John Williams - US grants

Understanding pancreatic secretion is important both as it relates to the function this digestive organ and in understanding basic cellular mechanisms. Secretagogues such as acetylcholine (ACh) and cholecystokinin (CCK) act on the exocrine pancreas to increase cytoplasmic Ca2+ and release digestive enzymes. The model so far developed and being studied is that receptor activation leads to an increase in the intracellular messengers Ca2+ and diacylgycerol, which in turn activate a battery of kinases and phosphatases which mediate the secretory response. Studies of receptors, intracellular messengers, protein phosphorylation, and amylase secretion are carried out using isolated pancreatic acini. Free Ca2+ in acinar cells will be measured by microspectrofluorometery using the probe fura-2. Kinase and phosphatase activity will be studied as chemical reactions in broken cells. Emphasis will be placed, however, on evaluation of secretagogue regulated substrate protein phosphorylation in isolated acini labeled by incubation with 32P04 followed by 1- and 2 dimensional gel electrophoresis as well as cell fractionation. Measurements of cytoplasmic Ca2+ in single cells w-ill allow determination of whether Ca2+ transients are phasic or tonic and whether individual cells differ or mirror the population. The time course of changes in Ca2+ will be studied to understand the mechanism of maintained secretion and determine if a sustained elevation of Ca2+ is involved. The participation of gap junctions in coordination of Ca2+ between cells of an acinus will be evaluated. Intracellular phosphorylation of individual proteins will be studied as to time dependence, relation to changes in cytoplasmic Ca2+, and the specific kinase involved. Other questions to be studied include: Is the rise in Ca2+ necessary for all changes in protein phosphorylation? Are granule and/or plasma membrane proteins reversibly phosphorylated in a regulated manner in situ? Are particulate proteins reversibly phosphorylated and in what organelle do they reside. Finally, substrates undergoing secretagogue regulated phosphorylation will be purified, sequenced, and used to generate antibodies to better understand their functional role. These studies are also relevant to understanding of the abnormal secretion seen in pancreatic diseases such as pancreatitis and cancer.

Most current designs of total joint replacements make use of bone cement to fix the artificial joint component to the bone. Prostheses which are primarily fixed to the porous inner part of the bone, called cancellous bone, sometimes fail after several years of use, causing much suffering to patients and a high financial cost for additional surgery to remove the failed prosthesis. Loosening of the prosthesis has been indicated as a primary cause of mechanical failure. Stress analysis of prosthesis designs is carried out numerically using the finite element method. To make use of this method, accurate three-dimensional stiffness data are needed for all components of the prosthesis-cement-bone structure. It has been suggested that deeper penetration of the cement into the pores of cancellous bone will enhance the success rate of the prosthesis. This idea creates a layer of a cement-bone composite, the properties of which are not known. In the proposed study the elastic constants of the cement-bone composites will be determined by a pulse transmission ultrasonic technique. Specimens of bovine and human cancellous bone will be cut in the form of 5 mm cubes from larger segments of cancellous bone, after filling the pores of the bone with bone cement. The composite material thus obtained will be assumed to be orthotropic and all nine elastic constants will be determined. This information is expected to be of direct use in finite-element analyses of orthopaedic implants.

This is an application to investigate the control of exocrine pancreatic function at the cellular and molecular level. It includes support for five projects and three core facilities. The first project on calcium and stimulus secretion coupling in pancreatic acinar cells (PI:J.A. Williams) is concerned with the steps by which acetylcholine acts on muscarinic receptors to mobilize cellular calcium as well as the mechanism by which the rise in cytoplasmic calcium leads to changes in protein phosphorylation and secretion of digestive enzymes. The second project on the regulation of pancreatic cells by insulin and islet hormone is concerned with the physiological regulation of exocrine function by islet hormones including insulin, glucagon, somatostatin and pancreatic polypeptide. It also studies the cellular mechanism of insulin action on acinar cells. The third project on the characterization of receptors for cholecystokinin on acinar cells is concerned with the isolation and characterization of CCK receptors, the long term regulation of CCK receptors and biological responses, and the use of pancreatic acini for a bioassay of CCK in plasma. The fourth project on hormonal regulation of pancreatic gene expression will analyze the expression of pancreas specific exocrine genes as well as specific membrane antigens. The fifth project on control of pancreatic fluid secretion will use isolated acinar and duct cells as well as reconstituted cellular monolayers to study the mechanism of secretion of pancreatic juice and its regulation by hormones and neurotransmitters. The support requested is for fundamental research, but the resultant knowledge will be relevant to the understanding of pancreatic diseases including cystic fibrosis, pancreatitus and cancer.

Cocaine is known to act on amine containing neurons to inhibit uptake of transmitters. The reuptake of noradrenaline, 5-HT and dopamine is inhibited, but it is not known whether these mechanisms are equally sensitive to cocaine in intact functioning neurons. It is proposed to compare and contrast the action of cocaine at sites in brain which contain each of these neurotransmitters. Intracellular recordings from neurons in four amine containing nuclei; the locus coeruleus, dorsal raphe, substantia nigra and ventral tegmental area will be made in submerged brain slices in vitro at 37 degrees C. The choice of these nuclei is based on the compact structure, biochemical homogeneity and the presence of amine receptors on the cell bodies of the neurons contained in each of the nuclei. Neurons in each of the four nuclei are similar in that they possess receptors for the same transmitter which they produce, 'autoreceptors'. These nuclei differ however in the type of 'autoreceptor', the number of other amine receptor subtypes and the afferent innervation from other amine containing nuclei. The effects of cocaine on the membrane properties, the actions of exogenously applied substances (noradrenaline, 5-hydroxytryptamine and dopamine) and synaptically released transmitters (noradrenaline, 5-hydroxytryptamine and dopamine) will be determined in these major amine containing nuclei. By investigating the action of cocaine of neurons in four amine containing nuclei the proposed studies will significantly increase our understanding of cocaine actions in the central nervous system. From these studies it should be possible to determine the cellular mechanisms and sites which are involved in the psychotomimetic actions of cocaine, and perhaps in the dependence liability which cocaine is now recognized to possess.

Seventy two teachers, K-12, will participate in a workshop for ten days during the 88 academic year reviewing scientific principles and participating in hands-on science activites using toys. The teachers will be divided in to into four groups for instructional purposes: K-3, 4-6, 7-9, and 10-12. Each teacher will present workshops to other teachers in their school district on teaching science through the use of toys. These workshops will be presented on Fridays and Saturdays between September l987 and April l988. Teachers will be chosen from several school districts in Southern Ohio. They will design strategies to incorporate these activites into their curriculum and increase the participation of girls in science. The university staff and school personnel will evaluate the effectiveness of this program in the class rooms in l988. The participants will be provided with materials to use in their classrooms and to conduct inservice activites. School districts will make commitments to continue this program for at least three years after this program is completed and to provide the support for the local inservice workshops. The PI and staff are eminently qualified to carry out this program. They engaged in a tremendous amout of pre-planning for this project. The university's commitments to this program are unusally strong.

Gastrointestinal hormones are major regulators of the digestion and metabolism of food. Cholecystokinin, probably the most important of these hormones, regulates the exocrine and endocrine pancreas, gallbladder contraction, GI motility, and in addition, is a major neuropeptide. This proposal will continue work characterizing the receptors for CCK and other GI hormones. Specific aims or projects include: 1) purification of the CCK receptor and generation and characterization of antibodies directed against the receptor; 2) characterization of the biosynthesis, processing, and turnover of the CCK receptor. The role of receptor glycosylation in maturation and function will be probed. The regulation of receptor biosynthesis and degradation by CCK and other hormones will be evaluated; 3) the role of distinct receptor affinity states in initiating biological responses will be evaluated using CCK analogy active on only one affinity state; 4) the cellular uptake and processing of CCK will be determined by electron microscope autoradiography and HPLC. Cellular uptake and recycling of the CCK receptor will also be evaluated; 5) other GI hormone receptors particularly for the pancreatic polypeptide peptide, PYY, NPY family will be characterized by ligand binding and cross-linking. The localization of receptors for these and other hormones in intact tissues such as the pancreas and stomach will be evaluated by autoradiography; 6) the mRNA coding for GI hormone receptors will be characterized by expression in Xenopus oocytes. Techniques developed for the CCK receptor mRNA will be extended to other Ca2+ mobilizing receptors. Other receptors will be characterized in oocytes by their electrophysiological effects on endogenous or exogenously inserted ion channels. These projects in total will extend our understanding of the molecular structure and function of GI hormone receptors and the action of GI hormones as physiological and pathophysiological regulators.

This Research Experience for Undergraduates (REU) site project is in the field of chemistry. For a three-year period beginning April 1989, eight (8) undergraduate students will spend ten (10) weeks engaged in basic research in chemistry and, in one case, in a joint chemistry-physics project under the direction of nine faculty members from the chemistry department and one from physics. The program has two main objectives: (a) to develop basic research skills, and (b) to strengthen interest in research and graduate studies. Each student will be supervised by a faculty mentor and will work on an appropriate research project in that faculty member's laboratory, under the direct supervision of graduate students and postdoctoral fellows. In order to encourage student-student interactions, students will be housed together in University dormitories and will be offered formal and informal group activities. All students will take an "Introduction to Research" course, and will present the results of their research at a meeting held at the end of each annual 10-week session.

This proposal combines a number of goals and activities which are among those sought in projects funded under the Private Sector Partnership program. It involves industrial scientists, university faculty, experienced high school teachers as teams of presenters in workshops for 72 precollege teachers. This series of 2-three day workshops extend over two academic years. The first year focuses on applications of science and technology in local industries, and will serve primarily classroom teachers who will develop materials to use in their classrooms. In the second year the emphasis will expand to include aspects of biotechnology, molecular biology in related industries, and the participant pool will be expanded to include science coordinators and science specialists in order to have a broader impact throughout the participating school districts. From these workshops will come a continuing relationship between participants and their industrial mentors from the "focal companies." Cost Sharing by the partners will total 170% of the National Science Foundation grant.

The first aim of this proposal is to determine the direct action of cocaine on the membrane properties of neurons (locus coeruleus (LC), substantia nigra (SN) and dorsal raphe (DR)) in dissociated cell culture. With isolated cells the direct action of cocaine on membrane properties and exogenously applied transmitters can be studied quantitatively since diffusion barriers and endogenous transmitters will be limited. Agonists can be applied in known concentration close to the cell. The effect that cocaine has on the kinetics of ligand activated conductances will be studied to determine if cocaine changes the affinity of the receptor for the transmitter. In addition the use of cultured cells permits whole cell recording with patch electrodes such that more control of the intracellular ion content and second messengers is gained. The proposed experiments are a natural progression of previous experiments in brain slices. The knowledge and experience gained by the work from LC and DR neurons in slices over the past 8 years will be directly applicable to work on isolated cells. The second aim is to determine the changes induced in monoamine neurons recorded in brain slices taken from animals previously treated with repeated injections of cocaine. Chronic cocaine is known to cause long term changes in behavior. The brain slice is particularly will suited for the study of these changes at the single cell level since synaptic connections are left intact and quantitative pharmacological methods can be applied to determine any changes in receptor sensitivity. The long term goal is to use the strengths of each; the brain slice and isolated cell preparations to study action of drugs of abuse at the cellular level. The acute and chronic effects of cocaine will first be studied in cells from relatively homogenous nuclei (LC, SN and DR). Knowledge gained from those relatively simple nuclei will be applied to more complex areas such as the ventral tegmental area, nucleus accumbens and prefrontal cortex. Studies from cells taken from animals which have self-administered cocaine may lead to an understanding of the changes that cause and result from cocaine abuse. Whole cell recordings will also eventually be used to study the changes that occur with other drugs of abuse such as opioids. For example, such recordings from LC neurons with patch electrodes are vital to determine the nature of the reduction in the opioid 'spare-receptor' population following chronic opioid treatment.

The specific aim is to culture dissociated neurons from the locus coeruleus (LC), substantia nigra (SN) and dorsal raphe (DR) and study of the action of cocaine on single isolated neurons. With isolated cells the direct action of cocaine on membrane properties and exogenously applied transmitters can be studied quantitatively since diffusion barriers and endogenous transmitters will be limited. Agonists can be applied in known concentration close to the cell. The effect that cocaine has on the kinetics of ligand activated conductances will be studied to determine if cocaine changes the affinity of the receptor for the transmitter. In addition, the use of cultured cells permits whole cell recording with patch electrodes such that more control of the intracellular ion content and second messengers is gained. The long-term goal is to use the strengths of each; the brain slice and isolated cell preparations to study action of drugs of abuse at the cellular level. In the work proposed here, the acute and chronic actions of cocaine will be determined in cells from relatively homogenous nuclei (LC, SN and DR). The knowledge gained in these simple nuclei will be applied to the action(s) of cocaine in more complex areas such as the ventral tegmental area, nucleus accumbens and prefrontal cortex. Studies from cells taken from animals which have self- administered cocaine may lead to an understanding of some of the changes which cause or result from chronic cocaine use. Whole cell recordings will also eventually be used to study the changes which occur with other drugs of abuse such as opioids. Such recordings from LC neurons with patch electrodes are vital to determine the nature of the reduction in the opioid 'spare-receptor' population following chronic opioid treatment. This award will afford the PI the time to learn and become competent with the tissue culture and whole cell recording methods. The proposed experiments are a natural progression of those which are in progress in the slice preparations. The knowledge and experience gained by the work carried out on LC and DR neurons in the slice over the past 8 years will be directly applicable to work on isolated cells. This coupled with the expertise of members of the Vollum Institute in recording from cultured neurons provide an optimistic setting from which the PI can learn and apply this knowledge to the understanding of cellular mechanisms of drug abuse.

This award will provide funds to the Department of Natural Sciences at South Carolina State College to establish a Research Experiences for Undergraduates (REU) SITE. The focus of the program will be to expand the independent research capabilities of primarily minority undergraduate science majors, provide interactive opportunities for REU students with career research scientists, and to stimulate and prepare students for successful graduate studies and research careers. Research projects will be unified around an aquatic ecology theme, but students will have great flexibility in choosing projects. The locations of cooperating institutions; the Marine Resources Research Institute (MRRI) adjacent to coastal estuarine systems not far from the Edisto River mouth and the Savannah River Ecology Laboratory (SREL) further inland near upper coastal plain habitats will provide students with a comprehensive picture of the kinds of research taking place along the length of an entire river system. Additional training benefits to students will be derived from a series of ecological excursions guided by off-site career researchers in cooperation with SREL and MRRI. This will not only help students improve their concepts of ecosystem interactions, but also provide for increased awareness of career options and graduate studies through interactions with host scientists. A guest researcher seminar series will be held at SCSC to introduce students to outside researchers and students will present seminars of their findings.

In a classical series of experiments, Martin and coworkers described the responses of the chronic spinal dog to several classes of opiate compounds (1-2). From these responses he described three types of opiate receptor designated mu, kappa, and sigma. The sigma receptor was proposed to account for the response observed with the experimental drug, SKF10047. This benzomorphan opiate produced an increase in respiratory and heart rate, mydriasis, and a striking behavioral syndrome suggesting a "canine delirium". The sigma-binding site has now been shown to be a unique, non-opioid binding protein present in the central and peripheral nervous system. Compounds that have high affinity for this site such as haloperidol, phenothiazines and even SKF10047, have interactions with other receptors making it difficult to accurately assess the physiologic and pharmacologic role of the sigma-binding site. Recently several high affinity ligands have become available that are selective for the sigma-site. This should make it possible to study the cellular effects of the interaction between sigma-ligands and the binding site. In the present proposal the action of sigma-ligands on single neurons in the locus coeruleus and dorsal raphe will be studied using intracellular recording in brain slices. The action of sigma-ligands on resting properties, spontaneous activity, membrane currents, synaptic potentials and ligand activated currents will be studied. Studies of specific intrinsic and ligand activated membrane currents in isolation will be carried out with the single electrode voltage clamp. As there has been little work done with selective sigma-ligands at the cellular level, this study will lead to an understanding of the cellular actions of this class of psychotomimetic compounds. Equally important, the results may lead to a better understanding of the pathogenesis of and new therapies for schizophrenia.

This project aids in the implementation of an undergraduate interdisciplinary curricular program in design with plastics and composites. The program is intended for majors in mechanical, chemical and materials engineering and in chemistry. A major component of the curriculum program is the development of a new laboratory course in Practical Polymer Processing, which includes mechanical and structural design, material and manufacturing process selection, mold construction, component manufacture and final product functional testing. Included in the course is a series of processing experiments designed to introduce students to essential elements of polymer and composite processing appropriate to each discipline.

A group of 18 institutions is working together to introduce students to realistic problems and to the technology (graphing calculators and microcomputers) that is capable of dealing with them. Existing materials are being adapted and will be implemented on a state-wide basis. The laboratory materials developed at the University Hartford and the text materials being developed by the Core Calculus Consortium (lead by Harvard University) are being integrated into the new course.

Pancreatic secretion of digestive enzymes plays an essential role in the digestion of food. Digestive enzymes synthesized by acinar cells are packaged into secretory granules termed zymogen granules. Upon stimulation of the pancreas by gastrointestinal hormones and neurotransmitters the zymogen granules fuse with the luminal membrane of the acinar cell such that the granule contents are released. Little is known about this process at the molecular level. Because protein-protein interactions are almost certainly involved in the fusion of the zymogen granule membrane with the plasma membrane it is essential to identify and characterize zymogen granule membrane proteins. Zymogen granules can be highly purified on Percoll gradients, their membranes isolated and the component proteins resolved by 2-dimensional gel electrophoresis. Proteins will be characterized by probing blots of gels with stains, antibodies, lectins and 35S-GTP. Preliminary work has shown multiple yet unidentified small G proteins firmly associated with the granule membrane. The topology of the proteins will be studied by biotinylating or otherwise labeling the outside of intact granules prior to lysis and in the intact cell using Ab against component proteins. N-terminal microsequence will be obtained from blots and used to predict nucleotide sequence to clone by PCR and/or library screening. Preliminary work has obtained partial amino acid and nucleotide sequence for a glycoprotein termed GP-3. Antibodies will then be generated to synthetic peptides or cloned proteins expressed in E. Coli. Ab and cDNA probe will be used for immunocytochemistry to confirm granule membrane localization and to establish whether the protein is unique to pancreas or common to other secretory granule membranes. Ab or synthetic peptide sequences to proteins on the external face of the granule will be evaluated in permeabilized pancreatic acini for their ability to inhibit secretion. Pancreatic AR42J cells and AtL-20 cells will be transfected with sense and antisense constructs to regulate expression of granule proteins and evaluate effects on packaging of granule contents and secretion.

The experiments in this proposal are aimed at uncovering adaptive changes in neuronal activity that result from chronic opioid treatment. Behaviorally it can be observed that a high degree of tolerance to and dependence on morphine develops with chronic use. It is curious that, at the cellular and biochemical level, the degree of tolerance and dependence is orders of magnitude smaller. The disparity in the observations may suggest that there are multiple mechanisms arranged in series that could account for the high level of behavioral tolerance. Another possibility is that regulation of ion conductances other than those directly affected by opioids are altered with chronic treatment and they may account for the increased level of tolerance and dependence. Opioids activate potassium channels and/or the inactive calcium channels to perturb cell excitability and function. In the continued presence of opioids, tolerance develops to these direct actions of opioids. There are, in addition, other adaptive changes in cell function and excitability not directly related to the specific ion channels gated by opioids. The regulation of those channels 'indirectly' gated by opioids is the subject of this proposal. Intracellular and whole cell recordings from neurons in the locus coeruleus and substantia gelatinosa of the spinal trigeminal nucleus will be made in brain slice preparations. With these two preparations, two opioid mediated responses will be investigated, the increase in potassium conductance and the presynaptic inhibition of glutamate release. One consistent observation made with chronic opioid treatment is an increase in the basal and stimulated level of cAMP. In this proposal, the effects of cAMP-dependent processes on transmitter release and ion conductances gated by voltage, transmitters and G- proteins will be identified and characterized. These cAMP-dependent processes will be studied after acute application (min or hours) of opioid agonists and in animals treated chronically (days) with morphine. By investigating the alteration in the regulation of ion conductances not directly affected by opioids, it may be possible to clarify the mechanisms by which the high levels of tolerance found at the behavioral level are generated.

Understanding pancreatic secretion is important both as it relates to the function of this digestive organ and in understanding basic cellular mechanisms. Seratogues such as cholecystokinin (CCK) and acetylcholine (ACh) act on the exocrine pancreas to increase cytoplasmic Ca2+ and release digestive enzymes. There is good evidence that the increase in Ca2+ is both necessary and sufficient to drive enzyme secretion. However, there is still imperfect understanding of how free intracellular Ca2+ is controlled and how Ca2+ acts. In this proposal Ca2+ will be measured by dual wavelength fluorescence digital imaging. The mechanism of Ca2+ oscillations in time and Ca2+ waves around an acinus in space will be investigated. The role of Ca2+, IP3 and gap junctions will be evaluated using both focal extracellular stimulation and microinjection. We have also established the existence of [Ca2=+]i oscillations which doe not involve IP3. We will also investigate the importance of two Ca2+ activated enzymes known to be present in pancreas in mediating the action of Ca2+. These enzymes are protein phosphatase 2B (PP2-B) and calcium calmodulin kinase II. In both cases molecular biological techniques (PCR phosphatase 2B (PP2-B) and calcium calmodulin kinase II. In both cases molecular biological techniques (PCR and library screening) will be used to establish the molecular form present in acini. The properties of native and recombinant enzymes will be characterized and antibodies to the proteins generated. Substrate proteins dephosphorylated or phosphorylated by these enzymes will be identified. In both cases we will establish protocols to inhibit the enzyme to establish its importance in regulation protein phosphorylation and secretion. The immune suppressants cyclosporin A and FK506 will be used to inhibit PP2- B. The kinase inhibitor KN-62, autoinhibitory domain peptides and antibody will be evaluated as inhibitors of CaM Kinase II. In addition to biochemical measurements of secretion in intact and permeabilized cells, we will utilize capacitance measurements on single patch clamped acinar cells to study the temporal pattern of secretion and its relation to Ca2+ oscillations. These studies using patch clamped cells will also allow introduction of peptides, proteins, and Ab to ascertain the importance of PP2-B and CaM Kinase II in secretion. These studies are also relevant to understanding of the abnormal secretion seen in pancreatic diseases such as pancreatitis.

9353938 Williams As a result of the calculus reform movement good curriculum materials which involve the use of technology, modeling, and problem solving are now widely available for the first two semesters of calculus. Similar materials are also becoming available for courses which lead into calculus like precalculus and college algebra. In this project a coalition involving both two- and four-year colleges and universities in Connecticut is working to integrate the materials and approaches so that students taking a reformed precalculus course can move into a reformed calculus course with an expectation of continuity. It is particularly important both two-year colleges and four-year colleges and universities are involved in this process because many students take precalculus at two-year colleges and then transfer to a four-year institution for calculus. Three different workshops are being held in each of two summers on precalculus and algebra, first two semesters of calculus, and calculus of several variables and differential equations for instructors at two and four-year colleges and universities. In addition, key faculty at 7 institutions are identifying and evaluating materials for precalculus and calculus and developing further materials which assure a smooth transition of students between the identified courses. ***

The specific aim is to culture dissociated neurons from the locus coeruleus (LC), substantia nigra (SN) and dorsal raphe (DR) and study of the action of cocaine on single isolated neurons. With isolated cells the direct action of cocaine on membrane properties and exogenously applied transmitters can be studied quantitatively since diffusion barriers and endogenous transmitters will be limited. Agonists can be applied in known concentration close to the cell. The effect that cocaine has on the kinetics of ligand activated conductances will be studied to determine if cocaine changes the affinity of the receptor for the transmitter. In addition, the use of cultured cells permits whole cell recording with patch electrodes such that more control of the intracellular ion content and second messengers is gained. The long-term goal is to use the strengths of each; the brain slice and isolated cell preparations to study action of drugs of abuse at the cellular level. In the work proposed here, the acute and chronic actions of cocaine will be determined in cells from relatively homogenous nuclei (LC, SN and DR). The knowledge gained in these simple nuclei will be applied to the action(s) of cocaine in more complex areas such as the ventral tegmental area, nucleus accumbens and prefrontal cortex. Studies from cells taken from animals which have self- administered cocaine may lead to an understanding of some of the changes which cause or result from chronic cocaine use. Whole cell recordings will also eventually be used to study the changes which occur with other drugs of abuse such as opioids. Such recordings from LC neurons with patch electrodes are vital to determine the nature of the reduction in the opioid 'spare-receptor' population following chronic opioid treatment. This award will afford the PI the time to learn and become competent with the tissue culture and whole cell recording methods. The proposed experiments are a natural progression of those which are in progress in the slice preparations. The knowledge and experience gained by the work carried out on LC and DR neurons in the slice over the past 8 years will be directly applicable to work on isolated cells. This coupled with the expertise of members of the Vollum Institute in recording from cultured neurons provide an optimistic setting from which the PI can learn and apply this knowledge to the understanding of cellular mechanisms of drug abuse.

9354378 Sarquis Faculty at two two-year campuses are revising the General Chemistry course to make it more interesting, relevant, and accessible to students with various academic backgrounds. This effort includes the design, development, and testing of discovery-based laboratory scenarios and take-home lecture supplements that illustrate topics in Chemistry through activities beyond the classroom. In addition to demonstrating general concepts, the activities also involve students in critical-thinking and group problem-solving skills used by professional chemists in industry and academia. To further ensure success, the project is built on the pedagogical foundations of constructivism, Science-Technology-Society, and cooperative learning. Following pilot implementation and revision, these advances in the General Chemistry curriculum will provide a model for adaptation at other two-year colleges and small four-year schools. ***y

Distributive theories, developed to explain Congressional decision making, predict that legislatures whose members are elected in single-member district representational systems will attempt to return policy benefits to their districts in order to gain re-election. Alternative representational schemes are predicted to produce alternative types of policy distributions. This project investigates the connection between the type of election system cities use and the distribution of capital improvement projects. Using a four city sample, this research will evaluate the distribution of expenditures in two single-member district and two at-large electoral systems. Topologically Integrated Geographic Encoding and Referencing (TIGER) files together with automated Geographic Information Systems (GIS) software will be used to map the location of capital improvement expenditures and other critical variables to intra-urban regions. This project will aid in understanding the policy implications that result from the choice of electoral systems in U.S. cities. It should also demonstrate the usefulness of GIS for exploring key hypotheses in urban politics.

9419244 Jouse While the detailed mechanisms for storing memories in the brain are still unknown, neurobiologists have identified specific sites where information is stored. One of these memory sites is the mammalian hippocampus. With this award, a collaborative team consisting of nuclear systems engineers and a neuroscientist will conduct theoretical modeling studies to test ideas about how information is stored in the hippocampus. These studies will provide new ways of thinking about biological memory formation, that will impact future work on the cellular mechanisms of learning, and related engineering applications such as neural network models of learning systems.

9510553 Williams This doctoral dissertation research project investigates the causes and implications of the distribution of land use "bads" through zoning in cities. "Bads" are defined as land uses that produce negative spillovers to surrounding property. Using data from cincinnati, Columbus, Indianapolis, and Milwaukee, the project investigates the effects of electoral institutions on the distribution of land use bads and the politics of bureaucratic/legislative relations between the planning department and city council in distributing these policy outputs. The research also explores the efficiency implications of the distribution of land use bads. Efficiency of bad siting patterns is measured in two ways--the effect of bad siting patterns on property values, and the effect of bad siting patterns on the supply of public goods. This study is innovative and contributes to a larger theory of public policy distribution in two respects. First, it examines the distribution of bads instead of the more often-studied goods. Second, by studying distribution in cities, it is possible to take advantage of institutional variation that does not exist in the study of distribution in the United States Congress, where institutional structures are fixed. ***

9512599 Weete The ion accelerator laboratory at the Nuclear Science Center at Auburn University will be upgraded through acquisition of a tandem accelerator equipped with a radio frequency (rf) exchange ion source, a high voltage source injector system, and a high energy quadrupole focusing lens. The new machine replaces a twenty-six year old accelerator which is nearing the end of its serviceable life. The accelerator laboratory functions primarily in support of various materials research programs in different colleges and departments. These efforts focus primarily on thin film systems and interactions, such as silicon carbide and III-V compound semiconductor epitaxial film characterization, ion- induced radiation damage in insulators, materials oxidation processes, and electronics packaging and diamond film technology. Analysis techniques available in the accelerator laboratory for materials studies include proton induced x-ray emission (PIXE), Rutherford backscattering spectrometry (RBS), light ion channeling (LIC), fast neutron activation analysis (FNAA), and some ion implantation capabilities. Most of these techniques will be available in the upgraded facility, and the new machine will facilitate development of expanded analysis capabilities such as neutron activation/time-of-flight techniques. %%% The new tandem accelerator will allow the laboratory to provide efficient research support for materials science programs through improved access to modern analytical techniques. It replaces an ancient unit which currently consumes large material and human resources for its maintenance. The accelerator laboratory has a long history of collaborative research with nearby minority institutions, and the program has a long-term commitment in support of educational opportunities for minority students in the physical sciences which the new accelerator will enhance. ***

The overall objectives of this research are to describe and understand the chromosomal changes which may be occurring in the Raga pipiens karyotype during normal embryogenesis, the development of neoplasia and the temperature the chromosomal incorporation site(s) of the Lucke Tumor Herpesvirus (LTV) genome. The genomes of embryonic cells, some highly differentiated cells, and Lucke' tumor cells can be "reprogrammed" by nuclear transplant to develop along pluripotent pathways. These unusual properties of Raga pipiens and its associated Lucke' tumor make it an excellent system for analysis of the chromosomal modifications, and their stability, occurring during normal and neoplastic development. An understanding of such modifications will enhance our knowledge of how a common, although pluripotent, genome can give rise to the diversity of cellular phenotypes observed in the vertebrate organism. This goal will be achieved by attainment of four specific aims:(1) continuation of the current efforts to describe the banded karyotype of PNKT-4B and Lucke' tumor cells, (2) development in this laboratory of the capability to propagate the Lucke' tumor, (3) examination of the stability of normal and neoplastic karyotype during "reprogrammed" embryonic development following nuclear transplant, and (4) determination of the chromosomal site(s) of LTV genome incorporation by FISH. Chromosomal loci of NORs, N-bands, are visualized by staining with acidic colloidal silver nitrate and C-bands are induced y hot alkali treatment and Giemsa staining. Elongated prometaphase chromosomes from MTX synchronized cultures can be G-banded by mild trypsin digestion and Giemsa staining. R-bands (replication) and SCEs can be visualized by the incorporation of BrdU into replicating chromosomes followed by differential Giemsa staining. Accomplishment of these specific aims will provide evidence on the role of chromosomal changes involved in the development, long term existence, expression of the metastatic phenotype and developmental potential of Lucke' tumor and normal embryonic cells. Finally, this project will provide a significant opportunity for students to gain hands on experience in biomedical research and should act as a motivating factor encouraging their entry into a research oriented career.

fluorescence microscopy; gastrointestinal hormones; electrophysiology; cell biology; biomedical facility; alternatives to animals in research; fluorescent dye /probe; biological signal transduction; acidity /alkalinity; calcium flux; intracellular; peptide hormone; Xenopus oocyte; microinjections; voltage /patch clamp;

The new Image Analysis Core (IAC) of the Michigan Diabetes Research and Training Center (MDRTC) is a state-of-the-art facility, which was established at the University of Michigan in 1991. It is equipped with modern light and electron microscopic facilities which are interfaced with a powerful, computerized image analysis system. The Fluorescent Imaging Unit of the IAC is equipped with instruments for the visualization and quantification of various intracellular ions in living cells. The purpose of the IAC is to a) provide MDRTC investigators with morphometric analyses and quantitative cellular and subcellular localization of molecules and ions, using the techniques of quantitative histochemistry, in situ hybridization and fluorophotometric ion analysis, b) provide consultation to MDRTC investigators for the design and conduct of research projects using these techniques, c) initiate, implement and disseminate to MDRTC investigators new and innovative image analysis techniques to foster research on diabetes, metabolic and endocrine diseases, and d) demonstrate and teach the above techniques to MDRTC investigators, trainees and technicians. The major objective of the IAC will be to provide state-of-the-art quantitative morphological analyses using molecular probes, with quality and development having priority. The IAC is expected to enhance and stimulate diabetes and related endocrinology and metabolism research by providing scientific collaboration, expert assistance and special facilities that are essential for the research of Center investigators and trainees but that are beyond their individual capabilities.

9810524 Van BOCKSTAELE The brain nucleus locus coeruleus (LC) has been intensely investigated because it provides the neurotransmitter norepinephrine to all major levels of the brain and spinal cord. Neurons in the LC are the first norepinephrine- containing cell group to develop in the central nervous system. Numerous lines of research have shown that norepinephrine from the LC may be involved in regulating neuronal outgrowth in regions of the brain which receive norepinephrine innervation, such as cortical and limbic brain regions. Interestingly, the intrinsic physiological activity of LC neurons varies during development and may account for their involvement in these functions. Electrophysiological recordings from neonatal LC neurons in brain slices display oscillations in their membrane potentials which are synchronous throughout the entire nucleus. The frequency of oscillations increases from 0.3 Hz to 3 Hz until about three weeks of age when they disappear. The synchronous activity observed in neonatal LC neurons is thought to be the result of electrical coupling between LC cells. However, gap junction proteins, the anatomical substrate for electrical coupling, have not been demonstrated by electron microscopy nor has their existence been established at different periods of development in the LC. In addition, there have been no electrophysiological experiments that have examined the modulation of electrical coupling by compounds known to inhibit or stabilize gap junctions. The primary objective of our research is to unequivocally establish the existence of gap junction proteins that can couple LC neurons to each other or to glial cells at different stages of development. The methods include dual labeling immunocytochemistry at the electron microscopic level in young and adult rat brain using antibodies specific for the gap junction protein family, known as connexins, and for the norepinephrine synthesizing enzyme, tyrosine hydroxylase. Addition ally, electrophysiological recordings from individual LC neurons will be conducted in vitro and to test their responses to agents known to modulate gap junctions. Finally, individual LC neurons will be intracellularly filled and examined for dye coupling at both light and electron microscopic levels. Taken together, the anatomical and electrophysiological results obtained from this research will shed light on the functional architecture of LC in development and will thus lay the groundwork for future studies aimed at elucidating the role of the LC/norepinephrine system in regulating control of its postsynaptic targets.

DESCRIPTION (Adapted from application) This proposal requests support for the 12th International Symposium on Regulatory Peptides which will be used to assist new and junior investigators in participating in the conference. This symposium, which originated with a focus on gastrointestinal hormones but now includes coverage of other regulatory peptides and related molecules, is held biannually around the world and is the major conference at which work on regulatory peptides in the gastrointestinal tract, brain and other tissues is presented. These peptides are of major importance in regulating function of the gastrointestinal tract and understanding of their roles in disease is increasing. The conference will be held on Mackinac Island, Michigan, September 16-20, 1998, and attendance of approximately 400 scientists is expected. The conference will consist of nine plenary lectures, each coordinated with a session of 10 minute contributed talks on the areas of biochemistry and biosynthesis of regulatory peptides, integrative biology of gut regulatory peptides, regulation of growth and development, cytokines and the induction of apoptosis, G-protein coupled receptors, intracellular signal transduction cascades, transcriptional mechanisms of gene regulation, gaseous neurotransmitters (NO and CO), and clinical aspects of regulatory peptides. The housing of all attendees in a single hotel with common meals will also facilitate interaction between attendees as will unprogrammed time for adequate discussion in the afternoons. Evening sessions will include the Morton Grossman and Victor Mutt Lectures. To encourage attendance by new/junior investigators fellowships will be provided to 40 individuals (students, postdocs and beginning faculty). Twenty fellowships will be supported by the funds requested in this proposal.

Several major pancreatic disease including pancreatitis, cancer and cystic fibrosis involve altered cellular regulation. Secretion of digestive enzymes by pancreatic acinar cells is largely controlled by increases in intracellular Ca2+ and diacylglycerol which result from activation of phospholipase C. However, this mechanism can not explain all the effects of secretagogues and hormones on cell growth, protein synthesis and metabolism. Recently a number of novel protein kinase cascades have been elucidated that play important roles in growth, differentiation and gene expression of a variety of cells. We have shown in published and preliminary studies that CCK activates three mitogen activated protein kinase (MAPK) cascades in rat acini leading to activation of ERKs (p42 and p44 MAPK), Jun Kinase and p38/Reactivating Kinase. In addition, CCK activates a distinct pathway in acini leading to p70 S6 Kinase which is sensitive to rapamycin and wortmannin. The overall aim of this proposal is to understand how the novel kinase cascades are activated in acini, the specific stimuli which activate them, and some of their biological functions. Four specific aims include: 1) to determine the mechanism by which CCK and EGF activate the Ras-Raf-MEK-ERK cascade; the importance of the adapter proteins Sch and Grb2 as well as Ras will be evaluated. 2) To determine the tyrosine kinase activated by CCK which phosphorylates Shc. This will involve analysis of the activation of Src and Src family members and focal adhesion kinase (FAK). 3) To determine the activation of p38MAPK, its mechanism of activation, and its role in regulating phosphorylation of small heat shock protein, and 4) the mechanism of p70 S6K activation and its role in pancreatic acinar protein synthesis. The studies will involve immunoprecipitation of kinases, Western blotting and kinase assays using specific substrates. Selective pathway activation and inhibition using specific inhibitors and expression of dominant negative mutant proteins by adenoviral vectors will be used to evaluate pathways leading to biological effects including amylase secretion, growth and protein synthesis. While the work is aimed at understanding the pancreatic acinar cell, it will also have implications for the regulation of other gastrointestinal cell types.

Osteoporosis is the most common debilitating bone disease affecting 24 million people in the U.S. An estimated 50 percent of women over 45 and 90 percent of women over 75 years of age have osteoporosis, of which approximately half will suffer osteoporotic fractures. The estimated cost of osteoporosis and related fractures is 7-10 billion dollars per year. Bone resorption by osteoclasts is an energy intensive process, requiring ATP hydrolysis by an H+-ATPase to drive proton secretion. Age-dependent changes in bone mass are accompanied by age-dependent changes in other parameters, including glucose intolerance. Osteoclasts are glucose responsive cells that resorb bone in a glucose concentration dependent manner with a Km of 3 mM. The data presented here indicate that osteoclasts have glucose concentration dependent changes in 1) ATP synthesis, 2) phosphate uptake, 3) tyrosine phosphorylation, and 4) phosphatidylinositol 3-kinase activity. In view of the high metabolic demands of bone resorption, I hypothesize that glucose directly alters the activity of osteoclasts by modulating critical signal transduction processes in these cells. The proposed research will define the molecular mechanisms and metabolic requirements responsible for regulating these signal transduction processes. The Specific Aims are to: I: Characterize The Expression Of Glucose Transport Protein (S) In Osteoclasts. A. Determine the effect of bone attachment on GLUT2 expression in osteoclasts. B. Determine whether glucose transporters are translocated to the plasma membrane in response to bone attachment. II: Characterize Metabolic Elements That Govern Osteoclast Activity. A. Determine the rate limiting steps in glucose metabolism by osteoclasts. B. Characterize the glucose dependence of metabolic pathways. III: Identify Ket Signaling Pathways That Regulate Osteoclast Metabolism. A. Determine and characterize the role of glucose in osteoclastic phosphatidylinositol-dependent signaling pathways. B. Determine the role of glucose in modulating changes in tyrosine phosphorylation of specific proteins.

9972941
Milligan

In this research, the fundamental aspects of deformation instabilities in four classes of materials are studied to understand the similarities and differences in mechanical response and deformation mechanisms. The materials chosen are metallic glasses, multi-phase alloys containing a metallic glass matrix, metals with grain sizes between 30 nm and 3 microns, and amorphous polymers. Each of these materials exhibits a tension-compression asymmetry in the flow strength and deforms by shear banding on planes, which are not predicted by classical continuum mechanics. The understanding of these phenomena is incomplete, and there are large discrepancies in the literature concerning the sense of deviation of the shear band plane as well as whether the deviation and tension-compression asymmetry are due to hydrostatic pressure or normal stress effects. Further, the sense of the deviation of the shear band plane, which provides insight into the deformation mechanisms, is inconsistently reported in the literature, even for the same material. Performing mechanical tests on a single machine in a single laboratory should clarify these issues. Shear bands are characterized by advanced techniques, including atomic force microscopy and atomic resolution transmission electron microscopy aided by focussed ion beam thinning techniques. Studies of local chemistry changes in and around the shear bands are conducted with resolutions on the order of 1 nm. Mechanics analyses promote understanding of shear band widths, angles, spacing and statistical distributions, as well as effects of stress state and shear band configuration on the overall constitutive behavior. The grant is co-funded with the MPS Office of Multidisciplinary Activities and the Engineering CMS Mechanics and Materials Program.
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Bulk metallic glasses and bulk nanostructured metals are two classes of novel structural materials, which have attracted substantial interest. Bulk metallic glasses have found commercial applications in sports equipment and the continued development of nanostructured metals and composites might lead to commercialization of these materials. A common link between bulk metallic glasses and nanostructured metals is their mechanical behavior. Both materials shear band as the first and only mechanism of plastic deformation and exhibit no strain hardening. Similar behavior is observed for many amorphous polymers. This shear banding is of fundamental interest and of critical technological importance to the further development of these materials since shear banding is responsible for limited tensile ductility and anomalously low fatigue performance.
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9977462
Miskioglu
This is an equipment grant supported under the MRI Program. The instruments to be acquired are Nano Indenter XP and its related accessories which are able to deform very small volumes of material to characterize a material's mechanical properties. This instrument can measure the properties of extremely thin films and coatings, individual phases within a multi phase or composite material as well as other materials not well suited for conventional testing. In the dynamic nanoindentation option which uses the Continuous Stiffness Measurement (CMS) technique in conjunction with the Dynamic Contact Module (DCM), the system can provide a direct and accurate measurement of the contact stiffness at all depths. Material properties may thus be determined continuously. The Lateral Force Measurement (LFM) option provides an effective tool in characterizing scratch resistance in bulk materials and coatings, and adhesion and ode of failure in thin films. The instrument will be utilized in research related to micro-structural characterization of materials. These research activities address issues dealing with light weight composites with fine fibers and/or small particles, thin films, multi-phase polymeric systems, surface mechanical properties of silicon wafers and direct-write mesoscopic structures. In all these activities the nanoindentation system is expected to yield significant insight into the behavior of these materials.***

Chronic use of morphine results in tolerance to and dependence on the drug. One mechanism underlying cellular tolerance is an uncoupling of the opioid receptor from effects such that greater receptor occupancy is required to obtain a given response. This uncoupling mechanism has been studied using effectors that include; potassium and calcium conductances and the inhibition of adenylyl cyclase. A similar phenomenon is observed with acute opioid desensitization. Acute desensitization is thought to involve at least two linked processes, receptor uncoupling through an arresting dependent mechanism and removal of receptors from the plasma membrane through an internalization mechanism. Unlike more protracted forms of tolerance, acute desensitization is reversible within minutes and therefore easier to study in vitro. One goal of this proposal is to define the events that mediate the initiation and recovery from acute opioid desensitization. This goal has two parts, one is to characterize acute desensitization and determine the role of receptor internalization in that process. Given that receptor trafficking is an important form of receptor regulation, the second goal is to characterize how chronic morphine treatment alters desensitization and internalization of the opioid receptor. With a better understanding of the events that mediate desensitization and internalization the processes leading to long term tolerance may be more easily identified. The second goal is to identify post-synaptic cellular adaptations to chronic opioid treatment. These adaptations oppose the initial effect of opioid such that normal function is attained even in the continued presence of morphine. Thus adaptive mechanisms underlie an important form of tolerance. Two cell types will be examined the neurons in the locus coeruleus and interneurons of the VTA. There is an extensive knowledge of opioid actions in these areas, however, the identification and characterization of a post-synaptic adaptive mechanism studied in isolation has yet to be presented. Knowledge of alterations in regulation of ion channels during withdrawal form morphine may help in the development of more efficient protocols for the prevention of relapse to drug use.

DESCRIPTION (provided by applicant): The overall aim of this work is to understand how the growth of the adult pancreas is regulated to provide an adequate supply of digestive enzymes. We hypothesize that growth can occur by two mechanisms. The first occurs in response to diet and hormones and involves stimulation of differentiated acinar cells to enter the cell cycle and divide. The second form of growth that occurs following tissue injury such as pancreatitis involves dedifferentiation, cell division and dedifferentiation and will be referred to as regeneration. Our primary model for hormonal stimulation of growth involves feeding the synthetic protease inhibitor, camostat to mice to increase plasma CCK. We have shown this growth requires the calcium activated phosphates calcineurin and the mTOR pathway, and is accompanied by the activation of the MAP kinases, ERK and JNK, and the expression of a number of early response genes including multiple c-Jun and c-Fos family members. In Specific Aim 1 we will study the mechanisms mediating mitogenesis and growth of differentiated cells driven by chronic CCK elevation. a) We will identify the cell cycle regulators activated by CCK. b) We will study target genes regulated by NFATs (regulated by calcineurin) and AP-1 (activated by MAP kinases) with particular attention to regulation of cell-cycle regulating genes such as cyclin D1 and PCNA and to endogenous feedback inhibitors such as MClP1 for calcineurin. In Specific Aim 2 we will determine the mechanisms mediating regeneration after tissue injury. We will study regeneration following two models of pancreatitis induced by ethionine and caerulein over-stimulation. a) We will determine how the pattern of gene expression is altered in regeneration in the presence and absence of CCK using CCK deficient mice. b) We will determine the importance of calcineurin and mTOR for regeneration using both pharmacological inhibitors and gene targeted mice. In Specific Aim 3 we will determine the regulatory mechanisms for CCK and Growth Factor stimulated growth of acinar cells in primary monolayer culture which serves as a model for growth similar to regeneration and in which proteins and siRNA can be expressed with adenoviral vectors. We will first characterize the dedifferentiation and establish the acinar cell origin of dividing cells by lineage tracing. We will then evaluate the importance of different signaling pathways and cell cycle components with a focus on the key role of c-jun and AP1. Overall, the project will lead to better understanding of pancreatic growth and assist in designing approaches to regulating pancreatic growth in humans. PUBLIC HEALTH RELEVANCE: A number of pancreatic diseases including acute and chronic pancreatitis and pancreatic cancer lead to a loss of functioning pancreatic tissue. Knowledge from this project should allow designing approaches to assist the human pancreas to regenerate and ensure an adequate supply of pancreatic digestive enzymes.

The George Washington University Medical Center (GWUMC) and the Children's National Medical Center (CNMC), which is geographically separate but serves as the Department of Pediatrics of GWUMC will strengthen oversight of human subjects research at both institutions. A full time Research Compliance Educator (RCE) will be employed to coordinate and provide increased training for the investigators, staff and IRB members to ensure greater patient protections, promote patient's rights and understanding of the research they are part of, and train investigative teams in the ethical and scientific aspects of human subjects research. Educational initiatives will include (1) Protection of Human Subjects, (2) Institutional Review Board Policies and Procedures, (3) Use of the new on-line Biomedical Research and Information Network (BRAIN) for creating, routing, reviewing, approving and archiving human subject protocols, and for handling the submission, review and processing of adverse events, renewals, closures and suspensions, (4) Review of the effectiveness of educational programs and the development of new programs. Telecommunication equipment, which is available at GWUMC and CNMC, will be used to provide simultaneous educational programs at both institutions. Both institutions have agreed to continue the program after the completion of the grant award.

This award supports research to document the spatial and temporal modes of climate variation and land-cover change in eastern North America at millennial scales, assess the interactions between global forcings (orbital variations, trace gas concentrations) and regional controls (land-cover change, topography, lake effects) of climate and their relative effects on Holocene climate variability, and identify the critical mechanisms governing surface-atmosphere feedbacks.

This research will utilize a Regional Climate Model (RegCM2) in combination with a new generation of land-cover reconstructions to detail the effect of Holocene surface-atmosphere feedbacks on atmospheric processes. Model results will be evaluated against millennially resolved climate reconstructions for eastern North America based upon multiple networks of climate proxy records. The dense networks of proxy records and the large and regionally varied changes in Holocene temperature, aridity, and land cover make eastern North America an excellent case study for the interplay between forcings external to the climate system and internal feedbacks in temperate regions.

This research is important because climate variations during the Holocene were substantial, spatially variable, and spanned a wide range of frequencies and rates of change. Although these variations are in part forced by mechanisms external to the climate system, internal interactions among the atmosphere, oceans, and terrestrial biosphere are a critical source and modulator of climate variability. In particular, surface-atmosphere feedbacks are receiving increased scientific interest, due to recent demonstrations that such feedbacks exert a strong influence on Holocene and Recent climate processes at regional to global scales, with a magnitude comparable to orbital and greenhouse gas forcings.

In summary, this project will advance our understanding of Holocene climate variability and the interactions between climate and vegetation dynamics, enhance the scientific infrastructure, build partnerships among institutions, and help support a new generation of climate scientists. All model and data syntheses produced during the course of this work will be stored at the World Data Center upon publication.

DESCRIPTION (provided by applicant): The candidate's long-term career goal as a physician-scientist is to conduct investigator-initiated research into the molecular basis of the human immune response to viruses. Immediate goals include acquisition of skills in molecular virology and immunology studies in animals. These skills are critical to complement the candidate's extensive training in molecular human immunology acquired during previous studies of the human antibody response to respiratory syncytial virus (RSV). The experience gained during this project will prepare the candidate broadly to conduct unique studies of the immunobiology of early childhood pathogens. Human metapneumovirus (hMPV) is a newly described paramyxovirus that infects most humans by the age of five years. Our preliminary data indicate that hMPV is a major cause of lower respiratory tract infection (LRI) in children with a spectrum of illness and morbidity similar to that of RSV. This work proposes to define the antigenic determinants of protective immunity to hMPV, providing foundational new knowledge about the host immune response to this important emerging pathogen. The central hypothesis of this proposal is that the hMPV surface glycoproteins F (fusion) and G (attachment) are the major determinants of protective immunity and that protection is mediated by humoral antibody. Specific aim #1 is to investigate the antibody response to hMPV F and G proteins expressed in a novel vaccinia-virus based mammalian system. These recombinant antigens will be critical tools to evaluate the host immune response to hMPV. Specific aim #2 is to determine whether hMPV F and G surface proteins expressed by recombinant vaccinia virus are major protective antigens in a rodent model. These experiments will identify the determinants of protective immunity and define antigenic diversity between hMPV strains. Specific aim # 3 is to determine if humoral antibody alone is sufficient to mediate protection against recurrent LRI caused by hMPV. This work will provide valuable new data regarding the host immune response to an important respiratory pathogen of childhood that will be applicable to studies of human immunopathology associated with hMPV infection. The elucidation of the mechanism of protective immunity will also be important for future development of potential vaccines. The proposed work will provide the candidate skills and experience to be an outstanding physician-scientist in the field of human viral immunology.

The George Washington University Medical Center (GWUMC) and the Children's National Medical Center (CNMC), propose to continue the development of an educational program to strengthen oversight of human subjects research at both institutions under specific aim #1. As such, a full time Research Training Coordinator will be employed to coordinate and provide required training for investigators, administrative staff, students and IRB members to ensure greater patient protections, promote patient's rights and better understanding of the research they are part of. This training will be extended to investigative teams to strengthen their awareness of the ethical and scientific aspects of human subjects research. These educational initiatives pre-established under the previous RFA are critical to the ethical conduct of clinical research and will support Specific Aim #2, the accreditation of the IRB Program at GWU, at the conclusion of the award period.

[unreadable] DESCRIPTION (provided by applicant): Lower respiratory tract infection is a leading cause of morbidity and mortality in early childhood. These infections are primarily caused by viruses, such as respiratory syncytial virus (RSV), parainfluenzavirus (PIV) and influenza. Recently investigators in the Netherlands and Australia have described a new member of the Paramyxovirus family, tentatively named human Metapneumovirus (hMPV). The virus was associated with both upper and lower tract respiratory illness (LRI) but the epidemiology has not been studied. This study will investigate the incidence and clinical features of hMPV infection in children. The Vanderbilt Vaccine Clinic provides primary medical care to a large cohort of children who routinely have surveillance cultures of respiratory secretions and blood samples banked for serological studies. Hundreds of nasal wash and serum specimens are available for study, along with clinical data that has been prospectively collected in a blinded fashion by the study nurses. In small preliminary studies, we have used RT-PCR to identify hMPV in numerous frozen nasal wash specimens from children who previously tested negative for other known respiratory pathogens. The isolates share homology with the Netherlands strains, but show evidence of genetic diversity. Clinical symptoms associated with these infections were similar to RSV infection, characterized by fever, cough, wheezing and significant respiratory illness. A larger comprehensive study is needed. The specific aims of this study are to define the prevalence of hMPV infection in children, describe fully the spectrum of clinical illness and identify serologic markers of seroconversion and correlates of immunity. [unreadable] [unreadable] [unreadable]

DESCRIPTION (provided by applicant): This revised application requests continuing support for a training program which provides graduate students studying for the PhD degree with broad state-of-the-art training in Systems and Integrative Biology. Students will learn and apply tools and approaches of genomics, bioinformatics, proteomics as well as cell and molecular biology to the study of integrated systems and organismic biology. The program is centered in the Department of Molecular &Integrative Physiology because of its longstanding commitment to integrative biology but students will enter from an initial year in a combined gateway Program in Biomedical Sciences (PIBS) and can be enrolled in multiple PhD programs including Physiology, Pharmacology, Neuroscience, Bioinformatics and Human Genetics. Trainees will usually be supported for two years. The potential applicant pool is all students applying to PIBS and includes both students who come to Michigan to study integrative biology and those who enter undecided and are then drawn to integrative biology. The training includes formal coursework and seminars, teaching, and research experiences with faculty in a variety of departments at the University of Michigan. All students will take graduate level courses in Integrative Biology, Cell and Molecular function, Molecular Biology and Genetics, and Computational Biology. Additional courses are selected by the student in accordance with his/her interests. Students also participate in weekly student research seminars in their PhD program which they are expected to critically evaluate the presentations of their peers. This broad training is further reinforced by each student taking a written preliminary examination at the end of the second year of study that requires and understanding of function, from the cellular and molecular level to that of the organism including interactions between the environment and the organism. A unique aspect of the proposed training is that all students will have two dissertation co-chairs, rather than the more traditional single chair, at least one of whom will be schooled in the most recent techniques in cellular, molecular, or computational biology and one being an integrative biologist. The training faculty consists of 58 outstanding scientists all of whom are expert in either cellular and molecular or systems and integrative biology and committed to training individuals to answer questions of integrative physiological relevance. PUBLIC HEALTH RELEVANCE: This training grant will increase the number of scientists trained in systems and integrative biology. This area is important as it prepares scientists to participate in translational work on health and disease.

DESCRIPTION (provided by applicant): Synaptic transmission mediated by monoamines is terminated by cocaine-sensitive transporters. The overall hypothesis is that adaptations in monoamine release result from acute and repeated treatments with cocaine. The aims of this proposal are to identify and characterize synaptic adaptations in the Ventral Tegmental Area (VTA) that result from acute and repeated cocaine treatments. One aim of this proposal is to completely characterize an inhibitory postsynaptic potential caused by the synaptic release of dopamine in the VTA. The hypothesis is that acute and repeated administration of cocaine will change pre- and postsynaptic elements that control this synaptic response. By recording the effects of synaptically released dopamine on dopamine neurons this study will be able to identify adaptive mechanisms at both pre- and postsynaptic sites. A second aim is to identify the mechanism by which intracellular calcium regulates the activity of D2- dopamine receptors. When internal calcium is low, the activity of D2-dopamine receptors is high. The hypothesis is that postsynaptic interactions between calcium and D2 receptors will be sensitive to cocaine given both acutely and with repeated treatments. The significance of the proposed studies is two fold. First, the regulation of the firing of dopamine cells by endogenous dopamine through D2-autoreceptors has been known for decades. The proposed experiments will address cellular aspects of that regulation and the effects of acute and chronic cocaine that were not possible previously. Second, the results of this study on both the release of dopamine and sensitivity of D2-dopamine receptors will be a starting point for the understanding of the adaptive changes in the physiology of the dopamine system following cocaine treatment.

Geoinformatics: Collaborative Research - Late Neogene Terrestrial Ecosystem Database

Russell Graham, Pennsylvania State University, EAR-0622349
Eric Grimm, Illinois State Museum, EAR-0622289

Abstract
The "Late Neogene Terrestrial Ecosystem Database" will include fossil data for the past 5 million years (the Pliocene, Pleistocene, and Holocene epochs), the time during which modern species, including humans and modern ecosystems appeared. The purposes of the database are (1) to facilitate studies of ecosystem development and response to climate change, (2) to provide the historical context for understanding biodiversity dynamics, including genetic diversity, (3) to provide the data for climate-model validation, (4) to provide a safe, long-term, low-cost archive for a wide variety of paleobiological data, and (5) by to lower the overall cost of paleodata management. This database fulfills two critical needs: (1) it is an archive of invaluable paleobiological data and (2) it is essential infrastructure for paleoenvironmental research. This new database will integrate four existing databases into a single integrated database. The existing databases are the Global Pollen Database, the North American Plant Macrofossil Database, FAUNMAP (a mammalian database), and BEETLE, a fossil beetle database. These databases are currently in out-of-date PC database management systems, which are not suitable for Web servicing. They will be merged into a new database using state-of-the art software for Web servicing and using a completely new data model suitable for disparate types of paleobiological data. The databases that will be subsumed have become critical infrastructure for paleobiological research. They are used for almost every aspect of this research, from broad-scale synoptic studies to understanding the context of local site studies to using the database as a taxonomic standard. They are also widely used as teaching tools. Their integration will facilitate new lines of research and even more use of these data.

The Cell Biology and Cell Imaging Core is a new Core that has evolved from aspects of two previous[unreadable] cores, the Cell Biology Core which focused on live cell imaging, and the Histochemistry/Morphology[unreadable] Core which focused on cell structure and the localization of biological molecules in fixed cells. The Cell[unreadable] Imaging Core is designed to provide information on the localization of molecules in live and fixed cells[unreadable] and their changes over time and in response to physiological and pathophysiological perturbation. It is[unreadable] centered around microscopic imaging and quantitative analysis of digital information obtained primarily[unreadable] by last scanning confocal microscopy (LSCM) and multiwavelength fluorescence imaging although[unreadable] traditional electron microscopy of fixed and embedded specimens is also available for fine structure[unreadable] analysis.

In the eastern United States, forest plant communities that were present 17,000 to 12,000 years ago are no longer represented on the landscape. This project will determine whether these historic assemblages of vegetation arose from: 1) unusually high temperature seasonality, 2) low atmospheric carbon dioxide concentrations, and/or 3) herbivory by now-extinct ''megafauna'' (e.g. mammoths, mastodons). This project integrates paleoecological data and mechanistic vegetation models. Specifically, this project will study lake-sediment cores from seven mid-continental sites from Tennessee to Minnesota and model climatic and carbon dioxide controls on vegetation composition. Sediments will be analyzed to infer ecological histories of vegetation, fire, and megafaunal population density and determine the timing of key ecological events.

This study of past environmental change has direct implications for current climate-change concerns. In particular, for planning purposes, there is an urgent need for accurate ecological forecasts, but the robustness of current ecological models, when applied to environmental conditions outside the range of modern observations, is poorly understood. Moreover, climates at the end of the 21st century will likely include combinations of temperature and precipitation unlike any observed today. The well-documented anomalous late-glacial climates and ecological communities are a good testing ground for evaluating the adequacy of current models of ecological response to complex environmental change. This work will support student training and outreach activities for high school teachers and students.

[unreadable] DESCRIPTION (provided by applicant): A critical number of investigators at the Kimmel Cancer Center of Thomas Jefferson University have a number of pending projects that require a quantitative method for measuring the quaternary structure of biological samples and their association constants. To this end, we propose to acquire the Beckman Optima XL-I analytical ultracentrifuge. This instrument is well suited for these projects and significantly extends the capabilities of our molecular interaction core facility. Specific areas that we will immediately address using this instrumentation are: 1) Thermodynamic characterization of multimeric dynein and dynactin complexes in relation to the regulation of dynein activity and cargo binding; 2) To study the oligomerization state and hydrodynamic properties of peptide segments derived from viral glycoproteins involved in membrane fusion; 3) tRNA-protein complexes and the mechanism of decoding genetic information; 4) Characterization of Protein-Protein Interaction Antagonists; 5) Characterize conformational changes induced by activation of arrestins, a family of proteins that regulate G protein-coupled receptor signaling; 6) Characterization of the equilibrium binding constants and the stochiometry of the ectodomains of the TCR, the pMHC and the CD8. In addition, the instrument will enhance two additional core facilities (X-ray and small molecule screening facilities) as well as graduate education (PR613, a graduate course on structural biology, is structured such that students apply each technique taught to an actual sample). The analytical ultracentrifuge of choice is the Beckman Optima XL-I. This instrument permits detection using either interference optics or UV/visible optics. The interference mode is sensitive to the refractive index, and thus, does not require the analyte to have a natural chromaphor. Thus, this will permit measurement of samples with low molecular extinction coefficients, small molecule antagonist and membrane bound proteins. The interference optics also provides rapid data collection useful for producing high quality data for sedmentation velocity analysis. On the other hand, the ability to measure absorbance over a large range (190 to 800 nM) allows simultaneous measurement of individual components in a multicomponent system. [unreadable] [unreadable] [unreadable]

[unreadable] DESCRIPTION (provided by applicant): HIV continues to spread among Black men who have sex with men (MSM), but few interventions target high- risk Black men who have sex with men and women (MSM/W). Black MSM/W with histories of childhood sexual abuse (CSA) may be an especially vulnerable population for: a) high-risk sexual behaviors; b) negative psychological sequelae (e.g., depressive or posttraumatic stress disorder (PTSD) symptoms); and c) neurobiological abnormalities in cardiovascular, neuroendocrine and/or immune systems (e.g. cortisol and norepinephrine), and HIV/AIDS progression. This 3-year R34 pilot study, An HIV Intervention for Black Men at Risk, builds on the evidenced-base NIMH-funded Women's Health Intervention and a state funded HIV risk reduction pilot, The Men's Sexual Risk Reduction Project, for HIV positive gay-identifying African American and Latino MSM with histories of sexual abuse. This study will consist of three phases: Phase I. The Formative Phase will include three 90-minute focus groups to identify feasibility issues and factors pertinent to recruiting and conducting a psychosocial intervention such as perceptions of masculinity and gender roles. Two groups will include HIV positive Black MSM/W with histories of CSA and the third group with community leaders that work with non-gay identifying Black MSM/W. In Phase II-The Pilot, we will modify the existing manualized Men's Sexual Risk Reduction intervention and pilot it on 8-10 non-gay identifying HIV positive Black MSM/W with histories of CSA. In Phase Three-The Intervention, guided by the Social Learning Theory, the Ecological Model, and the concept of allostatic load, a composite of the cumulative effects of stress on biological systems including psychoneuroimmonologic markers, we will test the 6-session Enhanced Sexual Health Intervention for Men (ES-HIM) on 88 non-gay identifying HIV positive Black MSM/W with histories of CSA. The outcomes are to reduce high-risk sexual behavior (i.e., unprotected anal and vaginal sex) and negative psychological symptoms of depression and PTSD. Links between these outcomes and biomarkers of stress will be explored. Randomization to either the ES-HIM or a health promotion (HP) control group will occur with participants assessed at baseline, post, 3- and 6-months. A full spectrum of univariate and multivariate approaches will be used to evaluate the efficacy of randomization at baseline, to test for group differences at baseline as a function of CSA severity, as well as to test for treatment effect differences over time on the primary outcomes of interest. The findings from this pilot study will be used in the application for a larger NIMH R01 intervention. [unreadable] [unreadable] The Enhanced Sexual Health Intervention for Men (ES-HIM) is a biopsychosocial HIV risk reduction intervention. Its aims are to decrease high-risk sexual behaviors and depressive and PTSD symptoms among HIV-positive non-gay identifying Black men who have sex with men and women and who have a history of childhood sexual abuse. Besides improving mental health and helping to decrease the risk of HIV re-infection and transmission among these men and their male and female partners, ES-HIM explores the relationship between these outcomes and biomarkers of stress. Understanding these associations may be important in improving mental and physical health of this vulnerable population of men. [unreadable] [unreadable] [unreadable] [unreadable] [unreadable] [unreadable]

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[unreadable] DESCRIPTION (provided by applicant): Human coronaviruses (HCoV) were initially identified as major causes of acute respiratory tract disease in the 1960's. The epidemiology of two HCoV, OC43 and 229E, was established using only serological methods, due to difficulty culturing the viruses. Three new HCoV have been described in the last three years [1-4]: severe acute respiratory syndrome coronavirus (SARS-CoV) [4], human coronavirus Netherlands (HCoV-NL) [1, 2], and human coronavirus Hong Kong (CoV-HKU1) [3]. Each of these new viruses has been described in association with lower respiratory tract illness (LRI). The discovery of these viruses has renewed interest into the previously known HCoV OC43 and 229E which were known to cause upper respiratory tract illness (URI). HCoV OC43 and 229E have also been associated with LRI like the newly discovered coronaviruses. However, there are no published large-scale epidemiologic studies of OC43 and 229E using highly sensitive molecular diagnostic methods. The objectives of this study are to define the prevalence and clinical illnesses associated with the new human coronavirus HCoV-NL and the prototypic HCoV, OC43 and 229E, over a 20-year period in a cohort of over 2,000 previously healthy outpatient children. We also will determine the genetic variability of these viruses by amplifying and sequencing the HCoV spike protein gene. The Vanderbilt Vaccine Clinic (WC) clinical database and sample archive provide us with the tools to accomplish these goals. The WC provided comprehensive medical care to a large cohort of outpatient children who routinely had surveillance cultures of respiratory secretions and serum samples obtained, thus thousands of nasal wash and serum specimens are available for study, along with robust, prospectively collected clinical data. This provides a unique opportunity to investigate the importance of emerging human pathogens. Our preliminary data using samples from the WC indicate that coronaviruses are a significant cause of respiratory tract infection in children with a spectrum of illness and morbidity. [unreadable] [unreadable] [unreadable] [unreadable]

[unreadable] DESCRIPTION (provided by applicant): The central objective of this proposal is to determine mechanisms of entry for human metapneumovirus (hMPV), a recently described paramyxovirus that is a major cause of lower respiratory tract infection in infants and children worldwide. Understanding the virus-cell interactions that lead to docking and internalization of hMPV is critical to elucidating mechanisms of pathogenesis. Host cell receptors are important determinants of virus tropism and can evoke signaling pathways that lead to virus internalization or prime the cell for viral replication. Furthermore, an understanding of virus-receptor interactions may lead to development of targeted antiviral compounds, such as pleconaril, which blocks picornavirus binding to its receptor. Several pathogenic human viruses utilize RGD-binding integrins as receptors or co-receptors, including adenovirus, Coxsackievirus, parechovirus, rotavirus and human herpes virus-8. Our preliminary data suggest that RGD- binding integrins serve as receptors for hMPV and that the fusion (F) protein mediates this interaction. In Aim 1, we will define mechanisms by which integrins mediate hMPV cell entry. hMPV infectivity requires 1V21 integrin, and this interaction is mediated by hMPV F protein. We hypothesize that hMPV F-integrin interaction is not a simple binding event, but that integrin ligation triggers fusion. Wild type and mutant recombinant F protein will be tested for the capacity to bind 1V21 integrin in flow cytometry and surface plasmon resonance assays. Mutant viruses with alterations in integrin-binding motifs will be generated by reverse genetics and tested for the capacity to bind 1V21 integrin and infect cells. The contribution of integrin binding to membrane fusion will be determined using a fusion assay based on a stably transfected cell line expressing wild type or RGD-mutant hMPV F. These experiments will determine the role of 1V21 integrin in hMPV cell attachment, fusion and entry. In Aim 2, we will determine whether integrin engagement by hMPV F triggers intracellular signaling. Integrins mediate both outside-in and inside-out signaling. These experiments will determine whether integrin activation is required for hMPV fusion and cell entry and whether downstream signaling is mediated by hMPV F-integrin binding. We will use experimental conditions that promote or hinder integrin activation to determine whether integrin activation is required for F binding, for fusion and for cell entry. These three steps in hMPV entry will be determined using a recombinant F protein binding flow cytometry assay, a cell-based fusion assay, and hMPV virus infectivity assay. Separate experiments will be performed to determine the effect of F binding on downstream signaling such as focal adhesion kinase (FAK) phosphorylation and the recruitment of second mediators to 1V21 integrins. Inhibitors of downstream integrin signaling, immunoprecipitation and siRNA techniques will be used to determine whether integrin-mediated signaling is required for hMPV entry or replication. These experiments will contribute broadly to an understanding of how integrin signaling properties are utilized by pathogenic viruses. PUBLIC HEALTH RELEVANCE: Human metapneumovirus (hMPV) is an emerging virus that was discovered in 2001. hMPV is a major cause of respiratory tract disease worldwide, and leads to many hospitalizations of infants and children. The purpose of this proposal is to determine the protein (or "receptor") on human cells that hMPV attaches to in order to enter and infect the cell. These studies will increase understanding of how hMPV causes disease in humans, and may help develop targets for antiviral medicines and vaccines. [unreadable] [unreadable] [unreadable] [unreadable] [unreadable] [unreadable] [unreadable]

DESCRIPTION (provided by applicant): The development of tolerance to opioids limits their effectiveness for the treatment of pain. A wide variety of opioids that are commonly used in the clinic for the treatment of pain and drug addiction have dramatically different pharmacological properties. One primary difference between agonists is the ability of different agonist to cause varying amounts of desensitization and internalization of mu opioid receptors. Agonists fall into three major groups, those that induce both desensitization and internalization, those that induce desensitization but not internalization and those that are ineffective at both. The role that desensitization and internalization have in the development to tolerance and dependence to opioids has been a controversial subject that has been studied in a variety of model systems. This exploratory proposal will develop the method of Flurescence Correlation Spectroscopy to investigate the mechanism that accounts for the varying actions of different opioid agonists on the mu opioid receptor. This is a relatively new method that uses optical measurements of the mobility of fluorescent molecules within a very small volume. These measurements are made at the plasma membrane, within intracellular compartments in various parts of the cell, including the cell body, dendrites, axons and terminals. The mobility of molecular is directly related to the molecular interactions such that agonist/receptor and receptor/effector associations will be identified. This study will use agonists that are fluorescent (aim 1) and fluorencescently tagged mu-opioid receptors (aim 2) in a model system, HEK293 cells that stably express epitope-tagged mu-opioid receptors. Electrophysiological, biochemical and imaging methods have been used to characterize many of the steps in opioid-receptor dependent signaling. This exploratory proposal will introduce a new way to investigate the mechanisms that underlie the different pharmacological profiles of opioid agonists. In the 2 years afforded this proposal, this method will be developed using a very well characterized model, the HEK293 cells. Once this method is established, the ultimate goal of applying it to study of primary neurons will be pursued. By gaining knowledge of the processes that are selectively affected by some agonists and not others, the mechanisms underlying the development of tolerance and dependence may be identified and applied to more effective treatment of chronic pain and addiction.

DESCRIPTION (provided by applicant): The major objectives of the proposed research in this application from a new investigator are to develop candidate vaccines for human metapneumovirus (HMPV), identify viral determinants of virulence and define genetic diversity of HMPV over time. HMPV is a recently discovered paramyxovirus that is a major cause of lower respiratory tract disease in infants and children worldwide, causing hospitalization and fatal infections. HMPV is also associated with asthma exacerbations requiring hospitalization and causes severe disease in persons with underlying conditions such as prematurity, cardiopulmonary disease, and cancer. This work proposes to develop live attenuated vaccines against HMPV and define viral determinants of virulence. We propose three specific aims: Specific Aim 1. Develop cell culture-passaged strains of HMPV and determine their pathogenicity and immunogenicity in animal models. Specific Aim 2. Define viral factors that contribute to virulence and pathogenicity. Specific Aim 3. Determine the genetic diversity and evolution of HMPV over time. This research will provide potential vaccine candidates for HMPV and provide new knowledge about pathogenesis of this important human pathogen. The proposed experiments use a combination of animal studies, classical virology techniques, cellular immunology tools, and high-throughput sequencing to define HMPV virulence factors and identify attenuating mutations. The findings will be useful for the design of reverse- engineered HMPV vaccines. Leading vaccine candidates for the related respiratory syncytial virus use reverse genetics to incorporate attenuating mutations identified by similar methods, as well as complete gene deletions. The use of high-throughput sequencing methods will provide novel information about the genetic diversity of HMPV over time. This information is essential for designing vaccines or prophylactic prevention strategies. We hypothesize that, like other paramyxoviruses, HMPV will not exhibit substantial genetic or antigenic drift over time. However, these data are not known and it is possible that HMPV would demonstrate continued mutation similar to influenza virus, requiring a new vaccine each year. HMPV has a major impact on human health and safe, effective vaccines could decrease the burden of disease associated with this novel pathogen. PUBLIC HEALTH RELEVANCE: Human metapneumovirus (hMPV) is an emerging virus that was discovered in 2001. hMPV is a major cause of respiratory tract disease worldwide, and leads to many hospitalizations of infants and children. The purpose of this proposal is to develop candidate vaccines for hMPV. These studies will increase understanding of how hMPV causes disease in humans, and may help develop targets for antiviral medicines and vaccines.

NON-TECHNICAL ABSTRACT
Silicon-based electronic devices are the main component in virtually all microelectronic applications, e.g., computers and computer chips that are everywhere in cars, appliances, etc. For high-power and high-temperature uses, however, e.g., on-engine chips, power grid controls, etc. Si-based electronics are either inefficient or not usable at all. Silicon carbide is the most promising alternative, but, despite major advances in the last decade, including breakthroughs by the present team, difficult technical problems remain to be resolved. The proposed research addresses these issues with a mix of experimental and theoretical techniques. At the heart of the difficulties is the interface between silicon carbide and its native oxide, namely silicon dioxide, and the impact of the oxidation process on the underlying crystalline material. The expected results will be relevant to the broader field of the oxidation of diverse materials. Educational outreach projects will highlight to high school teachers and students the special needs of high-power, high-temperature electronics and the impact of research advances in important applications.

TECHNICAL ABSTRACT
Silicon carbide is a promising alternative to Si for high-temperature, high-power electronics because of its larger energy gap and heat-conduction coefficient, but also because its native oxide is silicon dioxide. The SiC/SiO2 interface, however, is more complex that the Si/SiO2 interface, which is at the heart of Si-based electronics. The main problem is that oxidation releases C atoms, some of which are stuck at the interface as defects. Despite major advances in passivating defects at the SiC/SiO2 by N and H, including breakthroughs by the present team, electron mobility remains lower than desirable for applications. Recent evidence points to subtle changes in the underlying SiC substrate. The proposed research will combine state-of-the-art microscopy, electrical measurements, and theory to elucidate the oxidation process at the atomic scale, identify the origins of undesirable effects and defects, and identify new design specifications to improve carrier mobilities. Extensive education outreach will make advances accessible to high schools and the community.

Floral and Faunal Community Responses to Late-Quaternary Climate Change

John Williams, Stephen Jackson, Eric Grimm and Russell Graham


The PIs propose a study of the responses of species and communities to
late- Quaternary environmental variations and, in particular, the environmental drivers of species turnover, community assembly and disassembly, and the formation of no-analog communities. Their research will demonstrate the power of NEOTOMA, a new community paleoecological database that reduces informatics costs and removes barriers to interdisciplinary collaboration by storing in a single database most of the major late-Neogene paleoecological databases. The proposed research will be the first-ever synoptic analyses of late-Quaternary community dynamics in North America that directly integrate fossil pollen, plant-macrofossil, and faunal records. The PIs propose improving the resolution of the NEOTOMA database by obtaining 100 new AMS radiocarbon dates for key vertebrate fossil localities, adding recent high-quality records to NEOTOMA, revising existing chronologies in NEOTOMA in light of this new data, and mapping all data for 15 time-windows covering the last 21,000 years. These time windows are more finely subdivided than the original FAUNMAP synthesis, and will permit study of species responses to the rapid climate changes of the last deglaciation. The PIs will test such hypotheses by comparing the distributions of the floral and faunal no-analog communities to each other and to the distributions of no-analog climates. They will create plant based and animal based biome maps to see whether such maps are complementary or in contradiction. The PIs will further conduct generalized dissimilarity modeling to reveal patterns of species turnover, along environmental gradients, in both space and time.

This project will help support the continued development of the field of biogeography, an interdisciplinary study that integrates topics of importance in evolutionary biology, paleo-biology, conservation biology, and ecology. It will accomplish this through funding the travel costs of 40 US graduate students and post-docs to attend the annual meeting of the International Biogeography Society (IBS) in Merida, Mexico, in January 2009. The conference is organized to provide young scientists with networking and educational opportunities that will provide many benefits of broader impact to their professional development, as well as their intellectual development. The IBS meeting emphasizes interaction among scientists from diverse countries, institutions, and career stages, and is designed to highlight and explore the frontiers of the field, fostering the intellectual development of scientists and progress of the field. Additionally, this project significantly enhances international contacts between U.S. graduate students, foreign students, and other scientists of all career stages.

DESCRIPTION (provided by applicant): Microtubules represent one of the three essential cytoskeleton types in cells. Important for a variety of physiological functions, encompassing cell migration, mitosis, neuronal differentiation and transport of cargo, microtubule-associated motor proteins have been implicated in numerous diseases, ranging from motor neuron and degenerative disorders, to neoplasia and viral infections. Microtubule-binding CAP-Gly domains are conserved in organisms from human to yeast, play central roles in many proteins, and their mutations lead to various disorders. CAP-Gly domain of the p150glued subunit of dynactin interacts with microtubules, and its mutations are associated with several motor neuron disorders. The atomic-level structure and dynamics of CAP-Gly/microtubule assemblies are not known because of their inherent insolubility and lack of long-range order. Lack of such insight hampers further research and impedes design of effective therapies against diseases associated with cytoskeleton dysfunction. Our long-term goal is to understand the structural and dynamic basis of cargo transport regulation along microtubules by microtubule-associated proteins, in healthy and disease states. The objectives of this application are to determine three-dimensional structures and dynamics of CAP-Gly domain of dynactin and of its macromolecular assemblies with the microtubules and with EB1 protein. We will employ multidimensional high-resolution magic angle spinning solid-state NMR methods in conjunction with biophysical and biochemical techniques. In the specific aims designed to accomplish the objectives of this application, we will: 1) determine the structure of CAP-Gly alone and CAP-Gly assembled on the microtubule, and identify the CAP-Gly/microtubule interface at atomic resolution; 2) characterize the energetics and dynamics of the CAP-Gly/microtubule interaction; 3) characterize the dynamics of CAP-Gly mutants related to neurological pathologies; 4) characterize biochemically and structurally the regulation of the CAP- Gly/EB1/microtubule interaction. The proposed work has important implications for human health as it will shed light on the structure of CAP-Gly:microtubule complexes that are not amenable to structural characterization by X-ray crystallography or solution NMR spectroscopy, and will enable structural characterization of macromolecular assemblies consisting of microtubule-associated proteins in complexes with microtubules.

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. This project will focus on rational design to modulate antibody affinity.

DESCRIPTION (provided by applicant): In recent years monoclonal antibodies (mAbs) recognizing tumor-associated antigens have increasingly been integrated into the clinical management of neoplasia and this trend is expected to continue. Prominent targets for antibody-based therapies are ErbB family members, including the epidermal growth factor receptor (EGFR/ErbB1) itself and ErbB2. EGFR-specific antibodies currently in clinical use typically inhibit EGFR-dependent signal transduction and induce antibody-dependent cell-mediated cytotoxicity. Administration of these agents to tumor patients is, however, associated with dose-limiting adverse side effects due to inhibition of EGFR signaling in normal tissues, specifically skin and the gastrointestinal tract. It is, therefore, highly desirable to develop antibody-based therapeutics that specifically recognize their target antigen in the tumor environment but not on normal tissues. Here we propose a novel approach to achieve this goal. It is based on the construction of antibody derivatives in which antigen recognition sites are reversibly masked by antigen fragments. Association of the antigen fragment with the mAb CDRs is expected to occlude the antigen recognition site of the fusion protein and, thus, reduce normal tissue reactivity. To restore high affinity antigen- antibody interaction at tumor sites, a proteolytic cleavage site is engineered into the linker between the blocking moiety and the antibody fragment. This site is designed to be susceptible to proteases with high activity in tumor tissues (i.e. matrix metalloproteases (MMPs)) but little or no activity in normal tissues including peripheral blood under homeostatic conditions. Upon cleavage at tumor sites, the 'unmasked'antibody is predicted to partition to the tumor-associated antigen due to mass action principles. The proposed work will test biological properties of antibody derivatives based on the EGFR antagonistic mAbs C225 and 425 that recognize distinct epitopes on the extracellular domain of the human EGFR (EGFRdIII). In preliminary work dual (Ab-Ag)2 complexes were made whereby C225 and 425 scFvs were covalently linked to EGFRdIII sequences engineered to encourage dimerization and reciprocal occlusion (crossmasking) of the antigen recognition sites of each other. Upon MMP9 treatment these complexes were shown to dissociate followed by markedly enhanced binding to purified EGFR and to tumor cells expressing EGFR. These promising results form the basis for the reformatting and in vitro testing of crossmasked Abs in an IgG-like format (Specific Aim 1) and the determination of tumor homing and growth inhibition human tumor xenotransplants in mice by such constructs in vivo (Specific Aim 2). In future work, the utility of these reagents as diagnostic agents (imaging) will be further tested in appropriate animal models, i.e. human tumor xenotransplants in mice. PUBLIC HEALTH RELEVANCE: In recent years monoclonal antibodies (mAbs) have been used increasingly in the clinical management of certain forms of cancer and select other diseases. Most mAbs currently used in cancer therapy recognize tumor-associated antigens, which are also present on normal tissues. This circumstance leads to adverse side effects that limit the application and efficacy of these agents in patients. This proposal describes a novel concept to engineer antibody derivatives that overcome these limitations. To this end, we will focus on molecular modification of two EGFR antagonistic monoclonal antibodies currently in clinical use or clinical trials, i.e. C225 and 425, respectively. If successful, the concept to be tested here may be applicable to a wide range of mAbs currently in clinical use or development.

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. Primary support for the subproject and the subproject's principal investigator may have been provided by other sources, including other NIH sources. The Total Cost listed for the subproject likely represents the estimated amount of Center infrastructure utilized by the subproject, not direct funding provided by the NCRR grant to the subproject or subproject staff. Microwave assisted organic synthesis (MAOS) is being employed to synthesize cytotoxic arylphosphonium salts (APS). APS exhibit structure-activity relationships as antibiotics, in DNA binding, enzyme inhibition and against malignant cells. They cross cell membranes and accumulate in mitochondria of malignant cells. They deliver DNA alkylating agents into cancer cells, inhibit acetylcholinesterase, bovine serum amine oxidase, protein kinase C and HIV integrase. Among possible consequences are disruption of metabolism and cell division from blocked enzymes and interference with replication. A report of the latter with student co-authors has been published. APS effects in DNA melting, electrophoresis, and isothermal titration calorimetry are being studied. A student co-authored article on in silico APS-estrogen-receptor binding has been published. We will do further experiments at RIC and Brown EPSCoR on toxicity mechanisms of APS. We have synthesized new APS-fluorescent conjugates and observed their uptake by live cells. We have prepared APS polymers and graft polymers that are antibiotic. AutoDock, HyperChem and eHitsLightning will and are being used to calculate APS-DNA interactions. Polymeric APS will be tested further for antibacterial activity for potential use in medical plastics. New APS will be screened for DNA-replication toxicity. A crystallization project to make complexes for X-ray structure determination is underway and crystals have been isolated from DNA-APS hanging-drop experiments. Solid-state polypeptide synthesis will provide polypeptides for conjugates to APS and for DNA complexing studies. Triphenyltriazoles will be synthesized and incorporated into polypeptide co-polymers to make antibiotic and/or biodegradable macromolecules.

Neotoma is a multiproxy relational database that includes fossil data for the past 5 million years (the Pliocene and Quaternary). It is a community database that provides the underlying cyberinfrastructure for a variety of disciplinary database projects. The Neotoma project is an international collaborative effort among individuals from 23 institutions, including domain scientists representing a spectrum of Pliocene-Quaternary fossil data types, as well as experts in information technology (IT). This project builds on the initial development of the Neotoma database, which was funded in 2008 with a 2-year grant from the NSF Geoinformatics program. Neotoma involves domain scientists representing a wide spectrum of fossil groups from the Pliocene and Quaternary teamed with information technology (IT) professionals from the Center for Environmental Informatics (CEI) at Penn State, where the database is hosted. A workshop was held at the National Academy of Sciences in Washington DC on 31 January-2 February 2009 to plan for the future development of Neotoma. This project is the outgrowth of the DC workshop.

During a 2-year development project, the database structure of Neotoma was designed and implemented with the import of vertebrate faunal data from the FAUNMAP database and pollen data from the Global Pollen Database. The North American Plant Macrofossil Database was updated, and data are now ready for import. The Neotoma website was launched, and the Neotoma explorer provides a map-based interface to the data.

During the next phase ( this project) several new constituent database cooperatives from North America, Europe, Africa, and Asia will join with Neotoma, and several new data types will be ingested, including diatoms, ostracodes, insects, and testate amoebae. Neotoma will be exposed to outside developers via Application Programming Interfaces (APIs) and a Software Development Kit (SDK), whereby both desktop and webbrowser- based applications can acquire data from Neotoma over the Internet. Data-steward tools will be developed that will allow stewards to upload and update data remotely. This project establishes working groups for various database activities, including development of software tools, development of age models, integration of various fossil types from packrat middens held in different databases, and organization of a new peatlands constituent database. Two database-wide workshops will be held for coordination, planning, and conceptual development.

DESCRIPTION (provided by applicant): Dopamine plays a role in multiple physiological processes ranging from movement and reward to the regulation of hormonal balance. Dopamine receptor agonists or antagonists are used for the treatment of diseases such as schizophrenia, depression, attention deficit disorder, and Parkinson's disease. Recent work also indicates that the altered regulation of dopamine release induced by many drugs of abuse plays a critical role in early processes linked to early aspects of addiction. This there is a considerable understanding of the physiological role of dopamine in the central nervous system. There is likewise an enormous literature on molecular and biochemical aspects of dopamine signaling. Studies range from, the expression of the multiple is forms of dopamine receptors and second messenger cascades in cell lines to, biochemical and electrophysiological examination of the actions of dopamine and receptor agonists on neurons in various parts of the central nervous system. Thus the signaling cascades that are activated by dopamine receptors are well characterized. In spite of the wealth of knowledge of the dopamine system, little has been done on the actions of synaptically released dopamine at the cellular level. This hole in knowledge is not for the lack of effort, but lies in the fact that robust physiological detectors linked to dopamine synapses have been difficult to find in the major projection areas. This proposal uses recordings from dopamine neurons in brain slices from mouse to (1) define the inhibitory synaptic potential (IPSP) that results from the dendritic release of dopamine, (2) identify the presynaptic mechanism(s) and sites of dendritic dopamine release and (3) determine how the dendritic release of dopamine is altered following the induction of excitatory synaptic plasticity resulting from the treatment of animals with cocaine. The robust connection between the dendritic release of dopamine and the activation of an IPSC remains the best and probably only site at which dopamine transmission has been directly examined. The results of this study will therefore form a connection between knowledge at molecular and systems levels. The characterization of the synaptic mechanisms that determine rise and fall of extracellular dopamine in combination with a physiological response, will add considerably to the decades long search for an understanding of the role of dopamine in health and disease. PUBLIC HEALTH RELEVANCE: Dopamine agonists and antagonists are used clinically for the treatment of a number of diseases pointing out the importance of a complete understanding of the cellular and synaptic processes mediated by this important transmitter. The results from this study will define the cellular interactions between endogenously released dopamine and the receptors that detect the release of dopamine. From this work knowledge of dopamine-dependent transmission in many areas of the brain will be facilitated and lead to a better understanding of the disruptions in transmission in many diseases including addiction.

DESCRIPTION (provided by applicant): Human metapneumovirus (hMPV) is a recently discovered paramyxovirus that is a major cause of lower respiratory tract infection. hMPV causes repeat infections in children and adults and severe disease in older patients and individuals with underlying medical conditions. Mechanisms of hMPV immunity and pathogenesis are poorly understood and there are no licensed vaccines or therapeutics. The limited information available indicates that there are fundamental differences between hMPV and other paramyxoviruses, including respiratory syncytial virus. Accordingly, the central objective of the proposed research is to elucidate mechanisms of immunity to hMPV and define the contribution of host immune responses to disease. We propose three complementary specific aims to elucidate mechanisms of hMPV immunity and pathogenesis. In Specific Aim 1, the role of antibody in protection and disease will be defined. B cell-deficient mice will be infected with hMPV and viral growth and lung pathology assessed following secondary infection. Passive serum transfer experiments will determine whether antibody is required for protection. The role of antibody- dependent cellular cytotoxicity in immunity and disease will be determined using NK cell-deficient mice. In Specific Aim 2, the capacity of hMPV fusion (F) protein to mediate protection against reinfection will be determined. Mice will be immunized with F protein and challenged with engineered viruses encoding divergent F proteins. Immunized and challenged mice will be evaluated for respiratory disease and aberrant immune responses. These experiments will test the hypotheses that hMPV F protein vaccines will not be associated with enhanced disease and that hMPV F protein is capable of inducing broadly protective immunity. In Specific Aim 3, the contribution of T cells to hMPV immunity and pathogenesis will be defined. Naive T cell-deficient mice will be infected with hMPV, and viral replication and lung pathology will be assessed. T-cell-deficient mice will be inoculated with hMPV, allowed to clear infection, and rechallenged to determine the contribution of T cells to protection against secondary infection and lung pathology. CTL epitope vaccination and adoptive transfer of cloned MHC class I-restricted epitope-specific CD8+ T cell lines will be used to determine whether virus-specific CD8+ T cells contribute to pathology. Results of the experiments proposed in this application will reveal mechanisms by which host immunity can lead to both resolution of infection and immunopathogenesis. These findings will advance knowledge of hMPV immunopathology, guide the development of hMPV therapeutics and vaccines, and have broad applicability to other respiratory viruses.

DESCRIPTION (provided by applicant): The HIV epidemic has reached alarming rates in the United States especially within Black communities and is on par with those found in several African countries. HIV directly or indirectly related to men having sex with men is the single largest contributor to HIV infection among Blacks, accounting for nearly half of all cases among Black men. Despite large racial/ethnic disparities in HIV prevalence and risk, just one tested intervention specifically for Black men who have sex with men (MSM) has been published. Furthermore, Black men who have sex with both men and women (MSMW) have been particularly underserved and underrepresented in HIV research. Attention must be lent to address the behavioral, psychological, socio-cultural, and biomedical factors, as well as the racial/ethnic disparities that exist and contribute to HIV infection and transmission among Black MSM and MSMW. This 5-year R13 conference grant proposal entitled, Health, Hope and Healing (H3): A National Conference on HIV among Black MSM and MSMW, will assemble the best minds currently studying HIV/AIDS among Black MSM and MSMW. The H3 conference series, with an annual meeting over five years, aims to advance HIV biobehavioral research, mentor junior investigators, and foster community based participatory collaborations. The three measurable conference outcomes are to: 1) Produce an executive report detailing the long term research, training and policy agenda, highlighting recommendations designed to reduce HIV in Black MSM and MSMW;2) Produce an executive report detailing successful mentorship of junior investigators;and 3) Develop successful community based participatory research collaboratives, as measured by the submission of at least one joint proposal that operationalizes the research. The H3 Conference series will be among the first to discuss the needs of Black MSM and MSMW within a holistic and socio-cultural paradigm and disseminate research findings and healthcare and policy recommendations for eliminating HIV/AIDS within this population. PUBLIC HEALTH RELEVANCE: The HIV epidemic has reached alarming rates in the United States especially within Black communities. Researchers, clinicians, community members and congressman have all advocated for more attention to this epidemic. The proposed Health, Hope and Healing conference will gather the best minds currently studying HIV among Black MSM and MSMW. This conference aims to advance research, mentor junior investigators, foster community based participatory collaborations and disseminate findings and recommendations for the purpose of eliminating HIV/AIDS among Black MSM and MSMW.

Because of the slow pace of terrestrial ecosystem processes, including the slow generation time, growth rate, and decomposition rate of trees, the impact of changing climate and disturbance on forests plays out over hundreds of years. For this reason, centennial scale projections of terrestrial ecosystem models are used to anticipate the trajectory of forest response to environmental change. Modelers would like to have data on how forests have changed at regional scales and over hundreds of years to help assess such projections. A rich assemblage of relevant paleoecological data has been collected, but they have not been synthesized into a form that can be incorporated into broad-scale modeling efforts. Funding provided will support the establishment of a paleoecological observatory network (PALEON) to address this challenge. PALEON is an interdisciplinary team of paleoecologists, environmental statisticians, and ecosystem modelers with the goals of producing rigorous and robust reconstructions of forest change from the Atlantic to the Great Plains over the past 2,000 years, and then using these reconstructions to validate and improve the predictions of terrestrial ecosystem models.

PALEON has identified the integrated analysis of paleoecological data with statistical and mechanistic modeling as a key challenge for improving research capacity for anticipating the future of environmental change. For this reason, PALEON incorporates interdisciplinary training and community building into all aspects of the PALEON mission. In addition to focused working groups, PALEON works with relevant disciplinary communities to develop common approaches to data collection, analysis, and experimental protocols to ensure that long-term data can be seamlessly integrated into macroscale ecosystem analyses. Interdisciplinary training of post-doctoral fellows and graduate students, including a summer short course, will ensure that the next generation of researchers thinks naturally at the spatial and temporal scales relevant to understanding the broad scale impact of changing climate and land-use disturbance.

A. DEFINITION The Protein Identification and Localization (PIL) Core is a restructured core that has evolved from aspects of two previous cores, the Cell Biology and Cell Imaging Core which focused on cellular imaging and the Peptides and Proteomics Core which had a dual service of providing synthetic peptides and identifying proteins by mass spectrometry. While cell biological methods are now standard in most laboratories and peptide synthesis can be obtained both from University and commercial sources, what was unique about the two previous cores was their synergy in identifying and localizing proteins. These aspects are preserved and enhanced by the structure of the new PIL Core and build on the longstanding collaborative interactions between Drs. Williams, Andrews and Ernst. They also build on the fact that this type of research requires complex and expensive equipment not available in individual laboratories. The PIL Core is designed to provide information on the identity of proteins by mass spectrometry, either as individual proteins purified in an investigator's laboratory and usually separated as a band or spot by gel electrophoresis or globally as a mixture of proteins in a protein complex or organelle. Information is also be provided on post-translational modifications of proteins and on quantitative changes in protein content. Once a protein is identified, the Core can provide information on the localization of the protein in live and fixed cells and their changes over time and in response to physiological and pathophysiological perturbation. It is centered around microscopic imaging and quantitative analysis of digital information obtained primarily by laser scanning confocal microscopy (LSCM) and multiwavelength fluorescence imaging, although traditional electron microscopy of fixed and embedded specimens is also available for fine structure analysis. Modern biomedical research is characterized both by its interdisciplinary nature and by its dependence on increasingly sophisticated instrumentation and informatics. Although the goal is to look at a tissue, disease, or organism as a complex integrated system, we are still developing an understanding of the structure and function of the components which are often cell type dependent. With the mapping of the genome, attention has shifted to the more complex world of the proteome. Complete inventories of most mammalian protein complexes or organelles are yet to be completed and they often vary between cell types, during the cell cycle, and during various stages of development. Thus identification of proteins, their post-translational modifications, dynamic interactions and cellular localization, all in time and space and in a quantitative manner, is a daunting task that almost all laboratory scientists in the GI Peptide Center encounter in their research. These questions can be approached through the tools of the PIL Core, very often in concert with the Molecular Biology Core and national genomic resources such as the NCBI. The new PIL Core is built around expertise in the director's laboratories, and the availability of sophisticated instrumentation in three existing facilities, the Michigan Proteome Core, the Morphology and Imaging Laboratory (MIL) of the Department of Cell and Developmental Biology and the Michigan Diabetes Research and Training Center (MDRTC) which maintain facilities discussed in detail below for sophisticated mass spectrometry and high resolution microscopy. This arrangement is facilitated by the fact that Dr Andrews directs the Michigan Proteome Core, Dr Williams directs the Microscopy and Image Analysis Laboratory of the MDRTC and Dr Ernst has a long standing affiliation with the MIL. All three entities have established personnel and a recharge structure and are currently being use by GI Center members. While the Protein Identification and Localization Core will make use of the available highly sophisticated instrumentation, it also is built around the over 90 years of proteomic, cell biology and imaging expertise of Drs. Williams, Andrews and Ernst which has been primarily devoted to GI Tissues. Experienced Core staff, including Drs. Walker, Strahler and Mr. Nelson, will directly assist Center Investigators and carry out the procedures of the Core. All Core personnel will demonstrate and teach techniques to GI Center investigators, trainees and technicians. Finally another mission of the Core is to initiate, implement and disseminate new and innovative techniques in proteomics and cell imaging.

Research Objectives and Approaches: The objective of this research is to establish a versatile, designated light source in northern Alabama to support the area?s wide range of research activities encompassing optical frequency metrology, nanophotonics, plasmonics, micro-fabrication, and optical sensing. The approach is to create a service facility by acquiring an optical frequency synthesizer (OFS), along with compatible THz antennas, which will be accessible to the region?s academic, industrial and governmental research programs.
Intellectual Merit: The distinctive capabilities of the OFS will greatly benefit a number of ongoing research programs, including the development of microfluidic catalytic reactors, fiber-based plasmonic sensors and microresonator-based frequency combs. Moreover, a diverse array of new research activities, such as frequency-comb-based atmospheric clock distribution, probing of ozone-layer activities, study of intraband dynamics of semiconductor nanocrystals, and multi-wavelength-range trace gas sensing, will also be enabled by this novel laser system.
Broader Impacts: The new service facility is expected to become a catalyst that strengthens current partnerships between different sectors of the local research community and stimulates further collaborations across departments, research centers, institutions and disciplines. Meanwhile, the research activities enabled by the OFS will substantially enhance the quantity and quality of student research opportunities at both undergraduate and graduate levels. Furthermore, through developing stronger educational programs in science and engineering at UAH, this MRI acquisition will help build a knowledge dissemination ladder that bridges forefront research and K-12 education, and benefit a broad scope of the local community, including those from underrepresented groups.

DESCRIPTION (provided by applicant): Mu-opioid receptors are widely distributed in the central and peripheral nervous systems. The action of opioids in the mesolimbic dopamine pathway activates the reward pathway(s) that are a key point in the multistep process leading to abuse and addiction. Opioid receptors are expressed in multiple parts of the mesolimbic system and the action of opioids in one area of this complex system may contribute to, but will surely not dominate all aspects of the rewarding properties of opioids. For the past 20 years the action of opioids in the ventral midbrain dopamine system rested on the GABA interneurons of the VTA. Inhibition of those interneurons was proposed to increase the activity of dopamine neurons through a disinhibition. It is now clear that the action of opioids in the ventral midbrain is more complex. There are a number of GABA neurons that innervate dopamine neurons and many of those neurons express opioid receptors. Each of these afferent pathways can now be manipulated selectively with the use of optogenetic tools such that a comprehensive picture of opioid action can be obtained. This proposal will identify the opioid sensitive GABA inputs to dopamine neurons in naive and morphine treated animals. In order for disinhibition to be functionally relevant, there has to be a substantial amount of inhibition initially. The hypothesisis that the source of inhibition is dependent on the state of the animal. Each inhibitory pathway will be recruited differentially depending on the behavioral state of the animal. The proposed experiments will use brain slice experiments to demonstrate that the relative role of different GABA afferent pathways changes during withdrawal from chronic treatment of animals with morphine. Knowledge that different afferent pathways mediate reward and withdrawal will allow a directed approach to limit both reward and withdrawal.

DESCRIPTION (provided by applicant): This proposal requests support to construct and implement an electronic resource for information on the exocrine pancreas. This pancreas knowledge base will be called The Pancreapedia. It is designed to compile and curate knowledge on the exocrine pancreas, its component cell types and its diseases including pancreatitis and pancreatic cancer. It will also encourage new research and promote sharing of methods and research tools. It will contain six major divisions: Reviews, Pathways, Molecule Pages, Cell Structure, Research Tools, and People (Directory, community bulletin board). Content will be divided into Entries and Posts. Entries will be invited or submitted and managed by Editors and an Editorial Board that will carry out peer review. After acceptance, staff will format and mount the finished copy which will be accompanied by a version number, date and digital object identifier (DOI). Material will be hyperlinked to other relevant areas in the site and to sites outside such as PubMed and NCBI. Material can be updated when ever appropriate but when content is changed it will be given a new date and version number. Posts will be entered directly onto a structured web page by individuals who have registered as members and will include Directory Listings, Research Tools (mouse lines, viral vectors, antibodies) and Comments. Comments can be made on all entries and will be accompanied by the commenter's name and picture. The site is constructed using Drupal as the content management system with the aim of minimizing the work of putting up new content. It will be mounted on a server at the University of Michigan Library (MPublishing) which will be the Publisher and responsible for data security and integrity. All content will be open access with the copyright remaining with the author. After establishment of the resource funds to cover the cost of maintenance and putting up new content will come from The American Pancreatic Association and other organizations (Societies and Foundations) interested in pancreatic research.

Because of the slow pace of terrestrial ecosystem processes, including the slow generation time, growth rate, and decomposition rate of trees, the impact of changing climate and disturbance on forests plays out over hundreds of years. For this reason, terrestrial ecosystem models are used to anticipate the centennial scale projections of forest response to environmental change. Current terrestrial ecosystem model predictions vary widely and results have large statistical uncertainties. Furthermore, testing and calibration of these models relies on short term (sub-daily to decadal) data that fail to capture longer term trends and infrequent extreme events. The capacity of ecosystem models for scientific inference and long-term prediction would be greatly improved if uncertainties can be reduced through rigorous testing against observational data. PalEON is an interdisciplinary team of paleoecologists, statisticians, and modelers that have partnered to rigorously synthesize longer term paleoecological data and incorporate into ecosystem models to provide a deeper understanding of past dynamics and to use this knowledge to improve long-term forecasting capabilities.

Funds are provided to address four objectives and associated research questions: 1) Validation: How well do ecosystem models simulate decadal-to-centennial dynamics when confronted with past climate change, and what limits model accuracy? 2) Initialization: How sensitive are ecosystem models to initialization state and equilibrium assumptions? Do data-constrained simulations of centennial-scale dynamics improve 20thcentury simulations? 3) Inference: Was the terrestrial biosphere a carbon sink or source during the Little Ice Age and Medieval Climate Anomaly? and 4) Improvement: How can parameters and processes responsible for data-model divergences be improved? The data synthesis will include wide range of ecosystems, encompasses past climate variations that were large enough to affect tree growth rates, disturbance regimes, and forest demography, and leverages available paleodata. The synthesis will include 1) fossil pollen and Public Land Survey data to reconstruct forest composition, 2) sedimentary charcoal, stand-age and firescar indicators of past disturbance regimes, 3) tree-ring records of tree growth rates, and 4) multiple paleoclimatic proxies and paleoclimatic simulations. Bayesian hierarchical statistical models will be used to reconstruct key ecological variables and their associated uncertainty estimates. A standardized model intercomparison involving 13 ecosystem modeling groups will be used to evaluate the robustness of the modeling approach.

Three areas will be emphasized for PalEON's broader impacts. Community Building: The PalEON research community has doubled over the past 10 months, with more than 60 participants now. It is anticipated to nearly another doubling over the next five years, and the funds will allow the ongoing community-building via annual large meetings and task-oriented workshops. Interdisciplinary Training and Mentoring: A new generation of researchers will be trained to naturally conceptualize large spatial and temporal scales and to approach ecological forecasting as an integrative activity spanning data collection to model prediction. Eight postdocs and seven graduate students will be involved in proposed PalEON research, with multiple opportunities for cross-training. Additionally, the PalEON Summer Short Course provides an intensive cross-training experience for young scientists in all areas encompassed by PalEON. The 2012 course will be followed by courses in 2014 and 2016. Building Scientific Infrastructure: All PalEON datasets will be made publicly available upon publication, as will our new data-assimilation methods and model intercomparison protocols. Tools will be developed for optimal site selection (given the goal of reducing the integrated prediction uncertainty about past vegetation and climate over space and time) and will distribute a publicly available webtool version that will be linked directly to the Neotoma Paleoecology Database.

This project will provide new data on the paleoclimates, paleoenviroments and the biodiversity impacts of sea level rise on the southern edge of the Bering Land Bridge (BLB), and is intended to facilitate a better understanding of why woolly mammoths survived late into the mid-Holocene only in the environments of Arctic islands of this area. Furthermore, this research will attempt to establish the actual time of extinction of the Holocene mammoth population on St. Paul Island, Pribilof Islands, Alaska, and apply this information to test various proposed causal hypotheses for the extinction.

Cores from Cagaloq Lake, St. Paul, will be sampled for chironomids (aquatic invertebrates), pollen, coprophilous fungi spores, plant macrofossils, charcoal, ancient DNA and cryptotephras. Oxygen isotopes from the heads of chironomids will provide an independent climate record that can be supplemented by paleoecological analyses of the chironomid assemblages. Studies will also test chironomid species for isotope fractionation to disentangle climate signals from ground water effects. Terrestrial community changes will be reconstructed from pollen, spores and plant macrofossils; and charcoal frequency will be used to document fire events. Analysis of ancient DNA will provide data on cryptic plant and animal species that have not been detected by traditional methods of analysis, and can also be used to identify taxa to species.

Spores that grow on animal dung will be used as proxies for the mammoth population size and to document the time of mammoth extinction on the island. Ancient DNA will serve as an important cross-check by helping to resolve specific identifications of these spores as well as providing an independent estimate the time of extinction by the absence of mammoth DNA.

Digital elevations, bathymetric data, sea level curves, and Geographic Information System (GIS) technology will be used to reconstruct island size from the time of its isolation until today. A highly constrained chronology of the Cagaloq record will be achieved by using 14 C dates and tephras. All data can be compared temporally in order to test hypotheses for mammoth extinction.

This project will provide opportunities for two PhD students, a postdoctoral fellow, graduate student assistant and undergraduate student. Results of the study will be disseminated widely in peer-reviewed journals. An exhibit on the results of the project will be prepared by the EMS Museum at Penn State University with a web component. In addition, a special on-line, interactive exhibit will allow participants to reconstruct the island at various sizes, populate it with differing mammoth populations and define different climate and environmental factors to observe how each of these components affects mammoth extinction. The interactive exhibit will be available on the Neotoma database. Members of the group will make presentations on the project at the Alaska Quaternary Center in Fairbanks, which serves as a hub for promoting Alaska Quaternary research and outreach to the public.

This doctoral dissertation research project will investigate the role of climate change and land use in the emergence and decline of the complex agricultural society at Cahokia, the largest prehistoric Native American settlement north of the Rio Grande. Cahokia was located in southwestern Illinois in the Mississippi River valley near present-day St. Louis. Agriculture began to develop in eastern North America around 4,000 years ago, and by 1050 CE, Cahokia had emerged in the central Mississippi River valley with a population of approximately 10,000 inhabitants, monumental earthworks, and an expansive trade network. Despite its role as a cultural and political center, Cahokia was almost completely abandoned by 1350 CE for reasons that remain unclear. One hypothesis is that prolonged drought or severe flooding caused Cahokia's decline. Other prehistoric agricultural societies appear to have been highly vulnerable to periods of water scarcity that reduced crop yields and triggered social instability. Environmental degradation caused by agricultural intensification may have exacerbated the effects of climatic change and further contributed to Cahokia's decline. To assess the potential role of climate change and resource overexploitation in the emergence and decline of Cahokia, the doctoral student will collect lake sediment cores from four oxbow lakes in in the Mississippi River floodplain along a transect beginning at Cahokia and moving southward towards areas that were less populated during the late prehistoric period. From all sediment cores, the student will develop long-term records of agricultural expansion and contraction (using fossil pollen, charcoal, and carbon isotopes) and hydroclimatic variability (using sediment mineral content and particle-size) that span the last two millennia. These high-quality records of changing land use, land cover, fire regimes, and hydroclimatic conditions that permit tests of hypotheses concerning the role of environmental change in Cahokia's abandonment.

This project will contribute to a broader understanding of the resilience and vulnerability of agricultural societies to land-use intensification and climate variability. By developing an understanding of why past agricultural societies collapsed, this project will provide insights into how to manage contemporary agricultural systems in order to increase their resilience in the face of environmental perturbations. This project lies at the intersection of the earth sciences, ecology, geography, and anthropology, and the researchers will present their work at national conferences to both social and natural scientists. The results of this research will be shared with the public through presentations at the interpretive center at Cahokia. All data will be made publicly available through the Neotoma Paleoecology Database, an online repository for data from lake sediment cores. As a Doctoral Dissertation Research Improvement award, this project will provide support to enable a promising student to establish an independent research career.

The purpose of Flipping Calculus is to transform the teaching and learning of the material in Calculus. Flipping a course describes an instructional approach in which the delivery of a majority of the content is moved outside of class, via online videos, lecture notes, and readings, while homework, such as problem sets, labs, and applications, are shifted into the classroom. Instructors in Flipped Calculus are devoting class time to supporting small groups of students who are engaged in collaborative discussions and problem solving. Senior faculty are creating a library of videotaped lessons which correspond to key calculus concepts. In addition, they are developing and refining guided problem sets and assessments to evaluate the progress made by the students. Eventually, a website for flipping calculus will be created, which will host the videos and course materials, available for broad dissemination.

The project activities are based on research on how students learn and are designed to actively engage students in Calculus so they obtain a deeper understanding of the material, ultimately remembering and applying the skills and concepts they learn. This project involves at least six mathematics faculty members and is having a direct impact on more than 200 students during the first year of the project and upwards of 300 students in year two. The project leaders are extending the impact by presenting their findings and materials at regional and national conferences, sharing the videotapes, problem sets, labs and discussion questions on a website dedicated to flipping Calculus, as well as publishing articles in mathematics education journals.

This project establishes and operates the EarthCube Research Coordination Network (RCN)
Collaboration and Cyberinfrastructure for Paleogeosciences (C4P) to advance the role of cyberinfrastructure
(CI) in unraveling the large-scale, long-term evolution of the Earth-Life System through the
study of the geological record.Paleogeosciences research into our large-scale, long-term Earth-Life system is an ambitious
program that informs public debate on human stewardship of the earth and has - since its inception
- caught public imagination on issues such as species survival under environmental changes.
In the last decade, fresh discoveries, plus the application of new technologies of measurement,
data analysis, and modeling have revitalized the science. Bringing to bear recent advances
in computing and mathematics will further quicken the pace of discoveries, synthesis and publication
in the science. This RCN intends to stimulate and grow CI-focused collaborations and
partnerships among paleogeoscientists, paleobiologists, bioinformaticists, stratigraphers,
geochronologists, geographers, data scientists, and computer scientists that will help dissolve
existing intellectual barriers to dramatically improve the application of modern data management
approaches, data mining technologies, and computational methods to better analyze the heterogeneous
and sparse data of the Earth record in sediments, rocks, and ice within the paleogeosciences
and other domains and disciplines.

Activities envisioned include a series of coordinated workshops and webinars, web-enabled networking, outreach events such
as symposia and town-hall meetings, and cross-EarthCube coordination, and creation of an interoperable
C4P resource catalog that will be integrated with other EarthCube resource inventories. High-priority
themes in C4P are the management and representation of age and age uncertainty in data models
and computation to advance interoperability among and analysis of time-referenced data types,
the integration of physical samples into digital data infrastructure, and the need to properly
track and attribute provenance of data to ensure trust in the data and credit to authors.

How biological communities respond to environmental change is determined both by the tolerances of individual species and by interactions among species, such as competition, predation, and mutualism. However, little progress has been made in understanding how biotic interactions influence species distributions or in incorporating their effects into models of species assemblages at regional to continental scales. This project will develop and test new methods that consider species interactions by accounting for patterns of species co-occurrence to predict how species and biological communities respond to changes in climate. Observed changes in the distributions of plants and mammals in eastern North America during the last 21,000 years will be combined with independent simulations of past climates to examine how the strength and direction of species interactions vary across broad regions and in response to changes in climate. These data and analyses will also be used to test whether ecological models that consider species co-occurrences have a better predictive ability than existing approaches that model species individually and to identify possible limits to our ability to predict how biological communities may respond to future changes in climate. Results will help address the grand challenge of understanding how changes in climate alter natural systems and their associated ecosystem services.

Beyond providing important insights into how species interactions influence the response of biological communities to changes in climate and our ability to predict these responses, this project will also provide cross-disciplinary training of undergraduate and graduate students and early-career scientists in global change ecology, paleoecology, and statistical modeling. Minority and first-generation undergraduate students will be recruited through campus programs at Frostburg State University and the University of California-Merced. Public outreach to rural communities will be conducted through public lectures and engagements at open house events in an underserved region. In support of open, reproducible science, all publications, datasets, and computer code will be made publicly available.

This Partnerships for Innovation: Building Innovation Capacity project from Auburn University will be focused on building the basic foundations of a silicon-based high temperature integrated circuit technology. The final goal is to demonstrate a major polytype of silicon carbide (4H-SiC) metal-oxide semiconductor field-effect-transistor (that is, a 4H-SiC MOSFET)-based operational amplifier, operating at 250°C or higher. 4H-SiC is a wide-band gap semiconductor with a high critical breakdown field, high thermal conductivity, good bulk electron mobility, high chemical inertness, and mechanical hardness. These properties make 4H-SiC an extremely attractive material for electronics operating in harsh environments and at temperatures higher than 250°C, conditions under which conventional silicon-based electronics are very inefficient. In addition, the ability of SiC to oxidize to silicon dioxide, an insulator, naturally makes for MOSFETs. MOSFETs are voltage controlled electronic switches that are preferred over other devices as they offer a great deal of flexibility for circuit design. An efficient 4H-SiC MOSFET-based integrated circuit (IC) technology operating at high temperature will have a positive socio-economic impact in a variety of industrial and military applications. A limitation for employing 4H-SiC MOSFETs for this purpose is the low mobility of electrons in the conducting channel. This is primarily associated with a high density of traps at the oxide-SiC interface that results in carrier trapping and reduction of channel mobility. Recently, the Auburn group has demonstrated an advanced interface passivation process by incorporating phosphorus at and near the oxide-4H-SiC interface. This process results in a significantly reduced trap densities compared to the industry standard processes and accordingly results in at least a factor of two higher electron channel mobility compared to the state-of-the-art. The higher mobility is expected to result in significantly superior SiC operational-amplifiers. The intellectual merit of this proposal lies in the transfer of a fundamental materials science discovery to advanced applications taking future commercialization issues into consideration.

The broader impacts of this research are increasing U.S. technological competitiveness, increasing the business viability of small business partners, and developing students capable of contributing to the semiconductor industry. A high performance 4H-SiC IC technology has the potential to open a variety of new applications areas and markets. Some notable application areas are sensing and control circuits for geothermal, automotive and aeronautical sectors. Such technologies will have significant positive impact on U.S. competitiveness. The increased innovation capacity of the small business partners resulting from the success of this program would set the stage for building commercial prototypes to penetrate niche markets, increasing their business viability, and thus a future role in the nascent silicon carbide semiconductor industry. The success of such a technology would also create new possibilities for other end-user application-oriented businesses. An important aspect of the program is related to the education of graduate students. Students involved in this project will be exposed to the "food chain of semiconductor technology", ranging from basic semiconductor materials science to technology development and applications during their master's or doctoral research. Hands-on training related to a wide range of scientific and engineering problems will result in the development of young, highly trained scientists for the U.S. semiconductor industry. Additionally, the program will include workshops for high school students and teachers. Finally, existing close interactions with Tuskegee University, a local historically black university will be consolidated in this program.

Partners at the inception of the project are the lead institution: Auburn University; and two small technology-based businesses: CoolCAD Electronics LLC (College Park, MD), which performs analysis, design and prototyping for cryogenic, SiC and infrared (IR) electronics; and United Silicon Carbide Inc.(Monmouth Junction, NJ), which focuses on the design, fabrication, and commercialization of SiC technologies.

Morphology and Image Analysis Core (MIAC) The objectives of the core are to: 1. Provide consultation and training In microscopy and optical Image analysis for MDRC investigators Provide access to senior personnel highly skilled in microscopy and optical image analysis for MDRC Investigators 3. Provide access to state-of-the-art instrumentation for microscopy and optical image analysis for MDRC Investigators 4. Develop and/or implement new technologies for microscopy and optical Image analysis beneficial to MDRC Investigators. The MIAC remains focused on providing state-of-the-art quantitative morphological analysis of fixed and living cells for MDRC Investigators working on diabetes, its complications, and related endocrine and metabolic disorders. The Core provides service, consultation, collaboration and access to instrumentation for a variety of microscopic and analytic techniques. The Core focuses on light microscopic and confocal analysis of fixed and living cells using immunocyto-chemlstry, visualization of fluorescent proteins and use of fluorescent reporter probes such as Ca2+ and mitochondrial function. The Core's major instruments are three confocal microscopes and a widefield inverted fluorescence microscope, along with the Instrumentation and software to carry out quantitative and other. The capabilities of the MIAC have been substantially augmented during the current funding cycle.

DESCRIPTION (provided by applicant): The goal of the proposed research is to identify small molecule inhibitors of human metapneumovirus (HMPV). HMPV is a leading cause of lower respiratory infection in children worldwide, and recurrent infections occur throughout life. HMPV is associated with severe illness in older adults, with hospitalization rates similar to influenza, and persons with underlying conditions such as asthma, immune compromise, and chronic pulmonary disease. There are no licensed vaccines or drugs for HMPV and thus there is an unmet need for antivirals. We developed a fluorescent, cell-based high-throughput screening (HTS) assay for HMPV that captures inhibitors of all stages of the viral lifecycle except budding. We optimized and validated the assay, and performed a successful pilot HTS against ~3500 small compounds in the Vanderbilt Institute of Chemical Biology (VICB) HTS facility. We propose to conduct a HTS for HMPV inhibitors using a larger but focused compound set selected from the VICB library of >150,000 compounds. Bioinformatics tools will be used in conjunction with medicinal chemistry collaborators to prescreen and exclude known toxic compounds, and to enrich for structures related to the hits discovered in our pilot screen. We plan to screen 75,000 compounds. We will perform secondary assays to confirm hits; determine IC50, CC50, and selectivity index; and define mechanisms of action of inhibitors. The results of these experiments will identify new potential antivirals for HMPV and other respiratory viruses. Further, the findings will provide a foundation for SAR work to refine compounds and conduct preclinical studies. This project offers a unique opportunity for drug discovery against an important human pathogen. The compounds discovered will also provide powerful probes of viral biology.

Ecologists have long sought to understand how species reshuffle into novel ecological communities during periods of major environmental change, and the consequences of this reshuffling for the services that those ecological communities provide for human society. This project capitalizes on a major historical event - massive changes in plant communities roughly 17,000 years ago - to determine the relative roles that climate change, species interactions, and fire played in the creation of novel communities. Results from this research will inform contemporary debates about the potential for climate change to create 'no analog' communities that differ fundamentally in their structure, function, and the services they provide to society. They will also guide management practices intended to restore communities to prior states and improve our understanding of the consequences of widespread extinctions.

This project builds on prior evidence that plant communities changed dramatically during the last deglaciation because the extinction of large grazers released plants from herbivore suppression. New research will test the novel Megafaunal Release Hypothesis by assessing the relative importance of climate and species interactions on vegetation and fire dynamics. The investigators will collect and analyze a series of lake-sediment cores in the eastern US, and quantify relationships among independent climatic and ecological proxies within and across sites in order to test the Megafaunal Release Hypothesis. Eight new lake sites will be analyzed for indicators of past vegetation composition (fossil pollen), fire regime (sedimentary charcoal), megafaunal abundance (spores from fungi found in dung), and temperature (organic geochemistry). Broader impacts include the mentoring and training of a postdoctoral research scientist, a graduate student, and several undergraduates, and the expansion of collaborations among paleontologists, geographers, and ecologists to understand future environmental change. All data will be made publicly available through standard community data repositories.

? DESCRIPTION (provided by applicant): The role of dopamine in health and disease has been investigated for decades. Dopamine receptor agonists or antagonists are used for the treatment diseases such as schizophrenia, depression, attention deficit disorder and Parkinson's disease. Recent work also indicates that the altered regulation of dopamine release induced by many drugs of abuse play a critical role in early processes linked to early aspects of addiction. Thus there is a considerable understanding of the physiological role of dopamine in the central nervous system. Recent work indicates that dopamine neurons that project of various areas in the CNS are remarkably heterogeneous. The heterogeneity is the result of variable intrinsic properties due to differential expression of ion channels and transmitter receptors. It is also clear that afferent inputs to dopamine neurons vary within the ventral midbrain. Thus experiments from dopamine neurons with identified projections is a necessary step in obtaining an understanding of the mechanisms that regulate dopamine release in functionally different areas of the CNS. This proposal uses recordings from identified dopamine neurons in brain slices from mouse to (1) define the inhibitory synaptic potential (IPSP) that results from the dendritic release. (2) Identify the distribution of D2 receptors on the somato-dendritic membrane and relate that distribution to identified synapses distinguished by the presence of PSD95. (3) Determine how D2-receptor function and distribution are altered following treatment of animals with cocaine. The results of this study will form a connection between cocaine-induced plasticity at AMPA and D2 receptor dependent synaptic events. The characterization of these early events following treatment with cocaine will add considerably to the decades long search for an understanding of the role of dopamine in health and disease.

Paleontologists provide data about the past distribution and diversity of life. These data are useful both to geologists, because they can help determine the age of rocks, reconstruct past environments, and constrain models of the Earth system; and to biologists interested in the evolutionary history of organisms and the behavior of ecological systems during past global changes. Currently, data about fossils are dispersed across thousands of scientific publications, and dozens of small to large databases, only some of which are publicly available via the Internet. Even publicly available databases can be difficult to access because each stores different kinds of data with different conventions, requiring researchers to individually harmonize searches and their outputs. This project brings together six paleobiological databases so that they share a single set of Internet-based commands by which researchers and the public can easily access fossil records from all of Earth history. By coordinating with other emerging efforts in geological and biological data sharing, best practices, and protocols, we ensure that data will be freely available to all, enabling new scientific syntheses and discovery, more powerful educational opportunities, and general exploration of the history of life on Earth.

The paleobiological sciences sit at the nexus between geosciences and the biosciences, with close interdependencies in both domains. Within the geosciences, information about the past spatiotemporal distribution of organisms, species, and assemblages of species is essential to a wide array of allied disciplines: to sedimentologists and economic geologists studying facies relationships and employing biostratigraphic controls for correlating rock strata, to structural geologists and geophysicists seeking biogeographic constraints on reconstructions of former tectonic plate positions, to paleoclimatologists extracting paleoclimatic signals from paleoecological data, and to earth system modelers seeking to understand how biospheric dynamics have shaped, and continue to shape, the history of the Earth-Life system. Within the biosciences, the fossil record is essential for understanding how contemporary ecological systems are shaped by historical legacies of slow-acting processes, for testing climate-driven models of species distribution and diversity that are being used to project the impacts of 21st century climate change, for constraining phylogenetic models of species divergence and rates of evolution, and for understanding the fundamental drivers of biodiversity (i.e. species extinctions and originations). In an era of global change, when stewarding biodiversity is an urgent societal concern, conservation biologists, global change ecologists, and earth system scientists are all looking to the past to study the behavior of the Earth-Life system during rapid transitions. Paleobiological data are currently served by a wide array of databases that vary in structure, composition, temporal scales, types of data and metadata. To conduct ?global? or holistic analyses of the paleobiological record it is necessary to retrieve data from a variety of these databases - requiring queries of each database to retrieve the types of data needed. The purpose of this project is to make six different paleobiological databases interoperable so that they can be accessed via a common Application Programming Interface (API) to query the data from these and other databases. Towards that end, five key records of North American Pleistocene lakes will be uploaded and become available through this integrative project. This project also will increase the interoperability between these paleobiological resources and contemporary databases of species distributions and diversity, enabling continuous time-series analyses (e.g., of biodiversity) from the beginning of life on earth to today. Integration of the paleobiological databases with databases of the stratigraphic record (Macrostrat) will enhance the value of both types of data. New R packages will facilitate retrieval and analysis of data from all of the databases. Finally, this proposal establishes a Paleobiological Data Consortium, consisting of leaders of cyberinfrastructure resources in the paleobiosciences and allied disciplines, with the goal of sharing best practices and protocols among the geoinformatic and bioinformatic communities.

Project Summary / Abstract This program for pre- and post-doctoral training supports the training of specialists who are able to conduct preclinical research at levels ranging from the molecular to the cognitive/clinical on the biological mechanisms underlying the development, maintenance, and elimination of drug-seeking behavior. Twenty-three members of the graduate faculty of the Oregon Health & Science University (OHSU) serve as preceptors for postdoctoral research fellows and for Ph.D. students matriculating into basic science graduate programs in behavioral neuroscience, neuroscience, physiology and pharmacology, or biochemistry. Major research areas represent four levels. Some faculty members work primarily at the cellular/molecular level, using molecular biological, cell biological, and electron microscopic techniques. Other faculty work principally at the level of physiological, biochemical and pharmacological systems, using receptor binding, autoradiography, in vivo microdialysis and voltammetry, and electrophysiological techniques, and some work primarily in behavioral pharmacology and pharmacogenetics, using behavioral testing, intravenous drug self-administration, quantitative genetics and genetic mapping, as well as computer modelling techniques. Finally, some faculty work with human subjects, using cognitive testing and a variety of imaging techniques. Areas of extensive existing faculty collaboration include: studies of dopaminergic systems, ranging from molecular biology to behavior; extensive studies of genetic determinants of drug responses, at all levels from molecular to statistical gene mapping; and the study of learned and unlearned determinants of responses to drugs, particularly their rewarding effects and drug self- administration. Sensitivity, tolerance, and dependence/withdrawal phenomena for all major classes of drugs of abuse are under active investigation. Training includes firm curricular grounding in the basic sciences, specific pharmacological training in abused drugs, and extensive and continuous participation in research. Trainees also receive extensive instruction in the responsible conduct of research through interactive seminars that cover ethical considerations in all aspects of science. OHSU and this training program have put considerable efforts into recruiting and retaining trainees from diverse backgrounds. These efforts have resulted in a significantly more diverse pool of graduate students and postdoctoral fellows receiving training in the biological basis of drug-seeking behavior.

PROJECT SUMMARY The long-term goal of our research program is to understand mechanisms of human metapneumovirus (HMPV) immunity and pathogenesis and facilitate vaccine development. HMPV is a leading cause of severe lower respiratory infection (LRI) in children and adults worldwide. There is no licensed vaccine against HMPV. Recurrent infections with HMPV and other respiratory viruses such as respiratory syncytial virus (RSV) occur throughout life. Consequently, HMPV and RSV cause severe illness in older adults and persons with underlying conditions such as asthma, immune compromise, and chronic cardiopulmonary disease. Limited induction of CD8+ T cell (TCD8) memory by respiratory viruses may be a contributing factor to reinfection and presents an obstacle to the development of effective vaccines. RSV, influenza, and parainfluenza viruses have been shown to induce TCD8 with impaired cytolytic function and IFN? secretion, but the mechanism of this inhibition was unknown. Similar antigen unresponsiveness termed TCD8 exhaustion is associated with chronic viral infections and cancer. A key mediator of exhausted TCD8 is programmed cell death-1 (PD-1), a negative regulator of T cell activation implicated in maintaining peripheral tolerance and preventing autoimmunity. PD-1 and other inhibitory receptors including LAG-3 are therapeutic targets in chronic infections and cancer. A recognized adverse effect in these patients is respiratory inflammation, likely due to over-activated TCD8. We discovered that HMPV, influenza, and other respiratory viruses induce TCD8 impairment mediated initially by the PD-1 pathway. However, our preliminary data suggest that other inhibitory receptors, including LAG-3, are involved in maintaining lung TCD8 impairment in later stages of exhaustion. We hypothesize that respiratory viruses activate an orchestrated network of inhibitory pathways, which limit the acute TCD8 response and immunopathology, but also limit the induction of effective memory TCD8. These inhibitory pathways are likely natural immunoregulatory responses to limit lung inflammation, but which also limit effective host response and memory. Elucidating these mechanisms would increase understanding of poor immune memory against respiratory viruses and help guide vaccine development. In Specific Aim 1, we will test the hypothesis that PD-L1 has distinct functions on different cell types in the lung. In Aim 2, we will define the contribution of other inhibitory receptor pathways to late TCD8 impairment. In Specific Aim 3, we will test the hypothesis that lung TCD8 impairment programming can be bypassed using non-replicating virus-like particle (VLP) vaccines with local inhibitory receptor blockade. This work will elucidate mechanisms of TCD8 impairment and define the contribution of TCD8 immunoregulatory pathways in the memory response to vaccination and viral respiratory infections. The findings will guide HMPV vaccine development and identify biomarkers of effective TCD8 responses to vaccines. The results of these experiments will be applicable to other respiratory pathogens as well as relevant to respiratory complications of inhibitory receptor blockade therapy.

This Division of Earth Sciences Geoinformatics collaborative award supports continued development of Neotoma, a web-accessible centralized data clearinghouse for paleoecological data for which initial development support was provided by the EAR Geoinformatics Program (EAR- 0948652). This effort will enhance the current Neotoma platform by: 1) maximizing rates of data ingest through on-going data mobilization campaigns, recruitment and training of new data stewards, and development of facilitation software; 2) expanding the Neotoma data model to facilitate new proxy acquisition and secondary data including organic biomarkers, isotopic data from faunal specimens, and faunal taphonomic information; 3) developing new tools for data visualization and scientific exploration; 4) fostering engagement and growth of the Neotoma research community through support for a series of workshops intended to train data users, data stewards, and teachers, and by development of an on-line blog.

The Neotoma geoinformatics platform will promote integrative studies of the biotic impacts of climate change over the past five million years by allowing greater access to previously collected geoscience data sets of relevance and the tools to support analysis at marginal cost. The project will promote community engagement, support ease of data set ingestion, address PI team succession and promote educational module development. This award directly addresses OMB Memo M-13-13 ?Open Data Policy-Managing Information as an Asset.?

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This NSF Scholarships in Science, Technology, Engineering, and Mathematics (S-STEM) project will increase the number of students, who major in STEM disciplines at Albany State University and go on to successfully pursue STEM post-graduate education or research careers. During the project, 18 scholars will (a) develop and follow a four-year academic and professional plan that will include freshman learning community courses, workshops, service learning, and field trips or visitations under the guidance of a Student Success Coach, (b) participate in at least one undergraduate research experience, and (c) attend and present an undergraduate research project at an on-campus or regional conference. After graduation these talented scholars are expected to join the STEM workforce or go on to graduate school in STEM, enabling the US to compete and innovate in a global economy.

The theory of action for this project is that psychosocial factors such as STEM identity and self-efficacy are particularly critical to retaining STEM students. Thus, the project elements are designed to help students to develop these factors through a strong cohort anchored around a freshman learning community, targeted and intrusive advising, mentoring by faculty and peers, and participation in undergraduate research and internships. Evaluation will address the project's success in achieving its retention, course pass rate, and graduation objectives. Data analysis of student academic performance and participation in program activities will be used to study the effects of interventions like the learning community and intrusive advising on self-efficacy and science identity for students at a Historically Black University. The research results will be disseminated at the University System of GA Teaching and Learning Conference and the Understanding Interventions that Broaden Participation in STEM Careers conference. Manuscripts will be submitted to the Journal of Higher Education and the Journal of College Science Teaching.

This project is a workshop to advance interchange between ecologists working on general problems in modern systems and those focused on environments of the past (using buried pollen, plant and animal remnants, and other fossils). Study of the past is able to provide data on ecological dynamics that take place over long stretches of time, much longer than can be observed in current systems. It can shed light on processes that are important to current ecological change. However, it is often focused on the natural history of specific places and times rather than more general concepts. Meanwhile, ecologists working on general concepts in current systems are often unaware of what study of the past has to offer or are skeptical of whether the fossil data are adequate to answer their questions, given that experimental manipulation is not possible. By bringing both types of ecologists together the field of population and community ecology can be enriched. The outcome will contribute to the ability to respond to global change, and collaborations among scientists will be fostered.

Many of the anthropogenic impacts on populations and communities that are of high interest take decades or more to play out; sitting on the boundary between 'paleo' and 'neo' approaches, they require the methods and insights of both. The informatics revolution is also enabling the synthesis of multiple kinds of ecological data across spatial, temporal, and taxonomic scales, including species traits, past and present distributions, ancient and modern genetics, environmental covariates, and phylogenetic history. However, while there are clear opportunities for synergy between paleoecology and PCE, various structural challenges hinder integration between them. A workshop will facilitate capacity-building to address structural problems in the way that science is conducted that hinder the interaction of paleo- and neo- ecologists. Broader impacts will include networking and capacity building to strengthen the disciplinary ties between paleoecology and community ecology, which is critically important during a time of global change. This proposal facilitates critically timed career development and stabilization for diverse early career faculty.

Summary Acute respiratory illness (ARI) and acute gastroenteritis (AGE) are leading causes of disease in children in the U.S. and globally. These infections contribute a substantial burden of morbidity, mortality, and direct health care costs, in addition to the indirect costs associated with parental leave from work. Many cases of ARI are caused by viruses, including influenza, respiratory syncytial virus, and others. Similarly, most AGE in the U.S. is associated with viruses, including rotavirus and others. There are few or no effective antivirals for these, and therefore vaccination is the most promising intervention. Licensed vaccines are available for influenza and rotavirus; candidate vaccines for other pathogens are in development. Active, prospective surveillance is necessary to establish ?real-world? vaccine effectiveness (VE). Moreover, population-based surveillance can discover the burden of potentially vaccine- preventable diseases and guide policymakers and industry. The infrastructure provided by the New Vaccine Surveillance Network (NVSN) will facilitate these goals, as well as allowing the description of the clinical features, natural history, and population dynamics of these illnesses. The Children?s Hospital of Pittsburgh (CHP) offers an ideal environment to conduct the proposed population-based research, as CHP is the only major provider of pediatric inpatient and Emergency Department (ED) care in Allegheny County and the region. We propose three Specific Aims. Aim 1: To evaluate the effectiveness and impact(s) of current or upcoming vaccines and other immunoprophylaxis strategies, and inform pediatric vaccine-related policies. Using test-negative case-control methods, we will calculate the annual VE of influenza and rotavirus vaccines against medically attended ARI and AGE. Aim 2: To actively assess the burden of AGE and ARI (including illness with laboratory-confirmed viral etiologies) in the pediatric population. We will perform laboratory confirmation of viral etiologies of ARI and AGE among ill subjects enrolled in the inpatient or ED, and healthy controls enrolled at well-child visits. Aim 3: To establish the natural history of disease for pediatric infectious diseases, transmission dynamics, vaccine impacts for targeted and vulnerable populations, and socioeconomic and microbiological environments potentially relevant to public health interventions. We will capture extensive clinical and demographic data on enrolled subjects and healthy controls. Subjects will be tested for additional ARI- and AGE-associated viruses. The completion of this project will provide new data regarding the VE of licensed vaccines; define the population-based burden of potentially vaccine-preventable diseases; and establish the natural history and disease association of multiple human viruses. These results will guide the development of new vaccines and antivirals, inform public health policies, and enhance the health outcomes of children in the U.S. and globally.

Data heterogeneity and accessibility are major barriers to scientific progress. Many community curated data repositories (CCDRs) have emerged in the paleogeosciences in response to the needs of their scientific communities, but these CCDRs are not well integrated. This project seeks to transform geoscientific research by breaking down the barriers among the CCDRs that serve geoscientists. All participating resources are closely engaged with their respective disciplinary communities and each is mobilizing data from its communities; the key need is to facilitate data interchange among CCDRs. The work will align several major CCDRs using a system that develops new shared services that rely on common standards, and demonstrates the kinds of new scientific insights that become possible with an integrated geoscientific infrastructure.

This collaborative project will accomplish the following: 1) A survey of existing data structures and standards, their suitability for paleogeoscience CCDRs and alignment with requirements identified by the paleogeosciences research coordination network, and their degree of adoption. This will lead to recommendations for adoption by participating resources (EarthChem, Flyover Country, IODP, LacCore/CSDCO, LinkedEarth, Neotoma) and documentation written for geoscientific audiences. 2) Alignment of participating CCDRs to recommended standards and development of a common API that will allow data exchange among CCDRs and to third-party users. 3) Development of an Annotation Engine, which will provide a credentialed, crowd-sourced system for scientists to flag changes to datasets, to connect datasets post hoc, to add context to legacy data, and to provide link-back notification among CCDRs when linked dataset attributes change. Annotation Engine will be embedded into existing scientific CCDR-based workflows, minimizing disruption to users. 4) Development of GeoNoteBase to enable scientists to generate citeable, reproducible workflows that draw information from across data resources, with workflows made available through keyword searches, with full attribution. This is a pilot effort to begin some of the work Through two pilot scientific projects, THROUGHPUT will test and evaluate these new capabilities and demonstrate kinds of new scientific insights that can be gained through integration.

PROJECT SUMMARY The long-term goal of our research program is to understand mechanisms of human metapneumovirus (HMPV) immunity and pathogenesis and facilitate vaccine development. HMPV is a leading cause of severe lower respiratory infection (LRI) in children and adults worldwide. There is no licensed vaccine against HMPV. Recurrent infections with HMPV and other respiratory viruses such as respiratory syncytial virus (RSV) occur throughout life. Consequently, HMPV and RSV cause severe illness in older adults and persons with underlying conditions such as asthma, immune compromise, and chronic cardiopulmonary disease. Limited induction of CD8+ T cell (TCD8) memory by respiratory viruses may be a contributing factor to reinfection and presents an obstacle to the development of effective vaccines. RSV, influenza, and parainfluenza viruses have been shown to induce TCD8 with impaired cytolytic function and IFN? secretion, but the mechanism of this inhibition was unknown. Similar antigen unresponsiveness termed TCD8 exhaustion is associated with chronic viral infections and cancer. A key mediator of exhausted TCD8 is programmed cell death-1 (PD-1), a negative regulator of T cell activation implicated in maintaining peripheral tolerance and preventing autoimmunity. PD-1 and other inhibitory receptors including LAG-3 are therapeutic targets in chronic infections and cancer. A recognized adverse effect in these patients is respiratory inflammation, likely due to over-activated TCD8. We discovered that HMPV, influenza, and other respiratory viruses induce TCD8 impairment mediated initially by the PD-1 pathway. However, our preliminary data suggest that other inhibitory receptors, including LAG-3, are involved in maintaining lung TCD8 impairment in later stages of exhaustion. We hypothesize that respiratory viruses activate an orchestrated network of inhibitory pathways, which limit the acute TCD8 response and immunopathology, but also limit the induction of effective memory TCD8. These inhibitory pathways are likely natural immunoregulatory responses to limit lung inflammation, but which also limit effective host response and memory. Elucidating these mechanisms would increase understanding of poor immune memory against respiratory viruses and help guide vaccine development. In Specific Aim 1, we will test the hypothesis that PD-L1 has distinct functions on different cell types in the lung. In Aim 2, we will define the contribution of other inhibitory receptor pathways to late TCD8 impairment. In Specific Aim 3, we will test the hypothesis that lung TCD8 impairment programming can be bypassed using non-replicating virus-like particle (VLP) vaccines with local inhibitory receptor blockade. This work will elucidate mechanisms of TCD8 impairment and define the contribution of TCD8 immunoregulatory pathways in the memory response to vaccination and viral respiratory infections. The findings will guide HMPV vaccine development and identify biomarkers of effective TCD8 responses to vaccines. The results of these experiments will be applicable to other respiratory pathogens as well as relevant to respiratory complications of inhibitory receptor blockade therapy.

Brain functions are executed by intricately coordinated networks of neurons, whose modes of operation are highly sensitive to a constellation of neuromodulators. More specifically, neuromodulators such as dopamine, norepinephrine, serotonin, and acetylcholine exert dramatic control over global brain processes such as arousal, attention, emotion, or cognitive perception. Altered neuromodulator signaling has been linked to neurological and psychiatric disorders such as Parkinson's disease, schizophrenia, depression and addiction. Similarly, opioid neuropeptides play important roles in the modulation of cognition and behavior. While the anatomical structures that produce neuromodulatory signals are well known, little is known about the spatial and temporal evolution of these signals in the innervated brain regions. This is because current measurement techniques, such as microdialysis or cyclic voltammetry, lack the spatial or temporal resolution (and often the molecular specificity) to resolve respective signals. This technical challenge has been a long-standing barrier to our understanding of how neuromodulation alters neural circuit function in order to influence behavior. To address this challenge, this project will develop, validate, and disseminate novel genetically encoded fluorescent proteins for large-scale optical measurement of monoamine neuromodulator and opioid neuropeptide signaling in behaving animals, by bringing together a multi-disciplinary team of investigators with unique and complementary expertise. These sensor proteins have the potential to revolutionize neuroscience in a way similar to genetically encoded indicators for calcium, glutamate, and voltage, which are now in widespread use. Combined with calcium and voltage imaging, neuromodulator sensors will reveal how these systems impinge on cellular and circuit function. In particular, proposed sensors will enable minimally invasive, high-resolution, long-term, and direct measurement of neuromodulator and neuropeptide signaling at synaptic, cellular, and system levels. Sensors for neuromodulatory signaling have remained elusive for a long time. Our team recently developed a first generation of genetically encoded indicators for serotonin (5-HT), norepinephrine (NE), and dopamine (DA) that can report nano- to micromolar concentration changes with high spatial and temporal resolution. Building on this work, the following specific aims are proposed: 1) Optimize and diversify genetically encoded sensors for the monoamines using computational modeling, directed evolution and high-throughput screening; 2) Develop and optimize genetically encoded sensors for opiate neuropeptides using novel protein scaffolds; and 3) Systematically validate the novel neuromodulator and neuropeptide sensors in acute brain slices and behaving animals. Together, this work will provide the neuroscience community with a wide range of well-characterized multi-color indicators for probing the functional role of neuromodulators and neuropeptides in regulating neural circuit function and behavior in both health and disease.

The capability to bring computer science and technology as well as large and complex data sets to bear on interdisciplinary and transdisciplinary science is emerging. It is therefore critically important to establish and enable transnational frameworks so that data-driven scientific knowledge can transcend disciplines and geographical borders, ultimately increasing the scientific underpinnings of policy and action. International collaboration within global environmental change research fields holds the potential to establish international foundations for federated data integration and analysis systems with shared services, bring together best practices from the public and private sectors, foster open data and open science stewardship among the science communities including related areas such as publishing, and encourage data and cloud providers and others to adopt common standards and practices for the benefit of all.

This award supports U.S. researchers participating in a project competitively selected by a coalition of 29 funding agencies from 23 countries through the Belmont Forum Call for proposals on Science-driven e-Infrastructure Innovation (SEI) for the Enhancement of Transnational, Interdisciplinary and Transdisciplinary Data Use in Environmental Change. SEI is a multilateral initiative designed to support research projects that bring together environmental, social and economic scientists with data scientists, computational scientists, and e-infrastructure and cyber-infrastructure developers and providers to solve one or more of the methodological, technological and/or procedural challenges currently facing inter-disciplinary and transdisciplinary environmental change research that involves working with large, diverse and multi-source transnational data. The SEI call will intimately link research thinking and technological innovation toward accelerating the full-path of discovery-driven data use and open science and enable a broader scientific community to benefit from the identified new and potentially disruptive demonstrators or pilots toward solutions.

The historical record indicates that abrupt and unexpected change is the norm, not the exception and that these changes have direct consequences for many species and civilizations. Long‐term records from natural and historical archives have been essential to identifying these tipping elements, because past abrupt changes have occurred rapidly but infrequently, making them impossible to observe with instrumental records. The key limitations to addressing and communicating this major challenge in sustainability research is data access and incompatibility. Inaccessible or "dark" data, unstructured data, the lack of e-infrastructure to integrate multinational and multidisciplinary databases and datasets, are fundamental limit our understanding of abrupt change. This project, run by a diverse, international consortium of ecologists, climate scientists and informaticists will seek to build e-infrastructure that enables efficient cross-resource data access between trans-disciplinary and transnational data resources; and create an analysis package that allows users to detect, map and investigate abrupt change in Earth systems. The project will focus on determining the tipping elements in Earth's climate and ecosystems and to understand what drove rapid desertification in subtropical North Africa 6,000 years ago. In addition, the project will seek to model the FAIR data principles: 1) Findable: exposing dark data and developing tools for data discovery. 2) Accessible: transferring dark data to open-access platforms. 3) Interoperable: building cyberinfrastructure that enables cross-access. And 4) reusable: generating data synthesis products that identify essential metadata as a model for future data generators, while working with stakeholders to facilitate the development of community endorsed data standards.

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.

Opioid receptors are widely distributed in the central and peripheral nervous system and upon activation have manifold actions including; depression of respiration, activation of the reward pathway, disruption of normal gastrointestinal motility and analgesia. It is currently difficult to examine this widespread distribution in wild type animals. Antibodies against the opioid receptors are often very unreliable and the epitopes are most often intracellular such that detection in living tissue is not possible. The use of transgenic or knockin animals to identify receptors requires extensive characterization, can be limited by varied expression levels and has been limited to studies in mice. The goal of this proposal is to use a ligand directed approach to covalently label endogenous opioid receptors in living tissue. The guide compound, naltrexamine, is a non-selective opioid antagonist that has high affinity for all subtype opioid receptors. Naltrexamine will be coupled through a series of linkers, a reactive acyl imidazole and fluorescent ligand. Once bound to the receptor, the reactive acyl imidazole reacts with a lysine on the receptor and simultaneously releases the naltrexamine. Thus natrexamine is free to dissociate from the receptor, however the fluorescent ligand is left covalently bound to the receptor. Preliminary results indicate that this compound interacts very effectively with the mu-opioid receptor. Thus endogenous receptor are labeled and the labeling is not dependent on species. This compound and future refinements will be used areas of the CNS where opioid receptors are expressed in only a small population of neurons. This proposal will center on the VTA and Substantia Nigra in mouse and rat. Opioid receptor expressing neurons in living brain slices will be identified and whole cell recordings will be made. By targeting these cells a complete understanding of opioid action in this key area of the reward system will be possible. The results of this cell based study will address a long-standing question surrounding the mechanisms that underlie one aspect of the addictive properties of opioids.

The long term sustainability of federally funded research depends on the discovery, accessibility and reuse of data. However, data and research products are often stored in different locations. This makes it challenging to find and integrate related data. This project helps support the discovery of related but distributed research products for Earth science and natural history data. Researches will have a way to link data resources, add context, or provide additional information about data, software and publications. Researchers use this system to create annotations that link resources using unique identifiers. Over time, these links connect to create a network of data resources. This project will support the development of the underlying database, a user interface for access and discovery, and a number of documentation tools and workshops to help support the ongoing development and sustainability of the Throughput database.The broader impacts of this project include the engagement of early career researchers and the better sharing of data and other research products in the Earth sciences.

Improved discoverability of data, metadata and services is a need shared across the geosciences. A barrier that increases "time-to-science" in Earth Science research is the difficulty of integrating individual observations, concepts, data models, and statistical techniques across subfields. The Throughput Annotation Engine (TAE) offers a solution to the challenge of managing interdisciplinary workflows by providing multiple points of entry to access, annotate and interact with data, and to link code, data, publications, or other elements to one another. The TAE will support adoption as part of a system of systems, linking outside users to data repositories, and providing data stewards a degree of flexibility in deciding how to manage new information: whether to incorporate information in annotations into their data models, or to access and report annotations using the Throughput API. Throughput improves credit for data, software and documentation by providing a mechanism to track use and implementation across a range of publications and online resources, and will provide a new set of citation tools based on FORCE11 recommendations. Linked annotations in the TAE support the generation of metrics for research infrastructure beyond standard publication metrics. The Cookbook will allow individuals and communities to identify programmatic workflows, and link them to community data resources. The Cookbook will provide researchers with access to community best-practices, and, by leveraging ORCID user credentials, to individuals engaged in the development of documentation and workflows. Throughput offers a user-centered solution to deepening and densifying connections among the many nodes in the emerging linked data ecosystem of scientists, data, software services, journals, and funders.

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.

Our understanding of the past is being revolutionized by the study of ancient DNA recovered from fossils and sediments. Because many species do not preserve well in the fossil record, our knowledge is limited about the past locations of species. Ancient DNA can allow the detection of many species that lived in the past. Therefore, understanding how species migrate or otherwise survive past environmental changes can occur. Ancient DNA can also allow the detection of past losses of genetic diversity, due to species extinctions. However, as a data type, ancient DNA falls in a gap between databases designed to store genetic data and other kinds of databases designed to store fossil data. Because of this gap, researchers cannot easily assemble ancient DNA data to study biodiversity changes at large spatial and temporal scales. This project will support a workshop of leading researchers in ancient DNA, biodiversity databases, and paleontological databases, to build new systems for archiving and sharing ancient DNA data. The workshop will allow more open access to ancient DNA data by both researchers and educators. This access will enable more powerful integrative research approaches and authentic teaching experiences.

Specific workshop goals include: 1) review the current state of the art with respect to ancient DNA data; 2) review the current capabilities of existing cyberinfrastructures in genomics and paleoecology; 3) identify gaps and misalignments among existing resources; 4) establish priorities and initial standards for data and metadata reporting in community resources; and 5) envision the establishment of an informatics infrastructure for sedimentary ancient DNA. The people attending this workshop will maximize interdisciplinary exchange of ideas. The workshop will include early career researchers from groups traditionally underrepresented in science and technology. Ultimately, the workshop will produce guidance and resources for researchers working with ancient DNA.

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.

Project Summary Schizophrenia is among the most severe and burdensome medical conditions worldwide, yet the pathophysiological basis for symptoms experienced by patients remain unknown. This F30 application presents a research and training program that will support the applicant on a path towards becoming an NIH- funded independent investigator and physician-scientist focused on understanding the neurobiology of schizophrenia and related psychotic disorders. The activities in this application build on the candidate?s prior training and are set in a resource-rich environment that will facilitate the development of expertise in: 1) principles of clinical imaging in schizophrenia, 2) methods in cognitive neuroscience and functional magnetic resonance imaging (fMRI), 3) model-based analysis of auditory processing using fMRI, 4), fundamentals of clinical medicine and translational research, and 5) the responsible and ethical conduct of research. This research proposal aims to determine the relationships between auditory verbal hallucinations (AVH) experienced by individuals with schizophrenia and previous findings of: 1) impaired thalamocortical connectivity in mouse models of 22q11 deletion syndrome (22q11DS), the strongest known genetic risk factor for psychosis; 2) reduced resting-state functional connectivity between MGN and AC in patients with schizophrenia; and 3) impaired computational processes in auditory cortex and MGN. The overarching goal of the research to be carried out in this application is to use resting-state, task-based, and model-based multiband fMRI to identify impairments that correlate with the presence and severity of AVH in: 1) connectivity between medical geniculate nucleus (MGN) and auditory cortex (AC), and 2) auditory learning and percept generation in medial geniculate nucleus. I aim to study the following groups of individuals: unmedicated patients with SCZ and schizoaffective disorder (n=30) and matched healthy controls (varying degrees of subclinical perceptual disturbances; n=30). Using a thalamic localizer fMRI task, voxels comprising the medial geniculate nucleus will be identified in each subject. Subsequently, resting-state functional connectivity (RSFC) between MGN and AC will be determined using task-free fMRI. Using a model-based auditory discrimination fMRI task, neurocomputational underpinnings of auditory learning and percept generation will be measured. Neurobiological impairments detected via fMRI will be correlated with clinical measures of AVH severity. Such evidence would have important implications for our understanding of the brain bases of auditory verbal hallucinations and the pathogenesis of schizophrenia and would help generate new targets for treatment of this devastating illness.

SUMMARY (For Components A, B, and C). Acute gastroenteritis (AGE) and acute respiratory illness (ARI) are leading causes of childhood disease, accounting for a large proportion of hospitalizations, Emergency Department (ED) visits, and outpatient visits annually in the US. These illnesses are caused by diverse pathogens, including influenza, respiratory syncytial virus, human metapneumovirus, parainfluenza viruses, rhinovirus, enterovirus (EV), coronavirus including SARS-CoV-2, adenovirus, rotavirus, norovirus, astrovirus, and sapovirus. Moreover, some of these viruses are associated with other emerging childhood syndromes, including acute flaccid myelitis (AFM) associated with EV, and multisystem inflammatory syndrome in children (MIS-C) associated with SARS-CoV-2. Thus, viral AGE and ARI are of major public health importance, and result in serious long-term consequences for some children. There are few or no effective antivirals for these viruses and vaccination is the most promising intervention. Our goals are to conduct active, prospective population-based surveillance for AGE and ARI; to define the burden of vaccine-preventable diseases; describe the clinical features, natural history, and population dynamics; and establish vaccine effectiveness (VE) of licensed and impending vaccines and monitor VE over time. The New Vaccine Surveillance Network (NVSN) will facilitate these goals. We propose four Specific Aims: Aim 1: to conduct prospective active surveillance for AGE due to norovirus, rotavirus, and other enteric viruses among children seeking healthcare in ED, inpatient, and outpatient settings. (Component A, Mandatory Component 1; Optional Component B) Aim 2: to conduct prospective active surveillance for ARI due to respiratory viruses in these settings. (Component A, Mandatory Component 1; Optional Component B) Aim 3: to conduct prospective active surveillance for AFM syndrome in these settings. (Component A, Mandatory Component 2) Aim 4: to conduct prospective active surveillance for MIS-C. (Optional Component C) The Pittsburgh site has extensive experience with pediatric clinical research, including as a top enrolling NVSN site in the current NVSN cycle. UPMC Children?s Hospital of Pittsburgh (CHP) has a catchment area of >5.5 million people and admits >95% of hospitalized children in the surrounding county of >1.2 million. Thus, the environment is excellent for population-based research. The experienced investigative team includes experts from pediatric infectious diseases, critical care, rheumatology, and cardiology. The data and samples collected in this project will facilitate the capacity to calculate VE for multiple licensed and pending vaccines. The results of this project will inform best practices for diagnosis and treatment, guide vaccine recommendations, and determine public health interventions to prevent viral illness-related medical visits among children.