To fully understand the protective effect of pregnancy and also explore the possibility of recreating a parous-like stroma in a nulliparous individual as a preventive therapy, we must identify if the pregnancy effect is local (takes place in the MG) or systemic (the effect occurs at the hormonal level).

Common cell culture experiments are 2-dimensional (2D) lacking the 3D context of in vivo tissue and cannot be compared to the level of organization observed in the MG. We will use an in vivo-like 3D culture of mammary stromal-fibroblasts (a major cell type of the stroma involved in directing the stromal half of the dialogue with the epithelium to maintain tissue homeostasis). These cells form a 3D matrix resembling the tissue of origin.

We will use fibroblasts from parous and virgin rats to compare the matrix spatial organization and composition. We will test whether treating virgin fibroblasts with pregnancy levels of estrogen and progesterone produces matrices similar to those derived from parous fibroblasts.

Finally we will test the effects of parous and virgin fibroblast-derived matrices on the behavior of tumor and normal mammary epithelial cells. We will focus our attention on some of the most common tumor cell features: migration, invasion, proliferation and adherence to the matrix. We expect the biochemical and/or spatial organization of the parous and virgin fibroblast derived 3D matrices to be different with regard to repressing the tumor behavior.

The success of this project will allow us to test the identified putative “tumor repressor" molecules, through their signaling to the epithelium (soluble factors or physical forces). If we show that extracellular matrix composition and organization play important roles in supporting or repressing tumors, we may revise the current strategies used to prevent tumor formation.

PI: Malamy, Michael
Title: Genetic Systems to Study Virulence in Bacteroides
Abstract: This study will focus on factors that allow the obligate anaerobe Bacteroides fragilis, although a component of the normal colonic microbiota, to be a successful pathogen. These include its ability to withstand an aerobic environment (aero-tolerance) during early stages of infection; the presence of systems to import heme into the cell for the heme-dependent pathways of central metabolism and defense against reactive oxygen species; the ability of B. fragilis to remove sialic acid residues from host components, and its virtuosity in obtaining nutrients for growth in vivo from complex oligosaccharides and glycoproteins.

Specific aims include:

1. To continue to study factors that allow B. fragilis to withstand prolonged oxygen challenge (aerotolerance): We propose that activities in the B. fragilis periplasm serve as the initial line of defense to combat the formation of reactive oxygen species (ROS), protect sensitive targets from ROS challenges and to reverse ROS damage. In addition we have identified specific functions (superoxide dismutase, SOD), and an extensive gene cluster (the Bat operon) that are required for aerotolerance. We will test the hypothesis that the Bat operon forms a multi-protein complex in the cell membrane that plays an important role in exporting reducing potential from the cytoplasm to the periplasm.

2. Acquisition of iron and heme is important for B. fragilis growth in vitro and in vivo. We will study the process of heme uptake in B. fragilis by the heme permease systems whose genes and functions we have described We will also continue to study the heme-dependent, and Fe-S cluster-containing enzymes in the dual pathways of central metabolism to establish their roles in aerotolerance and in providing energy during oxygen challenge.

3. To investigate the composition, functions and control of operons for the acquisition of growth substrates from the infected host. We will focus on the operon containing the neuraminidase (nanH1) gene and several other glycohydrolases capable of converting the complex Lewis antigen found on the surface of many human cells to individual monosaccharides. We will continue to analyze the operon for NANA utilization, the NanLET operon and to define the sites in the three NanR repressed promoters for NanR binding. We will determine if neuraminidase is a virulence factor because it supplies NANA for growth, or because its activity alters the surface of host cells, or both.

PI: Mangili, Alexandra

Title: Postprandial Endothelial Dysfunction after a High-Fat Meal in HIV-Infected Men

Abstract: With access to HAART, HIV infection has become a chronic manageable illness and exposure to antiretroviral medications can extend over decades. At the current time, traditional risk factors appear to be the strongest predictors of the development of premature atherosclerosis in HIV-infected patients. To what extent selected antiretroviral medications or HIV infection itself contributes to the increased CV risk remains to clarify. Non-invasive markers of subclinical atherosclerosis are being introduced for early detection of CV disease in HIV-infected individuals and may improve the prognostic stratification of HIV-positive patients at risk for adverse CV events. Studying the progression of subclinical atherosclerosis by surrogate markers requires extended periods of follow-up. Endothelial dysfunction is an impaired vascular biologic state, manifests early in the development of atherosclerosis and can be studied by measuring ultrasonographic flow-mediated dilation (FMD) of the brachial artery. It has been previously demonstrated that endothelial activation can be induced by a single high-fat meal in the general population. HIV-infected patients are known to consume diets high in total and saturated fats, and do not meet the current American Heart Association dietary guidelines for the prevention of adverse CV events. The goal of this proposed pilot project is to examine the relationship between specific dietary factors and abnormal vascular function. Knowing how abnormal postprandial states contribute to subclinical atherosclerosis could ultimately lead to more effective dietary intervention strategies in HIV-infected patients and prevention of diet-related CV implications. The proposed pilot study will provide excellent potential for the development of other competitive application for funding in the cardiovascular and metabolic complications in HIV infection. It also will foster inter-disciplinary collaboration as the preparation of this grant involved specialists from the areas of Infectious Diseases, Nutrition and Cardiology.

PI: Mariner, Jeffrey
Title: Generic Lessons on Enabling Livestock Innovation in Chronic Environmental
Abstract: The two most common types of extended emergencies experienced by livestock keepers today, chronic environmental instability (CEI) and situations of chronic conflict and political instability (SCCPI), are dominated by iterative cycles of increasing vulnerability and disaster relief. SCCPI, characterized by violence and the persistence of long-term insecurity, is related to evolution of war economies where powerful players develop vested interests to continue conflict. Broadly defined, SCCPI include both open conflicts and low grade insecurity that preclude effective routine governance, institution building and development activities. Thus, SCCPI not only include conflicts such as southern Sudan, Somalia and the DRC but also portions of northern Kenya, southern Ethiopia and Uganda.

Likewise, land degradation, the most common cause of environmental instability, has its most devastating impacts in areas where changing land use patterns put livestock keepers at more risk for failure, forcing them to engage in production and environmental management practices that create increasing levels of vulnerability. 

In East Africa, almost as much land area falls under the domination of donor emergency policies as is served by development budgets. A major component of international aid funds repeated cycles of relief interventions with the objective of saving lives. These repeated cycles of relief interventions are suspected to increase poverty by disrupting livelihoods and increasing vulnerability and dependence. They destroy markets and decrease incentives for production.

The most recent environmental example in the news is the drought in Niger where food aid has started arriving after a successful harvest and threatens the livelihood of farmers. A recently documented example is the collapse of the sophisticated veterinary infrastructure and livestock sector in the Eastern DRC when long-term subsidies for pharmaceuticals disappeared. In East Africa, political and environmental instability often occur together and feed one another.

PI: Mason, Joel
Title: Doctoral Program in Human Nutrition
Abstract: The Postdoctoral Training Program in Human Nutrition and Metabolism combines outstanding resources for nutrition research with a uniquely qualified faculty of scientists in human nutrition to train physicians, other graduate-level health professionals, and selected PhDs to conduct research in human nutrition, metabolism, and the biological sciences basic to nutrition. Participating institutions include the U.S.D.A. Human Nutrition Research Center on Aging at Tufts University, Tufts-New England Medical Center, the Tufts University School of Nutrition Science and Policy, the Tufts University School of Medicine and the Sackler School of Graduate Biomedical Sciences. The Boston Obesity Research Center complements these institutional resources. There are 37 faculty members in this program 28 of who are preceptors and 9 of who are supporting faculty. The preceptors serve as principal investigators for over 20 NIH grants and have trained over 215 pre- and post-doctoral trainees over the last 15 years. Faculty research ranges from molecular and cellular biology to epidemiology and policy in areas of nutritional science relevant to the prevention and treatment of diseases such as cancer, heart disease, diabetes, hypertension, stroke and osteoporosis. The program primarily recruits physicians, although other graduate-level health professionals and selected PhDs are also recruited, and the program has actively sought out minority trainees. Renewal of four funded slots is requested. Trainees pursue an academically rigorous three year training program in which they have the additional options of obtaining either a PhD in nutrition or an M.A. in clinical care research. Progress in the program is monitored by a Steering Committee that includes the trainees' preceptors or thesis advisors. Research education is complemented by a rich schedule of research conferences and seminars and, for trainees with health professional degrees, by clinical training in an inpatient nutrition support service, and by rotation in outpatient nutrition subspecialty clinics. Trainees acquire a broad background in basic nutritional science and nutrition research, familiarity with the current literature in nutrition and disease, training in the use and interpretation of methodologies to assess the nutritional status of individuals and populations, and a sophisticated understanding of the applications and limitations of research design and biostatistical methodologies.

PI: Mason, Joel
Title: One-Carbon Nutrients in Cancer Prevention
Abstract: Accumulating evidence from many different types of scientific studies indicate that inadequate intake of the B vitamin, folate, increases the risk of several cancers. The evidence is most compelling for cancer of the colorectum. However, the prevailing evidence suggests that folate depletion only results in cancer when it operates in concert with other ill-defined genetic and environmental predispositions to cancer. The principle investigator of this K05 application has devoted the majority of his career to investigating this relationship between folate and carcinogenesis. He is now poised to begin to investigate how several other factors, such as age, tobacco, and vitamin B6 and B12 depletion interact with folate metabolism in such a way as to accentuate the promotion of cancer conveyed by folate depletion alone. The financial resources afforded by this K05 award will enable the investigator to dispense with a very substantial burden of clinical and administrative duties in his medical center, thereby allowing him to devote nearly all of his professional energies towards the elucidation of the factors outlined above. The investigator also has an excellent track record of mentoring and has trained several young people who are now fully independent scientists themselves. This award, therefore, would free up enough of the Principal Investigator's time to enable him to take on additional trainees in the future. The studies that are intended to address the above mentioned issues include human and animal experiments that are complimentary in nature. Well established rat models of B-vitamin depletion, aging, and colorectal carcinogenesis will be used to define which combinations of B-vitamin deficiencies and elder age can potentiate the procarcinogenic milieu produced by folate depletion, and by what mechanisms these effects are mediated. The identification of novel, previously unsuspected, pathways towards cancer will be elucidated by use of DNA microarray technology. A human study, designed to examine habitual cigarette smokers, will examine how smoking alters the metabolism of these vitamins in the mouth in order to convey an increased risk of oral cancer, and whether surrogate biomarkers of this cancer can be improved by the use of folate supplementation.

PI: Mazurana, Dyan
Title: Regional Gender and Generational Analysis of Armed Conflict, Peace and Justice Processes, and Disarmament, Demobilization, and Reintegration of Northern Uganda, Eastern Uganda, and Southern Uganda
Abstract:  The current research is a multi-year, regional, comparative study on the root causes and drivers of these armed conflicts. The study also examines the on-going peace and justice processes and the official and community-based disarmament, demobilization, and reintegration (DDR) programs and the shifts in livelihoods over the conflicts. The study is being conducted in Northern Uganda, Eastern Uganda, and Southern Sudan and will link up with current research underway by Tufts University in Darfur. The topics with the study are analyzed using a variety of perspectives to develop and enable a holistic understanding of the crises. In particular, we use gender and generational perspectives throughout the work, this includes paying attention to the roles, experiences, and voices of women and girls. In seeking more enduring solutions to peace and human security in countries and regions experiencing armed conflict, the United Nations Security Council Resolution 1325 and the United Nations Secretary-General Kofi Annan in his study Women, Peace and Security and subsequent Report called upon the Security Council and Member States to: Identify and utilize local sources of information on the impact of armed conflict, and the impact of interventions — peacekeeping, peace-building, humanitarian operations, disarmament, demobilization and reintegration programmes, and reconstruction — on women and girls, and on the roles and contributions of women and girls in conflict situations, including through the establishment of regular contacts with women’s groups and networks. These and other recent reports by the Secretary-General highlight the importance of research and gender analyses in developing better understanding of and responses to armed conflict, peace processes, and long-term peace building. Indeed, in the present study, the use of gender and generational perspectives has already revealed aspects of the conflicts that have not been adequately addressed in the past. In addition, these perspectives should make important contributions towards best policy and practice for the key processes underway in the region, most notably, peace negotiations, disarmament, demobilization, and reintegration, transitional justice, protection efforts, reconciliation, conflict prevention, and strengthening sustainable livelihoods.

PI: McKay, Diane
Title: The Effects of Walnuts on Antioxidant Capacity and Nutritional Status in Humans
Abstract: Epidemiological studies conducted in large populations have consistently shown that increased consumption of plant-based, antioxidant-rich foods, i.e., fruits, vegetables, grains, and nuts, reduces the risk for several chronic diseases. Among the various plants consumed worldwide, English walnuts (Juglans regia L.) have been reported to have the highest antioxidant activity, second only to rosehips (Rosa canina L.). The assay used to determine antioxidant activity in this study was the FRAP, which measures the reduction of Fe³⁺ to Fe²⁺ in the presence of antioxidants. There are a number of different compounds present in walnuts that are known to exhibit antioxidant activity, including 2 vitamin E (as γ-tocopherol), melatonin , and several non-flavonoid polyphenols (e.g., ellagic acid monomers and polymeric ellagitannins).

In vitro and ex vivo studies have demonstrated the ability of walnuts and walnut extracts to increase the resistance of low density lipoprotein (LDL) to oxidation. Walnuts and/or their constituents have also been shown to decrease levels of oxidative stress in diabetic mice and increase serum antioxidant capacity in rats. In humans, in vivo antioxidant capacity changes after consuming walnuts have only been measured in subjects with Type 2 diabetes. The main outcome of this human study by Tapsell et al. was not antioxidant capacity but rather changes in blood lipid profiles after consuming one of three different diets (one of which included walnuts). Furthermore, the Randox Total Antioxidant Status assay, which is used to measure the differences between these diets, is based on the ability of plasma to quench the synthetic radical cation 2,2’-azinobis (3-ethylbenzothiazoline-6-sulfonat), or ABTS, and capable of measuring antioxidant capacity in the hydrophilic compartment of plasma/serum only.

PI: McNinch, George
Title: Reductive Groups in Positive Characteristic
Abstract: The structure and representations of linear algebraic groups – and especially the reductive ones – are important to diverse parts of mathematics. The groups of their rational points over finite fields account for most of the finite simple groups; they are the natural transformation groups of many algebro-geometric questions; representations of the groups of their rational points over local and global fields carry deep number-theoretic information. The diversity of these features testifies to the mathematical significance – the intellectual merit – of the proposal.

The proposed work concerns reductive groups over – possibly imperfect – ground fields whose characteristic is a positive prime number p; this setting presents many important and interesting challenges.

The author recently gave a conceptual proof of a theorem of Testerman which locates a simple subgroup of rank 1 containing any given element of order p; the author’s construction used reduction modulo p techniques. Even more recently, he exploited a result in geometric invariant theory due to Kempf to obtain definitive conjugacy results for a class of optimal rank 1 subgroups. The reduction mod p techniques used earlier produce such optimal subgroups, and these subgroups now have a geometric characterization. For an element of order p which is rational over an arbitrary ground field, the techniques permit one to find a corresponding optimal subgroup of rank 1 subgroup defined over the ground field.

The proposed work will attempt to extend these results to characterize all rank 1 subgroups which are G-completely reducible in the sense of J-P. Serre. Moreover, it will seek the counterparts of these results for subgroups of rank greater than 1. It will also attempt to settle some of the mysteries associated with unipotent elements of order >p.

The author has also obtained recent interesting results on nilpotent orbits over ground fields. For example, he showed that the unipotent radical of the centralizer of a rational nilpotent element is split over the ground field; this result has important consequences for Galois cohomology. Over a local ground field, further applications include a proof that there are finitely many arithmetic nilpotent orbits, and a proof that nilpotent orbital integrals converge. This convergence was first proved by Deligne and Ranga Rao in characteristic 0.

The project suggests further related explorations. In particular, it seeks to combine the results just mentioned with the recent description of conjugacy classes in the component group of a nilpotent element in good characteristic, obtained jointly by Eric Sommers and the author, to obtain information about nilpotent orbits over, for example, the field of rational functions on an algebraic curve.

The broader impacts of the proposed research include the advancement of mathematical knowledge and learning, and the advancement of the mathematical working infrastructure through collaborations and the rapid electronic dissemination of the research results. The project will assist the graduate teaching role of the author. The project will support mathematical collaborations of the author with mathematicians at other institutions. Finally, the author will continue to make results of supported research available through use of the arXiv preprint server. This permits timely dissemination of the results, and supports a significant tool in the working infrastructure of modern mathematics.

PI: Mecsas, Joan
Title: Role of Yersinia Yops in an Animal Infection Model
Abstract: Our overall goal for this project is to determine how Yop virulence factors from pathogenic Yersinia pseudotuberculosis (Yptb) disarm host innate immune defenses during infection of mammals. Our hypothesis is that each Yop targets and inactivates different host defenses in different tissues and that distinct properties of each Yop are required in different tissues. YopE and YopO are two key Yops which are required for Yptb survival in the mesenteric lymph nodes (MLN) and GI tract of mice-two tissues which are chief targets of Yptb during infection. Our previous studies demonstrated that in the MLN, YopE targets oxidative burst generated by neutrophils while YopO targets oxidative burst generated by macrophages and potentially neutrophils. In contrast, YopE targets additional bactericidal factors in the GI tract because a yopE mutant does not survive in mice lacking the ability to generate an oxidative burst. YopE inactivates multiple small Rho-GTPases in cultured cells. Here we will investigate the contribution of each of its putative activities on colonization, of the MLN and GI tract to identify its targets during infection and we will investigate the basis for the different cell tropisms of YopE and YopO in the MLN. A secondary goal of this work is to test whether small molecules, which inhibit the translocation of Yops into culture cells, prevent a lethal infection by Yptb in mice. In summary, the work proposed here will provide insights into critical innate immune defenses targeted by YopE and YapO during infection of the GI tract and lymph nodes and advance our repertoire of therapeutics against pathogens with type III secretion systems.

PI: Merrigan, Kathleen
Title: Linking Agriculture, Food and Environment: An interdisciplinary Approach to Graduate Education
Abstract: The National Needs Graduate Fellowship application from Tufts University describes an interdisciplinary graduate program in Agriculture, Food and Environment from the School of Nutrition Science and Policy (SNSP). The AFE Program is unique, not only in its location with the only School of Nutrition in the US, but also by its scientific and applied training for Master's and Doctoral students interested in linking the environmental, socioeconomic and political implications and solutions for a sustainable global food system. The graduate training consists of:

1. Core curriculum, which includes courses designed to provide an integrated understanding of nutrition science, food and environmental policy
2. Research skills courses to equip students in designing and analyzing qualitative, quantitative and if needed, spatial data sets
3. Electives that represent a student's specialization (e.g. Watershed Management)
4. A 250-hour field internship
5. A directed study project

These directed study projects can be designed independently or pursued under one of the AFE Program research areas:

1. Green Consumer Markets and Food Labeling Standards
2. Strengthening the Scientific Foundation of Organic Livestock Production
3. Community and Local Food Systems
4. The Role of Small Farmers as Environmental Stewards
5. Connecting Sustainable Production Practices with Human Health

AFE students also participate in experiential learning opportunities through our partnership with the New Entry Sustainable Farming Project (NESFP) – a joint effort led by Tufts/AFE to assist immigrants with agricultural experience to apply skills in their new environment and become commercial farmers. Our student body has successfully grown over the last four years and our completion rates are high (near 100%). However, to meet our next challenge and expand our training capability, the AFE Program is looking to secure support by which we can attract more student diversity and thus, fresh perspectives for the future of agriculture and sustainability.

We hypothesize that "supplementation with E from an early age prevents or retards the development of atherosclerosis and the risk of CHD in individuals with either Western dietary habits (high fat, high cholesterol) or reduced intake of fat and cholesterol as suggested by American Heart Association." Since long-life E supplementation intervention studies in humans is rather costly and require a clinical trial of very long duration, this hypothesis needs to be tested first in an animal model. Thus, we propose to test this hypothesis in a double transgenic apo-B100xCETP mouse model [carries both human apolipoprotein B100 and human cholesteryl ester transferase protein (CETP) genes], and by inducing: a) moderate or b) high cholesterolemia from the age of 5wk and supplementing the diet of both groups with E starting from the ages of 5wk, 6mo and 12mo to the age of 18mo. We propose the following specific aims:

1. To investigate the interaction of E supplementation with moderate and high fat/cholesterol diets on the development of aortic atherosclerotic lesions when E supplementation is initiated at an early age compared to middle and later ages.

2. To determine the mechanism of E supplementation effect by studying the interaction of E with moderate and high fat/cholesterol diets on the nature and extent of atherosclcrotic lesions and presence of biological markers as evaluated microscopically, immunohistochemically, biochemically, and by molecular techniques including gene array technology.

These aims will be tested in apo-B100xCETP mice at the age of 18 mo following the dietary fat/cholesterol and E treatments using:

1. Light microscopy for en face determination of aortic lesions
2. Immunohistochemical techniques to detect and measure presence and extent of oxLDL, macrophages, smooth muscle cells, and T cell populations, as well as the expression of endothelial cell adhesion molecules, ICAM-1, VCAM-1 and P-CAM
3. Quantitative determination of rnRNA expression of aortic tissue for adhesion molecules. Inflammatory cytokines, and chemokines
4. Gene chip microarray techniques to determine responsive genes to dietary interventions
5. Analytical techniques for measurements of E and lipids in plasma and aorta

The molecular mechanisms of atherogenicity of high fat/cholesterol diet have beenextensively investigated. Furthermore, several studies have investigated the mechanism of vitamin E's effect. Most of the mechanistic studies and epidemiological observations support a beneficial role for E; however, the intervention studies have produced conflicting results. Thus the goal of this proposal is first to demonstrate the efficacy of E intervention from an early age in an appropriate model, while gaining insights into the mechanism of the effect using appropriate markers. A full and complete determination of the mechanisms will he proposed once the efficacy has been demonstrated.

The results generated from this study have several health implications. This study will provide information on the potential life-long enrichment of diet with E from an early age for the prevention of atherosclerosis and the reduction of the risk of CHD. It will also provide information on the effect of E supplementation combined with lowering fat and cholesterol intake from early life. It will also help us to understand the effectiveness of E supplementation initiated at a later age compared to earlier in the life as a preventive means to reduce the risk of CHD at a later age. The information generated from this study will be the basis for studying the impact of dietary E intervention and its interaction with dietary fat/cholesterol from childhood in the prevention of atherosclerosis in humans. The information from this study will also reveal the effectiveness of E supplementation initiated in middle age on the reduction of the risk of CHD.

PI: Meydani, Simin
Title: Aging, Vitamin E and Immune Function in Aged
Abstract: Aging is associated with a decline in T cell mediated functions, which contributes to a higher incidence of, and morbidity and mortality from, infectious diseases and tumors. The age-related defect in T cells has been shown to be due to intrinsic declines in T cell function, as well as increased production of prostaglandin (PG) E2, a T cell suppressive factor, by macrophages (Mphi). We demonstrated that the increased PGE2 production was due to ceramide mediated upregulation of cyclooxygenase 2 (COX-2) transcription, a key regulatory enzyme in PGE2 synthesis. The signaling mechanism through which ceramide upregulates COX 2 expression, however, is not known and needs to be investigated. We further showed that vitamin E (E) supplementation improves T cell mediated function by two distinct mechanisms:

• By decreasing PGE2 production, thus reducing macrophage Mphi mediated suppression
• By directly enhancing T cell function, independent of its effect on Mphi PGE2 production

E exerts its effect by improving the ability of naive T cells from old mice to produce IL-2 and progress through cell division cycles. The mechanism of E-induced enhancement of naive T cell function is not known and needs to be determined. Thus, the specific aims of this proposal are:

1. To determine the signaling pathway involved in ceramide-induced upregulation of COX-2 expression in old Mphi. To accomplish this goal, we will test the hypothesis that ceramide upregulates COX-2 expression in old macrophages through enhancing PKC-zeta activity, leading to increased IkappaB phosphorylation, and thus degradation. This in turn will increase NFkappaB activation and COX-2 expression.

2. To determine the mechanism of E-induced increase in the function of old naive T cells. To accomplish this goal, we will test the hypothesis that E enhances naive T cell function in old mice by increasing their ability to form an effective immune synapse at the site of T cell receptor (TCR) and antigen contact. This, in turn, will lead to improved TCR-associated signal transduction, and subsequent, IL-2 production in old mice. We propose that E induces its effect by changing the redistribution of key TCR associated signaling molecules in membrane lipid domains, known as lipid rafts, by one or both of the following mechanisms:

   1. Increasing palmitoylation of key signaling molecules associated with TCR-mediated activation
   2. Changing the structure of the lipid component of lipid rafts.

These experiments will elucidate the mechanism of the age-related dysregulation of macrophages and T cells, as well as their normalization by E. This, in turn, will help in designing practical nutritional interventions to reverse and/or delay the age-associated dysregulation of immune and inflammatory responses as well as diseases associated with it.

PI: Miczek, Klaus
Title: Behavioral Neurobiology of Aggression: Alcohol, Gaba, and 5-HT
Abstract: Epidemiological and criminal statistics as well as neurobiology, and pharmacotherapy all provide converging evidence that suggests links between alcohol consumption, impulsivity, and pathological aggression. The proposed research aims to dissect these links at the behavioral and neurochemical level, with particular focus on the relative role and interactions between GABAA and serotonin systems. A first specific aim seeks to analyze how alcohol-heightened aggression is related to other forms of escalated aggression. Experiments are designed to test the hypothesis whether or not a common behavioral profile of varied forms of escalated aggression characterizes individuals who engage in alcohol-heightened aggression. The second and third aims focus on pharmacological tools that are employed to characterize the relative contribution of 5-HT1A, 5-HT1B, and GABAA receptors, their pre- versus post-synaptic sites in brainstem, and prefrontal cortical regions in animals that engage in alcohol-heightened aggression. The fourth aim examines how serotonergic modulation of GABAergic systems determines alcohol-heightened aggression. Inversely, how do neuropharmacological manipulations of the modulatory sites on the GABAA receptor, particularly via neurosteroids, enable 5-HT-mediated effects on aggressive behavior? Pharmacological experiments are designed to stimulate pre-synaptically 5-HT1A and 5-HT1B receptors or to neurotoxically lesion raphe nuclei in order to assess the importance of serotonergic inhibition on the activating effects of positive modulators like neurosteroids on alcohol-heightened aggression. A fifth aim is directed at the neural sites of the GABAergic and serotonergic mechanisms that are critical for the aggression-heightening effects of alcohol. Intracranial microinjections are used to determine whether activation of 5-HT receptors in the raphe nucleus or in terminal forebrain regions are the critical sites for reducing escalated fighting. Conversely, can blockade of the 5-HT autoreceptors in the raphe n. potentiate the aggression-heightening effects of alcohol and other positive modulators at GABAA receptors?

PI: Miczek, Klaus
Title: Ethanol-Heightened and Escalated Aggression in Mice
Abstract: Alcohol-induced aggression is a serious problem with serious, far-reaching consequences for individual and public health. The proposed study is expected to contribute new evidence to the existing body of knowledge of ethanol-heightened and other forms of escalated aggression. The project will examine individual differences in ethanol self-administration, vulnerability to ethanol-induced aggression, aggression following social-instigation, and escalated aggression following extinction of scheduled reinforcement. The research will utilize an animal model (i.e. mice), well-established ethanol self-administration techniques, the resident intruder paradigm, and detailed behavioral analyses. Escalated aggression may be due to interactions of GABA and 5-HT in mesocorticolimbic neurons. Therefore, the present project will investigate the role played select neurotransmitter systems, specifically GABA and 5-HT, in escalated aggression employing pharmaceutical receptor manipulations to determine if neurotransmitter systems play a predictive role in [modulating such behaviors. Thus, the present research should enhance current understanding of individual characteristics and neurochemical mechanisms involved in alcohol-heightened aggression.

PI: Miczek, Klaus
Title: Psychomotor Stimulants and Aggressive Behavior in Animals
Abstract: Epidemiological data, statistics on perpetrators of violent crimes and of victims of violence under the influence of drug as well as neurobiological evidence link social stress and drug use. The proposed research aims to characterize the common behavioral and neural features of individuals who experience salient types of social stress and who self-administer psychomotor stimulants and opioids in a compulsive-like manner, and to define the neural circuits for these apparently opposing activities.

Ostensibly aversive events such as social defeat stress and euphorogenic effects such as those produced by cocaine or opioids share physiological and corticolimbic circuits. First, experiments are designed to identify the critical behavioral and mesocorticolimbic features of brief episodes of social defeat stress, chronic subordination stress, and impulsive aggressive behavior as potential determinants for the transition to compulsive-like intravenous self-administration of cocaine, d-amphetamine or morphine. We aim to characterize the transition from regulated to dysregulated drug taking by studying the long (>24 h) drug binge as a model.

Secondly, the neurobiological mechanisms for the behavioral sensitization that results from repeated episodes of social defeat stress will be characterized with a focus on glutamatergic, GABAergic and serotonergic modulation of mesocorticolimbic dopaminergic pathway. In vivo microdialysis measurements aim to reveal the emerging neural sensitization in the microcircuit consisting of amgydaloid and cortical structures modulating the VTA DA system.

Thirdly, neuropharmacological experiments are designed to examine the role of NMDA and AMPA glutamatergic receptos, GABAA receptors, and subtypes of the 5-HT receptor families in modulating activity in the mesocorticolimbic DA pathway for their relevance in sensitization to social stress and in the transition to intense dysregulated cocaine and morphine self-administration in stress-sensitized animals.

It is anticipated that the sensitizing effects of social stress experiences can be reversed or protected against by targeting the modulatory influences on the VTA system. Reversal of cross-sensitization from stress may be a means by which to prevent the transition to out-of-control compulsive-like drug taking.

PI: Milbury, Paul
Title: Bioavailability and Metabolism of Anthocyanins Following Acute Cranberry (Vaccinium Macrocarpon) Consumption and Their Correlation with Vascular Responsiveness
Abstract: Little is known about the acute or chronic pharmacokinetics and metabolism of anthocyanins consumed in amounts relevant to usual dietary intakes. Studies of pharmacologic doses of anthocyanins reveal a greater than anticipated level of methylated metabolites that may be a function of high doses. No previous study has directly investigated the correlation between cranberry anthocyanins and relevant physiological responses, including vascular reactivity. Thus, we hypothesize that anthocyanins from a single oral dose of cranberry juice will be absorbed quickly and can be measured in the circulation and urine within 2-4 h. Further, these anthocyanins will undergo a degree of metabolism but will circulate as well as be removed from blood as both parent compounds and metabolic products. The presence and concentration of these total and individual compounds will be correlated with vascular responsiveness determined by brachial artery reactivity measured during the protocol.

PI: Minear, Larry
Title: The Humanitarian Agenda 2015: Principles, Power, and Perceptions
Abstract: Starting in January 2006, an international and interdisciplinary team will investigate these issues in Afghanistan, Colombia, Iraq, Liberia, Burundi, and the Sudan. Additional countries may be added if finances permit. Members of the team are Sippi Azarbaijani-Moghaddam, Antonio Donini, Greg Hansen, Larry Minear, Tasneem Mowjee, Karma Purushotma, Ian Smillie, Elizabeth Stites, and Xavier Zeebroek.

A detailed methodology for data collection and survey tools has been developed. In each country, the researchers will focus on a key theme (e.g. terrorism and counter-terrorism in Colombia; perceptions of aid agencies in Afghanistan; integrated missions in Liberia and Burundi; etc.) while, at the same time, collecting information on the other three themes. This should allow a composite picture to emerge of the challenges and opportunities facing humanitarian agencies in countries in crisis.

This picture will be built from the bottom up. The emphasis of the data collection is on the perceptions of local communities and local actors. Working with the support of local partner agencies, researchers will conduct individual and focus group interviews with members of local communities as well as aid agency personnel in the field. This will be complemented by interviews with a wide range of informants in the donor, UN, NGO and academic communities as well as by a survey of some 1000 aid personnel at donor and agency headquarters using a web-based electronic questionnaire.

The team will come together for a week in May to discuss findings and agree on the main recommendations of the research. The team’s findings will be the subject of a number of reports and articles scheduled for completion in the latter half of 2006. These will include a final report and several technical papers or case studies as well as shorter policy papers aimed at aid agencies and donor institutions.

PI: Minear, Larry
Title: The Humanitarian Agenda 2015: Politics, Power, and Perceptions
Abstract: Conventional wisdom has it that the last decade has seen a significant deterioration in the working environment for humanitarian agencies and a corresponding increase in attacks against aid workers. High-profile attacks and targeted killings in Afghanistan, Iraq, Chechnya, Sudan, the DRC, and elsewhere have reinforced the impression that the humanitarian profession is becoming more dangerous and more reviled by militant groups and others who choose to deliberately attack aid workers. 

This may well be the case, but there is little hard data to back up this impression. Are there more attacks on and killings of aid workers than, say, 15 years ago — or is it that we pay more attention to such incidents today? Are there more attacks relative to the numbers of aid workers or fewer? In other words, are there more attacks because there are more aid workers active in war zones or are we simply recording them more accurately?

While a few studies have attempted to analyze data on such incidents, the data itself is patchy, limited mostly to incidents involving international staff, and does not lend itself to disaggregating the motives behind the attacks. When an incident occurs — for example, an aid worker is shot at on her way to work — we often do not know if the attack was politically-motivated, a random episode of opportunistic petty criminality, the result of a personal dispute, a sexually motivated aggression, or a case of mistaken identity. Was she attacked because of who she is, what she represents, what she does, what she is perceived as doing, because she happened to be there, or because she was involved in a local dispute of a personal, professional or commercial nature? 

While statistically speaking the overall occurrence of incidents involving aid workers remains shrouded in fog, it is nevertheless possible to try to identify some qualitative trends, especially at the country level. For example, in Afghanistan before 9/11 and in Iraq before the US-led intervention, there had been very few attacks against international aid agency staff. Today this is no longer the case. For reasons that would be too simplistic to connect exclusively to the “world re-ordering” that has taken place in these two countries, it is now no longer taboo to attack aid workers, local or international, and the price paid by the aid community has been dramatically high.

Mechanisms of social protection and control — the contract of acceptability that permitted aid workers to work in relative security — have been deeply affected, or so it would seem. Whether the attacks are politically motivated, criminal, opportunistic, or of a personal nature, the worrying fact is that they are taking place at all in places where aid workers were relatively secure only a few years ago. The objective of the research is to gain a more detailed and sophisticated understanding of the political economy of attacks against aid workers in insecure settings.  Specifically, through analysis of existing data and field-based research using questionnaires and focus group interviews in 3-4 countries, the research will seek to: 

1. Develop a typology of the motives/rationales of attacks against aid workers in conflict settings

2. Analyze available data and also data collected through the proposed research to provide a qualitative and quantitative picture of the motives of attacks against aid workers

3. Produce and disseminate a number of analytical research outputs contextualizing staff security issues within the wider ambit of the physical and human security of local communities

PI: Moore, Claire
Title: The Coupling of mRNA Transcription and 3' End Formation
Abstract: The emerging model of eukaryotic mRNA synthesis is that transcription and mRNA processing events are carefully orchestrated in vivo by a physical association of the different machineries. For example, RNA polymerase II affects the efficiency of 3' end processing, and processing factors affect the efficiency of transcription termination downstream of poly(A) sites. We are interested in the precise molecular mechanisms involved in the coordination of these two events and have identified several new points of interaction between transcription and cleavage/polyadenylation factors. These findings suggest that the presence of processing factors at the promoter might affect the efficiency and/or specificity of transcription initiation and facilitate recycling of RNAP II back to the promoter for another round of transcription. This may serve as a mechanism to insure the proper loading of processing factors onto the transcriptional complex, and in turn, the subsequent polyadenylation of the transcript, which is essential for optimal export, translation, and turnover of mRNA. To investigate this issue, we propose the following specific aims:

1. Can the activity of Ssu72 in transcription be separated from its role in 3" end cleavage? We have found that Ssu72, previously identified as a protein affecting initiation, is directly involved in mRNA 3'end cleavage. We will analyze an existing collection of ssu72 mutants to try to separate the cleavage activity of Ssu72 from a function in transcription initiation and develop new assays to help discriminate these functions.

2. What is the functional significance of the interactions of Sub1 and Ssu72 with Pta1, and how are these interactions regulated? Ssu72 and Sub1 were initially identified based on genetic interactions with TFIIB. We have found that these proteins genetically interact with the Ptal subunit of Cleavage/Polyadenylation Factor (CPF). Moreover, they physically bind Pta1 in a mutually exclusive manner. We will test the hypothesis that sequential interactions of Pta1 with Ssu72 and Sub1 are important for efficient initiation and/or cleavage of pre-mRNA.

3. Does Swd2 function in mRNA synthesis as part of CPF? This protein is intimately associated with CPF and the Set1 histone methylase. However, Swd2 depletion has no effect on 3' end processing, but causes inefficient transcription termination and reduced mRNA levels.

We will test the hypothesis that Swd2 affects termination by recruiting Set1 to the transcription complex. We will identify the contact point of Swd2 with CPF and examine how disruption of this interaction affects mRNA synthesis. A genetic screen will be used to identify other important functional interactions with Swd2.

PI: Moore, Claire
Title: Bringing Engineers into New Disciplines
Abstract: In this application we propose to create a Short Term Training Institute for undergraduate students in science and engineering to train in several aspects of biomedical research centered around the theme of type 2 diabetes. The goal is to bring engineers and scientists together in multi-disciplinary teams to solve new problems in health-related biomedical research that have not been traditional avenues for bioengineering. The program will be called BEND (Bringing Engineers into New Disciplines), and will involve ten undergraduates for ten weeks during the summer. The students will have didactic and conference courses designed to give them new understanding of biomedical and behavioral problems related to treatment of diabetes. They will also work in small teams with an engineering graduate student mentor and two faculty mentors, one from biomedicine and the other from engineering, to investigate solutions to problems identified by the program faculty through round-table discussions that have occurred during the previous year. The students will present their results at a symposium at the end of the program, and possibly continue to work on the project as a senior honors thesis. Tufts University has two assets that will be combined to create this program. The first is a history of successful undergraduate research training in several individual disciplines, including engineering and biomedicine. The second is a history of and a developing strength in interdisciplinary research. Tufts is poised to create a program that will not only give students in the quantitative sciences direct experience in clinically relevant biomedical research, but also brings together a diverse group of faculty for new avenues of research cooperation.

PI: Moore, Claire
Title: Biomedical Research Experiences for Engineering Majors
Abstract: Tufts University seeks NIGMS funding for a Summer Internship Program designed to provide biomedical research experiences to Tufts undergraduate students majoring in engineering and computer science. The long-range goal is to encourage cross-disciplinary training for the next generation of biomedical scientists and thus promote an interdisciplinary approach to solving problems related to human health. The specific goal is to increase the number of students who pursue careers in biomedical research. The objectives are:

1. To increase the understanding of students about what a career in biomedical research would entail through distinct, innovative summer research internships on the Tufts Health Sciences Campus

2. To increase the students' awareness of the benefits of biomedical research and potential careers, collaborations and post-baccalaureate training opportunities through workshops and seminars

3. To increase students' laboratory skills and confidence by their planning, completing, and presenting a mentored, independent hands-on biomedical research project

4. To increase the number of students who choose Biomedical Engineering as a major or minor

5. To increase the number of graduating engineering seniors who pursue post-baccalaureate training in bioengineering or biomedical research or enter industry careers in these areas

This program consists of a ten-week summer research internship for ten students in which the students interact with and work alongside outstanding research scientists. It builds upon existing strengths at Tufts in engineering, biomedical research, and undergraduate education, a proven commitment of Tufts to undergraduate research as a teaching tool, and a thriving culture of inter-departmental and inter-school collaborations.

By exposing the students to potential opportunities and benefits of collaborations with biomedical researchers, it will help the students decide if biomedical research is an area in which they would like to apply their engineering and computer science training.

PI: Moore, Claire
Title: Training in Education and Critical Research Skills
Abstract: This is a resubmission of our application to establish the Training in Education and Critical Research Skills (TEACRS) Program at Tufts University in response to the Institutional Research and Academic Career Development Award Program. Our program is based on the need for university faculty that are prepared to meet the multiple challenges faced by young Assistant Professors pursuing their first independent position. We will provide our postdoctoral trainees with the career skills they will need to succeed in an academic research environment by integrating three training components: rigorous bench research; mentored teaching experience; and career development skills. By partnering with two local minority serving institutions, Roxbury Community College and the University of Massachusetts, Boston, our trainees will also acquire a greater knowledge and sensitivity to diversity issues and provide enriched course material to institutions serving a minority population. Our faculty of 42 is committed to research excellence and has a strong training record. Research opportunities focused on multiple biomedical problems with special emphasis on basic biological problems, cancer, neuroscience and infectious disease will be available to trainees. Research committees and research advisors will monitor progress and career development with respect to research. The teaching component uses workshops, discussion groups and tutorials to prepare trainees to develop and present a course of their own design at one of the minority serving institutions. A series of career building workshops, along with checkpoints that provide mentoring and constructive feedback round out the program. Our goal is to produce the academic scientific leaders of the future. University faculty must develop independent research programs, an activity that requires creative thought, high intellectual capacity, outstanding facility with oral and written communication and considerable experimental skill. These activities must be sustained while faculty participate in classroom teaching, serve on a variety of committees and mentor trainees. Success also requires additional skills in interpersonal relations, budgeting and time management. The TEACRS program will provide trainees with the research portfolio needed to compete for academic positions and the skill set that postdoctoral fellows need to meet the challenges they will face as they enter academic careers.

PI: Morgan, John
Title: Fernald Dental Program
Abstract: Under this contract, Tufts Dental Facilities will provide comprehensive dental services to the residents of Massachusetts’s developmental centers serving people who are mentally retarded and to individuals with developmental disabilities who reside in the community. Priority will be given to mentally retarded who are class clients. This care includes all basic dental services such as preventive, restorative, periodontal, endodontic and oral surgery, with a focus on prevention and education.

In addition, the program will train dental residents so that after graduation they are available to care for Massachusetts’s citizens with developmental disabilities and other people served by human service agencies.

PI: Morgan, John
Title: Oral Health Access Across the Commonwealth
Abstract: The project, Oral Health Across the Commonwealth (OHAC), is a statewide initiative to provide preventive oral health services to early childhood students, Head Start students, and children with special health care needs using portable equipment at the school sites.

The project has four interrelated components that include:

1. Oral health education
2. Oral health assessment
3. Oral health prevention including prophylaxis, fluoride varnish and oral hygiene instruction
4. Referral and case management services

The educational component includes in-service training for school nurses, teachers, and other related staff members to review basic oral health knowledge and to insure proper commitment to the project. The oral health curriculum, called “Open Wide”, and developed by the Connecticut State Department of Public Health, is used as a training tool for this purpose.

The participating students receive group presentations on "What to Expect at the First Dental Visit" using curricula developed by the Arizona Department of Health Services. In addition, students receive individualized oral hygiene instruction and a toothbrush.

PI: Morgan, John
Title: Support for Dental Services for DMC Clients FY2007
Abstract: The Bureau of Family and Community Health of the Massachusetts Department of Public Health (MDPH) is soliciting proposals from qualified bidders for a program which provides comprehensive dental services to the residents of Massachusetts Developmental Centers (DC) serving people who are mentally retarded and individuals with developmental disabilities who reside in the community. Priority is given to mentally retarded who are class clients.

PI: Naumova, Elena
Title: Gastroenteritis and Extreme Weather Events in Elderly
Abstract: 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)

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.

PI: Navia, Bradford
Title: In Vivo Proton MRS Studies of Cerebral Injury in HIV Infection
Abstract: Highly active antiretroviral therapy (HAART) has resulted in a marked rise in survival rates among HIV-infected persons. To what extent the brain compartment benefits from such treatment, particularly in the setting of advanced, even stable disease, remains unclear. Recent studies suggest that cerebral injury and neurocognitive impairment can persist or unfold in the setting of HAART and stable disease. The frequency, spectrum and extent of neurological injury, its relationship to cognitive functioning and responsiveness to antiretroviral therapies thus remain important yet unresolved issues.

The HIV MRS consortium has identified regional and cellular abnormalities specific to different stages of HIV-associated cognitive impairment (AIDS dementia complex, ADC) as well as an in vivo model of brain injury that suggests basal ganglia and neuronal events may be critical to the development of ADC. It has also identified potential host (e.g. age, plasma MCP-1, CSFIP-10, CSF fractalkine) and viral factors (CSF HIV RNA) at baseline that may increase the risk for neurocognitive impairment (NCI) or its further decline.

This is a five-year multicenter longitudinal study of cerebral injury (as measured by MRS) and NCI in 310 HIV-positive subjects with a history of advanced immuno-suppression (nadir CD4<100/mL) on HAART. The study will fill an important gap in the literature by implementing MRS in a prospective design in order to identify subjects at risk for NCI. We will examine the pattern and dynamics of regional injury by MRS among 250 neurologically asymptomatic (NA) and 60 ADC subjects.

The majority of subjects will be enrolled from three well characterized existing prospective cohorts, including two ACTG studies (N= 210) and from the brain bank studies UCLA NNAB and UCSD CNTN (N= 100). Subjects will be stratified by CD4 count at entry, <200 or >200 cells/ml. The brain bank subjects will be further divided into virologically suppressed and nonsuppressed groups.

The primary aims are:

1. To identify NA subjects at risk for brain injury as measured by MRS and examine host (e.g. age, chemokines) and viral factors (HIV RNA) at baseline that may increase the risk for subsequent neurological injury and NCI

2. To determine whether the emergence of basal ganglia and neuronal factors predict the onset of NCI or its progression

3. To examine dynamic relationships between neurologic function, biomarkers that reflect changing virologic and immunologic activity, and changes in treatment

Multivariate longitudinal models and variable selection will be used to study the neurological effects of HIV infection as a dynamic system of interrelated factors. Detection of brain injury among NA subjects on HAART and at risk for ADC will have critical ramifications for overall health outcome and early therapeutic intervention.

PI: Nelson, Miriam
Title: Promoting Heart Health in Midlife and Older Women
Abstract: Heart disease is still considered a man's problem, despite the fact that every year since 1984, it has affected more women than men in the U.S. A woman dies from heart disease about once every minute, for an annual total of about half a million. That makes it the leading cause of death for women in this country, taking six times as many lives as breast cancer. In addition, it is the leading cause of morbidity in women, resulting in eight million women living with heart disease and its adverse consequences.

Still, most women go unprotected from heart disease, unaware of the risk, undiagnosed once they have it, and insufficiently treated once they're diagnosed relative to men. New data presented at the American Heart Association meetings in November 2005 confirmed that since the mid-1990's the situation has not improved, even after the issue of heart disease has been thrust into public consciousness. Making matters worse, women and their physicians tend to downplay any red flags they might experience because they aren't expected to have heart problems.

Despite these grim statistics, research shows that following a lifestyle that includes a healthy diet, weight control, and appropriate physical activity can dramatically reduce the risk of heart disease in women. A dietary pattern that focuses on vegetables, fruits, low and nonfat dairy foods, whole grains, legumes, fish, and lean proteins helps to reduce cholesterol levels, lower blood pressure, and improve weight control, which lead to an overall reduction in heart disease risk. Increasing physical activity similarly helps to improve weight control and reduce risk of developing heart disease in women.

However, the public health community has fallen short in disseminating and translating this critical information, particularly to midlife and older women and their clinicians. For instance, few women realize that their risk of developing heart disease increases dramatically after menopause. Even if they are aware of it, the solution that many middle aged and older women relied on, hormone replacement therapy has been shown ineffective and may actually increase risk. It is clearly time to raise awareness and provide women with strategies and tools that help them adhere to a heart healthy diet and regular physical activity. An added benefit is that they can serve as role models for their daughters, granddaughters, and friends.

Strategic prevention tactics involve the development and evaluation of educational and behavioral programs that can be implemented by community organizations where many higher risk women can be reached. Currently, there are smoking cessation programs in almost every community in the country. In contrast, there are very few (if any) nutrition and physical activity programs targeted to reduce heart disease in midlife and older women.

This project proposes to use a community coalition between the Strong Women Program and the Cooperative State Research, Education, and Extension Services (CSREES) of the United States Department of Agriculture (USDA) in Arkansas and Kansas as pilot programs to develop, implement, and evaluate The Healthy Heart Program for Midlife and Older Women. If effective, the program can be replicated across the country, with minor modifications to adapt to regional differences in eating patterns and available venues for physical activity.

PI: Nurminskaya, Maria
Title: Evaluation of T-Gases as Regulators of VSMC
Abstract: Cardiovascular calcification (mineralization) is a common pathologic condition associated with ageing, diabetes, and chronic renal insufficiency. It often leads to stroke, amputation, and ischemic heart disease. The contribution of vascular smooth muscle cells (VSMCs), trans-differentiated to acquire the osteoblast phenotype, in depositing mineralized matrix is well established. However, the mechanisms and regulators of such trans-differentiation are not understood yet.

We hypothesize that extracellular enzymes transglutaminases (T-Gases) may induce VSMCs to deposit mineralized matrix. This hypothesis derives from our published and preliminary data including those, obtained from the studies currently funded by AHAF:

1. Co-localization of T-Gases with areas of calcification in the arteries
2. Induction matrix calcification in VSMC cultures by exogenous TGase (TG2 or tissue transglutaminase)
3. The ability of TG2 to enhance expression of osteopontin, one of the osteoblast-specific proteins, in VSMC
4. Binding of TG2 to LRP receptors on VSMC

To further extend our analyses of the role of TGases in the regulation of vascular calcification, the following specific aims are proposed:

1. To examine the ability of another TGase - FXIIIa, which is often accumulated in the areas of pathologic arterial calcification in human patients - to induce matrix mineralization and acquisition of osteoblast phenotype by VSMC in vitro. Osteoblast maturation will be assessed by RT-PCR and Osteogenesis Arrays and by histological analysis of matrix mineralization.

2. To determine the role of TG2/LRP binding in the mediating the TG2-induced osteopontin expression and matrix mineralization in mouse VSMC cultures. For this, we will express and purify the receptor-associated protein (RAP), which non-specifically inhibits LRP-mediated signaling. We will analyze the TGase-induced gain of osteoblastic phenotype in VSMC with altered LRP-mediated signaling, by assessing the expression levels of osteopontin and the levels of matrix mineralization.

Identification of intracellular pathways mediating the TGase-induced matrix mineralization may be of potential interest in developing novel therapeutics for vascular calcification.

PI: Obin, Martin
Title: Adipocyte Necrosis and Adipose Tissue Inflammation: A New Model of Macrophage Recruitment and Activiation in Obesity
Abstract: Inflammation of white adipose tissue (WAT) is a critical event in the pathogenesis of obesity disorders, including the metabolic syndrome and type 2 diabetes. Obesity-associated WAT inflammation, and the development of systemic insulin resistance are due in large part to recruitment of bone-marrow derived macrophages to WAT and the subsequent release of proinflammatory mediators by these recruited macrophages. However, neither the precipitating event(s) nor function(s) of macrophage infiltration into adipose tissue are known.

This proposal addresses this fundamental gap in our understanding of WAT inflammation. The proposal is based on preliminary studies that reveal, for the first time, that:

1. The preponderance of macrophages in WAT are localized almost exclusively to necrotic adipocytes.
2. The frequency of adipocyte necrosis increases dramatically in obesity.
3. At sites of adipocyte necrosis, macrophages fuse to form syncitium that surround, sequester and scavenge adipocyte debris, in particular, the residual 'free' lipid droplets.
4. The 'free' lipid droplets of necrotic adipocytes act as foci of chronic macrophage activation, inducing the formation by macrophage syncitia of multinucleate giant cells (MGCs).

Based on these novel observations we hypothesize that adipocyte necrosis is an underlying event linking increased adiposity with macrophage localization, activation and persistence in adipose tissue and with the development of adipose tissue inflammation and insulin resistance.

PI: Ordovas, Jose
Title: Gene/Diet Effects on Plasma Lipoprotein Levels
Abstract: Cardiovascular diseases (CVD) are the major causes of morbidity and mortality in the US and their risks are determined by non-modifiable (i.e., genetic) and modifiable factors (i.e., plasma lipid levels). It is well established that low levels of high-density lipoprotein cholesterol (HDL-C) are associated with increased CVD risk. A key determinant of HDL metabolism is hepatic lipase (HL), an enzyme that catalyses the hydrolysis of triglycerides and phospholipids in lipoproteins. Within the hepatic lipase (LIPC) gene at position -514 exists a common C/T polymorphism. The -514TT genotype is associated with a 40% decrease in HL activity and increased HDL-C levels. However, this effect is variable among populations, suggesting gene- environment interactions. One of this grant's earlier key findings is that the -514 polymorphism shows a statistically significant interaction between HDL-C levels and dietary fat intake in Whites. Moreover, we replicated the findings in an Asian population. What remains to be demonstrated is whether these interactions occur when subjects modify their dietary fat intake and if so, what mechanisms are involved. Thus, the primary goal of this proposal is to use an interventional setting to thoroughly evaluate interactions between LIPC gene variants, changes in dietary fat intake and plasma lipid levels, inflammation markers and endothelial function. Moreover, we hypothesize that minorities, such as Hispanics, due to the enrichment of the -514 T allele (-0.36) are more susceptible to the effects of these gene-diet interactions. Hence, we propose a randomized, cross over, two-phase diet intervention aimed at assessing the impact of consuming different levels of dietary fat (15% and 37%, respectively, 6 weeks each) on plasma HDL related measures in Caribbean Hispanic subjects (n=80; 40 CC and 40 TT). Our main hypothesis is that CC subjects respond to increases in the % of energy consumed as fat with increases in HDL-C levels. Conversely, in TT subjects, such dietary change results in decreases in HDL-C levels and worsening of endothelial function. Therefore, T subjects will have an elevated CVD risk when consuming high fat diets. Moreover, given the high frequency of the T allele in Hispanics, this interaction could account for some of the increase in CVD risk seen after acculturation. This evidence-based knowledge will contribute to a rational approach toward more effective dietary recommendations aimed to decrease health disparities and to a better CVD prevention.

PI: Ortiz, Daniel
Title: ABC-Transporter Binding Proteins and Trafficking
Abstract: The ABC (ATP-Binding-Cassette)-type protein BSEP (ABCB11) is essential for bile formation. BSEP resides primarily in the canalicular membrane of hepatocytes and mediates ATP-dependent transport of conjugated bile acids from cytoplasm to bile. Correct targeting and recycling to the apical membrane are fundamental to BSEP function and regulation. Mutations that cause BSEP retention in the endoplasmic reticulum produce Progressive Familial Intrahepatic Cholestasis type II. Additionally, BSEP removal from the canalicular membrane is associated with drug and sepsis induced intrahepatic cholestasis. BSEP mobilization and targeting to the apical membrane are mediated by association with factors that connect the transporter to sorting and trafficking networks.

We previously identified proteins that interact with BSEP in the apical domain and regulate its plasma membrane delivery and endocytosis. However, there is no information regarding proteins that bind BSEP and regulate its progress through the early secretory pathway or exit from the Trans Golgi Network, which represents the central sorting hub of the vesicular transport network. We have identified two proteins, a clathrin adaptor and a putative endoplasmic reticulum sorting receptor, which interact with BSEP and influence its apical membrane expression. The goal of the proposed research is to investigate the role these proteins play in regulating BSEP trafficking.

Experiments in Aim 1 will clarify which aspect of the BSEP trafficking itinerary in polarized cells and hepatocytes is controlled by interaction with the clathrin adaptor.

Aim 2 focuses on determining the extent to which accessory proteins, which form a macromolecular complex with BSEP and the clathrin adaptor, participate in BSEP delivery to the plasma membrane.

Specific Aim 3 will investigate the relevance of an endoplasmic reticulum resident protein, which directly interacts with BSEP, to transporter maturation and vesicular trafficking to the Golgi. We will also ascertain whether overexpression of the interacting protein improves trafficking of BSEP PFIC II mutants.

Inherited or acquired defects that compromise BSEP expression in the canalicular membrane are likely causes of intrahepatic cholestasis. Therefore, elucidating pathways that govern transfer and recruitment of ABC-transporters to the apical membrane, and identification of proteins which control these processes, will provide critical insight into mechanisms underlying cholestasis and suggest targets for drug design.

PI: Panjwani, Noorjahan
Title: Corneal Epithelial Cell Surface Glycoconjugates
Abstract: The failure of the epithelium to migrate over the wound or of the migrated epithelium to remain adherent to the substratum may lead to the development of a number of debilitating clinical conditions of the cornea including recurrent erosions and persistent epithelial defects. We established during the previous funding period that two carbohydrate binding proteins, galectins-3 and -7, are among the key molecules which mediate corneal epithelial cell migration.

For an understanding of the mechanism by which galectins-3 and -7 mediate corneal epithelial cell migration, in Aim 1, we shall identify and characterize the corneal epithelial cell surface and extracellular matrix (ECM) glycoproteins which serve as counterreceptors of galectins-3 and -7, and will establish whether the lectins modulate corneal epithelial cell migration by binding to well-known integrins, growth factor receptors, and/or ECM molecules.

In Aim 2, using small interfering RNA (siRNA) and/or antisense adenoviral constructs, cDNA microarrays and glycogene arrays, we shall determine whether galectin-3 mediates corneal epithelial cell migration indirectly by modulating the expression of key adhesion and/or signal transduction molecules.

In Aim 3, by determining whether galectins-3 and/or -7 modulate the activation of specific kinases (focal adhesion kinase, protein kinase B, MAP kinases) that are well known for their role in cell migration, we shall establish whether the lectins mediate corneal epithelial cell migration by modulating specific signal transduction pathways.

The proposed studies will contribute to a better understanding of the molecular basis of corneal epithelial cell migration and should ultimately help find novel therapeutic strategies for treating nonhealing corneal wounds.

In addition, this study will contribute to the basic understanding of the general disorders of impaired or delayed re-epithelialization including chronic wounds in the elderly, decubitus ulcers, and venous stasis ulcer of the skin, conditions that together affect millions of individuals worldwide.

PI: Panjwani, Noorjahan
Title: The Role of Selectin-Mediated Recognition in Glaucoma
Abstract: Glaucoma is the second leading cause of blindness in the world after cataract, affecting approximately 70 million people. Elevated intraocular pressure due to the obstruction of the aqueous outflow pathway is a major causal risk factor in the development of primary open-angle glaucoma. Treatment of this blinding disease is compromised because of the lack of understanding of the molecular mechanism responsible for the resistance to the outflow facility. In a recent study, a carbohydrate-binding protein, E-selectin (ELAM-1, CD62E), was identified as a specific molecular marker of glaucoma. It was demonstrated that while E-selectin is consistently present on trabecular meshwork (TM) cells in the outflow pathways of eyes of glaucomas of diverse etiology, it was absent in the outflow pathway of all normal eyes examined. We propose a unique hypothesis that TM cell surface E-selectin, by binding to its putative glycoprotein counterreceptors on adjacent cells or in aqueous humor activates key signaling pathways, which in turn, by promoting cell-matrix and/or cell-to-cell interactions and cytoskeletal changes, provide a protective stress response specific to the aqueous outflow pathway of the eye. In this application, we propose pilot studies to determine whether the hypothesis merits a detailed investigation.

In Aim 1, using immunohistochemical and Western blot analysis, we shall establish which specific carbohydrate ligands of selectins, if any, are expressed on the TM cells and in the aqueous humor of normal and glaucomatous eyes.

In Aim 2, we shall determine whether the selectins/carbohydrate-mediated signaling promotes adhesion of TM cells to a variety of extracellular matrix molecules in vitro. The proposed studies will determine whether the selectin-based carbohydrate recognition system plays a role in the pathogenic mechanisms of aqueous outflow resistance in glaucoma.

PI: Papas, Athena
Title: Prevention of Adult Caries (PACS)- Tufts Clinical Center
Abstract: Adult dental caries, including both coronal and root caries, is an infectious disease that afflicts the majority of Americans aged 55 and older and is the most common chronic disease at midlife. In addition, adult caries exacts a significant economic toll, and this economic toll continues to grow. Despite the high prevalence and bacterial pathogenesis of caries, no FDA-approved anti-microbial treatment for caries is available to the American dental professional. The Prevention of Adult Caries Study (PACS) is a randomized, double-blind, placebo-controlled Phase III clinical trial conducted under FDA Investigational New Drug license #45,466. This study is designed to evaluate the efficacy of a topical, temporary, 10% w/v chlorhexidine dental coating in reducing caries increment in at-risk adult dental patients. This study will follow 1000 patients over a 13- month study period at four centers with vastly different populations. The primary endpoint for this study will be caries increment. The centers participating in this proposed research are: Center for Health Research, Kaiser Permanente Northwest, in Portland, Oregon; Tufts University Dental Clinic in central Boston, Massachusetts; Dental Service of Massachusetts Clinical Center in Southborough, Massachusetts, and; the dental clinic of the Tuba City Regional Health Care Corporation in Tuba City, Arizona. Kaiser Permanente's Center for Health Research in Portland, Oregon will act as the data-coordinating center, and Tufts University will act as the Administrative Center. This application from the Tufts Clinical Center is part of the coordinated set of applications in support of the PACS study. Please refer to the Administrative Center Application for a complete description of the study Protocol and Manual of Procedures.

PI: Papas, Athena
Title: Prevention of Adult Caries (PACS)- Tufts Administrative Center
Abstract: Adult dental caries, including both coronal and root caries, is an infectious disease that afflicts the majority of Americans aged 55 and older and is the most common chronic disease at midlife. In addition, adult caries exacts a significant economic toll, and this economic toll continues to grow. Despite the high prevalence and bacterial pathogenesis of caries, no FDA-approved anti-microbial treatment for caries is available to the American dental professional. The Prevention of Adult Caries Study (PACS) is a randomized, double-blind, placebo-controlled Phase III clinical trial conducted under FDA Investigational New Drug license #45,466. This study is designed to evaluate the efficacy of a topical, temporary, 10% w/v chlorhexidine dental coating in reducing caries increment in at-risk adult dental patients. This study will follow 1000 patients over a 13-0month study period at four centers with vastly different populations. The primary endpoint for this study will be caries increment. The centers participating in this proposed research are: Center for Health Research, Kaiser Permanente Northwest, in Portland, Oregon; Tufts University Dental Clinic in central Boston, Massachusetts; Dental Service of Massachusetts Clinical Center in Southborough, Massachusetts, and; the dental clinic of the Tuba City Regional Health Care Corporation in Tuba City, Arizona. Kaiser Permanente's Center for Health Research in Portland, Oregon will act as the data-coordinating center, and Tufts University will act as the Administrative Center.

PI: Park, James
Title: Muropeptide Recycling Pathway and BETA Lactamase Induction
Abstract: Amazingly, Escherichia coli degrades over 60% of the murein (peptidoglycan) from its sidewalls each generation and is able to reutilize the components. This process is known as recycling. The goal of this research is to identify and characterize all the enzymes used by E. coli to reutilize cell wall peptidoglycan components. Recycling involves over 10 enzymes that appear to serve no other purpose than to facilitate breakdown and reutilization of murein components. Several outstanding questions remain to be answered in order to have a full understanding of the process. The recycling pathway is required for beta-lactamase induction in many Gram negative organisms and, in addition, we have recently observed that mutations in certain recycling genes result in auto aggregation and biofilm formation. Thus the pathway is of interest in the health related areas of resistance to penicillins and biofilm formation.

Specific aims:

Aim #1: Determine the pathway for utilization of N-acetylglucosamine in the absence of the N-acetylglucosamine kinase. We have already achieved the original goal of Aim 1, namely, Identify the kinase gene and demonstrate its role in reutilization of N-acetylglucosamine.

Aim #2: Determine how E. coli degrades anhydro-N-acetylmuramic acid. Our recent results indicate that a kinase and an etherase may be involved. Hence our current efforts are aimed at purifying these activities in order to identify the genes responsible for the activities.

Aim #3: Determine the true inducer of beta-lactamase.

Aim #4: Determine how expression of antigen 43 is related to the recycling of murein tripeptide. Antigen 43 causes auto aggregation which facilitates biofilm formation. Deletion of either murein peptide ligase (mpl) or murein peptide amidase A (mpaA) results in production of tripeptide and antigen 43 suggesting that accumulation of the murein tripeptide, L-Ala-gamma-D-Glu-meso-diaminopimelic acid, may be involved.

PI: Perrin, Mercio
Title: Neuron Survival in Chagas Disease
Abstract: Chagas disease, caused by the protozoan Trypanosoma cruzi, progresses through 3 stages:

1. Acute disease, characterized by robust parasite growth, immunological disturbances, and peripheral neural degeneration

2. Indeterminate phase, asymptomatic and featuring extensive regeneration of neurons

3. Chronic phase, characterized by immunological alterations and pronounced degeneration of neurons in the heart and GI tract

Most patients remain asymptomatic and with signs of neuronal regeneration for decades, but for enigmatic reasons, some (<10 percent) undergo extensive degeneration of neurons (along with immunological disturbances) to progress to the fatal chronic disease. Recent studies may provide insights into the molecular basis of neuronal changes in Chagas disease. Picomolar levels of the T. cruzi trans-sialidase (TS) were found to protect several types of neuronal cells from undergoing apoptotic death. What's more, TS synergized with two human neurotrophic cytokines, ciliary neurotrophic factor (CNTF) and leukemia inhibitory factor (LIF), to promote neuronal survival. Therefore, the possibility exists that neuronal regeneration in Chagas disease results from the collaboration of TS with CNTF or LIF. In addition, and most interesting, because about 2.5 percent and 25 percent of normal people are homozygous and heterozygous, respectively, for a null mutation in the CNTF gene, the possibility exists that individuals bearing mutation of this neurotrophic cytokine are more prone to develop extensive degeneration of neurons if infected with T. cruzi and thus to progress to chronic Chagas' disease.

PI: Perrin, Mercio
Title: Receptors for Neuron and Glia Survival in Chagas Disease
Abstract: Trypanosoma cruzi, the agent of Chagas disease, makes an enzyme, trans-sialidase (neuraminidase) (TS), located both on the surface of invasive trypomastigotes and in the extracellular milieu. A most surprising and novel aspect of the TS physiology is its mimicry of neurotrophic factor(s) of mammalian hosts. In neuronal PC12 cells, TS induces neurite outgrowth and suppresses apoptosis without or in synergy with ciliary neurotrophic factor and leukemia inhibitory factor. In glial Schwann cells, it functions as a specific survival factor. TS-induced survival of PC12 and Schwann cells requires intact phosphoinositol 3-kinase (PI3K)/Akt kinase signaling. These TS actions are potentially relevant to the pathogenesis of Chagas' disease. However, it is unknown how TS does it. Identifying and cloning neuronal and glial receptors for the trypanosomal neuraminidase should aid in understanding the precise mechanism of TS activation and in identifying the cells on which TS acts. Receptor identification should also provide essential tools with which to further ascertain the biological significance of TS signaling in the nervous system that becomes infected with T. cruzi. One practical aspect that could result from such work is the possibility of developing reagents to treat Chagas' and other neurodegenerative diseases. We propose the following approaches to characterize TS receptors on cells of the nerve system:

Specific Aim 1: Developing, by genetic engineering, an enzymatically inactive TS fragment for determining TS binding parameters to PC12 cells, Schwann cells and astrocytes.

Specific Aim 2: TS ligand blotting for trkA and other relevant neurotrophic receptors.

Specific Aim 3: Tagged-ligand affinity chromatography

Specific Aim 4: Expression cloning by tagged-ligand panning

Specific Aim 5: TS-induced activation of TS-binding proteins: phosphorylation of trkA; activation of the nuclear factor КB (NF-КB), a downstream signaling of p75neurotrophin receptor; and other mechanisms.

PI: Peterson, Julia
Title: Citrus Flavonoids to Promote Health through Decreasing Cancer Risk
Abstract: There is some evidence that citrus fruits not only promote general nutritional health but also decrease risks of human cancers. However, the specific fruits and the compound or compounds associated with these effects have not been identified. Therefore, plant-breeding efforts cannot be directed toward developing citrus varieties and products rich in the bioactive compounds that may have these effects. And, tailoring dietary advice or recommendations for citrus foods or supplements that will improve the Nation's nutrition and health and enhance economic opportunities for agricultural producers is difficult. The hypothesis is that increased intakes of the flavanone and flavone classes of flavonoids in citrus fruits are associated with decreased risk of breast, colorectal, and head-neck cancers. Nine epidemiologic case-control studies will be used to economically and rapidly evaluate the effects of these citrus constituents on cancer risk. Comprehensive flavanone and flavone data including additional citrus flavones (diosmetin, nobiletin, sinensetin, tangeretin) will be developed and used to analyze these data, improving completeness of exposure assessment. Available datasets consist of approximately 3,400 cases of breast cancer, 2,000 of colorectal cancer and 1100 of head-neck cancer (700 oral and pharyngeal and 400 esophageal) with a similar number of controls. The case-control studies will be analyzed using multivariate analysis and logistic regression with modeling to control for possible confounders. This project will develop new flavonoid data on the flavanone and flavone classes, explore if citrus flavonoid intakes decrease colorectal and head-neck cancer risks, and confirm or refute preliminary observations of flavonoid intake and decreased breast cancer risk. This project will contribute new data to the expansion and upgrading of the United States Department of Agriculture's publicly available flavonoid food composition database, facilitate the work of other investigators, and assist citrus producers in the development of products that promote health.

PI: Pinderhughes, Ellen
Title: Multisite Prevention of Adolescent Conduct Problems
Abstract: This proposal is to complete the evaluation of a developmentally based, long-term, comprehensive intervention designed to prevent serious antisocial behavior and related adolescent problems. The project is being carried out at four sites (Durham, NC/Duke University; Nashville, TN/Tufts University-Vanderbilt University; rural Pennsylvania/Pennsylvania State University; and Seattle, WA/University of Washington). The screening procedure of Fast Track identified three successive cohorts of high-risk children in kindergarten by their early conduct problems at home and school. These children were randomly assigned by school to an intervention or control group. Intervention began in first grade with high-risk children, their adult caretakers, and their teachers based on a developmental model that specified six areas of risk and protective factors: parenting, child problem-solving and emotional coping skills, peer relations, classroom atmosphere, academic achievement, and home-school relations. It was continued in adolescence with an emphasis on protection from deviant peer influence; promoting positive identity, goals, and aspirations; and academic and vocational skill development. Analyses indicate the intervention has led to significant improvements in the hypothesized risk and protective factors and reductions in conduct problem behavior over the elementary school years. The primary aims of this proposal are:

1. To complete the assessments in the last 2 years of high school and the 2 years immediately after high school in order to evaluate intervention effects on mental health, crime, substance abuse, education and employment, and the use of community services
2. To determine whether characteristics of the participant sample moderated the effects of the intervention
3. To understand factors that mediate successful preventive intervention
4. To identify factors influencing participation and the relation between dosage and outcome
5. To continue to test the developmental model of early and late starting conduct problems with the normative and high-risk samples
6. To continue to develop innovative methods for analyzing data from prevention trials
7. To provide data for an economic study of the impact of Fast Track on professional service utilization

The data collection plan calls for completion of annual parent and youth assessments at the end of 11th (cohort 3) and 12th grades (cohorts 2 and 3); a phone interview 1 year after the end of high school; and a face-to-face interview at age 20 that will provide a final assessment of mental health status, conduct problem behavior, substance use, educational progress, and employment status at the end of the teenage years (all cohorts).

PI: Pinderhughes, Ellen
Title: Adoption and Development
Abstract: The specific goals of this study are to:

• Examine the identity and understanding of adoption of girls adopted from China who are being raised by families in the U.S., as well as their experiences associated with being Chinese-American adoptees

• Examine parents’ experiences raising their Chinese-American adoptees, and family functioning

• Evaluate the impact of the playgroup on the girls’ understanding of adoption and their ethnic identity

This study will serve as a pilot study for a larger grant submission to NICHD that will follow-up these participants and include other Chinese-American adoptees and their families. Nationally, families adopting internationally are quite diverse with respect to parental constellation (two parent heterosexual, two parent homosexual, single parent). Most, however, are European American and middle income families. How do girls raised in these families view their ethnicity and understand adoption? The following questions will be examined:

• What are the experiences of girls adopted from China and raised in the U.S.?
• What are their experiences with peers and how do they perceive these experiences?
• What experiences do their parents provide and how are the adoptive parents dealing with the issue of raising children who are ethnically different from them?
• How does a play group in which the girls have participated since age three facilitate their understanding of difference, adoption and their cultural background?

PI: Pokras, Mark
Title: Training Tomorrow’s Leaders: Energizing a New England Conservation Medicine Network Through Innovative Student Training
Abstract: Thinking About Animals and Environmental Health

1. Crows and other wild birds firstsignaled that West Nile Virus had been introduced to the Americas
2. Sick and dying ducks in Boston and Worcester are proven early indicators of botulism and Cryptosporidium in bodies of water used by the public
3. Pet dogs and cats can indicate health threats to people including lead poisoning, cancers, and fungal diseases
4. Some New England birds are showing altered behavior that may be attributable to elevated levels of mercury and organic chemicals in fish they eat. Many are the same fish that people consume, especially in impoverished, rural and immigrant communities
5. Marine mammals along the New England coast are dying of morbilliviruses (a new disease related to diseases of domestic animals and people), and toxic algal blooms

To investigate and solve such complex problems requires the combined expertise of professionals trained in a variety of disciplines. As noted historian Theodore Zeldh commented, “Most advances in science have been the result of intermediaries venturing beyond the boundaries or the paradigms of their disciplines, uniting insights which come from different kingdoms of knowledge."

PI: Pokras, Mark
Title: Massachusetts Oral Rabies Vaccination Program
Abstract: Tufts Cummings School of Veterinary Medicine (TCSVM) established The Cape Cod Oral Rabies Vaccination Program (CCORVP) in 1993 with funding from the Massachusetts State Legislature, in cooperation with the Massachusetts Department of Public Health (MDPH), the Centers for Disease Control and Prevention (CDC), and USDA Wildlife Services joined the project in 2001.

The original goal of the project was to prevent the spread of rabies onto Cape Cod. For 10 years, from the first vaccine distribution in May 1994 to March 2004, TCSVM and our cooperating partners were successful in keeping raccoon rabies off Cape Cod despite epizootic challenge. We also succeeded in achieving the highest vaccination rates in raccoons for any vaccine program in the country, and establishing vaccination strategy techniques for a suburban landscape that have been used nationally.

In March of 2004 a breach in the vaccine barrier to Cape Cod was identified. This breach appears to be due to many factors including the possible role of skunks in the transmission cycle of raccoon rabies. Response to this initial breach in the vaccine barrier was rapid, and included many multi-agency coordinated actions by TCSVM, USDA Wildlife Services, MDPH, Barnstable County, local towns and volunteers on Cape Cod. The response included a program to trap, vaccinate, and release raccoons, dispensing oral rabies vaccine in several large targeted areas, enhanced disease surveillance and testing, and educational efforts to inform the public about rabies in wildlife and the promotion of pet vaccination. The Oral Rabies Vaccination (ORV) project will continue to assess the various factors involved in the breach of the vaccine barrier and the spread of the epizootic in order to formulate the best control strategies.

PI: Pokras, Mark
Title: Massachusetts Wildlife Disease Surveillance Project
Abstract: The Massachusetts Department of Public Health (MDPH) will contract with Tufts Cummings School of Veterinary Medicine (TCSVM) to continue the Massachusetts Wildlife Disease Surveillance Project (WDSP). Continuation of the work will further strengthen the capabilities of the State of Massachusetts to guard the public health by monitoring, detecting and responding to potential zoonotic disease outbreaks within the wild animal populations in the state. TCSVM will continue to develop and refine standard operating procedures (SOPS), protocols, inter-agency networks, and other tools to further the development of this program. Emergent diseases, such as avian influenza, with the new potential for grave public health consequences, as well as, re-emergent zoonotic diseases, such as rabies, with a history of public health concerns in the state, will continue to be the focus.

TCSVM will continue to take a leadership role in the development of collaborative efforts statewide for zoonotic disease monitoring and surveillance. We will continue to chair the Animal Surveillance and Education Committee and Avian Influenza subcommittee. We will continue to identify and develop communication and collaboration with agencies and groups in Massachusetts involved in various aspects of monitoring, surveillance and response to wildlife diseases. This will include veterinarians, wildlife rehabilitators, animal control agents (both municipal and commercial), the academic community, state agencies, local boards of health, and others. Because long-standing and functional networks of public health, state and federal agencies and community groups have been developed as part of the Oral Rabies Vaccination (ORV) program on Cape Cod, the WDSP will focus many of its efforts in the geographic region of Barnstable County. The WDSP will use the existing networks and programs to facilitate further development, implementation and testing of the various protocols. By including the broader goals of wildlife disease surveillance in the ongoing work of the ORV, the WSP will benefit from the previously established efforts of public education and community involvement. These efforts will facilitate operational expansion of these surveillance and emergency response procedures and protocols to other areas of the state and to other zoonotic wildlife diseases of concern to human health. In addition, continued ties will be made between this project and the work conducted by USDA-WS in that region to take advantage of their wildlife field sampling as a resource for monitoring baseline levels of other zoonotic diseases such as avian influenza, tularemia and ehrlichia.

The pilot sampling program utilizing the Wldlife Clinic at TCSVM and other sample sources will be continued, allowing the project protocols to be tested and refined. This season's sampling efforts will have two goals: to develop and test the new protocols and SOPS, and to continue gathering background data on target syndromes and diseases. It is anticipated that during this year, the majority of samples obtained and submitted to the MDPH laboratory will be clinical samples (including serum, blood, and feces) to be examined for the specifically targeted infectious agents. A limited number of complete necropsies are anticipated and will be performed at TCSVM.

PI: Poltorak, Alexander
Title: Genetic Dissection of Lipopolysaccharide Response
Abstract: The lipopolysaccharide (LPS) signaling pathway has been analyzed using combination of biochemical and genetic methods. However, not all of the proteins that participate in LPS recognition have been identified. In order to find more of them, several inbred strains of mice were evaluated on their LPS response and genetic basis of the differences in this response was examined. Among them, BALB/C and C3H/HeN strains differ significantly in their response to LPS.

To further determine the relationship between TLR4 LPS response, (BALB/C x C3H/HeN) F2 intercross mice were analyzed on their LPS response and genotyped for TLR4 locus. These data show that although there is functional polymorphism in TLR4 observed between BALB/C and C3H/HeN mice, additional genes are clearly involved in LPS response.

In order to map these genes, the panel of F2 mice was expanded and subjected to genome wide screening with all progeny genotyped for TLR4 locus. Genetic analysis of 80 meioses revealed that hyporesponse of BALB/C mice to LPS is linked to loci on chromosomes 6 and 17. To further characterize candidates and to improve resolution of genetic mapping, two congenic strains were constructed: one with BALB/C allele of TLR4 on C3H/HeN background and another with HeN allele of TLR4 on BALB/C background. These strains were further analyzed to determine the level of their response to LPS.

Two new loci, termed Lpml and Lpm2, may be assumed to contribute to the LPS response. The elucidation of Lpml and Lpm2 will represent an important advance, in that many genes that are known to contribute to complex traits can be identified by such approach.

A similar strategy was employed for analysis of intercross response to peptidoglycan in F2 intercross animals. Elucidation of a precise mechanism of anti-bacterial response in a mouse model will undoubtedly contribute to the studies of host resistance to infectious diseases in humans.

PI: Pothos, Emmanuel
Title: The Effects of Dietary Obesity on Midbrain Dopamine Systems
Abstract: Despite recent epidemic proportions of dietary obesity, there is a considerable lack of knowledge about the neuro-chemical effects of obesity on those midbrain dopamine projections involved in motivation and food reward. A fundamental issue is whether the effects of dietary obesity on CNS function are due to changes in body weight and bodily metabolism; or due to the palatability of high-energy diets, which could significantly alter synaptic plasticity in midbrain dopamine systems and change their response to dopamine-releasing agents, such as food or drugs of abuse.

In this proposal, we will consider the hypothesis that chronic exposure to a cafeteria-type palatable diet (leading to excessive weight gain) significantly reduces basal and evoked dopamine release in the adult rat nucleus accumbens and other targets of the midbrain dopamine cell bodies (such as the medial prefrontal cortex and the striatum). We also predict that this attenuation in basal and evoked dopamine release would reduce the dopamine-releasing efficacy of palatable meals and induce obese animals to increase the intake of palatable food or other non-physiological dopamine-releasing reinforcers (like psychostimulants) in order to achieve dopamine release comparable to that of a normal weight animal. Furthermore, we predict that an increase in evoked dopamine signal in central dopamine systems early in postnatal age is a marker for obesity resistance.

Our specific objectives are as follows:

1. Identify selective effects of dietary obesity (induced by a cafeteria-type diet versus a high-fat diet versus high-carbohydrate diet) on basal, meal and psychostimulant-challenged dopamine neurochemistry in all major pathways of the adult rat midbrain dopamine system (mesoaccumbens, nigrostriatal, corticolimbic) with in vivo microdialysis. Correlate changes in dopamine release with changes in behavior (locomotion, diet consumption, response to injections of the opiate antagonist naloxone). Compare results with those on rats inbred for several generations to be either obesity-prone or obesity-resistant.

2. Identify selective effects of dietary obesity on electrical stimulation-evoked dopamine release in real time in all major pathways of the adult rat midbrain dopamine system (mesoaccumbens, nigrostriatal, corticolimbic) with acute slice amperometry. Study differential effects of obesity on neurotransmitter release versus reuptake. Compare slices from normal weight, dietary obese, inbred obesity-prone and inbred obesity-resistant animals.

3. Use cultured ventral tegmental area dopamine neurons from inbred dietary obesity-prone and obesity-resistant neonatal rats to consider whether dopamine quantal release is higher in obesity-resistant animals before actual exposure to palatable diets and excessive weight gain.

PI: Quinto, Eric Todd
Title: Tomography and Integral Geometry
Abstract: The first objective is to solve problems in tomography. Professor Quinto will develop and refine algorithms for industrial limited data tomography. He will develop algorithms for SONAR and geophysical testing, and he will continue working on radiation dose planning. Professor Quinto has tested his limited data tomography algorithm on real industrial data; these tests were successful, and he will refine the algorithm using these results. He will also numerically test singularity detection algorithms for this problem. Professor Quinto will develop a boundary detection method for SONAR in shallow water and geophysical examination of near-to-surface structures. The theoretical underpinnings are based on theorems of Professors Agranovsky and Quinto about spherical Radon transforms. The algorithm will be tested on simulations using different physical models. Professor Quinto will work jointly with Professor Borgers on medical radiation dose planning, the goal of which is to use X-rays to kill tumors in the body and to damage as little surrounding tissue as possible. The researchers will characterize the null space of the dose operator for both the semi-discrete and completely discrete problems. They will investigate the dependence of the singular values of the dose operator on the attenuation. This applied research uses ideas from Professor Quinto's pure mathematical research.

The second objective is to discover properties of generalized Radon transforms and apply them to other areas of mathematical analysis. Radon transforms have an intimate relation with applications in approximation theory and partial differential equations, and complex analysis, and the results will be applied to these fields. The principal investigator will prove uniqueness and support theorems for Radon transforms defined by spreads of polynomials, including spherical transforms in Euclidean space and the hyperbolic plane and spreads defined by other polynomials in Euclidean space. He will use these results to answer questions in approximation theory and partial differential equations. He will prove properties of the X-ray transform in Euclidean space. He will prove support theorems for Radon transforms on spheres and other surfaces on manifolds and use these to solve Morera problems on complex manifolds.

PI: Raychaudhuri, Debabrata
Title: Small Molecule Inhibitors of Bacterial Cell Division
Abstract: FtsZ is an essential tubulin-like GTPase that assembles into a ring structure at the site of cell division and recruits other essential division proteins to form the septal ring critical for bacterial cytokinesis. FtsZ is widely conserved in the Bacterial kingdom, including most pathogens and biothreat agents, but is absent in the mitochondria of higher eukaryotes. The essentiality of FtsZ in bacterial cell division, its widespread conservation, its low amino acid identity with tubulin, and its absence in mammalian cells make it an attractive broad-spectrum antibacterial target. Using FtsZ protein-based as well as chemical genetic high throughput screens against small molecule libraries, a number of hits have been identified that cause cell filamentation and bacterial lethality. The whole-cell screens have identified two classes of molecules that cause lethal filamentation with or without affecting FtsZ activity, suggesting inhibition of other essential but as yet unidentified septation targets. One goal of the project is to use these division inhibitors as chemical tools to study septal ring assembly and to identify and validate septation-specific non-FtsZ targets. Another goal is to use the promising inhibitors as chemical scaffolds for analog synthesis and structure-function relationship studies. This effort may aid the development of potent therapeutic leads that target cell division in bacterial pathogens of public health importance and in biothreat agents.

PI: Rios, Maribel
Title: BDNF Mutants: Genetic Models for Depressive Disorders
Abstract: Depressive disorders are debilitating conditions that affect millions of individuals and create an enormous burden on society. Close to 100 billion dollars per year are spent treating patients with severe and mild forms of depression in the United States alone. However, the underlying molecular mechanisms that trigger depression remain to be elucidated so that treatment alternatives for patients that are unresponsive to the current forms of therapy can be created.

A potential target for the design of novel treatment strategies is brain derived neurotrophic factor (BDNF). Compelling evidence shows that BDNF modulates affective behavior but the specific role and the mechanism of action of this neurotrophin remain elusive.

We recently generated conditional mutations of BDNF using the cre recombinase/IoxP system. These mice have a pre or postnatal depletion of BDNF in the central nervous system that does not compromise their viability as the global depletion of BDNF does. These mutants display dramatic changes in behavior including hyperaggression and hypersensitivity to stress, both of which are often symptoms of depression.

We propose using these mutants, and others that we are currently generating, as genetic models of depressive disorders to dissect the role of BDNF in the regulation of behavior. Different lines of mutants that through genetic manipulation have depletion or over expression of BDNF in different regions of the brain associated with mood disorders will be tested using standard behavioral models for depression and aggression.

PI: Roberts, Susan
Title: Dietary Energy Restriction and Metabolic Aging in Humans
Abstract: Reducing morbidity and delaying mortality are recognized as major goals of aging research, and are addressed by this proposal to conduct a 2-year human caloric restriction (CR) intervention.

A 1-year pilot study will be conducted in 32 overweight men and women to develop an effective CR regimen when fed at 70% of energy requirements determined at baseline. As part of this pilot we will refine all aspects of a CR intervention, including exercise and behavioral counseling, and will obtain necessary information on outcome variability with which to perform power calculations for the main study. Subjects will be randomized to two dietary regimens with different levels of dietary fat and glycemic index (GI) (20% fat and moderate GI versus 35% fat and low GI) and dietary compliance and key outcome measurements will be determined at 5 periods throughout the year. Dietary factors such as dietary variety, liquid sources of energy, and dietary fiber will then be taken into account in the design of the interventions.

Following identification of an effective CR regimen, a randomized 2-year intervention will be conducted in 117 overweight men and women fed 70%, 80% or 100% of energy requirements determined at baseline. The hypothesis will be tested then, compared to control subjects fed 100% of baseline energy requirements.

The parameters to be evaluated will include immune function, oxidative stress, fasting insulin, hemoglobin Alc, and cardiopulmonary function. W further hypothesize that, compared control subjects, individuals randomized to 70% or 80% of baseline energy requirements will not experience adverse change sin thyroid and reproductive hormones, bone mineral density, disease incidence, mood or cognitive function. Dose-response relationships between the extent of CR and changes in outcome variables are anticipated.

As part of the study, changes in total energy expenditure and resting metabolic rate, body composition and body temperature will be quantified to document the effects of CR on energy metabolism.

We anticipate that the results of this study will have a major impact on our understanding of the relevance of CR to human health. In addition, this study will contribute to the development of new avenues for long-term treatment of overweight and obesity.

PI: Rogers, Beatrice
Title: Water and Health Training Grant
Abstract: In 2002, Tufts University created an interdisciplinary doctoral program to train students to work with a global view toward assuring water security for the protection of health, the environment, and human livelihoods. The program is called Water: Systems, Science, and Society (WSSS), and includes six schools of the university: Medicine, Veterinary Medicine, Nutrition, Arts and Sciences, Engineering, and the Fletcher School of Law and Diplomacy. WSSS is designed to assure that students acquire interdisciplinary knowledge and approaches, even as they develop a specialization. With this application we propose to support students in WSSS with a concentration in Water and Health. Problems of water needs, use, and quality are pervasive throughout the world as well as the US, and are considered important enough by the National Institutes of Health to be mentioned specifically within the goals of Healthy People 2010. The Water and Health training program will support both HP 2010 goals and the New Roadmap goals of creating a new kind of interdisciplinary professional. Integrated, interdisciplinary analysis is necessary to understand and manage complex water related problems. Our vision is to educate professionals skilled in health-related disciplines who can use multiple disciplinary perspectives and tools. Water and Health graduates will be able to respond to the interdisciplinary, integrated water-related research challenges. Engineers will understand the principles of Public Health. Nutrition students will understand the basics of hydrology. Policy students will understand the fundamentals of epidemiology. These new professionals will be able to participate in the analysis of water problems and work to develop solutions jointly with colleagues from many specialties, using the rigorous methods and tools of the relevant disciplines.

Rogers, Beatrice
Title: Hunger Atlas Follow-up Analysis
Abstract: Hunger and malnutrition are at the center of economic development. Malnutrition is an indicator of poverty, social exclusion, and deprivation, and is a major constraint to national economic development. For this reason, the reduction in hunger is part of the first Millennium Development Goal (MDG).

The Nutrition Analysis and Mapping Project, jointly developed by the World Food Program and Tufts University, was undertaken to assist in identifying areas of high malnutrition prevalence at a geographically disaggregated level, to assist in understanding the determinants of nutrition problems in different geographic settings and in targeting nutrition interventions, and to provide a basis for monitoring progress in the achievement of the first MDG.

The output is a series of maps that show, at the municipio level, the estimated prevalence of childhood chronic malnutrition (low height for age of children 6-59 months), and that demonstrate associations with other geographic and economic characteristics and with the provision of services.

After completing the pilot exercise in Dominican Republic, it is necessary to carry-out additional analysis to ensure that the methodology developed can be transferred and replicated in other countries in the region, as well as a quality control tool for the first pilot exercise. Another important aspect of this phase will be the integration of the VAM into the methodology, as well as explore potential complementarities.

PI: Rogers, Chris
Title: Teaching Through Touching: Using Research to Motivate Education
Abstract: Engineering education can be made exciting and attractive to students of all backgrounds, races, and gender if it includes the excitement of open-ended research. That is, if neither student nor teacher knows the “right answer” to the question. Unfortunately, this kind of engineering or scientific inquiry rarely happens before graduate school.

Over the past 15 years, Professor Rogers has been working to change that. During this time, Professor Rogers has built up an internationally recognized program in particle-laden turbulence, in chemical-mechanical planarization (CMP), and in engineering education. His work in education has led to multiple international alliances aimed at bringing engineering into the liberal arts curriculum in universities, into the K-12 classroom, and in improving engineering education for the college engineering student as well.

At the core of these efforts is the desire to teach through research, giving students of all ages the tools and self-confidence to answer their curiosity and to understand math and science. Over the past 5 years, the PI has developed the ROBOLAB™ toolset, a combination of LEGO™ hardware and LabVIEW ™ software that allows students to predict, investigate, test, and understand math, science, and engineering. ROBOLAB is designed to have a very low entry (youngest user was a 3 year old) and high ceiling (it still poses challenges for graduate students).

This proposal has three specific goals:

1. To investigate how students learn engineering (both the kindergartner and the university engineer)
2. To push the ceiling of the PI’s toolset higher by combining the robotics toolset with the CMP research
3. To increase the collaboration of all institutions (K-12 and university) dedicated toward bringing open-ended research problems into the classroom as a teaching method

This work would be the primary focus of a graduate student team, one in education and one in engineering and a faculty team, some in engineering, some in education. We would involve universities, industry volunteers, government institutions, and science centers around the world, supplying a support and dissemination structure for these efforts.

We would try new directions in both research and education, for instance integrating the virtual world with the real world of ROBOLAB. We would test new informal education efforts, including integrating math, science, and engineering education with movie cinemas and the Internet. Professor Rogers’ long-term goal is to increase the engineering literacy of the average graduate, reducing the phobia associated with engineering and breaking down the walls between engineering and the liberal arts courses. To do this effectively, one must start in the kindergarten and continue the effort through college.

This proposed work would provide a big step toward that goal by helping in the dissemination of the engineering material to that K-college body, by helping understand the cognitive processes behind the learning, and by ensuring that the toolset is flexible enough that all students are challenged.

PI: Rogers, Chris
Title: Feature Scale DELIF Investigations in Chemical Mechanical Planar
Abstract: Over the next three years we propose to run a set of experiments to investigate better how the pad behaves under the wafer during CMP. In particular, we will look at pad behavior under 1mm square (10-40 micron deep) wells etched into the surface of the wafer. All measurements will be taken during polish with both the wafer and pad rotating. The white spots are regions with more slurry between the wafer and pad and, conversely, the dark spots are areas of less slurry. In this way we can see individual asperities in the pad and determine what happens around topography on the wafer. Some critical questions that we would like to answer are:

1. How does downforce affect asperity rebound under the well?
2. How does well depth change the pad rebound? What is the critical well depth, after which the pad completely relaxes under the well?
3. How do sub-pads affect pad rebound?
4. How quickly does the pad rebound? This would be a spatial measurement — that is to detect well do the pad asperities completely rebound?
5. How does pad grooving affect pad rebound?
6. How do different wafer feature shapes affect the slurry and pad behavior?

We propose to combine experimental measurements with a theoretical analysis to build a better model for estimating the pad and fluid behavior under the wafer during polish. This will be the doctoral work of two graduate students, working together. We will also take advantage of the new force table to better correlate the micro-behavior.

PI: Rogers, Chris
Title: Collaborative Research: Telling the Story – Learning Math, Science and Engineering Through Animation
Abstract: The goal of this project is to introduce the concepts of computing and physics to children (and teachers) through the creation of stop-action and 3D-animated movies.

By empowering children with new, usable tools for exploring the world and expressing ideas through animation, and by developing methodologies for incorporating such tools into Science, Technology, Engineering, and Mathematics (STEM) education, this project will broaden the class of students who are not merely exposed to but rather engaged with technology.

This effort leverages emerging public fascination for computer animation, as well as recent technological advances that have moved the graphics power of yesterday’s million-dollar visualization supercomputers into every desktop PC.

A proof of concept of this approach, based on stop-motion animation, has been prototyped by one of the principal investigators. Initial trials have been encouraging: in a high-school physics class for noncollege-bound seniors, students who typically skipped class were now attending, some coming even during free time to complete their movies.

Through animations, students were able to critically examine their own understanding of the physics and more effectively convey that understanding to teachers. The same technique is also being used to teach reading to 7 year olds and biology to 9 year olds, replacing book reports and lab notebooks with animated stories and documentaries.

Informed by this experience, the proposed program will have two arms: one seeks to develop and evaluate teaching methodology based on moviemaking (Tufts University), while the other will advance new 3D computer animation tools useable in the classroom (Princeton University).

Intellectual Merit: New and potentially powerful technological teaching tools are often developed in the absence of strong educational research. This project will use accepted metrics (and develop new ones) to quantify the STEM learning improvement in high school physics as a result of using animations, comparing student understanding in conventional “hands-on” physics classes with those that include movie journaling.

Results from this work will not only contribute to our understanding of how students learn physics and computing, but also help bridge the student’s experience and intuition with modern scientific theory. Further development of moviemaking tools will allow students to move from the jerky animation of the stop-action world to the smooth animations of modern computer graphics. Unfortunately, existing animation systems are barely usable by professionals, let alone grade-school students. This proposal will address this research challenge by developing inexpensive and robust 3D scanning hardware, point-and-click animation interfaces, and methods for stylized (e.g. cartoon-like) rendering of 3D animation.

Broader Impacts: Anecdotal evidence from the prototype system (gathered over the last three years in five classrooms) already suggests the potential significant impacts of the work. Science-phobic students and computer-shy teachers enthusiastically argue about the underlying physics to improve their movies. Movie making gives teachers a multi-media portfolio to assess student learning and test student preconceived models.

If formal evaluations agree with this experience, the results of this proposal have the potential to change the way students learn science at all ages, opening up a new channel to students to show their understanding and test their hypotheses. This may lead to innovations in teaching computing, math, biology, chemistry, engineering, and even story telling and literature. (Nonetheless, this study chooses an emphasis on physics education because of established metrics for evaluation in this subject.) Even more broadly, animation represents a new medium of expression—visual rather than written—that is compelling but currently limited to highly skilled professionals.

We propose to develop tools that will make animation more accessible both to children and, more generally, to everyone outside the animation industry. Making this technology more widely available has the potential to affect the way we all communicate, learn, work, and play, turning us into media developers rather than media consumers.

PI: Rogers, Chris
Title: Tufts Engineering the Next Steps (TENS) GK 12 Project
Abstract: In this application for an accomplishment-based renewal of the Tufts' Engineering: the Next Steps (TENS) GK-12 project, we report the impact that the program has had both on area schools and on other GK-12 programs.

Our long-term goal is to increase pre-college engineering education on a national scale and see the GK-12 model as one successful way of accomplishing this. We feel, however, that more research is needed on the impact of the GK-12 program on the attitudes and engineering skills of the participating fellows. In addition, we would gather and disseminate the best practices developed at GK-12 (and GK-12-like) programs across the nation in an effort to increase communication between these groups.

At Tufts we have three strengths that uniquely position us to undertake this project:

1. The Center for Engineering Education Outreach, which has growing expertise in engineering education research
2. Our interactions with GK-12 programs in our region and across the U.S.
3. The successful development of a sustainable outreach program called STOMP (Student Teacher Outreach Mentorship Program) based on the GK-12 model

We have three goals over the next two years. One of these is a local goal, based upon our work with the school system of Malden, MA, which will continue beyond the scope of this grant. The second and third goals propose "deliverables" of national impact. The three goals are:

1. Continue the current TENS project in the Malden school system, using the two further years to complete the transition from GK-12-led to teacher-led engineering initiatives in the classroom.

2. Complete educational research on the benefits of GK-12 work to graduate fellows, and bring the research to national scale with a study using past graduate fellows from all NSF-funded GK-12 programs.

3. Disseminate successful approaches to educational outreach through a fellowship exchange program and through a report of best practices in outreach centered on the GK-12 model.

Tufts’ Engineering the Next Steps (TENS) GK-12 project began work in the summer of 2003 on Track 2 funding from NSF. In many ways, however, it was essentially a new GK-12 program, with a shift in PI, school system, and emphasis. TENS currently works with eight different schools in three urban districts in the Boston Area. While the Malden district remains the main partner district, individual schools in surrounding towns have expressed interest in our program.

Thus, the GK-12 program and STOMP have reached out to some of these schools and has sent over 25 undergraduate and 20 graduate fellows into area classrooms. TENS fellows have worked with more than 65 teachers and approximately 3000 students in the various districts. The goal of the TENS GK-12 project is to work with K-12 teachers to infuse engineering concepts and skills into their classes. The fellows partner with the teachers for the entire school year to modify lessons that the teachers are currently teaching so they also include engineering concepts.

TENS GK-12 is housed in the School of Engineering at the Center for Engineering Educational Outreach (CEEO). This center is dedicated to improving engineering education for all ages and does this through a number of avenues. These include educational tool development, research on engineering education, and support for teachers and students in K-12 classrooms. The CEEO also has run a Research Experience for Teachers program, which was used as an entry point and recruiting tool for teachers to participate in TENS. The CEEO also runs over 5 teacher related workshops every summer (from NSF funded RET programs and Teacher Enhancement programs to Robotics workshops). We use these for teacher recruitment and match them with Tufts engineering students.

Formal data are still being collected, but anecdotal evidence suggests the undergraduate team has made a significant impact on the classrooms in area schools. For instance, the Medford superintendent has expressed interest in expanded involvement to all schools in the district. Throughout the project, both graduate and undergraduate fellows have worked in conjunction with the teachers with whom they are partnered to produce effective lessons and hands-on activities. Fellows concentrate on integrating engineering projects with existing curriculum so that GK-12 related activities support State Curriculum Frameworks rather than displace existing content. The lessons deemed effective by the teacher/fellow teams are being collected in a single database so that documentation and supporting materials for these lessons can and will be disseminated to all teachers through the Malden and Medford Science Coordinators. These activities will also be available to the rest of the nation on the Web at the TENS and CEEO websites during the summer of 2005.

Sustainability:  We have developed two different sustainability models for the GK-12 concept that will exist at least 10 years beyond the lifetime of the grant. The first program is the STOMP that has been successfully running at the Center for three years. STOMP fellows serve in a similar role to GK-12 fellows, but the program constitutes a less intensive commitment by the fellows, and pays an hourly rate rather than a stipend. Also, decreasing the fellows’ time in the classroom, while still providing support, will encourage teachers towards taking ownership of the engineering lessons used in their classrooms.

Funding for STOMP has been secured for the next 15 years from the LLL Foundation. STOMP currently places around 20 undergraduates and graduates into the classroom every week in schools ranging from inner city schools to suburban schools. It is completely student run and has caused a noticeable increase in retaining engineers at Tufts. It also has a disproportionately large percentage of female participation. We are currently working with Raytheon, Lockheed Martin, and National Instruments to start a number of industrial STOMP programs, placing their engineers in classrooms to help their teachers teach engineering.

The second program of institutionalization was to create Outreach Assistantships. Similar to a Teaching Assistant or Research Assistant, an Outreach Assistant is paid a monthly stipend to go into the classroom for 5-8 hours per week. In this model, stipends are paid for by a cooperative agreement between the CEEO and the research advisor, with each providing 50% of the funds. This allows for continued outreach work with increased faculty involvement in CEEO activities. Much of this support comes from investigator-initiated research grants that include support for outreach as part of the broader impact of the work. CEEO currently funds two such positions and the funding comes primarily from royalties from ROBOLAB, a popular educational software developed at the Center.

GK-12 Collaborations:  We began building stronger ties with other GK-12 programs in Massachusetts by co-organizing the January 2004 Boston Regional GK-12 Regional Conference. The Conference proceedings were disseminated on the web, and was distributed on a CD to everyone in attendance. The CD has since been copied and distributed to almost every GK-12 project in the country. Similar Regional Conferences have since taken place in Florida and are being developed in New Hampshire and other regions.

As our program matured, we have begun a student exchange program with other GK-12 (and GK-12-like programs) around the country. Last year was our first exchange with the Columbia University “Track 2, GK-12: Technology Integration Partnerships: Bringing Emerging STEM Research into Grades 5- 12 Enabled by New Technologies” project. As a result of this exchange, the Columbia program has increased its robotics portion of their outreach. We are currently planning a similar exchange this summer with the Phystec program at the UC Boulder Physics department. With this exchange we hope to create stronger ties to the physics education community. We are also currently talking with GK-12 programs at the University of Missouri and Clarkson.

Finally, we produced the TENS GK-12 Graduate Fellow Manual, which has been requested by more than 15 other GK-12 programs. The manual provides a comprehensive summary of the project and its logistics, school district and individual school information, Tufts University information, information on the surrounding area for people new to the area, and a complete list of TENS policies.

PI: Romero, Michael
Title: Quantifying Physiological Stress in Threatened and Endangered Species Due to Military Activities
Abstract: Organisms must respond to unpredictable, novel, and/or dangerous
conditions in their environment to maintain homeostasis and optimize fitness (survival and reproduction). Response of wildlife populations to human disturbance has long been of concern to ecologists, which is reflected in strong federal regulations (e.g. the Endangered Species Act of 1973, as amended) requiring federal agencies to assess the effects of their actions on listed species. Physiological indicators of stress have been suggested as surrogate measures of human-caused stress in species of concern, notably measures of energy expenditure and the adrenocortical response. Human-related stressors characteristic of military operations that are episodic, unpredictable and varying in intensity have received relatively little attention in terms of physiological response. Mission critical military training and testing have been impacted by known, unknown, or potential impacts on endangered species. This study will evaluate physiological indicators of stress in two endangered avian species, the black-capped vireo and golden-cheeked warbler, to determine sensitivity and plasticity of response to military training disturbance.

The objectives of this proposal are to:

1. Develop “dose/response” models for physiological response of selected priority endangered species to military stressors

2. Determine capacity of species of concern to habituate to “non-threat” disturbances

3. Test predictive models for physiological stress response based on life history characteristics and taxonomic affiliation of endangered species

Meeting these objectives will fill knowledge gaps on the threshold level of disturbance necessary to elicit a physiological response and modulation of this response to repeated disturbance, and will provide a predictive framework for physiological response to disturbance based on life history characteristics and taxonomic affiliation.

PI: Romero, Michael
Title: Physiology of Stress in Wild Animals
Abstract: The vertebrate stress response helps protect an organism from an external noxious stimulus such as (predators or storms). One of the hallmarks of the stress response is the activation of the hypothalamic-pituitary-adrenal (HPA) axis, culminating in glucocorticoid release several minutes after initiation of a stressful stimulus. Glucocorticoids then proceed to induce a variety of behavioral and physiological effects. Although stress has attracted much interest of late, we still .poorly understand how glucocorticoids allow wild and free-living animals to cope with and survive noxious environmental stimuli.

Recent work has begun to examine the role played by glucocorticoids in the stress response of wild animals (Wingfield, 1994). Particularly intriguing are recent studies indicating that wild free-living birds can seasonally modulate glucocorticoid secretion. In other words, the magnitude of the stress response appears to depend upon the time of year. Modulation of glucocorticoid release has far-reaching implications for both the physiology of the stress response and the short- term survival of the individual animal. Glucocorticoids seem to be essential for survival (Darlington et al., 1990) yet some species essentially fail to secrete glucocorticoids during some seasons with no apparent detriment. This immediately raises the question of how animals can survive noxious stimuli without releasing glucocorticoids. Understanding how and why glucocorticoid release is modulated is thus fundamental to our understanding of the stress response.

Furthermore, seasonal modulation of glucocorticoid release may also provide insight into physiological solutions to problems faced by wild animals in natural conditions. Early examples of seasonal modulation were discovered in birds living in the deep desert and the arctic (Wingfield et al., 1992; Astheimer et al., 1995) ─ harsh and unpredictable environments. From a human perspective, these habitats should be very stressful. To individuals of species accustomed to these environments, however, encountering harsh and unpredictable conditions may not be stressful, as long as the harsh conditions occur at normal times and the animal is metabolically prepared. In other words, being adapted to an environment may mean that the environment is no longer stressful. If this is so, then seasonal modulation of glucocorticoid release may not be restricted to species living in extreme environments and may represent a common phenomenon. A comparative study is thus indicated in order to ascertain whether seasonal modulation of glucocorticoid release is a general phenomenon. If seasonal modulation is common, it will provide new information on the ecological validity of how free-living animals respond to stressful environmental stimuli.

To fulfill these research objectives, I will test three hypotheses concerning how and why seasonal modulation of glucocorticoid secretion occurs. These hypotheses are that:

1. Seasonal alteration of glucocorticoid release is common taxonomically

2. That different species use different mechanisms to alter glucocorticoid release

3. That altered glucocorticoid levels are necessary to allow the normal seasonal progression of physiological changes

Testing these hypotheses will entail defining physiological mechanisms controlling seasonal modulation and, equally important, identifying whether temperate avian species show this phenomenon and begin to explore why. At the end of this study we will have a broader idea of what role glucocorticoids play in the stress response, and therefore the survival, of wild animals. Furthermore, we will have a deeper understanding of the neurophysiological control mechanisms that regulate glucocorticoid release in wild animals.

An integral part of this proposal is also to allow students to participate in the research. Engaging students in scientific research is an established method for both enhancing student science learning and for providing breadth to the research. With this proposal I will integrate graduate and undergraduate education into an active research program, thereby broadening our scientific infrastructure.

PI: Rosenberg, Irwin
Title: Renal Transplantation, Homocysteine Lowering and Cognition
Abstract: This application is for an ancillary study to determine the cognitive effects of homocysteine Iowering, as an additional outcome in an ongoing, randomized, controlled, double-blind clinical trial in renal transplant recipients (RTRs).

Elevated plasma homocysteine levels are associated with diminished cognitive function in the general population, and significantly increase the risk of vascular disease, cerebrovascular disease, stroke and dementia. Hyperhomocysteinemia is a pervasive feature of chronic renal insufficiency, even after a successful transplant. However, unlike in other types of chronic renal insufficiency, homocysteine levels can be lowered in RTRs by high doses of B-vitamins.

The parent trial is designed to determine the effect of lowering plasma total homocysteine levels on atherosclerotic cardiovascular disease outcomes in chronic, stable RTRs. The FAVORIT trial aims to randomize 4000 RTRs with a stable functioning renal graft of > six months to a treatment or placebo group, and to follow them for 4 years or until the occurrence of a cardiovascular event or death. Treatment consists of a standard multivitamin with additional high dose folic acid, vitamin B12 and vitamin B6 and placebo consists of a multivitamin devoid of these vitamins.

This application specifically aims to:

1. Determine the cognitive effect of homocysteine lowering under this treatment regime
2. Characterize cognitive function in relation to homocysteine and other risk factors for vascular disease in this high-risk, non-demented population

To do so we will measure cognitive outcomes in 1000 participants in the FAVORIT trial, at randomization and after a 3-4 year follow up. The outcome of this application may be highly significant in improving health care for RTRs and other groups with chronic renal insufficiency.

Our long-term goal is to identify risk factors for cognitive impairment that can be modified through nutritional intervention or dietary supplementation in order to reduce the incidence of cognitive decline and dementia in elderly and other vulnerable populations.

Demonstrating the cognitive benefits of lowering homocysteine may pave the way to nutritional modification of cognitive decline in RTRs and other high-risk groups and even in the general population.

PI: Rosenberg, Naomi
Title: RNA - Tumor Virus Hematopoietic Cell Interaction
Abstract: Abelson murine leukemia virus (Ab-MLV) transformation has long been a valuable model to understand the dynamics by which retroviruses containing v-onc genes induce tumors and transform cells. The virus provides a simple genetic tool to examine the way in which signals that stimulate growth, suppress apoptosis and alter differentiation interface with signals from tumor suppressor genes.

Ab-MLV infection of B lineage cells invariably leads to transformation and tumor induction. This is a rapid process: cells infected in vitro begin to divide soon after viral integration and expression of the v-Abl protein, and infected mice succumb 20 to 30 days post infection. Despite this, the road to transformation is not a smooth one. Viral signals that stimulate cell growth and suppress differentiation and apoptosis are countered by a cellular response that promotes apoptosis and erratic growth.

Understanding how the virus counters the cellular response and uncovering the pathways that tip the balance in favor of malignant growth will have broad implications for our understanding of malignant disease and illuminate the features that confer oncogenic properties to Ab-MLV and other v-onc gene-containing retroviruses. In the Ab-MLV system, the p53 pathway is inactivated in most transformants. In some instances, the cells acquire p53 mutations and in others, the p53 regulatory protein, p19Arf is down modulated. We are using the virus to understand how signals from the virus counter anti-tumor effects mediated by the host.

We will investigate four aims:

1. How does p53 influence the outcome of Ab-MLV infection?
2. How do the products of the Ink4a/Arf locus influence Ab-MLV transformation?
3. How do the p53 and Ink4a/Arf loci affect tumor induction in vivo?
4. How is the target cell specificity of Ab-MLV controlled?

PI: Rosenblatt, Michael
Title: Nature of PTH/PTHRP – Receptor Bimolecular Interactions
Abstract: This research proposal is focused on elucidating the interactions of parathyroid hormone (PTH) with its receptor (Rc, the hPTH1-Rc). Formation of the complex between PTH and the hPTH1-Rc leads to a sequence of events, namely hormone binding, Rc activation, and signal transduction, which culminate in expression of hormonal bioactivity. The impetus for this research program comes from a desire to understand:

1. The fundamental nature of molecular recognition between a peptide hormone and its G protein-coupled Rc
2. The mechanism of action of the hormone (PTH) responsible for minute-to-minute regulation of calcium levels in blood
3. The differences in Rc states (conformations) which translate into hormone agonism, antagonism, inverse agonism, etc.

The introduction of PTH as a major new agent for treatment of osteoporosis also focuses attention on the mechanism of anabolic action of this hormone. By gaining insight into the nature of the hormone-Rc complex, the discovery of small molecule PTH-mimetics may be facilitated by structure-guided design in the future.

During the previous grant award period, by integrating photoaffinity scanning, molecular biology, pharmacology, and structural biology (conformational studies of hormone and Rc, and molecular modeling), we succeeded in generating an advanced experimentally derived model of the PTH-hPTH1-Rc bimolecular complex that provides structural detail and reveals some of the dynamics of hormone-Rc interaction.

We are now positioned to take the next major step in mapping the interface of PTH and its Rc, and to extend our studies to identify the shifts in Rc conformation associated with activation. Specifically, we plan to:

1. Improve the resolution of the map of the hormone-Rc interface
2. Study the interaction of PTH ligands covalently tethered to the Rc
3. Investigate the ability of dual-reactive analogs to simultaneously make contact with two sites in Rc
4. Perform "reverse" crosslinking from Rc to PTH
5. Prepare Rc and constitutively active Rc mutants on a large scale for structural studies of antagonist and inverse agonist interaction
6. Use disulfide bridge formation as a probe of Rc states
7. Integrate all the above efforts in a molecular modeling initiative

PI: Rouzine, Igor
Title: Working Models of HIV Persistence and Evolution
Abstract: HIV infection involves many different cell types (infected cells, uninfected cells permissive for virus replication, HIV specific immune cells, and others) which interact with each other and whose numbers, in general, depend on time. Because of interaction among its parts, the system automatically arrives at an approximate steady state coincident with the asymptomatic phase of an HIV infection. The replicating virus population remains quite constant but exhibits considerable genetic variation in time (and among sampled sequences). Our long-term goal is to use mathematical modeling to aid in understanding how such a system can work. Our approach is to (i) select mathematical models of HIV infection based on the criterion that all their predictions agree with the known features of HIV pathogenesis in representative individuals and (ii) formulate predictions for new experiments to test the models. This strategy of "multiple match" has been successfully used in the physical sciences for dealing with complex systems of unknown structure, but it is only being implemented in HIV research. We will apply this strategy to develop models of virus-immune cell interaction in vivo and explain the mechanism of steady state, and to understand the principal factors of evolution of drug resistance and antigenic escape. At this stage of our research, we will put emphasis on follow-up tests of our models in collaborating groups and design of new tests of updated models.

PI: Roy, Ananda
Title: Potential Role of TFII-I in Immunodeficiency
Abstract: TFII-I is an important multi-functional transcription factor that links events to transcription in several genes. TFII-I is constitutively associated with Bruton's tyrosine kinase (Btk), a non-receptor tyrosine kinase that is essential for normal B cell function, as its mutation causes X-linked agammaglobulinemia (XLA) in humans and X-linked immune deficiency (xid) in mice. We propose that TFII-I is an important and novel component in linking Btk-mediated signaling to transcription in B cells. Furthermore, the TFII-I gene gets deleted in William's syndrome (WS) which is a neuro-developmental disorder with multi-system manifestations, including supravalvar aortic stenosis, hypercalcemia in infancy, mental retardation and cognitive defects. Thus, TFII-I appears to be involved in two genetic disorders: William's Syndrome and X-linked agammaglobulinemia (XLA). Knowledge gained from these studies may help us better understand a critical Btk dependent pathway that links B cell receptor mediated signal transduction to B cell specific transcription. These studies may also ultimately help identify potential target gene(s) that are affected by mutations in Btk. Importantly, these studies may establish possible connections between the neuro-developmental disorders (as in WS) and immuno-developmental disorders (as in XLA). Toward a better understanding of TFII-I function in Btk mediated immune response, we will first map the region(s) in TFII-I important for its physical and functional interactions with BTK. We will determine by deletion and point mutation the region(s) in TFII-I that is important for its interaction with Btk, followed by mapping the sites in TFII-I that are tyrosine phosphorylated by Btk in vitro and in vivo by a combination of site directed mutagenesis, phosphopeptide, finger printing, and mass spectrometric analysis. We will also analyze these mutants in functional transient transfection assays. To determine the functions of TFII-I and its biochemical interactions with Btk in B cells, we will employ in vivo transcriptional analysis. To determine the functions of TFII-I and its biochemical interactions with Btk in B cells, we will employ in vivo transcriptional analysis followed by the interaction studies by co- immunoprecipitation and ectopic expression of mutant forms of TFII-I in B cells. We will also stably express wild type and mutant forms of TFII-I, and Btk in B cell lines, and genetically delete TFII-I from chicken B cells. Finally, to ascertain the localization of TFII-I in the absence and in the presence of non-activated versus activated Btk, first, we will co-express various mutants of TFII-I with Btk in COS cells. Subsequently, we will employ freshly isolated primary splenic B cells derived from wild type, xid and Btk-/- mice and study the localization and tyrosine phosphorylation of TFII-I in the absence and in presence of B cell receptor signaling.

PI: Roy, Ananda
Title: Molecular Analysis of Williams Syndrome
Abstract: Williams-Beuren Syndrome (WBS) is a neuro-developmental disorder with multisystem manifestations, including supravalvar aortic stenosis (SVAS), hypercalcemia in infancy, mild to moderate mental retardation, cognitive defects and characteristic craniofacial features. The frequency of this genetic haploinsufficiency is estimated to be 1 in 20,000 live births. WBS is caused by a hemizygous microdeletion of approximately 1.5 MB, spanning 17 genes at chromosomal location 7q11.23.

Despite these observations, we lack a complete understanding of molecular basis for this disorder. Although this multi-system dysfunction with unusual craniofacial, behavioral and cognitive features occurs most likely due to haplo-insufficiency of several genes, rare cases with much smaller deletions have provided clues to identifying specific genes that may be causal to distinctive physical and cognitive defects.

Two of these genes, GTF21 and GTF3 encode the TFII-I family of transcription factors. TFII-I and its relative MusTRD1/BEN exhibit extensive and overlapping expression patterns in a variety of tissues during mouse pre- and post-implantation development. This suggests a functional role for these proteins in early development. These proteins are also abundantly expressed in the hippocampus, a portion of the brain that plays a role in learning and memory, further indicating that they may be causal to some WBS traits.

While much has been learned about the TFII-I's mechanism of action, relatively little is known about how MusTRD1/BEN functions. To better understand the molecular basis for WBS, we propose to dissect the functional role of MusTRD1/BEN. We will proceed to analyze the physical and functional interactions of MusTRD1/BEN with Smad factors, which are critical for a variety of developmental processes. Finally, we will employ RNAi technologies to determine the biological role of this factor in osteoblast differentiation.

PI: Rozanski, Elizabeth
Title: Incidence of Serum Alloantibody in Dogs with a Known History of Pregnancy
Abstract: Blood transfusions in dogs have become an integral part of advanced medicine. Just like humans, several blood groups have been identified in dogs, which are referred to as Dog Erythrocyte Antigens or DEA. A dog negative for a given blood group can produce antibodies following exposure to that specific blood group, which could lead to life-threatening hemolytic transfusion reactions with subsequent transfusion(s). It has been well documented that such antibodies' production can happen following blood transfusions in dogs. In women, pregnancy may result in sensitization of the mother. However it is unknown if a bitch may also be sensitized by pregnancy. Because of the endotheliochorial-type of placenta dogs have, it is thought to be less likely that pregnancy will sensitize the bitch and result in the production of antibodies. However, bitches with prior pregnancies are excluded from blood donor program and cross-matching is highly recommended prior to even a first blood transfusion, in contrast with nulliparous bitches and dogs. The purpose of this research is to determine if bitches can be sensitized and produced antibodies directed against red blood cells following a pregnancy. This knowledge is crucial to decide if special considerations should be given to previously pregnant bitches when in need of blood products and, if on the other hand, those bitches truly need to be excluded of canine donor programs, as it is frequently advertised.

PI: Rubin, Beverly
Title: Does Perinatal Exposure to Bisphenol a Contribute to Adult Obesity?
Abstract: 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.

PI: Russell, Robert
Title: Nutrition Research for Healthy Aging and Prevention of Chronic Disease
Abstract: The objective of this proposal is to obtain funds for the training of PhD scientists committed to academic careers in nutrition research and chronic disease prevention. The importance of training future researchers these two areas follows from our increasing awareness that nutrition is an underlying pathogenic component of many chronic diseases. These include obesity, diabetes and its complications, non-alcoholic Steatohepatitis (NASH), digestive diseases, chronic kidney disease (CKD), hepatic and colon cancer and cardiovascular disease. Although these disorders have traditionally been viewed as pathologies of the middle-aged and/or elderly, it is now clear that appropriate interventions for these conditions need to be initiated decades earlier in the life cycle. Interventions that can delay or prevent chronic disease(s) afford potentially profound public health benefit, both from reduced morbidity and mortality as well as from cost savings. A recent (2000) CDC study reported that obesity alone accounts for 9% ($180 billion) of national health care costs. The rationale for this proposal is based on the firm belief that nutrition is the most significant and tractable environmental factor that can be modified to prevent or delay chronic disease. This proposal therefore seeks funds to train the next generation of nutrition research investigators to address chronic disease prevention at the molecular, cellular, organismal and/or population levels. Support is requested for six predoctoral training slots for each of five years. All trainees are first admitted to a graduate degree program at the Friedman School and, after one year of coursework, are eligible to be admitted to the Training Program. Acceptance into the Training Program is predicated upon outstanding academic and research achievement during the first year. Faculty preceptors on the Boston Health Sciences Campus, all members of Tufts University's Friedman School of Nutrition Science and Policy, will provide exemplary research training to predoctoral students interested in the broad research areas of obesity, diabetes, metabolism, digestive diseases, endocrinology, genomics and gene therapy, epidemiology, and diseases of the kidney and pancreas. Program administration and trainee supervision will be the responsibility of the Program Director and a Steering Committee, which will meet every 6 months to review and discuss trainee progress and program enrichment. We sincerely believe that the proposed Training Program is a true investment in both the scientific future of the trainees and the public health of this nation.