John Porter - US grants

Using microcannulation procedures, we propose (1) to investigate under in vivo conditions the mechanisms whereby anterior and posterior pituitary hormones are secreted into pituitary stalk blood resulting in super-high concentrations in stalk plasma of LH, FSH, TSH, prolactin, ACTH, alpha-MSH, and vasopressin; (2) to determine the role of posterior pituitary hormones in the control of anterior pituitary hormone secretion; (3) to investigate the role of thyroid hormones, gonadal steroids, and adrenal steroids in the secretion of pituitary hormones into stalk blood; (4) to investigate the significance of retrograde blood flow in the pituitary stalk in the delivery of pituitary hormones to, and their transport across, the median eminence and hence to the brain; (5) to determine the mechanisms involved in the hypothalamic secretion of LHRH, TRH, and dopamine into hypothalamic-hypophysial portal blood. Using in vitro conditions, we propose (1) to characterize the subcellular compartmentalization of LHRH, TRH, and alpha-MSH in synaptosome-like particles, hereafter called synaptosomes, prepared from hypothalamic homogenates; (2) to investigate the mechanisms involved in the storage and release of LHRH, TRH, and alpha-MSH from synaptosomes; (3) to test the hypothesis that Ca 2 ion in the presence of depolarizing concentrations of K ion causes TRH, LHRH, and alpha-MSH release from synaptosomes by activating guanylate cyclase and/or adenylate cyclase, thereby forming cGMP and/or cAMP; (4) to investigate the role of synaptosomal microtubules, tubulin, and microfilaments in the exocytosis of TRH, LHRH, and alpha-MSH; (5) to investigate the role of prostaglandins, viz., PGE2 and PGF2 alpha, prostaglandin precursors, and phospholipids enriched with arachidonic acid and lysophospholipids in the release of TRH, LHRH, and alpha-MSH from synaptosomes.

The objectives include ascertainment of the qualitative and quantitative alterations in sex hormone production profiles in postmenopausal women and aging men as well as the consequences of these alterations. In postmenopausal women, the dominant aspect of the sex hormone production profile is decreased secretion of estradiol and increased extraglandular formation of estrone. In menopausal women, emphasis will be given to ascertaining the hormonal basis of those events that lead to hot flashes. In aging men, the dominant sex hormone production profile is decreased secretion of testosterone and increased extraglandular formation of estrogen. In both sexes, extraglandular estrogen formation increases with obesity, age, and hepatic disease. The biochemical events that underlie the development of obesity with aging and the role of the adipocyte in the conversion of C19-steroids to estrogens will be examined. The significance of estrogen in the development of androgen-induced prostatic hypertrophy in aging men and the biochemical basis of the potentiating effect of estrogen on androgen-induced prostatic hypertrophy in aging dogs will be studied. The capacity of the hypothalamus to secrete dopamine as a function of aging will be investigated in rats, including the effects of prolactin and serotonin in the stimulation and inhibition, respectively, of dopamine secretion. The role of the cellular internalization of dopamine in the processing of prolactin in lactotrophs will be studied. The subcellular localization in the aging human brain of such bioactive peptides and amines will be examined employing human brain tissue. The subcellular processing of ACTH, including its precursor molecules and metabolites, will be studied.

The potential role of extraocular muscle proprioception in the neural control of eye movements has been obscure. Although the pattern of sensory and motor neuronal innervation of these muscles has been elucidated, relatively little is known regarding the central and peripheral terminations of the extraocular muscle sensory neurones and the types of muscle fibers with which they are associated. The specific aims of the proposed studies are directed toward elucidating the central and peripheral anatomical structures that underlie extraocular muscle sensory innervation in the monkey using light and electron microscopy. The pattern of central synaptic connections of first order muscle sensory neurons will be determined by transganglionic transport of wheat germ agglutinin-conjugated horseradish peroxidase (WGA/HRP). Secondly, these studies will label the sensory nerve terminals with WSA/HRP, transported anterograde from the site of the first order neurons (i.e., semilunar ganglion), in order to inequivocally localize and characterize extraocular muscle sensory receptors. The proposed studies also provide a unique opportunity to examine sensory-motor interactions in the primate oculomotor system. Reciprocal communications between mononeurons and the muscle fibers they innervate serves to maintain the integrity of each of these components of the motor unit. Similarly, trophic interactions may exist between sensory nerve terminals and the particular muscle fiber types with which they are associated. The proposed studies will examine sequential changes in individual exraocular muscle fiber types subsequent to either sensory or motor denervation of the extraocular muscles. Taken together, the proposed studies will begin to define the sensory and motor neuronal interactions that occur peripherally in the oculomotor system. Our long-term goal is to utilize the information obtained from these morhological analyses, combined with the physiological evaluation of functional status using an alert monkey model, in order to comprehend the peripheral changes subsequent to experimentally-induced oculomotor dysfunction.

We propose to investigate the molecular basis of the impaired ability of hypothalamic dopaminergic neurons of the aged brain to synthesize L-dihydroxyphenylalanine (DOPA), to release dopamine, and to respond to prolactin stimulation. We shall investigate the hypothesis that this impaired ability is a consequence of a deficiency of exposed prolactin receptors (PR) exists in the plasma membranes of aged dopaminergic neurons compared to young dopaminergic neurons and that such a deficiency is a result of altered fludity of the lipid membranes of the neurons. To do so, the number of exposed PR in the aging hypothalamus will be evaluated using radiolabelled monoclonal antibodies to prolactin, and the effects of alterations of lipid membrane fluidity on the available number of PR will be investigated. Lipid membrane fluidity will be changed by modifying the cholesterol-to-phospholipid ratio of the membranes. The consequences of such modifications on the responses of aging neurons to prolactin stimulation will be investigated by examining the intracellular events involved in the synthesis of dopamine, the in vitro synthesis (in organ culture) of tyrosine hydroxylase (TH), the transformation of TH from an inactive form through phosphorylation to an active form, the specific activity of TH per mole of the enzyme, the synthesis of DOPA, and the release of dopamine into hypophysial portal blood. Monoclonal antibodies against TH and against PR as well as affinity-purified, monospecific polyclonal antibodies against prolactin will be utilized in these stuides. These proteins will be used in blocking experiments as well as experiments in which isolation and/or quantification of TH and PR are undertaken. The activity of TH in vivo will be assessed by quantifying the rate of synthesis of DOPA in the median eminence. The rate of release of dopamine by hypothalamic dopaminergic neurons will be quantified by determining the rate of release of dopamine into hypophysial portal blood. Using these various approaches, we propose to ascertain the extent to which down-regulation of PR in the hypothalamus of the aging brain is involved in the reduced capacity of the aged brain to synthesize DOPA and release dopamine.

The longterm objective of this research is to define the hormonal, neurohormonal, and neuromodular control of the molecular and cellular mechanisms involved in the biosynthesis and release of hypothalamic dopamine. To achieve this objective, the role of ovarian steroid hormones, prolactin, vasoactive intestinal hormone, opiate-like peptides, serotonin, nerve growth factor, and inorganic cations on dopamine secretion in hypothalamic dopaminergic cells will be studied. The effect of these agents on the level of tyrosine hydroxylase (TH) mRNA, the mass of TH, the in situ enzymic activity of TH, and phosphorylation of TH in the tuberoinfundibular dopaminergic neurons as well as the release of dopamine into hypophysial portal blood will be investigated. TH mRNA will be quantified using a single stranded (32P)-labelled RNA probe complementary to TH mRNA. The radiolabelled RNA probe, cRNA, will be synthesized using TH cDNA, pTH.4, that was subcloned into the Pst I/Eco RI sites of pSP65 vector. The circular plasmid is linearized with Hind III, and the linear DNA templates transcribed with SP6 RNA polymerase. TH mRNA will be quantified using a solution hybridization/S1 nuclease assay and the (32P)cRNA probe. The mass of TH in hypothalamic dopaminergic neurons will be quantified using an immunoblot procedure and densitometric spectrometry. Purified TH will be used as the reference standard. The in situ enzymic activity of TH in hypothalamic dopaminergic neurons will be assayed using an L-dihydroxyphenylalanine (DOPA) accumulation procedure. The activity will be expressed as moles of DOPA synthesized per hr per mole of TH. The release of dopamine will be evaluated on the basis of the rate of release of dopamine into hypophysial portal blood. The findings of this study will provide insight into the mechanisms controlling neurosecretion by dopaminergic neurons of the brain. An understanding of the molecular and cellular events underlying the secretion of dopamine is of foremost importance in the development of an understanding of the development as well as the degenerative effects of aging on dopamine secreting neurons. An inability to secrete sufficient dopamine can result in Parkinsonism, a disease that afflicts one percent of all persons older than 65 years of age.

The role of extraocular muscle proprioception in visual processing and neural control of eye and eye/head movements has remained obscure. In recent years, some fundamental structural characteristics of the extraocular muscle sensory apparatus have been elucidated to the extent that the peripheral receptor apparatus, location of first-order sensory neuron somata, and course and central termination sites of primary sensory neurons are well defined. Furthermore, compelling anatomical and physiological evidence for the interaction of eye and neck muscle proprioceptive signals, potentially functioning in the coordination of eye/head movements, has been obtained. As yet, we have only the most primitive notions regarding the functional role mediated by sensory signals arising from the muscles that move the eyes. While the most profound results of sensory denervation, to date, appear to involve the visual sensory system, only recently have we made any progress into understanding how proprioception is utilized by the oculomotor system. The application represents a continuation of our efforts to determine the anatomical substrate for and functional significance of extraocular muscle proprioception. We are now in the position to apply correlated anatomical and physiological techniques to the problem of extraocular muscle proprioception. The specific aims of studies to be undertaken in the next project period are directed toward: (a) description of the synaptic relationship of first-order extraocular muscle sensory neurons to second-order neurons in the cuneate nucleus, and of the pathways for subsequent distribution of proprioceptive information and (b) characterization of the nature and precision of signals arising from the eye muscle proprioceptive apparatus by recording from single units in the cuneate nucleus of the chronic alert monkey trained to perform eye movement tasks. Taken together, these studies represent a correlated morphophysiological approach toward understanding what has been a relatively neglected component of the oculomotor and visual systems.

A new series of polymers has been discovered based on polyvinylpyridine containing pendent allyl radicals and anions fully substituted with pyridinium cation groups. These "densely charged" polymers show interesting properties. They are remarkably stable. The dry polyallyl anions are not conducting, but the polyradicals have electrical conductivity in the semiconductor range. A new polymeric structure is proposed, which will be prepared and may have interesting conduction and photoconduction properties. The polymers may also have specific ion-exchange properties. The research deals with the preparation and characterization of a number of such polymers with varying degrees of crosslinking. The electrical properties of these polymers will be tested, and an important part of these tests will be their behavior as electrode-coating materials. This grant is being funded under the Materials Chemistry Initiative.

neurons; secretion; aging;

Travel support is provided for distinguished American scientists to attend an international symposium on "Circulating Regulatory Factors and Neuroendocrine Function" in Bratislava, Czechoslovakia. The Slovak Academy of Sciences in this Warsaw Pact nation will provide transportation from Vienna to Czechoslovakia and expenses for the American scientists while attending this meeting. The meeting will focus on the mechanism(s) by which circulating factors derived from the immune system, heart, kidney, adrenals, thyroid gland, gonads and pituitary gland influence the brain mechanisms which regulate neuroendocrine and other homeostatic functions. This meeting is of timely importance because it is now clear that the lifespan and the function of brain neurons are modulated by agents in the blood. While the blood-brain barrier is believed to protect the brain from many direct influences, there is clear evidence that many circulatory agents exert control over the development and maturation of neurons. The meeting will address this crucial problem in a format which provides for integration of data from many different disciplines. The proceedings of this important meeting will published.

High-speed disk storage for a Connection Machine will be provided for researchers at Boston University. This equipment is provided under the Instrumentation Grants for Research in Computer and Information Science and Engineering program. The equipment will be used for several research projects, including in particular: Computational Investigations of Field Theories, Dynamic Micromagnetic Simulations of Magnetic Films, Chaotic Dynamics, Massively Parallel Implementations of Neural Nets and Geometric Surfaces for Manufacturing.

This proposal is for a conference in the area of general relativity. The organizers view this as a chance to bring researchers in classical relativity, astrophysics and quantum gravity together to discuss common problems. In recent years there has been a bridging of the gap between these disciplines and cross discussion is clearly warranted. For the first time results from mathematical relativity are being used in other more familiar branches of physics. These include pulsars, nuclear matter, and gravitational radiation. In the micro domain, the interface between general relativity and high energy physics is an active area of research. Cosmology, quantum gravity, "worm hole" physics etc. have benefitted from ideas originating in mathematical relativity. Similarly, relativity has benefitted from activities in QCD. These developments clearly show the unity of many areas of physics and the need for conferences encouraging cross fertilization. Finally, the organizers wish to use the occasion of the conference to honor Professor E. T. Newman of the University of Pittsburgh, the current President of the 60th International Society for General Relativity, on his birthday.

This award is for the acquisition of networking equipment required for more effective utilization of an existing Connection Machine CM-2. There are a number of research projects using the CM-2 including: visualization of parameterized Julia sets, computational geometry on a SIMD architecture, simulations of magnetic films, simulation of quantum chromodynamics, and multiple target tracking algorithms for radar applications. The effective use of a high speed, highly parallel computer requires high speed data links. This award is for the construction of a fiber optic network linking five sites at Boston University to a large Connection Machine (CM-2). The research supported by this computer includes mathematical, computer science, electrical engineering, physics, and signal processing applications. The award will make the Connection Machine more accessible and better utilize its high speed computing capabilities.

The Virginia Coast Reserve (VCR) is an extremely dynamic, frequently disturbed landscape which is comprised of elements (e.g. grasslands, marshes) that differ in degrees normally associated with biome-level differences. Because the frequency of disturbance at the VCR is so great the types of ecosystem changes that would normally occur across large distances (continents, biomes) and over long periods of time (e.g. glacial and interglacial periods) happen on decadal time-scales. Ecosystem state changes are frequent. The central research theme of the VCR LTER project is the understanding of the dynamics of ecosystem state change, both the transitions among ecosystem states and succession within these states. The basic elements of this LTER's theoretical concept of landscape dynamics are that 1) there are "attractors" in the ecosystem state space within which successional dynamics operate and 2) there are events that produce transitions from one of these attractors to another. The VCR LTER will continue to study succession within several of the more prominent states (Myrica thickets and salt marshes) and the response of these states to disturbance. This will be accomplished by research in the NSF-defined LTER network core areas as well as additional VCR-established areas as well as additional VCR-established areas examining states and state changes that have occurred at the VCR in geologic, historic, and present-day context. The VCR LTER will address the following working hypotheses: 1) The controls on succession within states are the availability of water, the salinity of that water, the frequency of tidal inundation, and sediment deposition; and 2) Gradients in salinity and tidal inundation frequency and thus organic matter accumulation, primary productivity, and nutrients, are a function of the slope, sea level, and astronomical and storm tides. 3) Many ecological state changes are triggered by infrequent, short-duration, intense disturbances such as coastal storms. The products of the proposed work will be the development of a new theoretical base for maintenance and structuring of landscape, further elucidation of controls on ecosystem processes, and new ecosystem and landscape modeling approaches.

Boston University intends to buy a Thinking Machines Corporation Connection Machine, Model CM-5 to enhance Boston University's massively parallel scientific computing environment. This award is for the purchase of a 20 Gigabyte CM-5 disk array unit. The equipment will be used to support research in molecular dynamics simulation of freezing, simulation of neural networks, quantum chemistry calculations, high energy physics particle tracking simulations, plasma simulations and chaos in N-body systems. Computationally-based research which requires high precision solutions of large systems of ordinary differential equations and /or large matrix systems, will be pursued on-site at Boston University in its enhanced massively parallel scientific computing environment. Its acquisition of a Thinking Machines' CM-5 Machine, and from this award, a 20 Gigabyte CM-5 disk array unit, will enable Boston University to support projects in molecular dynamics, neural networks, quantum chemistry, particle tracking and plasma simulation.

DESCRIPTION (Adapted from Investigator's Abstract): The long-term goal of the proposed studies is to understand the cell and molecular diversity of extraocular muscle (EOM) in health and disease and establish the mechanisms that contribute toward eye muscle sparing or targeting in neuromuscular disorders. Specific Aims 1 and 2 address the origins of the EOM phenotype through manipulations of potential regulatory mechanisms at key times in development. The hypothesis that expression of the EOM-specific myosin is extrinsically regulated requiring signaling mechanisms unique to oculomotor motoneurons will be tested using conventional microscopy and sophisticated cell and molecular biology techniques. Tissue-specific regulatory mechanisms are highly pertinent since mutations may contribute toward EOM-specific disorders. The strategy of disrupting or uncoupling sensory-motor interactions during development also is used to examine modulation of the EOM phenotype by the developing visual and vestibular sensory systems that drive eye movements. Specific Aims 3 and 4 represent an important new step toward linking features of this unique phenotype to disease mechanisms. Much of our knowledge of the responsiveness of skeletal muscle to neurogenic or myogenic disease is based upon information from a limb muscle prototype. Disease states interact with well-characterized muscle fiber types to produce patterned pathological changes that are reliably used in diagnosis. Fundamental differences in the EOM phenotype provide a basis for differential response in disease. Specific Aim 3 analyzes (a) morphopathologic changes to better understand the mechanisms responsible for and adaptive changes subsequent to human congenital fibrosis of EOM and (b) a transgenic mouse model with maldevelopment of some oculomotor motoneuron pools to better understand the etiology of and muscle consequences in Duane's retraction syndrome. Preliminary studies have shown that EOM exhibits constitutively high antioxidant capacity. Specific Aim 4 tests the hypothesis that high antioxidant enzyme activity is an important factor in the resistance of EOM to dystrophinopathies. Free radicals are a final common pathway for neuron and muscle pathology in a variety of diseases and insults. Consequently, the relative protection of EOM seen in a variety of insults may be, at least in part, the result of efficient free radical scavenging mechanisms. Taken together, knowledge of EOM properties and adaptive/maladaptive responses in disease will provide a mechanistic understanding of ocular motility disorders that occur in isolation from or in conjunction with those of other skeletal muscles.

We propose to investigate the molecular basis of the impaired ability of hypothalamic dopaminergic neurons of the aged brain to synthesize L-dihydroxyphenylalanine (DOPA), to release dopamine, and to respond to prolactin stimulation. We shall investigate the hypothesis that this impaired ability is a consequence of a deficiency of exposed prolactin receptors (PR) exists in the plasma membranes of aged dopaminergic neurons compared to young dopaminergic neurons and that such a deficiency is a result of altered fludity of the lipid membranes of the neurons. To do so, the number of exposed PR in the aging hypothalamus will be evaluated using radiolabelled monoclonal antibodies to prolactin, and the effects of alterations of lipid membrane fluidity on the available number of PR will be investigated. Lipid membrane fluidity will be changed by modifying the cholesterol-to-phospholipid ratio of the membranes. The consequences of such modifications on the responses of aging neurons to prolactin stimulation will be investigated by examining the intracellular events involved in the synthesis of dopamine, the in vitro synthesis (in organ culture) of tyrosine hydroxylase (TH), the transformation of TH from an inactive form through phosphorylation to an active form, the specific activity of TH per mole of the enzyme, the synthesis of DOPA, and the release of dopamine into hypophysial portal blood. Monoclonal antibodies against TH and against PR as well as affinity-purified, monospecific polyclonal antibodies against prolactin will be utilized in these stuides. These proteins will be used in blocking experiments as well as experiments in which isolation and/or quantification of TH and PR are undertaken. The activity of TH in vivo will be assessed by quantifying the rate of synthesis of DOPA in the median eminence. The rate of release of dopamine by hypothalamic dopaminergic neurons will be quantified by determining the rate of release of dopamine into hypophysial portal blood. Using these various approaches, we propose to ascertain the extent to which down-regulation of PR in the hypothalamus of the aging brain is involved in the reduced capacity of the aged brain to synthesize DOPA and release dopamine.

9411974 Hayden The Virginia Coast Reserve (VCR) is a dynamic, frequently disturbed landscape. The types of ecosystem changes that normally occur across large distances (continents, biomes) and over long periods of time (e.g. glacial and interglacial periods) happen on decadal time-scales. As a result, ecosystem state changes are frequent. The central research theme of the VCR LTER project is the understanding of the transitions among ecosystem states and succession within these states. Our approach will be to continue to study succession within several of the more prominent states (Myrica thickets, forests and salt marshes) and the response of these states to disturbance. The parameters measured are those in the NSF-defined LTER network core areas. In addition, we have implemented manipulative experiments by altering inundation patterns in marshes and altering the availability of fresh water to terrestrial vegetation on a barrier island. The products of the proposed work will be the development of a new theoretical base for maintenance and structuring of landscape, further elucidation of controls on ecosystem processes, and new ecosystem and landscape modeling approaches.

9523469 Rebbi As HPCC technologies, especially parallel computing and high performance networking, become available to much wider segments of the community through the emergence of the NII, there is an increased need for people who understand this technology and are capable of applying it in all sectors of education's, research and business. It is more important than ever to find effective means to link efforts on a national scale to regional and local efforts in order to expand access to and involvement in HPCC for broad sectors of society. The MARINER (Metacenter-Affiliated Resource In the New England Region) project by Boston University will serve as a focal point for HPCC/NII related activities in the New England area. It will act as a catalyst for wide diffusion of technologies and as a channel to the Metacenter. Mariner will draw on the University's advanced computing and communications facilities and build on its experience in computer science, computational science scaleable parallel computing and wide area networking to provide education and tra ining to a wide audience and reach out to institutions within the area that have hitherto not been part of the HPCC/NII community. Key elements of MARINER are: training, education and research programming, outreach programs and activities, and access to facilities at boston University.

9419475 Clarke This project is part of the Southern Hemisphere Marine Aerosol Characterization Experiment (ACE-1), which will be conducted in the remote Southern Pacific in 1995 using aircraft, ship-board, and ground-based measurements of a wide variety of chemical species, physical and meteorological parameters with the central goal of investigating the formation, evolution, and optical properties of atmospheric aerosol. In this component, a variety of instruments will be deployed on the C-130 aircraft to measure aerosol properties: thermally conditioned optical particle counters will be used to assess size resolved composition of the larger optically active aerosol, various condensation nuclei counters, ultrafine condensation nuclei counters, and a differential mobility analyzer to determine the condensation nuclei size spectrum, nephelometers for the aerosol scattering coefficient, and an aethaelometer for the aerosol absorption coefficient. The objective of this work is to characterize aerosol radiative properties in the region studied, and also to develop tools that may lead to a predictive understanding and modeling of the remote aerosol and its radiative properties. The following specific objectives will be addressed: Establish a link between aerosol microphysical, chemical and optical parameters that can be used to model aerosol optical properties, radiative forcing and satellite derived radiance. Examine the contribution of fine particles (e.g. sulfate) and coarse particles (e.g. sea salt) and relative humidity to the local and column aerosol scattering extinction. Develop a quantitative understanding of the processes shaping evolution of the aerosol in the marine boundary layer during Lagrangian advection. Identify the presence of particles recently formed in situ and/or surface derived aerosol (sea salt, dust, soot) in the mid-troposphere and examine the relationship of associated air masses to environmental processes and sources. Work with other ACE-1 participants towa rd linking aerosol data, lidar data, gaseous precursors, etc. on global, regional and local scales to identify key parameters and processes for a dynamic model of remote marine aerosol.

This project concerns the acquisition of a safe, reliable, and cost-effective DNA sequencing instrument that can be used in undergraduate education and research. The evolving biotechnology curriculum in the Department of Biological Sciences has three components: recombinant DNA techniques, the generation of genetic sequence information, and the analysis and manipulation of genetic information. The project focuses on meeting the instrumentation needs of the second component by obtaining a fluorescent-based DNA sequencer that uses a new capillary matrix electrophoresis technology. This instrument meets all of the project's requirements in being safe (no radioactive nucleotides and no acrylamide gels), reliable (fluorescent nucleotide chemistry is a standard technology in the genome projects), and cost-effective (DNA sequencing reactions are performed using a polymerase chain reaction sequencing kit at a cost of about $5 per reaction). All biology undergraduate students can learn how biotechnology is performed by gaining hands-on experience in structured required laboratory class exercises and in doing independent, faculty-sponsored research projects. For students to prepare for the careers of the future and for their taking on leadership roles after graduation, it is imperative that undergraduate students majoring in biology have a firm understanding of what biotechnology is and how this discipline is affecting science and society.

9627624 Hayden The proposed workshop will be held at the University of Wisconsin, Madison, from 18-20 July 1996. The workshop will 1) detail the programmatic similarities and differences in Land-Margin Ecosystem Research (LMER) and the Long-Term Ecological Research (LTER) programs, 2) evaluate the scientific merits for a LTER and LMER merger, 3) evaluate the administrative merits for an LTER and LMER merger, 4) evaluate the special needs of each program, 5) assess the means and procedures for merger of the two programs, and 6) prepare a report to NSF with recommendations concerning the future management of LMER with respect to the LTER program. It is anticipated that the results of this workshop will serve as a means to guide to future directions in long-term coastal research, and promote the synergism between LTER and LMER research programs.

This project will support two planning workshops to facilitate preparation of a Master Plan for the University of Virginia Coastal Research Center (VCRC). The goal of soliciting input from a variety of scientists and field station managers is to make the proposed field station a scientifically attractive place for investigators from a broad range of academic backgrounds as a way of enhancing understanding of coastal systems. The goal of these workshops would be to plan the research/educational infrastructure that will serve the needs of a diversity of scientists from a variety of institutions with a common interest in integrated interdisciplinary research. Ultimately, the product to be derived from the workshops is the Master Plan for development of the site, including enhancement of existing facilities and construction of new, needed infrastructure. The first workshop will be conducted at the VCRC to solicit information about the needs of potential users of the field station and advice from others with experience in development of field stations. Specifics that would be addressed at this workshop would include both Phase I and later phases of the station construction. Workshop participants will be asked to assess Phase I facilities with respect to the functional aspects of accommodating the housing and laboratory requirements of a variety of physical and biological scientists. Additionally, the participants would be asked to provide insight into the type of field instrumentation, data collection activities, data management, and logistical support envisioned for later phases of the VCRC development. The objective of the second workshop will be to prioritize the recommendations of the first workshop. In addition to UVa coastal research scientists, invitations to the second workshop would be extended to non-UVa scientists who are actively seeking funding to conduct work at the VCRC.

The Virginia coast is an extremely dynamic landscape. The Virginia Coastal Reserve (VCR) LTER focuses on understanding the relationships between natural and anthropogenic forces on the ecology of a coastal barrier island, lagoon and mainland system. Frequent storms, tides, and winds cause sea level variations that affect over 70% of VCR's land area. Over the last century sea level rose 35 cm, the highest rise along the Atlantic coast. Seventy years ago the dominant species, eelgrass, disappeared from the lagoons; recolonization began anew in the past 5 years. In addition, 60-90% of the barrier island uplands is new land since 1979. This land creation has left a century-long legacy that we can now use for natural experiments.

The central hypothesis of the new phase of the VCR LTER program is that ecosystem and landscape dynamics and land use patterns within the watersheds of the VCR are controlled by the vertical position of the land, the sea, and the freshwater table surfaces. Our research approach is to utilize short-term manipulative experiments, long-term observations, and computer models. We will also capitalize on nature's "experiments" where new landscapes continue to be created and recolonizing species may be moving lagoons back to conditions of earlier times. We will also expand our research to examine 56 watersheds on the peninsula, which currently vary in cover from 80% forest to 80% agriculture. This research goes to the heart of solving fundamental societal problems associated with environmental change at the land-sea interface.

This award provides support for the construction of a laboratory, a housing unit and a conference center at the Anheuser Busch Coastal Research Center (ABCRC) of the University of Virginia in Oyster, VA. This field station, located close to the point at which the Chesapeake Bay joins the Atlantic Ocean at the southern tip of the Delmarva peninsula, provides researchers and students access to an area where they can investigate the ecological, chemical and geomorphological processes that regulate coastal systems and landscapes. Until recently, the station has had relatively little physical infrastructure. An expanding research user base and the need to develop educational facilities have motivated a facilities development plan of which this project is the first stage. Although a relatively new station, the ABCRC has a strong history of supporting research and training. The proposed improvements will foster a fundamental change in the use of the ABCRC site. Existing facilities are able to support only a short-term presence by researchers at the station. With the proposed laboratory, housing and commons facilities, researchers will be able to spend more time on site, leading to more productive use of field research time. Additionally, graduate and undergraduate courses can be offered at the station once space is available for class and laboratory activities. The planned construction consists of a laboratory (5,000 sq. ft.), accommodations (4,750 sq ft.) and an education and conference building (a 3,500 sq. ft.). The total cost of construction of the three buildings is approximately $1.5 million; the institution will provide the funds required in addition to those provided by the NSF.

The Virginia Coast Reserve (VCR) Long Term Ecological Research (LTER) Program examines the response of coastal barrier ecosystems to climate change, sea-level rise and intensified human use. Studies of watersheds, marshes, tidal flats, bays, and barrier islands show that there are surprising and dramatic changes in ecological dynamics, so-called 'tipping points' or thresholds, where one ecosystem rapidly shifts to another. When these changes occur, many key ecosystem services - storm buffering, commercial fisheries support, carbon sequestration and wildlife habitat - are lost and may be difficult to restore. The VCR LTER uses long-term observations, experiments, and modeling to characterize how, where, and when these tipping points occur, the environmental drivers and biological feed-backs that move ecosystems toward or away from critical thresholds, and the resilience of ecosystems to resist change.

Understanding non-linear responses to global change is critical to managing coastal ecosystems where over half the global population lives, and where humans are altering systems at unprecedented rates. VCR research findings will be broadly disseminated through the program's website, publications and presentations, and partnerships with regional stakeholders to inform policy decisions. The site's Schoolyard Program will provide important training and infrastructure to primary and secondary schools in the local counties, which are some of the poorest in Virginia, and where over half the students are women and minorities. Training of future environmental scientists through the graduate programs at the Virginia Coastal Reserve is one of the priorities of the proposed research.

? DESCRIPTION (provided by applicant): Biliary atresia (BA) is a fibrotic disorder that is the leading cause of neonatal cholestasis and the most common indication for liver transplant in the pediatric population. Although epidemiologic data suggest that BA arises from the interplay of genetic risk factors coupled with environmental exposures, the etiology is unknown. Insight into the pathogenesis of BA comes from the study of a naturally-occurring animal model of the disease. Over the last 40 years, there have been four outbreaks of BA in newborn livestock in Australia associated with ingestion of plants from the genus Dysphania by pregnant sheep and cows. Clinical and pathological findings from the affected lambs and calves show striking similarities with human BA, in particular marked fibrosis at the time of diagnosis. In our original proposal, we proposed to isolate the Dysphania biliary toxin. We now report that we have identified a selective extrahepatic biliary toxin, a previously undescribed isoflavonoid termed biliatresone, and have developed a new model of BA in larval zebrafish. Additionally, we have 1) identified structural features of biliatresone responsible for biliary toxicity, 2) demonstrated tht it binds reduced glutathione, cysteine and histidine in vitro and shown that the binding may be important for toxicity in zebrafish and mammalian cholangiocyte models, 3) identified a genetic link between toxin susceptibility in zebrafish and BA in humans, 4) demonstrated that biliatresone destabilizes mammalian cholangiocyte microtubules and alters cholangiocyte polarity, and 5) shown that biliatresone causes changes in Sox17 in mammalian cholangiocytes that are paralleled by changes observed in human BA livers. We hypothesize that biliatresone-induced toxicity is mechanistically relevant to human BA, and that signaling pathways related to susceptibility loci on human chromosome 10, oxidative stress, cell polarity, and Sox17 are critical to extrahepatic ductal atresia. Our overall goal is to employ a combined chemical, genetic, and cell biological approach to further understand the mechanism of extrahepatic duct obstruction and atresia. There are four specific aims: 1) to determine the effects of biliatresone on redox stress in the neonatal liver and bile ducts; 2) to determine the mechanism of biliatresone-mediated bile duct disruption at a cellular level, specifically to determine the relationship between oxidative stress, microtubules, and polarity in BA, and to identify signaling pathways downstream of Sox17 that could cause BA; 3) to identify genetic modifiers of biliatresone toxicity and determine their relevance to human BA; and 4) to synthesize and conduct structure-function studies of biliatresone and related compounds in order to identify critical structural features underlying toxicity. The proposed experiments will yield novel information about the mechanism of biliatresone-mediated BA; more importantly, they will provide insight into general mechanisms of extrahepatic ductal damage and potential therapies that are highly relevant to human BA.

The transportation sector is in the midst of a major transition to electric vehicles as automotive manufacturers have launched large electric vehicle development programs. Automotive technician training programs will need to adjust to these changes by providing technical education that focuses on electric vehicle technologies. Students need to learn how to diagnose a new set of problems and how to use equipment and tools to maintain these vehicles. This project will develop a new certificate program in hybrid and electric vehicle technology leading to an associate degree in automotive technology. Technician training will focus on the theory, operation, diagnosis, service, and repair of systems related to the propulsion of electric vehicles. Professional development workshops will be provided for postsecondary educators so that they can upgrade their automotive technology programs. During the workshop, participants will learn about battery technology, electric motors, and safety procedures through classroom instruction and hands-on learning experiences with electric vehicles and service equipment. High school students will have the opportunity to learn about careers in automotive technology through summer camps in which students will learn the hands-on basics of vehicle maintenance.<br/><br/>The primary goal of this project is to help develop the technician workforce that can support the new infrastructure for electric vehicles. This project aims to increase the capacity of faculty to prepare automotive technicians to repair and service electric vehicles, the number of automotive technician graduates with the knowledge and skills needed to diagnose and repair electric vehicles, and the number of high school students enrolling in automotive technology degree and certificate programs. Faculty in automotive technology programs at two-year institutions will be recruited to participate in a summer workshop on electric vehicle technologies. Workshop participants will have hands-on learning opportunities focused on diagnostics, testing, and repair procedures. The project will develop a new certificate program that will provide students with advanced training in high-voltage safety systems, removing/replacing batteries, servicing electric drive motors, diagnosing electric power convertors, and other skills needed to service battery electric vehicles. Technician training will focus on the theory, operation, diagnosis, service, and repair of vehicle systems. The project will recruit high school students from traditionally underrepresented groups for automotive summer camps designed to increase students’ interest in taking automotive technician courses. This project is funded by the Advanced Technological Education program that focuses on the education of technicians for the advanced-technology fields that drive the Nation's economy.<br/><br/>This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.