Mark Miller - US grants

9353130 Zuazaga The proposal requests funds to establish an Imaging Center for the Institute of Neurobiology. Four instruments to perform major cell image analysis or tissue processing are requested: 1) a photometry/microscope system for quantitative fluorescence analysis, 2) a confocal microscope, 3) a ultramicrotome for serial sectioning, and 4) a three-dimensional reconstruction and morphometry system. The equipment will expand capabilities of eight research projects involving ten of the Institute's resident investigators, four of them Hispanic women, their students, and collaborators from mainland institutions. The development of cell imaging research will not only enhance current research at the Institute but will also further involve minority students in state- of-the-art neurobiological research. ***

Fetal tissues are more sensitive to the effects of chemical and physical carcinogens than are adult tissues. This suggests that the embryos and fetuses of pregnant women are at a greater risk of developing cancers from environmental exposures than is the adult population. It is thus important to gain further understanding of the mechanism(s) that help modulate the fetal organism's response to environmental toxicants, and to determine how the genetic background of the individual fetus can modulate its response to environmental carcinogens. The goal of this research will be to determine the molecular mechanisms governing the pathogenesis of lung tumors in mice differing in their susceptibility to polycyclic hydrocarbon-mediated tumor formation. Bioassays will determine the lung tumor yield following transplacental exposure to 3-methylcholanthrene in mice differing in their inducibility for CYPIA1, which will allow the tumor incidence to be correlated with the metabolic phenotype of the fetus. By analysis of oncogene RNA expression levels and examination of selected oncogenic loci for mutations by the highly sensitive polymerase chain reaction technique, particular patterns of oncogene over/under- expression and/or mutation can be established in the tumor models. It will be particularly interesting to determine whether differences in the metabolic capacity of the fetus toward 3-methylcholanthrene result in differences in the mutational spectrum at the molecular level, possibly as a result of damage occurring to different segments of the same gene or mutations occurring at different genetic loci . This project offers an unique opportunity to demonstrate the important role played by both genetic and environmental factors in determining the susceptibility of the individual to the induction of lung tumors. The data will show how the genetic make-up of the individual can influence susceptibility to cancer as a result of early in utero exposure to environmental chemicals.

9420845 Kraut The electron transfer reaction between cytochrome c peroxidase (CcP) and cytochrome c will be examined. CcP is a 34,000 M.W. monomeric heme enzyme that catalyses the reduction of peroxide to water, using cytochrome c as the electron donor. One step in the catalytic cycle of CcP involves rapid electron transfer between the hemes of CcP and cytochrome c, although the heme edges are separated by at least 17 . The crystal structure of CcP and the complex formed between CcP and cytochrome c have been determined in this laboratory. These crystal structures will be used to guide site-directed mutagenesis experiments aimed at perturbing specific elements of the electron transfer pathway between the two hemes. The effect of specific mutations on structure and electron transfer rates will be evaluated using crystallography and transient spectroscopy techniques. The results will help to define the structural elements that are required for the long distance electron transfer reaction between the hemes of CcP and cytochrome c. %%% All aerobic organisms obtain the energy required for biosynthetic reactions from the conversion of oxygen to water. This is accomplished by a highly ordered enzyme system that transfers electrons from metabolites to oxygen, while preserving some of the energy from this reaction for use in synthetic reactions. The cellular electron transfer system consists of a variety of metal centers that are embedded in proteins. It is clear that the electrons are transferred between the metal centers, and the electron transfer is controlled by the protein surrounding the metal centers, but little is known about how the protein matrix makes these electron transfer reactions efficient and highly specific. As a first step in understanding this process, this investigation will examine how electron transfer is conducted rapidly between the metal centers of twm well-defined protein molecules. The larger goal of the work is to understand how aerobic organisms achieve the efficient and specific electron transfer reactions that are required for survival. ***

During the preparation of meat by conventional cooking, highly carcinogenic heterocyclic amines are generated. These compounds require metabolic activation by the cytochrome P-4501A family of enzymes in order to exert their carcinogenic effects. Transplacental exposure to these heterocyclic amines may play an important role int he initiation of tumors, as studies by this and other laboratories have demonstrated the unique qualities of the fetus that render it particularly susceptible to tumor initiation by environmental carcinogens. The goal of this research will be to determine the transplacental carcinogenicity of a typical dietary heterocyclic amine, 2-amino-3-methylimidazo[4,5-f]quinoline (IQ). This study will assess the target organ specificity and the potential role of Cyp1a1 in the conversion of iQ to proximate carcinogenic metabolites following in utero exposure to the carcinogen. The pathogenesis of the induced tumors will also be examined at the molecular biological level. Bioassays will determine the tumor yield in various organs following transplacental exposure of mouse fetuses to IQ. The ability of IQ to increase the levels of Cyp1a1 will be assessed by biochemical and northern blot assays. Tissue generated from the bioassay will be embedded in paraffin and used to determine the types of genetic damage mediated by IQ at selected oncogenic loci implicated in human cancer, in particular the ki-ras and p53 genes. By analysis of these oncogenic loci for mutations by the highly sensitive polymerase chain reaction technique, particular patterns of oncogene mutation can be established in the tumor models. The induction of tumors following treatment during the transplacental period with IQ will show how chemicals contained in the mother's diet can lead to cancer initiation as a result of the in utero exposure to these dietary carcinogens.

Lay Abstract PI: Miller, Mark Proposal Number: IBN-9722349 Many goal-oriented behaviors are said to be "motivated" as their occurrence depends upon both extrinsic factors (such as the presence of food) and the intrinsic state of the organism (such as the level of hunger or satiety). This investigation relates the activities of specific brain cells and the connections between them to the generation of complex motivated patterns of behavior. The project focuses on the role of chemical substances called neuropeptides, which modulate the activity of these nervous system cells during motivated behavior such as eating. The studies are conducted as part of an undergraduate summer program at the Institute of Neurobiology in San Juan, Puerto Rico so that every step of the research program involves undergraduates. Most organisms, including humans, exhibit a variety of motivated behaviors. This project investigates the fundamental principles underlying the brain mechanisms controlling such behaviors. By coupling the research to the undergraduate summer research program, the project provides students with access to hands-on research opportunities and experiences that were previously inaccessible to students at the host institution.

DESCRIPTION: The applicant will thoroughly characterize the murine mucosal and systemic responses (cellular and humoral) generated by immunization with HIV envelope protein, and the effectiveness of IL-12 as an adjuvant for the production of immunity at mucosal surfaces will also be examined. In Aim 1, the ability to enhance mucosal immunity by co-administering recombinant IL-12 with HIV gp160 via the oral and intranasal route will be assessed. In Aim 2, DNA IL-12 will be delivered either mucosally or remotely by IM or gene gun, and similar characterizations of immune responses will be performed. In Aim 3, experiments will examine the potential dysregulation of mucosal immune responses by immunization with rgp160 in combination with IL-12 protein or the plasmid DNA-IL-12.

DESCRIPTION: The applicant will thoroughly characterize the murine mucosal and systemic responses (cellular and humoral) generated by immunization with HIV envelope protein, and the effectiveness of IL-12 as an adjuvant for the production of immunity at mucosal surfaces will also be examined. In Aim 1, the ability to enhance mucosal immunity by co-administering recombinant IL-12 with HIV gp160 via the oral and intranasal route will be assessed. In Aim 2, DNA IL-12 will be delivered either mucosally or remotely by IM or gene gun, and similar characterizations of immune responses will be performed. In Aim 3, experiments will examine the potential dysregulation of mucosal immune responses by immunization with rgp160 in combination with IL-12 protein or the plasmid DNA-IL-12.

cockroach

neurotransmitters; motor neurons; brain regulatory center; gamma aminobutyrate; motivation; interneurons; aquatic organism; Mollusca; nutrition related tag; immunocytochemistry; field study; electrophysiology;

A grant has been awarded to Dr. Mark W. Miller at the University of Puerto Rico Medical Sciences Campus to establish a Laser Scanning Confocal Microscope (LSCM) facility at the Institute of Neurobiology. This facility will be shared by the investigators and students of the ten laboratories that comprise the Institute. These laboratories utilize a variety of model systems, ranging from synapse development and specification in the cockroach to retinal aging in the human, to address some of the most challenging issues facing modern Neuroscience. While diverse in scope, these research programs are unified by certain methodological considerations and limitations. In particular, they share a need to localize specific proteins, messenger RNAs, or physiological events, within highly complex tissues. The elaborate three-dimensional structures of neurons, their remarkable phenotypic heterogeneity, and their intricate sorting and trafficking capabilities present formidable obstacles to studies requiring the precise localization of fluorescent signals or markers. In many cases, however, the limitations previously imposed by these factors have been overcome by the introduction of the confocal microscope, an instrument that enables investigators to detect and localize fluorescent signals with a resolution that greatly exceeds the capabilities of traditional microscopy.
The research conducted at the Institute of Neurobiology is dedicated increasing our understanding of the structure and function of nervous systems. Specific programs include: (1) Neuropeptide Y and GABA Expression in the Aging Ground Squirrel Circadian System (Dr. N. Lugo), (2) Dendritic Remodeling of Ganglion Cells after Optic Nerve Injury (Dr. R. Blanco), Survival and Regeneration of Human Spinal Cord Neurons (Dr. D. Kuffler), (4) Ultra-fast Imaging of Subcellular Calcium Signals in Vascular Smooth Muscle (Dr. L. Santana), (5) Role of Engrailed Paralogues in Specification of Neuronal Anatomy and Synaptic Connections (Dr. J.M. Blagburn), (6) Cotransmitters and the Regulation of Complex Behavior (Dr. M. W. Miller), (7) Retinal Aging in Human Populations in Puerto Rico (Dr. E. Kicliter), and (8) Functional Role of Metabotropic Glutamate Receptors in a Rhythm-generating Neuronal Network (Dr. W. Krenz).
In addition to the direct enhancement of these research projects, the proposed facility will have a broad impact on the educational goals and capabilities of the Institute of Neurobiology. Numerous graduate students affiliated with the University of Puerto Rico School of Medicine and the Department of Biology at the UPR Rio Piedras campus conduct their research at the Institute. Moreover, many advanced undergraduate students receive research training by participating in an Independent Investigation course, for which they may receive both credit and funding. Finally, the Institute hosts a summer program in Tropical Neuroethology that brings undergraduate students from around the country to Puerto Rico for an intensive 5-week research experience. The creation of a shared Laser Scanning Confocal Microscope facility will clearly enrich these educational programs and enhance the training objectives of the Institute of Neurobiology.

This collaborative project aims to establish a national computational resource to move the research community much closer to the realization of the goal of the Tree of Life initiative, namely, to reconstruct the evolutionary history of all organisms. This goal is the computational Grand Challenge of evolutionary biology. Current methods are limited to problems several orders of magnitude smaller, and they fail to provide sufficient accuracy at the high end of their range.

The planned resource will be designed as an incubator to promote the development of new ideas for this enormously challenging computational task; it will create a forum for experimentalists, computational biologists, and computer scientists to share data, compare methods, and analyze results, thereby speeding up tool development while also sustaining current biological research projects.

The resource will be composed of a large computational platform, a collection of interoperable high-performance software for phylogenetic analysis, and a large database of datasets, both real and simulated, and their analyses; it will be accessible through any Web browser by developers, researchers, and educators. The software, freely available in source form, will be usable on scales varying from laptops to high-performance, Grid-enabled, compute engines such as this project's platform, and will be packaged to be compatible with current popular tools. In order to build this resource, this collaborative project will support research programs in phyloinformatics (databases to store multilevel data with detailed annotations and to support complex, tree-oriented queries), in optimization algorithms, Bayesian inference, and symbolic manipulation for phylogeny reconstruction, and in simulation of branching evolution at the genomic level, all within the context of a virtual collaborative center.

Biology, and phylogeny in particular, have been almost completely redefined by modern information technology, both in terms of data acquisition and in terms of analysis. Phylogeneticists have formulated specific models and questions that can now be addressed using recent advances in database technology and optimization algorithms. The time is thus exactly right for a close collaboration of biologists and computer scientists to address the IT issues in phylogenetics, many of which call for novel approaches, due to a combination of combinatorial difficulty and overall scale. The project research team includes computer scientists working in databases, algorithm design, algorithm engineering, and high-performance computing, evolutionary biologists and systematists, bioinformaticians, and biostatisticians, with a history of successful collaboration and a record of fundamental contributions, to provide the required breadth and depth.

This project will bring together researchers from many areas and foster new types of collaborations and new styles of research in computational biology; moreover, the interaction of algorithms, databases, modeling, and biology will give new impetus and new directions in each area. It will help create the computational infrastructure that the research community will use over the next decades, as more whole genomes are sequenced and enough data are collected to attempt the inference of the Tree of Life. The project will help evolutionary biologists understand the mechanisms of evolution, the relationships among evolution, structure, and function of biomolecules, and a host of other research problems in biology, eventually leading to major progress in ecology, pharmaceutics, forensics, and security.

The project will publicize evolution, genomics, and bioinformatics through informal education programs at museum partners of the collaborating institutions. It also will motivate high-school students and college undergraduates to pursue careers in bioinformatics. The project provides an extraordinary opportunity to train students, both undergraduate and graduate, as well as postdoctoral researchers, in one of the most exciting interdisciplinary areas in science. The collaborating institutions serve a large number of underrepresented groups and are committed to increasing their participation in research.

This is a planning grant for department level reform at the University of Utah, Department of Electrical and Computer Engineering. In this project, the already strong laboratory component within the ECE program at the University of Utah will be enhanced by developing system-level design projects to be integrated within individual courses and also spanning multiple courses. This builds on a particular strength of the department, which already has a top-notch industrially-sponsored senior design sequence a few courses with strong system design projects. Through this change, it is expected to (1) increase student recruitment and retention (including diverse students who might otherwise not have chosen or stayed in engineering), (2) increase student motivation (system design projects are FUN), (3) increase knowledge acquisition and retention (because you can't forget something that you need to integrate at the end of the semester and perhaps next semester too), and (4) develop system-level design understanding (the formal training of which is very limited within ours and other traditional engineering curricula).

The novel aspects of this proposal include the projects themselves, the methods and concepts of integrating multiple classes within the curriculum, and the methods to assess and implement these projects. Undergraduate students, graduate students, and faculty will work together to build this new program within our department.

Our present understanding of several neurological movement disorders relates to pathologies of specific neurotransmitter systems. For example, substantial evidence indicates that the motor deficits associated with Parkinson's Disease result from the degeneration of the dopaminergic nigrostriatal pathway. In Huntington's Disease, motor deficits are associated with degeneration of striatal GABAergic neurons. Our most effective treatments for these disorders rely on agents that are thought to exert their effects on the synaptic signaling mediated by these neurotransmitters. Relating the activities of such neurotransmitter systems and therapeutic agents directly to human behavior, however, poses formidable challenges. The tong-term objective of this research program is to examine interactions between two major neurotransmitters, dopamine (DA) and GABA, in the regulation of feeding behavior. These two "conventional" neurotransmitters are thought to play key roles in the regulation of appetite, satiation, consummatory behaviors, and food reward in species ranging from invertebrates to man. The proposed studies will use an experimentally favorable model, Aplysia, in which it is possible to identify neurons that exhibit a specific transmitter phenotype, and to relate the activity of those neurons to specific behavior patterns. The three Specific Aims focus on the union of these two neurotransmitter systems in five specific identified interneurons in which DA and GABA are colocalized. The proposed experiments will (1) determine the contributions of colocalized DA and GABA to synaptic signaling, (2) explore the roles of DA and GABA in the modulation of intrinsic synaptic plasticities (metaplasticity), (3) investigate interactions between the cells in which DA and GABA are colocalized and other neurons that converge on common postsynaptic targets (heterosynaptic modulation). Recent advances in our understanding of behaviors related to feeding underscore the utility of this approach. In common with most organisms, the ingestive and egestive behaviors of this system are mediated by a single peripheral "physical plant" that is differentially activated by a multi-functional central pattern generator (CPG) circuit. The capability of a single CPG to achieve such motor program switching is often attributable to neuromodulatory cotransmitters that produce broad and coordinated reconfiguration of patterned motor output. Consequently, by increasing our understanding of cotransmission in this system, these experiments can be expected to reveal mechanistic and organizational principles that are applicable to the performance and dysfunction of motor behavior in more complex nervous systems.

The objective of this research is the development and characterization of new silicon devices that should exhibit a variety of magnetic behaviors at will - even though consisting of non-magnetic silicon. Recent theoretical results demonstrated that on a finely-patterned two-dimensional lattice, electrons are confined to intersection sites; at higher concentrations, additional electrons also occupy interstitial sites. Many-body quantum theory predicts that at these differing concentrations, Ferromagnetic, Antiferromagnetic, and several Other Phases will arise, triggered by electron-electron interactions. It should be possible to "switch" between these thermodynamically stable phases merely by changing a control voltage. Such controllable "Spin Lattices" should prove useful in spintronic devices. The approach is to etch lattices into the gate electrodes of silicon metal-oxide-semiconductor transistors. Theoretical and numerical efforts will evaluate their novel electrical and magnetic properties. Importantly, the phenomena should scale up to room temperature as lattice features scale down below 10 nm.

A broader intellectual contribution of this project is the connection of the most basic quantum physics with advanced silicon microelectronics. Successful devices may play an important economic role by replacing transistors when further transistor miniaturization according to "Moore's Law" becomes impractical, potentially finding uses in spintronics, quantum computation, and other applications one can only conjecture at present. This project will train graduate and undergraduate students in scientific and engineering research, with undergraduate students preparing samples and investigating transistors properties for spin lattices. Graduate students will broadly disseminate this knowledge by creating an interactive exhibit in the new "Utah Museum of Science and Technology".

community

Four partner colleges nationwide apply previous results to develop a core manufacturing digital library of online resources, called learning objects. The learning objects reside in the digital library and support advanced electromechanical courses. Progressive indications and observations of enhanced student performance facilitated by the introduction of learning objects in the basic courses holds promise as a strategy to improve the retention of students in the advanced program courses. The digital library for electromechanical technology validates the impact of learning objects on student retention, performance and ultimately graduation.

[unreadable] DESCRIPTION (provided by applicant): Francisella tularensis (FT) is a category A bioterrorism agent and is the etiological agent of a multisyndromic disease known as tularemia. Tularemia is a rare bacterial zoonosis that ranges from a mild lymphadenopathy to a severe form, termed typhoidal tularemia. Typhoidal tularemia results from inhalation of bacteria, carries a 30-60% mortality rate, and is the predominant form of disease that would result from use of FT as a bioweapon. Although antibiotics are available to treat this disease, initial diagnosis is difficult, likely resulting in delay of treatment in the event of a bioterrorist attack. The only current method of prophylaxis is a poorly-defined attenuated live vaccine strain (LVS) which is not currently approved for general use and is unlikely to be licensed in the future due to safety concerns. The goal of the work described here is to develop a safe, effective cytokine-assisted vaccine against FT using a novel vaccine delivery vector based on replication-restricted recombinant vesicular stomatitis virus (?G-VSV). Aim 1 will utilize a combination of proteomic/bioinformatic, expression cloning, and DNA-sequencing methodologies to identify novel T cell-reactive targets of FT. As high priority target proteins are identified, they will be incorporated into Aim 2 studies to determine their value as immunogens in cytokine-assisted vaccines. Aim 2 will test the efficacy of ?G-VSV as a vector for delivery of cytokine adjuvant(s) in combination with exogenous or ?G-VSV-encoded FT immunogens. Both previously-identified and new targets identified in Aim 1 will be evaluated as vaccine immunogens. Immune responses will be quantitatively and qualitatively characterized in a rodent model of tularemia, and challenge trials will be performed to determine vaccine efficacy. As effective vaccine modalities are identified, complementary mixed-modality vaccines will be designed to allow for qualitative manipulation of the resulting immune responses. Bimodal vaccines will be designed to elicit multifunctional (B and T cell-mediated) systemic and mucosal immune responses that are protective against challenge with viable FT. The studies proposed here will provide new information that should lead to the development of a safe and efficacious vaccine for FT. [unreadable] [unreadable]

DESCRIPTION (provided by applicant): Biomedical research has become increasingly dependent upon software and visualization tools to analyze gene and protein sequences. Providing a proper information technology support environment to analyze sequences and store the data produced is essential, but often presents a particular challenge to researchers who have conducting "wet lab" investigations as their main goal. At present, 28,000 researchers and students depend on the Biology Workbench (workbench.sdsc.edu) to meet their needs for sequence analysis software, data storage, and data retrieval. The Biology Workbench provides users free access to web-based environment where users with only an internet browser can conduct analyses, access databases, and store their results. Users have access to 80 sequence analysis functions and 33 remote databases of international significance, and a storage allocation for their personal data. The user base of the Biology Workbench has now grown larger than the original architecture of the Biology Workbench can support, and its rate of growth remains constant. This proposal outlines a methodology to refractor the existing Biology Workbench to create a more modular, more stable platform to meet the needs of Molecular Biologists for analytical tools, databases, and an area to store the results of their work. In its refactored form, the Next Generation Biology Workbench created under this funding will remove the limitation on number of users, create an easily expansible, reusable structure into which community codes can be mounted. It will provide users access to all currently available Workbench tools, and a variety of new enabling technologies, including visualization tools, Web Services, a user configurable interface, peer-to-peer data sharing capabilities, and grid computing capabilities. The proposal outlines a well defined methodology for creating and evolving the software created under this proposal to ensure that it meets the needs of its user base. It also describes a methodology for expanding the user base into new communities.

The AToL (Assembling the Tree of Life ) is a large-scale collaborative research effort sponsored
by the National Science Foundation to reconstruct the evolutionary origins of all living things. Currently 31
projects involving 150+ PIs are underway generating novel data including studies of bacteria, microbial eukaryotes, vertebrates, flowering plants and many more. The data being generated by these projects include and are not limited to: (i) Specimens and their provenance including collection information, voucher deposition, etc.; (ii) Phenotypic descriptions and their provenance; (iii) Genotypic descriptions and their provenance; (iv) Interpretation of the primary measurements including homology ; (v) Estimates of phylogenies and methods employed; and (vi) Post-tree analyses such as character evolution hypotheses.
While the data collection, storage, and dissemination within each projects are well coordinated, there is a critical need to develop the infrastructure to integrate all ATOL data sources, allowing the individual efforts to become multipliers for global hypotheses. Furthermore, as the projects continue to expand and address diverse corners of the Tree of Life, efficient project management will be greatly aided by workflow and data management tools targeted towards the ATOL problem domain. The project will develop new, compact, abstract data models for phylogenetics, leveraging use cases from a broad survey of empirical projects. The integration system will develop novel mappings between different phylogenetic data domains, and allow individual projects to join a network of integrated databases in an incremental manner. The data provenance system, which allows tracking of how each data object was created, will be unique to
systematics data management. The provenance system will not only allow tracking of what kinds of decisions were made in producing a particular tree or a particular column of a data matrix, but will also allow tracking of alternative data lineages such that, for example, different opinions on character homology might be tracked. The results of the research will be delivered in robust software tools that can be used by the entire evolutionary biology community. The study will develop a community-based formal model of data objects used in systematics, primarily through a continuing set of workshops. This activity will not only develop new data management tools, but will also have the effect of synthesizing disparate views of the phylogenetics research domains. The results of the system will be extensible to other domains of evolutionary biology, thereby contributing to the broader mission of evolutionary synthesis. The project will also provide training for the general systematics community in latest database technologies. Finally, by leveraging existing outreach efforts at the Penn Center for Bioinformatics, the project will link to other biological database efforts in genomics and biomedical sciences, disseminating phylogenetic information to the broad biomedical research community.

[unreadable] DESCRIPTION (provided by applicant): The striatum is critically involved in the planning and execution of directed movement, and numerous neurological disorders are closely associated with striatal dysfunction. Despite a detailed understanding of striatal cellular and circuit function, and compelling evidence that striatal function is to a large extent governed by highly structured corticostriatal input, the intracortical mechanisms responsible for shaping and providing this input remain unknown. I propose to elucidate these mechanisms by characterizing the intrinsic membrane properties and morphologies of the two major corticostriatal layer V pyramidal populations, determine whether they, like some other pyramidal cell classes, form subnetworks of preferential homotypic connections, and whether they are subject to differential local inhibition. Results from these experiments will not only clarify the cortical determinants of corticostriatal activity but, by asking whether functionally related layer V pyramidal cells form distinct or overlapping local networks, will also provide insight into the principles of cortical circuit organization. [unreadable] [unreadable] [unreadable]

DESCRIPTION (provided by applicant): The Institute of Neurobiology is a research and teaching component of the University of Puerto Rico Medical Sciences Campus, a minority institution. Its eleven faculty members are engaged in basic research in the neurosciences, and in training students to do so. The present application is to apply for the funds to continue and extend the highly successful program that was begun almost thirteen years ago. Funding under M-RISP has made it possible to provide badly needed administrative support, thus freeing investigators from some of these duties. It is further affording opportunities for selected faculty members to develop themselves as fully competitive members of the world scientific community, as well as role models for their minority students. The objectives of this application are: 1) to provide institutional research development support to enhance the infrastructure that will facilitate development of basic mental health research projects based on molecular, cellular and integrative studies of the nervous system, and to offer advanced training and experience for faculty members and students by supporting workshops and exchange visits between the Institute faculty and experts from other institutions. 2) to provide support for four individual investigator research projects which are basic to the Institute's goal of developing a critical mass of active investigators conducting research in neuroscience. This will enable not only the development of these research areas, but also the training of minority research assistants at the graduate and undergraduate levels; 3) to give an opportunity to minority undergraduate and graduate research assistants to become involved in mental health related research.

Accounting; Anatomic; Anatomical Sciences; Anatomy; CAT Scan, X-Ray; CAT scan; CRISP; CT X Ray; CT scan; Complication; Computed Tomography; Computer Retrieval of Information on Scientific Projects Database; Computerized Axial Tomography (Computerized Tomography); Computerized Tomography, X-Ray; Data; EMI scan; Elements; Femoral Fractures; Femur; Finite Element Analyses; Finite Element Analysis; Fossa; Fracture Fixation; Funding; Goals; Grant; Great Trochanter; Human; Human, General; Institution; Intramedullary Nailing; Investigators; Location; Lower Extremity; Lower Limb; Man (Taxonomy); Man, Modern; Medical Imaging; Membrum inferius; Modeling; NIH; Nail plate; Nailing, Intramedullary; Nails; National Institutes of Health; National Institutes of Health (U.S.); Numbers; Phase; Procedures; Research; Research Personnel; Research Resources; Researchers; Resources; Science of Anatomy; Skeletal Fixation; Solid; Source; Stress; Structure of greater trochanter of femur; Surgeon; Tomodensitometry; Tomography, Xray Computed; United States National Institutes of Health; Variant; Variation; Work; X-Ray Computed Tomography; anatomy; catscan; computed axial tomography; computerized axial tomography; computerized tomography; femur fracture

DESCRIPTION (provided by applicant): Project Summary This application for a Mentored Research Scientist Development Award (K01) describes a training program and research project that provides the candidate with the necessary skills and laboratory-based techniques to conduct clinical research studies and to make the transition to an independent researcher. Candidate: The candidate's long term goal is to develop his own research program focusing on the structural and functional alterations in human skeletal muscle that result from aging, disuse and disease. The candidate is a Research Associate at the University of Vermont with a background in engineering. He has, for the first time, applied the novel technique of sinusoidal analysis to single human skeletal muscle fibers;thereby allowing the first examinations of the molecular determinants of contractile function in humans. Environment: The University of Vermont is ideally suited to the candidate's proposed research and training. It includes a cohesive group of clinicians and basic science researchers engaged in clinical research and muscle physiology that examines function at the whole body, whole muscle, single fiber and single molecule levels. Research: The objective of the proposed research study is to characterize the molecular mechanisms underlying age-related changes in single human skeletal muscle fiber function. We hypothesize that aging impairs single fiber function by: 1) altering myosin kinetics (increasing myosin attachment time and decreasing myosin rate of force production) and 2) decreasing myosin heavy chain content. To test these hypotheses, contractile performance and myofibrillar protein expression from single skeletal muscle fibers will be obtained from young (21-35 yrs old) and elderly (65-75 yrs old) volunteers. The proposed studies will represent the first comprehensive examination of the mechanisms underlying human skeletal muscle contractile dysfunction with aging at the molecular level. Relevance: Understanding age-related skeletal muscle contractile dysfunction at the level of the myosin- actin cross-bridge is a necessary step towards developing more effective pharmacological and lifestyle countermeasures to correct sarcopenia that are directed specifically at molecular defects.

This project is for partial support of the Moluscan Neuroscience 2009 conference to be held February 13-16, 2009 at the Institute of Neurobiology, a free-standing unit of the University of Puerto Rico Medical Sciences Campus. This is the third conference dedicated exclusively to Molluscan Neuroscience since 1997. The other conferences were the Cell and Molecular Biology of Aplysia and Related Invertebrates meetings held at the Cold Spring Harbor Laboratory (CSHL) in 1992 and 1997. This meeting has as one of its goals to enhance interaction among investigators in the field with a format and scale modeled on the CSHL meetings held in the 90's. The meeting will bring together world experts and students in an environment geared towards interaction, participation and discussion of the latest ideas and findings in the field. It will assemble the international community of neuroscientists and students who use molluscan models to investigate a wide spectrum of fundamental neurobiological problems.

Sessions and symposia will address specific thematic areas in which this field has been most influential. These include: Learning and Memory, Central Pattern Generators, Evolution of Neural Circuits and Behaviors, Development, Neuromodulation, Ion Channels, and Signal Transduction. Emphasis will be placed on platforms designed to increase the accessibility of these resources to the broader Neuroscience community. Workshops will be organized to discuss the use of new tools such as the emerging Genome Projects for Aplysia and related gastropods, cephalopod genomics, genomic and microchemical analysis of single cells and cell compartments, and NeuronBank.org initiatives for archiving identified neurons and their homologs across species.

NSF funds will enable multiple educational and outreach initiatives by providing travel, housing, and registration support for graduate students and postdoctoral fellows who could not otherwise attend. Emphasis will be placed on promoting participation of junior investigators, women, local students from Puerto Rico, and members of other groups that are under-represented in this field. This outreach reflects a firm conviction that the greatest beneficiary of such broadened participation will be the field of Molluscan Neuroscience itself. The host institution will provide administrative support and all funds related to the conference venue.

An award has been made to the University of Puerto Rico Medical Sciences Campus to establish an NSF Undergraduate Research and Mentoring (URM) program at the institution, in order to provide opportunities for undergraduate students to obtain research and mentoring experience in the field of neurobiology. Each year, for the first four years of the project, four students will be recruited to participate in the program, and the students will be provided NSF support for a period of two years. Over the five-year period of the grant, a total of sixteen students will participate in the program. Students from groups that are under-represented in neurobiology will be recruited following their second year of undergraduate study. Existing local networking mechanisms will be utilized to identify students from universities throughout Puerto Rico. Students will be engaged in ongoing faculty research programs on a year-round basis, with the objective of getting students to present and/or publish their work. The areas of research cover a broad range of topics, but are unified by the interdisciplinary goal of using the methods of neurobiology to increase our understanding of nervous system structure and function. Each program investigator has considerable mentoring experience, and the structure of faculty research programs enable undergraduate students to become rapidly involved in publication-quality research. The program mentoring strategy consists of personalized "hands-on" technical guidance that exposes students to state-of-the-art physiological, molecular and imaging approaches and equipment. Students are mentored in the scientific method, learning how to formulate hypotheses that are testable and appropriate to the questions they wish to answer. They are involved in all aspects of the research effort, including design of experiments, data collection, analysis, and communication of results. The importance of trust, cooperation, and teamwork within the "laboratory culture" is emphasized throughout the students' training. This individual mentoring experience is integrated within a highly interactive group setting that conveys and instills the excitement and enthusiasm of scientific exploration. Additional information is available at http://www.neuro.upr.edu/nsf-urm.html, or by contacting Dr. Mark Miller, at mark.miller@upr.edu.

DESCRIPTION (provided by applicant): Clinical observations and experimental animal models implicate the spleen as an important site for host immune responses to blood-borne bacteria. How the spleen's unique tissue structure and the diverse mixture of resident myeloid and lymphoid cells work together in vivo to combat infection remains poorly understood. We expect the native tissue environment to regulate immune cell function in complex and profound ways and therefore in vivo imaging approaches are a crucial complement biochemical and in vitro studies. Using two-photon microscopy and a robust histological analysis, we propose to investigate early phagocyte clearance mechanisms, analyze the trafficking of splenic phagocytes and the role of chemokine signaling in spleen remodeling, and identify the antigen presenting cells (APCs) and tissue microenvironment involved in presenting bacteria-specific antigen to T cells. Our overall goal is to understand how spleen structure impacts immune function. Our hypothesis is that early host-pathogen interactions in the marginal zone induce bactericidal responses in some cells, while in others, those interactions trigger cell migration to the appropriate tissue microcompartments for antigen presentation. Our studies will use the well characterized Listeria model of bacterial infection, in which splenic infection leads to T cell priming and induces long-lasting immunity. The proposed studies will enhance our fundamental understanding of how the spleen responds to blood-borne bacteria and provide a framework for future studies of other more virulent human pathogens. PUBLIC HEALTH RELEVANCE: The spleen is the largest collection of lymphocytes in the human body and plays an important role in immunity to bacteria. However, few details are known regarding how resident immune cells work together to initiate the host immune response to infection. We will use cutting edge single-cell imaging techniques to study the capture of bacteria in the spleen and understand how bacterial antigen is presented to T cells.

Abstract: Cultural Shift: Bringing the JOY (Joint Organization to inspire Young people) back to computation
Through this CPATH planning project, Marietta College is exploring effective means of introducing and/or revitalizing computational thinking across disciplines on its campus and among teachers and students in the surrounding K-12 educational community. Led by faculty in computer science, computer information systems, management information systems and mathematics, this project will create a cultural shift among faculty in multiple areas of study. Faculty from these varied departments are serving on the advisory committee for this planning project and have agreed to pilot threads that introduce computational thinking into their courses: physics, biology, education, graphic design, petroleum engineering and theater.

Intellectual Merit of the Proposed Activities
At a small liberal arts institution, the dependence of each discipline on another is perhaps more pronounced than at larger institutions. Rather than creating discipline-specific computer curricula, professors in most other disciplines rely on the computer science department to offer courses that provide a strong base for computational learning. The expectation is that as students develop mastery within their disciplines, they will be able to integrate this knowledge with their computational skills, which will strengthen their ability to manage information, and enable them to transition to increasingly higher levels of discovery and comprehension. The intellectual merit of this CPATH project stems from its focus on revising basic computer courses to emphasize computational thinking, and integrating computational thinking as a method that informs learning across many disciplines.

Broader Impacts of the Proposed Activities
The broader impact of this CPATH planning project is evident in three elements: campus-wide involvement, community outreach, and piloting threads of computational thinking (CT) in non-computer courses. Faculty and staff from any discipline are invited to explore the concept of computational thinking by participating in workshops, attending presentations and/or serving on a CT project advisory committee. Through creative road shows and summer learning opportunities, middle and high school students are being introduced to CT. An introduction to CT is being added to in-service math teachers? workshops at the college. Faculty in at least two and perhaps as many as six other disciplines are piloting efforts to introduce CT within their courses. The anticipated project outcomes include broadening fluency with computational thinking among faculty at Marietta College and among math teachers in the K-12 educational community; revision of two key computer courses to create a stronger alignment with computational thinking for first year computer students; and increasing excitement about and awareness of the myriad uses for computational thinking among K-12 youth. The project?s evaluation process will measure the effectiveness of the various elements of this project and inform the development of a full implementation proposal in the spring of 2011.

DESCRIPTION (provided by applicant): The broad, long-term objective of this research is to understand how neurotransmitter systems control motor activity. The present study will address the mechanistic and functional consequences of signaling by neurons that contain multiple neurotransmitters. It will utilize an experimentally favorable model in which it is possible to identify specific neurons that exhibit a particular transmitter phenotype and to determine the contribution of those neurons to the generation of complex motor patterns. Experiments conducted to date have 1) localized the neurons that contain GABA and dopamine (DA) in Aplysia, 2) demonstrated that the overlap, or colocalization, of these major neurotransmitter systems occurs in only five neurons, all of which participate in the central pattern generator (CPG) circuit that controls feeding, and 3) localized GABA-DA coexistence to identified interneurons that can specify the functional configuration of this multifunctional CPG. Methods integrating neurophysiology, neuroanatomy, and pharmacology will test the central hypothesis of this study: GABA-DA interneurons that are intrinsic to a multifunctional CPG circuit can specify functional motor patterns via modulatory signaling. The proposed experiments address three specific aims that test this hypothesis: 1) determine the contributions of DA and GABA to rapid and slow synaptic signaling by the neurons in which they are colocalized, 2) explore the roles of colocalized DA and GABA in the regulation of multiple forms of synaptic plasticity that these interneurons display, and 3) determine the respective contributions of colocalized DA and GABA to the modulation of intrinsic membrane properties of postsynaptic motor neurons. These studies promise to lead to insights and principles that will have applicability to motor control in more complex brains, including the human central nervous system. In view of the pivotal role of dopaminergic and GABAergic neurotransmitter systems in our present understanding of major neurological movement disorders, these principles should also inform efforts to develop therapeutic and treatment strategies. The developmental objectives of this project will enable the PI to continue his efforts to acquire competitive research support. In view of positive evaluations of recent proposals, it is anticipated that this objective will be achieved during this grant period. Public Health Relevance: Several major neurological movement disorders, such as Parkinson's Disease and Huntington's Disease, are currently attributed to the malfunctioning or imbalance of specific brain pathways. This project will examine the contributions of brain cells that contain specific signaling molecules, or neurotransmitters, to the control of movement. This investigation will increase our understanding of how brain circuits control motor behavior and how major movement disorders result when these circuits are compromised.

This award is funded under the American Recovery and Reinvestment Act of 2009 (Public Law 111-5).

Founded as an Institute within the University of Puerto Rico in 1967 by the renowned neuroscientist José del Castillo, the Institute of Neurobiology (INB) was established with the goal of using simple organisms to understand neural structure and function. UPR is a major minority-serving institution, and the INB graduate student population is predominantly Hispanic. The focus on Poikliothermic model systems holds new significance in furthering the understanding of the impact of climate change (seawater temperature, salinity, etc.) on ecosystems. The institution is focusing on simple organisms, and common interests such as neural plasticity, temperature adaptation, and signaling molecules. Funds are provided to correct significant deficiencies including 1) an antiquated air conditioning system and 2) an obsolete network cyberinfrastructure. The rejuvenation of the INB will significantly improve ongoing and planned research opportunities and create new opportunities for collaborative research. The renovations will have immediate broader impacts including: 1) significant benefit to the minority student body served by the University of Puerto Rico; 2) an innovative approach to "ecological neurobiology" that will provide the first wave of researchers equipped to deal with the emerging and important issues of global climate change; 3) the establishment of a Neurobiology Education Center that will serve to educate teachers, children and the public about the study of neuroscience, and the special relevance of these studies to Puerto Rico's tropical ecosystem.

DESCRIPTION (provided by applicant): Sensorimotor learning allows organisms to interact adaptively with their environment, yet a detailed description of how sensory input modifies motor circuitry during learning has not been developed. Since many movement disorders involve dysfunction of motor planning and disruption of learned motor programs, understanding how motor programs are instantiated in motor circuits during learning is essential. Vocal learning in songbirds is a particularly tractable model of sensorimotor learning since birdsong is an ethological and readily quantifiable behavior, and because the neuroanatomical substrates of song production are identified and accessible. Song learning entails the convergence of juvenile song toward a tutor song, and although changes in song are associated with changes in premotor nuclei, the synaptic and cellular plasticity mechanisms underlying song learning remain unknown. This proposal describes experiments that will combine a form of rapid song learning with chronic in vivo electrophysiology in behaving animals to identify patterns of premotor activity that accompany changes in song, and utilize in vitro whole-cell recording and minimal stimulation to identify the synaptic loci and mechanisms of this plasticity. Results from these experiments will provide an integrated and comprehensive account of vocal motor learning, from synaptic plasticity to changes in circuit dynamics to altered vocal behavior, and thereby give broad insight into how the nervous system learns adaptive and complex behaviors. PUBLIC HEALTH RELEVANCE: The experiments in this proposal will elucidate the detailed changes in brain circuitry that underlie learning of a complex motor task. Many neurological disorders involve dysfunction of brain circuits that control motor behavior, and understanding how motor behavior is encoded by neurons in motor circuits would provide important insight into how these circuits produce disrupted behaviors in disease states.

The goal of this project is to investigate the effects of blast-induced traumatic brain injury (bTBI) using advanced finite element (FE) modeling techniques. This study will focus on the posterior fossa region of the brain (brainstem, cerebellum, great vein of Galen and other vasculature). Previous studies and recent animal model experiments indicate that this important region of the brain sustains damage in bTBI cases. Further, such injuries are consistent with symptoms reported by subjects. An anatomically correct, biomechanically-based, 3-dimensional FE head model will be produced. A detailed FE submodel of the brainstem and veins will be created, which uses the stress/strain distribution from the global head model as the boundary conditions on the separate assembly model. The effects of blast strength and direction will also be studied.

This research is the direct result of the convergence of several important needs in medicine, engineering, education, industry, and the military. If successful, the research benefits will be broad and influence several areas. Early diagnosis and treatment of traumatic brain injury is critical for long term treatment success and even survival. Both can be aided by developing treatment modalities and dose/response relationships that complement the precise injury mechanisms. The work will lay the ground work for the development of better protective equipment for bomb squads, industrial firefighters, and soldiers. For example, head models can be augmented with models of protective equipment or vehicles. Other injury mechanisms (e.g. impacts, flying shrapnel) could also be incorporated into the models. Making these models available in the public domain will be a considerable contribution to the scientific and engineering community. Students and faculty will interact across traditional disciplines, a synergy highly beneficial. The research materials and procedures resulting from this project will be adapted for classroom use.

DESCRIPTION (provided by applicant): The intestinal immune system is charged with the difficult task of protecting a large environmentally exposed surface from potential pathogens, while simultaneously preventing inflammatory responses to innocuous foreign antigen from food and commensal microbiota. Failure to appropriately protect the mucosa can result in life-threatening enteric infection, and failure to control intestinal immune responses results in chronic debilitating disorders such as inflammatory bowel disease. Recent studies determined that the lamina propria (LP) DC population is primarily comprised of dichotomous (CD103+ tolerogenic or CX3CR1 + inflammatory) DCs. This discovery suggests that the balance between tolerance and immunity rests upon which LP DC subtype participates in the immune responses. However a key and missing component is in vivo knowledge of which LP DC subtypes acquire antigen, the anatomical and cellular context of the each LP DC subtypes' interactions with T cells and microbiota, and the outcomes of these interactions on the character of the cellular immune response. The studies outlined in this proposal will harness the complementary expertise of two laboratories to examine the pathways guiding the delivery of pathogenic and non-pathogenic antigens to LP DCs and the outcomes associated with antigen acquisition by LP DC subtypes. The proposed studies make extensive use of cutting-edge two-photon imaging technology to analyze the trafficking and function of DCs and T cells in the intestine of living mice. The overarching hypothesis of this proposal is that antigen acquisition pathways guide immune responses by delivering antigen to specific LP DC subtypes. The studies in Aim 1 will use complimentary in vivo and in vitro approaches to examine the pathways delivering antigen to LP DC subtypes in the uninfected and infected state. Aim 2 will evaluate LP DC subtype specific responses in the presence and absence of infection. Aim 3 will evaluate the capacity of the LP DC subtypes to shape pre- existing T cell mediated immune responses locally within the intestinal lamina propria. Completion of these studies will put forth a new paradigm demonstrating that antigen acquisition pathways are a controlled proximal mechanism guiding immune response toward tolerance or immunity. RELEVANCE: The intestinal immune system must protect us from a wide array of potential pathogens and simultaneously avoid over-exuberant responses resulting in chronic intestinal inflammation. This study will investigate a novel mechanism for maintaining the balance between immunity and tolerance and will offer new avenues to pursue for improved mucosal vaccine therapy and chronic intestinal inflammation.

Human activities are altering the environment at an alarming rate. A multidisciplinary approach is essential to understand the complex interplay between molecular, cellular, and behavioral responses by organisms under these increasingly stressful conditions. The nervous system is the interface between an organism and its environment.

The Puerto Rico Center for Environmental Neuroscience (PRCEN) will combine neuroscience (the study of the nervous system and behavior) and environmental science (the study of local ecosystem environments) to tackle environmental issues in Puerto Rico's tropical setting. The Center will combine neuroscientists from the Institute of Neurobiology and the Dept. of Anatomy of the University of Puerto (UPR) Medical Sciences campus and environmental scientists from the Environmental Sciences Program and the Depts. of Biology and Chemistry of the UPR Rio Piedras. The alliance will bring together cutting-edge techniques normally associated with cellular and molecular neuroscience with expertise in local ecosystems and environmental science to create a novel field that will require participants to move outside of their comfort zones and learn about entirely new areas of research.

Objectives of the center will be to: (1) establish research programs in the new field of environmental neuroscience, (2) enhance research productivity through faculty and infrastructure development, (3) increase the numbers of minority students attaining advanced degrees in interdisciplinary science, and (4) generate community understanding of the work being done in the Center.

The research subprojects focus on four local ecosystems: terrestrial, freshwater rivers, estuaries, and marine systems. The habitats under study are intimately connected: contaminants in the mountains make their way into rivers, pass through the estuaries, and end up in the sea.

The oceans subproject is designed to understand the consequences of environmental pressures on tropical corals, using state of the art molecular-cellular techniques.

The estuaries project will focus on the blue crab, which supports one of the largest fisheries industries in the United States. This project will use high resolution monitoring to track the presence of contaminants and other environmental stressors, and correlate the resulting environmental data with physiological monitoring of heart and endocrine functioning in this crab.

The freshwater studies will monitor contaminants in three representative Puerto Rican rivers. Four animal models (zebrafish, mosquitofish, and two types of prawn) will be exposed to pollutants found in the three rivers, and a range of physiological and behavioral parameters will be examined.

Finally, the terrestrial project will use sophisticated molecular biology and electro-physiology to examine the nervous systems of fruit flies from different habitats in Puerto Rico. The standard laboratory-reared fruit fly (Drosophila) is a prized and widely-used model system in neurobiology laboratories throughout the world. However, there is a paucity of studies examining this animal in the wild, especially with respect to the specific habitats in which they are living.

Intellectual Merit
The conceptual linchpin of the PRCEN is that the nervous system is the interface between an organism and its environment; a multidisciplinary approach is essential to understand the complex interplay of molecular, cellular, organismal, and ecosystem dynamics faced by organisms under the increasingly stressful conditions created by human impacts on the environment. We refer to this approach as environmental neuroscience. The program will be unified by the central hypothesis that a full understanding of the consequences of pollution and climate change requires dialogue between investigators monitoring environmental conditions and organismal biologists using that information to determine how environment affects function.

Broader Impact
The PRCEN center will change the way we look at environmental problems, and will create a new category of scientists prepared for the environmental challenges developing from human activities. The Center will impact a large number of minority students by tapping into the collective student population of over 19,000. Our undergraduate participants will integrate closely with ongoing NSF sponsored mentorship initiatives such as the Lewis Stokes Alliance for Minority Participation, the Research Experience for Undergraduates Program, and the Undergraduate Research Mentoring Program. Our graduate students will have access to broad training here, and will also be given the opportunity to take courses and train stateside at places like the Marine Biological Laboratory in Woods Hole, MA. Finally, our studies will integrate with local organizations such as the San Juan Bay Estuary Program to coordinate community outreach targeting K-12 education.

DESCRIPTION (provided by applicant): The small intestine lamina propria (LP) DC population is largely comprised of two divergent subtypes; CD103+ DCs with homeostatic/tolerogenic properties and CD103- DCs with inflammatory properties. These observations imply a critical role for delivering antigens to the appropriate LP-DC subtype to guide immune responses toward homeostasis or immunity. Recently it was discovered that goblet cells (GCs) act as a passages to deliver soluble luminal antigens to CD103+ LP-DCs. This discovery suggests that GCs play an important and previously unappreciated role in the induction and maintenance of intestinal immune homeostasis. However, mechanistic details are lacking for how this GC function is regulated and how GC mediated antigen delivery contributes to mucosal immune homeostasis. We observed that cholingeric stimuli induce the goblet cell associated antigen passages (GAPs), and that GCs sensitivity to cholinergic stimuli is regulated by sensing the luminal flora. Moreover in the absence of GCs, the cellular intestinal immune compartment is altered, suggesting that GCs play additional roles in shaping the intestinal immune system. Therefore we hypothesize that GCs play a crucial role shaping intestinal immunity through the recruitment and conditioning of LP-DCs, and by regulating antigen delivery in response to intestinal microbiota to promote homeostasis. In aim 1 we will evaluate how sensing the luminal microbiota alters GC sensitivity to cholinergic stimuli using in vivo imaging and ex vivo studies on gnotobiotic mice, specific pathogen free mice, and induced mutant mice strains. In aim 2 we will evaluate the cellular source of acetylycholine inducing GAPs and how it is regulated, using ex vivo approaches and induced mutant mice with cell type specific defects in acetylcholine production. In aim 3 we will evaluate the role of GCs and GAPs in homeostatic immune responses to luminal protein antigens using regimens to induce and challenge oral tolerance. These studies will also evaluate the role of GCs in recruiting and imprinting LP-DCs. Understandning GAP regulation and the role of GAPs in intestinal immune homeostasis may offer new therapeutic interventions for intestinal inflammatory diseases and avenues to optimize oral vaccines.

The University of California, San Diego (La Jolla, CA) has received an ABI development award to create the CIPRES REST API (CRA), a set of enabling public web services for the biology community. The CRA will make it possible for any biologist with modest programming skills to access and run powerful phylogenetic tools on NSF XSEDE supercomputers from within their preferred work environment. The project will embed the CRA into eight distinct software environments that are widely used in the biology community, providing direct access to supercomputing resources from within these familiar working environments. The CRA will make it possible for scientists to analyze much larger data sets than is currently possible without changing their normal work environment or learning to use new software. Community developers will be able to incorporate the CRA tools into their software environments in new and creative ways. The tools created by this project are an extension of the prototype CIPRES Science Gateway, a web application that allows researchers to run large phylogenetic analyses on NSF XSEDE computing resources through a standard browser interface. In its first 30 months of operation, the CIPRES Science Gateway has run analyses for more than 5,000 users, enabled 400+ scholarly publications in biological disciplines ranging from Ecology to Virology, and was used by at least 68 instructors for curriculum delivery.

The impact of CIPRES Science Gateway prototype has been profound because understanding the evolutionary history of living organisms is central to all fields of biological research, and evolutionary relationships are explored primarily by comparing DNA and protein sequences using computationally intensive algorithms. The rate of DNA and protein sequence acquisition is currently increasing more rapidly than Moore's law. While this represents an unprecedented opportunity to clarify evolutionary relationships, it also means routine analyses can no longer be conducted with laptop/desktop computers. Researchers must have easy access to high performance computing (HPC) resources to participate in the discovery process. The CRA will improve the efficiency of access to computational resources by shifting from the rigid browser paradigm of the CIPRES Science Gateway to a pervasive access model, where the phylogenetic tools required for modern biological research are available from within the researcher's preferred environment. The software and techniques created will be completely generic and easily adapted to any area of science where progress is limited by access to computational resources. More information about this project can be obtained from http://www.phylo.org/index.php/portal/ or mmiller@sdsc.edu.

Science Gateways are virtual environments that dramatically accelerate scientific discovery by enabling scientific communities to utilize distributed computational and data resources (that is, cyberinfrastructure). Successful Science Gateways provide access to sophisticated and powerful resources, while shielding their users from the resources' complexities. Given Science Gateways' demonstrated impact on progress in many scientific fields, it is important to remove barriers to the creation of new gateways and make it easier to sustain them. The Science Gateway Platform (SciGaP) project will create a set of hosted infrastructure services that can be easily adopted by gateway providers to build new gateways based on robust and reliable open source tools. The proposed work will transform the way Science Gateways are constructed by significantly lowering the development overhead for communities requiring access to cyberinfrastructure, and support the efficient utilization of shared resources.

SciGaP will transform access to large scale computing and data resources by reducing development time of new gateways and by accelerating scientific research for communities in need of access to large-scale resources. SciGaP's adherence to open community and open governance principles of the Apache Software Foundation will assure open source software access and open operation of its services. This will give all project stakeholders a voice in the software and will clear the proprietary fog that surrounds cyberinfrastructure services. The benefits of SciGaP services are not restricted to scientific fields, but can be used to accelerate progress in any field of endeavor that is limited by access to computational resources. SciGaP services will be usable by a community of any size, whether it is an individual, a lab group, a department, an institution, or an international community. SciGaP will help train a new generation of cyberinfrastructure developers in open source development, providing these early career developers with the ability to make publicly documented contributions to gateway software and to bridge the gap between academic and non-academic development.

An award has been made to the University of Puerto Rico Medical Sciences Campus (UPR MSC) to acquire a state-of-the-art Laser Scanning Confocal Microscope (LSCM). This instrument overcomes obstacles that previously limited the detection and precise localization of fluorescent signals within nervous systems and other complex three-dimensional structures. The LSCM will be housed at the Institute of Neurobiology (IN), a freestanding unit of the UPR MSC. Eleven laboratories that comprise the IN utilize a variety of model systems, ranging from the neuromuscular junction of Drosophila to mammalian neurons in cell culture, to address some of the most challenging issues facing modern neuroscience. The LSCM will benefit the entire neuroscience community of Puerto Rico as well as investigators in other disciplines that require precise spatial localization of fluorescent markers within biological tissues.

In addition to enhancing research objectives, acquisition of cutting-edge imaging instrumentation will significantly impact the educational goals and capabilities of the University of Puerto Rico Medical Sciences Campus. The proposed instrumentation will benefit students associated with two major NSF-supported initiatives: the Undergraduate Research Mentoring (URM) program in Neural Networks and Behavior which supports year-round investigation by undergraduate students in neuroscience labs throughout the island, and the Center for Research Excellence in Science and Technology (CREST) in Environmental Neuroscience which partners neurobiologists with ecologists investigating the effects of anthropogenic factors on nervous system function. Finally, this instrumentation will promote collaborations between the UPR and leading international investigators, providing opportunities for students to receive a part of their training at major research institutions. Such experiences greatly expand their horizons and ultimately lead to broadened participation in the nation's STEM workforce.

A major goal in the field of neuroscience is to understand how the brain evaluates its surroundings and implements a plan of action. Increasing our knowledge about decision-making could ultimately lead to improved strategies for solving problems in a more effective and adaptive fashion. This knowledge should also provide deep insight into certain behavioral and developmental disorders in which decision-making is compromised.

This PIRE project brings together of a consortium of U.S. and international faculty and students on four interdisciplinary subprojects that are unified by the goal of increasing our understanding of brain mechanisms mediating reward and decision processes. Each subproject will partner investigators and students from the University of Puerto Rico (UPR) and/or Oklahoma State University with a team of international researchers from Canada, Chile, Egypt, Italy, and/or Turkey. The Neural Mechanisms of Reward and Decision project will catalyze advances in research and education that could not occur without international collaboration. PIRE workshops and exchanges will cultivate interdisciplinary cooperation and identify common objectives among groups that investigate the role of dopamine in reward and decisions across a broad spectrum of phylogenetic and mechanistic levels. One subproject will study the impact of parasitic infection on neuromodulatory systems that regulate host behavior in a snail-schistosome system using transcriptomics and electrophysiological techniques. A second subproject will examine how isolation stress during adolescence differentially affects male and female dopamine circuitry and resultant learning, memory and behavior in rats. A third subproject will study the role of dopamine on plasticity of foraging behavior in different honeybee subspecies. The fourth subproject will examine the biophysical consequences of repeated exposure to stimulants on the activity of dopamine neurons. The PIRE project will make continuous efforts to integrate across the four subprojects to achieve a broad understanding of neural mechanisms of reward and decision processes.

Student participants in the Neural Mechanisms of Reward and Decision project will conduct research in the labs of international partners as well as in U.S. labs; they will receive mentoring to develop their critical thinking proficiency and enhance their communication skills and professionalism. All mentors possess considerable experience with international collaboration and cooperation which will be shared and disseminated for the benefit of the entire program. This program also responds to the national need to increase diversity in the scientific workforce. The University of Puerto Rico has historically served as a rich source of talented students who pursue graduate degrees in institutions on the island and elsewhere. This program will enhance our ability to provide students with international research experiences that promote their global engagement. A great beneficiary of broadened global participation in the field of neuroscience will be the field itself.

ABSTRACT Core B ? Community Outreach and Translation Core (COTC) The goals of the Community Outreach and Translation Core (COTC) are to transmit research findings that identify risk factors for childhood leukemia (CL) in order to improve environmental health literacy among health care professionals, concerned families, and patient advocacy groups as well as to provide a scientific basis for developing prevention programs for CL. We will communicate findings to health professionals as well as policy and public health authorities. An emphasis will be on developing strategies to reduce environmental exposures that increase the risk of CL and promote children?s environmental health. A special emphasis will be placed on reaching the Latino community. An important focus within these goals is creating awareness of the role of pre- conception and pre-natal exposures in initiating CL and other diseases. We will partner with Commonweal, a non-profit organization, and with the Pediatric Environmental Health Specialty Unit (PEHSU) at the University of California San Francisco (UCSF). We will collaborate with Commonweal to seek additional funding to hold one or more retreats that explore the rationale and options for CL prevention activities. These retreats would include key leaders from the medical, research, public health, childhood cancer advocacy, policy, and other related groups to begin to ?change the conversation?. We will achieve the following specific aims: Aim 1: Develop curricula for health care professionals communicating key findings about childhood leukemia and the environment. This will be presented at regional and national medical conferences, via webinars, and through the ?Story of Health? E-book. A special focus will be adapting these materials specifically for nursing, public health nursing, and nurse practitioner/midwife audiences. Aim 2: Establish a Community Advisory Board that will be representative of the various communities with whom we are partnering. Aim 3: Develop novel training materials in collaboration with Commonweal, CIRCLE investigators, and the Administrative Core that will be used to conduct train-the-trainer sessions with the aim of introducing children?s environmental health into existing pre-marital and pre-family programs for such audiences as religious organizations, community and public health nursing, colleges, and others. Aim 4: Develop specific outreach approaches and materials for the Latino community in collaboration with Commonweal and identify and obtain both traditional and non-traditional media coverage of current childhood leukemia findings. Aim 5: Collaborate with California EPA, Office of Environmental Health Hazard Assessment and the UCSF PEHSU to conduct symposia that will update state, federal, and other scientists and the public on current research findings of the Children?s Environmental Health Research Centers. Participate in the CDC Workgroup on Cancer Prevention Across the Lifespan.

This National Robotics Initiative project will test a series of liquid handling robots in school biology and chemistry classes to determine the range of learning opportunities that can be supported through the instructional use of collaborative robots. Low-cost robots using the Lego Mindstorms platform will be used to implement classroom activities ranging from artistic expression using colorful arrangements of liquids to performing experiments using dilution series, density gradients, and spectral measurements. The aim is to make biotechnology more tangible and relevant to students while supporting interdisciplinary learning as recommended by the Next Generation Science Standards (NGSS). This innovative approach to engaging students in biology and chemistry will be tested with teachers and students in grades 6-12, with a range of user studies being employed to examine learning outcomes and guide the development process. The goal is to integrate liquid handling into educational robotics to enhance current science curricula by enabling deeper inquiry, more variety in learning experiences, and increased attention to interdisciplinary and project-based education.

The research of this project will focus on two themes: students' sense making as they engage in inquiry activities using the platform, and the pedagogical and infrastructural support needed for sustainable deployment and implementation. Multimodal data collected from students running experiments will be combined with traditional qualitative and quantitative methods to answer three primary questions related to the two research themes: 1) How effectively does the system capture the practices and inquiry activities of real scientists using similar tools? 2) How do the affordances identified by the first question map onto the learning goals of engaging in extended inquiry cycles within the context of limited amounts of time available to 6-12 grade science teachers? And 3) What implementation challenges are associated with our proposed curricular distribution model which relies on software and instructions being downloaded and fabricated in local Makerspaces or Lego/Arduino kits being used in schools? The research plan for the project will progress over three years from Microgenetic design and testing during the first year to controlled study of in-class effectiveness during year 2. In the final year of the project, teacher-led in-class effectiveness will be examined.

DESCRIPTION (provided by applicant): The University of Puerto Rico (UPR) Medical Sciences Campus proposes to establish a COBRE Center that will significantly strengthen the research infrastructure of the institution and that will impact biomedical investigation throughout the island. The specific aims of this COBRE Center for Neuroplasticity at the University of Puerto Rico are to: 1) Foster development of junior investigators into competitive researchers working on projects with direct biomedical significance. An intensive mentoring program will be implemented with the goal of creating a culture of research comparable to that of major research-intensive universities. 2) Provide a Neuroimaging and Electrophysiology Facility (NIEF) that will offer state-of-the-art instrumentation, training, and expertise to the neuroscienc community of Puerto Rico. The NIEF will be located in the Institute of Neurobiology, but will incorporate ultra-high-end instrumentation in the new Biomolecular Sciences Building (BSB) after the COBRE has become well-established at the Institute; 3) Partner with IDeA Networks of Biomedical Research Excellence (INBRE) entities in Puerto Rico; 4) Support programmatic activities that increase interdisciplinary collaborations at the basic research level. The major strengths of the proposed COBRE include: 1) the PI, who is both an accomplished researcher with extensive funding and advisory activities within the NIH, and an experienced administrator with a history of successful initiation of research and training programs; 2) the team of young, aggressive and highly committed scientists; 3) enhanced mentorship possibilities provided by the current level of intellectual and scientific accomplishment present at the Institute of Neurobiology; 3) rational planning for the future by deployment of the new BSB as a centralized hub for integrated and collaborative research in the future of the COBRE Center for Neuroplasticity; 4) the full commitment of the UPR Administration toward assuring the stated goals of the project. This UPR COBRE Center should define pathways and benchmarks for basic and translational research across the UPR system for the next decades.

The Centers of Research Excellence in Science and Technology (CREST) program supports the enhancement of research capabilities of minority-serving institutions through the establishment of centers that effectively integrate education and research. CREST promotes the development of new knowledge, enhancements of the research productivity of individual faculty, and an expanded presence of students historically underrepresented in science, technology, engineering, and mathematics disciplines.

With National Science Foundation support, the University of Puerto Rico Medical Sciences Campus will continue development of its Phase II Center for Environmental Neuroscience. The Phase I Center for Environmental Neuroscience was established to bring together scientists from the traditionally separate fields of neuroscience and environmental science, recognizing the fact that the nervous system serves as the interface between an organism and its environment. Phase II Center investigators and students will explore the impact of anthropogenic environmental degradation on nervous systems at the structural, physiological and behavioral levels.

Responses of nervous systems to environmental degradation will be examined in three subprojects partitioned according to tropical habitats: 1) Marine and Estuaries, 2)Rivers and Freshwater, and 3) Terrestrial. Three objectives that encompass the subprojects will identify nervous system responses that will improve risk prediction or development of resilience strategies to diverse environmental stressors. These specific aims will 1) assess effects of environmental contaminants on nervous system structure and function, 2) determine impacts of climatic variation (acidification, temperature changes) on the molecular and cell biology of neurons, and 3) investigate the impact of anthropogenic stimuli (light, sound) on sensory systems and behavior.

Center educational and career development activities include a newly created course in Environmental Neuroscience, a Seminar Series, a Responsible Conduct in Research module, and the Yale Ciencia Academy for Career Development, a year-long on-line program that provides graduate students underrepresented in science, technology, engineering, and mathematics disciplines with opportunities for mentoring, peer support and networking. Institutional partnerships will also provide Center students and faculty with access to training and research programs at institutions of cutting-edge investigation in neurobiology and environmental science.

PROJECT SUMMARY In general, men and women experience differing degrees of age-related decreases in physical function, with women having a greater prevalence of functional limitations and disability. A key predictor of this decrease in functional capacity is the reduction in leg muscle maximal power (product of force and velocity), which can be improved with exercise training. However, the development of exercise interventions to optimally improve skeletal muscle function in older adults has been difficult, in part because we now know that men and women respond differently to the same exercise training stimulus. In fact, the fundamental mechanisms by which habitual exercise improves physical function in older adults are still not well understood. The proposed studies, which build upon our recent work, are designed to address these knowledge gaps by examining the molecular and cellular mechanisms underlying the response to two distinct exercise training paradigms, and determining how these responses differ between older men and women. We hypothesize that molecular, cellular and whole muscle contractile performance will be most improved in men by traditional low-velocity, high-load resistance training, and in women by high-velocity, low-load power training. Moreover, sex-specific structural responses in myofilament remodeling, protein expression and post- translational modifications will explain these sex-specific performance adaptations to each modality. To test our hypotheses, data will be gathered from 50 healthy, sedentary older men and women (65-75 years) prior to and following a 16-week unilateral exercise training program in which one leg undergoes resistance training and the other power training. The Specific Aims of this project are to identify the sex-specific effects of low-velocity resistance training versus high-velocity power training on: Aim 1) skeletal muscle function at the molecular, cellular and whole muscle levels, and Aim 2) protein expression and modification as well as size at the molecular and cellular levels. Our within subject, unilateral intervention design provides a powerful model to minimize the effects of between-subject variability, and our translational approach will take advantage of our unique expertise with state-of-the-art measures from the molecular to whole body levels. Our results will challenge conventional wisdom by determining the sex-specific responses in intrinsic skeletal muscle adaptations to different exercise training programs. We will advance scientific knowledge by providing critically- needed information regarding the specific molecular and cellular determinants that support exercise-induced improvements in muscle performance. This knowledge will have a significant positive impact on the clinical care of older adults by providing novel insight about optimal exercise interventions to improve skeletal muscle function in each sex, and by identifying potential new therapeutic targets for pharmaceutical interventions. Thus, this project is highly relevant to the mission and vision of NIA to support biological research to mitigate conditions associated with aging that may limit health and independence in older adults.

Summary The intestinal mucosa serves as a barrier to infection from pathogens and normal gut flora. This barrier is defended by the mucosal immune system through a variety of innate and adaptive immune mechanisms. However, mucosal immune responses must be tightly regulated to maintain tolerance to resident normal flora and food antigens. Because peripheral tolerance cannot be explained completely by immunological ignorance or negative selection in the thymus, various mechanisms have been proposed for how lamina propria macrophages (MPs) and dendritic cells (DCs) capture and present luminal antigen in the steady-state to promote intestinal homeostasis. Recently, we described a novel luminal antigen transport mechanism that we've termed goblet cell associated antigen passages (GAPs). The role of GAPs in peripheral tolerance vs. immunity is relatively unexplored, but GAPs deliver small soluble antigens preferentially to CD103+ DCs, which have tolerogenic potential. A key question is whether pathogen invasion mechanisms have evolved to target steady-state antigen acquisition pathways (e.g., GAPs) as a strategy to evade the acute immune response. These studies will use a common and often deadly bacterial pathogen, Listeria monocytogenes (Lm), to investigate how initial host-bacterial interactions (minutes to hours) affects mucosal immunity and link invasion pathways to downstream infection outcomes. Our hypothesis is that bacterial invasion via GAPs will lead to inefficient innate and adaptive immune responses and increase bacterial persistence in the host. This hypothesis will be tested in vivo using a murinized Lm strain (Lm InlAMt) mouse that can infect mice orally and complemented by studying Lm InlAWt infection in explanted human intestinal tissues. These studies build on important recent developments including the murinized Lm oral infection model, in vivo two-photon imaging of the intestine and mesenteric lymph nodes, the development of a human explant infection system and state-of- the-art RNA sequencing approaches to characterize epithelial responses. This work is conceptually innovative in that it examines the first few minutes of an infection, which is rarely studied in vivo and impractical to study in human patients, but which may ultimately determine infection outcomes.

Project Summary/Abstract. Understanding the evolutionary history of living organisms is of central importance to every field of biological and biomedical research. Evolutionary information is critical to the discovery process across all biological scales, from proteins to populations. The CIPRES Science Gateway (CIPRES) is a global resource that speeds up inference of evolutionary history from DNA sequence data. CIPRES provides public access to community phylogenetics software run on high performance computing (HPC) resources at no cost to the user. CIPRES allows investigators to access all the capabilities of phylogenetics software efficiently though a browser interface, without having to install the codes, learn the details of schedulers, and construct command lines. CIPRES is the sole public resource for many of these codes, completes analyses 5-30 fold faster than a typical laptop computer, allows users to run many analyses simultaneously, and provides indefinite storage of the results. At present, approximately 2,000 users per month submit 20,000 jobs through CIPRES; they have produced more than 3,000 peer reviewed publications in subject areas relevant to NIH priorities, from enzymology to epidemiology. CIPRES has provided support for research ranging from HIV virus transmission to the discovery of an entire new branch in the tree of life. All of these discoveries happen more quickly because CIPRES provides easy access to HPC resources. The project proposed here will improve the software used to create CIPRES, making it more effective and easier to access. It will provide an environment that allows researchers to easily collaborate, share data selectively, and make their results publicly available. It will expose access to CIPRES services through other important software environments, including Galaxy and Geneious. The project will add a number of user-requested features so work is more efficient, including restarting jobs that have terminated prematurely, transferring large files, and input file validation with automatic configuration of jobs for optimal execution. The project will provide access to many new community codes that have been requested, and those that appear during the project lifetime. Improvements to the interface will make it faster, more intuitive, and useable on smart phones and tablets. The project will also give users ?cloud-bursting? capabilities. Users will be able to submit jobs to a commercial cloud provider on a fee-for-service basis or via NIH commons account when their job is too large for the standard CIPRES community resources. This capability means CIPRES can be scaled and sustained indefinitely for a user population of any size. These improvements (both in capabilities and compute capacity) are expected to greatly expand the number of users who incorporate CIPRES into their day-to-day workflow. The improvements made here will be available to the global research community through release of the underlying software as an open source, distributable package that can be used by any community of practice to access HPC resources. As a result, all improvements created for CIPRES users can be implemented simply and quickly in other online resources for other specific research communities.

Project Summary. The Administrative Core forms the nucleus of the COBRE Phase II Center for Neuroplasticity at the University of Puerto Rico. As such, it will support the three-fold objectives of the COBRE Center: 1) Develop and maintain a nationally competitive biomedical research program focused on the theme of Neuroplasticity; 2) Facilitate the developmental trajectory of junior investigators toward their transition to independent research careers; 3) Support the COBRE NeuroImaging and Electrophysiology Facility (NIEF). The Administrative Core incorporates all programmatic facets of this initiative, including Faculty Career Development Activities (Laboratory Management, Grant Writing, Manuscript Preparation) and Neuroscience Research Infrastructure Enhancement programs (Seminar Series, Neuroscientist-in-Residence, Annual Puerto Rico Neuroscience Conference, Outreach Activities, and Academic Partnerships). The Administrative Core is composed of a team of individuals with proven track records and experience in the operation and management of large multi-faceted research and training programs. Oversight of the Core is the responsibility of the COBRE Associate Director, Dr. Mark Miller. Miller serves as liaison between the program and its Evaluation Module and he coordinates communication with NIGMS officials. The Administrative Support Office will continue to be staffed by a highly experienced and dedicated team including: Ms. Bethzaida Birriel, Grants Administrator; Ms. Brenda Caban, Accountant; and Ms. Sandra Felix, Purchasing Agent. All fiscal matters will be managed at the Institute of Neurobiology, a free- standing unit under the Deanship of Academic Affairs of the University of Puerto Rico Medical Sciences Campus. Institute personnel will handle most aspects of the post-award administration of funds, including hiring of technicians, postdocs, and students. Finally the Administrative Core will coordinate all program appraisal and assessment, including the participation of an independent external evaluation group from Cooperativa de Servicios de Evaluación e Investigación (CoopSEI). The Administrative Core team will organize the Annual COBRE Retreat and Assessment Activity that will enable all stakeholders to meet, exchange ideas, report progress, and plan future strategies. This meeting will serve as the principal forum for developing and disseminating data required for formative and summative evaluation of all COBRE Center research objectives and programmatic initiatives.