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Record Count: 105
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DESCRIPTION (provided by applicant): Several different in vitro and ex vivo-derived rodent cell lines were developed that recapitulate the specialized asymmetric cell kinetics of adult somatic tissue stem cells in vivo. Previously, these model cells have been used to investigate and define genetic and molecular mechanisms that control adult stem cell kinetics. In the proposed research, the model cells will be used to investigate whether environmental agents, that are known to induce human cancers but with no detectable mutagenic activity, do so by disrupting adult stem cell kinetics. Specifically, the hypothesis that some non-mutagenic carcinogens function by inducing increased rates of stem cell proliferation will be examined. Disruption of asymmetric stem cell kinetics is an essential step in human carcinogenesis. "Kinetotoxic" effects by environmental contaminants, that lead to increased adult stem cell proliferation, may also be important in the etiology of other chronic processes associated with altered cell proliferation (e.g., premature aging). The model cells have already been used to identify 3 organic compounds that switch exposed cells from asymmetric cell kinetics to exponential cell kinetics. The main focus of the proposed research will be to use these cell lines to evaluate well established non-mutagenic environmental carcinogens for kinetotoxicity. Both acute high-dose and chronic low-dose exposures will be investigated. In addition to evaluating non-mutagenic compounds, several well-known mutagenic carcinogens will be examined, as they may exhibit concurrent kinetotoxicity. Chemicals found to induce changes in cell kinetics will be evaluated for molecular effects on cellular genes previously identified as regulators of asymmetric cell kinetics. In preliminary studies, the IARC Group 1 human carcinogen benzene exhibits evidence for kinetotoxicity. Benzene has a well-documented history as a potent non-mutagenic human carcinogen. It will serve as the prototype kinetotoxic agent for developing a systematic approach to revealing kinetotoxic mechanisms. In vitro human cell lines with asymmetric cell kinetics will be derived to provide kinetotoxicity assays that may have greater relevance to human adult stem cell physiology. The proposed research has the goal of providing new insights to the carcinogenic mechanisms of well studied environmental contaminants whose mode of action remains a mystery. Such insights will advance efforts to develop better strategies to limit morbidity associated with exposure to environmental carcinogens.
DESCRIPTION (provided by applicant): The broad long-term objective of this proposal is to elucidate the molecular basis of the Benzofalpyrene (B[a]P)-induced S-phase checkpoint. B[a]P is an abundant and ubiquitous environmental carcinogen that is metabolized intracellularly to generate Benzo[a]pyrene Di-hydrodiol-Epoxide (BPDE). BPDE-induced DMA damage triggers S-phase checkpoint signaling pathways that coordinate replication and repair of the genome. However, the molecular mechanism(s) by which checkpoint signaling regulates DNA replication are incompletely understood. Inaccurate repair and replication of damaged DNA can result in genomic instability, a hallmark of cancer cells. The BPDE-induced S-phase checkpoint is likely to help maintain genomic stability and prevent cancer after B[a]P exposure. This application will investigate the mechanisms by which S-phase checkpoints are activated by DNA damage, and the Trans-Lesion Synthesis (TLS)-mediated mechanisms that enable recovery from the checkpoint. Our studies indicate that Cdc45 (a DNA replication factor) and DNA polymerase Pol kappa (Pol?) are involved in S-phase checkpoint activation and recovery respectively. We have also shown that Pol? is regulated by the E3 ubiquitin ligase Rad18. The Specific Aims of this proposal are: (1) To test the hypothesis that Cdc45 is a target of the BPDE-induced S- phase checkpoint. (2) To elucidate the role of Rad18 in recovery from the BPDE-induced S-phase checkpoint. (3) To elucidate the role of the Pol?-interacting DNA polymerase Rev1 in checkpoint recovery. Aims 1 and 2 will identify and mutate sites of Cdc45 ubiquitination (Aim 1) and Rad18 phosphorylation (Aim 2). Then we will study the regulation of ubiquitination and phosphorylation-resistant mutants by checkpoint signaling. These experiments will test the significance of Cdc45 ubiquitination and Rad18 phosphorylation in relation to the S-phase checkpoint. In Aim 3 we will determine the effect of ablating Rev1 (which interacts physically and genetically with Pol?) on Pol? regulation. These studies will determine how interactions between Pol? and Rev1 regulate the S-phase checkpoint. Together, these experiments will provide a novel paradigm for the mechanisms by which the DNA replication machinery is coordinated with DNA repair proteins in response to B[a]P (and .possibly other genotoxins) to maintain genomic stability. Results of our studies might help identify individuals that are at high-risk for environmental B[a]P-induced disease. Moreover, our studies could help identify novel drug targets for cancer therapy: Similar to B[a]P, many chemotherapies are genotoxic and activate checkpoint pathways. We have shown that Pol? or Rad18- deficiency sensitizes cells to B[a]P-induced death. Potentially, small molecules that target Rad18, Pol?, or other TLS enzymes could sensitize cancer cells to killing by chemotherapeutic agents.
DESCRIPTION (provided by applicant):
It is widely appreciated that environmental stresses, such as exposure to toxins or pathogens, can have profound impact on individual health and well-being. How these "stressors" exert their influence is less well understood. While specific targets of environmental stresses have been identified for some toxins and pathogens, we propose that the impact of environmental stresses is mediated through the pathway(s) and/or networks in which the target molecule participates. Furthermore, even where a causal association between an environmental stress and a biological target has been identified, the biological insight that must precede a strategy for therapeutic intervention has generally been slow in coming. We suggest that the phenotypic effects of environmental stresses are mediated by alterations in a dynamic network of gene products and metabolites, and such networks, normal and perturbed, exhibit emergent properties that cannot be understood one gene at a time. Our central hypothesis is that one must understand changes in complex cellular networks to fully understand the link between genotype, environment, and phenotype. The Center for Cancer Systems Biology (CCSB) at the Dana-Farber Cancer Institute, together with a growing number of laboratories, is mapping and modeling cellular pathways and networks both "locally" at the scale of "molecular circuits" made of a few interacting molecules, and "globally" at the scale of the whole proteome; fundamental biological insights have already emerged from this body of work. We propose to use a model system as a surrogate for generalized environmental stresses to begin understanding how cellular pathways and networks are altered or modified as a consequence of environmental insult. We have chosen a defined, model system with a variety of disease outcomes: viral infection. Here we explore the concept that environmental stresses, as exemplified by viruses, influence local and global properties of networks to induce "disease states". Our plans to achieve these goals are summarized in the following specific aims: 1) Profile all binary viral-host protein-protein interactions for a group of viruses with related biological properties 2) Systematically test each viral protein for its ability to disrupt or alter host-host protein interactions, 3) Integrate the resulting interaction and perturbation data with diverse genomic data sources to derive dynamic cellular network models.
DESCRIPTION (provided by applicant): The long-term goal of our laboratory is to understand, in molecular detail, how protein kinase signaling pathways together with phosphoserine/threonine-binding domains regulate multiple aspects of cell proliferation, including cell cycle progression and the cellular response to DNA damage. In the present proposal, we explore the function of MAPKAP Kinase-2, a stress-responsive protein kinase activated by p38 MAPK, as a critical regulator of S-phase and mitotic progression in response to environmental and endogenous types of DNA damage. We use a combination of extensive biochemistry and molecular cell biology to explore the signal transduction mechanisms involved in MAPKAP Kinase-2 activation after DNA damage induced by chemicals and UV-C irradiation, and examine how MAPKAP Kinase-2 functions together with other checkpoint kinases such as Chk1, to control cell cycle progression after genotoxic stress in cells in culture. We go on to develop a conditional MAPKAP Kinase-2 knock-out mouse to explore whether MAPKAP Kinase-2 functions as a tumor suppressor in genetically defined models of sarcoma and lung cancer, and in environmental carcinogen-induced models of colorectal and skin cancer. Finally, we explore whether down-regulation of MAPKAP Kinase-2 facilitates cell death after intentional chemically-induced DNA damage such as chemotherapy. These studies should clarify how signals from the p38 MAPK-MAPKAP Kinase-2 pathway, a global stress-responsive network that is activated by a wide variety of toxic insults, integrate with those from dedicated DNA damage response pathways, to regulate the cellular response to genotoxic stress. The results of the proposed experiments should reveal whether MAPKAP Kinase-2 functions as a tumor suppressor gene that modulates the risk of cancer after exposure to environmental agents, and whether specific targeting of MAPKAP Kinase-2 would be of therapeutic value for sensitizing tumors to the cytotoxic effects of conventional chemotherapy.
Crisp Terms/Key Words: phosphoserine, laboratory mouse, genetically modified animal, biochemistry, biological signal transduction, cell death, cell cycle, enzyme structure, disease /disorder model, molecular biology, sarcoma, lung neoplasm, protein kinase, protein structure function, DNA damage, mitogen activated protein kinase, cell proliferation, tumor suppressor protein, binding site, serine threonine protein kinase
DESCRIPTION (provided by applicant): Our long-term objective is to determine the mechanisms by which developmental exposure to dioxins and other ubiquitous arylhydrocarbon receptor (AhR) ligands alters reproductive functions in adulthood. We recently showed that a single developmental exposure to the potent AhR ligand, 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD), interferes with estrogen-dependent sexual differentiation of the male anteroventral periventricular nucleus (AVPV). The AVPV controls the female pattern of luteinizing hormone (LH) release; therefore, TCDD-exposed males have a female-like AVPV structure and they show the female, cyclic pattern of LH release in adulthood. Considering that estradiol (E2) derived from fetal production of testosterone is responsible for the sexual differentiation process, we hypothesize that TCDD interferes with E2 regulation of a set of genes during development. To test this hypothesis, we will use Affymetrix microarrays and cluster analysis to identify sex-specific genes that are regulated by E2 and TCDD. We will use a "redundancy model", testing different animal treatments predicted by our hypothesis to target the same set of genes. We will validate our microarray findings using RT-QPCR, in situ hybridization histochemistry, developmental ontogeny studies, dose response studies and promoter analysis. Our findings will be important for identifying genes underlying effects of dioxins and other ubiquitous AhR ligands on reproductive toxicity, as well as on other neural functions disrupted by perinatal TCDD exposure in a sex-specific manner. Such findings will be important for mechanism-based risk assessment, as well as for development of pharmaceutical interventions to prevent neurotoxicity.
DESCRIPTION (provided by applicant):
Associations between breast cancer and exposure to environmental carcinogens are often weak or inconsistent in the epidemiologic literature, even when strong experimental data exists. The strongest support for environmental carcinogenesis comes from mouse models showing that exposures such as ionizing radiation (IR) and polycyclic aromatic hydrocarbons (PAHs) increase mammary tumor formation. These same experimental models have shown that parity has a p53-mediated, protective effect against carcinogenesis. This application proposes a comparative toxicology approach for dissecting the biological pathways that lead to environmental carcinogenesis of the breast, with emphasis on interactions between environmental carcinogens, the p53 pathway, and parity. Three classes of environmental exposure, each with a strong epidemiologic and toxicological literature, have been selected for study: IR, PAHs, and organochlorines (OCs). The first two classes (IR and PAHs) both cause DNA damage and induce p53-dependent DNA damage responses, but the DNA damage mechanisms are different. In contrast, OCs do not act by directly damaging DNA, and thus, provide a negative control for p53-dependent signaling. OCs also provide a comparison for the PAHs as both PAHs and OCs can activate the aryl hydrocarbon receptor (AhR) and perturb endocrine action in the breast. In Aim 1, we will assess the p53 dependent effects of each toxicant using isogenic human breast cell lines (with and without expression of p53RNAi to knockdown p53 expression) and nulliparous Trp53+/+ and Trp53-/- mice. In Aim 2, we will examine gene expression profiles associated with parity in humans (using reduction mammoplasty samples) and mice, with specific attention to identifying p53-regulated gene expression. In Aim 3, the interaction between parity and environmental carcinogenesis will be assessed by comparing transcriptional responses to toxicants in parous and age-matched nulliparous mice. Differential responses to toxicants will be verified in explant cultures of human breast tissue from nulliparous and parous women (excess tissue from reduction mammoplasty). Thus, each aim will integrate human and mouse responses to define responses to toxicants and parity. The data from all three aims will be integrated to allow a complete, multifactorial evaluation of how environmental exposures to these toxicants, p53 signaling, and parity status interact to affect breast signaling, and ultimately carcinogenesis. These studies will define the mechanisms of environmentally-induced breast cancer, anchored on genotype-phenotype correlations that define susceptibility, and will identify new candidate biomarkers of susceptibility that can be used in future animal and human studies.
DESCRIPTION (provided by applicant): Within the broad goal of identifying biomarkers of human disease, the objective of the proposed studies is to define the chemical basis for the formation of endogenous DNA, RNA and protein adducts, with a focus on the roles of deoxyribose and lipid oxidation in generating these lesions. We propose to build on our observation of DNA and protein adducts derived from deoxyribose oxidation by moving the studies into living cells and tissues, by extending the studies to RNA as a potential target for reactive electrophiles, and by applying new technologies to study the DNA, RNA and protein adducts formed by analogous electrophiles arising from lipid peroxidation. Altogether, the four specific aims of the proposed studies should provide new insights into the chemical basis for endogenous nucleic acid and protein adducts that may be useful as biomarkers of exposure and disease. Aim #1) Protein and DNA adducts derived from 4'-oxidation of deoxyribose. We will continue to study the role of base propenals and lipid peroxidation in the formation of the M1dG adduct in DNA and RNA in human cells, and expand 4'-oxidation adduct studies to include the 2-deoxypentose-4-ulose abasic site. Aim 2) Protein and DNA adducts derived from 5'-oxidation of deoxyribose. We will move our studies of the novel DNA adducts derived from 2-phosphoryl-1,4-dioxobutane, a product of deoxyribose 5'-oxidation, into cells and tissues of animal models of human disease and expand the studies to include lysine adducts in histone proteins. Aim 3) Histone adducts derived from 3'-formylphosphate residues. We will continue our studies of lysine N6- formylation in histones by proteomic mapping of the adduct locations, by LC/MS/MS quantification of adducts in cells and tissues, and by assessing the reactivity of histone deactylases with formyllysines in peptides. Aim 4) Lipid peroxidation as a source of DNA, RNA and protein adducts. Given the parallels between deoxyribose and lipid oxidation, we propose to extend our deoxyribose studies by surveying the extent of protein, DNA and RNA adduct formation by lipid peroxidation products by incorporating 14C-labeled PUFA into cell membranes followed by quantification of 14C in protein, DNA and RNA fractions and identification of the DNA and RNA adducts. Subsequent efforts will be directed at defining structures of major adducts.
DESCRIPTION (provided by applicant):
The increased incidence of uterine leiomyoma, testicular and breast cancer observed in European and US populations during the last 50 years prompted scientists to hypothesize that prenatal exposure to environmental estrogens may be the underlying cause of these neoplasms. Epidemiological studies suggest that fluctuating estrogen levels in the fetal environment have long-term consequences regarding the risk of developing breast cancer during adult life. This is supported by laboratory rodent studies showing that perinatal exposure to pharmacological doses of diethylstilbestrol (DES) increases the incidence and decreases the latency period of mammary cancer. Among the xenoestrogens, bisphenol-A was chosen for these studies because of its widespread use and because it is found in 95% of the urine samples analyzed in a recent CDC study. Perinatal exposure to environmentally relevant doses of the xenoestrogen bisphenol A (BPA) alters the development of the rodent mammary gland and results in outcomes such as increased sensitivity to estrogens at puberty, increased ductal density in adulthood, and an increase in the number of structures where cancer arises. Using a rat mammary carcinogenesis model a pilot experiment was performed that revealed that perinatal exposure to low BPA doses induces intraductal hyperplasias. When challenged with a subcarcinogenic dose of nitrosomethylurea, only the BPA exposed animals developed neoplasias. Thus it is hypothesized that BPA administered perinatally will permanently alter tissue-specific patterns of gene expression by altering the methylation pattern of specific genes. These alterations will, in turn, affect development beyond the period of exposure to BPA, leading to an increase in susceptibility to breast cancer. Specific Aim 1 will compare the global pattern of DNA methylation in the mammary glands of vehicle and BPA-exposed animals and to construct a chromosomal methylation idiogram. The three components of Specific Aim 2 are to: 1) Determine the mRNA expression profiles of the BPA-treated mammary glands by DNA microarray analysis and to map the loci of differentially expressed genes on the rat chromosomal ideogram; 2) Identify candidate genes that are differentially expressed according to the mRNA microarray analysis that map onto the BPA-methylated or demethylated chromosomal regions; and 3) Narrow the set of candidate genes by bioinformatics techniques (molecular pathway and ontology analysis). The most promising candidate(s) will be probed in detail using bisulfite mapping of the methylated CpG dinucleotides. If a causal link is found between perinatal BPA exposure and alterations in gene methylation, this work will provide testable hypotheses connecting BPA-induced gene marking and propensity to develop mammary gland neoplasia. If the hypothesis is consistent with the result, this will constitute a major shift in emphasis from adult exposures to in utero exposures. This will also have a great impact on the way we study risk factors and conduct epidemiological studies; it may even influence public policy about breast cancer prevention.
Crisp Terms/Key Words: epigenetics, bioinformatics, microarray technology, gene environment interaction, mammary gland, phenol, messenger RNA, DNA methylation, breast neoplasm, environment related neoplasm /cancer, antiantibody, immunoprecipitation, genetic regulation, gene expression, genetic mapping, estrogen analog, laboratory rat
DESCRIPTION (provided by applicant):
Cardiovascular disease (CVD) is the leading cause of death among the elderly in the United States. Since the percentage of Americans over the age of 75 years is expected to increase from 6% in 2000 to 11 % in 2050, the number of persons dying from CVD will likely rise dramatically in the coming years. In previous work, this team of investigators demonstrated that acute and chronic particulate air pollution exposures are associated with specific cardiovascular outcomes, such as decreased heart rate variability, arterial stiffness, and ST-segment depression, resulting in increased death rates for CVD.
DNA-methylation changes have been found to be involved in inflammation, endothelial function, and atherosclerosis. Initial observations in experimental models and humans have shown that air particles and metals, which are constituents of particulate pollution, alter DNA methylation status. Whether the increased risk of CVD observed in subjects exposed to air particles and toxic metals (such as arsenic, cadmium and lead) is mediated by alterations of DNA-methylation patterns is unresolved. The goal of this proposal is to determine whether exposure to air particles and toxic metals (arsenic, cadmium and lead) increases CVD risk by altering global and/or gene specific methylation in elderly subjects over time. Specifically, this proposal will evaluate: 1) the association of the exposure with global DNA methylation in Alu and LINE-1 sequences, and with hypermethylation or hypomethylation in specific genes (ICAM-1, IFN-gamma, p16) involved endothelial function and atherosclerosis; and 2) the association between the above global and gene specific DNA-methylation changes and CVD-related outcomes (Heart-rate variability, Pulse Doppler measurements of atherosclerosis, ST segment elevation/depression, biological markers of inflammation/endothelial function). As a secondary aim, this initiative will explore whether: 1) global or gene-specific methylation status modifies the association of air particles and toxic metals (Pb, As, Cd) with CVD-related endpoints; and 2) the association between exposure and methylation status is modified by plasma levels or intakes of dietary factors (folate, Vitamin B6 & B12), plasma homocysteine levels, and SNPs in DNA methylation genes (MTHFR, cSHMT). The study design, based on an existing cohort of 700 elderly subjects in Eastern Massachusetts, will allow for baseline and longitudinal analysis of air particles, metal biomarkers, DNA methylation patterns and CVD phenotypes, therefore all hypotheses will be tested cross-sectionally and longitudinally. Quantitative DNA methylation analysis, based on PCR amplification and pyrosequencing of bisulphite-treated DNA, will be performed for each study subject on three blood DNA samples collected over a long time span.
DESCRIPTION (provided by applicant): The Reference Epigenome Mapping Centers (REMC) will aim to transform our understanding of human epigenetics through production and integrative analysis of comprehensive reference epigenomes for ES cells, differentiated cells and tissues. In pursuit of this goal, we have assembled a unique scientific team and infrastructure with broad expertise and capabilities in stem cell biology, epigenomics, technology, production research and computation. We recently demonstrated two complementary methods that leverage ultra high-throughput sequencing for epigenomic analysis. In the first method, genome-wide chromatin maps are acquired by deep sequencing chromatin IP DNA (ChlP-Seq). In the second, nucleotide-resolution DNA methylation maps are generated by high-throughput bisulfite-sequencing (HTBS). These methods represent major improvements over prior tools as they yield precise digital information, have high genome coverage, require fewer cells and are cost-effective. Multiple epigenomic maps have already been produced for stem cells and primary tissues, and pipelines have been assembled for efficient data collection, processing and analysis. For the REMC project, we propose to apply ChlP-Seq and HTBS pipelines to generate comprehensive high-resolution maps of chromatin state and DNA methylation for 100 diverse cell types. Cell types were selected for their biological and medical importance, and for their potential to maximize the comprehensiveness of acquired epigenomic data. They include human ES cells, ES-derived cells, mesenchymal stem cells, reprogrammed stem cells and primary tissues. ChlP-Seq will be used to map highly informative chromatin modifications and related chromatin proteins in each cell type. HTBS will be used to generate nucleotide-resolution DNA methylation maps. Reference epigenomes will reveal the locations and activation states of diverse functional genomic elements, inform on the developmental state and potential of studied cell populations, and provide a framework for understanding complex epigenetic regulatory mechanisms. All data will be made available to the scientific community upon verification. PUBLIC HEALTH RELEVANCE: Comprehensive characterization of epigenetic marks ('the epigenome') is a critical step towards a global understanding of the human genome in health and disease. The proposed mapping studies will provide unprecedented views of the human epigenetic landscape and its variation across cell states, offer fundamental insight into the functions and interrelationships of epigenetic marks, and provide a framework for future studies of normal and diseased epigenomes.
DESCRIPTION (provided by applicant): DMA repair and damage tolerance processes are absolutely critical to preserving human health following exposure to many different agents. The long term goal of this research is to develop a detailed integrated understanding of the molecular mechanisms responsible for environmental mutagenesis in eukaryotes. In molecular processes that are both complex and elaborately controlled, mutations are introduced when specialized translesion synthesis (TLS) DNA polymerases copy over DNA damage caused by environmental agents. The proposed research places a special emphasis on Rev1, which by virtue of acting both as a scaffold for other TLS DNA polymerases and as a polymerase itself, lies at the root of eukaryotic mutagenesis. We will follow up on our unanticipated finding that S. cerevisiae Rev1 is expressed 50 fold higher during G2/M than in G1 and most of S phase by investigating the basis of its cell cycle control," which is predominantly posttranscriptional; promising candidate regulatory genes such as UMP1 and CDC7 will be tested, genetic screens for new regulatory genes will be carried out, and experiments will be conducted to determine the importance Of this control: Using both biochemical and genetic approaches, we will continue to investigate the structural and functional basis of Rev1 interactions, focusing particularly on the interactions of the Rev1 C-terminal domain, its BRCT domain, and its ubiquitin-binding motifs. We will investigate the functional importance of the Rev1 polymerase activity by following up on our recent observations suggesting that Rev1 may have a class of cognate lesions it replicates particularly well. We will investigate the function and regulation of S. pombe DinB and its relationship to Rev1. We will develop a high-throughput assay, based on disruption of the Rev1-Rev7 interaction, that will allow screening for inhibitors of eukaryotic environmental mutagenesis. The proposed research will make a highly significant contribution to basic science by elucidating the still poorly understood eukaryotic translesion synthesis mechanisms responsible for most mutations. These mutations contribute to aging, cancer, and various human diseases. Identification of additional genes that play roles in these mutagenic processes will help make it possible to address the question of why only some people develop disease when exposed to an environmental toxin. Small molecule inhibitors of TLS could lead to novel "anti-mutagenesis" drugs with multiple applications to human health.
DESCRIPTION (provided by applicant): The broad long-term objective of this proposal is to elucidate the role of the Fanconi Anemia (FA) pathway in protecting cells from the carcinogenic effects of Benzo[a]pyrene (B[a]P)-induced DNA damage. B[a]P is an abundant and ubiquitous environmental carcinogen that is metabolized intracellularly to generate Benzo[a]pyrene Di-hydrodiol-Epoxide (BPDE). BPDE-induced DNA damage triggers Rad18 (an E3 ligase)- dependent mono-ubiquitination of PCNA and recruitment of the specialized DNA polymerase Pol kappa to replication forks. In contrast with replicative DNA polymerases which stall at sites of DNA damage, Pol kappa can perform DNA synthesis using BPDE-adducted DNA as a template. Pol kappa-mediated TLS enables cells to tolerate the mutagenic and lethal effects of BPDE adducts, thereby contributing to genomic stability and tumor suppression. However, the molecular mechanism(s) of Pol kappa regulation are incompletely understood. Our preliminary studies suggest that the Fanconi Anemia (FA) proteins are intimately involved in Rad18 and Pol kappa- mediated TLS of B[a]P lesions. The Specific Aims of this proposal are: (1) To determine the molecular basis for BPDE-induced interaction between FANCD2 (the putative effector of the FA pathway) and Pol kappa. (2) To determine the mechanism by which Rad18 activates the FA pathway. (3) To test the interdependence of the FA and TLS pathways. Aim 1 will mutate ubiquitin binding motifs in Pol kappa and determine the effect of these mutations on FANCD2 association. Additionally, we will test the role of the Pol kappa- and FA pathway-interacting protein REV1 in mediating Pol kappa-FANCD2 interactions. Aim 2 will test whether Rad18 modifies FANCD2 directly or via regulation of upstream FA core complex components. Aim 3 will determine whether FANC- deficiency compromises TLS of BPDE lesions and conversely, whether defective TLS affects activation of the FA pathway by BPDE. Results of our studies will provide a novel paradigm for mechanisms by which the FA pathway is coordinated with TLS enzymes in response to environmental B[a]P (and possibly other genotoxins) to maintain genomic stability. Our studies might help identify individuals that are at high-risk for environmental B[a]P-induced disease. Moreover, our work could lead to novel drug targets for cancer therapy: Similar to B[a]P, many chemotherapies are genotoxic and activate checkpoint pathways. We have shown that Pol kappa- or Rad18-deficiency sensitizes cells to B[a]P-induced death. Potentially, small molecules that target FA components, Rad18, Pol kappa, or other TLS enzymes could sensitize cancer cells to killing by chemotherapeutic agents. PUBLIC HEALTH RELEVANCE: Benzo[a]pyrene (B[a]P) is an abundant environmental pollutant that damages DNA thereby causing permanent changes to the genetic code (termed 'mutations') that can result in cancer. Our results suggest that a group of cellular proteins termed the 'Fanconi Anemia' proteins (FANCs) play a role in preventing environmental B[a]P- induced mutations. The proposed experiments aim to understand the mechanisms by which FANCs help cells tolerate the effects of environmentally-induced DNA damage and protect against cancer.
DESCRIPTION (provided by applicant): The potent environmental carcinogen benzo[a]pyrene (B[a]P) is metabolically activated in cells to (+)-anti-B[a]PDE, which forms 1 primary adduct: [+ta]-B[a]P-N2-dG. [+ta]-B[a]P-N2-dG induces different mutational patterns depending on sequence context (e.g., >95% G->T vs. 95% G->A in 5'-TGC vs. 5'-AGA sequences). Evidence suggests that these different mutations arise from different adduct conformations (as influenced by sequence context), when bypassed by different DNA polymerases. For [+ta]-B[a]P-N2-dG, we showed that E. coli DNA Pol V is involved in dATP bypass (G->T mutations), while dCTP insertion (no mutation) involves Pols IV and V. With the mirror image adduct [-ta]-B[a]P-N2-dG, Pol V does dATP insertion, while Pol IV alone is required for dCTP bypass. Literature findings suggest that in general DNA Pol V has 2 modes of adduct bypass: (1) correct dNTP insertion, and (2) default dATP insertion. Understanding the mechanism of correct vs. mutagenic insertion is hampered by no X-ray structure for UmuC, which is the polymerase subunit of DNA Pol V. Using homology modeling, we constructed a UmuC model, which revealed active site amino acids potentially involved in dictating dNTP insertion. Active site amino acids were changed. In cells we showed that mutant-UmuCs could increase (up to ~10-fold), or decrease (~5-fold) dATP insertion compared to wt-UmuC. The goal of this project is to understand what amino acid residues define correct (dCTP) vs. incorrect (dATP) insertion for Pols IV and V and how these pathways are controlled by the cell. Studies in cells and in vitro with mutant and wild type Pols IV and V are proposed. Literature findings show that Pol IV is equivalent to human Pol k, while Pol V is equivalent to human Pol h. Aim 1. Establish the roles of Pol IV vs. Pol V in cells; i.e., which does insertion vs. extension. Aim 2: Determine what amino acids in Pols IV and V control correct (dCTP) vs incorrect (dATP) insertion; e.g., why does Pol IV do correct (dCTP) insertion, while Pol V does incorrect (dATP) insertion, with [-ta]-B[a]P-N2-dG. Aim 3: Determine what lesion-bypass Pols are involved in G->A mutagenesis (dTTP insertion).
Crisp Terms/Key Words: enzyme activity, tissue /cell culture, bacterial protein, protein sequence, protein biosynthesis, DNA directed DNA polymerase, deoxyribonucleoside triphosphate, gene mutation, mutagen testing, mutagen, benzopyrene, adduct, Escherichia coli, aminoacid
DESCRIPTION (provided by applicant): Project Summary/Abstract We propose a CBPR study of the relationship between air pollution gradients and health effects in individuals living next to major highways. There is evidence that (1) people living close to highways experience significantly elevated exposures to constituents of motor vehicle exhaust including ultrafine particles (UFP; 0.01-1 microns) and black carbon; and (2) that motor vehicle pollution is associated with cardiac mortality and morbidity in adults, and asthma and reduced lung function in children. C-reactive protein (CRP), an inflammatory marker of risk for cardiac illness, has been shown to increase in response to changes in particulate exposure, making it a viable indicator of the potential impact on cardiac health. Our core study involves measuring 5 traffic-related pollutants (i.e., UFP, PM2.5, NOx, CO, black carbon, particulate PAH) in 6 neighborhoods within 400 meters of highways through in the Boston area. A background site >1000 m from highways will also be monitored. We will compete a scientific survey of residents living in the neighborhoods to determine pediatric asthma prevalence. We will determine the time residents spent within the near highway zone currently and rigorously measure highway pollution gradients in the neighborhoods. We will document exposures at work, school and while commuting. For a subset of non-smoking households we will obtain pulmonary function tests from children and analyze multiple blood samples per person from adults for CRP and fibrinogen. Our study will be (1) the first to test associations between highway pollution gradients and biological markers of health, (2) the first CBPR study of highway pollution, and (3) the most comprehensive collection of data on time spent in the exposure zone and many confounders and effect modifiers. We will conduct bivariate and regression analyses and have developed preliminary mathematical models that frame our approach to analyzing the large set of data. Our team consists of faculty at Tufts University and 6 co- investigators from community organizations that are concerned about the impact of highways on the health of residents in their communities. We will train and hire field staff from the communities and have an advisory board. We will link community participation to the science through participation in our steering committee and through our advisory board. Our study is designed to report useful information locally as well as influence pressing national policy needs. This proposal was developed to actively involve communities living very near to major highways in scientific research on the health effects of pollutant gradients coming off the highways (to about 200 meters). The study emphasizes the role of ultrafine particles on development of asthma in children, reduction of lung function in non-asthmatic children and increases in markers of inflammation and heart disease risk (C-reactive protein) in adults. PUBLIC HEALTH RELEVANCE: This proposal was developed to actively involve communities living very near to major highways in scientific research on the health effects of pollutant gradients coming off the highways (to about 200 meters). The study emphasizes the role of ultrafine particles on development of asthma in children, reduction of lung function in non-asthmatic children and increases in markers of inflammation and heart disease risk (C-reactive protein) in adults.
DESCRIPTION (provided by applicant): Social stressors, iron deficiency anemia, and lead exposure are all environmental risk factors for poor neurodevelopment in young children. All three factors are highly prevalent in poor communities, whether in Mexico or the U.S. There are scant human epidemiologic studies on chronic social stress and human neurodevelopment. In addition, previous studies of child development and neurotoxicants have typically analyzed covariate markers of social stress as independent predictors of neurodevelopment; and thus, only considered social factors as potential confounders in their data analysis. There are biological reasons to believe that stress potentiates the toxicity of iron deficiency anemia and lead poisoning. If so, then understanding this relationship may be the key to designing effective public health interventions designed to improve neurodevelopment and may explain the lack of effectiveness of prevention/treatment regimens for lead poisoning and iron deficiency. A prospective cohort study with a large sample size is needed to address the potential multiplicative effects of combined stress/iron deficiency anemia and combined stress/lead exposure in predicting infant development. This resubmission includes new data from a pilot study of stress-lead/stress-anemia interactions in 120 mother infant pairs conducted since the prior application, which demonstrates study feasibility and evidence for the hypothesized interactions. In this application we will measure maternal stress levels, infant iron deficiency anemia and lead exposure in a cohort of mother-infant pairs living in Mexico City and longitudinally measure maternal stress, iron stores, lead exposure, salivary cortisol and neurodevelopment beginning pre-natally and extending 3 years post-partum. This project will determine the independent effect of pre- and post-partum maternal stress in predicting infant development, and the multiplicative joint effects of maternal stress/iron deficiency and maternal stress/lead poisoning in predicting infant development.
Crisp Terms/Key Words: clinical research, neurotoxicology, women's health, psychological stressor, low socioeconomic status, neuropsychological test, early experience, prenatal stress, longitudinal human study, saliva, developmental neurobiology, lead poisoning, questionnaire, human subject, medical outreach /case finding, Mexico, preschool child (1-5), infant human (0-1 year), child psychology, blood test, microcytic /hypochromic anemia, circadian rhythm, cortisol
DESCRIPTION (provided by applicant): Polybrominated diphenyl ethers (PBDEs) are commonly used as fire retardants in consumer products such as electronics and furniture containing polyurethane foam. Structurally similar to polychlorinated biphenyls (PCBs), several PBDE congeners perturb hormonal signaling and are toxic to the nervous and reproductive systems of developing animals. Human body burdens have increased over the last several decades and are highest in the United States (US), where levels are more than an order of magnitude higher than in Asian and European populations in which PBDE levels are associated with decreased birth outcomes and increased risk of cryptorchidism. Diet and the indoor environment (dust, air) are likely important factors that contribute to personal exposure, but the predominant route of exposure is not known. Indeed, in a 2006 report on PBDEs, the US Environmental Protection Agency identified the "need for improved understanding of exposure to these chemicals and the associated hazards." We propose a comprehensive approach to examining human exposure to PBDEs. The proposed study has been designed to address a range of important gaps that exist in the PBDE research base and would represent the most complete assessment of human PBDE exposure to date. In a cohort of fifty Massachusetts residents, we will conduct three rounds of exposure sampling (at six month intervals) and examine three microenvironments: the home, workplace, and car. During each sampling round, we will concurrently assess PBDEs at multiple steps of the exposure-response continuum by (a) estimating the PBDE content of consumer products (potential sources) using a portable x-ray fluorescence analyzer, (b) measuring PBDEs in dust collected from the home, workplace and car (key microenvironments), (c) measuring PBDEs in personal air and hand-wipe samples (personal exposure), (d) measuring PBDEs in human serum (total absorbed dose), and (e) measuring hormone levels (potential early effects). This powerful study design will generate a rich dataset of repeated exposure measures and allow for a longitudinal assessment of PBDE body burden and hormone levels. Accordingly, the proposed research will provide a more complete understanding of how PBDEs enter the indoor environment, how the general population is exposed to PBDEs, which exposure routes contribute most to total absorbed dose, and the extent to which PBDE exposure may be associated with changes in hormone levels in humans. PUBLIC HEALTH RELEVANCE: The levels of polybrominated diphenyl ethers (PBDEs) found in the bodies of Americans have increased for decades and are an order of magnitude higher than in other countries where a few studies have found associations with adverse health outcomes. Although the indoor environment is hypothesized to be the major source of these flame retardants, the sources and routes of human exposure are not well understood. The proposed project will provide a more complete understanding of how PBDEs enter the indoor environment, how people are exposed, and the extent to which they may be associated with human hormones levels.
DESCRIPTION (provided by applicant): The aryl hydrocarbon receptor (AHR) is a ligand-activated transcription factor in the basic-helix-loophelix/Per-ARNT-Sim (bHLH-PAS) family. Halogenated aromatic hydrocarbons such as 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) act through the AHR to cause altered expression of xenobiotic-metabolizing enzymes and other changes leading to toxicity in mammals, fish, and other vertebrate animals. However, the normal functions of the AHR and the exact mechanisms by which TCDD and related chemicals act through the AHR to cause toxicity are poorly understood. We and others have recently characterized three members of the vertebrate AHR family: AHR1, AHR2, and AHR Repressor (AHRR), and we have found that there is additional AHR diversity in fish, with up to 5 genes per species. We propose an integrated set of studies in vertebrate model systems (fish, mouse cells, human cells) that will take advantage of the unique features of each model to better understand the function of AHR and AHRR proteins and their roles in dioxin toxicity and normal development. 1) We will use an RNA knock-down strategy employing morpholino anti-sense technology in zebrafish (Danio rerio) embryos to test the hypothesis that AHR1, AHR2, and AHRR have distinct roles during development and in dioxin toxicity. These studies will take advantage of the external development and transparent embryos of zebrafish. The effect of blocking AHR 1, AHR2, and AHRR expression on sensitivity to dioxin developmental toxicity and gene expression will also be determined. 2) We will test the hypothesis that multiple AHRs from fish have undergone subfunctionalization and can therefore be used to distinguish multiple functions of the human AHR. Multiple AHRs from medaka (Oryzias latipes) and pufferfish (Fugu rubripes) will be characterized in vitro and expressed in AHR-deficient mouse cells ¿ TCDD to determine whether individual fish AHRs regulate distinct subsets of genes controlled by the murine AHR. Taking advantage of the compact genome of pufferfish, we will also conduct in vivo mapping of AHR promoters and regulatory elements using zebrafish embryos expressing GFP reporter constructs. 3) We will characterize the function, expression, and regulation of the human AHRR, examining inducibility by AHR agonists, structure-activity and dose-response relationships, and regulatory elements involved in AHRR induction. We will test the hypotheses that AHRR inhibition of AHR function occurs through competition for ARNT and/or AHR response elements. These studies will provide a better understanding of the possible role of human AHRR in modulating dioxin effects.
DESCRIPTION (provided by applicant):
A new type of permeable, reactive composite is being developed for remediation and management of contaminated sediments. It consists of one or more filtering layers surrounding a reactive layer placed on the sediment surface. Hydrodynamic dispersion and pore fluid flow carry contaminants through the composite. The broad, long-term objective of this research is to develop a comprehensive understanding of: 1) how the gecicomposite would function most effectively in long-term field applications for different combinations of sediment types, sediment contaminants and pore fluid environments; and 2) how effectively the composite can reduce ecological and human health risks associated with sediment-bound contaminants. The specific aims of the research are to address the following fundamental issues: 1) What are the reactive materials that can be used in the composite that will most effectively immobilize or transform the target contaminants of PCB's, polyaromatic hydrocarbons (PAHs), and metals? 2) What is the relationship between the advective and diffusive components of contaminant transport for various soil-contaminant-pore fluid chemistry combinations? 3) How can results from bench-scale experiments that model the sediment-contaminant- composite system be used to predict the composite's scalability for field applications? 4) What are the effects of the physical presence of the composite and the composite-contaminant reactions on both the underlying sediment and overlying benthic organisms, in terms of supporting healthy and diverse benthic communities in these zones? 5) Does the geocomposite reduce the human health risks associated with contaminated sediments? The research will be performed in 4 phases: 1) sediment characterization for each Superfund site sediment to be tested, including physical, chemical, biological and consolidation testing; 2) batch chemical testing to assess and optimize reactivity between target contaminants and the composite reactive amendments to be testing in Phase 3; 3) bench scale tests to model the sediment-composite-benthic system using a computer automated device that combines sediment consolidation and contaminant transport through the sediment and composite, and assess the biological activity before and after sediment consolidation, as well as the biogeneration in the overlying 'clean' sediment; and 4) modeling of the process, accounting for seepage, consolidation, dispersion and reactivity due to adsorption and transformation. Relevance to Public Health: Contaminated sediments present a global public health challenge that requires new, innovative approaches. By immobilizing and/or neutralizing contaminants, the composite has the potential to minimize further contaminant migration and, in subaqueous environments, contaminant resuspension and diffusion into the overlying water column. This will limit the bioavailability of contaminants and bioaccumulation in aquatic food webs, which can be a pathway for human exposure via ingestion.
Crisp Terms/Key Words: biohazard detection, technology /technique development, water pollution control, environmental toxicology, soil sampling, soil pollution, high performance liquid chromatography, gas chromatography, sedimentation, scanning electron microscopy, mathematical model, model design /development, metal, disease /disorder prevention /control, halobiphenyl /halotriphenyl compound, geographic site, field study, waste treatment, environmental contamination, environmental health, disease /disorder proneness /risk, carbopolycyclic compound, computer simulation, chemical binding
In this revised application, funding is requested for the continued support of the NIEHS Center for
Environmental Health Sciences (CEHS) at the Massachusetts Institute of Technology (MIT). The overall focus of the MIT CEHS is to understand how toxic environmental agents perturb biological systems and to determine how such perturbations may affect human health. Founded in the early 1970' s, the CEHS underwent a renewal in leadership in 2001 and is now Directed by Leona Samson and Peter Dedon. As a result of this change in leadership and in response to reviewer criticisms, the Center has been completely reorganized. The current CEHS membership consists of 32 faculty members derived from a total of 8 MIT departments (in the Schools of Science and Engineering), plus three Departments in the Harvard School of Public Health (HSPH) and one in the Harvard Medical School (HMS). The MIT departments are Biology, Chemistry, Biological Engineering, Chemical Engineering, Civil & Environmental Engineering, Mechanical
Engineering, Nuclear Engineering, Electrical Engineering & Computer Science; the HSPH departments are Epidemiology, Nutrition and Environmental Health. The current Research Cores, each led by an MIT Professor, are (i) the Mutation and Cancer Research Core (led by Peter Dedon); (ii) the Bioengineering for Toxicology Research Core (led by Linda Griffith); and (iii) the Environmental Systems and Health Research Core (led by David Schauer). The current Facilities Cores are (i) the Bioanalytical Core (led by John Wishnok), (ii) the Genomics and Bioinformatics Core (led by Peter Sorger) and (iii) the Animal Models and Pathology Core (led by James Fox). The services available through the three Facilities Cores include sophisticated mass spectrometry, accelerator mass spectrometry, chromatography, transcriptional profiling and
computational analysis of such data, transgenic and knock out animal production, pathology services, and state-of the-art microscopy and imaging. Other units include the Administrative Core, the Pilot Project Program and the Community Outreach and Education Program (COEP). The Center also co-sponsors three departmental seminar series, and sponsors a CEHS member seminar series, an annual retreat and several other mechanisms designed to nurture interactions among CEHS members and to promote the activities of the CEHS and an awareness of environmental health science in the MIT community.
DESCRIPTION (provided by applicant) :
This amended application is designed to respond as completely as possible to concerns of the site visitors and of subsequent review groups. Accordingly, eight disciplinary Research Cores have been replaced by three multidisciplinary, integrated Research Cores. Facility Cores have been strengthened. The requirements and benefits of Center membership have been clarified. Leadership issues have been addressed. In toto, this program has been refocused and energized by these structural changes.
Our Center fosters active collaborations among three Research Cores: 1) Metals, 2) Urban and Occupational Particles, and 3) Organic Pollutants. The work of the Research Cores is greatly enhanced by three Facility Cores: 1) Biological Analyses, 2) Exposures and Environmental Analyses, and 3) Environmental Statistics. Finally, the investigators' ability to make their research and knowledge available to the public is greatly enhanced by the Community Outreach and Education Program (COEP). Other Center activities, such as pilot projects, new investigators, and program enrichment are catalytic mechanisms to achieve integration, interaction, productive, and innovative science. The objectives of this NIEHS Center are to generate new knowledge relating to the physiology, pharmacology, pathology, cell biology, molecular biology, genetics, and epidemiology of environmental disease, and to apply this knowledge to new modalities of prevention, diagnosis, and therapy. The investigators achieve these objectives through a variety of approaches, which range from studies of molecules and cells to those of whole animals and human populations. Through the organizational structure and financial support provided by the NIEHS Center Grant, the investigators' will increase the impact of their research and teaching in environmental health.
In toto, the Harvard NIEHS Center for Environmental Health continues to be a major focal point for environmental research and training in Boston. The NIEHS Center mechanism enhances connections and makes the Harvard NIEHS Center part of a national and international network.
DESCRIPTION (provided by applicant)
The goal of this application is to establish an NIEHS/NSF Center for Oceans and Human Health in Woods Hole Massachusetts. This will be a multi-institutional Center, with a mission to be a focal point for research and communication on issues at the intersection of oceanographic, biological and environmental health sciences, to improve the public health. The Center will involve investigators and facilities of the Woods Hole Oceanographic Institution, the Marine Biological Laboratory, and the Massachusetts Institute of Technology. The Woods Hole COHH will address fundamental issues of the distribution of biological agents with potential human health consequences, in the theme area of the temperate coastal ocean, including bays, harbors and estuaries. Within this geographic theme, Research Projects in the Center will focus on harmful algal blooms and on human pathogens and infectious agents, in both cases with focused and complementary physical oceanographic studies and biological studies of causative agents. The Center will comprise four Research Projects. The Research Projects will be coupled with an advanced Genomics Facility Core, establishing a combination of unusual strength, and supplemented by a Pilot Project program. The proposed Cores and Projects are: Project 1. Alexandrium Population Biology in the Gulf of Maine, Project 2. Hydrodynamic Forcing of Alexandrium Population Biology, Project 3. Human Pathogens and Coastal Ocean Processes, Project 4. Microecology and Evolution of Two Marine Pathogens. Genomics Facility Core.
Crisp Terms/Key Words: marine toxin, marine biology, saltwater environment, microorganism population study, public health, water microbiology
DESCRIPTION (provided by applicant)
The scientific theme for this competitive renewal continues the theme from the grant submission: effects of exposures to environmentally hazardous substances on reproduction and development in humans and wildlife. Special emphasis is placed on substances commonly encountered as a result of improperly managed waste disposal. The chemicals under study are organics, both halogenated and nonhalogenated compounds. The program will focus on the underlying mechanics of xenobiotics/ endocrine interactions and their effects to allow a better understanding of the implications of perturbations of reproductive and developmental processes by hazardous substances in the environment. Nine projects (5 biomedical and 4 nonbiomedical) will study: 1) Epidemiologic studies of neurodevelopment of a population exposed to perchlorethylene (PCE, a peroxisome proliferators) in drinking water, and epidemiologic techniques to study similar environmental problems (two biomedical projects); 2) receptor based mechanistic studies of the role of intracellular receptors and signaling pathways in the development of organisms and tissues (receptors/pathway: Ah receptor, AhR; peroxisome proliferator activated receptor, PPAR; estrogen receptor, ER; androgen receptor, AR; MAP kinase pathway) for important xenobiotics (planar halogenated aromatic hydrocarbons, PHAHs; polycyclic aromatic hydrocarbons, PAHs; and peroxisome proliferators, especially phthalates), and three biomedical projects; 3) Mechanisms of toxicity and resistance of fish populations to PHAHs and xenoestrogens involving receptors (AhRs, PPARs, and ERs) and cytochrome P450s (1 non-biomedical project and 2 biomedical); 4) studies of the mechanistic basis for reproductive and developmental effects on observed wildlife (including those mediated by AhR and ER) exposed to a complex mixture in surface water from a Superfund Site via groundwater and sediment (1 non-biomedical); 5) Mechanisms of oxidative dechlorination by an abiotic non-heme iron catalyst for remediation of a wide variety of xenobiotics, including all those under study in other projects.
DESCRIPTION (provided by applicant)
Particle air pollution has been associated with cardiovascular hospitalization and mortality in population-based studies throughout the world. The program project goal is to understand the relation of particle pollution to clinical, physiologic and biochemical endpoints reflecting potential mechanisms underlying these associations. The investigators have focused on endpoints influenced by autonomic regulation, and have found pollution effects on heart rate variability and on ventricular and supraventricular tachyarrhythmias. The investigators also have published or preliminary evidence for associations of particle pollution with elevated pulmonary/systemic inflammatory markers (e.g. exhaled nitric oxide (NO), intercellular adhesion molecule (ICAM-1), oxygen desaturation, ST-segment repression, elevated blood pressure, and, most recently, reduced vascular reactivity. Building on these observations, the principal investigators propose to broaden their focus to evaluate not only autonomic mechanisms, but also potentially interrelated vascular and inflammatory mechanisms for air pollution cardiovascular effects in humans. This evaluation requires a multidisciplinary approach joining experts in epidemiology, air pollution measurement, cardiology and electrophysiology; diabetes and measurement of vascular and inflammatory outcomes, and environmental statistics.
Projects proposed would provide data on a spectrum of at-risk subjects including the elderly, and populations with Type 2 diabetes, coronary artery disease, implanted cardioverter defibrillators, and acute cerebrovascular events. Primary outcomes of interest will include brachial artery diameter and endothelium-dependent flow-mediated dilation, heart rate variability, tachyarrhythmias in patients with implanted defibrillators, and acute ischemic cerebrovascular events. Markers of pulmonary and systemic inflammation and endothelial dysfunction will be evaluated as intermediate outcomes. The investigators will also differentiate effects of particle components closely associated with traffic (e.g., black carbon) and long-range transport (e.g., sulfate), with improved assessment of spatial as well as temporal differences in exposure. To carry out the proposed projects, several core services will be required. These will include the following: an Administrative Core; a Data Coordination and Statistics Core to apply consistent methodology to gathering, processing, checking and analysis of data; the Particle Exposure Characterization Core to provide detailed characterization and quantification of ambient pollution; and the Health Outcomes Core to provide vascular, electrophysiologic, and endocrine assessment and measures of inflammation/endothelial dysfunction. These studies will provide additional insight into the relative sensitivity of different subgroups to air pollution, and potential mechanisms whereby ambient pollution is associated with excess morbidity and mortality. Moreover, the study may provide clues as to how to develop preventive strategies to reduce this excess.
DESCRIPTION (provided by applicant): Chronic exposure to mercury, a highly reactive heavy metal, may increase cardiovascular disease (CVD) risk. Because the major environmental exposure to mercury is from fish intake, which may otherwise have significant cardiovascular benefits, and because adequate data are critical to ongoing policy decisions regarding mercury contaminants, elucidating the relationship between mercury and CVD risk is of great scientific and public health importance. However, results of prior studies have been inconsistent, and prior studies excluded women and did not evaluate stroke, a major cause of CVD morbidity and mortality. Selenium, an essential dietary trace element, plays an important role in antioxidant defense systems and may protect against both CVD and toxic effects of mercury. Such a protective effect would have direct implications for recommendations regarding optimal selenium intake and for assessing the potential impact of mercury contamination from fish intake in selenium-replete populations. However, the possible interaction between mercury, selenium, and cardiovascular risk is not well-established. The objectives of this application are to assess the associations between mercury and selenium exposure and risk of coronary heart disease (CHD) and stroke in women and men. It is hypothesized that: 1. Chronic mercury exposure is associated with higher risk of CVD (CHD and stroke) in women and men. 2. Selenium intake is associated with lower risk of CVD. 3. The relation between mercury levels and CVD risk varies depending on intake of selenium. These aims will be investigated using a prospective nested case-control design among women and men participating in two large U.S. cohort studies, including 6,470 incident CVD cases. Exposures will be ascertained using stored toenail clippings, a reliable biomarker of chronic mercury and selenium intake. This study provides a unique and cost-efficient opportunity to evaluate the importance of mercury and selenium intake for cardiovascular risk in women and men. This research will greatly clarify the potential risks and benefits offish intake, mercury exposure, and selenium intake for cardiovascular disease prevention, fulfilling the NIH's mission to pursue fundamental knowledge about human health and illness and the application of that knowledge to extend healthy life and reduce burdens of death and disability.
DESCRIPTION (provided by applicant): Large population studies of fine particulate air pollution have shown a link with heart disease, but for practical reasons they have not included direct assessment of biochemical risk factors or personal exposures. In this Exploratory/Developmental Research application we develop the methodology to collect blood samples and recruit study participants from a national cohort of unionized trucking company workers who have regular occupational exposures to fine particulate matter (PM) from motor vehicle exhaust. With the cooperation of the Teamsters we propose to assess the rate that unionized trucking company workers agree to provide a blood sample and complete a health questionnaire for a health study. Phlebotomy and study recruitment sites will be located in Teamster union halls near large trucking terminals in Chicago, IL and Carlisle, PA that employ 3,481 workers. The demographics and exposure distribution of subjects who provide a blood sample will be compared to workers who do not provide a blood sample. This strategy of locating phlebotomy centers in Union halls will be compared to the results from a companion study where another group of Teamsters are recruited by mail and requested to provide a blood sample by overnight mail using a blood collection kit. Preliminary information regarding the relationship between combustion particle exposures (based on job title and terminal location) and blood markers of inflammation that include C-reactive protein (CRP, measured using a high sensitivity assay), fibrinogen, interleukin-6 (IL-6), and markers of vascular endothelial activation and inflammation (soluble intercellular adhesion molecule-1 (slCAM-1), and soluble vascular cell adhesion molecule-1 (sVCAM-1)) will be obtained. Cross-sectional relationships between combustion particle exposures, blood markers of inflammation, and concentrations of 1,2-naphthoquinone and 1,4- naphthoquinone albumin adducts, specific biomarkers of polycyclic aromatic hydrocarbons (PAHs) will be assessed. This approach will offer the possibility of prospective personal monitoring for a large population, greatly increasing the power to detect health effects and link them to personal risk factors and exposure to motor vehicle exhaust. The results from the proposed exploratory and developmental study will aid us in designing a prospective study to determine the relationship between PM exposures from motor vehicles and cardiovascular disease. This is of public health importance because large segments of the general US population have exposures to fine particles similar to trucking company worker exposures.
Crisp Terms/Key Words: air pollution, engine exhaust, biomarker, blood toxicology, vascular endothelium, blood chemistry, carbopolycyclic compound, inflammation, medical outreach /case finding, human subject, data collection methodology /evaluation, naphthoquinone, occupational hazard, particle counter, particle, human population study, environmental toxicology, method development, clinical research, sample collection, environmental exposure, cardiovascular disorder risk
DESCRIPTION (provided by applicant): Since the late 1980s, studies of acute and chronic exposure to particulate air pollution have shown it to be associated with early death. It was quickly apparent that most of these early deaths were from cardiovascular disease. Definition of the underlying mechanisms that may explain the epidemiological findings of cardiac mortality/morbidity associated with exposure to ambient particles, definition of subpopulation at increased risk of adverse health outcomes from particulate matter, and understanding the differential toxicity of different types of particles are among the research priorities defined by the U. S. National Research Council. We propose to examine the association of different types and sources of particle exposure with cardiovascular and pulmonary responses along a mechanistic pathway, using data from the Normative Aging Study, a prospective cohort study conducted by the Veterans Administration in the greater Boston area. Specifically, we will examine whether short term changes in automatic function, as indexed by heart rate variability, are associated with particle exposure, and determine whether that effect is mediated by part by oxidative stress and metal toxicity by examining effect modification by genotypes that modify defenses against oxidative stress and metal handling. We will similarly examine associations, and effect modification, for vascular function, as indexed by a non-invasive pulse Doppler measures of arterial compliance and blood pressure, and examine a direct association with systemic markers of inflammation and oxidative stress (c reactive protein, Sicam, Svcam, homocysteine, and 8-hydroxyguanosine.
DESCRIPTION (provided by applicant): Particulate air pollution is an important public health problem with respect to cardiovascular morbidity and mortality. Important strides have been made in understanding the neural and cardiac mechanisms involved. Heart rate variability analysis revealed important disturbances in autonomic tone during exposure to concentrated air particles (CAPs). We have obtained evidence in our canine models that CAPs may increase the severity of ischemia during acute coronary artery occlusion. These findings provide the direction for our studies, which are intended to expand our understanding of the mechanisms of air particulate exposure on the cardiovascular system. The specific aims are: 1) To determine the effect of ambient air particles on coronary hemodynamic function and arterial blood pressure in conscious dogs using CAPs exposures. 2) To determine whether the influence of ambient air particles on coronary artery blood flow and resistance is exacerbated by coronary artery stenosis in conscious dogs using CAPs exposures. Our studies will continue to employ the Harvard Ambient Particulate Concentrator (HAPC), a device that can increase ambient particle concentrations up to 30x without changing the physical or chemical characteristics of the particles; 2) a typical urban aerosol; 3) animal models of disease including myocardial ischemia in dogs to simulate the condition of compromised humans with ischemic heart disease, the primary substrate for adult cardiac mortality. The animals will be chronically instrumented with flow probes and telemetry devices to monitor arterial blood pressure and EKG. Coronary blood flow and pressure will be monitored in normal dogs and in dogs with partial coronary artery stenosis and compared during sympathetic or parasympathetic nerve blockade. The vasoactive balance controlled by endothelins and nitric oxide will be assessed during regulation of the coronary vascular response. The effects of components of pollutants will be systematically studied. Thus, the present application will entail unique application of comprehensive techniques to improve understanding of the mechanisms whereby CAPs can exert its deleterious influences on the heart and circulation
DESCRIPTION (provided by applicant): Population-based epidemiologic studies of communities in the United States have revealed a consistent association between ambient particulate air pollution and increases in morbidity and mortality. The observed increases result from both respiratory and cardiovascular diseases. Similar associations have been observed for rates of hospital admissions for respiratory and cardiovascular diseases for subjects over age 65. Results from the first phase of this study (ES R01 09860) showed significant cardiac and airway changes after both environmental and occupational particulate exposures. The objective of this application is to investigate the role of both occupational and non-occupational exposure to particulates in the development of respiratory and cardiac responses in boilermakers. We will employ a detailed, continuous exposure assessment to PM2.5 with and PM1.0 with repeated measures of biologic and physiologic markers of response. In addition, we will employ novel techniques for the assessment of particulate-induced physiologic responses, including gene expression using mRNA microarrays. Hypotheses to be addressed in this established cohort of boilermakers include: (1) Short-term exposure to particulates from occupational, ambient, and indoor exposures results in airway inflammation and obstruction measured by serial expired NO and FEV1; (2) Chronic exposures to particulates result in long-term decreases in expired NO and FEV1; (3) Exposure to particulates results in acute changes in cardiovascular function measured as heart-rate variability, heart-rate, and blood pressure; (4) Exposure to particulates results in increased acute phase reactant (c-reactive protein, fibrinogen) concentration in the serum; (5) Exposure to particulates induces systemic responses resulting in alterations in white blood cell gene expression assessed by mRNA microarray analysis pre- and post-exposure; (6) Common chronic medical conditions such as hypertension, COPD, and asthma predispose particulate-exposed individuals to changes in cardiac function (heart rate, heart rate variability, and blood pressure). The results of this work will have important implications for preventive efforts aimed at reducing particle-associated morbidity and mortality.
DESCRIPTION (provided by applicant): Although the use of polychlorinated biphenyls (PCBs) and many of the chlorinated pesticides, including DDT, are now prohibited or restricted, there remains concern over their potential human health effects because they persist in the environment, concentrate up the food chain, and are stored in adipose tissue. Laboratory studies show associations of environmentally relevant levels of PCBs and chlorinated pesticides with infertility and pregnancy loss through mechanisms that disrupt embryo development. Epidemiologic studies report associations of PCBs and DDE (stable DDT metabolite) with pregnancy loss. In the proposed study, we will explore the developmental toxicity of chlorinated compounds in women undergoing in vitro fertilization (IVF), which can be used as a model to assess early development, normally unobservable. Developmental endpoints that will be measured include oocyte production, fertilization, and pre- and post-implantation embryo development. Failure at these developmental stages results in infertility and pregnancy loss. Other measured endpoints include clinical pregnancy loss (spontaneous miscarriage and stillbirth) and peri-natal outcomes, such as birth weight. The proposed study is cost-effective because it uses data and specimens from a recently completed NICHD funded multi-center study on epidemiologic predictors of IVF success. In the multi-center study there is data on 2494 couples undergoing 5071 IVF cycles between 1994 and 2003. Serum and ovarian follicular fluid were collected and archived from the female partner and will be used to measure PCBs and chlorinated pesticides such as DDE, hexachlorobenzene, mirex, and the chlordane metabolites, trans-nonachlor and oxychlordane. Subjects completed detailed health and lifestyle questionnaires and information on IVF protocols and pregnancy outcomes were abstracted from patient medical records. The study design is novel because it uses IVF as a model system to study the associations of PCBs and chlorinated pesticides with failure of early development, which manifests clinically as infertility and early pregnancy loss. Early developmental failure represents the majority of losses of pregnancy and is considered the most sensitive developmental stage to PCB and chlorinated pesticide exposure. Since the etiology of infertility and early pregnancy loss remains largely unexplained, the identification of potential environmental risk factors will have large public health significance.
DESCRIPTION (provided by applicant): Increased malformations of the genital tract and hormone-related cancers are significant problems in the industrialized world. Suspicions have focused on environmental estrogens as one causal agent. Among them, bisphenol A (BPA) is of particular interest due to widespread human exposure. Perinatal exposure of rodents to low, environmentally-relevant doses of BPA induces pleiotrophic effects in estrogen target tissues that manifest long after exposure has ended. In particular, altered sexual differentiation of a nucleus important for estrous cyclicity, and altered gonadotropin-releasing hormone (GnRH) neuronal activation may have repercussions on fertility and fecundity, while altered morphogenesis of the mammary gland may impair lactation. We expect that effects observed in estrogen target tissues of BPA exposed females will impair their ability to produce viable' and healthy offspring. The proposed studies will establish a causal mechanistic chain for BPA action encompassing cellular, tissue and organismal levels of organization; hence, they will be both integrative and analytical. On the integrative side, we will evaluate the reproductive success of perinatally exposed female mice. This information is essential to elucidate the physiological consequences of the molecular events described in the previous funding cycle. On the analytical side, we propose a dual approach to study how BPA alters the tissue organization of two important target tissues, the developing hypothalamus (HYP) and the mammary gland (MG). The HYP is critical to overall reproductive success, and the MG is critical to the survival of the neonates. In addition, the HYP can influence MG development by modulating pituitary gonadotropins and ovarian hormone synthesis and prolactin release. Aim 1: How does BPA affect the reproductive outcome of perinatally exposed females? Fertility, fecundity, and MG function will be assessed in order to define the reproductive impact of developmental low dose BPA exposure. Aim 2: How does BPA exposure alter tissue organization in the developing HYP? We hypothesize that BPA alters the architecture and connectivity of nuclei important for the regulation of gonadotropin release. We will examine these nuclei for: i) changes in patterns of cell survival, apoptosis, and connectivity; ii) expression of steroid receptors, enzymes of testosterone metabolism, and factors downstream of estrogen action such as glutamic acid decarboxylase and astrocyte differentiation. Completion of these studies will identify mechanisms underlying altered GnRH neuronal activation. Aim 3: How does BPA exposure affect gene expression and tissue organization in the MG? We hypothesize that: i) BPA acts as a morphogen directly on the MG anlagen (to be tested by QRT-PCR in MG organ culture); ii) BPA effects are mediated by ER alpha and/or beta (to be tested by QRT-PCR using null ER mice), and iii) these initial events translate into altered stroma-epithelium interactions. To dissect the effects resulting from BPA exposure of the MG anlagen from systemic effects due to the action of BPA on the endocrine system, the MG of BPA exposed and unexposed animals will be transplanted into exposed and unexposed hosts. To assess whether the stroma, the epithelium or both compartments are permanently altered by BPA exposure, tissue recombination studies will be performed. This Aim will begin to reveal the mechanisms by which BPA disturbs the organization and architecture of an estrogen target organ. The realization of this project will provide mechanistic information linking BPA action in target tissues and its organismal consequences. It will also reveal whether current levels of environmental exposure produce significant health effects in a surrogate model. This information is critically needed to develop public policy on endocrine disruption.
DESCRIPTION (provided by applicant): The goal of this application, entitled "Gastroenteritis and Extreme Weather Events in Elderly (GEWEL)" is to examine how temporal and spatial patterns of gastrointestinal diseases (GID) respond to extreme weather events at the national and regional levels. We postulate that variability in the GID incidence in the elderly is sensitive to a change in meteorological parameters. We assume that frequency and intensity of extreme weather events, such as heat waves, droughts, heavy rainfalls and floods, can alter the incidence of food borne and waterborne infections. We expect that the effect of extreme events on health will depend on vulnerability of the affected population and the geographical area. In the proposed study we will: 1) examine the effect of extreme temperature and precipitation on severity and composition of food borne and waterborne infections among the elderly in Massachusetts using -50,000 laboratory-confirmed cases and approximately 100,000 hospitalization records of six enteric diseases (1992-2004); 2) assess the impact of extreme weather events on hospitalizations for acute GID in the US elderly across climatic zones using nation-wide dataset of approximately 2,500,000 hospitalization records (1984-2004); and 3) develop, test and implement analytical models for assessing long-term, intermediate, and short-term effects of an extreme event on GID. A multidisciplinary team is fully capable of completing an integrated project that will explore a complex system of health and environmental interactions and develop novel methodology which will have important implications for environmental and health policy. Reliable, objective and accurate estimates of incidence of enteric infection related to environmental stressors, better understanding of mechanisms of human environmental interactions, and the impact of socio-economic and demographic factors on such interactions are crucial for the development of preventive strategies to reduce morbidity and mortality and to improve the quality of life among the elderly. Based on gained knowledge, reliable and efficient warning systems can be built for enteric infections, including emerging pathogens; the most vulnerable geographic areas can be identified; and surveillance systems can be substantially improved nation-wide.
Crisp Terms/Key Words: geographic difference, geriatrics, interdisciplinary collaboration, clinical research, human data, water pollution, temperature, environmental stressor, socioeconomics, longitudinal human study, precipitation, mathematical model, model design /development, hospital utilization, gastrointestinal infection, gastroenteritis, gastrointestinal disorder, food contamination, environmental health, atmosphere /weather, disease /disorder proneness /risk, enteric bacteria, human old age (65+)
DESCRIPTION (provided by applicant): The prevalence of obesity has increased several-fold over the last few decades in children, adolescents, and adults. Some evidence suggests that there has been a disproportionate increase in the level of central adiposity, which carries an even greater disease risk than peripheral fat. Changes in diet and exercise habits over the past few decades are undoubtedly linked to the current epidemic of overweight and obesity, and changes in the quality and composition of the diet may also play a role. Recently, interest has emerged in the possibility that environmental chemical contaminants may also contribute to the growing problem of obesity. Limited evidence from laboratory studies suggests that synthetic chemicals may affect obesity- related pathways by changing hormone levels or altering gene expression, but virtually no human studies have been conducted. In this proposed epidemiologic study, cross-sectional data on a sample of approximately 5000 participants, aged 6 years and older, from the 1999-2002 National Health and Nutrition Examination Survey (NHANES) will be analyzed to determine whether blood and urine levels of environmental contaminants are associated with level of body fat. The measures of body fat to be examined include body mass index, waist circumference, subscapular skin folds, and the amount and type of body fat, estimated using dual-energy x-ray absorptiometry (DXA). The study will examine the potential adverse effects of exposure to phthalates and heavy metals as well as the possible protective effects of phytoestrogens such as genestein on total body fat and the level of central adiposity. Potentially important interactions between these environmental exposures and age, gender, various dietary factors, activity levels will also be examined. Finally, the study includes a secondary analysis that takes advantage of the repeated NHANES surveys over four decades to answer the question of whether the level of central adiposity has increased over time in separate age, race, and gender subgroups, after adjusting for increases in BMI. This study is designed to use existing data to explore the novel hypothesis that environmental contaminants may, in part, be responsible for the increasing rates of total obesity and the increases in visceral fat deposition in children and adults. The strengths of the study include its large sample size, state-of-the-art measurements of environmental contaminants from blood and urine, precise measurements of BMI, DXA-derived measures of central adiposity, and extensive information on potential confounders and effect modifiers. The study is limited by its cross-sectional nature. Nevertheless, it may provide important clues for future, epidemiologic studies of the effect of these environmental contaminants on the risk of obesity.
Crisp Terms/Key Words: endocrine disrupting compound, anthropometry, weight gain, environmental exposure, phytoestrogen, human data, age difference, photon absorptiometry, racial /ethnic difference, gender difference, epidemiology, phthalate, dietary fiber, nutrient intake activity, heavy metal, statistics /biometry, questionnaire, body physical activity, environmental contamination, disease /disorder proneness /risk, adipose tissue, urinalysis, blood chemistry, abdomen, obesity, body composition
DESCRIPTION (provided by applicant): A temporal trend towards an earlier age of onset of puberty has been reported in both girls and boys. Although specific etiologies have not been clearly identified for this reported change in the age of pubertal onset, the effects of environmental chemicals as well as that of overnutrition and obesity on the reproductive axis have both been implicated as potential factors. Environmental chemicals of concern are a family of compounds that includes polychlorinated dibenzo-p-dioxins, dibenzofurans and polychlorinated biphenyls. These compounds are reproductive and developmental toxins in laboratory animals and in limited human studies. In the proposed application, we target a population in Chapaevsk, Russia, an industrial town that has high documented environmental levels of dioxins and PCBs. We propose to prospectively extend follow-up of our cohort of 550 eight- to nine-year old boys, in which we are investigating the association of dioxins and PCBs with: 1) the timing and tempo of pubertal development; 2) linear growth, weight gain, body composition (per cent body fat), and body mass index; and 3) biochemical changes in hormones that regulate growth and pubertal development. At recruitment, baseline examinations are performed that include physical and physiologic markers of growth and pubertal development. Physical markers include anthropomorphic measurements and determination of pubertal maturation by Tanner staging for genital and pubic hair development and assessment of testicular volume. Physiological markers consist of biochemical measures of the hypothalamic-pituitary-gonadal axis, assessed by serum gonadotropins (follicle-stimulating hormone and luteinizing hormone), sex steroids, inhibin-B and Mullerian inhibiting substance, as well as hormones that regulate somatic growth and metabolism, such as thyroxine, thyroid stimulating hormone, leptin, and insulin- like growth factor-1. Blood samples are collected from the boys and their mothers for dioxin and PCB analysis. Chemicals that alter growth and pubertal development have significant clinical and public health implications for both childhood and adult health. Specific implications of altered growth and development include impaired fertility, increased risk of obesity, and risk of behavioral and psychological disorders. The proposed study is unique since it will represent one of the first large longitudinal studies to explore the relationship of dioxins and PCBs with growth and development in pre-pubertal boys.
Crisp Terms/Key Words: endocrine disrupting compound, environmental exposure, disease /disorder etiology, environmental toxicology, male, precocious puberty, reproductive development, embryo /fetus toxicology, longitudinal human study, pituitary gonadal axis, dioxin, questionnaire, human subject, case history, halobiphenyl /halotriphenyl compound, hormone regulation /control mechanism, middle childhood (6-11), human puberty, child physical development, urinalysis, blood chemistry, weight gain, clinical research, health behavior, hypothalamic pituitary axis, Commonwealth of Independent States, body physical characteristic
DESCRIPTION (provided by applicant):
Breast cancer is now the second most frequent cause of cancer death among women in the United States. Environmental chemical exposure and bioaccumulation throughout a women's lifetime are thought to play a substantial role in the rise in breast cancer incidence. The overarching hypothesis in our original application was that environmental carcinogens, which induce genetic and epigenetic events, set in motion interacting signaling cascades that promote a multistep process leading to mammary tumor formation. Three gene systems were the focus: the aromatic hydrocarbon receptor (AhR), the CK2 and GSK3 kinases, and the NF- ?B family of transcription factors. This paradigm-shifting model has been confirmed, and a new hypothesis on the role of these critical regulatory pathways in tumorigenesis has been developed. The investigators now propose that the interactions among these three systems play key roles at several stages of the multistep signaling process that leads to altered cell morphology and invasive tumor formation. The individual hypotheses to be tested are: Project 1: As mammary epithelial cells progress from normal to immortalized cells and then to invasive tumors, AhR activity is modified through interactions with environmental chemicals, other transcription factors and cofactors to differentially regulate target gene transcription and to effect changes in cell growth and invasiveness. New information will be obtained on AhR function in normal as compared with malignant cells, and on the molecular and functional outcomes, particularly with regard to tumor invasiveness, of constitutively active as compared with environmental chemical-activated AhR. Project 2: Upregulation of CK2 and the Wnt pathway, resulting from environmental carcinogen exposure, contributes to epithelial to mesenchymal transition (EMT), which promotes tumor cell invasion and metastasis. Targets of CK2 in this process will be identified, and the mechanism by which 7,12-dimethylbenz(a)anthracene (DMBA) interacts with CK2 in tumorigenesis and promotes CK2 upregulation will be studied. Project 3: Genetic and epigenetic events initiated by environmental carcinogen exposure induce or enhance the activity of multiple NF-?B complexes, thereby promoting a more invasive phenotype of breast cancer. In particular, studies will focus on elucidating the roles of c-Rel, RelB, and IKKe/i kinase, and tumor inhibitory effects of green tea polyphenols. Thus, this application focuses on understanding signal transduction pathways impacted by environmental carcinogen exposure, which promote neoplastic transformation, with particular emphasis on elucidating the mechanisms that influence development of invasive breast disease. The investigators will employ innovative technologies, novel in vitro model systems, transgenic mice generated in the P01, and common cell lines and reagents. These studies will provide important information on the potential link between environmental factors and critical signaling pathways in the pathogenesis of breast cancer invasion and on the targeting of these interacting signaling pathways with novel prevention or treatment modalities.
BACKGROUND
DESCRIPTION (provided by applicant): Obesity is one of the most common health problems in the US. The rapid increase in the prevalence of obesity in the U.S. suggests that primary causes may be environmental. Although changes in diet and physical activity have long been considered the major culprit in this regard, there is novel information that points to environmental endocrine disrupters as potential causative agents.
While performing experiments aimed at understanding the effects of perinatal exposure to environmentally relevant levels of the estrogen bisphenol-A (BPA) we observed a substantial increase in body weight that persisted throughout the adult life of these animals. Further exploration revealed adiposity, hyperphagia and glucosuria. The nature of the metabolic aberrations underlying this type of obesity is unknown, but its persistence long after cessation of exposure suggests that BPA exerts early, irreversible effects on metabolic programming. Our working hypothesis is that exposure to BPA during fetal and neonatal life disrupts the development of key metabolic regulatory networks (in systemic organs and/or in the CNS), leading to sustained alterations in expression of genes involved in regulation of food intake, triglyceride anabolism and catabolism, and/or glucose utilization.
Specific Aim 1 will assess the nature of the weight gain in the offspring of pregnant mice exposed to BPA from gestational day 9 through lactation. The following parameters will be measured postnatally and at two later ages: body size; mass and linear growth rates and composition, food consumption, tissue mass and adipose cell number, size of individual adipose depots; fasting levels of metabolically relevant plasma metabolites and hormones.
Specific Aim 2 will explore the expression pattern of genes encoding metabolically relevant genes as indicators of the relative activity levels and state of regulation of major anabolic and catabolic pathways in tissues involved in the control of energy balance. They include C/EBP-alpha, PPAR-gamma, FAS, Glut-4, HSL, and leptin in white adipose tissue, UCP1 in brown adipose tissue, and POMC and NPY in the hypothalamus. These parameters will be investigated using real-time RT-PCR; mRNA levels will be measured during the period of BPA exposure, and at two later points at the same ages and doses examined in Aim 1.
These studies will indicate the extent of obesity and whether there is differential involvement of specific adipose depots or increases in adipocyte numbers following perinatal BPA treatment. They also will provide indicators of the major candidate pathways involved, and will guide the development of hypotheses about the mechanisms underlying this phenomenon, including dysregulation and interactions of CNS and peripheral pathways. It is expected that these results, in turn, may generate a better understanding of the obesity problem, and provide elements to help devise a sound policy addressing this serious public health problem.
Crisp Terms/Key Words: weight gain, leptin, environmental toxicology, lactation, early experience, perinatal, embryo /fetus toxicology, phenol, estrogen analog, hormone regulation /control mechanism, brown fat, adipose tissue, obesity, bioenergetics, laboratory mouse
DESCRIPTION (provided by applicant): Polychlorinated biphenyls (PCBs) contaminate seafood and freshwater fish worldwide. Current risk assessment is based on immunotoxicity in rodents, although they differ from humans in regard to immune system development; epidemiological confirmation is lacking. We have carried out a pilot study in the Faroe Islands and found that prenatal exposure to dioxin-related PCB congeners appears to affect the antibody response to childhood vaccinations, possibly mediated via a toxic effect on prenatal T-cell programming. This North Atlantic fishing community is unique and highly suitable for prospective population-based studies. Average PCB exposures vary more than 100-fold within the population, and confounding variables other than marine food contaminants are of limited concern. We now propose to examine a birth cohort of 547 children born over a 2-year period from 1 April 1998 at the Faroes. The children will first be examined just before the final vaccination booster against tetanus and diphtheria at age 5 years. An additional blood sample will be obtained 4-8 weeks later. The children will again be examined at approximately age 7 years. Outcome variables will include antibody responses to vaccines, acute-phase reactants, and questionnaire information on childhood infections. Stored maternal serum from late pregnancy, and serum from the children will be analyzed for PCBs and other PHAH contaminants, and we will use a novel method to assess the in vitro activation of the arylhydrocarbon receptor (AhR or 'dioxin' receptor) by the contaminants present in serum. Mercury will be measured in cord blood, postnatal blood and hair. Statistical analyses will include multiple regression analysis, supplemented by mixed model analyses, structural equation models, and determination of benchmark doses of individual contaminants, their mixtures, and integrated exposure measures. These results are expected to extend substantially the current documentation for PCB risk assessment.
DESCRIPTION (provided by PI): The goal of these studies is to determine the molecular mechanisms by which environmental chemicals impair immune cell development. While many studies demonstrate that environmentally ubiquitous polycyclic aromatic hydrocarbons (PAH) suppress the mature immune system, relatively few have evaluated their effects on the developing immune system. To bridge this gap, we exploited B cell development models to define how PAH affect bone marrow B cells. We demonstrated that PAH suppress B lymphopoiesis by inducing pro-B and pre-B cell apoptosis mediated indirectly through stromal cells constituting the hematopoietic microenvironment. These studies led to the hypothesis that the developing B cell compartment is exquisitely sensitive to environmental chemicals, which inappropriately activate B cell death programs.
Our recent studies have begun to elucidate the cascade of signals that executes the B cell death program. The working model built on these studies proposes at least two potentially overlapping pathways, one involving caspase-8 and another involving mitochondrial activation. A combination of unique resources, including apoptosis gene-defective, nontransformed early B cell lines and a stable of knockout mice will be exploited to test and to refine this model. Three specific aims are proposed: 1) Define PAH-induced, caspase-8-dependent, apoptosis signaling pathways in bone marrow B cells (D. 1): We will: a) determine the role of death receptors in caspase-8 activation, b) evaluate the contribution of JNK, PKR, and ASK1 kinases to caspase-8-dependent apoptosis, and c) identify components of activated caspase-8 complexes by mass spectrometry (MALDI-TOF). 2) Evaluate factors contributing to mitochondria-dependent apoptosis signaling. (D.2): The elements of a PAH-induced mitochondrial apoptosis amplification pathway will be mapped out. Specifically, we will: a) determine the roles of caspases-2 and -8 in mitochondrial activation mediated by Bid, b) assess the contribution of JNK, PKR and p53 in caspase-independent mitochondrial activation, and c) define where NF-kappaB down-regulation, a prerequisite to PAH-induced pro/pre-B cell death, interfaces with caspase signaling. 3) Investigate in vivo correlates of the apoptosis pathway mapped out in vitro. (D.3): To build on results obtained in vitro, we will use gene-knockout mice: a) to determine the relative sensitivities of bone marrow B cell subsets to AhR-dependent apoptosis signaling in vivo and b) to investigate PAH-induced, caspase-dependent death signaling in bone marrow B cell subsets in vivo. These studies will not only contribute greatly to our understanding of the molecular effects of PAH on developing B cells, but they also will validate a platform that is easily applicable to the study of other immunotoxic environmental chemicals and other hematopoietic cell subsets on the molecular level.
DESCRIPTION (provided by applicant): Most carcinogens form covalent products, or adducts, with DNA. Adducts are believed to drive the genetic changes that convert normal cells into cancer cells, which then outgrow into a tumor. Factors that influence the formation or removal of adducts, therefore, are likely to be important determinants of human susceptibility to carcinogenesis. Moreover, agents that damage DNA can modify targets not directly relevant to cancer; directly or indirectly, DNA damaging agents may play important roles in initiating or promoting a host of other diseases. The work described below is an effort to understand how DNA adduction integrates with other biochemical factors, determinable by modern analytical tools, to define the differences in sensitivity to environmental agents that are associated with age and gender. The main focus of the work deals with aflatoxin B1 (AFB1), an important human liver carcinogen that is associated with most cases of hepatocellular carcinoma, especially when toxin works in concert with hepatitis viruses. Our work will address four gaps in knowledge. First, we shall provide a high resolution map of the biological networks of both genders of the B6C3F1 mouse at specific time points from fetus through infancy and adulthood and determine how those networks respond to AFB1. Second, the gene network data we produce will be anchored to sensitive detection of DNA adducts, using a tool that will even detect adducts in the fetus of an exposed mother. Accelerator Mass Spectrometry (AMS) will be used to examine the formation and fate of DNA adducts at sensitive and resistant stages of life. Third, we shall administer to mice a chemo-interventive agent, sulphoraphane, that we expect will alter metabolic networks in a manner that will protect pre- born, infant and adult animals from this environmental insult. Finally, our experiments with AFB1 will be coupled with a more limited investigation of the genotoxic effects of four compounds from the NTP data set. These additional agents include4-aminobiphenyl (ABP), 2-amino-1-methyl-6- phenylimidazo[4,5-b]pyridine (PhIP), acrylamide and 17¿-estradiol (E2). An important goal of this research is the development of a host of new biomarkers that can be applied to the mouse model, and later other models that are used to predict the impact of environmental agents on humans.
DESCRIPTION (provided by applicant): In order to contend with ongoing physiological stress imposed by the local microenvironment or by endogenous oxidative processes, cells have evolved a complex network of signaling molecules to detect these perturbations and to recruit an ensemble of responses to countermand the insult. These afferent signals converge on a host of transcription factors that coordinately regulate the expression of genes which give cells pause to rectify the cellular injury or to execute an apoptotic program. Arguably, the most ubiquitous source of environmental stress for terrestrial organisms is ultraviolet radiation (UVR) and the most lethal consequence of this widespread exposure in human is malignant melanoma. The success of the aspiring melanoma cell is critically dependent on its ability to negotiate the physiological challenges of early UVR threat. In order to better understand these early events, we recently set out to identify UVR-inducible genes in primary pigment cells. In preliminary studies, we discovered that the receptor tyrosine kinase, EPHA2, is upregulated by UVR in melanocytes and other cells types and by a common melanoma oncogene, activated NRAS. Most notably, EphA2 appears to be an essential mediator of apoptosis in response to UVR - the most common exogenous stressor for these cells. In this application, our overarching goal is to understand the role of EphA2 in regulating the UVR stress response and potential downstream tumorigenesis. In the first two Aims, we will explore the mechanism by which EphA2 induces apoptosis and expand on its position as a UVR stress-inducible gene in intact skin. In the last Aim, we will investigate the link between UVR stress and oncogenesis in a novel murine model of melanoma photocarcinogenesis.
PROJECT NARRATIVE. Malignant melanoma is possibly the most lethal consequence of long-term ultraviolet radiation (UVR) stress and mutagenesis. In preliminary studies, we identified a UVR stress-inducible gene, EPHA2, which empirically behaves as an essential mediator of UVR-induced apoptosis. As this gene has been implicated in the pathophysiology of multiple cancers, we propose to elucidate the mechanism by which EPHA2 modulates cellular death or survival. These clues will undoubtedly drive therapeutic developments in metastatic melanoma, which, to this day, remains incurable.
DESCRIPTION (provided by investigator): There is uncertainty about the effect of low-level methylmercury (MeHg) exposure on neurodevelopment. Two seminal studies conducted in the Faroe Islands and the Seychelles Islands of prenatal MeHg exposure report inconsistent conclusions with respect to cognitive and behavioral outcomes. The primary aims of the proposed research are to investigate: 1) the association between prenatal MeHg and attention-associated behaviors among infants using the Neonatal Behavioral Assessment Scale (NBAS); and 2) the association between MeHg and behaviors associated with Attention Deficit Hyperactivity Disorder (ADHD) among school-aged children (median=8 years) using the Conners' Rating Scale for Teachers (CRS-T), the Neurobehavioral Evaluation System 2 (NES2) Continuous Performance Test (CPT), and components of the Wechsler Intelligence Scale for Children-Third Edition (WISC-III). We will also investigate a potential synergistic effect of MeHg and polychlorinated biphenyls (PCBs) on behavior and evaluate gender differences for the MeHg and behavior associations. Finally, we will assess the potential confounding effect of fish consumption on the association between MeHg and behavior. The study will be conducted among a birth cohort of 788 infants born between 1993 and 1998 to mothers residing adjacent to a PCB-contaminated harbor and Superfund site in New Bedford, Massachusetts. MeHg was measured in maternal hair collected at approximately 10 days postpartum. Associations with behavioral outcomes will be estimated with multivariable regression, controlling for a wide range of covariates. Results of this study will build upon the literature on the neurodevelopmental effects of MeHg. This is critical to arriving at consensus in informing the public about the potential hazard of prenatal MeHg exposure, given that the primary source of exposure is fish consumption, which is associated with substantial health benefits, including benefits to neurodevelopment. PROJECT HEALTH RELEVANCE The association between prenatal methylmercury (MeHg) and neurodevelopment is not well understood. This study will be an important contribution to the literature necessary for informing the public about the potential hazard associated with prenatal MeHg exposure, given the apparent benefit of fish consumption on the developing brain.
DESCRIPTION: (provided by applicant) Cognitive decline is a common, but not inevitable, accompaniment to aging. While much research is currently being directed at Alzheimer' s Disease -- one of the most severe expressions of cognitive decline -- relatively little work is currently aimed at identifying risk factors and mechanisms associated with early (i.e., subclinical) cognitive decline, even though this may be the stage most amenable to prevention. An important issue that needs to be clarified in relation to the etiology of cognitive decline is the role of environmental lead exposure and the interaction of lead with specific candidate genes. In this application we discuss how four candidate genes -- APOE, the HFE (hemochromatosis) gene, transferrin, and Tau protein -- may interact with lead burden to increase the risk for oxidative cell damage leading to neuronal cell loss and cognitive deficits. We review the last 11 years of our research on low-level lead toxicity and describe compelling preliminary data. We then propose a new study that calls for new data collection in our established Bostonarea cohorts [Normative Aging Study (NAS), Nurses Health Study (NHS), and Community Lead Study (CLS)]. Our specific aims are to test hypotheses related to the impact on cognition of lead burden and its potential interaction with our four polymorphisms of interest. We will look at cognition cross-sectionally and longitudinally, using a validated battery of telephone cognitive function assessment tools as well as a battery of in-person cognitive tests that we have been administering in person since 1993. Results of this research promise to shed further light on lead's potential impact on society and may also give rise to tools for identifying susceptible individuals as well as early brain effects.
Crisp Terms/Key Words: genetic susceptibility, disease /disorder etiology, apolipoprotein E, clinical research, neurotoxicology, gene environment interaction, clearance rate, bone imaging /visualization /scanning, bone metabolism, cognition disorder, neuropsychological test, tau protein, longitudinal human study, toxicant interaction, lead poisoning, transferrin, statistics /biometry, human subject, gene expression, pathologic process, urinalysis, clinical chemistry, hypertension, biomarker, creatinine, human old age (65+)
DESCRIPTION (provided by applicant): Fish and other seafood may contain beneficial nutrients as well as harmful contaminants. The prenatal period is a time of particular susceptibility to the adverse effects of organic mercury as well as to the beneficial influence of nutrients such as elongated n-3 fatty acids and selenium. However, little is known about the balance of risk and benefit from maternal fish intake during pregnancy on child development. The small number of previous studies regarding fish intake and child development are limited by lack of detailed nutrient measures, short duration of follow-up, and few outcome measures. In the proposed study, we will use information on maternal fish intake and biomarkers of mercury and nutrient exposure to examine the combined influence of the risks and benefits of maternal diet during pregnancy on child development. This project build upon the established infrastructure of Project Viva, an ongoing pre-birth US cohort study with stored maternal blood samples, validated prenatal dietary assessment, and detailed information on a number of covariates including home environment and other important predictors of child development. Study aims will be to assess associations of levels of mercury and n-3 fatty acids from maternal blood collected during pregnancy with child cognition and behavior at age 7 years. In addition, we will examine the influence of maternal prenatal fish intake on child cognition and behavior, with and without additional adjustment for mercury and n-3 fatty acids. We will explore the role of nutrients as modifiers of the effect of prenatal mercury exposure, by determining whether mercury has a stronger adverse effect among participants with lower levels of elongated n-3 fatty acids or selenium. Funds from this grant will support wide-ranging cognitive and behavioral assessments added to the already-funded cohort follow-up visit at age 7 years, as well as assays of exposure biomarkers and data analysis. This project is both cost- and time-efficient compared to the initiation of a new cohort, and offers many advantages over previous studies. The study team brings internationally recognized expertise in establishing, maintaining, and analyzing data from a longitudinal cohort, in performing and interpreting assays of nutrient and toxicant exposure, and in interpreting cognitive test results. The proposed study will provide information important to helping mothers and their health advisors make the best decisions about diet during pregnancy to optimize child development. The proposed study will provide timely and high quality data to inform the ongoing national policy discussion regarding the nutritional benefits as well as the contaminant risks from fish intake during pregnancy, and how these competing effects may balance out. Getting the message right is crucial, since pregnant women act upon recommendations regarding the safety of dietary fish intake. The right kind of message could improve children's developmental potential, but the wrong message could do the opposite.
DESCRIPTION (provided by applicant): The developing central nervous system is particularly vulnerable to environmental toxicants such as heavy metals. While lead (Pb) poisoning has been extensively studied, the neurotoxicities and dose-effect relationships associated with other heavy metals, such as arsenic (As) and manganese (Mn), are poorly understood. Data are even more limited on the effects of joint exposures to these metals, even though this exposure scenario more accurately reflects the real world situation. In this project we will build upon an ongoing birth cohort study of 750 mothers and their children at a former hard rock mining community, the Tar Creek, OK, Superfund mega-site. To date, children have been followed to age 2 years, involving the collection of extensive Pb, Mn, and As biomarker data (blood, hair, nails), providing a record of lifetime (including prenatal) exposures to these metals. In the proposed project, assessments will be conducted when children are 5.5 years of age. At that time, Mn and Pb will be measured in blood; Mn, As, and selenium (Se) in hair; and As and Se in nails. The neurodevelopmental assessment will include tests of intelligence, memory, visual- motor/spatial abilities, executive functions, and behavior. Data analyses will model the associations between neurodevelopment and individual metals and between neurodevelopment and joint exposures to metals. Another goal is to determine if genetic susceptibility factors modify the associations between metal exposures and neurodevelopment, focusing on genetic variations in 4 pathways chosen to reflect metal metabolism or synaptic plasticity: iron metabolism (HFE, transferrin, DMT-1), cholesterol metabolism (apolipoprotein E), neurotransmitter metabolism (dopamine transporter, dopamine D4 receptor, norepinephrine transporter, monoamine oxidase, catechol O-methyltransferase, NMDA receptor, 5-HT1A receptor), and methylation metabolism (methylenetetrahydrofolate reductase, GST- M1, T1, P1, omega, purine nucleoside phosphorylase, cytosolic serine hydroxymethyltransferase). This prospective study will provide the most comprehensive data to date on the potential neurodevelopmental correlates of Mn and As exposures, and on the potential interactions among Pb, Mn, and As exposures. This prospective study of children from birth will provide the most comprehensive data available on the potential adverse effects on neurodevelopment of exposure to the heavy metals arsenic and manganese. The effects of exposures to combinations of metals, including lead, is also a major focus. This more closely mirrors "real world" exposures, which invariably involve a mixture of compounds rather than a single compound.
DESCRIPTION (provided by applicant): The developing central nervous system is particularly vulnerable to environmental toxicants. While lead poisoning has been extensively studied and public health measures to reduce exposure implemented, exposure is still high in some populations. Other toxic metals (arsenic and manganese for example) remain elevated in the environment, and the effects of arsenic and manganese on neurodevelopment remain poorly understood. Critical questions also remain regarding individual differences in susceptibility to metals (even lead) and whether combinations of neurotoxicants are synergistically toxic. Given this, the effect of joint exposure to combinations of metals is a critical public health issue, as this exposure scenario is more reflective of the real world situation. The role of iron metabolism genes, which may regulate the metabolism of multiple neurotoxic metals, may also play a key role in understanding the mechanisms by which metal mixtures produce neurotoxicity. In this project we will establish a new birth cohort in Mexico City to measure biomarkers of internal dose for manganese, arsenic and lead and analyze their interactions in predicting associations between metal exposure and neurodevelopment. Furthermore, we will study the modifying effects of iron metabolism gene variants (HFE, Transferrin, Transferrin Receptor, and Divalent Metal Transport Protein-1) on the neurotoxicity of these metals and their influence on placenta! transfer of metals. We will also explore the effect of variants in the GST-M1, T1 and P1 genes on Arsenic neurotoxicity. Our research team is particularly well situated to conduct this work as we have a long-standing collaboration with the National Institute of Public Health in Mexico, enabling us to quickly assemble highly skilled field teams for sample collection and phenotyping. Furthermore, our pilot data suggest that exposure to Mn, As and Pb are elevated in the target population, and that biomarkers of internal dose may be associated with poorer developmental outcomes.
DESCRIPTION (provided by investigator): Children are exposed to a mix of potential neurotoxicants, including metals, via widespread contamination of food, air, soil and, in some settings, water. Exposure to neurotoxicants during early development can be particularly deleterious. Furthermore, the substantial developmental and physiologic changes of adolescence have been shown to be sensitive to early life exposures and experience. Therefore adolescence represents a critical time for manifestations of early life exposure effects but there are few environmental epidemiologic studies of this age group. The primary aims of this study are to assess the relation of early life exposure to neurotoxic metal mixtures, and their interactions, with early adolescent behavior and cognition. The metals to be studied are prenatal manganese (Mn), methylmercury (MeHg), arsenic (As), lead (Pb) and postnatal Pb exposure. We will assess the relation of early life exposure to these metals, individually and jointly, with (1)
behavior and cognition, particularly impulsive or hyperactive behaviors, attention, language skill, executive function, memory and learning skills among 13-year-olds; and (2) prospective changes in behavior and cognition between ages 8 and 13 years. (3) In addition, we will assess the role of prenatal exposure to prevalent organochlorines - polychlorinated biphenyls (PCBs), dichlorodiphenyl dichloroethene (DDE), and hexachlorobenzene (HCB) -- as effect modifiers of metal-behavior/cognitive associations. The proposed study population is a well characterized cohort of 788 children born between 1993-1998, residing adjacent to a PCBand metal-contaminated site and with low-level to moderate metal exposures. Prenatal Pb and organochlorine exposure measures, and archived cord blood, maternal hair and toenail specimens were obtained at birth and the children had developmental assessments from birth through age 8 years. In this study, archived birth specimens will be analyzed for Mn, MeHg, and As and detailed cognitive/behavioral assessments will be performed at age13 years on 500 of the children. The relation of metal exposures with neurocognitive skills will be assessed individually and with interaction terms for each outcome. Then, the relation of individual and multiple exposure measures with potentially correlated outcomes will be evaluated in a single multiple outcomes model. The study's behavioral and cognitive assessments will measures skills directly related to the
risk of childhood learning and behavioral disorders. These increasing common childhood disorders represent a growing source of childhood and, in many cases, later life morbidity and public health concern. The proposed study provides a unique opportunity (given extensive prospective exposure/developmental information and specimen archives already available) to gain insight into potentially remediable risk factors for common behavioral and learning disorders of childhood at a time, adolescence, when there may be increasingly evident functional and behavioral manifestations of early life neurotoxicant exposures.
DESCRIPTION (provided by applicant): We respond to PAR-06-420, jointly issued by the Fogarty Institute and other NIH units, including the National Institute of Child Health and Human Development and the National Institute of Environmental Health Sciences. Basic and clinical researchers of the University of Massachusetts Medical School (UMMS), Auburn University (AU), Harvard University (HU), and the Universidade Federal do Para (UFPA) (Brasil) request funding to develop an interdisciplinary program of translational biobehavioral research and research training focusing on neurotoxic effects of heavy metal exposure in humans and animal models. In this first project, the primary focus will be on effects of Hg exposure (emphasizing initially on methymercury [MeHg]). The project will (a) validate novel methodology for evaluating neurobehavioral functioning in nonhuman primates, (b) apply the methodology to study effects of Hg exposure on behavior, and (c) extend the methodology to evaluate sensory and neurocognitive function of Hg exposure in children living near gold-mining operations in Brasil. The grant will permit a three-pronged program: (1) In the first year, collaborating investigators will merge streams of research at UMMS, HU, AU, and UFPA to the end of validating a battery of nonverbal neurobehavioral tests that is appropriate for use with both nonhuman primates and human children. (2) In both project years, the project team will conduct feasibility research to apply the battery to assess sensory and neurocognitive effects of mercury exposure in (a) laboratory studies of nonhuman primates and (b) field studies with children. (3) The team will evaluate the feasibility of developing a sustained program of pre-doctoral and post-doctoral research training focusing on interdisciplinary analysis of the neurobehavioral effects of heavy metal exposure. The project will take advantage of successful US program models such as the NICHD- supported Mental Retardation Developmental Disabilities Research Centers. PUBLIC HEALTH RELEVANCE: This project concerns the effects of environmental mercury on the development of the brain and behavior. The project will be conducted in gold-mining areas in the north of Brazil, which are heavily polluted with mercury due to processing of gold ore there. Our main focus will be on the children of gold miners. We hope to assess the impact of the pollution on child development to better inform local decision-makers and to take steps toward development of potentially helpful therapies.
DESCRIPTION (provided by applicant): Methylmercury occurs as a contaminant of seafood and freshwater fish worldwide. Tragic pollution episodes have demonstrated that the fetal brain is particularly susceptible to methylmercury toxicity; the adverse effects on the nervous system appear to be irreversible and are much more widespread and serious in children than in adults, especially when exposures have occurred prenatally. This application builds upon the extensive data already collected on a cohort of 1,022 children born in 1986-1987. Detailed information was collected at the time of birth and during the course of follow-up at ages 7 and 14 years. Our previous research efforts have emphasized impacts during childhood and adolescence caused by developmental exposures to methylmercury as a possible paradigm for developmental neurotoxicant effects. The intention is now to complete the follow-up by examining the cohort members as young adults to determine the possible persistence of mercury-associated deficits in neuropsychological and cardiovascular functions, and the impact on educational achievements and overall health. The specific aims of this project are therefore to determine if methylmercury-associated cognitive deficits affect educational achievements, including high- school diploma and college (bachelor) studies; to examine cohort members at age 21 years to determine cardiovascular functions that may be affected by methylmercury exposure; to carry out analysis for PCBs in banked umbilical cord blood from cohort members, and to determine selenium intake from analysis of blood samples; and to assess genotypes for relevant genes that may affect methylmercury toxicokinetics or defense mechanisms against peroxidation processes that may be initiated by methylmercury exposure.
DESCRIPTION (provided by applicant): Toxic effects to the nervous system may become detectable only after latent a period when the deficits become unmasked as a result of age-related degenerative processes. This hypothesis would take a long time to test in prospective studies, and retrospective assessment of prenatal exposure is bound to be seriously imprecise. This epidemiologic conundrum will now be resolved in the Faroe Islands, where prenatal exposure to methylmercury (MeHg) may be estimated, because the MeHg exposure primarily originated from ingestion of whale meat. Local catches of whale pods varied substantially between islands and from year to year, and dissemination of whale meat was difficult until about 1950, when more advanced transportation between the islands and refrigeration became available. A subject's potential exposure to MeHg prenatally can therefore be classified from the detailed whaling data for the pregnancy period at the place of birth. The Faroese have a very high intake of seafood, but their cardiovascular mortality is nonetheless higher than elsewhere in Scandinavia, and Parkinson's disease is about twice as common as expected. We will therefore examine two groups each consisting of about 250 men and women, who were all born around 1935, when variation in whale catches was the greatest. Matched by sex and age, one group's place of birth will be communities with great availability of whale, and the second group's place of birth will be communities without. Each subject will be approximately 70 years at the time of the study and will undergo a series of neurobehavioral and cardiovascular function tests. Postnatal exposure to MeHg and to polychlorinated biphenyls and related contaminants will be assessed by detailed questionnaire and by analysis of appropriate exposure biomarkers. In addition, essential nutrients from seafood, such as n-3 fatty acids and selenium will also be measured in blood samples. The effects of prenatal and/or postnatal neurotoxicant exposures will be determined after confounder control in statistical analyses that will include structural equation models. These results will provide evidence on long-term health implications of developmental and postnatal exposures to these neurotoxicants, which are widely present in freshwater fish and seafood, and the possible compensatory effects of concomitant intakes of essential nutrients. This study will provide unique evidence on adverse health effects of prenatal contaminant exposure as they appear at an advanced age. The results may therefore provide new information that could very likely have an impact on the preventive efforts and standard-setting of the EPA and other regulatory agencies in the U.S and elsewhere.
DESCRIPTION (provided by applicant): The causes of Parkinson's disease (PD) are poorly understood, and so treatment is currently limited to palliative options. Studies of PD have identified both environmental and genetic factors that contribute to the pathophysiology of PD. We will investigate how three PD-related proteins, alpha-synuclein, parkin/KO8E3.7 (the C. elegans homologue of parkin) and DJ-1, affect the toxicity due to inhibition of complex I of the mitochondrial electron transport chain. We will mainly use rotenone as the model toxin for complex I disruption. The three PD-related proteins each have very different putative functions, and how these activities integrate with the pathophysiology of PD is poorly understood. Our preliminary data indicate that expressing alpha-synuclein or deleting KO8E3.7 increases oxidative damage and apoptosis induced by rotenone treatment in C. elegans. In addition, we have identified compounds, D-beta-hydroxybutyrate (a mitochondrial complex II stimulant), tauro-ursodeoxycholic acid (an anti-apoptotic bile acid) and probucol (a potent anti-oxidant), that each partially inhibit rotenone toxicity when applied alone, but in combination prevent rotenone toxicity in C. elegans. In this proposal, we will determine the mechanism of by which alpha- synuclein, parkin/KO8E3.7 and DJ-1 increase mitochondrial damage in both C. elegans and mammalian neurons (primary mouse neuronal cultures and the human neuronal BE-M17 line). We will also determine whether the treatment strategies identified in C. elegans protect mammalian neurons in vitro and in vivo. We hypothesize that mutations in PD-related genes destabilize the mitochondrial electron transport chain leading to an increased tendency to produce free radicals and activate apoptosis. We also hypothesize that combined use of agents that enhance electron transport and inhibit apoptosis (or oxidation) will protect neurons against degenerative processes associated with PD. Aim 1 will determine whether manipulation of PD-related genes inhibit mitochondrial or proteasomal function, and whether the sensitivity to inhibition increases with age. Aim 2 will determine whether manipulation of PD-related genes increase apoptosis during rotenone-induced toxicity. Aim 3 will determine whether combined use of a mitochondrial complex II stimulant plus an apoptotic inhibitor or antioxidant protects neurons against degeneration in transgenic mice carrying the A53T alpha-synuclein transgene..
Parkinson's disease (PD) is a relatively common, debilitating neurodegenerative disorder of uncertain etiology. Recent studies of PD concordance in twins indicate that PD experienced after the age of 50 years (i.e., > 90 % of PD) must be either largely environmental in etiology, or the result of still undiscovered gene-environment interactions. Exposure to metals, particularly lead, has been associated with the development of PD in a few but highly-suggestive studies. This topic has not yet been studied epidemiologically using state-of-the-art biological marker techniques for measuring metals exposure and accumulation. In addition, there is reason to believe that gene-metal interactions may greatly increase the risk of PD. In particular, some evidence suggests that increased exposure to lead interacts with the altered iron metabolism cause PD. Given that iron metabolism is dependent to as large degree on genetic factors, mutations that alter iron metabolism may be associated with PD. In this application, our research team will take advantage of well- described, highly motivated and geographically convenient populations of PD patients and potential controls, state-of-the-art methods for measuring biological markers of metals exposure, polymerase chain reaction (PCR) assays, assessment of pesticide exposure, and a highly- experienced team of investigators to conduct a new case control study. In this study, we will evaluate PD patients > 50 years old as well as controls matched by age, gender, ethnicity, and geographic region. We will test several hypothesis, the main two of which will be that (1) among the cases and controls, higher levels of lead in bone are associated with a higher odds of being a PD cases; and (2) the effect of bone lead will be highest in individuals with at least one copy of the C282Y or H63d hemochromatosis (HFE) gene mutation. We will also explore several subsidiary hypothesis related to the relationship of PD to levels of manganese and copper in toenails and to pesticide exposures, assessed by self-reported history and by linking job histories with a job- exposure matrix.
Crisp Terms/Key Words: human middle age (35-64), human old age (65+), biomarker, polymerase chain reaction, disease /disorder proneness /risk, gene mutation, human subject, questionnaire, metal metabolism, copper, manganese, metal poisoning, lead poisoning, nervous system disorder epidemiology, Parkinson's disease, occupational hazard, pesticide biological effect, longitudinal human study, bone metabolism, skin derivative, work site, gene environment interaction, genetic susceptibility, environmental exposure, patient oriented research
DESCRIPTION (provided by applicant): We propose to study prospectively whether the risk of Parkinson's disease (PD) is predicted by plasma levels of organochlorine pesticides, polychlorinated biphenyls (PCBs), and the antioxidant uric acid. The proposed investigation will take advantage of a unique population comprising over 35,000 Finnish men and women who provided blood samples in 1968-72 as part of a health examination survey, and have subsequently been followed for incidence of PD for over 20 years. Between the baseline survey and 1994, 201 new cases of PD have been confirmed in this population. Plasma levels of organochlorine insecticides, PCBs, and uric acid will be assessed among these cases and controls matched by age and gender. Specific hypotheses to be tested include: 1) That higher plasma levels of para, para-dichlorodiphenyl dichloroethene [p,p-DDE], the major metabolite of DDT, dieldrin, hexachlorobenzene (HCB), beta-hexachlorocyclohexane (HCH), heptachlor, heptachlor epoxide, trans-nonachlor, and oxychlordane are associated with increase risk of PD; 2) that higher plasma levels of PCBs (sum of all congeners) are associated with an increased risk of PD; and 3) that higher plasma levels of the antioxidant uric acid are associated with a decreased risk of PD, and exert a protective effect against the deleterious effects of DDE. The proposed investigation is unique and innovative as there are no previous data relating biomarkers of exposure to environmental contaminants and future risk of developing PD. The blood samples were collected at a time of high exposure to both organochlorine insecticides and PCBs as supported by the high mean plasma level of DDE and PCBs that we found in a pilot study. Finally, the long follow-up period will allow the investigation of long-term effects of exposure.
DESCRIPTION (provided by applicant): A significant relationship exists between iron nutrition and cognition and behavior. Behavioral problems are also observed in children with high manganese and manganese neurotoxicity resulting in a Parkinson-like disorder that is widely recognized in workers employed in mining and manganese ore processing. Our recent work has established that iron deficiency enhances olfactory uptake of manganese and promotes accumulation of this toxic metal in the basal ganglia. Thus, the major underlying hypothesis of this proposal is that absorption of inhaled manganese is up-regulated upon iron-deficiency such that neurological complications of poor iron status are compounded by an increased vulnerability to the toxic effects of manganese exposure. The proposed research will contribute fundamental understanding of physiological risks associated with metal-induced toxicity and, more specifically, the interactions between iron status and manganese neurotoxicity. To accomplish this goal we will: 1) Determine the distribution of intranasally instilled manganese in the brain of control and iron-deficient rats using magnetic resonance imaging; 2) Determine motor coordination and learning/memory capacity of exposed and non-exposed cohorts through use of rotorod and bridge-walking as well Morris water maze tests; and 3) Examine CNS damage due to manganese intoxication in control and iron-deficient rats by determining indices of oxidative stress (3- nitrotyrosine and 8-hydroxy-2'deoxyguanosine levels), identifying neuronal degeneration (Fluoro-Jade staining) and evaluating astrocyte viability using anti-fibrillary acidic protein (GFAP) and microglia status using anti-isolectin B4.
DESCRIPTION (provided by applicant): Polycyclic aromatic hydrocarbons (PAH), derived from the incomplete combustion of fossil fuels and tobacco smoke, elicit their toxic actions on tissues primarily through activation of the aryl hydrocarbon receptor (AHR). The AHR is a member of the Per/ARNT/Sim family of transcription factors that regulates 'target' gene expression in response to physiological or pathological (e.g., environmental toxicants) cues. Virtually nothing is known about the expression or function of this protein in the uterus during pregnancy. Expression of AHR at the maternal-fetal interface, coupled with the fact that AHR-deficient female mice have reduced litter sizes, suggests that this transcription factors serves a biological function during pregnancy. AHR-activating ligands, such as PAH, have been implicated from both epidemiological and animal studies to have deleterious effects on reproductive function. From our own in vivo dosing scheme, we know that PAH profoundly effect pregnancy when administered early in gestation. Gene profiling experiments using uterine decidual tissue of early pregnancy demonstrate that wholesale repression of several cell cycle genes occurs in response to PAH treatment in vivo. A corresponding decrease in decidual stromal cell mitosis was observed. Computer-based sequence analysis identified consensus AHR response elements in the promoters of many of these cell cycle genes, suggesting possible involvement of AHR as part of the gene repression mechanism. Interestingly, AHR protein expression through early gestation mirrors that of two type I histone deacetylases (HDAC) and AHR was shown by immunoprecipitation to form a physical interact with the HDAC2 in vivo. It is hypothesized that selective repression of several cell cycle regulatory genes in the uterus by PAH occurs through epigenetic measures involving an interaction between the AHR and histone deacetylases. If found to be the case, this would suggest that hazardous chemicals found essentially everywhere in our environment can epigenetically alter the expression of genes on a global scale through histone modifications. Furthermore, we believe that AHR functions as an environmental sentinel to signal cell cycle arrest when cells are exposed to environmental toxicants such as PAH.
DESCRIPTION (provided by applicant): Inorganic Arsenic is a toxic compound with significant public health impact. The Environmental Protection Agency (EPA) has identified 1,300 sites on its National Priorities List (NPL), and arsenic has been found in at least 781 of these sites. Arsenic is also a by-product of coal combustion, as well as a naturally occurring water contaminant in many regions of the world, including the USA, Exposure may occur by a variety of pathways including inhalation of dusts in air, ingestion of contaminated soil or water, or through the food chain. Arsenic has been associated with a number of adverse health effects. However, the precise relation of arsenic to pregnancy outcomes has not been established. Thus, we wish to extend the previous work we conducted in Taiwan and Bangladesh to an assessment of birth outcomes in a prospective, repeated measures study of expectant mothers and their newborns in Bangladesh. Currently, an estimated 133 million people in Bangladesh are at risk of disease from drinking arsenic-contaminated drinking water. The proposed studies will evaluate standard birth outcomes at exposure levels that are relevant not only to the U.S. population, but also globally. The proposed studies will assess this risk in a population with a wide range of exposure, from low to high. Together, these data will add substantially to the existing risk assessment information by elucidating birth outcomes after arsenic exposure; the role of methylated forms of arsenic in the urine as biomarkers of exposure and risk; and an evaluation of a new potential marker of adverse outcome (proteomic profiles), as well as the influence of candidate genetic susceptibility traits as risk modifiers. This project is relevant to the overall strategic plan of the NIEHS in several ways. Firstly, we will examine a range of health effects of a significant environmental toxicant, arsenic. Secondly, we will define human biomarkers of exposure, early effects, and genetic susceptibility to arsenic exposure. Thirdly, we will examine exposure-response relationships for arsenic-induced birth outcomes. Fourthly, we will incorporate new, sensitive toxicogenomic technology (proteomics) to assess potentially novel biomarkers of exposure and effect in a molecular epidemiologic setting. Lastly, the study is international, sited in the developing world. The proposed human studies will fill important research gaps in our knowledge of arsenic toxicity and inform clinical and public health interventions.
DESCRIPTION (provided by applicant): The proposed study is a competing continuation of R01 ES09718, 'Environmental Organochlorines and semen quality'. There is scientific and public concern about whether several families of chemicals considered endocrinologically active, such as PCBs and phthalates, have adverse male reproductive health effects. Concern stems from studies showing that a majority of the U.S. general population are exposed to PCBs and phthalates, as well as animal and limited human studies suggesting possible associations of PCBs and phthalates with adverse reproductive health effects. In the proposed competing continuation, we will build upon our earlier work and more precisely define the exposure-response relationships we found between estrogenic and anti-estrogenic PCBs and alterations in semen parameters and reproductive hormones. We will extend our investigation to include measurements for hydroxylated metabolites of PCBs (OH-PCBs) since studies report that OH-PCBs are prevalent in human serum and may be more biologically active than the parent PCBs. The proposed new research direction on phthalates and male reproductive health evolved from a pilot study conducted in collaboration with the CDC. Long-standing toxicological data consistently shows that select phthalatesare male reproductive toxins. Leveraging the ongoing study on organochlorines and semen quality, we analyzed urine samples for phthalate monoesters from a sub-group of men. The results were suggestive of an association between select phthalates and alterations in semen parameters and reproductive hormones, as well as increased sperm DNA damage measured with the comet assay. The apoptosis DNA diffusion assay will be incorporated into the proposed study since the measurement of apoptosis is a necessary counterpart to the assessment of sperm DNA damage, as many cells with significant DNA damage are eliminated by apoptosis. To determine the optimal design for assessing exposure to phthalates, we conducted an exposure study on a sub-sample of men from the organochlorine and semen quality study. Each subject collected nine urine samples over three months. Variance components for between-subject and with-in subject, consisting of monthly and daily variances, were calculated. Based on our results, the optimal exposure assessment strategy for the proposed study is to collect three urine samples from all subjects on day 1, 2 and 31 of their study participation.
DESCRIPTION (provided by applicant): Male reproductive disorders have become an important public-health issue during recent decades: The incidence of testicular cancer has increased and semen quality has decreased, and the incidences of cryptorchidism and hypospadias appear to have increased in certain regions. These conditions are thought to reflect an underlying entity called the testicular dysgenesis syndrome (TDS), with a common origin in fetal life. TDS appears also to be related to other adverse effects, such as decreased birth sex ratios. Although the etiology is uncertain, endocrine disruption is suspected, perhaps due to persistent organochlorine pollutants. Research in this area has been impeded by the relative rarity of some pathologies and because functional assessment of male reproduction is feasible only at adult age, i.e., many years after the initial toxic exposures. We have chosen to carry out a study in the Faroe Islands north of Scotland, where the residents have an increased exposure to suspected endocrine disrupters from traditional marine food, which may include meat and blubber from the pilot whale. Wide exposure intervals are known to exist and appear independent of confounders in this rather homogeneous community. In Faroese adults, an estimate of prenatal exposure levels can be obtained from the mother's dietary habits and current serum concentration of persistent organochlorine substances. The study will therefore examine reproductive outcomes thought to be associated with endocrine disruption and the TDS. These will include measures of semen quality, ratio of X/Y chromosome-bearing sperm, chromatin stability of the sperm, sex ratio of births, and waiting time to pregnancy. The study will include 400 pregnant couples whose mothers are also invited to participate. Because the men of the pregnant couples constitute a selected population (about 250 are expected to provide a semen sample), we will also examine semen quality parameters in 250 young men from the general population (aged 20-24 years) as well as their exposures. This study therefore aims at providing evidence whether increased exposure to endocrine disrupters, whether from postnatal sources or from prenatal exposure caused by maternal seafood consumption, is associated with reproductive abnormalities in both men and women. Decreased sperm quality, decreased fertility, and decreased sex ratio are reproductive outcomes of substantial public health interest. Current evidence already suggests that these pathologies may already affect many populations, and the present proposal aims at identifying whether persistent organochlorine pollutants contribute to these abnormalities.
Crisp Terms/Key Words: endocrine disrupting compound, environmental exposure, marine toxin, generation difference, disease /disorder etiology, clinical research, reproductive system disorder diagnosis, aquatic organism, international cooperation, testis disorder, reproductive development, early experience, embryo /fetus toxicology, pregnancy, longitudinal human study, epidemiology, chlorohydrocarbon insecticide, human pregnant subject, human subject, British Isles, sex chromosome, sperm, semen, fertility, food chain contamination, biomarker
DESCRIPTION (provided by applicant): It is clear that the disease process known as asthma is on the increase in the United States and other developed countries. This increase in the incidence of asthma is not due to changes in the genetic makeup of the population, but rather a combination of environmental factors. Some of the triggers for asthma have been delineated, as well as the actual molecules that drive the inflammatory response. However, the full spectrum of the inflammatory mediators responsible for the initiation and propagation of asthma have yet to be fully defined. We have established a novel model of murine asthma-like pulmonary inflammation based on house dust from the homes of children with asthma. In this application we will build on this novel model to provide closer documentation of the initiators and propagators of the asthma like response. We will pay particular attention to the mechanisms of how these initiators trigger an asthmatic response. Perhaps of greater importance, we will define the inflammatory molecules upregulated by the environmental triggers with particular emphasis on tumor necrosis factor (TNF). The application will focus on the triumvirate of endotoxin, allergens, and outdoor pollutants both singly and in combination. The first specific aim will look at the role of endotoxin in triggering the asthmatic response. This specific aim will use a combination of removing endotoxin from the house dust in addition to examining endotoxin tolerant animals. Using more than one approach increases the probability of success, as well as enhances the rigor of the observations. In the second specific aim, we will remove the allergen from the house dust extract. In the third specific aim we will determine the potential for concentrated air particles, which represent outdoor pollutants, to exacerbate the asthmatic response. Additionally, we will determine whether exposure to the concentrated air particles will prime an animal to develop an asthmatic response. For each of these specific aims we will not only determine if inflammation is present, but also define the range of inflammatory mediators. In our last specific aim we will integrate the data from the previous work to decrease the asthmatic response by blocking TNF by multiple modalities. The results from these studies will define the participation of endotoxin, allergens and outdoor pollutants in the pathogenesis of asthma and determine the role of TNF in causing the inflammation. Lay language: This application will examine how house dust causes asthma. It will specifically examine how 2 different components found in the house dust, cockroaches and bacteria, interact with outdoor air pollution to result in pulmonary inflammation. We will also investigate if a new drug approved for the treatment of rheumatoid arthritis will also help prevent asthma.
DESCRIPTION (provided by applicant): The Problem: Hospital admissions for pneumonia are increased by elevated air particle levels. The mechanism(s) underlying particle effects on lung infection are unknown, but may reflect increased incidence of infection, increased severity of infection, or both.
Hypothesis: The pathogenesis of the pneumococcal pneumonia (the most common variety and the disease we will study) suggests three possible mechanisms for particle effects: enhancement of lung cell 'receptors' used by bacteria for initial adhesion, damage to antimicrobial function of host cells (AMs and PMNs), and exaggerated inflammation in established infection leading to worse signs and symptoms. Hence, the central thesis of this research is that oxidant components of air particles mediate 1) dysfunction of host defenses against infection (incidence) and 2) increased inflammation in extant pneumonia (severity). Experimental Plan: Aim 1 will measure expression and function of pneumococcal 'receptors' (e.g., PAF receptor) used by pneumococcal for initial adhesion after exposure to concentrated ambient particles (CAPs) or control particles. Aim 2 will determine effects of air particles on pulmonary inflammation before and after onset of pneumococcal pneumonia. The hypothesis to be tested is that particles cause enhanced release of cytokine mediators by primed AMs, leading to increased inflammation and ultimately oxidant damage to both AM and PMN In vivo and in vitro studies will measure release of pro-inflammatory cytokines, cell influx and viability and severity of pneumonic inflammation. Aim 3 will test the hypothesis that particle exposure inhibits bacterial clearance via oxidant-dependent damage of anti-microbial functions of AMs and PMNs. Component analysis will be performed using a panel of CAPs samples to provide links of particle constituents (e.g., metals, organics, endotoxin) with biologic effects. Rotated factor analysis will be used to correlate source types with CAPs toxicity. Specific intracellular oxidant pathways will be identified by measurement of oxidant production, intracellular levels of antioxidants, and the effect of a panel of anti-oxidants and other inhibitors. Significance: This research is relevant to the public health question of how inhaled particles cause pulmonary health effects and to the pathophysiology of lung host defense against environmental agents.
DESCRIPTION (provided by applicant): Particulate matter (PM) attributable to traffic and other combustion sources has been associated with cardiac- related health outcomes, respiratory disease, lung cancer, and overall mortality. Experimental evidence in animals and humans suggests that exposure to traffic-related PM results in systemic inflammation, oxidative stress and cell cycle dysregulation, as well as direct genotoxicity. However, the precise mechanisms are poorly defined and have not been well characterized in large "real-world" populations. We propose to prospectively characterize personal exposures to vehicle exhaust while concurrently collecting repeated blood and urine samples in a subpopulation of the Trucking Industry Particle Study, an ongoing national epidemiologic and exposure assessment study. We will assess relationships between measured exposures and biomarkers of exposure and oxidative and inflammatory effects. Specifically we will measure serum levels of 1, 2 naphthoquinone and 1, 4-naphthoquinone albumin adducts and urinary naphthols (biomarkers of traffic- related exposure) and serum levels of C-reactive protein, interleukin-6 and soluble intercellular adhesion molecule-1 and urine levels of 8-hydroxy-2'-deoxyguanosine (biomarkers of effect) with concurrent exposure measurements. Exposure measurements will include elemental carbon, naphthalene, and bound polycyclic aromatic hydrocarbons (PAH), a novel specific indicator of PAHs attached to elemental carbon particles that can concentrate and transport them into the target areas of the lungs and internal organs. Finally, we will perform genome-wide gene expression micro array analysis using whole blood RNA to identify genes and gene networks that are differentially activated in response to vehicle exhaust exposure. We anticipate that the results of this project will provide novel and comprehensive insights into the normal biologic response to vehicle exhaust, the relevant etiologic agents of exposure, and will further our understanding of those processes implicated in the development of lung cancer and heart disease. These issues are of great public health importance due to the large proportion of the general population exposed to vehicle exhaust on a daily basis. Furthermore, our proposed aims are consistent with the goals of NIEHS Environmental Genome Project (EGP) to improve understanding of human genetic susceptibility to environmental exposures PUBLIC HEALTH RELEVANCE: Particulate matter (PM) attributable to traffic and other combustion sources has been associated with cardiac- related health outcomes, respiratory disease, lung cancer, and overall mortality. We anticipate that the results of this project will provide novel and comprehensive insights into the normal biologic response to vehicle exhaust, the relevant etiologic agents of exposure, and will further our understanding of those processes implicated in the development of lung cancer and heart disease. These issues are of great public health importance due to the large proportion of the general population exposed to vehicle exhaust on a daily basis.
DESCRIPTION (provided by applicant): Problem: The health effects of inhaled environmental exposures are determined in large part by the 'first responded cell of the lung, the alveolar macrophage (AM). The important initial contact of AMs with inhaled agents is through members of the 'scavenger1 receptor class A family (SRA), especially MARCO and SRAI/II. The role of these pattern-recognition receptors in lung defense against inhaled oxidants and how they function for clearance and signaling are important but unanswered questions. Pilot Data: 2 novel and surprising observations spark our proposed research. First, despite being considered as passive 'molecular flypaper1, our data show distinct signaling and modulatory effects of the SRAs on macrophage behavior. Second, SRA-deficient mice show markedly increased inflammation in response to both an inhaled oxidant gas (ozone) and the soluble fraction of particulate air pollution (in addition to known SRA interactions with solid particles). We draw upon our pilot studies and the available literature to formulate our central hypothesis: AM scavenger receptors protect the lung directly by clearance of pathogenic particles and indirectly by removal of the pro-inflammatory oxidized lipids generated within alveolar lining fluid. Specific Aims: Aim 1: This aim will test the hypothesis that SRA deficient mice will show increased lung inflammation and injury in response to oxidant challenges in all 3 physical forms: solid particles (e.g. concentrated ambient air particles, CAPs), liquid (the soluble fraction of CAPs), and gaseous (e.g. ozone). Aim 2: This aim will test the hypothesis that AMs from SRA-deficient mice will show diminished uptake of oxidized lipids generated in lung lining fluid and altered responses (cytokine release, toxicity) compared to wild-type AMs in vitro. Co-culture experiments with lung epithelial cells will test the prediction that diminished clearance of oxidized lipids by SRA-deficient AMs will reveal inflammatory and toxic responses in epithelium. Aim 3: This aim will characterize signaling mechanisms for SRA-mediated particle phagocytosis and AM modulation, including identification of co-receptors and signals for particle internalization, and expression profiling of genetic programs triggered by SRA ligation. Significance: This research will identify novel mechanisms for lung defense against inhaled oxidants and illuminate basic mechanisms for phagocytic clearance of inhaled particles.
DESCRIPTION (provided by applicant): Obesity is an important public health problem that is a risk factor for cardiovascular disease, type II diabetes, some forms of cancer, and for asthma. Preliminary data indicates that the obese are also "at risk" in terms of their susceptibility to air pollution, one of the triggers for asthma. The purpose of this proposal is to use animal models of obesity to examine the mechanistic basis for the relationship between obesity and asthma, using pulmonary responses to the air pollutant ozone (O3) as the outcome indicator. Preliminary data indicates obese mice have greater O3-induced airway inflammation and airway hyperresponsiveness than lean mice. Our hypothesis is that the increased systemic inflammation of obesity, particularly elevations in IL-6 and TNFa, prime lung cells to respond to inhaled pollutants with greater inflammatory responses and enhanced changes in lung function. Moreover, we hypothesize that this systemic inflammation is adipose tissue derived. Two types of obese mice will be employed, both on a C57BL/6 background: Cpefat mice and mice on high fat diets. Lean and obese mice will be exposed to filtered air or O3. After exposure, pulmonary mechanics and airway responsiveness to methacholine will be assessed, bronchoalveolar lavage (BAL) performed, and BAL markers of injury and inflammation measured. RNA will be prepared from the lungs and abdominal fat and analyzed for inflammatory gene mRNA expression by real time PCR. Serum will also be analyzed for markers of obese systemic inflammation. In aim 1, we will determine whether obesity-related increases in responses to O3 correspond temporally with the expression of adipose tissue inflammatory genes, particularly IL-6 and TNFa, during the development of obesity. In aim 2, we will ablate IL-6 and TNFa genetically or with antibodies, and increase systemic IL-6 and TNFa experimentally, to examine the role of IL-6 and TNFa in the effects of obesity on lung responses to O3. In aim 3, we will examine the hypothesis that macrophages that infiltrate adipose tissue of obese mice are the source of the inflammatory molecules that augment airway responses to O3 in obesity. To do so, we will lethally irradiate mice to eradicate hematopoietic stem cells and then reconstitute them with fetal liver cells from genetically altered mice. Understanding the mechanistic basis for the augmented pulmonary responses to air pollution may lead to therapeutic strategies for reductions in responses to air pollution in this at risk population.
DESCRIPTION (provided by applicant): The structural and functional integrity of the lung extracellular matrix (ECM) is largely dependent on the conversion of soluble collagen and elastin to insoluble, fibrous aggregates catalyzed by lysyl oxidase (LO), a copper [Cu(II)] dependent enzyme. This catalyst oxidizes lysine residues within these proteins to generate covalent cross-linkages stabilizing the ECM. Thus, LO plays a central role in the lung morphogenesis and tissue repair. Cadmium (Cd) is a toxic metal for humans. Inhalation and accumulation by the lung of Cd either from environmental contamination or from cigarette smoke induces perturbations of the metal ion homeostasis, which may be a key mechanism for the pathogenesis of the lung. Preliminary studies showed that development of Cd resistance (CdR) of rat lung fibroblasts (RFL6) following long-term Cd exposure was accompanied by upregulation of cellular metallothionein (MT) and glutathione (GSH), and downregulation of LO and its collagen and elastin substrates. These findings led to a hypothesis for the mechanisms of Cd injury to the lung ECM relevant to emphysema pathogenesis: During long term exposure to Cd, the lung fibroblasts upregulate the synthesis of MT and GSH. These intracellular thiols bind Cu ions with a higher affinity than Cd thus severely perturbing the homeostasis of Cu, limiting its availability to LO, and contributing to the downregulation of LO at the mRNA, protein and catalytic levels as shown in preliminary studies. Downregulation of LO would in turn inhibit the crosslinking of collagen and elastin, favoring their destabilization, solubilization and eventual degradation and interfering with their repair. The resulting solubilized collagen and elastin would inhibit their own synthesis possibly by a feedback mechanism, further disturbing the balance between synthesis and degradation of these proteins and disrupting the ECM, events which are characteristic of the development of emphysema. The following Specific Aims are designed to test this hypothesis: 1) To assess mechanisms of Cd perturbation of Cu(ll) homeostasis in CdR-RFL6 cells; 2) To investigate mechanisms of LO downregulation at transcriptional, translational and posttranslational levels in CdR-RFL6 cells; 3) To explore LO effects on the downregulation of its collagen and elastin substrates and on the elastin repair in CdR-RFL6 cells; and 4) To demonstrate elevation of cellular MT and GSH, perturbation of Cu homeostasis and downregulation of LO as key mechanisms in emphysema pathogenesis of rats receiving Cd by chronic administration. The outcome of the proposed research is expected to define key aspects of Cd modulation of LO gene expression and processing by perturbation of Cu homeostasis in the lung, thus enhancing our understanding of the molecular mechanisms for Cd emphysema pathogenesis.
DESCRIPTION (provided by applicant): Environmental or intentional exposure to a broad variety of chemical agents can alter normal endocrine function. The effects of these "endocrine disruptors" (ED) can have serious health implications including deleterious effects to reproductive capacity, fetal development, the immune system, and carcinogenesis. Current animal tests are expensive, use a large number of animals, and are not necessarily applicable to humans. Thus, a validated, human in vitro method to identify ED is an area of great importance. This research project will develop such a test method. Phase I research will prevalidate MatTek's organotypic vaginal- ectocervical (VEC-FT) tissue model for use in identifying endocrine disruptors (ED). A battery of 25 model compounds with known ED activity will be used. Changes to key hormonal receptors, enzyme activity, tissue structure, proliferation, and gene expression will be monitored. These data will be used to develop a prediction model that will be a sensitive and specific predictor of ED potential. During Phase II, the test method will undergo formal validation in a multi-center, GLP study. The prevalidated in vitro test to be developed for assessing the endocrine disrupting potential of chemicals/ formulations will have enormous environmental and public health significance. Evaluation of endocrine disruptors is important to minimize hazards to humans and wildlife exposed to chemicals that interfere with normal hormonal regulation. The proposed human organotypic tissue based assay will provide a sensitive and specific assay method to screen a large number of endocrine disrupting chemicals at a reduced cost.
DESCRIPTION (provided by applicant): Radiation Monitoring Devices, Inc. (RMD) proposes to develop a field-portable instrument for the detection and identification of pathogenic viruses that are collected by an aerosol concentrator. It will incorporate modular, microfluidic platforms designed to capture and concentrate the sample for a rapid, two-stage analysis. The first employs pathogen capture on antibody-decorated, paramagnetic nanospheres and will provide rapid, preliminary warning based on the hydrodynamic properties of the nanoparticles when the targeted viruses are bound. This stage will monitor the relaxation of the magneto-optical birefringence of the nanoparticles and will trigger an alarm when the birefringence relaxation rate drops off dramatically. The methodology allows for continuous surveillance over virtually unlimited periods. The second stage will provide either confirmation or correction to the preliminary identification. It is based on the ability to interrogate the captured nucleic acid with a series of unique probes that change their fluorescence properties when they hybridize with specific nucleotide sequences of suspected viral genes. We will compare the speed and sensitivity of detecting these genes by fluorescence generated from molecular beacons, and by the temporal correlation of fluorescence issued from tagged probes. The detection method exhibiting the best performance will be incorporated into the sensor. In each case, Nucleic Acid Sequence Based Amplification (NASBA), an isothermal nucleotide amplification scheme, will be employed to raise the number of target template copies to detectable levels. The biosensor will integrate the two stages of sample transfer, amplification, and analysis in a microfluidic architecture that enables high throughput processing and parallel detection of multiple probes. Employing both proportional mode avalanche photodiode (APD) detectors and Geiger-mode muAPDs that are responsive to extremely low light levels, sensitivity to low virus titers will be possible.
This proposal solicits funding for developing an instrument to detect and identify pathogenic viruses in a rapid, automated two-step analysis that can be used by emergency workers who are responding to a suspected release of airborne (waterborne) agents. The first step screens the sample to determine if there are any virus particles that possess a targeted group of exposed coat proteins. If the answer is affirmative, the virus is isolated and its nucleic acid is analyzed to determine if it carries pathogen genes.
DESCRIPTION (provided by applicant): US Department of Transportation and international regulations require testing of all chemicals for skin corrosivity, which is defined as destruction of human skin at the site of contact. For safe handling and transportation purposes, each corrosive is assigned a "packing group" designation, which depends on how quickly skin damage occurs. Previously accepted test methods have utilized rabbits to determine packing groups. However, corrosive chemicals are very harsh and can cause considerable discomfort and/or severe pain to the animals. A number of in vitro tests have been developed but none of them adequately meet all of the US regulatory and market needs. The goal of this project will be to utilize a human skin-like in vitro model, EpiDermTM, to develop and validate an in vitro assay to determine skin corrosivity packing groups. Phase I research will determine the assay parameters necessary to differentiate between the 4 corrosivity packing groups. A preliminary prediction model (PM) will be developed using 12 reference chemicals and PM will be tested with an additional 28 test materials. After fine tuning the PM, interlaboratory reproducibility and interlaboratory transferability of the assay will be assessed. Successful completion of the Phase I goals will constitute the pre-validation process for proceeding to formal assay validation studies in Phase II. PUBLIC HEALTH RELEVANCE: Skin corrosivity testing is necessary to insure the safe handling and transport of chemicals. Animal-based test methods suffer from animal welfare concerns and current in vitro methods do not meet all US regulatory requirements. This project will develop an in vitro assay to determine skin corrosivity packing groups and fulfill all testing and regulatory needs.
DESCRIPTION (provided by applicant)
Numerous pollutant compounds are continuously released into the environment during their production, use and disposal. The resulting mixtures contaminate the environment and can be a potential threat to human beings. Heavy metals are significant environmental pollutants because they tend to persist in the environment, tend to bioaccumulate and can result in adverse health effects when ingested or inhaled. These metal species can be measured in human tissue or fluids such as blood or urine, usually in central laboratories using complex methods such as high-performance liquid chromatography inductively coupled plasma mass spectroscopy. However, in view of the high labor and analytical costs and long time delays associated with centralized laboratory analyses, there is an immediate need for a portable and inexpensive system for on-site monitoring of exposure to heavy metals in living systems, especially humans, as a result of exposure through drinking water, environmental, and industrial sources. Field deployable monitoring systems for in-situ, on-site, personal human exposure assessment will allow realtime monitoring of exposure to trace toxic metals, and facilitate the use of bio-monitoring data by allowing the instrument to be taken to the sample rather than the traditional way of bringing the sample to the laboratory.
The overall objective of this project is to develop a portable, self-contained, easy-to-use monitor for simultaneous on-site measurement of arsenic, cadmium, mercury and lead metals from one urine sample, in near real time, to assess personal exposure. The proposed sensor will combine the high sensitivity of a specialized electrochemical measurement technique with a unique microarray electrode material and configuration. During Phase I, we will investigate sensor design configurations, electrode material composition, and the preliminary operating conditions which will demonstrate the feasibility of the unique microchip sensor prototype for detection of arsenic and cadmium species. At the conclusion of the Phase I/II program, a compact, packaged prototype sensor instrument will have been developed, tested and utilized in pilot epidemiological field trials.
DESCRIPTION (provided by applicant): Among the various functional imaging techniques, SPECT maintains an important role in the study of disease models in small animals as well as in patient care. Many SPECT-labeled imaging probes that have highly specific distributions with very little background are being used to measure a wide range of biological parameters of importance in small animals including substrate metabolism, blood flow, hypoxia, protein synthesis and receptor characteristics. The utility of this important modality has been significantly enhanced in recent years by the development of methods to image transgene expression in vivo. Furthermore, SPECT is capable of dual-isotope imaging for correlating two biological processes with a single imaging study. Recent demand for small animal SPECT has also been driven by the pharmaceutical industry where in vivo quantification of biological processes to measure an agent's mechanism of action and its concentration at the site of action is necessary. Each of these applications requires excellent spatial resolution, not only because of the small scale of the details to be imaged but also for the demanding detection and estimation tasks. Unfortunately, existing SPECT instrumentation does not provide the required performance, so the development of a high-resolution detector is essential for the improvement of future SPECT systems. To address these issues, we propose to develop a novel detector based on a unique design of a scintillator coupled to a very high spatial resolution readout. Specifically, the scintillator design and fabrication will minimize the loss of resolution arising from parallax errors due to depth-of-interaction effects within the scintillator, while maintaining very high efficiency for the detection of incident radiation. This combination of scintillator and readout, when coupled to a custom designed collimator, will form a SPECT detector module that can achieve extremely fine spatial resolution and high sensitivity in a cost-effective manner. The effort subsequent to the successful development of the detector module will focus on the development of a complete small-animal SPECT system as a prototype product.
DESCRIPTION (provided by applicant): Environmental toxicants are increasingly linked to adverse birth outcomes such as low birth weight, preterm birth, birth defects and developmental delays. Inner-city populations are at higher risk for adverse birth outcomes and also are more likely to be exposed to environmental toxicants. Obstetric providers are seeking clarification about the true risks of many environmental exposures and what message they should deliver to their patients. Patients in this population are confused by the conflicting messages they receive from providers and peers about behaviors and exposures that could put their fetus at risk. The Institute of Medicine and the National Environmental Education and Training Foundation recommend the integration of environmental health concepts into all levels of medical and nursing education. In the proposed SBIR Phase II project called Managing Environmental Risks in Pregnancy, Vida Health Communications, Inc. will complete the development and evaluation of the web-based provider training and patient education multi-media prototyped in Phase I. The final products of this research will be (1) a cross disciplinary web-based training offering continuing education for obstetric providers serving urban populations, (2) an educational DVD in English and Spanish and group discussion guide for showing to patients in clinic and office waiting areas or in facilitated group discussions, (3) a library of colorful support materials in English and Spanish at appropriate levels of literacy for providers to print and distribute to patients made available both as electronic documents (pdf) and in preprinted form. The interventions will be evaluated using focus groups representative of the target audiences. Evaluators will use well-documented qualitative techniques to analyze focus group data. This project will advance the environmental health training of obstetric providers serving urban populations that may be at risk for exposure to substances that can harm their developing fetus. It will also provide much needed education, at appropriate literacy levels, for at-risk women about ways that they can moderate fetal exposure to environmental risks in pregnancy. Narrative: We are beginning to recognize the scope of environmental toxicants that urban-living pregnant women may be exposed to. Given the rapid development of a fetus, especially in the early months of pregnancy, avoidance of exposure is at present the most effective way to mitigate potential harm. This research will contribute to knowledge of how prenatal care providers can interest, inform and motivate pregnant women who likely bear the greatest risk of exposure, so they can protect the fetus and improve the chance of giving birth to a healthy, normal infant.
DESCRIPTION (provided by applicant): Cytochrome P450 (CYP) enzymes catalyze the majority of known drug metabolism and the bulk of these reactions occur in the liver. As a consequence, many clinically relevant drug-drug interactions are associated with inhibition and/or induction of a specific CYP enzyme. Modifications of CYP activities can have profound effects on therapeutic efficacy and can lead to life-threatening toxicity. In order to provide an early warning system for potential serious side effects, detection of specific CYPs responsible for drug metabolism and drug-drug interactions is a goal of all pre-clinical studies. An ideal CYP assay should be rapid, robust, reliable, reproducible, and amenable to automation in multiwell plate formats. This research will develop a microplate format high throughput whole zebrafish CYP functional assay for assessing drug metabolism and drug safety. Based on genetic and physiological similarity to humans, zebrafish show promise as an efficient, predictive animal model for assessing drug metabolism and drug safety. PUBLIC HEALTH RELEVANCE: Project Narrative The zebrafish assay will facilitate detection of specific cytochromes responsible for drug metabolism and drug-drug interactions, and provide an early warning system for potential serious side effects,
DESCRIPTION (provided by applicant)
An inexpensive disposable sensing element that is worn as a vapor exposure monitor or used for urinalysis is proposed for real-time monitoring of pesticide exposure. This sensor, coupled to a battery operated reader, is a direct need for the NIH Exposure Biology Program and has the potential for detect other exposures as well. The sensor is based on Surface-Enhanced Raman Spectroscopy (SERS), which allows precise detection of chemicals that adsorb strongly to roughened SERS sensors, like pesticides, into the high ppt range. The SERS sensors, vapor deposited gold films with subsequent electrochemical roughening, are inexpensive to make, operate in vapor phase under a wide humidity range and in solution. As the sensors are tuned to the analytes of interest, interferences from more concentrated chemicals is limited. In this program, the SERS sensors would be worn as badges to monitor and quantify daily pesticide exposure to a worker by irreversibly binding any pesticide that comes in contact with the SERS sensor. The sensors would also measure pesticide metabolites in urine, providing the NIH with more data on the daily exposure/daily ingestion of pesticides. The SERS sensors will be read using a battery operated Raman instrument with keyed sampling chamber that ensures the SERS sensing elements are automatically aligned into the system. The compact system would be fully automated and incorporate a spectral library with chemometric analysis. Identification and quantification of any detected pesticides would be reported to the field technician. A single reader can be used for evaluation of hundreds of participants.
The core concepts of this technology have been developed and demonstrated. Literature results show SERS detection of organochlorine pesticides and EIC Laboratories has detected organophosphate pesticides in vapor and aqueous phases. Field detection of trace levels of explosives has been demonstrated by EIC Laboratories. The two year Phase I program is designed to expand the pesticides studied, demonstrate detection to CDC listed limits, show an adequate usable field lifetime for the SERS sensing elements, design a fieldable reader, perform a detailed interferences analysis and demonstrate that variances in the urine matrix will not affect the SERS measurement precision. Commercial: EIC Laboratories will commercialize the technology by manufacturing the SERS sensors and battery operated Raman readers. The components will be sold commercially through In Photonics, an affiliated company under common ownership.
DESCRIPTION (provided by applicant): The use of chlorine for the disinfection of domestic water supplies produces a range of halogenated organic compounds as a result of chemical reactions between the chlorine and naturally occurring organic material (NOM). These disinfection byproducts (DBFs) occur at parts per billion level and the consumption of chlorinated water containing these compounds has been linked to an increased incidence of a number of cancers and other ailments. As a consequence, it is highly desirable to reduce the levels of these DBFs in drinking water in order to limit their impact on human health. Current methods for DBP removal tend to focus on the reduction of NOM prior to the chlorination process. However, available technologies are only moderately effective or are cost prohibitive for large-scale applications (e.g. reverse osmosis). Triton proposes to use selective adsorbents to remove the DBFs after the chlorination step. During the Phase I project, it was demonstrated that certain hydrophobic nanoporous sorbents have great advantages over existing materials for a cost effective and efficient removal of DBFs. This Phase II proposal addresses the experimentation for both POU and POE devices necessary to qualify the adsorbent material, relative to standards set forth by both the EPA and the National Sanitation Foundation (NSF). Importantly, this proposed effort also addresses the commercialization of these materials in POU and POE devices with existing companies in the drinking water treatment industry.
The proposed Triton's approach of adsorbing DBFs with a special adsorbing bed of material in both POU and POE devices has the following advantages:
Municipal treatment facilities can continue to use time-tested halogens as the primary and/or secondary means of disinfection, saving billions of dollars in infrastructure required to convert to new water treatment methods such as UV and Ozone. The retrofitable nature of the adsorbent media into POE and POE devices permits suppliers of water treatment devices to expand their market offering without the need to build new devices. Consumers can readily have access to the new technology through the existing marketplace infrastructure and devices.
Crisp Terms/Key Words: cost effectiveness, nanotechnology, evaluation /testing, technology /technique development, water treatment, water pollution, adsorption, waste treatment, hydropathy
DESCRIPTION (provided by applicant): The goal of this proposal is to develop statistical methods for environmental health data when the health effects of interest are complex. The Specific Aims are motivated by problems arising in toxicological and environmental epidemiological studies of the health effects of airborne particulate matter. Specific aims of the project are the development of (i) Wavelet-based historical functional data models for assessing high-dimensional associations between exposure and health; (ii) Hierarchical hidden Markov models for analyzing multivariate functional data arising from animal particulate matter concentrator studies; (iii) Methods to address exposure measurement error arising from spatial and temporal misalignment in particulate matter epidemiology studies. We will apply the proposed methods to several data sets for which existing analysis methods do not make full use of the data, including (i) semi-continuous heart- rate variability data from matter in sensitive subpopulations. In the motivating applications, the methods will provide insight into two scientifically pressing issues in environmental health research: the identification of biologic mechanisms of morbidity and mortality of air particles, and identification of pollution sources responsible for observed health effects. More generally, the proposed methods represent advancements in the areas of functional data analysis, hidden Markov modeling, and measurement error modeling that are applicable in a variety of biomedical research settings involving high-dimensional data.
DESCRIPTION (provided by applicant): The broad long-term goal of this project is to understand mechanisms that restrict DNA replication to once-per- cell cycle. Our preliminary studies have identified checkpoint signaling pathways that prevent DNA re-replication. We have termed these pathways 're-replication checkpoints'. Re-replication checkpoints ensure that only one round of DNA synthesis occurs during S-phase and represent a fundamental mechanism for maintaining genomic stability. The 'Specific Aims' of this project are: (1) To test the hypothesis that the Rad9-Radl-Husl (9-1-1) checkpoint protein complex prevents DNA re-replication. (2) To test the hypothesis that perturbation of re-replication checkpoints promotes gene amplification. We will study the effect of 9-1-1 perturbation on re-replication using several complementary approaches. These include: over-expressing the 9-1-1 antagonist, Husl B, RNA interference experiments targeting Rad9 using a novel adenovirus vector we constructed for gene silencing, and studying cell lines from transgenic Husl-null animals. The putative mechanism by which 9-1-1 prevents re-replication will be determined using biochemical approaches including studying 9-1-1-mediated signaling pathways and putative interactions between 9-1-1 and DNA replication factors. Acquisition of methotrexate resistance will be used as an assay to study amplification of a defined genomic locus (the DHFR gene). We will test the effects of perturbing the re-replication checkpoint on DHFR amplification. Microarray-based Comparative Genomic Hybridization (CGH) will be used to identify re-replicating genomic loci in 9-1-1-deficient cells. Many human oncogenes are over-expressed in cancer cells as a result of gene amplification. Therefore, gene amplification is a hallmark of cancer cells and contributes to tumor progression. Additionally, genes amplified in tumor cells can confer resistance to chemotherapies. Therefore, an understanding of factors that pre-dispose to gene amplification will be of prognostic and therapeutic value in the clinic. If gene amplification in tumors results from loss of re-replication checkpoints, these checkpoints could be exploited using drugs or gene therapies. The results of our studies may provide a novel paradigm for mechanisms of gene amplification and will be directly relevant to tumorigenesis in humans.
DESCRIPTION (provided by applicant)
The metal response element-binding transcription factor-1 (MTF-1) is a ubiquitously expressed transcription factor that plays a primary role in the regulation of a variety of genes involved in metal homeostasis, embryonic development, tumor progression, and oxidative stress or hypoxia signaling. The overall objective of this research is to characterize the overlapping role of MTF-1 with Nuclear erythroid 2 p45-related factor 2 (NRF2), the primary transcription factor for regulating the oxidative stress response, in development and transcriptional co-regulation of stress responsive genes. The investigator's goal for this award is to obtain advanced training in molecular and transgenic techniques utilizing the zebrafish as a model organism for developmental toxicology. These techniques will include morpholino knockdown of gene expression, development of transgenic zebrafish lines, gene expression profiling with oligonucleotide microarrays, and advanced bioinformatic techniques for microarray analysis and computational promoter analysis. The investigator will expand background in environmental/ecological toxicogenomics by applying these techniques to the study of transcription factors that play fundamental roles in the cellular response to metal or oxidative stress. This experience will assist the investigator in the future goal of establishing an independent research laboratory, where molecular and genomic techniques will be utilized to continue research in developmental and molecular toxicology and gene regulatory networks. Training during the mentor phase of this award will be directed by Dr. Mark Hahn (Woods Hole Oceanographic Institution, WHOI), an internationally recognized molecular toxicologist with two decades of research experience in the characterization of transcriptipon factors in a variety of model organisms. WHOI is a world-renowned center of scientific research. Under the guidance of Dr. Hahn, and with consultation from Dr. Elwood Linney (Duke University), the investigator will fully characterize the transactivation domain of MTF-1 and generate a germline transgenic zebrafish that inhibits endogenous MTF-1 signaling through the expression of a dominant-negative construct. In addition to learning advanced zebrafish techniques, time in the mentored phase will involve additional training through formal coursework and seminars, as well as further guidance in grantsmanship that will assist the investigator in transition to an independent researcher.
DESCRIPTION (provided by applicant)
Dr. Gregory Wellenius's long-term career goal is to be become an independent investigator capable of leading patient-oriented research efforts in environmental cardiology. During the mentored (K99) phase of this career development award, the candidate will gain expertise in conducting patient-oriented mechanistic research in the setting of the General Clinical Research Center, assessment of vascular endothelial function, statistical methodology, and exposure assessment. The candidate will also establish his own laboratory, collect preliminary data, and refine power calculations during this phase.
Dr. Murray Mittleman, the candidate's current post-doctoral mentor, will serve as primary mentor and oversee all aspects of the candidate's training. In addition, a team of three co-mentors with complimentary areas of expertise will supplement Dr. Wellenius's training in specific areas. Together, the mentor and co-mentors are committed to assisting the candidate reach his research training and career development goals and to ensuring the candidate's successful transition from postdoctoral fellow to independent researcher.
During the independent (R00) phase of this award, the candidate will conduct a prospective, repeated-measures study to evaluate the mechanisms by which particulate air pollution affects autonomic function, inflammation, and vascular endothelial function in a cohort of patients with congestive heart failure. These putative mechanisms have been evaluated in a number of normal and susceptible populations, but never specifically in subjects with congestive heart failure.
Relevance: Hospitalization rates among patients with congestive heart failure are extremely high. Epidemiologic studies suggest that short-term increases in ambient air pollution are associated with increased risk of hospitalization in this population, but the mechanisms of this effect have not been investigated. Understanding the underlying mechanisms may lead to new preventative strategies, new treatments, or improved air quality.
DESCRIPTION (provided by applicant)
This proposal will allow the Principal Investigator (PI) to gain knowledge and skills in Environmental Epidemiology under the direct supervision of a highly qualified sponsor, which will build on and extend his previous expertise in neurobiology, further enhancing his potential to develop into an independent investigator. The PI is trained in cellular and molecular aspects of neurobiology, and has had some exposure to epidemiology, including some classes at the Harvard School of Public Health taken on a part-time basis. During the first 3 years of this proposal, course work is proposed to attain needed skills in statistical methods for environmental epidemiology, molecular and genetic epidemiology, and neuroepidemiology. This course work, combined with the research to be conducted during this proposal will enable the principal investigator to obtain a doctorate in environmental epidemiology from the Harvard School of Public Health, thus enabling him to be a very competitive candidate for a tenure-track faculty position, which is the long-term goal of the candidate.
The environment at the Harvard School of Public Health is an excellent one to accomplish the candidate's goals. The mentor is a recognized leader in the field of environmental epidemiology who has mentored other K-awardees. The school has a vibrant community of epidemiologists of all types, including genetic and neuro-epidemiologists with whom the candidate can interact. On-going seminar series in, among other topics, statistical methods and chronic (including neurologic) disease, provide fertile ground for learning, intellectual interactions, and development of ideas.
The overall scientific goal of this study is to explore the use of novel biomarkers of neurotoxicity. In particular, the candidate proposes to build on his past neurobiology research to examine the effects of lead exposure on psychophysiologic measures of learning in the context of fear and anxiety, as well as the effects of lead on fine motor control as manifested through a new device for sophisticated analyses of handwriting and on autonomic nervous system control of cardiac function. This proposal focuses on lead not only because of the important public health consequences of lead exposure, but also because the extensive research experience with aspects of low-level lead exposure provides a solid foundation from which to explore these relatively new biomarkers of neurotoxicity.
Crisp Terms/Key Words: environmental exposure, behavioral /social science research tag, neurotoxicology, psychomotor function, association learning, fear, psychophysiology, startle reaction, nervous system disorder epidemiology, lead poisoning, hereditary hemochromatosis, human subject, heart function, electrostimulus, auditory reflex, auditory stimulus, biomarker, human middle age (35-64), clinical research, anxiety, autonomic nervous system, electrocardiography, human old age (65+)
DESCRIPTION (provided by applicant)
The purpose of this Mentored Research Scientist Development Award is to support the interdisciplinary career development of the candidate, Dr. Adrienne Ettinger, who is trained as an environmental epidemiologist. This award will support the candidate for a period of mentored research and additional training in scientific and laboratory methods relevant to genetic and molecular epidemiology. This will enable her to subsequently develop an independent, interdisciplinary research program with a focus on transgenerational effects of environmental exposures, including the identification of molecular markers for noncancer (reproductive) health outcomes and analysis of complex gene-environment-nutrient interactions. The proposal outlines a Career Development Plan which includes working with established experts in genetic and molecular biology, endocrinology, toxicology, and advanced statistics, and is enhanced not only by the intellectual and laboratory resources described herein, but also by carefully chosen training activities and a career development involving laboratory experience, didactic instruction, seminars, and both formal and informal exchanges with mentors. The research component is aimed at evaluating if specific genes (VDR, CaSR, PTHR1), involved in the regulation of calcium homeostasis, modify the relationships between maternal lead biomarkers and physiologic parameters during pregnancy and, ultimately, influence fetal and early infant exposures and birth outcomes. An exploratory aim will investigate genomic imprinting and gene expression in the maternal-fetal unit using evolutionary theories of "gestational conflict" to develop a conceptual framework for understanding these complex interactions at the molecular level. The proposed research takes advantage of archived data and biologic specimens from three sequentially-enrolled longitudinal birth cohorts, and a newly-funded forth birth cohort, which provides a unique opportunity to address the scientific aims and makes efficient use of existing resources from an established, ongoing, and productive collaboration between researchers in the U.S. and Mexico and addresses research areas pertinent to the NIEHS areas of special interest: "Single Nucleotide Polymorphisms and Disease Susceptibility" and "Epigenetics: Role in Environmentally Induced Diseases/Dysfunction."
Relevance: Epidemiologic and experimental evidence suggest that environmental agents are reproductive toxicants, but relatively little is known about the biological mechanisms of effect. The objective of this work is to understand how common genetic variants, involved in calcium regulation, may impact toxicant-induced pregnancy and developmental outcomes.
DESCRIPTION (provided by applicant)
The candidate is an epidemiologist and biostatistician with 10 years experience in conducting population-based observational studies. The application describes a 3-year career development plan that will provide the candidate with the formal training and laboratory research experience necessary for her development as an independent investigator in epigenetic epidemiology of chemical exposures in utero and chronic diseases. The candidate's long-term career objective is to develop an academic program of research that combines epidemiologic, molecular, and epigenetic principles to study the complex associations of genes, their expression, and environmental factors in the etiology of cancer. The training component includes mentorship by an environmental cancer biologist and six-month rotations annually in an environmental genetics and epigenetics laboratory in additional to formal course work education. The research component describes a prospective investigation of endocrine-disrupting chemical (EDC) levels in maternal urine collected during pregnancy and relation to methylation and imprinting status of growth factors and methylation status of the ERa gene in cord blood from offspring. It is hypothesized that intrauterine exposure to synthetic EDCs including, bisphenol A and the phthalates di-(2-ethylhexyl) phthalate (DEHP), dibutyl phthalate (DBF), and butyl benzyl phthalate (BBP), present at significant levels in the environment, induces loss of imprinting of insulin-like growth factor 2 and hypomethylation of the imprinting control region of the LIT1 gene on chromosome 11p15. Further it is hypothesized that intrauterine exposure to bisphenol A and the phthalates DEHP, DBP, and BBP induces hypomethylation of the ERa gene and thereby increases ERa transcription expression. Urine samples of pregnant women will be collected and EDC levels will be measured at the Centers for Disease Control. Cord blood samples will be collected at delivery and DNA methylation and imprinting status will be determined. Features of this design include state-of-the art exposure ascertainment, sufficient statistical power to detect exposure-related variation in epigenetic control of gene expression, and efficient control for potential confounding variables.
DESCRIPTION (provided by applicant):
This award aims to extend the candidate's growing expertise in socioeconomic dimensions in housing as it relates to health disparities focusing in particular on childhood asthma risk. The candidate completed a fellowship in academic pediatrics, earning an MPH from Boston University School of Public Health in epidemiology, biostatistics and environmental health. Further research training will occur under the supervision of a multi-disciplinary Advisory Committee with expertise in asthma, life stress, housing, allergens, immunology, and qualitative research. Career development will be enhanced through course work and participation in seminars and research on stress and housing factors. This award, along with the candidate's clinical and scientific background, will facilitate the development of a successful independent research career with a focus on the epidemiological study of the influence of both social and physical dimensions of housing conditions on childhood asthma morbidity in the inner city. While many epidemiological studies describe associations between poor housing and health (including asthma), none have considered the psychological dimension of housing quality either independently or in combination with physical housing characteristics as related to asthma risk. The investigators hypothesize that children exposed to higher level housing stress will be at greater risk for asthma/wheeze syndromes, independent of other life stressors (e.g., neighborhood disadvantage or violence) and exposures associated with poor housing (e.g., cockroach or rodent). Furthermore, they hypothesize higher-level housing stress, assessed annually in the first three years of life, will predict subsequent increased total IgE levels and allergen-specific lymphocyte proliferative response in the child at age 3 years controlled for these other risk factors. Study of the relative importance of housing stress in urban populations may result in more targeted housing interventions and policy change to impact public health.
DESCRIPTION (provided by applicant)
Problem and Pilot Data: Environmental exposures during pregnancy can influence the future health of the developing child, but specific mechanisms are unknown. Pilot data show that offspring of mother mice exposed during pregnancy to diesel exhaust particles are more susceptible to allergy than babies of normal mothers. Remarkably, this effect is also caused by control, "inert" titanium dioxide particles. Moreover, '"inert" particles are actually pathogenic/pro-inflammatory in pregnant mice (while causing minimal effects in non pregnant controls). Hence, pregnancy causes a heretofore unrecognized change in lung responses to environmental agents, prompting the central hypothesis: pregnancy-related hormones alter local lung innate immune responses to inhaled environmental particles, leading to systemic cytokine modulation of developing offspring immunity to a state of greater susceptibility to allergy.
Specific aim 1: To characterize the local pulmonary and systemic response of pregnant mice to environmental particles, testing the prediction of enhanced pro-Th2 responses in BAL analysis, multiplex cytokine assays of serum and lung gene expression profiling.
Specific aim 2: To characterize effects of pregnancy hormones on the alveolar macrophage (AM) response to particles, testing the prediction that progesterone and estrogen promote pro-Th2 AM responses to particles in vitro and in vivo. Specific aim 3: To test the causal role of Th2 cytokines in particle-mediated effects during pregnancy by neutralizing antibody and cytokine treatment strategies.
Significance: Rigorous mentorship (aim 1) will prepare the candidate for transition to independence (Aims 2 and 3) and continue developing his research career in environmental disease at a faculty level. This research will provide novel insights into the basic question of early life origins of asthma and the applied question of how environmental agents mediate in utero effects.
This competitive renewal application for continued support of the Undergraduate Minority Summer Internship Program affiliated with the graduate Training Program in Environmental Health Sciences (T32- ES07155) represents the continuation of an extremely successful program targeting undergraduate students for exposure to environmental health sciences. The short term goal of our summer internship program is to support research training in a number of diverse areas relevant to environmental health intended for undergraduate minority students. We have, as our long-term goal, the attraction of minority scientists into graduate programs and careers in the environmental health sciences. Our proposal is to support minority students in our summer long laboratory based program. Since our last competing renewal we have enlarged the program to include 16 applicants per year who complete a 10 - week summer research internship under
the supervision of faculty members who are also members of the Biological Science in Public Health (BPH) program at the Harvard School of Public Health. The BPH program has co-sponsored this highly successful internship since 1989 with an exclusive targeting of minority students since 1993. Faculty who are preceptors on the graduate level Training Program in Environmental Health Sciences join the larger group of preceptors within the BPH program to act as preceptors for summer interns. Up to 269 applicants for the 16 positions are rigorously screened and the admitted students select their areas of research from material provided before arrival and after two days of oral presentation on potential research opportunities. Research
areas include environmental toxicology; mutagenesis; DMA damage and repair; cell cycle progression; signal transduction; gene regulation; molecular epidemiology; environmental assessment. The environment is one that is extremely rich and during the 10 - week internship the summer minority interns are specifically targeted for inclusion in a number of intern program related events that are rich in their scientific content.
DESCRIPTION (provided by applicant)
This is a competitive renewal of a highly successful short term training program in Biostatistics at the Harvard School of Public Health. This program targets undergraduate students who are members of groups who are underrepresented in the field of Public Health, including, primarily minority groups, and who are pursuing an undergraduate degree in mathematics, computer science, physics, engineering or another quantitative field. Support is requested for six 4-week internship slots. Among the activities supported by the program are introductory coursework in biostatistics and statistical computing, research seminars, small group research projects, career planning workshops, a practice GRE examination and a variety of social activities. Group projects will be designed each year in collaboration with faculty from the Departments of Biostatistics, Epidemiology, Society, Human Development and Health, and Environmental Health at the Harvard School of Public Health. The project will include data analysis, report writing, and oral presentation. All students who participate in the program will be strongly encouraged to go on to graduate school. While the emphasis will be on the fields of biostatistics and epidemiology, students will learn about a variety of research career options, including various aspects of environmental science. Promising students will be encouraged to apply to the investigators' School and will be given strong consideration for support on this training grant in Environmental Biostatistics (5-T32-ES07142) or on the other training grants in cancer and AIDS, for example. The progress over the past five years has been excellent. Sixty- students have participated in the program since 1994. Of the 43 students in the program who have finished their undergraduate degrees as of this time, 26 (62%) have gone on to pursue graduate training in statistics or biostatistics. Ten have entered the Biostatistics program and Harvard. The program is already having a measurable positive impact on recruitment and retention in the Department.
DESCRIPTION (provided by applicant)
The mission of the Training Program in Environmental Epidemiology is to prepare a cadre of outstanding researchers in environmental epidemiology through a program of rigorous training and research methodology. This Program is currently in its 24th year and is the established cornerstone of support for students working in areas of environmental and occupational epidemiology at the Harvard School of Public Health. To date it has graduated 140 professionals who conduct research, teach, and consult in environmental epidemiology and related disciplines, with 27 supported since the last competitive submission. The track in environmental molecular epidemiology, which was developed to provide trainees with comprehensive exposure to the application of molecular genetics techniques to environmental epidemiologic research, is now established and flourishing with trainees and new faculty. Training for both of these tracks is offered at the pre- and postdoctoral levels. Predoctoral students in the environmental and occupational track take courses in epidemiology, exposure assessment, biostatistics, occupational health, toxicology, biomarkers, molecular biology, and genetics. Additional research credits are obtained on the methodology of epidemiologic research through tutorials on current problems in epidemiology, and from the completion of a doctoral thesis of substance. In addition to course offerings, the track in environmental molecular epidemiology offers trainees the opportunity to select from four laboratories dedicated to research in molecular epidemiology with emphasis upon environmental and occupational diseases and cancer. The predoctoral training period usually lasts between 3 to 5 years, but varies depending on the students' preparation. The first 2 years are devoted primarily to course work. Most didactic teaching is shared by the Departments of Epidemiology and Environmental Health at the School of Public Health, but also includes a large host of affiliates working within the Harvard Medical area. Postdoctoral trainees are exposed to a wide variety of intellectual offerings, including didactic instruction, seminars, professional colloquia and meetings, and academic collaboration. Significant research usually begins for the postdoctoral fellow in his/her second year and varies in duration depending on the scope of work. The expansion of this Program to include trainees in the field of environmental molecular epidemiology has not only enhanced the Program's breadth, but defines the mission for the 21st century. This Program will continue a long history of excellence in training professionals in the field of environmental epidemiology at Harvard University.
DESCRIPTION (provided by applicant)
Boston University School of Public Health's Department of Environmental Health is one of a small number of academic units nationally that has a specialty in the application of environmental epidemiology in community settings. This proposal describes a training regimen, located within a mature doctoral program, whose ultimate objective is to prepare pre-doctoral students to use the most sophisticated methods currently available to advance the state of knowledge and develop new methods in community-oriented environmental epidemiology. While the department is primarily committed to advancing the research agenda by use of cutting edge tools (such as functional magnetic resonance imaging), development of new methods (such as dynamical systems models or lattice formulations) and integration of other disciplines (mathematics, urban studies, economics, toxicology and anthropology, among others), it also recognizes that even the most arcane scientific research takes place within a specific social context. In the case of environmental epidemiology this includes a complex mix of federal, state and local agencies, individual and organized community members and elements of the private sector. Students must be prepared to function in this difficult professional landscape. Accordingly, instruction about the social context is an integral part of the program. The program is designed to admit two students per year for a steady-state census of six predoctoral trainees. It consists of a stipulated core curriculum that includes instruction in epidemiology, toxicology, and biostatistics, followed by a suite of more advanced courses to accommodate various research interests and strategies. It also includes activities to enrich the experience of the trainees and promote social and professional cohesion.
DESCRIPTION (provided by applicant)
The Interdisciplinary Training in Genes and the Environment program at the Harvard School of Public Health (HSPH) will address the critical need for well-trained scientists who have an understanding of, and commitment to, cutting-edge research at the intersection of molecular and environmental exposure biology, and statistical and computational methods. The training program will involve active participation by 30 accomplished and experienced multidisciplinary faculty members, including environmental health scientists, molecular biologists, molecular epidemiologists, computational biologists, biostatisticians and bioinformaticians. The two interrelated goals of our proposed training program are: To train true collaborative partners able to pursue methodological research that is motivated by, and helps to solve, difficult analytic issues that arise in studies of human environmental exposures and genetic susceptibility to complex diseases; To encourage interdisciplinary research, especially in genetics and the various "omics" arising from new methodologies for characterizing biological activity associated with environmental exposures in laboratory and population sciences. Trainees will be pre-doctoral students and post-doctoral fellows at HSPH in the Departments of Environmental Health, Epidemiology, Biostatistics, and Genetics and Complex Diseases, which will jointly administer the grant. The program proposes initial support for 3 pre-doctoral students with 2 additional trainees in year 2 and 3 additional trainees in year 3 to bring an annual total of 8 pre-doctoral students into the program (year 3-8). The program also plans to support training of one post-doctoral fellow per year. These post-doctoral trainees will have an advanced degree (PhD, MD, MD-PhD or other doctoral degree) relevant to exposure biology/environmental health sciences or computational areas of genomics/proteomics and will have a specific interest in cross-training research experience. All trainees in the Interdisciplinary Training in Genes and Environment program will be provided an outstanding opportunity to become equally skilled in genomics, environmental health sciences and quantitative methods in order to attain leadership roles in interdisciplinary studies of human genes and the environment, with the ultimate goal of serving public health interests in developing effective disease prevention and intervention strategies.
DESCRIPTION (provided by applicant): The field of nanotechnology is exploding. The potential applications that exploit unique properties of common materials at the nanoscale are innumerable, and scientists are exploring these possibilities with increasing eagerness. Consequently, production of nanomaterials is outpacing the current understanding of their health effects. This study will help charaterize the toxicity of nanomaterials, so that the health of the public can be properly safeguarded against exposure to potentially harmful nanomaterials. The goal of this research is to address the uncertainty of nanomaterial toxicity by determining the effect of size, morphology, and surface charge on the bioavailability and toxicity of engineered nanomaterials while also revealing the cellular mechanisms that control their gastrointestinal uptake and intracellular processing. The specific aims of this study are: 1) determine the influence of particle size, shape, and surface change on the transepithelial transport eficiency, 2) determine the intracellular fate of engineered nanomaterials and the cellular response triggered by nanomaterial uptake, and 3) determine the influence of nanomaterial exposure on cell viability and the structural integrity of a mono-layer of epithelial cells. To acheive these aims, an in vitro gut model will be used to measure the transport efficiency of a library of nanomaterials ranging in size, morphology, and surface charge across a monolayer of epithelial cells, followed by real time tracking of single nanoparticles exposed to the same cell type and examination of exposed cell integrity by fluorescence microscopy. By applying these techniques systematically to nanomaterials of varying characteristics, a picture will emerge not only of which individual nanomaterials are more or less toxic, but also what properties are common to more and less toxic nanomaterials. With this knowledge, products and substances containing nanomaterials developed for applications involving widespread exposure to the population can be developed to minimize toxicity and the associated health risks. This applies directly to the goals of the National Toxicology Program of which the National Institute of Environmental Health Sciences of the National Institutes of Health is a core agency.
DESCRIPTION (provided by applicant):
This is a competitive renewal application for the Training Program in Environmental Health Sciences that is based at the Harvard School of Public Health. This is an interdisciplinary and interdepartmental program that currently supports 12 predoctoral and 3 postdoctoral trainees. Twenty-three Harvard University faculty members are listed as preceptors; most of these are in the Biology in Public Health (BPH) program at the Harvard School of Public Health (HSPH) and are members of the HSPH NIEHS Center for Environmental Health (an interdisciplinary unit that fosters research and administers the NIEHS Center Grant ES000002); the remaining preceptors are in the Biological and Biomedical Sciences (BBS) program in the Harvard Medical area. The faculty preceptors are located in the following departments: Cancer Cell Biology, Environmental Health, Nutrition, Epidemiology, immunology and Infectious Disease, Chemistry and Molecular Pharmacology, Microbiology and Molecular Genetics, and Pathology. The home base of this training program is the School of Public Health where there is a strong emphasis on the application of knowledge gained through basic research to the protection of human health from the effects of toxic environmental agents. Its strength derives from this fact as well as the fact that the training faculty has a broad range of skills ranging from structure/function analyses of environmentally relevant macromolecules (e.g., DNA repair and recombination proteins), to molecular epidemiological studies of human gene-environment interactions, to assessments of real-life exposures of human populations to environmental pollutants. The investigators continue to offer an array of basic studies on how bacteria, yeast, cultured mammalian cells, knockout and transgenic mice, and humans respond to environmental agents with emphasis on how the agents are perceived, how signals are transduced to various parts of the cell or animal, and on the biological consequences of toxin exposure. They believe that this program offers an exciting range of research for both predoctoral and postdoctoral training. The proposed training program is also proud of the superb quality of the applicants, the participating departments and the laboratories of individual preceptors. A rigorous training is proposed for both predoctoral and postdoctoral trainees, and it is expected that this training will position the trainees for productive careers in the Environmental Health Sciences in academia, government, etc.
DESCRIPTION (provided by applicant)
This is a request for an extension of the training program in Environmental Health Statistics at the Harvard School of Public Health. The program prepares pre-doctoral and postdoctoral fellows for research in the application of biostatistics to environmental health. The program will be administered through the Department of Biostatistics, with active participation by faculty members from the Department of Environmental Health and Department of Epidemiology, also located at the Harvard School of Public Health. Trainees will receive high-quality instruction in basic biostatistical theory and methods, such as probability, statistical inference, computing and data analysis. The program will also provide training in specialized topics of particular relevance for environmental applications, such as longitudinal analysis, missing data techniques, statistical genetics and high-dimensional data analysis, and statistical methods relevant to environmental exposure assessment such as measurement error models and spatial statistics. Training will also be provided through applied course work in environmental health and a regular seminar series called "Environmental Statistics Seminar Series", where faculty, students, and fellows present their own environmental health-related research. An important focus of training will be the opportunity to collaborate with faculty members on biostatistical research as it applies to environmental health. All trainees will participate in Harvard's program on scientific integrity in the conduct of research as well as professional development workshops focusing on strategies for success in academic settings.
Since its inception in 1982, this training program has emphasized strong links to the environmental sciences. In recent years, program trainers have placed particular importance on the recruitment of students from underrepresented minority groups. The focus on interdisciplinary training at the Harvard School of Public Health, as well as its talented and diverse student body and faculty, makes it ideally' suited for a training program in Environmental Statistics.
BACKGROUND
This is a competing continuation application for a Training Program in Environmental Statistics at the Harvard School of Public Health. The program was started in 1982 and has been continuously supported. The emphasis of program is to train biostatisticians and other quantitative scientists in the theory and practice of statistical science with application to environmental health research. The application is justified based on the increasing need to analyze complex data in environmental health that resulted from the recent advances in data collection technology, and significant interaction of the training program with other funded research projects in environmental health.
New over the past five-year funding cycle are two features. In 2003, the Department of Biostatistics entered into an arrangement with the Statistics Department and the Graduate School of Arts and Sciences (GSAS) at Harvard to offer the Ph.D. instead of the S.D. offered by the School of Public Health. This impacts the Training Program by making available the recruiting efforts from the GSAS for qualified Ph.D. candidates; students now qualify for additional scholarships available only to Harvard Ph.D. students (important for under-represented minorities); and this change improves the availability of research mentoring from GSAS faculty in Statistics.
Additionally, in 2003 when Dr. Ryan was on sabbatical, Dr. Coull took over as Director of the program. Since her return, Dr. Ryan has been named the co-Director. In 2005, two new faculty members were hired with research interests in environmental statistics. The Training Program is currently administered using an "Executive Committee approach" (Dr. Coull as Director, Drs. Ryan and Lin serve as co-Directors, with Dr. Paciorek "involved in all administrative decisions").
DESCRIPTION (provided by applicant): Parkinson's disease (PD) is the second most common neurodegenerative disease, affecting approximately 1% of the population older than 65. Recent research has identified a metal ion transporter, ATP13A2, that that when mutated leads to early-onset PD. The Lindquist lab has found that the Saccharomyces cerevisiae homolog of this protein, YOR291 wp, suppresses the toxicity of another PD protein, ?-synuclein. The goal of this proposal is to understand the biological function of ATP13A2/YOR291wp and the role metal ion transport plays in ?-synuclein toxicity via the following specific aims: 1) Characterize YOR291w function using molecular genetic and biochemical approaches 2) Investigate the effects of metal ion transporters on alpha-synuclein toxicity using high-throughput screening and microarrays and 3) confirm all results in neuronal PC12 cell line. The results from these experiments will identify the biological function of ATP13A2/YOR291wp, as well as the role metal ions play in ?-synuclein toxicity and the development of PD. Public Health statement: Parkinson's disease is a common neurodegenerative disease. I will investigate the function of a protein shown to cause familial Parkinson's disease, as well as the role metal ions play in the development of Parkinson's disease. Understanding how Parkinson's disease progresses will lead to the identification of new therapeutic targets.
DESCRIPTION (provided by applicant): Acute respiratory infection (ARI) is responsible for 19% of all deaths worldwide among children less than five years old. Environmental factors such as air pollution have been hypothesized to increase the risk of incidence for ARI in young children. Malnutrition is also known to suppress the immune system that renders a child more
susceptible to ARI. This five-year traineeship aims to link environmental pollutants, while controlling for host nutritional factors, to an overall increase in the incidence of respiratory diseases in Ecuadorian children. Air pollution in Quito, Ecuador will be closely monitored by the City of Quito and using mobile handheld monitors to allow a time-series and geographical analysis of respiratory disease in a cohort of children already enrolled in an NIH-funded study. These data will be used to statistically analyze adverse respiratory health outcomes in variably
malnourished Ecuadorian children as it relates to air pollution. In addition, molecular and microbiologic analysis will establish a spectrum of pathogens that cause disease.
DESCRIPTION (provided by applicant)
This training program in environmental toxicology produces scientists professionally qualified to make original contributions directed at improved understanding of the impacts of hazardous chemicals and other environmental agents on human health. Emphasis is placed on development and application of in vivo and in vitro experimental models designed ultimately to elucidate, in cellular and molecular terms, mechanisms through which environmental and endogenous agents induce their adverse effects. These approaches help to identify health hazards resulting from the presence of toxic agents in the environment; moreover, the identification of biomarkers of exposure provides benchmarks for assessing the effectiveness of intervention measures put in place to reduce human risks. Strong emphasis is placed on the development, validation and application of methodology for detection and characterization of adverse effects. These methodologies improve assessment of actual or potential hazards to humans resulting from environmental exposures. Utilizing biochemical, chemical and biological approaches, the training of doctoral candidates and postdoctoral trainees is concerned with: characterization of the effects of toxic, carcinogenic and mutagenic chemicals at the intact animal, tissue, cellular and molecular levels; development of methods for the detection and quantification of such effects in humans, animals, and other experimental systems; studies of metabolic activation, macromolecular binding and genetic effects; and, elucidation of modes of action at cellular and molecular levels. Training involves didactic courses, formal and informal seminars and other means of information exchange, and laboratory research projects utilizing appropriate model systems for problem definition and solution.
DESCRIPTION (provided by applicant)
TNEC is applying to the HWWT program. The primary goal is to provide hazardous waste worker and/or emergency responder health and safety (H&S) training to 800 workers per year for five years in five New England states (MA, CT, RI, NH, VT). The project will train 40% of workers in multiple-day courses (24-hour Emergency Responder or 40-hour Hazardous Waste Site Personnel). TNEC will train 60% percent of workers in Refresher, Supervisor / Incident Commander / NIMS, or Awareness Level ER training, including Hazard Disaster Preparedness training related to chemical, biological, radiation/nuclear and explosive agents (CBRNE) (all 8-hour programs). The intended purpose is to support the implementation of HAZWOPER H&S programs to prevent occupational illnesses, injuries and fatalities, including during emergency response to accidental or deliberate releases of CBRNE agents that could adversely affect workers and neighboring communities. The participatory design of the training, uses empowerment-oriented worker education methodology, and encourages trainees to practice, workplace H&S decision-making.
Specific Aim 2 is to continue and expand several ongoing initiatives by strengthening outreach and marketing efforts to increase its client base of 2,190 private and public sector firms and organizations. It will include collaboration with the OSHA Education Center at Keene State University and the UML Center for Industrial Competitiveness. TNEC will expand partnerships with state agencies and municipalities in need of "all hazards" H&S training for emergency response, public health, and criminal justice personnel. Secondly, the applicant will continue to utilize the integration of existing advanced training technology based training modules and maintain the integrity of hands-on and small group activities with computer and web-based training. These modules will incorporated into emergency response training courses. TNEC intends to incorporate web-based modules for in-class and after-training access, through a partnership with Y-Stress, Inc., a current NIEHS WETP SBIR program awardee. The applicant will also continue to provide 40-hour HAZWOPER training courses for Minority Worker Training Programs, that target low-income young adults from underserved, urban communities, and help to increase their workforce and the employment opportunities.
Specific Aim 3 is to sustain the capacity of TNEC member organizations to lead and support the regional and national H&S movement by supporting and strengthening the capacity of the COSH organizations in Connecticut, Massachusetts, New Hampshire and Rhode Island as essential resources for high quality, technically accurate, and educationally appropriate worker-oriented safety and health training, and further enabling them to provide health and safety training to hazardous waste/emergency response workers; continue to strengthen U MASS Lowell's worker H&S education capacities; and continue to build strong links among public health professionals, worker health educators, and labor movement health and safety activists in order to maintain high quality training for hazardous waste workers and emergency responders.
DESCRIPTION (provided by applicant)
Little capacity exists at the community level for addressing and improving the occupational health status of workers. Within immigrant communities, the lack of access to occupational health information (prevention, treatment and benefits) and basic medical care is complicated by marginal economic stability and employment in small immigrant-owned or family-owned businesses. Ethnic networks, business owner networks, and existing links to community organizations, including community health centers, represent an opportunity to reach low-income immigrant workers with culturally appropriate occupational health programs. The Dorchester Environmental Health Coalition (DEHC), representing three community health centers, ethnic community organizations, the Massachusetts Coalition for Occupational Safety and Health, block and neighborhood associations and youth groups, has taken action on assessing environmental health needs of Dorchester residents. DEHC is proposing to bring three community health centers, several community organizations and the Harvard School of Public Health, the Chief Medical Officer of Dorchester House to engage the community in developing and implementing community-based occupational health programs among immigrant groups in Dorchester, Massachusetts.
The purpose of the proposed project is to: assess the occupational health needs of low-income and immigrant communities in Dorchester; involve workers, their families and small businesses in designing culturally appropriate interventions; carry out community education; and evaluate the impact of these efforts on the Dorchester community as well as local policy and regulatory functions. The specific aims include: 1) Conduct a streamlined assessment of the occupational health needs of Dorchester's low-income and immigrant communities, augmenting available current statistical data and the results of recent resident surveys with focus groups, key informant interviews, and culturally acceptable methods of obtaining information; 2) Develop culturally appropriate models of education, communication and dissemination of occupational health information - based on research, intervention and policy objectives - within the Dorchester community with an initial focus on two populous, undeserved groups: the Vietnamese and Cape Verdean communities; 3) Implement community education and training programs, and conduct demonstration projects to achieve a demonstrable improvement in access to occupational safety and health information and resources within the Dorchester community; and 4) Provide site-specific feedback and technical assistance to small businesses and community health centers to increase awareness of the occupational health needs and to promote local prevention activities in small business and community settings.
Crisp Terms/Key Words: health education, southeast Asia, community health service, health care service availability, health economics, information dissemination, occupational health /safety, health survey, low socioeconomic status, culture, work site, immigrant, health care policy, behavioral /social science research tag, health services research tag, educational resource design /development, medically underserved population, health disparity
DESCRIPTION (provided by applicant)
Low-income people living in depressed neighborhoods of Worcester, MA, are disproportionately exposed to environmental stressors: 1) a major toxic chemical pollution burden; 2) unhealthy physical features like numerous brownflelds and lack of greenspace, and 3) pervasive social violence and insecurity. Compounding their vulnerability is limited adaptive capacity rooted in socio-political and economic stress. Approaches that focus on single problems will be ineffective. Our working groups of local community representatives, researchers, city health and environment agencies, the youth center and a local NGO plan an inclusive, systems-based approach to improve adaptive capabilities in Worcester's Piedmont and Main South Neighborhoods. Worcester was once the heartland of the American Industrial Revolution, and its built environment now bears a significant historical and ongoing pollution burden. Similar conditions are found in many medium-sized cities across industrial America. The project has four stages, all neighborhood-based: 1) Strategic Assessment - detailed descriptions of the baseline; 2) Strategic Planning - identification of priority stressors and opportunities; 3) Implementation - making priority interventions and developing capacity; and 4) Performance Monitoring - measurement to detect significant changes (post- vs. pre-policy values). Products will include: neighborhood centered databases, planning documents, and evaluation reports; a practitioner's manual; and research reports of observations and findings. Two hypotheses are tested: a) Primary built environment stressors of a physical, chemical and socio-economic nature conspire together to create vulnerability in Worcester's Main South and Piedmont Neighborhoods; and b) this vulnerability system can be described and improved through a participatory process that fosters experiential learning, builds community ownership, strengthens adaptive capacity of those at risk, and makes environmental and health promotion policies responsive to those most in need.
Crisp Terms/Key Words: environmental exposure, cost effectiveness, clinical research, health care policy, quality of life, environmental stressor, politics, community, socioeconomics, experience, social model, model design /development, interview, human subject, health care cost /financing, public health, human ecology, disease /disorder proneness /risk
DESCRIPTION (provided by applicant)
This project links breast cancer and environmental justice advocacy through a variety of approaches conducted by Silent Spring Institute, a community-based breast cancer research organization; Communities for a Better Environment, an environmental justice organization and partners from Brown University. The project will conduct home environmental exposure assessments, collecting air and dust samples to assess indoor levels of pollutants, especially endocrine disruptors that are potentially linked to breast cancer, reproductive and neurological development, and other health outcomes. Data collection, analysis, community education, and organizational linkages will occur in two locations - Cape Cod, Massachusetts, a region of unexplained high breast cancer incidences that has been the focus of prior work of Silent Spring Institute, and a new site in Richmond, California, that is largely composed of people of color and impacted by industrial facilities. Study results will be shared both as aggregate information presented through community meetings, news media, and on-line, and as individual report-back information to study participants. Using both report-back approaches, this study will seek to maximize understanding of exposure data and its limitations, and address the ethical issues of ensuring community and individual autonomy, the right to know, and ultimately the right to act on scientific information by reducing exposures. Specific aims for this project include: 1) Conduct community-based exposure assessment research, collecting dust and air samples to assess levels and sources of pollutants in homes located in Cape Cod and Richmond; 2) Link breast cancer and environmental justice advocacy through community-based outreach, including public meetings, web sites, and publications; 3) Develop ethical methods for reporting back study results to communities and individual study participants, and produce a handbook for general use in reporting results of community exposure studies; 4) Pilot test an intervention to reduce household pollutant levels.
Crisp Terms/Key Words: geographic difference, biohazard detection, environmental exposure, hazardous substance, clinical research, Internet, method development, handbook, dust, breast neoplasm, environment related neoplasm /cancer, biohazard control, interview, data collection, information dissemination, human subject, community health service, ethics, endocrine disorder, health education, consumer product, air sampling /monitoring, air pollution
We propose to establish a Center for Children's Environmental Health and Disease Prevention Research that addresses the concerns of a community living in the Tar Creek Superfund site of Oklahoma - an area highly contaminated by metals (lead, cadmium, iron, manganese, and others) in mining waste and populated by many residents of Native American descent. Our overall goal is to take a highly innovative and integrated approach to addressing a "real world" problem, i.e., the potential of the mixtures of metals that are present in "chat" (mining waste) to interact with each other in terms of exposure, absorption, dose, and adverse effects on the development of children.
Our Center will pursue four Research Projects with the support of four Cores. Project 1 will be a community-based participatory epidemiologic study that examines biological markers of fetal and early childhood exposure to metals (lead, manganese, cadmium, and iron), their impact on measures of mental development, and their response to a quasi-experimental randomized trial
of nutritional and behavioral interventions. Project 2 will assess the utility of size fractionation and sequential extraction studies for characterizing chat, conduct a nested case-control study of the determinants of high versus low burdens of metals amongst children participating in Project 1, and produce standardized "homogenized chat" for Projects 3 and 4. Project 3 will investigate the expression of binding and transporter molecules for metal transport and the corresponding pharmacokinetics of metals from the lung and gut to the blood, CNS and other organs as they relate to pregnant rats and their weanlings. Project 4 will examine the
effect of pre- and neo-natal exposure to metals on neurochemical changes and neurobehavioral outcomes in rats. The effect of simple mixtures of metals will be compared with the effect of "homogenized chat" in both Projects 3 and 4. The potential effect of stress from living near toxic waste will be explored in Project 1 and the potential modifying effect of stress on metals
neurotoxicity will also be explored in Project 4. Our Administrative, Analytical Chemistry, and Biostatistics Cores will enable us to fully integrate and support our research, and our Community Outreach and Translation Core will utilize an innovative portfolio of outreach activities developed in conjunction with a broadly-based Community Advisory Board to develop awareness and
influence behaviors and health practices in order to prevent adverse health effects in children from exposure to metals in mining waste.
Crisp Terms/Key Words: environmental exposure, clinical research, mining, metal poisoning, human subject, environmental health
DESCRIPTION (provided by applicant): The proposed work will develop nanoscale tools for characterizing the mammalian cell; it will ultimately lead to new tools for drug discovery, diagnosis of disease, and studying fundamental cell biology. Its justification is that study of biological entities fundamentally involves the study of nanoscale components of the cell: subcellular organelles, pathogens, macromolecules. Nanoscale tools are required to examine and analyze these components at the subcellular scale. The research will create nanometer-scale components (rods, particles, and surfaces) using "biology-friendly" nanotechnology (soft lithography and self-assembled monolayers), and use them to examine mammalian cells. It will use nanoscience-based approaches to:1) create 2D and 3D microenvironments with controlled shapes, molecular composition, and mechanical characteristics for studies of cells; 2) create electrically, optically, and mechanically functional nanosystems that permit selective stimulation of cells, and allow read-out of cellular electrical, chemical and mechanical responses with subcellular resolution; 3) leverage systems that exhibit quantum phenomena unique to nanosystems (e.g., superparamagnetism, superluminosity) to generate new physics and chemistry relevant to biology, and use this understanding of physical science to afford fundamentally new classes of information about cell structure and function; 4) develop methods to multiplex nanoscale technologies to measure functions and characteristics of single cells in parallel, with high statistical reliability; 5) demonstrate the relevance and application of these tools using important biological problems. The work will combine to generate a "nanotool cellular workbench"; it has four specific aims: 1) To create novel multifunctional nanometer-scale structures, particles, components, and surfaces, and analytical systems that use these entities, 2) To use this "work bench" of nanotools to understand how individual cells sense mechanical cues and integrate them with chemical and electrical signals in 2D and 3D microenvironments, 3) To create nanoscale control interfaces that rapidly actuate changes in cellular signal transduction and read-out biochemical responses, and 4) To combine these nanotechnologies with microfluidic systems to create prototypes for integrated cellular biochip-based medical devices.
DESCRIPTION (provided by applicant): Organophosphorus (OP) pesticide poisoning is a leading cause of premature death in many developing countries, killing an estimated 200,000 people every year in the Asia-Pacific region alone. In North America and Europe, the situation is quite different. While pesticide poisoning does occur, the main risk of OP poisoning is from terrorist attacks on civilian populations - through the release of OP nerve gases in crowded spaces or perhaps introduction of highly toxic pesticides into water supplies. The acute toxicity of OPs is primarily due to inhibition of acetylcholinesterase (AChE). Current therapy for OP poisoning requires resuscitation and use of atropine, followed by administration of oximes to reactivate AChE. However, these antidotes have limited effectiveness and between 10 and 40% of patients, depending on the responsible OP, still die even with intensive care support. Although OP pesticides have been a clinical problem for 50 years, no new therapies have been introduced since the 1960s. Because early therapeutic interventions lead to improved outcomes after OP poisoning, a treatment that is safe and highly effective, and that can be given by first responders at the site of poisoning, should markedly improve outcome. Both bacteria and humans make enzymes that hydrolyze OP compounds. Recombinant bacterial OP hydrolases have the potential to provide an affordable, widely available, and safe treatment that is rapidly effective against a wide variety of OPs. CSIRO, Entomology, in Australia has developed a bacterial enzyme, called OpdA, with excellent in vitro catalytic activity against many currently used OPs. In proof-of-concept studies, we have shown that OpdA has excellent efficacy when used alone or with 2- PAM in rat models of parathion and dichlorvos poisoning. However, a number of further steps, including the proposed non-human primate studies, are required before clinical trials in humans with OP poisoning. The purposes of this grant are to develop a new non-human primate (NHP) model of parathion poisoning and to test the safety and efficacy of OpdA in this NHP model. Proof that the enzyme is safe and effective against parathion should provide the necessary impetus for further development for human use. Our central hypothesis is that OpdA is safe and improves survival after poisoning with parathion. If successful, the proposed research promises to improve public health by mitigating the acute toxic effects of pesticides after accidental, intentional, or suicidal poisoning. The research also has implications for the treatment of military personnel and civilians after nerve gas poisoning.
DESCRIPTION (provided by applicant):
There are approximately 45 million current smokers and 46 million former smokers who are at increased risk for tobacco-related disease in the United States. The public health implications of this widespread environmental exposure are profound; tobacco smoke is the leading preventable cause of death in the United States and is projected to cause nearly 450 million deaths worldwide during the next 50 years. Despite the causal role of cigarette smoking in lung cancer and COPD, only 10-20% of smokers develop these diseases. There are few indicators of which smokers are at highest risk for disease, and it is unclear why individuals remain at high risk decades after they have stopped smoking. Current standard methods for quantifying exposure to tobacco smoke are limited in their ability to accurately assess cumulative dose and past exposure, and they do not capture the physiologic host response to tobacco exposure. We have previously shown that cigarette smoke causes an airway-wide epithelial cell "field of injury" and that gene expression, in airway epithelial cells obtained at bronchoscopy, reflects host response to smoking. We propose here to extend the "field of injury" concept to easily-accessible airway epithelial cells that can be obtained from nasal or buccal mucosa in a non-invasive fashion. By measuring global gene expression at these sites using a new "all-exon" expression platform, we will develop a series of biomarkers that assess host response to current tobacco exposure (active vs. passive vs. never smokers), intensity of current exposure, cumulative exposure among current smokers, time since last exposure among smokers who recently quit, and lifetime exposure. Furthermore, we will develop molecular pathway-based gene expression biomarkers that may be more accurate markers of individual responses to tobacco smoking. We also propose to correlate airway gene expression biomarkers with lung function and systemic markers of oxidative stress and inflammation, setting the stage for a more detailed understanding of how variability in epithelial response contributes to variability in disease-related pulmonary and systemic sequelae of tobacco smoke exposure. These studies will establish a new non-invasive tool that can be used to measure the host responses to tobacco smoke that can be used in subsequent large scale population studies as part of the Genes and Environment Initiative.
DESCRIPTION (provided by applicant):
DNA damage is an important risk factor for cancer and many other diseases. Whether induced by the environment or created endogenously, covalent modifications to DNA structure can be both cytotoxic and mutagenic. Being able to measure DNA damage and repair in human samples is therefore fundamentally valuable, both for delineating environmental conditions that render cells vulnerable to mutations, and for revealing genetic factors that modulate susceptibility to DNA damage. The single cell gel electrophoresis assay, or 'comet assay' is one of the most sensitive and versatile approaches for measuring DNA damage in human cells. It is grounded on a simple principle: when visualizing electrophoresed cells embedded in agarose, undamaged DNA is supercoiled and highly compact, whereas damaged DNA (relaxed loops and fragments) can more readily migrate, giving rise to the appearance of a bright nucleoid with a comet-like tail. Despite its proven efficacy, the comet assay is underutilized in studies of environmental risk factors in epidemiological studies, mostly because of a lack of standardization that has lead to inconsistent results among researchers and due to the time/labor required to perform the assay. Here, we propose to apply lab- on-a-chip technologies to create a "comet-chip" that will not only overcome problems in standardization, but will also permit high-throughput parallel processing of dozens of samples. We anticipate that the proposed technology will render this assay useful in a broad range of clinical, epidemiological, and experimental settings. In Specific Aim I, droplet-array methodology will be exploited for parallel processing of dozens of samples. Specific Aim II is aimed at combining cell patterning technologies with the comet assay in order to asses the impact of microenvironment on variability among cells. For Specific Aim III, we propose to create a self-contained comet device to provide much needed consistency. Finally, for Specific Aim IV, we propose to apply the 'comet chip' to mouse and human samples of varied DNA repair capacities, and to rigorously evaluate reproducibility and sensitivity. Importantly, while sophisticated equipment will be necessary for the highest-end processor that we propose to create, many of the proposed modifications can be applied to the comet assay in a fairly low-tech fashion, and thus can be widely disseminated. A high- throughput DNA damage and response sensor will be invaluable both for discerning dangerous environmental exposures, and for evaluating the efficacy of policy decisions aimed at reducing relevant exposures. It is hoped that the proposed technology will yield data to empower policy makers in the development of wise intervention strategies that will effectively prevent cancer and other illnesses, long before disease onset.
DESCRIPTION (provided by applicant):
Links between diet and human disease, and between reactive species and disease, are so commonly considered as to lie in the realm of textbooks and the popular press. Links between mitochondria and energy production are generally appreciated by junior high school. Links between mitochondria and calcium (including signaling), free radicals, or cell death may be less known to the general public, but each has in excess of 10,000 PubMed citations. However, despite broad and strong theoretical considerations supporting casual connections between diet effects on mitochondria and diet effects on disease - and some specific experimental support - there are, to our knowledge, no systematic studies that bridge this fundamental gap. Bridging this gap is central to understanding environment-gene interactions, as suboptimal dietary macronutrient choices are arguably the major environmental stressor in individuals living in Western societies. We therefore propose to bridge this gap using an interdisciplinary, product-development approach to discover and confirm innovative plasma metabolomic and proteomic biomarkers for dietary intake of subclasses of fats and carbohydrates, and for their effects on mitochondrial (dys)function. We will then validate these markers by using them to test the hypothesis that diet-associated effects on mitochondria are linked to diet-associated changes in disease risk. Five Aims are proposed. Aim 1 To determine the effects of dietary changes in fatty acid and carbohydrate composition on mitochondrial physiology Aim 2 To determine the effects of dietary changes in fatty acid and carbohydrate composition on the plasma metabolome and proteome Aims 3 and 4: To determine the extent to which adherence to/presence of each diet, dietary constituent, and mitochondrial property predict type II diabetes (Aim 3) and breast cancer (Aim 4) in previously profiled case control studies nested within the Nurses' Health Study Aim 5: To provide an electronic archive of the metabolomic and proteomic constituents of the blood of participants that could be repeatedly mined for future testing of new hypotheses. The proposed studies are directly responsive to the RFA and further general NIH goals of focusing on health and early interventions rather than late stage disease.
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