Patrick Walsh - US grants

At the present time it is impossible to predict how well patients with prostatic carcinoma will respond to hormonal therapy. However, if it were possible to make this prediction, those patients who are unlikely to achieve long term benefit could be selected for other forms of treatment at an earlier time and those patients most likely to respond could be spared the risk of unnecessary radiation or chemotherapy. It is the primary goal of this proposal to develop means to predict the hormonal responsiveness of prostatic cancer. Specifically, we propose to measure steroid receptor content (androgen, estrogen, progesterone), steroid content (testosterone, dihydrotestosterone), enzymatic profiles, and morphologic characteristics of the cancer cell nucleus in malignant prostatic tissue. These findings will be correlated with quantitative aspects of response to hormonal therapy to determine whether these measurements will be of predictive value. The response to hormonal therapy will be quantitated separately for both the primary prostatic neoplasm and metastatic deposits (bone and lung). It is our hope that these correlations will prove to be of predictive value so that guidelines can be established for the proper application of hormonal therapy in the management of men with prostatic cancer. As a secondary goal, these same parameters will be correlated with the response to shemotherapy in men with relapsing prostatic cancer and the disease-free interval in men following redical prostatectomy.

toxin metabolism; body temperature regulation; liver metabolism; liver cells;

toxin metabolism; body temperature regulation; liver metabolism; liver cells;

Dr. Patrick J. Walsh has been awarded a Postdoctoral Research Fellowship in Chemistry. Dr. Walsh's doctoral degree was from the University of California (Berkeley) under the supervision of Professor Robert G. Bergman. Dr. Walsh intends to continue research at the Massachusetts Institute of Technology under the sponsorship of Professor K. Barry Sharpless. Dr. Walsh's area of postdoctoral research and training will be the application of transition metal catalysis to synthetic organic chemistry. His long-term research interests are concerned with the asymmetric aziridination of olefins by transition metals. The Postdoctoral Research Fellowships in Chemistry Program is viewed as an important infrastructural program designed to broaden the knowledge and experience of new Ph.D.s and attract them into meaningful careers in contemporary chemical research and teaching.

This award is for repair and rehabilitation of the Glassell Building and its contiguous pen. This seawater facility is utilized by every scientific research program on the RSMAS campus that requires direct access to seawater. These programs include investigations in coral ecology, plankton ecology, pathology, physiology, animal behavior, bioacoustics, biochemistry and the culture of marine organisms. It also provides space and instrumentation for both graduate and undergraduate students being trained in one or more of these disciplines. This renovation is indispensable for continuing and expanding these and future programs at the School and is essential if they are to achieve and maintain state-of-the- art capabilities in experimental marine biology. Currently, users are handicapped by the difficult environment of a building that has served as the School's major seawater facility for 28 years. Within the last two years, the School has conducted a major renovation of the entire exterior of the Glassell Building, including various structural repairs, supervised and certified by structural engineers. These have assured that the building will be safe and secure for many years to come. This award will complete the renovation of the entire facility.

A major area of interest within the biological oceanographic community is the ability to assess the factors affecting population abundances of marine organisms and the potential role of climate change on marine species. For zooplankton species, it is hypothesized that nutritional regime can profoundly affect zooplankton abundance and survival. However, current techniques to assess nutritional and physiological condition in zooplankton to evaluate this hypothesis are rather insensitive and/or difficult to apply. Additionally, analysis of field data from biological studies usually lags behind the availability of physical measurements. This project will develop shipboard activity (rate) assays for enzymes of metabolism and growth in several species of marine zooplankton including two species (larvae of the cod, Gadus morhua and the copepod, Calanus finmarchicus) which are important components of an of the Georges Bank ecosystem, which is a region of interest in the zooplankton and fisheries community. The assays once developed will be used to test the vertical-stratification hypothesis for zooplankton survival in the Georges Bank region. The objectives will be accomplished by: (1) laboratory optimization of the enzyme assay conditions; (2) correlation of physiological parameters (e.g., respiration rates) with enzyme activities to validate molecular proxies; (3) estimation of physiological condition in field-caught specimens by use of these enzyme activity proxies.

The etiology of BPH remains obscure. Although aging and the presence of testes are permissive, these factors are not sufficient for the pathogenesis of clinical BPH. Autopsy studies have demonstrated that histologic BPH prevalence approaches 90% by age 80, but it is unclear at what point within this continuum and under what influence, microscopic evidence of hyperplasia becomes a clinically pathologic entity. There have been no large longitudinal cohort studies in humans that have used objective criteria to evaluate aging men with and without prostatic enlargement to gain insight into the pathogenesis, pathophysiology, and natural history of BPH. Elucidation of the pathogenesis of human BPH can identify potential new avenues of therapy and intervention. We propose to characterize dynamic aspects of BPH pathogenesis in a longitudinal study of prostate aging, to develop non-invasive techniques of assessing stromal and epithelial changes seen in BPH pathogenesis, and, expanding upon recent work demonstrating a genetic form of BPH, to conduct clinical genetic studies to identify genes involved in the pathogenesis of BPH. Longitudinal development of BPH will be evaluated in men participating in the Baltimore Longitudinal Study of Aging. Changes in prostate size, stromal epithelial ratio, serum PSA, and serum hormone profiles will be evaluated and their association with obstructive symptoms and response to therapy will be assessed. This study will build on preliminary data suggesting that PSA secretion by BPH tissue has unique characteristics, and will utilize a well established cohort of aging men. Recently developed MRI techniques capable of distinguishing between stromal and epithelial hyperplasia will be validated, optimized, and applied to the longitudinal aging cohort as well as to other men with clinical BPH in an attempt to prospectively determine predictors of clinical course. Epidemiological genetic studies will be undertaken in a national cohort of 4000 men with BPH and in a cohort of 10000 aging twins to confirm preliminary findings which identified a hereditary form of BPH. The potential role of hormonal factors in men with familial BPH will be evaluated, and linkage analysis will be initiated to identify genetic loci involved in familial BPH clustering. These studies aim to identify specific factors involved in the pathogenesis of human BPH and thereby identify potential new rational strategies of prevention and therapy.

9418360 Walsh This starter grant award to San Diego State University will support the research of Professor Patrick Walsh. The goal of the research is the development of new chiral metal complexes and their application as catalysts for organic synthesis. Specifically, ligands based on sulfonamide-substituted chiral diamines will be prepared which will form tetrahedral helical complexes when combined with Group IV metals. These strongly acidic complexes will be tested as catalyts for carbon-carbon bond formation activity in the Diels-Alder reaction, the Claisen rearrangement and aldol condensations. This study of asymmetric catalysis and molecular recognition will lead to advances in chiral Lewis acid catalysis. The new ligand complexes will be versatile and will be of general use in organic synthesis.

9507239 Walsh The broad outlines of the evolution of how animals excrete their waste nitrogen are reasonably well understood. Fully aquatic animals typically excrete ammonia as their main nitrogenous waste product because the water surrounding them will dilute the toxic ammonia. Terrestrial and semi- terrestrial animals, in order to conserve water, excrete nitrogenous waste products like as urea and uric acid that are less toxic and can be stored safely at higher concentrations. The production of these less toxic compounds is costly, however, because of their higher energy content. In spite of this expense, one fish, the gulf toadfish, has recently been shown to synthesize and excrete urea. This research is aimed at understanding the biochemical pathways the fish uses to make urea, their similarity to those of terrestrial organisms, and the genetic basis for this ability. Field studies will determine what advantage this ability provides in an aquatic environment. Preliminary results indicate that environmental stress induces the fish to make urea. A variety of biochemical, physiological, molecular and field techniques will be used both in the laboratory and in the fish's natural setting to identify synthetic pathways, control mechanisms, and the functional significance of urea excretion. This research should provide unique insight into the genetic mechanisms, physiological significance and evolution of an important physiological trait in vertebrate animals. Ultimately, it may lead to genetic advances that would enable culture of fish that make urea instead of ammonia that could simplify high-density aquaculture by reducing the need to control ammonia levels. If environmental stress is the trigger that initiates urea production, the abundance of the fish may make it a useful bioassay for biologically im portant changes in coastal water environments.

This proposal describes the establishment of a National Resource for Aplysia, whose overall goal is to provide consistently high quality cultured sea hares aplysia californica (and their cultured red algae food Gracilaria sp.) To NIH sponsored researchers. A. Californica is an important non- vertebrate (opisthobranch mollusc) model system for health related research, in the neurophysiology of behavior and learning. We will produce animals for research, and will conduct basic research aimed at exploring new model uses and at improving the resource. There are four sub-project/specific aims. 1. Production- We will increase our projected 1994 production by approximately 225% over the tenure of this grant and will make specimens from all life stages available at a price competitive with field-collected specimens. 2. Genetics- We will perform population genetic studies to characterize the amount of genetic variation present in our hatchery population in comparison to natural populations. The use of this genetic information will be to provide NIH-sponsored investigators with heterozygosity data for the hatchery population and with genetic markers for desirable/undesirable traits. 3. Developmental Neurophysiology-This project will investigate the regulation of neuronal excitability affecting reproductive development. The single cell voltage clamp technique will be applied to the study of the development of the capability of modulatory stimuli to act on ionic currents in applied to the study of the development of the capability of modulatory stimuli to act on ionic currents in neurosecretory bag cells. Development of this modulatory capability is an integral part of sexual maturation. This sub-project will further characterize the neurophysiology of immature A. Californica, potentially leading to a new model use, and will yield information useful to hatchery maturation and breeding strategies. 4. Animal Health Monitoring-We will establish a monitoring program based on screening of water quality and animal health parameters to assure rapid detection and complete documentation of any disease processes that might occur in any developmental stages of animal at the hatchery. Any disease syndromes and suspected pathogens observed will be investigated and appropriate control measures applied.

neurofibroma; disease /disorder model; model design /development; fish; biomedical facility; pathologic process; zebrafish; Aplysia; sharks; biological models; cellular immunity; genetics; histopathology; humoral immunity; saltwater environment; fresh water environment; aquatic biology; toxin metabolism; toxicology;

zebrafish; Aplysia; biomedical facility; animal colony; fish; water environment;

Malignant melanoma has one of the most rapidly increasing incidence rates in the U.S. Early detection and surgical excision can be curative, but once the tumor spreads beyond the skin it is one of the most deadly forms of cancer. There are currently no completely effective therapies for advanced (metastatic) disease and 10 year survival rates for these patients are very low. The objectives of this proposal are to conduct phase I clinical trials of a new gene therapy approach for the treatment of metastatic melanoma, to monitor for tumor responses attributable to treatment, and develop and characterize in vitro assays which may enable us to identify which patients are most likely to respond to this form of combined immuno-gene therapy. The therapeutic genes selected for use in the clinical trial are the genes for human granulocyte/macrophage colony stimulating factor (GM-CSF) and the gene for the superantigen staphylococcus enterotoxin B (SEB). In on-going preclinical trials conducted by one of the coinvestigators of this proposal, in a private veterinary oncology clinic, this combination of therapeutic genes has been shown to be more effective at inducing clinically significant tumor immunity than either gene used alone and more effective than other combinations of genes tested. The method used for gene transfection, polycationic lipid mediated gene transfection, has been tested extensively in the preclinical trials and found to be safe and effective. The phase I clinical trial is a dose escalation study designed to determine the safety and potential toxicities associated with the direct combination DNA injections of cutaneous melanoma metastases. Although the primary goal of the trial is to assess safety and toxicity, careful clinical analyses of treated and untreated tumors will allow the determination of whether the proposed treatments have an effect on local and distant metastases. In addition, in vitro immunologic assays which have proven to be indicative of development of protective tumor immunity in preclinical studies will be conducted concurrently with the clinical evaluations to determine if they have predictive value in identifying those patients which will benefit from this treatment approach. An improved understanding of how this treatment leads to the elimination of tumor tolerance and the development of immunity may enable us design more effective strategies for the use of these treatments in the clinical setting.

9724212 Reid This award to University of Miami in Florida provides funds for acquisition of a new scanning electronic microscope and related instrumentation. This field-emission environmental scanning electron microscope allows both high magnification sample analysis in high vacuum mode and analysis of wet or uncoated samples in low vacuum mode, significantly enhancing researchers' ability to study environmental samples without preparation artifacts. This award from the Major Research Instrumentation Program is jointly supported by both Division of Ocean Sciences in the Geosciences Directorate and the Division of Biological Infrastructure in the Biological Sciences Directorate. It is expected to provide major improvements over existing analytical capabilities for researchers in both earth and biological sciences, as well as other fields, and it is anticipated that it will serve as an important catalyst for integration of research and education in environmental sciences at University of Miami and other South Florida colleges. Substantial cost-share is offered by the University of Miami towards this project. ***

[unreadable] DESCRIPTION (provided by applicant): The broad, long-term objectives of this proposal are to develop new synthetic methods that enable the preparation of important intermediates for organic and medicinal chemistry. Our methods make possible, for the first time, the large-scale synthesis of enantioenriched diarylmethanols from aryl bromides. This method will be applied to the synthesis of enantiomerically enriched heterocycles bearing important pharmacophores, such as furans and pyridines. We have also developed several new routes to pyranones, furanosidic aldehydes, furans, and pyrroles. These medicinally important heterocycles can be prepared with high enantioselectivities using our methods. We have developed one-pot synthesis of a-hydroxy ketones, allylic alcohols, and substituted cyclopropanes from a versatile 1-alkenyl-1,1-bimetallic intermediate. These methods will be useful in complex molecule synthesis. The Specific Aims of this proposal are: I. Development of Direct Methods for Conversion of Aryl, Vinyl, and Alkyl Bromides to Enantioenriched Alcohols. II. Introduction of New Methods For Heterocycle Syntheses. III: Development of 1-Alkenyl-1,1-Bimetallic Reagents for Complex Molecule Synthesis. This project presents new methods to synthesize important chiral building blocks that were not easily accessible, but find utility in health-related applications, particularly in the synthesis of enantioenriched pharmaceutical agents. [unreadable] [unreadable] [unreadable]

This Faculty Early Career Development (CAREER) award in the Inorganic, Bioinorganic, and Organometallic Chemistry program supports research by Dr. Patrick J. Walsh, Chemistry Department, San Diego State University, on the mechanism of asymmetric induction by titanium catalysts. Several new chiral titanium bis(sulfonamido) complexes will be prepared and tested for their effectiveness in the catalytic asymmetric addition of alkyl groups to aldehydes. They are expected to model the intermediates in this reaction and so to provide guidance about important features of the active site and about design of more effective catalysts. In addition to involving undergraduate students in this research, Dr. Walsh will conduct an extensive outreach program to enrich chemistry education in the San Diego-northern Mexico region. Selective synthesis of compounds with a specific symmetry, that is, of either a right- or left-handed form, is an important chemical goal. Methods of this type are critical to the pharmaceutical industry and to many biological studies. In this research, a more detailed knowledge of the way in which selective synthesis occurs will be achieved. Undergraduates will be involved in this problem-directed research. In addition, outreach programs in science in collaboration with the Instituto Tecnologico de Tijuana for US and Mexican students will be undertaken.

superantigens; human therapy evaluation; metastasis; staphylococcal enterotoxin; gene therapy; interleukin 2; neoplasm /cancer immunotherapy; melanoma; vector vaccine; clinical trial phase I; combination cancer therapy; DNA; neoplasm /cancer vaccine; clinical research; human subject; injection /infusion; plasmids;

DESCRIPTION (provided by applicant) This proposal is to provide continued funding of an NIEHS Marine and Freshwater Biomedical Sciences Center at the University of Miami. Based at the Rosenstiel School of Marine and Atmospheric Sciences campus, the Center is it collaborative effort of 19 investigators from three University of Miami campuses and 6 external organizations. Two principal research themes form the basis for interdisciplinary Research Cores: "Marine and Freshwater Toxins and Human Health " and "Marine and Freshwater Animal Models Toxins and Human Health ". Both research themes are within the scope of NIEHS-sponsored research and contribute to the overall mission of the University of Miami Marine and Freshwater Biomedical Sciences Center: to evaluate the impact of the oceans and freshwater bodies on human health, by assessing and understanding risks, and by seeking remedies. The first research core includes: Toxin Biosynthesis and Probe Development, Metabolism, Molecular Pharmacology, Molecular Modeling, Electrophysiology Receptor Binding, Separation Techniques and Assay Methods, and Epidemiology and Public Health. The second research core includes several marine and freshwater (and human) models in various stages of development: Damselfish Neurofibromatosis, Cultured Human Schwann Cells, Aplysia Neurophysiology, Toadfish Hyperammonemia, Transgenic Fishes, Squirrelfish Zinc Metabolisin/Transport, Fish Immunology arid Sentinel Species. A vigorous Pilot Project Program is represented by four recently selected applications on: "Functional Analysis of Zinc Regulatory Genes in Transgenic Zebrafish"; "Red Tide Toxin Effects on Hearing: a Vertebrate Model"; "Molecule-Based Sensors for Carcinogenic Pollutants"; and "Microbial Recreational Water Indicators in the Subtropical Marine Environment". In support of existing individual and collaborative programs, 4 Facilities and Service Cores are proposed based on the investigators? evaluation of utilization and overall value to programs in the past five years: Toxin Probes, supplying brevetoxins, saxitoxin, okadaic acid, domoic acid, ciguatoxins and application-related toxin derivatives, as well as cultures of toxin organisms, and DNA-based materials from these organisms; In vitro and In vivo Fish Culture supplying facilities and expertise or the maintenance of, and experimentation with, fish and invertebrate cell cultures and live organisms; Analytical Chemistry and Electron Microscopy, which provides NMR, MS and analytical separation techniques assistance, and access to several EM methodologies; Neurophysiology, which provides two separate fully-equipped electrophysiology rigs, as well as additional tissue culture support capability. A well developed Community Outreach and Education Program will continue, including a poison-control hotline for seafood intoxication, an NIEHS-sponsored K-12 education program, and an NIEHS-sponsored postdoctoral training program. University of Miami will continue its commitment to the Center in the form of cost-share for salary support, instrumentation matching funds, and a faculty start-up package. Through the combined proposed NIEHS support and UM cost share, the investigators will continue to perform basic research on toxins and animal models, and to communicate the NIEHS message to the scientific and lay communities.

DESCRIPTION (Taken from the Applicant's Abstract) Miami-Dade County is home to more than 2.1 million people. Ethnic diversity is extensive, with a population that is 52% Hispanic, 34% African American, 13% White, and 1% American Indian/Asian/Other. As with any community of this complexity, there are significant environmental health issues of concern to the community and government. The Miami-Dade County Public School System is the fourth largest district in the country with more than 350,000 students, more than 93,000 of which are in grades 9-12. There is significant need within the public high school system to involve students with research scientists and members of the community in an interdisciplinary approach to learning about local environmental health science issues. The proposed AMBIENT Project (Atmospheric and Marine-Based Interdisciplinary ENvironmental Health Science Training) is a systemic approach to environmental health science education. Focused around the four environmental themes of air, water, soil and food, a health-science problem-based learning approach will be delivered by trained teachers to the ethnically diverse population of high school students in Miami-Dade County. The teachers will work together to enhance understanding of environmental and ethical issues through a hands-on summer workshop with research scientists from the University of Miami, Florida International University, and County Department of Health. Best practices from existing environmental curriculum materials will be assembled for use in the training. An important emphasis of the project will be to provide team teaching strategies for incorporating interdisciplinary activities into the large classes of more than 35 students at the high schools. The project is modeled after three highly successful environmental teacher training models, GLOBE, INSTAR, and the SECME Summer Institutes, and draws the best from each. Classroom activities and assessment tools will be incorporated by the Center for Educational Technologies at Wheeling, Jesuit University, into a problem-based learning web site similar to the NASA "Exploring the Environment" series. The Brown University Education Alliance will provide formative and summative assessment of the project. This project addresses the need defined by Priority 8.2 of Healthy People 2000: Educational and Community-Based Programs, which is to increase high school completion rates to 90 percent, especially with regard to Hispanic and Black American students.

Regulation of Ureotely in Batrachoidid Fishes
Patrick J. Walsh, M. Danielle McDonald
University of Miami


Until recently, it has been commonly accepted knowledge that fish and other aquatic species excrete ammonia as their main waste product following a meal. Ammonia is ultimately toxic, primarily to the brain, so fish and other aquatic species rid their bodies of ammonia as soon as it is produced, and its harmful effects are immediately diluted by the "infinite" volume of the surrounding water. However, when animals evolved to live on land, and did not have the diluting effects of a surrounding water environment, they needed to adopt alternate means of ridding their bodies of this toxin. So, terrestrial animals make (from ammonia) and excrete alternative waste products (urea and uric acid), which they can store in their bodies at higher concentrations without harmful effect until they are able to consume enough water with which to excrete them in their urine; the process of making and excreting urea is known as "ureotely". Surprisingly, in recent years, Walsh and colleagues have discovered that a common marine fish living in the bays and estuaries of the Southeastern US, the gulf toadfish (Opsanus beta), is an exception to this rule, opting to excrete urea instead of ammonia under certain stressful circumstances. Even more unusual, the toadfish excretes all of its urea in a single pulse lasting only a few hours across the gills. Thus the goals of this research project are to understand how this fish species is able to make and excrete urea, and to understand the ecological and evolutionary reasons for why it does so, and why it pulses its urea excretion. The ability to make urea requires a great deal of energy, which would otherwise be spent on predator avoidance, reproduction, etc., so it is reasonable to assume that the ability to make and excrete urea is somehow favored by natural selection, and contributes to the fish's ability to survive stressful conditions and propagate.

These goals will be approached with several different methods. In one portion of the study biochemical and molecular biology techniques will be used to understand how the fish shifts from making ammonia to making urea during stress, focusing on enzymes of urea production in the liver. In a second part of the study, physiological studies on how urea is excreted at the gill will be performed. In the third part of the study, field experiments will be used to test the hypotheses that urea production is important to the survival of the fish because the fish lives in an environment with high ammonia concentration, and/or that the pulses of urea excretion help it to be better at chemically camouflaging itself from predators. Lastly, studies will be conducted at the level of DNA to see how closely related species are within this family of fishes, and then compare members of the family that either do or do not adopt this mode of coping with stress. Through these "family tree" studies the PIs intend to determine where in the evolution of these species this trait has appeared or not, giving additional clues as to why the toadfish makes and excretes urea. The overall broader importance of the research is that it will help to understand how rapidly aquatic organisms are able to adapt to changing stressful environments on both an individual and generational time scale, and this work may eventually serve as a case study in the evolution of a complex physiological trait. This type of information becomes more and more valuable as our environment changes due to human influence. Furthermore, since ammonia is one of the most important waste byproducts in aquaculture that must be removed by expensive means to keep fish healthy, these studies may potentially suggest more efficient ways to culture fish and other aquatic species. This research will take place with significant international collaboration (with scientists from 5 other countries). Furthermore, since the University of Miami is a Minority Serving Institution, the PI will be able to recruit student researchers from minorities/underrepresented groups.

With support from the Chemistry Research Instrumentation and Facilities (CRIF) Program, the Department of Chemistry at the University of Pennsylvania will acquire a X-ray diffractometer with CCD detector for small molecule diffractometry. This equipment will enhance research in a number of areas including the following: a) synthesis and chemical and bioactivity properties of metallatricarbaboranyl and isoelectronic metallaheteroatomboranyl complexes; b) control of radical polymerization of olefins by metalloradical and atom transfer mechanisms; c) transition metal complexes of organosilicon and -germanium compounds: new structures and catalysts; d) conjugated porphyrin-based assemblies for biomimetic energy transduction and catalysis; and e)elucidation of catalyst structure and function.

The X-ray diffractometer allows accurate and precise measurements of the full three dimensional structure of a molecule, including bond distances and angles, and it provides accurate information about the spatial arrangement of the molecule relative to the neighboring molecules. These studies will have an impact in a number of areas, including the preparation of more efficient catalysts, materials chemistry and biotechnology.

0201438
Findley
This award to University of Miami's Rosenstiel School of Marine and Atmospheric Sciences provides instrumentation to improve the shared-use scientific instrumentation available to researchers using three research ships operated by University of Miami and the Harbor Branch Oceanographic Institution. These three vessels, R/Vs Seward Johnson, Seward Johnson II, and Walton Smith, are all operated as part of the University-National Oceanographic Laboratory System research fleet. The specific instrumentation supported by this award includes a phased array, 75 kHz acoustic Doppler current meter for installation on R/V Seward Johnson II, and a multi-corer device for collecting undisturbed seafloor samples from any of the three ships. These new capabilities will be of substantial advantage to marine scientists using the ships in their research during 2001 and future years.

DESCRIPTION (provided by applicant): Hepatic Encephalopathy (HE), and resultant elevated blood and tissue ammonia concentrations (i.e., hyperammonemia, HA), has profound central nervous system (CNS) effects, and can have environmental causes. In particular, liver damage due to exposure to toxicants such as carbon tetrachloride, toluene, DDT, heptachlor, etc., as well as chronic alcoholism and direct exposure to environmental ammonia, can elicit symptoms of HE/HA. However, there are such a wide variety of CNS effects produced in the disease in humans, and in rodent experimental models, that it is difficult to determine which disease biomarkers are the most critical indicators of disease progression. Furthermore, characteristics of the rodent model present several weaknesses in the study of HE/HA. Because of this gap in our knowledge, no practical and effective clinical intervention strategies are available to prevent or reverse biomarkers or symptoms of the disease. Recently, we have identified a vertebrate model, the gulf toadfish (Opsanus beta), which is both extremely tolerant of ammonia insult, and which, by virtue of its aquatic lifestyle, enables a line of experimentation not practical in mammalian models, namely rapid "ammonia washout" protocols. Therefore, we propose to test several hypotheses aimed at exploiting these and other characteristics of this new model to address the lack of biomarkers and intervention strategies for HE/HA. In particular, we will: (1) test the hypothesis that there are reversible vs. irreversible biomarkers of HE/HA, and that these can be readily identified and distinguished in an aquatic model like the toadfish; (2) test the hypotheses that extreme ammonia tolerance in the toadfish, relative to mammals, is due to an unusual aspect of its physiology, in particular, either to a more robust ammonia detoxification system in the brain, or to an inherent insensitivity of brain mitochondrial metabolism to ammonia insult. As a further test of this second hypothesis, we will also explore the possibility that the toadfish has higher levels of naturally occurring ammonia protectant compounds (e.g., carnitine, trimethylamine oxide, etc.) in its brain tissues than do mammals. In sum, these experiments will lead to information which is not readily obtainable from humans and existing mammalian models concerning the mechanisms of action of ammonia and cellular capacity for tolerance and recovery, and thus to a better understanding of the causes and mechanisms underlying HE/HA that could lead to therapeutic strategies.

The first objective of this research is to optimize and expand the scope of the asymmetric vinylation of aldehydes to furnish secondary allylic alcohols with high enantioselectivity. The second objective is to use the resulting chiral allylic alcohols in the synthesis of densely functionalized chiral building blocks. Using a single reaction vessel, tandem reactions resulting in the formation of multiple C-C bonds and stereocenters will be explored. Elaboration of the allylic alcohols to a variety of useful chiral building blocks including allylic amines, allylic cyclopropanes and allylic epoxides will be examined and the research will develop methods for the preparation of alpha- and beta- amino acids.

With this award, the Organic and Macromolecular Chemistry Program is supporting the research of Dr. Patrick J. Walsh of the Department of Chemistry at the University of Pennsylvania. Dr. Walsh will focus his work on the development of methodologies for the creation of chiral building blocks containing functional groups useful for directing reactivity. Specifically, chiral allylic alcohols will be targeted. The project could have important broader impact on organic synthesis, in particular in the pharmaceutical and agricultural industries. The work provides an excellent venue for the training of both undergraduate and graduate students and Professor Walsh has developed excellent international collaborations with Mexican scientists.

functional /structural genomics; technology /technique development; voltage /patch clamp

This project is focused on the study of stereoselective tandem reactions. There are three objectives in this project. The first one is to develop methods to generate (Z)-di- and (Z)-trisubstituted vinylzinc reagents. Tandem reactions will be studied to generate the (Z)-di- and (Z)-trisubstituted vinylzinc reagents with complete control of the double bond geometry. The second objective is to apply these new tandem C-C bond-forming reactions to the synthesis of key natural product subunits. The third objective is to study the tandem asymmetric addition/oxidation of furfurals to provide a variety of highly enantioenriched heterocycles.

With this award, the Organic and Macromolecular Chemistry Program is supporting the research of Professor Patrick J. Walsh of the Department of Chemistry at University of Pennsylvania. Professor Walsh and his students will be studying new methods to synthesize important organic intermediates and developing novel strategies for the synthesis of natural and unnatural products. Novel enantioenriched chiral building blocks will also be prepared in their efforts. Such building blocks will be useful for the synthesis of complex organic molecules of biomedical interest.

This project will continue work on the development of methods for the formation of C-C bonds using organozinc reagents, as the formation of C-C bonds is of fundamental importance in organic synthesis. New and efficient methods will be studied for the direct conversion of aldehydes into enals, which are among the most important starting materials for organocatalytic reactions. Methods for the stereospecific synthesis of enantioenriched enamines will also be explored. Finally, new classes of enantioenriched cyclopropanes will be prepared through novel halocyclopropanations.

With this award, the Organic and Macromolecular Chemistry Program is supporting the research of Professor Patrick J. Walsh of the Department of Chemistry at the University of Pennsylvania. Professor Walsh's research efforts involve the development of novel and practical methods for the formation of C-C bonds. Such chemistry will allow multiple reactions to be performed without isolation or purification of intermediates, thus reducing the environmental impact of chemical synthesis. Successful development of these methods will positively impact organic synthesis in the pharmaceutical and agricultural industries.

The GOALI project will integrate research groups at the University of Pennsylvania and members of the Merck, Rahway, Department of Process Research and the Catalytic Reactions Discovery and Development Laboratory. The focal point of this GOALI collaborative program is Merck?s High Throughput Experiments (HTE) facility, which will allow these two groups to collaborate on reaction discovery and optimization. Fundamental synthetic organic reactions of interest and importance not only to the pharmaceutical industry, but to the chemistry community as a whole will be explored to establish an inexpensive, permanent HTE facility at the University of Pennsylvania, which will in turn serve as a prototype that other academic institutions might adopt as a baseline tool for 21st century synthetic methods development.

With this award, the Organic and Macromolecular Program of the Division of Chemistry is supporting the groups of Professors Jeff Bode, Marisa Kozlowski, Gary Molander, and Patrick Walsh at the University of Pennsylvania, permitting them to advance their programs in synthetic methods development. Novel, more effective and environmentally sound methods for chemical synthesis are envisioned to arise from this program ? methods that can be implemented in the pharmaceutical, agricultural, and materials sciences sectors in the US and throughout the world.

With this award from the Chemistry Research Instrumentation and Facilities Multiuser Program (CRIF:MU), Professor Marsha I. Lester will acquire an X-ray diffractometer to be used by many users in the Department of Chemistry at the University of Pennsylvania as well as by some researchers from neighboring institutions. The main research projects that will benefit from this award are in areas of synthesis, catalysis, and materials. Examples involve NHC-carbene catalyzed reactions, peptide synthesis, design, characterization of organic cages that bind xenon with high affinity which are useful for biological imaging, identification of asymmetric organic compounds and perylenequinone natural products, polyborane compounds, asymmetric catalysis, and mechanistic investigations. The diffractometer will also be used in outreach activities involving K12 students and teachers as well as undergraduate and graduate students.

An X-ray diffractometer allows accurate and precise measurements of the full three dimensional structure of a molecule, including bond distances and angles, and provides accurate information about the spatial arrangement of a molecule relative to neighboring molecules. The studies described here will impact a number of areas, including chemistry, materials chemistry and biochemistry. This instrument will be an integral part of teaching as well as research.

In this International Collaboration in Chemistry (ICC) project funded by the Office of International Science and Engineering and the Chemical Catalysis Program of the Chemistry Division, Patrick J. Walsh and Eric J. Schelter, University of Pennsylvania, and Miquel A. Pericàs, Institute of Chemical Research of Catalonia (ICIQ), will link enantioselective lanthanide-based catalysts to solid supports, and enable their facile handling and recycling. Heterobimetallic lanthanide(BINOlate) complexes will be tethered to solid supports through diamine-containing polymers that will tightly bind to their lithium ions, resulting in solid state catalysts which can be used in asymmetric reactions with equal or better performance than their parent compounds. The supported catalysts will also be investigated in batch and flow reactors. Graduate and postdoctoral researchers will participate in exchanges between the Walsh and Schelter labs (U.S.) and the Pericàs group (Spain). These students will learn new techniques that are currently unavailable in their home institutions.

If successful, this work is expected to have far reaching implications in organic chemistry, polymer chemistry, and industrial and medicinal synthesis. This novel approach to design may lead to new solid catalysts that are able to perform key enantioselective transformations and then be easily separated from the reaction products (recovered) and reused in multiple applications. The development of flow systems is especially attractive as it provides evidence for the value-added nature of the assembled team in achieving practical, user-ready systems.

The MICINN will provide support funds for the Spanish investigators while the NSF will support the U.S. team's efforts.

In this project funded by the Chemical Synthesis Program of the Chemistry Division, Professor Patrick J. Walsh of the Department of Chemistry at the University of Pennsylvania will examine general methods for the chelation-controlled addition of organometallic reagents to alpha or beta silyloxy and alpha or beta halo aldehydes, ketones, and aldimimes. These substrates typically do not chelate. The central hypothesis of this project is that Cram-chelation can be achieved with substrates that strongly resist chelation by employing mild nucleophiles in combination with zinc Lewis acids. There are three specific objectives: 1. Developing chelation-controlled additions to chiral alpha and beta silyloxy aldehydes and ketones; 2. Assessing the ability of zinc reagents to chelate alpha or beta halo aldehydes and ketones; 3. Assessing the ability of zinc reagents to chelate alpha or beta halo imines and to promote Cram chelation additions.

The project could lead to general methods for highly diastereoselective additions to alpha or beta substituted aldehydes, ketones, and aldimines. These methods are highly useful for the preparation of a large number of substructures commonly seen in biologically important molecules, including syn-1,2- and anti-1,3-diols, syn-1,2-halo hydrins, cis-epoxides and allylic epoxides, and anti-1,2-amino alcohols. The successful results of this work could positively impact the pharmaceutical, agrochemical, and specialty chemical industries. In addition, this project will provide excellent training of students, including those from groups historically underrepresented in the sciences.

The vast majority of medications and biologically active natural products contain carbon- carbon bonds. The synthesis of these, and related small molecules, is therefore reliant on carbon-carbon bond-forming reactions. It follows that innovative approaches to efficient C-C bond formations will broaden the diversity of small molecules easily accessible and accelerate the discovery of drugs. The overall objective of this application is to develop novel and practical methods for sp3-C-H bond functionalizations/C-C couplings with very weakly acidic C-H's (pKa's 32 to over 40). Two mechanistic-based strategies are pursued to achieve this objective. The first involves activation of arenes toward mild deprotonation by coordination to transition metal catalysts, and encompasses enantioselective variants. Among the products of these reactions are tiarylmethanes and diarylmethylamines. The second approach is based on deprotonative cross-coupling procedures (DCCP's) wherein sp3-C-H's with pKa's as high as 35 are deprotonated under catalytic conditions and coupled with aryl halides. To perform this challenging class of reactions, new catalysts have been identified with unprecedented reactivity. By study of the mechanism of these catalysts, fundamentally new guiding principles have been revealed that are of interest to the greater chemistry community. This catalysts will be applied to DCCP's of unactivated diarylmethanes, allylbenzenes, N,N-dialkylbenzylamines, sulfoxides, sulfonamides, and sulfones to generate novel arylated products. The proposed method development, and mechanistic insight, will provide a collection of practical tools that enable new and efficient bond constructions that are easily applied to the synthesis of medications. The two approaches to C-C bond-formation in this application are innovative because they represent a clear departure from known chemistry.

In this project funded by the Chemical Catalysis Program of the Chemistry Division, Professor Patrick J. Walsh at the University of Pennsylvania is developing new reactions that form bonds between two carbon atoms and between a carbon atom and a sulfur atom. Bonds between these elements are found in pharmaceutical compounds, agricultural products, and polymers, and therefore, the proposed research has the potential to impact their corresponding scientific and technological areas. The overarching goal is to develop new catalysts and catalytic processes that are both synthetically useful and practical. Professor Walsh is also studying the reaction mechanisms (i.e., how the reactions work) and the factors that control the chemistry of the catalytic reactions, which will help to optimize the outcomes of the catalysis and to add to our fundamental knowledge base about chemistry.

Prof. Walsh is specifically addressing the following catalytic transformations: 1) The arylation of sulfenate anions and the enantioselective synthesis of diarylsulfoxides, which are found in a number of commercialized medications, 2) the development of methods to prepare sulfoximines, including in enantioenriched form, and 3) the introduction of a new class of organocatalysts, sulfenate anions, which have potential applications in a number of C-C bond forming reactions. The broader impacts of this research are in the following areas: 1) The new concepts and synthetic methods introduced herein are expected to have a positive impact on society as they may find application in the preparation of intermediates, medications, agricultural products, or materials in academics or industry, 2) Undergraduate students are gaining experience in chemical research, 3) The PI lectures for high school students about this NSF-supported research 4) The PI disseminates the findings of these investigations by publication and seminars at universities and conferences, and 5) The PI continues an international scientific collaboration outreach program with Mexican scientists.

NON-TECHNICAL DESCRIPTION: Nanoscopic thin films of small molecule amorphous organic materials are widely used in applications that range from protective coatings to organic photovoltaics and resist materials in nanoimprint lithography. These films are frequently manufactured through use of physical vapor deposition (PVD) onto a substrate held below the materials' glass transition temperature, Tg. Tg signifies the temperature where a system is unable to equilibrate on laboratory or computational time scales. Since glassy systems are out of equilibrium, the precise method of their fabrication, including substrate temperature, its properties, and rate of deposition can profoundly affect the materials properties and function in these applications. This project employs a combination of molecular synthesis, high-throughput characterization, and molecular simulation to design and characterize a library of synthetic glass-forming materials as a function of deposition variables. Addressing fundamental questions of the formation of highly stable glasses during PVD will have a transformative effect on the community's ability to engineer the properties of amorphous organic thin films and open the door to new applications of stable glasses for various industries. In addition to the project's impact on our fundamental understanding of stable glass formation and industrial applications, this project will impact the education of junior scientists from the undergraduate level through the PhD level. Undergraduate education is integrated into all aspects of the project. The starting material for the synthesis of glass formers is prepared as part of an undergraduate organic chemistry laboratory course. Advanced undergraduates and graduate students participate in the synthesis of the glass-formers as well as the characterization of PVD films using various experimental and computational techniques.

TECHNICAL DESCRIPTION: When held at a constant temperature a glass very slowly evolves towards a more stable, higher density state. This process, called physical aging, can take millions of years to reach equilibrium and only result in modest improvement in properties. Recent breakthrough studies have shown that PVD onto a substrate held just below Tg leads to a glass with properties that appear to be that of a glass that has aged hundreds or even thousands of years. It is hypothesized that this is a result of the enhanced mobility at the free surface of the film during deposition. Through PVD, each deposited molecule experiences this enhanced mobility upon condensation, allowing it to find a low energy state. As such, this process is referred to as surface mediated equilibration (SME). The remarkable kinetic stability of SME-generated glasses opens the door for their use in a number of new applications, but several fundamental challenges hinder their adoption. Most notably, a systematic understanding of the role of the chemical structure and intermolecular interactions, the interactions of the organic molecule with the substrate, and the effect of film thickness remain poorly understood. The synthesis capabilities previously developed by the PIs allows one to dial in particular structural motifs and intermolecular interactions. High-throughput characterization methods will enable rapid determination of a materials' kinetic stability as well as the relationship between stability and enhanced surface dynamics. Finally, molecular-level insights will be provided through coarse-grained simulations of the molecules synthesized and characterized experimentally. Specifically, the primary goals of this project are to i) determine the influence of chemical structure on surface mobility and SME glass stability; ii) determine the effect of film thickness on stability; and iii) determine the role of substrate interactions on altering materials' packing and ability to form a stable glass. Addressing these questions will have a transformative effect on the community's ability to engineer the properties of amorphous organic thin films and open the door to new applications of stable glasses.

With this award, the Chemical Catalysis Program of the NSF Division of Chemistry is supporting the research of Professor Patrick J. Walsh at the University of Pennsylvania. Professor Walsh is developing synthetic chemical methods to prepare molecular compounds using carbon monoxide (CO) and other more complex molecules as key reagents. The sequence of reactions employed prepares products that contain a "ketone" functional group, which contains a carbon bearing a double bond to an oxygen and single bonds to two other carbons. Ketones are found in numerous natural products and biologically active compounds, making them highly valuable intermediates and products. Ketones can be difficult to prepare and thus, they are the targets of this study. In addition to a streamlined method to synthesize ketones, the researchers are performing experiments that help them understand how the reactions occur. Other researchers can use these methods to design new reactions based on this fundamental understanding. Through this award, university students are involved in undergraduate research experiences in Professor Walsh's labs and will be co-authors on refereed publications. Upon graduation, these skilled researchers contribute to chemical industry, supporting the US economy. In addition, Professor Walsh gives annual lectures to high school organic chemistry students about his NSF supported research.

In this proposal, Professor Walsh is developing new catalysts, developing novel practical reactions, and delineating their reaction mechanisms. Much of the research involves the application of weakly acidic pro-nucleophiles in carbon-hydrogen (C-H) functionalization/carbonylation reactions under basic conditions. This approach avoids prefunctionalization and directing group strategies, enabling the efficient construction of C-C bonds. The objectives of the investigations include developing deprotonative carbonylation cross-coupling reactions (DCCC) and mapping the scope of weakly acidic substrates for DCCC reactions. The carbonylation of aryl halides is a powerful method to access esters, amides, aldehydes and ketones. Despite progress in the synthesis of ketones via such carbonylations, a significant gap in knowledge remains that prevents realization of its full potential. Current methods rely on carbonylation of aryl halides in the presence of preformed organometallic reagents. Drawbacks to this approach include the need to prepare or purchase these organometallic reagents, which can be toxic (organotins) or air/water sensitive (organoborons). The synthesis of these reagents adds costs and generates additional waste. The Walsh group is advancing practical approaches involving functionalization of pro-nucleophile C-H bonds in the carbonylation (1 atm CO) of aryl halides. This reaction is being performed based on mechanistic investigations and parameter screening. The scope and limitations of the method are being explored. In addition to the above-mentioned societal benefits, the methods developed herein are enabling other researchers to solve important synthetic problems. Furthermore, the new conceptual and synthetic methods may have a positive impact on the quality of life when used in the preparation of intermediates, medications, agricultural products, or materials in academics or industry.

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.