these studies a high priority for the medically important processes of genome stability and the maintenance of epigenetic states. Via mass spectrometry, we discovered multiple nucleolar proteins associated with the human CAF-1-p150 subunit. Microscopy detects a subset of cellular p150 associated with nucleoli, the sites of ribosomal RNA (rRNA) synthesis and ribosome assembly. Notably, RNAi-mediated depletion of p150 causes a dramatic loss of nascent rRNA transcripts and spatial redistribution of some nucleolar proteins. Therefore, we have discovered that p150 has a previously unrecognized role in the structure and function of the nucleolus. rRNA synthesis is regulated by energy supply, differentiation, cell cycle progression, tumor suppressors and oncoproteins. Nucleolar alterations are also important for cancer diagnoses, and the rRNA synthesis machinery is increasingly viewed as a therapeutic cancer target. Therefore, these studies are crucial for understanding clinically relevant interactions between DNA replication, gene expression, and growth control. We plan to explore three Aims to explore how p150 functions to regulate rRNA synthesis, and to extend these findings via genome-scale studies Aim 1. The mechanism of regulation of rDNA transcription by CAF-1 p150. We will test several hypotheses raised by our observations (a) p150 could be acting as part of a transcriptional activation complex at the rDNA promoter, perhaps distinct from its role as a CAF-1 subunit, (b) p150 could regulate the percentage of transcriptionally accessible rDNA repeats, (c) p150 could promote transcriptional elongation, or (d) p150 might be critical for maintaining the epigenetic modification state of the rDNA repeats. Alternatively, (e) p150 might prevent cryptic transcription events. We note that these possibilities are not mutually exclusive. Aim 2. Molecular analyses of p150 recruitment to repetitive DNAs. We will determine whether specific p150 protein domains are required for association with rDNA, and also assess the role of the new nucleolar interaction proteins in p150 recruitment. We will also determine the extent of cell cycle regulation of these associations. Aim 3. Genome-wide analysis of p150 localization, transcriptional targets, and effects on rDNA conformation. We will test our hypothesis that p150 is a master regulator of three-dimensional interactions of rDNA repeats. We will compare these data to genome-scale analyses of p150´s transcriptional targets and genomic localization. Together, these studies will provide candidate direct targets of p150 regulation, and lead us to test dependency relationships for these observations. Ribosomes, the cellular machines that synthesize proteins, contain large ribonucleic acid (RNA) molecules. We are pursuing our recent discovery that a protein previously thought to be dedicated to chromosome duplication is also an important regulator of ribosomal RNA production. Notably, ribosomal RNA synthesis also is regulated by cell growth and energy supply, as well as by tumor suppressors and oncoproteins, and is increasingly viewed as a therapeutic cancer target. Therefore, these studies will help us understand clinically relevant interactions that link DNA replication, gene expression, and growth control in human cells
Keywords: Binding (Molecular Function); cancer diagnosis; Cancer Prognosis; Cell Cycle; Cell Cycle Progression; Cell Cycle Regulation; cell growth; Cell Nucleolus; Cell Nucleus; Cell Proliferation; Cells; Chromatin; chromatin assembly factor I; chromatin immunoprecipitation; Chromatin Structure; Chromosomes; clinically relevant; Complex; Data; Data Set; Dependency (Psychology); Deposition; DNA; DNA biosynthesis; DNA Polymerase I; DNA Polymerase II; DNA Repair; Energy Supply; Epigenetic Process; Eukaryota; Event; fibrillarin; Gene Expression; Gene Expression Regulation; Genetic Transcription; Genome; Genome Stability; genome-wide; genome-wide analysis; Genomics; Growth; Heterochromatin; Histones; Human; in vivo; Label; Lead; Link; Maintenance; Malignant Neoplasms; Mass Spectrum Analysis; Mediating; Microscopy; Modification; Molecular; Molecular Conformation; novel; NPM1 gene; Nucleolar Proteins; nucleolin; nucleophosmin; Nucleoproteins; Nucleosomes; Oncogene Proteins; Physiologic pulse; prevent; Process; Production; Proliferation Marker; Promotor (Genetics); Protein Complex Subunit; Proteins; Recombinant DNA; Regulation; research study; restoration; Ribosomal RNA; Ribosomes; RNA; RNA chemical synthesis; RNA Interference; Role; Set protein; Site; small hairpin RNA; Structure; Tertiary Protein Structure; Testing; Therapeutic; Transcript; Transcriptional Activation; Transcriptional Regulation; Tumor Suppressor Proteins
Relevance: Ribosomes, the cellular machines that synthesize proteins, contain large ribonucleic acid (RNA) molecules. We are pursuing our recent discovery that a protein previously thought to be dedicated to chromosome duplication is also an important regulator of ribosomal RNA production. Notably, ribosomal RNA synthesis also is regulated by cell growth and energy supply, as well as by tumor suppressors and oncoproteins, and is increasingly viewed as a therapeutic cancer target. Therefore, these studies will help us understand clinically relevant interactions that link DNA replication, gene expression, and growth control in human cells
Project start date: 1997-06-01
Project end date: 2015-07-31
Budget start date: 1-AUG-2011
Budget end date: 31-JUL-2012
PFA/PA: PA-10-067
2R01GM055712-14A1 (2011): $398913
D Paul, Professor
University Of Pittsburgh At Pittsburghcity: Pittsburgh country: United States (us)
Grant 5R01AR051456-04 from National Institute Of Arthritis And Musculoskeletal And Skin Diseases
Abstract: Bone healing is complex process involving a number of different factors including soluble factors such as bone morphogenetic proteins (BMPs), signaling / transcription factors, nuclear transcription factors as well as extracellular matrix components. Although the delivery of BMPs as recombinant proteins can induce local bone formation and healing of bone defects, we and others have demonstrated previously that local gene transfer of BMP-2 at the site of critical size femoral and cranial defects in the rabbit and rat resulted in more rapid and efficient bone healing. More recently, we have shown that gene transfer of the LIM Mineralization Protein (LMP), a novel intracellular positive regulator of the osteoblast differentiation program, can induce efficient bone formation. In humans, three different LMP splice variants have been identified, termed LMP-1, LMP-2, and LMP-3. Gene transfer of human LMP-1 and LMP-3 induces expression of genes involved in bone formation including certain bone morphogenetic proteins (BMPs), promotes bone nodule formation in vitro and ectopic bone formation in vivo, facilitates healing of rat segmental and mandibular bone defects critical size defects and can facilitate posterior thoracic and lumbar spine fusion healing. We also have shown that LMP can induce myogenesis, under certain conditions, in culture. We have demonstrated that at least four different regions of LMP-1 and an additional domain in LMP-3 can contribute to osteogenesis. In partiular, a 20 amino acid region in LMP-3, termed Osteoinductive Domain-1 (OD-1), is able to induce mineralization and bone specific gene expression in cell culture and confer induction of ectopic bone formation in vivo. A synthetic OD-1 peptide fused to a protein transduction domain also was able to induce mineralization and bone specific genes. Thus the goals of this proposal are to identify the minimal as well as optimal LMP-1 and LMP-3 domains required for osteogenesis and myogenesis using cell culture assays, to examine the pathways through which the minimal, domains in LMP are able to induce osteogenesis (BMP signaling, RunX2/OSX transcription and/or MAP kinase) and to examine the ability of the LMP-1 and LMP-3-derived domains to induce efficient and appropriate bone formation in vivo following protein-transduction mediated delivery. The successful completion of the proposal experiments will lead to a better understanding of the pathways important for induction of osteogenesis by LMP and will result in clinically relevant approaches to stimulate new bone formation using LMP-derived peptides. Bone healing is complex process involving a number of different factors. Although the delivery of BMPs as recombinant proteins can induce local bone formation and healing of bone defects, we have demonstrated previously that local gene transfer of BMPs results in more rapid and efficient bone healing. We have shown that gene transfer of the LIM Mineralization Protein (LMP) can induce osteogenesis and bone formation as or more efficiently than BMP-2. The goals of this proposal are to identify the domains in LMP required for osteogenesis, to begin to examine the mechanisms through which LMP is able to induce osteogenesis and to examine the ability of the LMP-based domains to induce bone formation in vivo following gene and protein- transduction mediated delivery. The successful completion of the proposal experiments will lead to a better understanding of the pathways important for induction of osteogenesis by LMP and will result in clinically relevant approaches to stimulate new bone formation using LMP peptides
Keywords: adenoviral-mediated; Affect; Amino Acids; Autologous; base; Biological Assay; bone; bone healing; bone morphogenetic protein 2; Bone Morphogenetic Proteins; Cell Culture Techniques; Cephalic; Chest; Chimeric Proteins; clinically relevant; Complex; Defect; Dermal; Dose; Extracellular Matrix; Fibroblasts; Gene Expression; Gene Proteins; Gene Transfer; Genes; Genetic Transcription; Goals; Healed; healing; Health; Human; implantation; improved; In Vitro; in vivo; in vivo Model; inhibitor/antagonist; kinase inhibitor; Lead; Length; LIM Domain; Mandible; Maps; Mediating; Mesenchymal Stem Cells; Methods; mineralization; Mitogen-Activated Protein Kinases; Modeling; Mus; Muscle; myogenesis; Nodule; novel; Nuclear; Oryctolagus cuniculus; osteoblast differentiation; Osteogenesis; osteogenic; overexpression; Pathway interactions; Peptides; Physiologic calcification; Play; Process; programs; Promotor (Genetics); Protein Fragment; Protein Splicing; Proteins; Rattus; Recombinant Proteins; research study; RNA Interference; Role; scaffold; Signal Transduction; Site; synthetic peptide; Tertiary Protein Structure; transcription factor; Variant; Vertebral column; yeast two hybrid system; Yeasts
Relevance: Bone healing is complex process involving a number of different factors. Although the delivery of BMPs as recombinant proteins can induce local bone formation and healing of bone defects, we have demonstrated previously that local gene transfer of BMPs results in more rapid and efficient bone healing. We have shown that gene transfer of the LIM Mineralization Protein (LMP) can induce osteogenesis and bone formation as or more efficiently than BMP-2. The goals of this proposal are to identify the domains in LMP required for osteogenesis, to begin to examine the mechanisms through which LMP is able to induce osteogenesis and to examine the ability of the LMP-based domains to induce bone formation in vivo following gene and protein- transduction mediated delivery. The successful completion of the proposal experiments will lead to a better understanding of the pathways important for induction of osteogenesis by LMP and will result in clinically relevant approaches to stimulate new bone formation using LMP peptides
Project start date: 2008-09-10
Project end date: 2013-08-31
Budget start date: 1-SEP-2011
Budget end date: 31-AUG-2012
PFA/PA: PA-07-070
5R01AR051456-04 (2011): $311673
DISABILITY AND HEALTH OUTCOMES IN COPD
D Paul
University Of California San Franciscocity: San Francisco country: United States (us)
Grant 5R01HL077618-05 from National Heart, Lung, And Blood Institute
Abstract: Background. COPD is a common chronic health condition. Because current medical treatments have minimal impact on disease progression, a strategy to prevent COPD-related disability would have important clinical and public health benefits. Study Aims. To elucidate the disablement process, we will test a specific conceptual model of how disability develops in COPD. The aims are (1) To evaluate the impact of respiratory impairment, especially pulmonary function impairment, on the risk of functional limitation in COPD. Using a control group, to elucidate the prevalence of respiratory impairment, functional limitation, and disability that is directly attributable to COPD. (2) In adults with COPD, to delineate the longitudinal effect of functional limitation on the risk of incident disability. We hypothesize that development of functional limitation, and not pulmonary function impairment, is the major determinant of disability. (3) To determine the prospective impact of disability on the risk of future adverse health outcomes. Methods. We will assemble a prospective cohort of 1200 randomly sampled adults with COPD who are members of a large regional health maintenance organization. A matched control group of 300 subjects will be recruited. Subjects will undergo a detailed physical assessment that measures respiratory impairment (e.g., pulmonary function) and functional limitation (e.g., lower extremity function, muscle strength, exercise performance, and cognitive function). Structured telephone interviews will ascertain disability outcomes at baseline and 18-month prospective follow-up. We will also study the modulating effects of specific risk factors and protective factors on the progression from functional limitation to COPD-related disability, including psychological factors, environmental exposures, and health care process factors. Significance. Elucidation of the disablement process is the first step toward designing clinical and public health interventions that target reduction of disability in COPD. By determining the pathway from functional limitation to disability, our goal is to provide a scientific basis for screening and prevention of disability among persons with COPD. Based on this work, intervention strategies can be designed that target reduction of functional limitation and disability to improve downstream health outcomes
Keywords: 21+ years old; Active Follow-up; Adrenal Cortex Hormones; Adult; adult human (21+); adult youth; Affect; Age; aged; Anxiety; Aspiration, Respiratory; base; Breathing; Cardiovascular Diseases; cardiovascular disorder; Care, Health; cease smoking; Cessation of smoking; Chronic; Chronic Obstructive Airway Disease; Chronic Obstructive Lung Disease; cigarette smoking; Clinical; COAD; Cognitive decline; Cognitive Disturbance; cognitive dysfunction; cognitive function; Cognitive function abnormal; Cognitive Impairment; cognitive loss; cognitively impaired; cohort; Condition; Control Groups; COPD; Corticoids; Corticosteroids; Daily; Depression; design; designing; Deterioration; Development; diabetes; Diabetes Mellitus; disability; Disease Progression; disease severity; Disturbance in cognition; Early Intervention; Early Intervention (Education); Economic Income; Economical Income; Education; Educational aspects; Environmental air flow; Environmental Exposure; Ethnic Origin; Ethnicity; Ethnicity aspects; Exercise; Exercise, Physical; follow-up; functional disability; Functional impairment; Future; Goals; Health; Health Benefit; Health Maintenance Organizations; health related quality of life; Healthcare; HMO; HOSP; Hospitalization; Human, Adult; Impaired cognition; Impairment; improved; Income; Indoor Air Pollution; Inhalation; Inhaling; inspiration; Inspiration, Respiratory; Intervention; Intervention Strategies; interventional strategy; Job Environment; Job Location; Job Place; Job Setting; Job Site; Link; Lower Extremity; Lower Limb; Lung diseases; lung disorder; Measures; Medical; member; Membrum inferius; Mental Depression; Methods; Modeling; Multivariate Analyses; Multivariate Analysis; muscle strength; Muscle Weakness; Muscular Weakness; Nicotine; O element; O2 element; Outcome; Oxygen; Pathology; pathway; Pathway interactions; Patients; Performance; Persons; Physical assessment; Population; Prepaid Group Health Organizations; Prevalence; prevent; preventing; Prevention; Prevention strategy; Preventive strategy; Process; prospective; Psychological Factors; Public Health; public health medicine (field); Pulmonary Disease, Chronic Obstructive; Pulmonary Diseases; Pulmonary Disorder; pulmonary function; Pyridine, 3-(1-methyl-2-pyrrolidinyl)-, (S)-; Race; Racial Group; recruit; Recruitment Activity; respiratory; Respiratory Disease; Respiratory Disorder; Respiratory System Disease; Respiratory System Disorder; Risk; Risk Factors; Sampling; screening; Screening procedure; screenings; Self Efficacy; Severity of illness; sex; smoke cigarette; smoking cessation; Social support; social support network; Socio-economic status; Socioeconomic Status; Staging System; Status, Socioeconomic; Stocks, Racial; Structure; Telephone Interviews; Testing; Therapeutic Corticosteroid; Tobacco; Vaccination; Ventilation; Work; work environment; Work Location; Work Place; work setting; Work-Site; Workplace; Worksite; young adult
Project start date: 2004-07-01
Project end date: 2011-06-30
Budget start date: 1-JUL-2008
Budget end date: 30-JUN-2011
5R01HL077618-05 (2008): $504153
D Paul
Yale Universitycity: New Haven country: United States (us)
Abstract: ADMINISTRATIVE (ADMIN) CORE The Admin Core is the Center´s planning and decision-making body and provides overall scientific direction and leadership. It will serve as CIRA´s central administrator, fiscal executor and coordinating body; ensure that the Center is a primary source of information on HIV/AIDS related topics; monitor Center activities and develop, and implement and evaluate annual strategic plans. A Director, two Deputy Directors, an Executive Director, and two Assistant Directors, will comprise the Center´s Leadership Group (LG), which will meet monthly to guide its administrative and scientific direction and activities. The LG will meet semi-annually with all Core Directors to discuss Core activities and plans and financial resources for implementing them, monitor Core progress in following its strategic plan, and facilitate coordination and collaboration among Core Directors. The Core will convene four advisory bodies to provide guidance in its activities. The Executive Committee will advise on all aspects of the Center´s operations, including Center policies, priorities, and future scientific directions. The External Advisory Board will bring together individuals of national prominence to review the Center´s progress and plans on an annual basis and provide advice on its future development. The Deans´ Committee will advise on how CIRA can best involve scientists from different Yale graduate and professional schools and establish linkages throughout the University. The Community Advisory Board, comprised of representatives from community agencies and individuals infected and affected by HIV/AIDS, will help to ensure both the relevance of the Center´s research to the needs of the community and the input of the community in CIRA´s research. The Admin Core will oversee the affiliation process and help facilitate CIRA science by providing a range of services, including grant preparation and, for CIRA scientists with primary appointments in the School of Public Health, grant management; assistance with personnel planning; development and maintenance of centralized databases for information sharing, program monitoring, and strategic planning; publication of the weekly email bulletin; and development and maintenance of CIRA´s internet and intranet websites. The Admin Core will also organize CIRA´s AIDS Science Day
Keywords: Acquired Immunodeficiency Syndrome; Administrator; Affect; AIDS/HIV problem; Appointment; base; Communities; Coordination and Collaboration; Databases; Decision Making; Development; Electronic Mail; Ensure; Epidemic; Foundations; Future; Grant; Human Resources; Individual; Interdisciplinary Study; Internet; Intranet; Leadership; Link; Maintenance; meetings; Mission; Monitor; operation; Policies; Preparation; Process; programs; Public Health Schools; Publications; Research; Research Infrastructure; Resources; Schools; Science; Scientist; Services; sound; Source; Strategic Planning; Universities; web site
Budget start date: 1-JUL-2011
Budget end date: 30-JUN-2012
5P30MH062294-09_9001 (2011): $684922
HETEROZYGOUS MH KNOCK-IN MICE MODEL HUMAN MH SUSCEPTIBILITY
D Paul, Professor
Brigham And Women´s Hospitalcity: Boston country: United States (us)
Budget start date: 1-APR-2010
Budget end date: 31-MAR-2011
5P01AR052354-05_0001 (2010): $228355