PI: Ackerman, Frank
Title: Economics for Health and Environment
Abstract: During the next two years, we plan to build on our existing work and move forward in five principal areas: (1) Developing the economic theory ofprecaution. The economic case for precaution needs to be made in the realm of economic theory, as well as in more empirical terms. Conventional approaches to economic theory are only capable of analyzing environmental risks if the probabilities and magnitudes of possible harms are known in detail. Such information is typically not available in problems that require precautionary action. However, a little-known branch of economic theory rigorously analyzes decision-making under extreme uncertainty, and proves that in many cases, decisions should be based on evaluating the worst-case credible outcome — exactly the point of the precautionary principle. A brief application of this theory to dioxin and polyvinyl chloride (PVC) can be seen in Frank Ackerman’s paper presented at the Dioxin 2003 conference, which is attached to this proposal. We will continue to develop this theory, presenting it in articles for publication in economics journals, in applications to major policy problems, and in popular accounts of the precautionary principle.  (2) Developing general educational materials. The notion that protecting the environment and human health is expensive, or at odds with economic well-being, pervades public debate. In the absence of strong rebuttals to the anti-environmental rhetoric of free-marketeers, even many environmentalists believe that the goals they work for exist in tension with the goal of economic prosperity. There is an urgent need for credible, accessible educational materials to dispel this myth and make clear the reasons why it is, in fact, possible to “prosper with precaution.”  The need for practical information about the economic benefits of environmental protection is suggested by the runaway success of our “Prospering with Precaution” paper (a copy is attached to this proposal): with minimal publicity effort on our part, it has showed up on websites across the country, at major environmental conferences in Europe, and elsewhere. We plan to build on this success by expanding our analysis and distributing it through more formal channels. We plan to expand “Prospering with Precaution” for publication in an academic journal. In addition, we will explore opportunities for related publications and presentations, which will complement the labor focus of “Prospering with Precaution” with studies of the impacts of environmental protection on communities, tax revenues, and other areas of concern. (3) Responding to the abuse of cost-benefit analysis in regulatory policy. In the hands of White House “regulatory czar” John Graham and others, cost-benefit analysis has been used repeatedly
• What individuals or institutions bear financial costs associated with the transition to safer alternatives? What individuals or institutions gain from the switch?
• What are the benefits of switching to safer alternatives? In what units (avoided deaths, diseases, learning disabilities, etc.) are they naturally measured?
• And finally, how certain is our knowledge of the benefits? Is research under way that will improve that knowledge? What are the advantages or dangers of delaying action until better information is available?
(4) Analyzing chemicals policy and North-North trade. The European Union, which serves as the market for more than one-fifth of all US exports, has high and rising environmental standards. Some of the most successful American export sectors will be affected by the EU directive on electrical and electronic waste, requiring the phase-out of several chemical hazards in all electronics sold in Europe as of 2004, and REACH, the new EU chemicals policy, requiring a precautionary approach to chemical hazards. To maintain and expand these export sectors, US business needs to look forward toward meeting the precautionary standards set in Brussels, not backward toward lobbying for deregulation in Washington.  We will look in particular at the new EU electronics and chemicals policies — including economic analysis of the benefits of the REACH program, to counter vigorous industry claims of its excessive costs. In this work we will extend and deepen themes that have been broached in initial work on REACH sponsored by the World Wildlife Fund and the European Environmental Bureau. We will work proactively to develop responses to the challenges that have been or are likely to be directed against the policy by industry coalitions that see the new policy as a threat. Our critique of the abuses of cost-benefit analysis in US policy debates should provide a basis for a similar critique in the EU debate.  We will also create comparable databases of environmental impacts of production by detailed industry sector, for the US and its major trading partners. These databases will enable us to make quantitative estimates of net global environmental gains or losses when a good is exported from the US, rather than being produced in the consuming nation. Kevin Gallagher’s work at GDAE has developed similar data for US-Mexico trade; we will extend the analysis to explore the environmental impacts of North-North trade, an area that has received too little attention in discussions of globalization. In a closely related study, recently proposed to the Japan Foundation, we will examine the pollution and other environmental impacts of US-Japan trade, jointly with a group of Japanese colleagues led by Dr. Masanobu Ishikawa of Kobe University.

PI: Ackerman, Frank
Title: French Industry and Sustainable Chemistry: The Benefits of Clean Development
Abstract: In this proposal the Global Development and Environment Institute (GDAE) at Tufts University describes our plan to write a report on the future of chemicals manufacture and management in France. The report will make the case that safer products and cleaner production processes would strengthen rather than weaken the competitiveness of the French chemicals industry. It will argue that REACH and other EU regulations will provide significant benefits at minimal cost to the French economy, and will help to refute common claims about the need for elaborate risk analyses to prioritize environmental decision making.
The report will be designed to be useful in the weeks preceding the October reading of REACH in Strasbourg, but will also serve as a resource for chemicals policy planning beyond the REACH negotiations. We anticipate public release of the report in France in early October.

PI: Ackerman, Frank
Title: Safer Chemicals Management: An Economic Development Perspective
Abstract: Exposure to toxic chemicals can cause severe and sometimes irreversible damage to human health, including cancers, endocrine and neurological disorders, birth defects, and learning disabilities. In addition to the human suffering produced by avoidable toxic exposures, these exposures have economic effects. The costs of treating environmentally induced illnesses deplete the resources of individual families and can strain the capacity of government agencies that provide health and social services. When fetal, infant, or childhood exposure to toxic chemicals impairs the ability of children to learn and develop normally, the long-term costs to an economy can be enormous. The economic problems associated with toxicexposures have been explored in more detail in the context of the developed world, but the problems are often even more severe in developing countries.

The public health effects of chemical exposures, and the attendant economic impacts, make it particularly important for economic development plans to include a chemicals management component. In the proposed study, we will make the case that better chemicals management and stronger chemicals control can have significant economic benefits and contribute to economic growth, social welfare and poverty reduction. The study will review and interpret results from existing case studies that have been carried out in the developed as well as the developing world. Our study will draw an explicit link between chemicals management decisions and the overarching goal of poverty reduction.

PI: Agyeman, Julian
Title: Urban Audiences and Environmental Education
Abstract: The central issue of MassAudubon’s RFP, namely effectively engaging diverse audiences in environmental and conservation-based education whether sanctuary or classroom based, has been a critical agenda item for many years for organizations such as the North American Association for Environmental Education (NAAEE) and World Wildlife Fund (WWF). The same can be said for engaging diverse audiences in environmental careers, as pioneered by the Environmental Careers Organization (ECO). National and international organizations such as these, together with countless local initiatives, academics, practitioners and consultants have researched and developed ideas which represent parts of the jigsaw, but there is seemingly no ‘answer’, no magic bullet. Instead of universal solutions, it seems more likely that there are contingent and context specific approaches that may work in the case that they were designed for but not in others. For this reason, we commend MassAudubon for investigating the circumstances at play in their two study sanctuaries, and not just looking for formulaic or ‘off the shelf’ answers.  Here’s a selection of what we do know however. First, the word ‘diversity means many things to many people. While we have a view as to what it means, this may not be MassAudubon’s view. We need to clarify whether in MassAudubon’s view this means all of racial, socioeconomic, cultural, religious, sexual, gender, ability/disability, age or a more focused selection. Second, African Americans as Mohai (2004)1 has shown do place a high value on environmental sustainability. They are as concerned about global environmental and nature preservation issues as whites, but are more concerned about pollution issues especially in local neighborhoods than whites. Third, a conservation workforce of limited diversity is a problem for two reasons. The first is that it raises questions of legitimacy. If people of color are not represented as educators at urban sanctuaries, communities of color are less likely to perceive the facility as a broadly public endeavor representing the needs and interests of the entire community. This is extremely injurious to the success of conservation work. The second is that a workforce that is not diverse cannot expect to arrive at effective solutions that arise from different perspectives and experiences that will increase audience diversity. Fourth, research in the UK (Agyeman 1995, 1998) has shown that interest in ‘multicultural’ urban nature, that is non-native plants from around the world which exist in our cities, can be a good introduction, a ‘stepping stone’ to the more specialized concepts inherent in ‘conservation’ for diverse urban audiences.

PI: Baleja, James
Title: Molecular Interactions in Signaling and Vesicle Sorting
Abstract: Proteins containing EH domains and proteins containing Asn-Pro-Phe (NPF) sequences are ubiquitously involved in fundamental molecular sorting mechanisms. Reps1, an EH domain-containing protein, and FIP2, an NPF-containing protein, are downstream targets of Ras and Ral GTPases and the EGF receptor, suggesting that they are involved in both signal transduction and vesicle-mediated trafficking. Despite the importance of the EH domain:NPF interaction in building the protein networks responsible for vesicle-mediated transport and signal transduction, the factors that regulate their association are largely unknown. Knowing the basic principles that govern their regulation will lead to strategies to control these proteins and a better understanding of their biological roles. The EH domain of Reps1 binds directly to the NPF sequence of FIP2. Using Reps1:FIP2 as model system, the objective of this proposal is to evaluate the factors that modulate EH domain: NPF interaction. EH domains bind Ca2+ using the EF-hand fold. In Aim 1, EH domain affinity for Ca2+ and Mg2+ will be measured to test a new regulatory hypothesis operating in vesicle sorting. To examine the conformational changes that accompany metal ion binding, the structures of the apo and Mg2+-bound domains will be determined using NMR and compared to the Ca2+-bound structure. FIP2 and Reps1 both oligomerize through a coiled-coil domain. In Aim 2, we will determine the structure of the FIP2 coiled-coil domain and how oligomerization governs the strength of the interaction between these proteins. To understand the mechanism of binding, the structures from a series of unbound peptides as well as an EH domain-NPF complex will be determined in Aim 3. An inhibitory peptide, designed to be stable in cell-based assays, will also be optimized and tested as a tool to understand the roles of EH domains in vesicle-mediated internalization and secretion. Ultimately, the results of these experiments will provide insight into the vesicle sorting mechanisms important for diverse processes such as fertilization, neurotransmission, and cell growth. These investigations on the EH domain: NPF interaction are also expected to advance the application of NMR in understanding molecular recognition.

PI: Beinfeld, Margery
Title: CCK in Generating Enzyme: Structure, Location and Role
Abstract: Prohormone convertases PC1, PC2 and PC5 are good candidates for the enzymes responsible for the post-translational processing of CCK in rodent brain. PC2 is essential for normal pro CCK processing in some mouse brain regions, although there is clearly another enzyme(s) that processes pro CCK in place of PC2. Knowing the extent of colocalization of PC1 and PC5 with CCK will indicate whether they are candidates for the enzyme(s) that work in concert with PC2. We have made significant progress toward understanding how the sequence of pro CCK determines where it is cleaved and what cleavages are required for production of amidated CCK in an endocrine cell line. Proposed site-directed mutagenesis and expression studies would extend our understanding of this process. Tumor cells have been very useful to study pro CCK processing, but they are not good models for pro CCK processing in rodent brain. The use of antisense strategies to inhibit the expression of these enzymes in organotypic rat brain slices will allow us to learn more about their role in CCK processing. Together these experiments represent an integrated and novel approach to understanding the biosynthesis and processing of pro CCK in endocrine tumor cells and in rodent brain. The experiments test the following hypotheses: 1) PC1, PC2 and PC5 comprise a redundant system to insure production of active CCK in rodent brain. 2) Pro CCK has 3 dimensional structure that influences where it is cleaved. This structure changes as it is processed, making other sites more accessible to subsequent cleavage. The overall goal of this work is to understand the mechanism and enzymology of post-translational processing of pro CCK. CCK is known to be an important element in the neurochemical balance that is essential for normal nervous system function. The following specific aims are proposed: 1. The distribution and co-localization of PCI, PC2 and PC5 mRNA with CCK mRNA will be determined using in situ hybridization histochemistry in rat brain. In parallel, the colocalization of PCI, PC2 and PC5 enzyme protein with CCK immunoreactivity will be determined using immunohistochemistry. 2. Ongoing studies on the effect of altering the sequence of rat pro CCK on its processing in pituitary At-T20 cells will be continued. 3. The importance of PCI, PC2 and PC5 for pro CCK processing will be determined using organotypic rat brain slices in culture. Inhibition of enzyme expression will be achieved by incubation with peptide nucleic acid compounds, specific antisense oligonucleotides, double stranded RNA, DNA transfection and/or treatment with viral vectors expressing antisense cDNAs. The effect of this inhibition on the processing of pro CCK will be determined.

PI: Bers, Marina
Title: CAREER: Virtual Communities of Learning and Care: Multi-user Virtual Environments that Promote Positive Youth Development
Abstract: We live in a society where concepts of self and community are constantly changing. This context makes it challenging for young people to construct a sense of self and grow into contributors to civil society. This becomes even more challenging for youth suffering from a severe chronic illness that requires medical interventions such as heart and kidney transplant. In the past, most of this young people wouldn.t survive. Today, the current advances in medicine make it possible not only to extend their length of life, but also their quality. However, major life style changes and compliance to demanding medication and dietary regiments are needed. Technology can tackle this challenge by integrating already existing face-to-face psychoeducational programs with virtual environment aimed at fostering community building and social support. My career goal is to develop a groundbreaking research program and teaching career in the area of technologies to foster youth positive and healthy development. My proposed first step, through this career award, is to work at Boston's Children's Hospital with young people suffering from severe renal and cardiac failure that wouldn't survive without an organ transplantand who are at risk of developing mental-health related problems. Later on, I plan to extend this work to other settings for young people at risk, such as community-based organizations and after-school programs. I propose to investigate how multi-user virtual environments specifically designed to foster new kinds of communities of learning and care can lead to positive youth development. I coined the term identity construction environments (ICE) to refer to these technologies. The applied developmental science model provides a framework to design ICE. It also provides a model for doing research in complex real-world settings. It is my career goal to build on this framework by exploring three general hypotheses. First, ICE promote positive youth development, measured in regard to the development of the six C's of positive development: competence (i.e. compliance and technological fluency), connection, character, confidence, caring and contribution to civil society. Second, ICE complement and augment the effects of face-to-face psychosocial interventions. Third, the positive effects are due to the design features of ICE and the nature of the on-line activities that engages youth in cognitive, social and emotional development. Exploring these hypotheses involves conducting interdisciplinary research, developing new technologies, such as the Zora graphical multi-user environment, and a set of assessment and professional training strategies, and critically evaluating them in the complexities of a real-world setting, such as Boston's Children's Hospital. This three-fold program makes 1) theoretical contributions by providing an interdisciplinary framework to enable collaboration between computer scientists and child development experts; 2) innovative designs of technologies that can be scalable and sustainable to promote positive youth development; and 3) empirical research to evaluate results of interventions using these technologies in real-world settings. As part of this career plan, educational activities will involve research opportunities for undergraduate and graduate students in an interdisciplinary research group; "service learning" in health care and community-based settings and new courses and curricular materials for students from a wide variety of disciplines: education, mental health, engineering and computer science.

PI: Blumsack, Marilyn
Title: Bernard Osher Foundation TILL Funding
Abstract:  Founded in Fall 2000, The Tufts Institute for Lifelong Learning (TILL) is now a mature, proven center of life long learning for the retired and near-retired population in the greater Boston area. During calendar year 2004, it served 23 participants who accounted for 435 course enrollments. It offers study groups of varying length, a Lunch and Learn series, and regular off-campus special events. It maintains a collaborative relationship with Lexington’s Brookhaven retirement community.  TILL seeks support from The Bernard Osher Foundation for two purposes: to increase its membership and to broaden the scope and number of its core eight-session and four-session study group offerings. These objectives are not mutually exclusive. Both are essential to achieving the financial strength that will ensure the Institute’s future.

PI: Boghosian, Bruce
Title: Entropic Lattice Boltzmann Models and Quantum Computation
Abstract:  Type II quantum lattice-gas algorithms have been proposed for many applications in physical modeling. Certain of these algorithms have already been implemented on NMR quantum computers. In the course of AFOSR grant F49620-O1-1-0385, we demonstrated that all Type II quantum lattice-gas algorithms are equivalent to a certain subset of classical lattice Boltzmann models. In spite of these recent advances in understanding, several interesting and fundamental issues remain outstanding. First, though we may now associate a classical lattice Boltzmann model to any Type II quantum algorithm, it is not known if the reverse is true. Of particular interest in this regard are the recently discovered entropic lattice Boltzmann models — also a focus of work in AFOSR grant F49620-O1-1-0385. Such models are capable of a smooth transition between conservative and dissipative behavior, and there are numerous reasons to believe that they may have Type II quantum algorithms as counterparts. Finally, no unified complexity model for Type II quantum computers has yet been elucidated. It is likely that such a model may be formulated in terms of the density matrix describing both the quantum and classical degrees of freedom present in the Type II computer.

PI: Bratt, Rachel
Title: Women's Institute for Housing and Economic Development: 1981 – 2006
Abstract:  The mission of the Women’s Institute is to build affordable housing that fosters economic security for low-income women and families. For nearly 25 years, they have worked with community-based organizations to create affordable housing and programs for low-income and homeless women and families.

Recognizing that families benefit from community-based programs and thrive in environments where they can develop peer supports, WIRED has have worked in partnership with over 50 community-based organizations that provide programs and activities to increase women’s economic and family stability. Their more than 420 completed units of innovative housing include community spaces that promote skill development and family well-being. In short, WIHED has created programs that give women the tools to make sound economic choices for their families.  Building on their history and expertise, WIRED currently has over 100 units of community-supported affordable housing in production and many more in the pipeline.  As part of their efforts to increase their internal capacity, and in order to deepen and extend their impact in Boston and nearby communities, WIHED has indicated an interest in partnering with the Department of Urban and Environment Policy and Planning (UEP). The UEP faculty has longstanding relationships with Boston-area nonprofit organizations and a commitment to community-based development. UEP also combines a unique focus on housing, community development, and child and family issues.  This contract will cover the costs involved in developing: 1) a case study of the organization and 2) an evaluation plan to assess current and future projects.

PI: Brenner, Brian
Title: Developing a Library of Bridge Models
Abstract:  This proposal is an extension of work by the Tufts University Civil and Environmental Engineering Department, working with fellows of the NSF funded Tufts Engineering the Next Steps (TENS) GK-12 Project, to develop buildable model bridges of different structural types. The models are used for outreach in Massachusetts classrooms, grades 1-6. The working models demonstrate different types of bridge design, architecture, and construction methods as they are assembled by grade school students in a way that simulates the actual construction process. This library of bridge models will be used to foster outreach to schools, interest in K-12 education by current college students, and awareness of architecture and design in our communities.  Outreach to public K-12 schools by members of the industrial and academic communities is a vital step in developing and fostering life-long learning in classrooms across the nation. By bringing the experience and content expertise of those in industry or academics into the K-12 classroom setting, students receive up to date information from “professionals” that engage their interest and possibly inspire them to pursue careers in the fields with which they are now familiar The development of a bridge model library provides opportunity for young, college level engineers to enter the K-12 environment to act as role models for younger students while teaching them about bridge design and the importance such work has in society. The process of model building in the classroom includes clear benefits to younger students, but also benefits to college students who design the models, educational presentation boards and materials, and who lead the presentation and class room discussion. Presenting to a 4th grade classroom is not unlike presenting to a marketing team or others groups in industry that have no real technical training. Thus, the communication skills of college level engineers are strengthened such that these students become more marketable post graduation.  This grant will supplement overall efforts of the Tufts University Center for Engineering Educational Outreach (CEEO). The CEEO has been developing tools, methods, and support systems for integrating engineering into K-12 schools for over 15 years. Teaching through engineering fosters a hands-on learning experience that is effective with all types of learning styles. The CEEO has expertise and experience in conducting outreach in public school districts and is therefore suited to bring the proposed bridge models to classrooms for presentations across Massachusetts. 

PI: Bridges, Robert
Title: Research Training For Tufts Veterinary Students
Abstract: There is a need for veterinarians with research training to participate in academic as well as corporate based research in the fields of animal and human health. It is crucial for a set of 21st century veterinarians to acquire the scientific skills and technical training together with the conceptual framework to participate both as independent researchers and collaborators to meet the projected research needs in biomedical sciences. Through the training set forth in the present proposal, the objective is to provide bright, and highly motivated veterinary students with a one-year, in-depth research experience in a productive and active research setting. Training will involve the use of animal models to develop skills at hypothesis-based, biomedical research. Over a three-year period, the aim is to train a minimum of nine students (three per year). Applicants will be actively recruited from the incoming and existing veterinary classes, with an effort made to insure participation by our minority students. Students will receive research training during a consecutive 12-month period after their first year of veterinary school, but prior to graduation. A program faculty that consists of 20 faculty from the three departments with expertise in numerous research areas that use animal models will offer both intensive laboratory training as well as complementary seminars and course work. Key research areas include infectious diseases, reproductive biology, biotechnology, digestive diseases, neuroscience and behavior, oncology, nutrition, and respiratory physiology. A wide variety of animal models are available for trainee projects, including pigs, horses, goats, sheep, cats, dogs, rats, mice, and shrimp. Highly successful projects can be submitted to meet the requirements for a Masters Degree in Comparative Biomedical Sciences. All participating students will receive a certificate at commencement acknowledging their training experience. The program itself will contribute funds that can be used to support additional qualified applicants and/or supplement existing stipends. It is a long-term objective to make this training experience a launching pad for career involvement in biomedical research for the veterinary students. The success of the program will be evaluated by determining the number of trainees that integrate health science research into their professional careers.

PI: Brodeur, Peter
Title: Organization and Expression of V Genes
Abstract: Antigen specific receptors of B and T lineage cells require the somatic assembly of V, D, and J gene segments during lymphocyte development. Immunoglobulin and T cell receptor loci are targeted by a common V(D)J recombinase in a cell type- and developmental stage-specific manner. The fundamental control of this process occurs at the level of alterations in chromatin structure that confer locus "accessibility" to the RAG1/2 complex. The immunoglobulin heavy chain locus (Igh) is the first to rearrange during B cell differentiation and accessibility at different stages is limited to discrete domains. The cis-acting elements and signaling pathways that regulate accessibility throughout the lgh locus are yet to be fully elucidated. Since V(D)J recombination is required for normal lymphocyte development, defects in the process result in both subtle and catastrophic immune deficiencies. Thus, understanding the underlying mechanisms has obvious implications to health and disease. Our long term goals are focused on the control of Igh locus accessibility within the nearly 3 megabase region containing approximately 200 Vh gene segments. In this application, we propose to extend our studies of a candidate control region in the 5' boundary of the lgh locus that exhibits early B cell specific chromatin remodeling. This novel region interacts, both in vitro and in vivo, with factors that have been implicated in Igh accessibility and rearrangement. We propose to examine sorted early B cell subsets from bone marrow to determine the precise stage of B cell development at which this element is active. Gel shift and chromatin immunoprecipitation approaches will be used to define the factors that are recruited to the multiple sites mapped within this region. The function of this putative regulatory element will be tested by both genomic deletion and studies of BAC transgenes containing putative control regions and Vh segments of the 5' region of the locus. We will use a novel cell line model as well as primary bone marrow cells to determine the mechanism through which IL-7 receptor signaling selectively mediates D-distal (5') Vh gene accessibility.

PI: Brodley, Carla
Title: Automatic Detection of Covert Channels in Networks Proving the Quality of Sensor Data Through Data Mining
Abstract:  A covert channel is a mechanism that can be used to violate a security policy by allowing information to leak to an unauthorized process. Two types of covert channels exist: storage and timing channels. A storage channel “involves the direct or indirect writing of a storage location by one process and the direct or indirect reading of the storage location by another process”. A timing channel involves a sender process that “signals information to another by modulating its own use of system resources (e.g., CPU time) in such a way that this manipulation affects the real response time observed by the second process”. This classification can be taken further by identifying hybrid channels in which the timing and storage information are used together, and counting channels in which the number of events come into play instead of the occurrence of a single event. In this proposal, we focus on the analysis and detection of covert timing channels in the TCP/IP protocol suite. Although some work has been done on timing channel analysis in general, little attention has been paid to channels in IP. In the proposed work we will explore how such channels can be detected and/or rate limited.  To facilitate our research we have designed and implemented a covert channel between Tufts and Georgetown Universities. Many implementation issues were non-trivial, and we now have robust implementation that handles synchronization issues and network jitter. We are in the process of implementing some basic error correction to make the channel more robust.

PI: Camilli, Andrew
Title: Short-Term Training for Minority Students Program
Abstract: The Sackler School of Graduate Biomedical Sciences of Tufts University seeks renewal of NHLBI funding for a Summer Research Internship Program designed to provide research opportunities to underrepresented minority students, and in this way, also address the shortage of qualified minority investigators in biomedical research. The Sackler program has been in operation for twelve years and has trained 160 minority students during this period. The objectives of this program are: 1. To increase participation of minority undergraduates in biomedical research through summer research internships at the Tufts Health Sciences Campus. 2. To enrich this internship experience with training in communication skills, career counseling, and networking activities. 3. To involve more faculty, postdoctoral fellows and graduate students on the Health Sciences Campus in the mentoring of minority students. 4. To increase the awareness of minority students as to the benefits of biomedical research and potential careers in this area, particularly in the areas of cardiovascular, pulmonary, hematologic and sleep disorder research. This program, consisting of a ten-week summer research internship for 18 trainees, will give the students an opportunity to interact with, and work alongside outstanding research scientists. It will help them decide if biomedical research is a career they wish to pursue. For those who have already decided on biomedically oriented careers, the training will give them the confidence and additional qualifications to apply to, and succeed in, the best programs. Our long-range goal is for these students to become independent scientists in academia or industry, and in this capacity, serve as mentors for other minority students.

PI: Camilli, Andrew
Title: Study of Transmissible Forms of Vibrio Cholerae
Abstract: Vibrio cholerae, which lives in association with plankton in brackish, temperate waters the world over, is the causative agent of endemic and epidemic cholera. Hallmarks of the disease include prodigious watery diarrhea resulting from the action of secreted cholera toxin (CT), and infrequent but deadly explosive epidemics. The strong link between explosive epidemics and human crowding accompanied with untreated drinking water suggests a very efficient mode of fecal-oral transmission. We have discovered a heightened state of transmissibility of stool V. cholerae (referred to simply as "hyperinfectivity"), which persists even after shedding into water reservoirs. Knowledge of the molecular basis for this phenotype, and a general characterization of this transmissible form of V. cholerae, would contribute to the design of vaccines to prevent cholera at the initial stage of infection. Aim 1 of this proposal will use transcriptional profiling and proteomics to help define this transmissible form. Spotted DNA microarrays will be used to determine the transcriptome of stool V. cholerae incubated in pond water, and this will be compared to that of fresh stool V. cholerae to identify potential differences. The results will be validated by quantitatively assaying the steady state mRNA and protein levels from select genes. Microscopy and transcriptome data on stool V. cholerae predict a bacterial state of motility working in the absence of chemotactic signaling. This counterintuitive state is hypothesized to be responsible, at least in part, for the hyperinfective phenotype. In Aim 2 of this proposal, quantitative immunodetection using paralog-specific antisera will be used to test for reduced expression of all three CheW linker proteins and all three CheR methytransferases in fresh and pond water-incubated stool V. cholerae, as is predicted by current transcriptome data. In addition, capillary tube chemotaxis assays will be performed directly on V. cholerae from these samples to substantiate this hypothesis. Aim 2 will also test a second hypothesis, that ToxR regulated factors, which are essential for pathogenesis, are not playing a role in the hyperinfectious state. Finally, Aim 3 will use mutation and infectivity analyses to determine if other metabolic, physiologic or phenotypic properties of the bacteria contribute to the hyperinfective phenotype or, alternatively, to colonization of an environmental planktonic host, Anabaena variabilis. These studies will establish a basis for understanding the hyperinfective phenotype, and the properties in general, that are exhibited by fresh and pond water-incubated stool V. cholerae. In turn, this knowledge will enhance our understanding of transmission of this and perhaps other water-borne pathogens, it will aid in the development of new cholera vaccines that target the antigens of 'incoming' vibrios, and it may suggest new approaches for the prevention of the dissemination of this lethal organism.

PI: Cao, Caroline
Title: CAREER: Adapting To Technology In Minimally Invasive Surgery
Abstract: This proposal aims to combine research activity and education in an integrated multi-disciplinary program. It fits well with the goals of Tufts University in providing cross-disciplinary education and research opportunities for students in Arts & Science, Engineering, and Health Sciences.

The specific objectives of this application are the following:

Research:
1) Study the effects of evolving endoscopic technology on the surgical process in minimally invasive surgery, and how novel technology (e.g., robotics) can be used to better augment surgical performance;
2) Develop a methodology for analysing complex surgical processes in MIS, and a metric for assessing technology and surgeon performance in MIS;
3) Reduce the possibility for errors by facilitating the integration of technology in MIS and the design of effective interfaces and informational aids to enhance surgeons performance;
4) Support the training of surgeons in MIS with modular computer-based training tools;
5) Contribute to the theoretical understanding of human performance and error remediation in dynamic remote environments.

Education:
6) Develop and promote the interdisciplinary approach to complex systems in graduate and undergraduate curriculum, using the proposed research activities as course material for realistic case-based problem-solving exercises;
7) Develop and promote hands-on learning through laboratory projects and research experiences in undergraduate education, working on small projects related to the proposed research activities;
8) Develop and train successful research scientists and human factors engineers through teaching and mentoring activities.

PI: Castaneda-Sceppa, Carmen
Title: Strong Living Program: A Community-Based Exercise Program
Abstract:  This is a contract for personal services between the State of Vermont, Department of Health (hereinafter called “State”), and Tufts University (Strong Living Program), with a principal place of business in Boston, MA (hereinafter called "Contractor”). Contractor’s form of business organization is corporation. Contractor is not required by law to have a Business Account Number from the Vermont Department of Taxes. The subject mailer of this contract is personal services generally on the subject of conducting training for a physical activity program for elders.

PI: Castellot, John
Title: Training Program in Developmental Biology
Abstract: Continuing support is requested for five predoctoral graduate students per annum to participate in a Training Program in Developmental Biology. Graduate students in this Program become expert in the concepts of modern cell, molecular and developmental biology and the application of molecular, cellular, morphological, and organismic methodologies to the mechanisms underlying embryonic development, reproduction, tissue remodeling, and disease processes involving alterations in cell behavior. Several major themes of research are available to the students: 1. early vertebrate development and reproductive biology; 2. tissue and organ development and remodeling; 3. disease processes, such as cancer, aging and vascular disease, resulting from disturbances in cell behavior and tissue remodeling; 4. basic molecular and cellular mechanisms relevant to development. The unique strengths of the Program derive from the established strength of the faculty, the sustained retention of a strong pool of matriculants into the Program over the past decade or more, the intense concentration and collaboration on mechanisms of dynamic cell behavior in the above systems, the broad range of technical expertise of the faculty, the availability of top-grade facilities that allow sophisticated technology to be used, the location of the Program in a very strong Health Science research environment that facilitates collaborations, the interactive nature of the Program members, and the close attention given to trainees in the small to moderate-size laboratories of the Program. On graduation, trainees will receive a Ph.D. in Cell, Molecular and Developmental Biology from the Tufts University Sackler School of Graduate Biomedical Science.

PI: Cebe, Peggy
Title: Polymer-Based Nanocomposites: An Opportunity for Deaf and Hearing-Impaired Students
Abstract: The objective of this proposal is to provide an opportunity for deaf and hard of hearing students, which involves participation in classroom and laboratory research work in the field of polymers.  A small group of  college-age students will perform an internship  at  Tufts  University  for  six  weeks  during  the  summer.    The  educational opportunity is arranged in two parts, the classroom component and the laboratory component.  Sign language interpreters will be provided for all the classroom lessons and the group laboratory instrument training sessions.  A field trip to the National Plastics Center and Museum will be undertaken. The  classroom  component  will  address:  1.  basic  polymer  science  needed  to conduct the laboratory research work, and 2. aspects of participating and communicating within  a  scientific  environment.    From  the  polymer  science  lessons,  students  will  learn about the chemistry and physics of polymer molecules, crystallization and melting of polymers, the interaction of X-rays and light with polymers, mechanical properties of polymers,  and  the  connection  between  thermal  processing,  structure,  and  ultimate properties of polymers.  To prepare the students for participation in the scientific community, a strong component of pre-professional training will be incorporated.  This training will include discussion of situational ethics pertaining to proper conduct in the laboratory (integrity in the performance of research) and ethics of writing and presenting results (integrity in scholarship).   Deaf and hard of hearing students face obstacles in communicating  with the hearing world.  A major portion of the classroom component will center on teaching the students how to communicate, including how to write a scientific report, how to maintain a laboratory notebook, how to make a group meeting presentation, and how to organize a poster presentation.  The students will present their results at a weekly  group meeting, and will write a summary report at the end of the internship.  In  the  laboratory  component,  the  students  will  be  making  and  characterizing polymer-based nanocomposite films.  In the first year, the nanocomposites will comprise a  semicrystalline  polymer  matrix  (eg., poly(vinylidene  fluoride))  and  an  organically modified  silicate  (OMS)  filler  (eg., NanomerTM,  a  modified  montmorillonite  clay).  Composite  films  will  be  cast  from  solution,  thermally  treated  to  affect  microstructure development, and then  characterized using techniques such as: differential scanning calorimetry (DSC), wide angle X-ray scattering (WAXS), polarizing optical microscopy (POM), and Fourier transform infrared spectroscopy (FTIR).  Mechanical properties will be assessed using tensile testing.  The goals of the laboratory research component are to: expose the students to the laboratory  environment; introduce them to the concepts of formulating  and  testing  hypotheses;  assist  students  to  conduct  systematic  studies  while controlling variables; illustrate the use of modern analytical equipment; and demonstrate the connection between processing variables, structure, and properties. In interdisciplinary team of professors from Physics (Peggy Cebe, Bob Guertin), Chemistry (Terry Haas), and Biological and Chemical Engineering (Regina Valluzzi) are volunteering their time to assist with this program.  A graduate laboratory assistant will help with the laboratory and classroom components, preparation of educational materials, and development of a web site for dissemination of results.

PI: Chapra, Steven
Title: Improved Science and Decision Support for Managing Watershed Nutrient Loads
Abstract:  This proposal focuses on two major gaps in that presently impede the effective management of nutrients in watersheds: (1) inadequate characterization of sediment-water interactions and fixed plant activity in current receiving water quality models and (2) lack of a decision oriented framework for managing nutrient loads in a watershed. A decision support system (DSS) will be developed which integrates scientific models of watershed nutrient loads and their fate in receiving waters, into a decision-oriented optimization framework. The DSS model development process will allow stakeholders, scientists and regulators to define and prioritize watershed nutrient management objectives at the early stages of the project. This process will lead to an initial screening level DSS by the end of year one, based on simple nutrient loading function models, GIS information, a simple receiving water model and a decision model driven by a genetic algorithm. The initial DSS will be tested and evaluated by local scientists, stakeholders and regulators and used to formulate a more realistic DSS in subsequent project years, by incorporating more sophisticated runoff and receiving water models into the framework. The proposed DSS will be used to develop a TMDL program and to design a water quality monitoring network for the Mystic River Watershed. The Mystic River watershed is attractive because it is made up of both agricultural and urban land-uses and consists of a network of streams and lakes. In addition, the river is an impaired waterway and contains a current USGS NAWQA station. An existing collaborative partnership and close relationship already exists between the research project team, Tufts University and the Mystic River Watershed Association.

PI: Coffin, John
Title: Molecular Biology of Retroviruses
Abstract:  We will determine the nature of viral and cellular factors which affect the specificity of integration for specific sites and regions of target DNA. We will also develop arid exploit biochemical and genetic approaches to examine the interaction between IN and its viral DNA substrate. We will attempt to reconstruct preintegration structures that carry out the reaction correctly in vitro and when introduced into cells.

PI: Coffin, John
Title: Retrovirus Evolution
Abstract: Retroviruses exhibit a wealth of evolutionary phenomena, including the ability to undergo rapid genetic change in response to varying selective pressure; the ability to vary in the use of host cell receptors; and the ability to become integrated in the genome of their host species and passed down through the generations as endogenous proviruses. In the prior project period, we have studied all these aspects of retrovirus evolution: We have looked at the mechanism of evolution of env genes by analyzing an unusual mutant that extends the host range of ALV beyond chicken, to quail, dog, and even human cells. We have isolated and studied an unusual mouse endogenous provirus that appears to occupy a special phylogenetic location between mouse and non mouse species. We have extensively analyzed the coevolution of humans and their endogenous proviruses, particulary the relatively recently inserted HERV-K family. We have developed sophisticated mathematical models for the evolution of replicating virus populations, describing the effects of mutation, selection, drift, linkage, and recombination on the accumulation (or loss) of deleterious mutations. Future work will address the following aims: 1. How do retroviral envelope genes evolve to use new receptors and to alter other important properties? We will use our extended host range mutants to test specific hypotheses for this process. 2. What are the functional and pathogenic properties of human endogenous retroviruses, particularly HERV-K? Can we isolate infectious virus? Is their expression or reintegration associated with malignancy? 3. How do important the forces of mutation, selection, recombination, and drift combine to direct retrovirus evolution? We will combine mathematical and in vitro modeling of virus replication to test specific evolutionary models.

PI: Dawson, Dean
Title: The Meiotic Role of Slk19p in Yeast
Abstract: A fundamental difference between meiotic and mitotic chromosome segregation is that in meiosis I, sister chromatids move as a unit to one pole of the spindle rather than separating as they do in mitosis. Sustained linkage of sister chromatids through meiosis I is accomplished by association of the chromatids at the centromere region and development of kinetochores that allow both chromatids to be moved as a unit to one pole of the spindle. The localization of the meiosis-specific cohesin, Rec8p, to the centromeres is essential for maintenance of sister chromatid cohesion through meiosis I, and Mam1p is necessary for kinetochore development, but the molecular basis for the regulation of cohesion and sister kinetochores in meiosis remains a mystery. Recently, it has been demonstrated that the S1k19 protein of Saccharomyces cerevisiae is essential for the maintenance of sister chromatid association through meiosis I. Two possible roles have been suggested for Slkl9p in meiosis. The first is to promote sister chromatid cohesion at the centromeres, perhaps by protecting Rec8p in this region from degradation at the metaphase I to anaphase I transition. The second is to control sister kinetochore development, such that sister chromatids share a single functional kinetochore throughout meiosis I. This proposal addresses these models through four sets of experiments. First, experiments are proposed that use the chromatin immuno-precipitation method to explore the centromeric localization of Slkl9p and Mamip, and their dependence upon each other for association with the centromere. The role of the CDEII centromere element in meiotic kinetochore function will be explored genetically and through studies of its association with Slkl9p. Second, we will test whether Slk 19p controls sister chromatid centromere cohesion by monitoring its relationship with Rec8p and by using cell biological assays to monitor the establishment and maintenance of sister chromatid centromere association in slkl9 mutants. The third set of experiments is designed to determine the identities of the proteins that interact with Slkl9p in meiosis. A two-hybrid screen will be performed. Affinity chromatography will be used to purify Slkl9p and associated proteins from meiotic cells. A genetic approach will used to identify high copy suppressors of slkl9 partial loss-of-function mutants. Finally, we will explore the regulation of Slkl9p by: 1) Spo 13p 2) possible conjugation to ubiquin or ubiquitin-like proteins, 3) targetting for degradation by Cdh 1p and Amaip and 4) degradation by Espip, the protease that triggers anaphase I by clipping cohesin proteins.

PI: Durant, John
Title: Post-Alum Treatment Testing, Testing for Pre- and Post-BMP Catch Basin Installation, and QAPP Development for Spy Pond in Arlington, MA
Abstract:  Task 1: Post-alum treatment testing and monitoring with semi-annual reports.  Following the application of the alum (in May or June 2004), monitoring will be done in the two basins of Spy Pond every 2-4 weeks to measure the changes in phosphorus levels. Monitoring will be done throughout the year, but less frequently during the winter months when productivity is low. Monitoring will be done at four locations: the pond inlet (Rte 2 storm drain), the outlet, and at fixed moorings in the deepest parts of the north and south basins. At the fixed moorings, monitoring and sampling will be done at regular intervals along a vertical transect to measure changes in water quality as a function of depth. The depths will be consistent with those used for the pre-alum treatment testing (i.e., 0.2, 3, 6, and 6.5 m in the south basin; 0.2, 4, 9, 10 m in the north basin). On each sampling trip, in situ monitoring will be done for temperature, pH, conductivity, dissolved oxygen. Flow will also be monitored at the inlet and outlet; visibility will be measured as function of depth at the fixed moorings. At all sites, water samples will be collected for laboratory analysis of total and dissolved phosphorus, dissolved aluminum, and total and dissolved arsenic. Arsenic will be monitored for two reasons: (1) there is a lot of arsenic in the pond sediments due to arsenical herbicide applications in the 1950s and 1960s; (2) the biogeochemical cycling of arsenic is similar to phosphorus, and thus it is hypothesized that the alum will be effective in preventing the remobilization of arsenic to the water column. All monitoring, sampling and chemical analyses will be performed in accordance with the QAPP “Water Quality Monitoring for the Spy Pond Restoration Project” (August 2002, revision 1).  The results of the sampling and monitoring will be reported every six months for two years following the alum application The written reports will include tables containing the field and laboratory data, estimates of pollutant load reduction brought about by alum treatment, and a discussion of field and laboratory data quality control. Pollutant load reduction will be quantitatively described.

Task 2: Quality Assurance Project Plan (QAPP) for s.319 Stormwater Management—Installation of leaching catch basins and baffle tank catch basins.  A QAPP will be developed that describes all of the field and laboratory analytical methods as well as QA/QC and reporting protocols for measuring the effectiveness of the leaching catch basins and baffle tank catch basins. A first draft of the QAPP will be submitted to the MA DEP and EPA by July of 2004 with the goal of having the QAPP approved by September 2004 so that Task 3 activities can be performed in accordance with an approved QAPP.

Task 3: Testing the current conditions before the installation of Best Management Practices (BMP) baffle tank and leaching catch basins.  The goal of this task is to assist the engineering design firm (CEI) in finding the best possible locations for the BMPs in the Spy Pond watershed. It is anticipated that 22 existing catch basins — 11 in Arlington and 11 in Belmont — will be retrofitted with either baffle tanks or leaching catch basins. A necessary first step in this task will be to meet with CEI and representatives from the Arlington and Belmont DPW’s to get detailed street plans showing the locations of existing catch basins and storm drain infrastructure. Arrangements will then be made to inspect that catch basins and determine which, based on topography, geology, and other physical constraints, are the best candidates for retrofitting. Once this list has been generated, experiments will be planned to determine which of the “best candidate” catch basins contributes the greatest amount of phosphorus to the pond.  In accordance with the RFP at least four storm events will be monitored. Samples will be collected at as many of the “best candidate” catch basins as possible. Because flow changes over the course of a storm, several samples must be collected at each catch basin to determine the range of phosphorus concentrations and loadings. When a sample is collected flow (discharge) will be measured simultaneously. The samples will be analyzed for total and dissolved phosphorus and total suspended solids (TSS). All sampling and laboratory analyses will be performed in accordance with the QAPP (Task 2).  An analytical report will be prepared based on the pre-construction water quality monitoring. The report will include quantitative estimates of phosphorus and TSS loads for each of the “best candidate” catch basins studied, and a discussion of field and laboratory quality control methods. If necessary, a meeting with CEI and the DPW’s will be arranged to discuss the Task 3 findings, and to finalize the list of catch basins in Belmont and Arlington that will be retrofitted.

Task 4. Testing post-installation of BMP baffle tank and leaching catch basins with semi-annual reports. The goal of this task is to determine the phosphorus and TSS removal efficiencies of the retrofitted catch basins. Assuming that all 22 catch basins are retrofitted as planned, it is proposed that at least eight catch basins be studied — four in Belmont and four in Arlington. Attempts will be made to study both baffle tanks and LCBs in each community. Selection of the catch basins will be made in consultation with the design firm (CEI) and the Town of Arlington. It is anticipated that removal efficiencies will be measured for each of the eight catch basins during at least three storms.  During each storm, several samples will be collected in the catch basins to determine the range of phosphorus concentrations and loadings over the duration of the storm. Flow (discharge) into the catch basin will be measured each time a sample is collected. The samples will be analyzed for total and dissolved phosphorus and total suspended solids (TSS). All sampling and laboratory analyses will be performed in accordance with the QAPP (Task 2).  Analytical reports will be prepared semi-annually starting six months after the catch basins are retrofitted. The reports will include quantitative estimates of phosphorus and TSS removals efficiencies for each of the retrofitted catch basins studied, and a discussion of field and laboratory quality control methods. Pollutant load reductions will be quantitatively described in units of mass per year and mass per unit of flow.

PI: Easterbrooks, Ann
Title: Touchpoints Early Child Care and Education Evaluation
Abstract:  Since its inception, The Brazelton Center’s Touchpoints Program (TP) has garnered much public and professional acclaim. The Touchpoints Program, built on Dr. Brazelton’s vast clinical experiences helping parents manage their young children’s often bewildering and stressful developmental transitions, trains a wide variety of child-serving professionals; its primary goals are to increase professionals’ knowledge of child, parent, and family development, and to expand professionals’ repertoire of respectful, collaborative, and strengths-oriented strategies for working with parents. Improved relationships between professionals and parents, and between parents and children, should result, and these in turn should yield enhanced child well-being.  The child development literature provides ample documentation for these basic assumptions: a) children thrive in sensitive relationships--when their caregivers (parents and child care providers) are able to “read their cues” and “interpret their signals” accurately, and respond with warm care and appropriate stimulation (Baumrind, 1991; DeWoiff & van Ijzendoorn, 1997); and b) children are influenced by the network of relationships in which child development unfolds (Cummings & Davies, 1994; Howes, 1999; Howes, & Hamilton, 1992; Owen, Ware, & Barfoot, 2000; Smith & Hubbard, 1988). In other words, positive relationships between parents, and between child care providers and parents facilitate optimal child development.

PI: Fantini, Sergio
Title: Optical Spectroscopy and Imaging of Tissues
Abstract: This project combines (1) a strong and innovative research activity aimed at the development of novel techniques for optical spectroscopy and imaging of tissues, and (2) a multidisciplinary educational plan that includes significant hands-on experience, laboratory experiments, and active learning approaches. The objective of this project is to synergistically develop the research and educational components in a mutually beneficial way. Specifically, the course material will be organized to give a solid scientific background to the students who will contribute to the research activity, while the research activity will generate a number of laboratory set-ups and sub-projects that will enhance the course teaching material and will give additional opportunities for research experience to undergraduate and graduate students. The research component of this project consists of the development of new methods and instrumentation for biomedical applications of near-infrared spectroscopy and imaging. The research field devoted to the optical study of tissues has continuously grown in the last decade leading to a number of clinical pilot studies that have recently demonstrated practical feasibility. This project aims at further advancing the field (1) by implementing and testing a new idea for improving the spatial resolution of optical tomography, (2) by developing a frequency-domain instrument operating over a large bandwidth (0.1-1 GHz) of modulation frequencies to measure tissue optical properties, and (3) by developing an instrument to measure quantitative absorption spectra of tissues over a wide wavelength range (600-1,000 nm). Our long term goal is to exploit these new developments towards novel and more powerful applications of optical spectroscopy and imaging to the biomedical field. In this perspective, one initial application (spectral characterization of tumors in vivo in an animal model) is planned during the course of this five-year project. Envisioned future applications include enhanced approaches to optical mammography, functional brain imaging, skeletal muscle oximetry, assessment of tissue hemodynamics, and providing a new technique that achieves a compromise between the highresolution/
low-penetration-depth of microscopy and the low-resolution/high-penetration-depth of photon migration in the optical imaging of tissues. The educational component of this project consists of (1) the development of a new course on the principles of medical imaging, (2) the design of new teaching laboratory experiments for two courses on medical optics, (3) the development of short research projects suitable for undergraduate summer internships, and (4) the implementation of active and collaborative learning, and of evaluation methods that include written and oral presentations that are modeled on typical job-related activities (technical manuscript preparation, grant submission, conference presentation, etc.). The proposed educational activity puts in practice two key ideals of this application, namely, the high value of multi-disciplinary education and the importance of active learning. With regards to the multi-desciplinary nature of this project, our vision is that engineers who have a good scientific knowledge of the targeted application will be most effective at producing innovative instrument designs to advance the field. Conversely, clinicians who understand the technical capabilities (and limitations) of the instrumentation they use, will be in a better position to excel. Stimulate active learning is one of the broad objectives of this project. The relevance given to hands-on activities and laboratory experiments in the proposed educational effort is one of the key approaches to achieve this objective. The multi-disciplinary plan of this proposal is coherent with the programmatic goals of the Bioengineering Center at Tufts University.

PI: Forrester, Janet
Title: Nutritional Status in HIV-positive Hispanic Drug Abusers
Abstract: The BIENESTAR study is a longitudinal cohort study of the effect of drug abuse on nutritional status and outcomes among Hispanics with HIV infection. The cohort is comprised of 3 groups: HIV+ drug abusers, HIV- drug abusers, and HIV+ non-drug abusers. Efforts will focus on expanding and maintaining this unique cohort. New hypotheses addressed in this competitive renewal reflect current trends, as HIV becomes a chronic disease. Liver disease is an increasingly common complication in patients with HIV. While it is generally accepted that HCV is largely responsible for liver disease in HIV infection, studies suggest that drug abuse itself is a risk factor for liver disease among persons with HCV. Animal studies have shown that cocaine can cause liver toxicity by the induction of oxidative stress with tissue damage. Hispanics of the NE USA preferentially inject cocaine more than other races, and therefore may be at increased risk of oxidative stress and liver dysfunction. Low serum antioxidant micronutrient levels are common in drug abusers, further increasing the risk of oxidative tissue damage. This study proposes to examine antioxidant status, oxidative stress, and liver dysfunction in Hispanic drug abusers with HIV infection. There are 4 specific aims: 1) To examine the association between the type (cocaine versus heroin) and frequency of drug abuse with plasma antioxidant capacity and oxidative stress. The specific interest is to know if cocaine differs from heroin in its ability to alter antioxidant capacity and cause oxidative stress; 2) To examine the association between the type and frequency of drug abuse and liver dysfunction. If cocaine or heroin cause oxidative stress to hepatic tissues this may result in detectable alterations in liver function; 3) To examine the association between oxidative damage and liver dysfunction. An association between oxidative stress and liver dysfunction will imply that drug abuse alters liver function through a mechanism involving oxidative stress; and 4) To examine the influence of infection with HIV, infection with HCV, and alcohol consumption on the associations described in specific aims 1 to 3. This will determine if drug abuse is an independent risk factor for liver dysfunction or whether, like alcohol, it is an accelerator of liver dysfunction in persons with HIV and/or HCV infection.

PI: Frank, Eric
Title: Development of Sensory/Motor Synapses in the Spinal Cord
Abstract: The correct operation of the nervous system depends on the formation of highly specific patterns of axonal projections and synaptic connections among nerve cells. To understand how this occurs, the approach taken in this proposal is to study the development of one well characterized set of synaptic pathways in the spinal cord, those involved in the reflex control of muscle length and tension. Sensory neurons supplying muscle spindles (Iα afferents) are primarily sensitive to muscle length; they project into the spinal cord where they make monosynaptic connections with particular subsets of motoneurons. Other proprioceptive sensory neurons (Iβ afferents), supplying Golgi tendon organs that are sensitive to active muscle tension, also make specific connections with motoneurons, but these connections are mediated polysynaptically. These issues will be addressed using the isolated neonatal mouse spinal cord preparation. A major advantage of this preparation is that different classes of proprioceptive afferents (Iα and Iβ neurons) can be stimulated selectively and intracellular recordings can be made from functionally identified motoneurons. The availability of transgenic and knock-out mice makes it possible to test the role of specific molecules and cell types in generating these specific patterns of connections. The experiments proposed here examine how each of these pathways develops. For the Iα pathway from muscle spindles, we will focus on the role played by the spindle itself in providing trophic support and instructive guidance for Iα sensory neurons, both during embryonic development and during the neonatal period when synaptic connections are still being formed. We will also test if the ETS gene family of transcription factors is involved in helping to specify the pattern of Ia sensory-motor synapses. For the polysynaptic Iβ pathway, we will determine if the projections develop specifically from the outset, as for Iα connections, or if synaptic patterns are remodeled as in other parts of the nervous system. Although the experiments in this proposal are confined to normal development, it is likely that knowledge of the mechanisms involved in the formation of spinal pathways during development will be useful in developing strategies for the reconstruction of these pathways after spinal injury.

PI: Freeman, Lisa
Title: Interdisciplinary Research Training for Veterinarians
Abstract: Well-trained comparative medical scientists are needed to meet the research needs of the 21st Century. Veterinarians currently are underrepresented in biomedical research but can make a unique contribution because of their expertise in clinical practice and fundamental biology, as well as their knowledge of spontaneous animal models of human disease. The goal of the proposed education program is to attract veterinarians to NIDDK-relevant research. This program is focused primarily on veterinary residents, a highly motivated group for which research training usually is not provided. This program is designed to first create the desire to pursue research and then to nurture these candidates with strong mentoring and programs, as well as by providing readily accessible research opportunities. Finally, common barriers to research will be addressed. A major aspect of the program will be biannual symposia on spontaneous animal models of human disease to disseminate information to the scientific community on the vast array of models available and to create new opportunities for collaboration. This program will utilize the strengths and resources of Tufts University to develop an untapped resource for research scientists. This will be achieved under the mentoring of a program faculty that has been recruited specifically to provide an interdisciplinary team of collaborative scientists in a variety of disciplines and that provides experienced and positive role models. The program faculty consists of 17 faculty from 9 departments on three different campuses, providing a network of research training in nutrition and endocrine, digestive, kidney, urologic, and hematologic diseases. Veterinary residents will be actively recruited, with particular attention paid to minority candidates. The program will consist of six parts: 1) Biannual symposia on spontaneous animal models of human disease to provide greater interaction with researchers from other disciplines and to increase opportunities for collaborative research; 2) A multi-function website to facilitate research, including two web-based courses on laboratory techniques and applied statistical methods; 3) A resident research and development (R&D) seminar series that will include topics to foster an interest in research and to facilitate research training; 4) Short-term introductory research electives; 5) Intensive research training electives; and 6) An active mentoring program. The inclusion of a specialist in outcomes assessment on the program faculty will ensure timely and accurate assessment of the program. Short-term evaluations will be used to guide development in the early stages of the program. Mid-range impact of the program will be evaluated by the number of participants, research presentations, grants, and papers. Long-term outcomes will be assessed by the number of Tufts-trained residents that do postdoctoral training and pursue research careers, and by using existing and novel outcomes assessment instruments. These evaluations, continued refinement, and the commitment of the program faculty will help to ensure that this becomes a self-sustaining program.

PI: Freudenreich, Catherine
Title: Stability and Fragility of Trinucleotide Repeats in Yeast
Abstract: Expansion of trinucleotide repeat (TNR) sequences is the causative mutation for a number of hereditary diseases, including myotonic dystrophy, the most common dystrophy in adults, Fragile X syndrome, the most common form of inherited mental retardation, and neurodegenerative diseases such as Huntington's and the spinocerebellar ataxias. The mechanism of TNR instability is interesting both for understanding the etiology and inheritance of the triplet repeat diseases, and for a basic understanding of genome stability in humans. In addition, expanded CGG/CCG and CTGICAG sequences are sites of chromosome fragility, areas prone to breakage in vivo. Chromosome breakage is implicated in the generation of translocations and deletions found in many types of cancer. The aim of this proposal is to elucidate the mechanisms involved in TNR instability and fragility, and determine how these two unusual characteristics are interrelated using Saccharomyces cerevisiae. A novel genetic assay has been developed that produces a selectable phenotype when a TNR tract expands or breaks. This assay will be used to screen for proteins whose over-expression influences TNR expansion or fragility. The proteins found to influence TNRs will be characterized to determine both their normal cellular functions and their influence on repeat maintenance. In addition, the hypothesis that TNR expansions occur by aberrant lagging strand replication will be tested by analyzing tract stability (by PCR) and fragility (by genetic and physical analysis) in specific yeast replication mutants. The role of the G2IM checkpoint in detecting TNR tract damage and preventing chromosome breakage will be investigated by comparing rates of TNR tract breakage in wild-type and cheokpoint-defective cells. Lastly, these analyses will be extended to other types of minisatellite sequences that act as fragile sites in human cells. The proposed experiments are designed to elucidate not only how simple repeats expand to cause human disease, but also the consequences of and cellular response to expanded tracts, with the goal of understanding how genomic instability can affect human health.