The recent controversial Women's Health Initiative Memory Study (WHIMS) implies that long-term treatment with estrogen and progestin significantly increases the risk of cognitive decline and dementia in menopausal women. These data sharply contrast with studies in women and rodents that have demonstrated a clear ability of estrogen to alleviate age- and hormone-related memory loss. The WHIMS finding that progestin is detrimental to memory is even more surprising because estrogen and progesterone treatment have been shown to be beneficial to memory in laboratory animals. The proposed studies were designed to clarify whether progesterone attenuates the beneficial effects of estrogen and to identify the neural mechanisms underlying the effects of estrogen and progesterone on memory consolidation. Because adverse side effects of hormone replacement such as increased risks of breast cancer and stroke make it impractical to conduct these studies in women, the proposed studies will utilize a female mouse model. The specific aims are as follows: 1) To pinpoint if estrogen and progesterone, alone or in combination, enhance memory consolidation and alter MAP kinase signaling in the hippocampus in young and aging females, 2) To determine if the mnemonic and hippocampal response to estrogen and progesterone is influenced by environmental factors, and 3) To compare the mnemonic and neural effects of estrogen and progesterone treatments that vary in duration (short- vs. long-term) and type (sustained vs. cyclic). To achieve these aims, ovariectomized female C57BL/6 mice (4, 17, and 24 months old) will be used in a series of nine experiments. Spatial memory will be tested in a spatial water maze task and non-spatial memory will be tested in an object recognition task. The goal of this research is to clarify the findings of WHIMS, a study which has greatly impacted the treatment of millions of menopausal women. WHIMS was a limited study, focusing on an older, highly educated population receiving treatment that was not cyclic and included a detrimental type of progestin. Establishing how factors such as age at treatment, degree of cognitive stimulation, type of progestin, and type of treatment (i.e., cyclic vs. sustained) influence the mnemonic and neural response to hormones will be critical to future use of hormone replacement. Our mouse model will enable a more controlled study of these factors than is possible with women. Elucidation of these issues may ultimately lead to the development of safer and more effective treatments to prevent cognitive decline and dementia in women [unreadable] [unreadable]


This application requests continuing Medical Scientist Training Program (MSTP) support for the Combined MD/PhD Training Program at the University of Minnesota. Originally funded in 1988, the Program remains committed to a single goal: to train physician-scientists equipped to push technology and clinical application to a level of investigative excellence that will have a direct impact on the diagnosis and treatment of human disease. The MSTP provides instruction leading to the MD and PhD degrees in approximately 8 years, although curriculum changes make completion in 7 years a feasible goal. The Program employs a sequential training sequence of medical school (2 years) > graduate school (3-4 years) > medical school (1.5 years). Students conduct a minimum of 3 laboratory rotations during their initial 18 months in the Program, and can choose from over 100 preceptors with research opportunities that span the gamut of disciplines in contemporary biomedical research, as well as biomedical engineering and epidemiology. Future research opportunities will be enhanced by a major expansion of laboratory facilities, including the construction of 4 new research buildings in the East Gateway Biomedical Discovery District at the University of Minnesota. Multiple requirements/opportunities facilitate the essential goal of integrating fundamental scientific training with the development of competent and compassionate physicians. A centerpiece of this integration is a course (Ambulatory Clinics for the Physician Scientist) in which students conduct clinical rotations with the physician-scientist of their choosing. Taken in the final year of graduate training, the course promotes thinking at a scientific/patient care boundary and refreshes students' clinical skills. Program leadership consists of a Director, Associate Director, and a Steering Committee made up of faculty from basic science and clinical departments. The Program recruits from a pool of gifted national applicants who can present a compelling case for a dual interest in science and medicine. The past 5 years have witnessed the complete elimination of any attrition problems, an enhanced curriculum that integrates scientific and clinical training, scientific accomplishments published in premiere journals (eg, Science and Nature), and residency choices that continue to promote academic career development. As of July 2009 there are 56 students in the Program (40% women, 14% under-represented minorities) from colleges and universities throughout the United States. These students are blessed with talent, highly productive (9 have individual F30 awards), and totally committed to a career in biomedical research. Our goal for the next 5 years is to continue our upward trajectory and witness the successful integration of our students into the fabric of biomedical research. RELEVANCE: This program supports the training of physician-scientists (i.e., persons pursuing an MD and a PhD). These individuals will be at the forefront of pioneering new ways to diagnose and treat human disease.


Hypothesis: A subpopulation of prolactinoma patients may have a syndrome of congential malformations consisting of pseudopapilledema and anomalous arterial supply of the pituitary by-passing hypothalamic dopamine inhibition. Exclusion of pituitary tissue from dopamine regulation may predispose a patient to prolactinoma formation. The current proposal proposes to investigate the incidence of pseudopapilledema in prolactinoma patients.


In the past 9 months, we have shown that rats can be operantly conditioned to increase as well as decrease the amplitude of EEG potential components recorded in somatic sensory face area cortex, and evoked by non-aversive shocks to the primary descending trigeminal tract. In 9 of 11 successful training procedures, we found that the latency of face-rub responses to noxious facial heat correlated significantly (p less than .05) with conditioned evoked potential amplitude, yielding correlation coefficients of between plus .5 and plus .9. We have also successfully conditioned potentials recorded in the caudal trigeminal nucleus, but there was no correlate in pain indices here.


This project is concerned with analyzing the differences in the Na-K pump and leak of cells of mammalian hibernators which enable them to retain normal Na and K concentrations at low temperature. Two types of cells are being investigated, erythrocytes and kidney cells, both grown in culture and freshly isolated, from both hibernating and non-hibernating species. With erythrocytes the aim is to determine quantitatively the specific effects of temperature on the kinetics and molecular mechanism of Na-K transport leading to its cessation at low temperature. In kidney cells some of the same analysis may also be possible, but in addition kidney cells should lend themselves to analysis of problems of regulation and turnover of pump proteins and of alterations of properties connected with transepithelial transport. Because of the latter possibility an effort is also being made to characterize the cultured cells in terms of their retention of differentiated epithelial transport activities. The general purpose of the project is twofold, both to understand the process of retention - or loss - of ionic regulation in the cold, which is a factor intimately involved with survival in the cold, and to use cells of hibernators to explore fundamental questions of transport.


Project Summary/Abstract: Evidence-based medicine (EBM) combines the best available practices with clinician experience and patient values. Prior to evidence-based medicine, quality of care was based on adherence to local standard practices given by healthcare providers. This standard was based on what historical practices, and was often anecdotal in nature. Improved means of statistical analysis, especially multivariable analysis, systematic reviews and meta-analysis has helped the advancement EBM. The adoption of EBM tenets has been slow, and acceptance of EBM over eminence-based orthopaedics has suffered. To this end, educating surgeons about the benefits and biases of high level of evidence studies, including randomized control trials (RCTs) and prospective observational studies, will help improve the clinical care of orthopaedic patients. Further, clinical interpretation in practice requires understanding of the Consolidated Standards of Reporting Trials (CONSORT) and Strengthening the Reporting of Observational studies in Epidemiology (STROBE) diagrams. The conflict between EBM and previously accepted standards is evident in several orthopaedic procedures in which EBM has questioned value: viscosupplementation, vertebroplasty, and knee meniscectomy for degenerative meniscal tears. Both RCTs and observational studies have important places in surgical research. A well-done cohort trial may provide greater insight to practical patient care than a RCT that is not applicable to many patients. In addition, observational studies can be particularly useful in addressing questions related to surgical procedures where RCTs cannot be performed or the number of patients necessary to achieve adequate statistical power is too numerous to make the trial practical. Improved education for surgeons reading and reviewing studies is critical to advancing the practice of orthopaedic surgery and the care of surgical patients. Improved understanding of the benefits and biases of studies at the highest levels of evidence, namely RCTs and prospective observational studies, will enable important and much needed advancements in the care of our patients. This symposium will address the strengths and weaknesses of these types of analyses in musculoskeletal care. The recent popularity of registry data for quality purposes will be explored as to how this fits within the hierarchy of evidence and physician/patient decision making. This is especially important given the AAOS emphasis and development of national registries for major orthopaedic procedures.


Dopamine has been implicated as the primary neurotransmitter associated with the psychomotor stimulant and reinforcing effects of cocaine. These findings have resulted in intensive efforts to characterize and elucidate the roles of the various dopamine receptor subtypes in the pharmacology and abuse liability of this drug of abuse. In this pursuit, the dopamine D3 receptor subtype has been recently targeted. However, definitive behavioral investigations have been hampered by the lack of highly selective D3 agonists and antagonists. We have prepared the known D3 antagonist NGB 2904 in multi-gram quantities for acute and chronic behavioral testing. This compound demonstrated in vivo D3 antagonist actions and attenuated cocaine self-administration under a progressive ratio schedule. NGB 2904 also blocked cocaine-induced reinstatement of drug seeking behavior, an animal model of relapse. We then designed and synthesized novel series of compounds, based on NGB 2904. All the compounds included either a 2,3-dichloro- or 2-methoxy-substituted- phenylpiperazine, a four carbon linking chain with varying saturation (butyl, hydroxybutyl, and trans butenyl) and a terminal aryl amide. Evaluation for in vitro binding in HEK 293 cells transfected with human D2, D3, or D4 receptor cDNAs resulted in D3 binding affinities ranging from Ki=0.3-500 nM. The most potent analogs in this series, demonstrated D3/D2 selectivity of >200 and a D3/D4 selectivity of >1000. Functional evaluation in vitro using a mitogenesis assay in D3 or D2 receptor transfected CHO cells demonstrated that these compounds were either potent antagonists or partial agonists at D3 receptors and were selective over D2 receptors, in this function. However, a functional comparison of a series of butenyl and saturated butyl analogues showed that these compounds generally exhibited higher intrinsic activity in the adenylate cyclase assay than in the mitogenesis assay, suggesting the potential of functional selectivity. Furthermore, structure-activity relationships (SAR) were deduced based on function, which was both instructive and provides additional functional data to be compared to in vivo activities for the identification of underlying mechanisms, at the G-protein level. In binding studies, SAR demonstrated that the trans-butenyl linker provided additional D3 selectivity as compared to the saturated linking chain. Moreover, addition of a hydroxy (OH) group in the 2- or 3-position of the butyl linker also gave several highly selective and potent D3 antagonists or partial agonists. Further, replacement of the sterically bulky aryl ring system with various heteroaryl groups served to retain high affinity and selectivity for D3, while decreasing lipophilicity. To this end we have recently discovered very selective D3 antagonists and partial agonists with D3/D2-selectivites reaching 400-fold. In addition, several of these analogues have been further screened for binding in 60 additional receptor and ion channel assays and did not show significant binding affinities at any of these other targets, highlighting that these agents are some of the most potent and selective D3-antagonists and partial agonists reported to date. Further, the (+)- and (-)-enantiomers of one of these 3-OH analogues, PG648, were synthesized and demonstrated enantioselectivity at D3 (>15-fold), but not significantly (<2-fold) at D2 receptors. This was the first demonstration of enantioselectivity of a D3 antagonist and further chimera studies, with these enantiomers, identified a transmembrane region that appears to differ between D3 and D2. Further characterization of these enantiomers and the synthesis of others are currently underway. The latter goal of reducing lipophilicity of the most potent agents was to improve physico-chemical properties that would provide a more favorable pharmacokinetic/bioavailability profile than the currently existing D3 agents. Ten of these analogues are currently being evaluated for pharmacokinetics, blood brain barrier penetration, and for potential metabolic pathways for degradation in vivo, in rats. These compounds are all being tested in a D3-agonist induced yawning model, in rats, to compare their pharmacological and bioavailability profiles in vivo. In addition, several of the most potent and selective compounds of this series have been synthesized in multi-gram quantities and are currently being evaluated in numerous animal models of cocaine and methamphetamine abuse, in both rodents and primates. Chronic studies in these and additional rodent and monkey models of drug abuse and impulsivity are underway, with PG01037 and several other butenyl and hydroxy-butyl linked analogues. In addition, another of these highly selective 3-OH analogues, GCC 3-09 was recently tritiated. 3HGCC 3-09 is currently being developed as a potential radioligand for both rat brain tissue and cell-based binding assays. Our newest series of analogues replaces the 3-OH group in the butyl linking chain with a F-group, and several of these compounds show favorable pharmacological profiles in vivo, with D3/D2-selectivites >200-fold. In addition, we have embarked on an SAR study of novel D2 antagonists, based on the parent ligand L741,626. Structural modification of this molecule has revealed important structural features that impart high affinity for D2 receptors, although improving D2/D3 selectivity has thus far remained elusive. Nevertheless, exceptionally high D2/D3 receptor affinities with some structural motifs have led to the hypothesis that our highly D3-selective ligands maybe bitopic and thus accessing both orthosteric and allosteric binding sites on the D3 receptor. Current synthesis and pharmacological assay development to test this hypothesis is underway.


Fourteen Chinese and sixteen Caucasian normal volunteers were administered a single dose of desipramine (DMI). Plasma and urine samples were analyzed for DMI and an active metabolite, hydroxydesipramine. The total clearance of DMI was lower in Chinese was lower than in Caucasians but the clearance to hydroxydesipramine did not differ between the groups. The renal clearance of hydroxydesipramine was lower in older subjects.


We hypothesize that hypothalamic inflammation associated with high fat feeding arises as a result of Toll-like receptor (TLR) signaling in a specialized cell type lining the inferolateral walls of the third ventricle, the alpha tanycyte. We propose that the activation of NF-?B in these cells releases inflammatory mediators into feeding-related centers in the mediobasal hypothalamus, increasing feeding and/or altering the sensitivity of arcuate/ventromedial nucleus neurons to leptin and/or insulin. To test this hypothesis, RNA-Seq analysis will be used to identify the complete gene expression profile of alpha tanycytes in response to a HFD, with particular emphasis given to the identification of genes and pathways involved in inflammation. Based on this analysis, the effect of a HFD on food intake, energy expenditure, glucose homeostasis, leptin and insulin sensitivity, and other parameters of hypothalamic inflammation will be studied in transgenic animals with conditional KO in alpha tanycytes of selected genes identified in the RNA-Seq analysis. We propose that these studies will demonstrate a critical role of tanycytes in the pathophysiology of hypothalamic inflammation associated with high fat feeding, uncovering new knowledge in understanding the pathogenesis of obesity and a potentially new pharmacologic target for the treatment of obesity.


The central goals of this proposal concern the relationship between protein structure and protein function. The powerful techniques of recombinant DNA technology will be applied by four investigators to study distinct groups of biologically important macromolecules. Most of these molecules are involved in cell-cell communication and in most instances, DNA probes are available to allow a more global approach to a full appreciation of the synthesis, expression and activity of a particular protein. The Program Project mechanism is requested as the technologies required to carry out each individual project require not only the intense intellectual interaction presently taking place, but, in addition, the pooling of resources and equipment. All the studies largely involve protocols in the mouse and rat, although the results of these projects will have clear applications to man.


The Case CCC Imaging Research Core Facility provides researchers with state-of-the art facilities and the services for in vivo imaging. The facility provides cancer researchers its extensive imaging research infrastructure and expertise of -26 imaging faculty in Radiology, Chemistry, Physics, and Biomedical Engineering to enhance and expand ongoing research efforts. The advent of dedicated small animal imaging systems allows the integration of in vivo physiologic measurements with microscopic measurements of structural and cellular activities. The collaborative, multidisciplinary approach provides a wealth of new information in elucidating the complex relationship between structure, genetics, replication, and function in genetically manipulated animal models of disease that have been used widely in cancer research with grounding in cellular/molecular-level understanding. The Imaging Research Core Facility is the centerpiece of the Case Center for Imaging Research, which encompasses all aspects of the much broader imaging research program at Case/UH. This includes molecular imaging, small animal imaging, and clinical imaging thrusts. The Core provides services to Case CCC members from 7 of 8 research programs, with members from the Imaging Program and Cancer Cell Signaling utilizing the Core's services the most. The Core has been actively involved in studies of the glioma microenvironment where novel cryo-imaging techniques demonstrated migration and dispersal pathways in vivid three-dimensional detail. In addition, fluorescence imaging systems in the Core were used to discover that while activation of EphA2 with its ligand ephrin-AI inhibits chemotactic migration of glioma and prostate cancer cells, EphA2 over-expression promotes migration in a ligand-independent manner.


The Baltimore Cancer Research Program (BCRP) has developed cancer guidance displays (treatment protocols) on the computerized Problem Oriented Medical Information System (PROMIS) in Burlington, Vermont at the University of Vermont School of Medicine, with the aid of a communication linkage between BCRP and PROMIS. The computerized displays for the logic and action of treating cancer problems are defined by the creation and sequencing of the computer display pages or frames. These displays provide an automatic connective device between the four phases of medical action of the Problem Oriented Medical Record. The problem-defining sequence determines the specific plan and it is that plan which is in part a protocol or series of protocols for treating a specific cancer problem. It is through the use of these explicit information and directional displays that the health care provider can couple learning with doing while gathering information and treating patients' cancer problems. The entire systems package has been shipped to BCRP and testing and debugging has begun in conjunction with the Interagency Agreement with the National Center for Health Services Research (Y01 CM 80109). This project ends on May 31, 1981.


Mammalian stem and progenitor cells activate the expression of specific regulatory genes to establish and stabilize different cell fates, but the mechanisms and general principles controlling this process are not well understood. Cells utilize fate-specifying regulatory genes to establish and maintain distinct fate identities during development, but it is not clear how they activate and maintain the expression of these genes to establish fate identity. Here, I propose to study this question in the context of two systems: T-cell fate commitment, which is driven by the activation of the T-cell specific transcription factor Bcl11b (Aims 1 and 2); and macrophage development, which we recently found is driven by the cell-cycle length dependent accumulation of the myeloid transcription factor PU.1 (Kueh et al., 2013)1 (Aim 3). Using these two systems, I will test a number of widely debated ideas in the field of developmental gene regulation. First, I will test the idea that developmental signals directly activate the expression of regulatory genes to instruct cell fate (Aim 1). Next, I will tet two proposed classes of mechanisms for stabilizing regulatory gene expression and fate identity: cis-acting mechanisms involving stable and heritable epigenetic modifications at regulatory gene loci (Aim 2); and trans- acting mechanisms involving self-reinforcing positive feedback loops on regulatory gene expression (Aim 3). My main approach will be to use timelapse live-cell imaging to track the expression dynamics of Bcl11b during T-cell development, and PU.1 during macrophage development. As cell differentiation is a dynamic and intrinsically heterogeneous process, single-cell tracking by timelapse imaging will reveal insights that are difficult to obtain with conventional discrete time-point population measurements. To gain mechanistic insights, I will perturb the mechanisms under investigation, and measure their resultant effects using timelapse imaging. These perturbations will involve over-expression or knockdown of genes; for studies of cis-epigenetic mechanisms, I will also develop a CRISPr-based system for perturbing chromatin marks at specific sites in the genome. To better understand this experimental data, and generate predictions for future experiments, I will then use mathematical modeling to analyze the behavior and dynamics of the different regulatory mechanisms studied. Finally, I will complement these approaches with genome-wide measurements of gene expression states in developing cells using high throughput sequencing, which will provide a more global picture of developmental changes, and potentially yield new directions for future work. Through these studies, I hope to uncover fundamental insights into how mammalian cells establish and maintain their distinct fate identities. These insights will potentially help us develop new therapies for leukemia and other cancers, and help us better manipulate stem cells for regenerative medicine.


: This application proposes research related to NIMH Division of Services and Intervention Research. The aim is to design and test the first prototype of an interactive self-help computer program for binge eating disorder. The prototype will be based on an integrative view of the etiology of binge eating disorder, and will combine elements of various empirically supported treatment approaches. Each user will receive a customized learning experience, resulting in customized feedback from the program. We will conduct the following evaluations of the module: (1) Expert Reviews of Content and Usability, (2) Stage One Usability Test (with eating disorder subjects), and (3) Stage Two Usability Test (with eating disorder subjects). Dependent measures and formal criteria for success are specified. PROPOSED COMMERCIAL APPLICATION: Potential commercial markets for eating disorders self-help technology include psychotherapy clients, mental health professionals, and health maintenance organizations.


The psychoactive component of marijuana, ?9-tetrahydrocannabinol, binds to and activates cell surface receptors called cannabinoid receptors. Activation of these receptors produces intense responses in humans, which suggest that endogenous cannabinoid (endocannabinoid) substances may contribute in important ways to brain functions as diverse as appetite, mood and pain. Two such substances are anandamide and 2- arachidonoylglycerol (2-AG), lipid-derived compounds that are released from neural cells and activate cannabinoid receptors with high affinity. Anandamide and 2-AG undergo a rapid deactivation process, which contributes to arrest of their biological actions. In the case of anandamide, this process is thought to consist of two steps, transport into cells and intracellular hydrolysis by the enzyme fatty-acid amide hydrolase (FAAH). By contrast, much less is known about 2-AG deactivation. Previous work in our lab has provided evidence that the enzyme monoacylglycerol lipase (MGL) - a presynaptic serine hydrolase that converts 2-AG into arachidonic acid and glycerol - participates in 2-AG hydrolysis. Based on these results, we hypothesize that MGL is an essential component of the physiological mechanism by which 2-AG is deactivated at brain synapses. Our first objective is to discover potent and selective inhibitors of MGL activity. In our initial studies, we have identified two promising leads, which will be optimized through a series of experimental and computational approaches, including structure-activity relationships, site-directed mutagenesis, mass spectrometry and molecular modeling. Our second objective is to characterize 2-AG transport into neurons and define the role of MGL in this process. We will combine pharmacological and genetic strategies to test the hypothesis that MGL- mediated hydrolysis provides the driving force for neuronal 2-AG internalization. Finally, we will use MGL inhibitors and mutant mice that overexpress MGL in the forebrain to determine whether 2-AG hydrolysis terminates the behavioral effects of endogenous 2-AG. We will focus our attention on two behaviors - feeding and stress-coping responses - in which as-yet-unidentified endocannabinoid signals are thought to play a crucial role. These studies will help uncover the functions served by the endocannabinoid system in the brain and may open innovative avenues for the treatment of neuropsychiatric and substance abuse disorders. PUBLIC HEALTH RELEVANCE: Marijuana works by mimicking important transmitter substances in the brain, called endocannabinoids. The objective of our research is to understand how these transmitters are produced and eliminated, and discover innovative medicines that target these processes.


DESCRIPTION: The Hsp70 chaperone system of E. coli consists of the Hsp70 homologue DnaK, the Hsp40 homologue DnaJ, and the Hsp20 homologue GrpE. The coordinated action of these proteins is important in the folding of some proteins and in regulating the action of several macromolecular assemblies. This proposal focuses on the DnaJ component of this chaperone system. DnaJ is a co-chaperone that forms complexes with DnaK and also with substrate proteins. It contains four evolutionarily conserved, functionally distinct domains or modules. The structure of individual modules of DnaJ will be determined using 13C/15N/1H heteromuclear multidimensional NMR spectrosocpy, and residues that are important for function will be identified by site-directed mutagenesis. Complexes between modules of DnaJ and fragments or domains of its various protein partners will be characterized by NMR spectroscpy. These studies will provide insight into the roles of DnaJ and its homolgues in this important chaperone system.


Glioblastoma multiforme (GBM) is the most common malignant glioma and has an extremely invasive phenotype. The ability of glioblastoma cells to diffusely infiltrate healthy tissue is the major obstacle for current treatments, including surgery and radiotherapy. Over the years, established human glioblastoma cell lines were used as models to study molecular mechanisms of invasion; however, these mechanisms remain elusive since established lines are not invasive in vivo. In contrast to established lines, primary non-established glioblastomas, including GBM6 and GBM12 cells, are invasive in vivo, and thus can be successfully used to study growth, motility, and invasion. Recently, an elevated expression of sphingosine kinase 1 (SphK1), an enzyme that generates sphingosine 1-phosphate (S1P), was correlated with poor prognosis of patients with GBM. S1P is a potent lipid mediator important for cancer growth, survival, migration, and invasion. In the brain, S1P is present at high concentrations, and has been shown to stimulate migration and invasion of established glioblastoma lines in vitro. The activity of SphK1 is rapidly and transiently stimulated by various growth factors and cytokines; however, mechanisms of the transcriptional regulation of the sphk1 gene are not understood and the factors enhancing SphK1 expression in GBM patients have not been identified. Recently, inflammation has been linked to the development and progression of many cancers with inflammatory cytokines involved in these processes. IL-1 is one of the major regulators of inflammation in the brain, which can induce the secretion of other proinflammatory cytokines and also promote proliferation of glioblastoma cells in vitro. More importantly, glioblastoma cells secrete significant amounts of IL-1. In preliminary studies, we show that IL-1 is coexpressed with SphK1 in GBMs, exogenous IL-1 increases expression of SphK1, and both IL-1 and S1P enhance invasion and proliferation of glioblastoma cells. More importantly, neutralizing anti-IL-1 antibodies inhibit proliferation of glioblastoma cells in vitro, while a novel SphK1 inhibitor (SKI) reduces xenografted glioblastoma tumor growth in mice. It is our hypothesis that the IL-1-mediated stimulation of SphK1 expression leads to the production of S1P, which in turn activates proliferation, motility, and invasion of GBMs. We propose to study SphK1 functions and its regulation in primary GBM cells. Moreover, development of potent and specific SphK1 inhibitors that are active in animal models could pave the way for new therapeutics for treatment of GBM. Our specific aims are as follows: Aim 1. To determine the role of SphK1 expression and the effect of its inhibition on proliferation, motility, and invasion of primary non-established GBMs in vitro, and xenografts implanted in the brains of athymic mice. Aim 2. To identify the mechanisms that control endogenous SphK1 expression in response to IL-1. PUBLIC HEALTH RELEVANCE: Glioblastoma multiforme (GBM) has an extremely invasive phenotype, which is the major obstacle for current treatments. It is our hypothesis that the IL-1-mediated stimulation of SphK1 (sphingosine kinase 1) expression plays an important role in GBM growth and invasiveness, and may be a potential therapeutic target. Therefore, we propose to study SphK1 functions and its regulation in primary non-established GBM cells. Moreover, development of potent and specific SphK1 inhibitors that are active in animal models could pave the way for new therapeutics for treatment of GBM. [unreadable] [unreadable] [unreadable] [unreadable]


Several hypotheses have been created to account for the neurodegeneration and subsequent cognitive deficits observed in Alzheimer's disease. One hypothesis, in particular, has focused on the effects of inflammation as a mediator of neurodegeneration. To address this possibility, we have initiated studies examining the direct and indirect effects that endotoxin and inflammatory cytokines may have on neural tissue and neuronal cell signaling. Initial studies have demonstrated that the intracerebral infusion of endotoxin produces a significant age-related increase in brain tumor necrosis factor- alpha (TNFa) levels, but does not effect the production of a number of other inflammatory cytokines such as interleukin-1 or interleukin-12. We are currently examining young and old rodent brain homogenates and plasma post endotoxin treatment for the presence of various inflammatory cytokines (e.g., interleukin-6, interleukin-4, interleukin-10) and chemokines (e.g., interleukin- 8, MCP-1). Moreover,animals are also being examined for the effects of direct cytokine administration on inducing brain inflammation, permeabilizing of the blood-brain barrier, and effects on leukocyte trafficking, CNS surface markers, neurodegeneration, and cognitive behavior. We believe that cytokine and chemokine infusion (icv and iv) in rodent brains will have significant biological and physiological effects on neural tissue and will ultimately reveal a relationship between neuroinflammation, age, and cognitive behavior. These studies have been initiated using intracerebral TNF-a infusion and we hope to soon follow these efforts with chemokine and interferon infusions. In addition, additional studies will also be performed to determine whether administration of signaling inhibitors and antagonists and/or through in vivo leukocyte depletion can ameliorate the biological and physiological effects of administered cytokines. It is our hope that these efforts will assist in our understanding of the contribution of cytokines and chemokines in the neuroinflammatory and neurodegenerative effects observed in various neurotrauma models and age-related disease states.


This proposed Research Center in Practice-Based Research is a collaboration of six well-established practice-based research networks (PBRNs) that have joined together to create the Meta-network Learning and Research Center (Meta-LARC.) Meta-LARC's mission is to sustain a consortium of practice-based research networks dedicated to increasing the quality, effectiveness and safety of primary care through accelerated research and collaborative learning. Meta-LARC networks include the Oregon Rural Practice-based Research Network (ORPRN, home of the new Center), the lowa Research Network, State Networks of Colorado Ambulatory Practices and Partners, Oregon's Safety Net West Practice-based Research Network, the Quebec Practice Based Research Network, and the Wisconsin Research and Education Network. These PBRNs are comprised of 533 primary care practices and over 6,000 clinicians who provide care for an estimated three million patients in rural, urban, and underserved communities. Combined, the networks have conducted over 200 studies and published nearly 200 scientific publications. The specific aims of Meta-LARC are described below: Aim 1: Foster the capabilities of six PBRNs and 533 primary care practices through a robust collaboration designed to conduct research to improve the quality, effectiveness and safety of primary care. Aim 2: Accelerate the conduct of PBRN research through a well designed, high functioning common infrastructure that enables the efficient conduct of research. Aim 3: Promote continuous learning and sharing across Meta-LARC networks and practices to accelerate the dissemination of knowledge and bi-directional communication.


The 37 million Americans who live in poverty smoke at twice the rate of other Americans. As a result, those living in poverty bear a disproportionate burden from tobacco-related diseases, including lung cancer, other lung diseases, and heart disease. A number of beliefs serve as barriers to their use of evidence-based treatment for tobacco dependence. These beliefs include: 1) smoking is both normative and acceptable under some conditions;2) willpower is sufficient for quitting rendering outside help unnecessary and irrelevant;3) evidenced-based treatments are not more effective than other methods;4) quitting medicines are ineffective, dangerous, addicting, and/or too expensive;and 5) treatments for quitting are not available, hard to access, and/or too expensive. Also, while the poor have as much desire to quit as others, their motivation to make a quit attempt in the near term is less than that of other smokers. New ways are needed to bring evidence-based treatment to this hard to reach population. Providing a brief tobacco dependence intervention to the poor seeking services from community agencies is one such way. Specifically, this study will examine the effectiveness of a brief cognitive-motivational intervention designed to challenge these beliefs and motivate a quit attempt by smokers not otherwise motivated to quit who are presenting for services at the Salvation Army (N=140) relative to an attention control group (N=140), a no-treatment control group (N=140), and smokers who are motivated to quit (N=100). The primary outcome is acceptance of a brief treatment intervention. Other outcomes are subsequent quit attempts and abstinence through three months of follow up. In addition, community agency staff who administer the interventions will complete a survey to assess its feasibility in the community agency setting. Data will be analyzed to evaluate study aims: 1) Did the brief cognitive-motivational intervention result in greater acceptance of treatment, quit attempts using evidenced-based methods, and abstinence than the control groups? 2) Did the intervention boost treatment acceptance and quit attempts above the rate by smokers already motivated to quit? 3) Is the intervention feasible for use in community agencies and acceptable to its cliental? If these aims are achieved, a brief, practical, and effective intervention can be available from community agencies that motivates smokers living in poverty to make quit attempts using evidenced-based methods. Wide distribution of this tailored intervention will help decrease the health disparity experienced by smokers living in poverty. PUBLIC HEALTH RELEVANCE: People living in poverty are far more likely to smoke than others and, as a result, have more tobacco-related disease. The poor are not as motivated to quit and have certain beliefs that prevent them from using the best methods of quitting when they do try. This research will determine if a brief intervention designed to motivate and change these beliefs increases quit attempts when it is provided by a community agency that serves the poor. If successful, this research will help reduce tobacco-related diseases among the poor.


Family caregivers to elders suffering with dementia are at increased risk for stress-related illness. This is especially true for Latino caregivers, whose burden is exacerbated by a lack of Spanish-language information on dementia, and/or culturally-competent services. Even when such services are available, Latino caregivers often face access barriers related to scheduling convenience, geographic distances, and the inability to leave their dementia-affected elder unattended. This project addresses the need for convenient, culturally-competent, Spanish-language services for Latino caregivers. We propose the development of a multi-literacy level Spanish-English website for family caregivers to dementia-affected elders. We have already established a prototype website for Latino caregivers in the Boston are, and will further develop it in Phase 1. We will do this by expanding the higher literacy Spanish component of the website, as well as developing lower literacy versions in Spanish and English. An evaluation study of the website as a learning tool will be conducted. In Phase II, we will expand the website to include caregiver resources nationally, and add a customized e-care component offering on-line and call-in center advice from dementia specialists. HMOs can be expected to contract with us to provide the e-care component as a member service.


PROJECT SUMMARY: Project 3 The major goal of Project 3 is to provide a detailed structural and mechanistic understanding of human norovirus (NoV) evolution and replication to aid in the design and development antiviral strategies. Human NoVs are the leading cause of epidemic acute gastroenteritis. Susceptibility to these viruses is determined by genetically controlled expression of histo-blood group antigens (HBGAs), which are also critical for NoV attachment to host cells. As a result of sequence changes in the P domain of the capsid protein VP1 that harbors the HBGA binding site, these viruses show strain-dependent variability in HBGA specificity presenting a fascinating case study in how genotypic variations allow for exploitation of the polymorphic nature of HBGAs in host populations to counter herd immunity and cause epidemics. Recent studies have shown human antibodies that block HBGA binding correlate with protection from NoV-associated illness (Project 1). Several studies have suggested a correlated interplay between antigenicity and HBGA specificity in driving the evolution of NoVs. The goal of AIM 1 is to provide the structural basis for such interplay by structurally characterizing how HBGA-blocking antibodies interact with NoV strains using currently available human monoclonal antibodies and those newly developed in Project 1, using X-ray crystallography and cryo-EM techniques. While the emphasis in AIM 1 is on virus-host interactions, the emphasis in our following two AIMs is on two key virus encoded proteins that regulate virus replication, both of which are potential targets for small molecule drug discovery. AIM 2 focuses on the viral protease that is critical for polyprotein processing. Based on a recent exciting finding by us and others that NoV protease binds to viral RNA, we have hypothesized that it plays a hitherto uncharacterized role in genome replication. In the first part of AIM 2, our goal is to determine the structural basis of genogroup-dependent substrate interactions and inhibition in order to provide a rational framework for the design and optimization of inhibitors in collaboration with Project 1. In the second part of this aim, our goal is to structurally characterize NoV protease interaction with viral RNA, and in collaboration with Project 2, probe into the role of this interaction in viral replication. AIM 3 will focus on Norwalk virus NTPase, p41, a member of a highly conserved family of proteins encoded by a wide variety of (+)RNA viruses that plays a critical role in virus replication by remodeling cellular membranes into vesicular compartments in infected cells. During the current grant period, we provided the first structural characterization of such a protein from our studies on p41. The goal of this aim is to take the next step of addressing mechanism-related questions using a combination of crystallography, cryo-EM and cell-based techniques and to correlate our observations to the functional aspects of this protein during virus replication in collaboration with Project 2. We expect our proposed studies will provide novel structural and mechanistic insight that will have significant impact on immunological, translational, and replication-related aspects of human NoVs.


Lung injury associated with neonatal Respiratory Distress Syndrome and Bronchopulmonary Dysplasia requires understanding developmental responses of pulmonary proteolytic activity and defense mechanisms intrinsic to the newborn lung. Utilizing neonatal pulmonary effluent obtained by sequential tracheal aspiration, the aims of this proposal are to: 1. Determine sequentially, the levels of protease activity and protease inhibition in pulmonary effluent of human preterm infants and to relate these biochemical indices of lung injury to radiographic and ventilatory assessments to the severity of acute abd chronic lung disease, 2. Using homologous surface active materials derived from term human amniotic fluid, the proposed investigations seek to define whether surfactant supplementation in 20-30 preterm, infants per year with respiratory distress syndrome alters the clinical, radiological, cytologic, proteolytic activity, and protease inhibition activity in infants treated with surface active material supplemention in comparison to infants receiving conventional mechanical ventilatory support with oxygen supplementation using a multiple logistic regression model. In addition, these investigations will define whether or not evidence of immune complex formation results in infants receiving homologous surface active material using conglutinin assays, as well as, measurement of anti-surfactant antibody formation in the sera of treated infants throughout the first year of life in order to determine whether detectable immunologic reactivity occurs from tracheally instilled surface active material in developing preterm infants, 3. To determine whether active Alpha1 protease inhibitor levels, elastase levels, and alastase inhibitory capacities are altered in neonatal rabbits in graded exposure to varying concentrations of FiO2 previously established to result in lung injury, 4. Using neonatal and young New Zealand White rabbits, describe the effects of graded exposure of intratracheally instilled human derived surfactant on lung histology, presence of circulating antigen, and measures of pulmonary immunoreactivity.


The nature and function of metal atoms in enzymes and proteins has been a keystone of biomedical research since Warburg's initial discoveries in the 1930's on "heavy-metal enzyme catalysis" and Keilin's identification of hemoproteins. Many biophysical approaches to the study of these heavy metal enzymes have been in process. Optical and electron paramagnetic resonance studies are good examples of those that give definitive information on many properties but have "blind spots" that fail to give incisive information on the metal atoms in all valences or states, i.e., certain properties are "invisible" to most techniques. K edge absorption of X-rays by heavy metal atoms occurs in all valence states and gives definitive fingerprints of the nature of such atoms and their electronic configuration and chemical environment. Extended edge absorption studies (exafs) give many other properties, some of which can be interpreted as precise distance measurements to neighbors of the heavy metal atoms. While related data may ultmately be inferred from x-ray or nuclear magnetic resonance data, the great potential advantages of the exafs method are the possibilities of structural studies of the metalloproteins that are time resolved at enzymatically functional concentraions. Optical and epr sample monitoring detects any change of valence or liganding under synchrotron irradiation. This proposal seeks to set up instrumentation development and testing of components that increase the efficiency of photon collection from x-irradiated samples, the ultimate goal being to achieve edge absorption EXAFS and anomalous scattering studies in the range appropriate enzyme activity studies, i.e., less than 100 microns M and eventually extending down toward micromolar concentrations. Concomitant with this development are essential innovations necessary for kinetic studies of the state of mental atoms in enzymatic function. Last but not least are improved methods for on-line sample monitoring to ensure the integrity of the biological material throughout the course of the x-irradiation.


There is growing recognition of the importance of 'place' on individual risk of disease, although most studies fail to capture the dynamic nature of the risk environment and its effect on the sex trade has been understudied. The Mexico/US border region is undergoing profound alterations in the environment in which sex work takes place, providing a 'natural experiment' through which we can explore the relative contributions of individual and structural factors on HIV transmission. The primary goals of this research are to define social, spatial, and physical factors affecting female sex workers (FSWs) in 2 border cities and determine their relevance to HIV epidemiology, drug use, and access to services. Tijuana and Ciudad (Cd.) Juarez have witnessed escalating community-level violence and, for Cd. Juarez, changes in the location and visibility of the 'red light' district. While migration and cross-border interactions are major influences, the recent global economic downturn is dramatically altering migration patterns. Based on the above, the specific aims of this project are to: 1) assess changes in social influences on the sex work risk environment over time in both cities and their effect on risk behaviors, HIV/STI incidence, and access to services; 2) determine the locations where FSWs live, work and engage in other activities and their relationship to risk behaviors, perceptions of violence, and access to services; and 3) determine the extent to which the built environment and other sex work venue characteristics relate to individual-level behaviors and HIV/STI incidence. To meet these aims, we will recruit 600 FSWs (300 per city) and collect sociodemographic, location, and behavioral data through interviews. All will be tested for HIV, syphilis, gonorrhea, and Chlamydia and treated as needed. Follow-up interviews and testing will occur at months 6, 12, and 18. To meet aim 2, we will construct a geographic information system (GIS) of both cities and explore factors in relation to where FSWs live, work, buy/use drugs, and access services. Changing spatial relationships, such as the dispersal of the Cd. Juarez Zona Roja and intra-urban or cross-border mobility, will be analyzed to track patterns of infectious disease spread. We will also conduct in-depth interviews and an activity-travel survey with 30 sex workers per city, stratified by geography and venue (e.g. street, bar, etc.) to create geo-narratives based on time-geographic methods and computer-aided qualitative data analysis in order to explore the impact of recent social, economic, political, and other structural changes on participants' lives. To meet aim 3, field measurements of the built environment and other venue characteristics will be combined with individual-level data to explore their effect on health outcomes. The data collected will provide information vital to reframing HIV and drug use interventions to take into account the structural environment. This project is timely as HIV prevalence is rising along the U.S./Mexico border, presenting a window of opportunity to prevent transition to a generalized epidemic.


The purpose of this trial is to determine if there is a difference between different routes of intermittently administered rhIL-2 (IV vs subcutaneous admininistration) with a highly active antiretroviral regimen compared to highly active antiretroviral therapy alone with respect to the proportion of subjects achieving a 50% increase in CD4 counts above pre-randomization baseline values after one year of rhIL-2 and the rate of change in CD4 counts. Another primary hypothesis will focus on safety, tolerance, and effect on quality of life of these regimens. A third aim will focus on changes in immune cell phenotypes and function and on HIV viral load and rate of antiviral drug resistance development when HAART is used in the setting of rhIL-2 compared to HAART alone. The ACTG 872 Substudy will examine specific and different immunologic effects when comparing the two regimens of rhIL-2 plus HAART vs. HAART alone. The ACTG 874 substudy will determine the cellular source of HIV production in lymph node tissue before and after treatment with HAART, and the changes induced in lymph node tissue with the addition of exogenous rhIL-2, in a subset of subjects (n=12). This is a randomized study to ascertain the effect on CD4 response of high dose, intermittently administered rhIL-2 administered by continuous intravenous infusion versus subcutaneous injections in conjunction with highly active antiretroviral therapy versus highly active antiretroviral therapy alone in patients with HIV and CD4 counts of 50-300 cells/mm3. Subjects must be protease inhibitor therapy naive. All subjects will receive treatment with indinavir and 2 nucleoside analogues; one of which is new to the subject (HAART). Subjects will receive 12 weeks of HAART. At week 10 an HIV RNA by bDNA (real time) will be done by Chiron. Subjects with < 5,000 copies/ml will be continued on study. Subjects with > 5,000 copies/ml will require no further follow-up and should be taken off study. Subjects who have < 5,000 copies/ml of HIV RNA will then be randomized to continue HAART alone or HAART with rhIL-2 at either 9 million units by continuous infusion (CIV) for 5 days every 8 weeks for 72 weeks, or 9 cycles, or rhIL-2 at 7.5 million units by subcutaneous injection twice a day for 5 days every 8 weeks for 72 weeks or 9 cycles. Subjects on the CIV rhIL-2 arm who achieve both a $ 25% increase and at least a 100 cell/mm3 increase in CD4 count above the prerandomization baseline value after 3 cycles or after 6 cycles will switch to rhIL-2 at 7.5 MIU by subcutaneous injection twice a day for 5 days every 8 weeks for the remainder of their treatment course.


The long-term goal of the candidate is to be an independent investigator conducting interdisciplinary studies on neural correlates of substance use disorders (SUDs), especially brain-based predictors of treatment outcomes of SUDs by integrating neuroimaging and clinical trial methods. Brain-based predictors have the potential to guide the optimal patient-therapy matching to improve the efficacy of treatment for SUDs. His short-term goal is to gain required skills for achieving his long-term goal through the training supported by this K01 Award. The candidate received training in Medicine and Diagnostic Radiology in China, a Ph.D. in Neuroscience from the Medical College of Ohio, and T32 Postdoctoral training in functional magnetic resonance imaging (fMRI) on SUDs from the University of California Los Angeles (UCLA). Since 2007, the candidate has been a faculty level investigator in the Division of Substance Abuse at the Yale School of Medicine Department of Psychiatry. His research has focused on using fMRI to identify neural correlates of treatment outcomes of cocaine dependence. The candidate now seeks support to expand the scope of his research on neural predictors of treatment outcomes of other SUDs and to obtain additional training focused on mastering diffusion tensor imaging (DTI) analysis and on learning principles of SUD treatment and clinical trial methods. His preliminary findings indicate that better integrity of white matter in the frontal lobes and anterior corpus callosum positively correlates with better treatment outcomes of cognitive behavioral therapy for cocaine dependence. The candidate seeks to replicate and expand these initial findings in studies that use different therapies and with patients of other SUDs. His research strategy will be to conduct a series of analyses of fMRI, DTI, and treatment outcome data to identify neural predictors and correlates of treatment outcomes of patients dependent on cocaine or other substances participating in 5 clinical trials of several different therapies for SUDs. His training program will cover the following topics: 1) enhancing and expanding his expertise in neuroimaging, 2) learning clinical trial methodologies for SUDs, particularly as they relate to brain measures, 3) sharpening interdisciplinary thinking about brain correlates of addiction treatments, 4) enhancing statistical knowledge for longitudinal research, 5) enhancing knowledge on ethics related to human studies and responsible conduct of research, and 6) training in preparation for manuscripts and grant proposals. His mentors include internationally prominent experts in neuroimaging (Marc Potenza, M.D., Ph.D., Godfrey Pearlson, M.D.) and in SUD treatment research (Bruce Rounsaville, M.D., Kathleen Carroll, Ph.D.).


Filipinos in Hawaii have high rates of end-stage renal failure and are disproportionately represented on the local transplant waiting list. The goal of the proposed project is to increase cadaveric organ, tissue and eye donation by Filipinos in Hawaii through a comprehensive statewide education program. The project's alms are to increase donor card signing and family discussions among Hawaii Filipinos as well as to increase the number of cadaveric organ donors to 35% of Hawaii donors. The project will partner with Filipino health, social, cultural and religious organizations and will use interviews, review of records, focus groups and surveys to: identify why Filipinos have historically low rates of donation; evaluate Hawaii's previous Filipino education program (MOTTEP of Honolulu); and develop minority-targeted messages delivered by ethnically and culturally-similar and sensitive messengers. The messages will emphasize use of personal experiences, trusted community leaders and mechanisms such as informal networks and Filipino media. Filipino health care workers will also be trained to assist the local organ procurement organization in offering the option of donation to Filipino families of potential organ donors. Because Filipinos will soon comprise the largest Asian-American ethnic group in the United States, Bayanihan may serve as the model for other under-represented minority ethnic groups throughout the United States in the future.


Introduction: The events of September 11, 2001, and the deliberate bioterrorism attack with anthrax that followed, led to profound changes in the biologic research and public health agenda of the United States. The recognition that the United States was vulnerable to international terrorism, and the tremendous impact that a relatively limited biologic weapons attack had on the nation, resulted in major changes in the research focus of government, academic, and commercial entities. Ml AID understood that the United States lacked the research capacity and infrastructure to respond to the threat posed by biological weapons and emerging infectious diseases. The Regional Centers for Excellence in Biodefense and Emerging Infectious Diseases Research (RCE) program was a central component of the NIAID response to this threat, and was designed to "jumpstart" research in biodefense within each of the 10 public health service regions of the U.S. In 2003, Washington University in Saint Louis, Saint Louis University, the University of Missouri, Case Western Reserve University, and the Midwest Research Institute received funding to establish the Midwest Regional Center for Excellence in Biodefense and Emerging Infectious Diseases Research (MRCE) to support research on biodefense in Region VII (Iowa, Kansas, Missouri and Nebraska) and parts of Ohio. The goals of the MRCE were consonant with the goals of the RCE program: harness the best scientists in the region for discovery and translational research designed to lead to the next generation of therapeutics, diagnostics and vaccines for biological threats;provide training and career support to create a new generation of scientists working in this field;establish core facilities to support researchers throughout the region;and develop an emergency response plan that can rapidly bring to bear our scientific expertise and research capacity in the event of any new biological threat to the region, the nation or the world. Over the past 5 years the MRCE has made significant progress towards each of those goals, and we will detail our accomplishments in subsequent sections. Given the scope of the program, and our aspirations, it is not surprising that much remains to be done. While the fundamental goals of the MRCE remain the same, this competitive renewal is built upon a research landscape that has been transformed by the RCE program. When we prepared the MRCE proposal 5 years ago, there was strength in basic immunology and in microbial pathogenesis research at the partner institutions, but relatively little was focused on the NIAID priority pathogens. Research collaborations between the MRCE partner institutions did not exist on any substantive level, and even intra-institutional collaborations in this research area were limited. There was no pipeline of young scientists and clinicians interested in research careers in biodefense, and there were no established biosafety training programs within the region. The MRCE now coordinates and supports a vibrant, highly collaborative and closely integrated group that has become the face of biodefense and emerging infectious diseases research throughout most of Region VII. MRCE programs have created a pool of talented young investigators interested in biodefense and emerging infectious diseases research. We have trained nearly 100 individuals to work safely at BSL3, and have educated investigators and public health officials throughout the country. The MRCE has responded to three separate regional emergencies, and has become an integral part of the disaster planning process for a significant part of our region. We have more than doubled the number of participating institutions, and have supported researchers from 10 different academic or industrial organizations. Most importantly, MRCE investigators made seminal basic science and translational discoveries leading to new therapeutic targets for West Nile, Dengue fever, pneumonic plague, Ebola, poxviruses, and a wide sector of RNA and DNA viruses (section A.3). We also initiated a pioneering collaborative effort in pathogen discovery and assessment that led to the recognition of 7 new viruses, one of which (WU polyomavirus), may be an important cause of respiratory disease in children. In our original application 5 years ago, we discussed how one could best assess the impact of a complex and wide ranging program like the MRCE on the research effort in Region VII. We proposed then that the most objective measure would be the change in extramural funding for biodefense and EID research in Region VII (excluding MRCE funds) during the course of the MRCE program. Since 2002, (the baseline pre-MRCE year) total extramural funding for biodefense and EID research (excluding MRCE and regional laboratory allocations) in Region VII has increased more than 2.5 fold, with the greatest increases in 2006 and 2007 despite a flat NIH budget. We believe the MRCE program has had a significant impact on Region VII and the nation, and are committed to continuing this important work.


The active constituents in allergenic extracts of Poison Ivy and other plants of the genus toxicodendron have been generically designated as "urushiol", a mixture of penta-or heptadecenyl catechols. Objectives of this project are: a) to develop methods for the quantitative analysis of urushiol, including speciation of individual compounds; b) to develop sample preparation techniques to isolate urushiol from plant oil matrices; c) to develop methodologies to determine oxidized and oligomerized urushiol degradation products; d) to correlate urushiol degradation with factors of concentration, solution type and storage conditions. Urushiol samples have been speciated and quantitated by gas chromatography of trimethylsilyl derivatives. Validation studies have been performed for this method. A normal phase HPLC method utilizing a silica column is to be investigated. Development of a capillary gas chromatographic procedure, providing better resolution of individual compounds is in progress. Procedures for the isolation of urushiol at high (greater than 100 ppm) concentrations from plant oils have been validated.


This proposal requests support for a Keystone Symposia meeting entitled Immunity in the Respiratory Tract: Challenges of the Lung Environment, organized by David L. Woodland, Liise-anne Pirofski and Allen G. Harmsen, which will be held in Vancouver, British Columbia, Canada from February 26 - March 3, 2011. Recently, there has been an explosion in information about the regulation of inflammation, immunity, and immunopathology in the lung. Despite these advances, we still have only a rudimentary understanding of how these responses relate to the lung environment or how and why they translate into beneficial or detrimental effects, health or disease. This meeting will bring together allergists, immunologists, microbiologists and vaccinologists to discuss pulmonary inflammation and immunity in the context of lung biology, and to build on this understanding to promote development of improved vaccines and therapeutic interventions for inflammatory conditions and infectious diseases that affect the lungs. Expert talks in plenary sessions will present the latest research in the field. We anticipate that the meeting will considerably advance our understanding of respiratory immunity. Moreover, opportunities for interdisciplinary interactions will be significantly enhanced by the concurrent meeting on Mucosal Biology: A Fine Balance between Tolerance and Immunity, which will share a keynote address and two plenary sessions with this meeting. Project Narrative: Pneumonia is a leading killer of children worldwide under the age of five years, as well as an important cause of morbidity and mortality in adults in the developed and developing world. In light of this, the importance of the Keystone Symposia meeting on Immunity in the Respiratory Tract: Challenges of the Lung Environment cannot be overestimated: leading scientists from diverse backgrounds and settings will engage in critical discussions about the disease and challenges to understanding immunity in the lung and controlling lung inflammation.


Reconstituted high-density lipoprotein (R-HDL) has been shown in vitro and in vivo to have antiendotoxin properties. The hypothesis is that lipoproteins bind, neutralize, and increase clearance of endotoxin. Being a natural biological product, R-HDL could be used prophylactically to prevent septic shock in high-risk patients and in chemotherapy-induced neutropenia, or therapeutically to treat septic shock. To test this hypothesis, in a controlled, randomized trial, we investigated the effects of R-HDL on survival, endotoxemia, cytokine production, and pathophysiologic and metabolic events in our canine model of septic shock. At 0.5, 8, and 16 h after implantation of an E. coli-infected clot, canines received intravenous R-HDL control lipid, or human serum albumin (HSA). Animals treated with R-HDL had lower levels of circulating endotoxin and tumor necrosis factor (TNF) and a smaller decrease in white blood cell counts than did animals treated with lipids and HSA. Survival times of lipid-and HSA-treated animals were similar and were significantly greater than those of R-HDL-treated animals. R-HDL-treated animals had significantly greater abnormalities in liver function compared to lipid- and HSA-treated animals. In normal animals, R-HDL caused transient elevation of liver enzymes. In animals given sterile clot intraperitoneally, R-HDL caused seizures. Thus, this preparation of R-HDL had antiendotoxin effects, but also had hepatic and neurologic toxicities. We now plan to investigate a different preparation of HDL that has a different chemical composition than that used in this canine experiment. This new preparation was manufactured so as to limit the potential of liver and neurological toxicity. First, we will investigate the new R-HDL preparation in small animal models of live-infection (rats), and subsequently, if no toxicity and beneficial antiendotoxin effects are documented, in our canine model of septic shock. If toxicities associated with R-HDL can be reduced, its antiendotoxin effects may potentially be used prophylactically in high-risk patients, or therapeutically in patients with established septic shock.


Research project CA-03641 deals with pathways and rates of hydrogen flow in intracellular oxidation-reduction reactions. Our studies are based on the concept that, in intact cells, pathways of hydrogen are determined not only by thermodynamic properties of the interacting systems but also by factors which influence the kinetics, such as heterogeneity of distribution of dehydrogenases within intracellular compartments, specific enzyme-enzyme interactions, and the different strength of binding of the pyridine nucleotide coenzymes by dehydrogenases. A primary objective of these studies is to attempt to locate steps which control the rates of biological oxidations in intact cells, so that it may be determined whether control of the reductive biosynthesis of cellular constituents, i.e. fatty acids, cholesterol, etc. occurs at the level of the obligatory oxidation-reduction reactions which are an essential part of the biosynthetic process.


Metastatic melanoma is a deadly disease with an expected five-year survival of only 14%. The goal of this project is to better understand the environmental and genetic factors which cause this often-fatal disease in the hope that we may suggest approaches for disease prevention and therapeutic intervention Epidemiology has suggested that ultraviolet (UV) radiation and the red/blond, fair-skinned pigmentation phenotype are two major melanoma risk factors. However, genetically controlled experiments to establish an unambiguous causative link is lacking. This project aims to create a series of mouse models for two main objectives: (1) to establish a clear link between UV, pigmentation and melanoma in order to provide better public health information, and (2) to use these mouse models to understand the molecular mechanisms by which UV and high risk pigmentation variants produce melanoma. In order to produce a novel melanoma mouse model, red/blond, albino, and black mice which also carry an activatable form of BRAF or NRAS (the most commonly mutated oncogenes in melanoma) are being generated. Preliminary experiments using this system have already revealed the striking observation that a significant number of the red/blond mice develop melanomas, while none of the albino mice and very few of the black mice develop the same lesions. With this data suggesting there is a strong genetic tumorogenic effect of the red/blond phenotype, it is likely that examining the role of UV will provide exciting new information. To pursue the first aim, melanoma rates between UV irradiated red/blond, albino, and black mice will be compared to determine the effects of pigmentation and UV alone. Next, melanoma rates between UV irradiated black mice that carry either of the 2 most commonly mutated oncogenes in melanoma (BRAF or NRAS) will be compared to determine the effects of oncogene activation and UV irradiation. Lastly, melanoma rates after UV irradiation of red/blond and albino mice carrying the same mutated oncogenes will be compared to see how the 3 variables work together to alter melanoma risk. The goal of the second aim is to understand the mechanisms by which UV and the red/blond phenotpye induce disease. Initially it will be investigated if the high rate of melanoma in the red/blond context is pigmentation dependent. Next, it will be investigated if the mechanism of red/blond-melanoma is via reactive oxidative damage, as suggested by previous studies showing that red/blond pigment can promote release of reactive oxidative species (ROS). To investigate this question, the types of DNA damage in UV irradiated red/blond, albino, and black mice will be compared. Finally, the ROS burden in melanocytes will be genetically and pharmacologically altered to investigate if this strategy can block red/blond-melanoma induction. PUBLIC HEALTH RELEVANCE: Metastatic melanoma is a deadly disease with an expected five-year survival of only 14%. Alarmingly, in the United States, the incidence of melanoma is rising at a faster rate than any other malignancy. The goal of this project is to better understand the environmental and genetic factors that contribute to development of this often-fatal disease, in the hope that we may use this knowledge to suggest approaches for disease prevention and nodal points for therapeutic intervention


This Mentored Patient-Oriented Research Career Development Award proposes a program of tailored mentorship, focused research, didactics and coursework to provide the candidate with the training in sleep medicine and perinatal neuroendocrine functioning necessary for independence as an investigator. Candidate. The candidate's background and foundation in clinical research make her well-poised for this award. She completed her Ph.D. in clinical psychology at the University of Alabama, where she received training rooted in the scientist-practitioner model. An emerging interest in sleep and mood led her to pursue a postdoctoral fellowship at the University of Michigan, where she began to develop a niche in behavioral sleep medicine and perinatal mental health. She was recently appointed to the faculty of the Department of Psychiatry at the University of Michigan. Her goal is to receive additional training in sleep medicine and perinatal neuroendocrinology. The candidate has identified two key training objectives necessary to achieve full independence as a researcher: (1) experience in sleep physiology (including assessment and treatment of sleep-disordered breathing in pregnancy) and (2) neuroendocrinology in pregnancy. Environment. The proposed project will take place within the University of Michigan Department of Psychiatry and the Sleep Disorders Center. Both departments have an extensive history of close collaboration with the Department of Obstetrics and Gynecology. The fifth largest research university in the United States, the University of Michigan provides extensive resources and a rich research environment. The candidate will have access to the large, accredited Sleep Laboratory within the Sleep Disorders Center, the well-established perinatal recruitment infrastructure within the Women's Mental Health and Infants Program (in collaboration with the Department of Obstetrics), and resources provided by the CTSA-funded Michigan Institute for Clinical and Health Research. The proposed primary mentor, Ronald Chervin, M.D., M.S., is a well-funded, highly productive expert in sleep-disordered breathing who has more than 20 years of experience in sleep medicine and a long history of mentoring K awardees and junior faculty. Two carefully selected co-mentors, physiologist Louise O'Brien, Ph.D., M.S., and psychiatrist Juan Lopez, M.D., will provide focused training in perinatal sleep and neuroendocrinology, respectively. The proposal is also bolstered by a tailored consultant team. Research. The goal of this application is to understand the contribution of sleep-disordered breathing (SDB) to one of the most common and debilitating adverse pregnancy outcomes, perinatal depression. It is our hypothesis that positive airway pressure (PAP) treatment of SDB in pregnant women with comorbid depression will result in significant reductions in depressive symptoms and improved neuroendocrine functioning. This proposal will leverage our unique expertise with PAP for SDB in pregnancy and adverse pregnancy outcomes, including perinatal depression. The rationale is that a better understanding of the role of SDB and the impact of PAP treatment on perinatal depression will allow for development of targeted and critically-timed interventions which may have long-term benefits for both women and their offspring. We plan to test our hypothesis by randomly assigning 50 women with SDB to receive either positive airway pressure treatment (PAP group) or treatment as usual in obstetrics (non-PAP group). The following specific aims will be tested: AIM 1: Determine the impact of PAP treatment on prenatal depressive symptoms in pregnant women with SDB and depression. AIM 2: Identify the contribution of PAP treatment on the HPA axis in pregnant women with SDB and depression. An exploratory aim is to assess associations between PAP treatment and adverse pregnancy and neonatal outcomes in women with depression and SDB. With respect to expected outcomes, the proposed work will demonstrate that SDB during pregnancy plays a role in perinatal depression, and that treatment of SDB during pregnancy will ameliorate depressive symptoms and HPA axis functioning, and improve pregnancy and neonatal outcomes. Such findings are expected to have an important positive impact, because they will further our understanding of the role of SDB in adverse pregnancy outcomes and reduce the burden of the most common and debilitating adverse outcomes experienced by pregnant women. This will have long term health benefits for both women and their offspring. Summary. The proposed K23 will provide key additional training for a promising early investigator to become a clinical researcher in perinatal sleep medicine. The project will also address important, testable questions about the relationships between a common sleep disorder and the debilitating adverse pregnancy outcome of perinatal depression, a major public health concern.


Major depressive disorder (MDD) is a debilitating disorder of altered mood regulation that is precipitated by chronic stress, modulated by serotonin and of unknown molecular pathophysiology. Four prominent risk factors have been consistently reported to influence rates of vulnerability to develop depression: sex, genetic make-up, prior MDD episodes, and early life stress;however no single animal model has comprehensively combined these vulnerability factors. Using the unpredictable chronic mild stress (UCMS) protocol and behavioral assessments of anxiety/depression-like behaviors in serotonin transporter (SERT) mutant and control mice, I have modeled the interactions of sex, SERT genetic make-up, and stress in eliciting depressive-like behaviors, including the increased vulnerability to clinical depression observed in human female subjects and the risk that is conferred by low SERT levels. Furthermore, gene microarray studies performed by our laboratory on the amygdala of postmortem human MDD subjects and UCMS-treated mice have identified ~40 genes whose changes are specific to human MDD and rodent UCMS, and reversed by antidepressant treatment in rodents, thus representing a critical pool of genes differentially expressed according to altered mood. Therefore, in addition to modeling aspects of human susceptibility to develop depression (sex and SERT genetic make-up), the rodent UCMS paradigm induces molecular changes that are predictive of ?depressive states? across species. Using a comprehensive UCMS-based experimental design in the mouse, I will first confirm the role of sex and SERT as risk factors to UCMS-induced altered mood regulation and then extend my investigation to two additional factors: disease recurrence and early life stress (Aim 1). Using samples generated in Aim 1, I will then begin characterizing neurobiological phenotypes that underlie the UCMS-evoked depressive-like state (Aim 2), including neuroendocrine changes and quantitative assessment of the amygdala gene expression signature. I hypothesize that while risk factors may differentially increase the susceptibility for developing MDD, the ultimate state of altered mood regulation affects a common set of biological and/or molecular disturbances. Results from these studies will begin characterizing changes in these putative common molecular and neuroendocrine disturbances, will reveal the relative contribution of these susceptibility factors to this common phenotype, and will form the basis of future causative studies aimed at investigating the cellular and molecular basis of MDD.


AIDS stigma, or negative meanings surrounding people living with HIV/AIDS, has been fostered by myths and false notions surrounding the epidemic (Herek & Glunt, 1988, p. 886). AIDS stigma has multiple negative psychosocial consequences including anxiety, depression, disruption of family dynamics, physical and emotional violence, loss of social support and the deterioration of productive relations with health-related professionals (Chesney & Smith, 1999; Kalichman, 1998b; Nord, 1997). Given the growing number of HIV cases among the Latino community in general, and Puerto Ricans in specific, it is necessary to address the eradication of AIDS stigma, particularly among professionals who interact with HIV+ individuals such as those in health related fields (Nord, 1997). AIDS stigma has been documented among health services providers such as doctors, nurses, mental health professionals, social workers and other caregivers (Acuff, et al., 1999; Gordon, et al., 1993; Herek & Glunt, 1988; Rizzo, 2002; Silverman, 1993; Stevenson & Storhm Kitchener, 2000; Trezza, 1994; Ventura, 1999). These stigmatizing attitudes towards HIV+ people are unfortunate, especially when research has reported those who feel stigmatized by health professionals face problems accessing optimal health services (Link & Phelan, 2001; Valdiserri, 2002; Weiss & Ramakrishna, 2001). As recently as 2002 stigmatizing attitudes among health professionals in Puerto Rico have been documented by people living with HIV/AIDS. Findings suggest the need to address AIDS stigma through effective interventions that incorporate other stigmas surrounding the epidemic such as negative attitudes towards sexual orientation, drug users, and women. In order to do so, stigma combinations surrounding HIV/AIDS among health professionals must be explored. The main objectives of this research application are to: 1) to explore stigmatizing attitudes towards people living with HIV/AIDS among health professionals in Puerto Rico, 2) to explore the interaction of other stigmatizing attitudes towards gender, sexual orientation, and drug use with AIDS stigma in the same population, and 3) to refine and pilot test an intervention module for stigma reduction geared towards these professionals. These objectives will be achieved through the implementation of a sequential mixed method study using qualitative in-depth interviews and a quantitative self-administered questionnaire addressing AIDS stigma.


Osteoporosis, a disease characterized by the systemic loss of bone mass and strength resulting in fragility fractures, is rapidly poised to become a major public health threat in the United States. Osteoporosis results from imbalances in bone remodeling, a process characterized by osteoblast-mediated bone synthesis and osteoclast-mediated bone resorption. Because osteoporosis is fairly asymptomatic, and is often detected only after the patient has sustained significant bone erosion, therapies aimed at restoring the eroded bone are equally important as those that target bone resorption. However, unlike the highly efficient repertoire of anti- resorptive drugs that form the mainstay of the current anti-osteoporosis therapy, drugs that stimulate bone formation remain largely underdeveloped. Hence there is a critical need for novel therapeutic targets that will stimulate osteoblast-mediated bone accrual together with the inhibition of osteoclastic bone resorption. Our preliminary studies identify Ca2+/calmodulin (CaM)-dependent protein kinase kinase 2 (CaMKK2) as one such target as its inhibition positively impacts anabolic pathways and negatively impacts catabolic pathways of bone remodeling. Mice null for CaMKK2 possess enhanced trabecular bone mass in their long bones, along with significantly higher numbers of osteoblasts and fewer multinuclear osteoclasts. Moreover, its inhibition offers protection from ovariectomy-induced osteoporosis in mice. The proposed studies will enable us to define the precise mechanism by which CaMKK2 regulates osteoblast and osteoclast differentiation and devise potential strategies of its inhibition in the treatment of osteoporosis. Development of CaMKK2 inhibition as a new generation therapeutic target that promotes robust bone mass accrual while inhibiting resorption will represent a major breakthrough in anti-osteoporosis treatment.


A panel of hybrids of HeLa and a normal human fibroblast differing in tumorigenicity in nude mice was screened for RNA levels specific for 21 oncogenes and for human papillomavirus 18 and for papillomavirus were detected. The levels of three of these oncogenes. Transcripts for 15 of these oncogenes were found to correlate with tumorigenicity. These were fos and ets transcripts, which were present at a higher level in tumorigenic hybrids than in non-tumorigenic hybrids and myb transcripts, which were higher in the non-tumorigenic hybrids. Fos protein levels correlated with fos transcript levels. A non-tumorigenic hybrid, when infected with murine sarcoma virus, did not show alteration in the levels of fos, ets or myb transcripts. A panel of human SV80 cells transfected for the proviral mos gene has been generated. These transfectants differ in tumorigenic potential in nude mice. Explants of the tumorigenic members of this panel do not show any alteration in the size or amount of immunoprecipitable SV40 T antigen consistent with an immunological selection against T antigen expression during tumor formation. RNA has been prepared from the members of the panel for an examination of the levels of mos-specific transcripts. An activated N-ras gene cloned previously from human gastric adenocarcinoma material has been characterized using DNA sequencing and transfection assays of in vitro ligated DNA fragments.


The overall objective of this project is to understand the role of E2F transcription factor in regulation of cell proliferation and how a loss of this regulation can lead to oncogenesis. Indications that the cellular transcription factor E2F plays a role in cell proliferation control was obtained from recent analyses which demonstrated a physical interaction between the tumor suppressor protein Rb and E2F. Further, the loss of Rb function is closely associated with a loss of the Rb-E2F interaction, suggesting that E2F is a functionally relevant target for Rb. Additional proteins involved in cell cycle regulation like p 107, cyclins A and E, and the kinase cdk2 are also found to be associated with E2F, indicating that E2F could be playing a major role in regulating cell proliferation. This project aims at studying the role of E2F in tumorigenesis, differentiation and cell cycle regulation. It will be assessed whether there is correlation between the loss of the Rb-E2F interaction and the generation of a wide variety of human tumors. In addition, the possibility that a mutation of E2F can lead to oncogenesis will be examined. To further understand the mechanism of E2F action, additional proteins that interact with E2F will be identified and cloned. Experiments will also be conducted on the role of E2F and associated proteins in the differentiation process of cells. Finally, attempts will be made to identify the cellular promoters that are targeted by E2F an E2F associated proteins.


Tolerance to nicotine develops with chronic smoking and may be associated with the onset of tobacco dependence. It is unclear how quickly this tolerance dissipates after terminating exposure to nicotine. We propose to longitudinally assess responses to measure doses of nicotine by nasal spray in 30 male and female smokers as they smoke ad lib for two days (baseline), quit for six days, and then resume smoking for two days on the GCRC. We hypothesize that responses to nicotine will become larger over the six days of cessation, showing dissipation of tolerance and then be smaller during the final two days of ad lib smoking, showing reinstatement of tolerance with return to smoking.


PROJECT SUMMARY/ABSTRACT ! There is a looming social and economic liability with a growing elderly population. Annually, a staggering 2.8 million elderly are treated in emergency departments for fall injuries. Falls are the leading cause of traumatic brain injury and the primary cause of accidental death in adults over 65 years. Impaired balance, particularly in the lateral direction, is one of the most important risk factors for falls. Unfortunately, common clinical tests of balance are not sensitive to impairments. This lack of sensitivity makes it difficult to monitor changes in balance control that may increase a person's fall risk (e.g. following a change in medication). Therefore, there is a critical need for more sensitive tests to assess balance. We have shown that delays in neuromuscular control result in instability. This work along with the fact that elderly have slower postural adjustments prompted our research group to develop a test to assess speed of balance control. We have shown that speed of balance control can be reliably assessed, is sensitive to subtle impairments in balance not found with common clinical tests, and has ecological validity as demonstrated by predicting performance in common daily activities known to be predictive of falls. The product of this SBIR is a low-cost, easy-to-use system to assess speed of balance control and predict fall risk. In this supplemental award, three members of the research team will attend the National I-CorpsTM program and perform 100 or more customer discovery interviews. By conducting these interviews, we will increase the breadth and depth of knowledge of the various healthcare ecosystems involved in reducing falls in the elderly and identify pain points and barriers for clinical adoption of our balance assessment technology. This information will be important for learning who and what will drive or resist technology adoption. !


A systematic investigation to evaluate the effect of ionizing radiation upon liver microcirculation is proposed. By using objective methodology radiation induced changes in oxygen transport to tissue, blood cell aggregation, liver function liver scans, histology and ultrastructure will be determined to ascertain acute as well as late effects. Reported results of a pilot experiment show the feasibility of this approach. After proper quantitation of the inflicted insult, prevention or reversal will be attempted by systmatic testng of peripheral circulation flow improvers, such as low molecular weight Dextran, and drugs known to prevent intravascular red cell and platelet aggregation or adhesiveness such as anti-adhesive drugs and aspirin. Successful completion of these experiments will immediately provide a method to allow radiation treatment of liver cancer, metastatic or primary, a disease today lacking a cure except for chemotherapy palliation.


This study is an extension of the original placebo-controlled trial to determine the safety and efficacy of Copolymer I in patients with relapsing-remitting multiple sclerosis. The controlled phase 3 trial showed that treatment with copolymer 1 was effective in reducing the clinical relapses and in slowing accumulated neurological disability compared to placebo treated patients. Most patients completing the phase 3 placebo controlled study elected to enroll in this open-labeled extension study of copolymer 1 for determination of long term safety and apparent persistence of efficacy. The study will be extended for a minimum of two additional years with the evaluation of all enrolled patients with annual quantitative magnetic resonance imaging, with centralized image analysis conducted at the University of Texas-Houston, Health Science Center.


This proposal focuses on investigating the mechanism of synaptic transmission at the inner hair cell (IHC) afferent synapse in the mammalian cochlea. This most peripheral chemical synapse in the auditory pathway serves as an interpreter translating the hair cell receptor potential into a train of excitatory postsynaptic potentials that activate action potentials in the afferent auditory nerve fibers. The specific features of the IHC afferent synapse critically determine how a sound signal is coded in the inner ear and transmitted to the brain via auditory nerve fiber activity. Our aim is to characterize pre- and postsynaptic mechanism that shape the postsynaptic activity in IHC afferent fibers. We will record postsynaptic activity using the whole cell patch clamp method. The recording sites will be the afferent fiber terminal directly at the inner hair cell synapse and the afferent fiber soma in the spiral ganglion in excised preparations of rat and mouse cochleae. To exactly describe the relationship between presynaptic stimulus and postsynaptic activity, we will make double recordings from IHCs and afferent fiber terminals. To affect transmitter release and define the quantal size, we will record excitatory postsynaptic currents and vary experimental conditions; i.e. changing the temperature or ionic environment and using release-affecting toxins. To understand postsynaptic determinants of synaptic transmission, we will provide a basic description of the waveform of the excitatory postsynaptic potential and its relation to the activation of action potentials. Using pharmacological tools, we will try to identify individual components that shape the postsynaptic potential waveform starting with a focus on different classes of potassium channels. We will also study the role of efferent inputs onto afferent fiber activity. Most recordings will be performed in the already established preparation using postnatal cochleae. In parallel we will extend our recordings to mature cochleae, as our ultimate aim is to understand both development and adult function of the IHC afferent synapse. These studies will further our knowledge of basic mechanisms of peripheral hearing and therefore also will lead to a better understanding of abnormal hearing and deafness.


The Office of Education delivers workshops, programs, and individualized opportunities to a population averaging 300 trainees, including postdoctoral, visiting, and research fellows; clinical fellows and medical students; graduate students; and postbaccalaureate fellows, as well as summer trainees. The activities include, typically: public speaking workshops, job interviewing, writing and editorial services, grantsmanship workshops, career presentations and counseling, teaching opportunities through the NICHD Becoming an Effective Scientist course for postbacs and skills workshops and a training program with the University of Maryland, and management programs. An annual retreat for fellows and graduate students is held each spring to address scientific developments and careers, which includes two keynote addresses, fellow presentations, NICHD alumni leading career round tables, and a poster presentation by each attendee. The program is developed and run by a fellow-student steering committee. Among the office's accomplishments from the past year: Dr. Yvette Pittman continues in the Office of Education as Associate Director. Our TmT (Three-minute Talks) competition, now in its third year, is held in conjunction with NIDCR and NHGRI. Dr. Alex Szatmary in the laboratory of Dr. Ralph Nossal received the third-place award. The office developed an online Annual Progress Review for fellows, launched in 2016, to track scientific and career development and progress; APRs will be provided for BSC site visits of our investigators. The database of NICHD alumni from 2008 to the present continues to be updated, leading to an expansion of our Linked In alumni group. In June 2016, the Division of Intramural Research gave its ninth Mentor of the Year awards to Julian Lui, Ph.D., in the investigator category; and Shlomo Krispin, Ph.D., fellow. Twenty FARE awards were made for the 2017 competition. A Fellows Intramural Grants Supplement (FIGS) continues to recognize and stimulate grant applications among fellows, and the Fellows Recruitment Incentive Award (FRIA) continues to support investigators who recruit postdocs from populations traditionally underrepresented in science. The NICHD Scholars program, in its sixth year, had one PhD graduate and one postbac advance to medical school; of three new postbac fellows, two are continuing for a second year, one is pursuing public health training,and two new postbacs are joining this group. The Scholars program focuses on developing talent and supporting trainees' academic and career progression. The NICHD Connection monthly newsletter published its 75th issue in August 2016 and reaches all members of the intramural division and our alumni.


The principal objective of the research is to develop an ultrasonic dosimetric model or models from which the ultrasonic energy being absorbed by or interacting with tissue can be determined. The emphasis will be to define physical quantities which properly reflect an ultrasonic energy deposition in situ based upon knowledge of the free field distribution and ultrasonic propagation properties of the material. The second objective is to develop a dosimetric format by which patient exposure and particular organ and other body site exposure to ultrasonic energy is determinable. The anticipated format would be in terms of a quantitative parameter, or parameters, and would be useful in any subsequent retrospective or propective epidemiology study. The basic procedure will be to account for all of the ultrasonic energy when a specimen is irradiated. This will be accomplished by measuring the detailed ultrasonic field distribution patterns in the free field, in the field with the specimen in place and within the specimen. The measurement procedures include three absolute techniques and one relative technique. The specimens include phantoms of known acoustical properties and the biological tissues, testis, liver and kidney.