David Smith - US grants

The research will describe the neuroanatomical and neurochemical bases of ketamine's (Ketalar; Ketaject) analgesic effect. It will be accomplished by an evaluation of the presence and function of opiate receptor sub-types in neuroanatomical loci (i.e., sites known to be loci of morphine's analgesic action); a description of opiate receptor sub-types with which ketamine interacts; and a determination of the consequences of an interaction of ketamine at these receptors. The research is divided into five scientific approaches. Each is designed to provide component information important to the project as a whole and/or to test the same proposal under different conditions. The methods are: 1) a pharmacological comparison of the analgesic action in rats of various sub-classes of opiate drugs presumed to be specific for sub-types of opiate receptors, and an evaluation of the participation of monoaminergic and opiate neuronal processes in these analgesias; 2) an in vitro comparison of the receptor binding characteristics of ketamine and other opiate drugs, specific for different receptors, using opiate binding assays; 3) an in vivo evaluation of the ability of ketamine to interact with opiate receptors that may mediate its special pharmacological actions; 4) local administration of ketamine and opiate drugs, specific for different opiate receptors, into central nervous system loci (i.e., those sites responsible for morphine analgesia) and an analysis of the neurochemical and neuroanatomical bases for the resulting analgesia and 5)*an in vitro and in vivo analysis of the participation of opiate or non-opiate mechanism(s) in the spinal analgesic action of ketamine, a site of action of established differences between ketamine and morphine. The study should define ketamine's opiate receptor preference and action (agonistic-antagonistic) on various sub-types of receptors found in areas of the central nervous system associated with morphine's analgesic action. The possibility that ketamine uses neuronal circuitry activated by morphine is being tested. Ketamine may be found to be dissimilar to morphine and share components of analgesic mechanisms associated with other classes of opiate drugs. Differences in the analgesic mechanisms associated with various types of analgesic drugs will begin to be defined which is consistent with our long term goal to determine pathways and neurotransmitters of analgesia.*

Although there has been considerable progress in understanding the central neural processing of taste information and the receptor and transduction mechanisms for taste stimuli, including several recent advances from molecular biology, the link between different levels of analysis remains elusive. Studies conducted during the previous funding period have provided a much better understanding of the relationship between receptor mechanisms, central neural responses, and behavior elicited by sodium and nonsodium salts and acids. However, there is little information about such relationships for stimuli that humans classify as sweet or bitter. The studies proposed in this application seek to link taste receptor cell (TRC) activity to the responses of central gustatory neurons and behavior in rats. These experiments use a newly developed method for recording from TRCs in intact gustatory epithelia to examine the distribution of sensitivities to sweet and bitter stimuli and to relate these data to responses to these stimuli in rat brainstem neurons and to stimulus relationships established in rat behavioral experiments. These studies address current questions in gustatory neurobiology and test several underlying hypotheses about the coding of information for stimuli described as sweet or bitter by humans. The first Specific Aim, which will continue a collaborative effort with Dr. Tim Gilbertson, utilizes a newly developed method for recording from TRCs in intact fungiform and palatal epithelia of the rat to investigate the distribution of sensitivities to bitter and sweet stimuli in these cells. These studies test the hypothesis that receptor cells vary in their sensitivities to stimuli within the bitter and sweet categories. In addition, transduction pathways for diverse bitter substances will be investigated in this preparation using pharmacological blockers. The second Specific Aim involves single-neuron recording from the rat NST to investigate the central neural coding of sweet and bitter stimuli. These studies test the hypothesis that the patterns of central neural activity generated via the VIIth nerve can be predicted from the distributions of receptor activity demonstrated in Aim 1. The third Specific Aim employs behavioral methods to generate psychophysical functions in rats to arrays of sweet and bitter stimuli. These functions will be used to derive similarity indices among these stimuli to test the hypothesis that behavioral similarity can be predicted from receptor and brainstem responses recorded in Aims 1 and 2. The overarching goal of these proposed experiments is to determine, in a single species and with the same stimuli, how the activity of individual TRCs contributes to the central neural representation of behaviorally categorized stimuli.

The objectives are to continue development of magnetic resonance spectroscopy (MRS) as a "tool" for exploring bioenergetic events in the brain in vivo. The present work proposes to explore two hypotheses: 1. That the severity of an hypoxemic insult in the adult brain can be predicted by MRS determination of phosphocreatine (PCr) concentration. 2. That brain work and brain bioenergetics are related by a Michaelis-Menten hyperbola and that metabolic instability can be predicted from MRS measurements. In pursuing these two hypotheses, the changes obtained with MRS will be related to measures of oxygen availability, brain redox state, cerebral metabolism of oxygen and glucose, cerebral production of lactate, brain intracellular pH, cerebral blood flow, neurophysiologic function (electroencephalography and somatosensory evoked potentials), and histopathology (by both light and electron microscopy). Finally, the findings with MRS will be compared to measurements of brain metabolites by standard biochemical techniques to determine the sensitivity and reproducibility of MRS measurements compared to traditional approaches. The experiments will be done on anesthetized dogs, intubated, mechanically ventilated and prepared for invasive monitoring of vital signs. Two different types of experiments will be done: In the first, the dog will be exposed to a period of "stabilized hypoxemia" in which the degree of metabolic stress is continuously adjusted according to "real time" determinations of PCr by MRS. At various times oxygen will be restored and the rate of recovery or the development of metabolic instability will be determined. In the second type of experiment, brain work will be increased by administration of a short acting epileptogenic gas. The ability to finely titrate the dose of this agent should allow graded increases in cerebral metabolic rate that will allow determination of the relationship of brain work to brain energy. These experiments have direct relevance to medicine. Many catastrophic brain events involve changes in brain energy production. MRS is the only technique with the potential of determining these changes in man, since it can measure brain energy non-invasively and non-destructively. Basic studies of MRS in animals are required to understand the meaning of the measurements when they are obtained from humans. In addition, the studies will add to the fundamental knowledge concerning the production and utilization of energy in the brain.

Although the gustatory components of food almost always occur as complex mixtures, most research in taste, particularly the neurophysiological work, has employed simple stimuli, representing the four basic taste qualities of sweet, salty, sour and bitter. Mixtures of substances with different tastes or the interaction of similar-tasting stimuli has received relatively little attention by electrophysiologists. The studies described in this proposal are designed to investigate the neurophysiology of homogeneous (same quality) and heterogeneous (different quality) mixtures of taste stimuli. The responses of single neurons in the parabrachial nuclei (PbN) of the hamster will be recorded extracellularly. Stimuli will be concentration series of sucrose NaCl, citric acid, and quinine hydrochloride (QHCl) and their mixtures (heterogeneous) and sucrose, fructose, xylitol, and Na-saccharin and their mixtures (homogeneous). Three heterogeneous mixtures will be studied: sucrose - NaCl, sucrose - QHCl, and NaCl - citric acid. Homogeneous mixture experiments will include sucrose - fructose, sucrose - Na-saccharin, and sucrose - xylitol mixtures. These stimulus pairs were selected on the basis of known mixture interactions in human psychophysical studies or in electrophysiological studies on the chorda tympani nerve. For a given cell, concentration series for each of a pair of stimuli will be presented, followed by several mixture series, in which different concentrations of one of the components will be added to the concentration series of the other. Thus, several concentrations of a pair of stimuli will be combined factorially so that the responses of individual PbN neurons can be assessed relative to predictions made from several models of stimulus-receptor interactions, which have been used in both electrophysiological and psychophysical studies of taste mixtures. The quantitative nature of the mixture interaction for each pair of stimuli will be compared across neuron types in the PbN, defined by their response profiles to stimuli representing the four taste qualities. Deviations from these models or differences in the response to a given mixture pair between neuron types will suggest a role for neural interactions in the mixtures. The mutual suppression between heterogeneous mixtures, which has been noted in human psychophysical studies, will be correlated with the occurrence of inhibitory responses, which are more frequently seen in these third-order gustatory neurons than in more peripheral cells. Interactions among stimuli showing mixture suppression or synergism in the PbN will be studied in single fibers of the chorda tympani nerve to further assess the contribution of synaptic processing to mixture phenomena.

This project will develop ethical guidance for presymptomatic testing for autosomal dominant, late onset diseases for which genetic probes are available (familial Alzheimer disease, amyotrophic lateral sclerosis, Huntington disease, myotonic dystrophy, neurofibromatosis, adult polycystic kidney disease and retinitis pigmentosa). Our six-member working group offers expertise in genetic counseling, clinical medical ethics, genetic research, law, ethics and communication. We will collect and analyze case studies, beginning with Indiana University's extensive experience with Huntington disease (HD). Experience with HD research and presymptomatic testing provides a unique and informative historical model. Co-PI Kimberly A. Quaid will visit selected testing centers, and we will solicit supplementary cases from major centers that offer testing for diseases of interest. We will define the full range of ethical problems presented by presymptomatic testing for these disorders, and we will explore the salient questions from the varied perspectives represented by the working group, considering ethical principles (autonomy, beneficence, justice); clinical practicality; administrative feasibility; changing research findings; religious beliefs and institutions; and political, cultural and economic contexts. Consultants P. Michael Conneally, Robert Burt, Albert Jonsen and Thomas Murray will read drafts of our cases, analyses and guidelines and provide regular feedback on our work throughout the project. In addition, we will solicit input from genetic counselors and others experienced with presymptomatic testing, and we will invite patients and families at risk to review our materials in progress to assess their sensitivity, adequacy and feasibility. Our final product, a book published by Indiana University Press, will include guidelines for presymptomatic testing of autosomal dominant, late onset disorders; annotated cases; and the description of a method for resolving ethical issues our guidelines do not address directly.

This project utilizes behavioral and physiological techniques in the same animals to determine the functional role of olivocochlear bundle (OCB) efferents in auditory perception. Subjects will be monkeys (Macaca fascicularis). These will tie trained to respond on perceptual tasks using operant conditioning techniques applied in a standard yes-no adaptive tracking procedure. Perceptual processes to be examined are (1) monaural thresholds in quiet and under conditions of broadband noise masking, (2) frequency selectivity, (3) dynamic range and growth of loudness, and (4) central masking. These tasks were selected for study because they allow hypotheses of OCB function previously described in the auditory research literature to be evaluated. Baseline data for normal performance on one or more perceptual tasks will be collected for each animal. The animal will then undergo either a unilateral vestibular neurectomy (to remove the entire OCB projection to one ear) or near-midline section at the floor of the fourth ventricle (to eliminate only the crossed OCB projections). Following a recovery period, animals will be re-tested on the same tasks as earlier to determine the perceptual consequences of OCB loss. After behavioral testing has been completed, each animal will become the subject of an acute electrophysiological experiment to record single auditory nerve fiber responses, using the same stimuli and data collection paradigms that were used behaviorally. At the conclusion of the physiological recordings, the operated and unoperated cochleas of each animal will be injected with horseradish peroxidase and nuclear yellow, respectively, and light and electron microscopic techniques will be used to verify OCB transection (for the operated ear) and determine the number and condition of surviving OCB cells (for the unoperated ear).

9451165 Bart The field of geological education has changed dramatically in the past 2O years. Part of this progress is reflected in the incorporation of environmental and hydrogeologic courses in the curriculum. Undergraduates engaged in geologic study throughout our nation are moving into these endeavors because of the strong employment possibilities created by a growing industry. Until recently, environmental and hydrologic study were not part of the curriculum at La Salle University. The Department of Geology is hoping to correct this deficiency in our program by purchasing the necessary equipment to add hands-on environmental and hydrogeologic experiences to our curriculum. This equipment will be used by freshmen, sophomore, junior, and senior majors, in earth science teacher preparation, and freshmen introductory classes, and will serve as an important part of our curriculum in: Introductory Geology, Environmental Geology, Hydrogeology, Sedimentology, Environmental Geology, and Student Research. With this laboratory equipment our students will be complementing their intensive use of standard geologic investigation equipment and will be capable of investigating the earth in ways never before possible in our program. Subsurface interface radar will be used for conducting studies such as bedrock depths, soil profiles, water table information, fracture and fault mapping. Water studies will include water analysis for characterization of surface and subsurface quality. The gas vapor soil probe and organic vapor meter will be used for investigations of possible hydrocarbon contamination of soils. The radon detector will give students the chance to evaluate radon levels of soils throughout our region. If supported, this project will create a new and exciting dimension in geologic investigation at La Salle; and the students we serve will be better prepared for the challenges of graduate studies, teaching, employment in environmental sciences, and research.

Liarozole is a benzimidazole compound that is currently under development for the treatment of malignant disease. Liarozole has been shown to raise endogenous levels of retinoic acid in animal models, probably by binding to the cytochrome P450 moiety of the 4-hydroxylase responsible for retinoic acid catabolism. This study will assess the effect of liarozole on the P450 system and markers of retinoid metabolism and effect in humans.

This project is concerned with the behavioral analysis of the biophysical mechanisms underlying cochlear implant function. Behaviorally trained cats will be deafened unilaterally and implanted with one of two multi- contact intracochlear electrode arrays. Two classes of experiments are proposed. The first is to relate psychophysical performance to anatomic findings and several neurophysiological measures (EABRs, intracochlear- evoked potentials, and single-fiber discharge patterns) in response to the same stimuli, all in the same animals. Behavioral tasks include measures of absolute threshold, growth of loudness, dynamic range and channel interactions. In the second class, the goal is to characterize the effects of minute manipulations of intracochlear current fields on psychophysical performance. Using a dense, multicontact linear-array electrode, absolute thresholds for single and paired pulsatile stimuli will be measured as a pulse phase polarity and electrode position. Intracochlear-evoked potentials, EABR and electrical field mapping studies will be undertaken to characterize the growth in electrical field and activation, as a function of stimulus intensity, for the same stimuli as were used behaviorally. At the conclusion of the psychophysical and electrophysiological testing, animals from both classes of experiments will be sacrificed, and their cochleae harvested for histological analyses to determine the number and spatial position of surviving peripheral prncesses and spiral gang lion cells, relative to the electrode contacts. Data from both classes of experiments, combined with predictions of electrical fields from a finite element model, will form inputs into biophysically-based, neural behavioral models. The models will be evaluated in an attempt to account for behavioral performance on the basis of electrical field configuration, the resulting patterns of neural activity, and the anatomic condition of the spiral ganglion in the tested animal. In addition, the feasibility of conducting fine focal stimulation within the cochlea, both within and across a critical band length, will be evaluated in terms of measures of channel interactions and behavioral performance.

Calcitriol is the active form of vitamin D. Calcitriol plays a role in normal and neoplastic cell growth and can modulate growth through specific receptors resulting in differentiation and growth inhibition. This study will evaluate the ability to administer calcitriol via a parenteral route in cancer patients alone and in combination with prednisone which has been shown to reduce hypercalcemia and potentiate the antitumor effects.

DESCRIPTION (Adapted from applicant's abstract): The overall goal of the proposed research is to identify and characterize the neurophysiological substrates of certain context-appropriate behaviors. Since the learning context plays an important role in normal learning and memory, developing an understanding of the neuronal mechanisms that underlie context-specific behavior is an important step towards understanding how the brain processes information for future use. More specifically, this project seeks to test the hypothesis that the hippocampus signals contextual information to the cingulothalamic system to produce special patterns of neuronal activity that mediate context appropriate associations and behaviors in a discrimination learning paradigm. This will be accomplished by recording the activity of neurons in cingulothalamic circuitry and the behavior of control rabbits and rabbits with temporary and permanent lesions of the hippocampus while they perform two different discriminative learning tasks in different environments.

This pilot study will assess the safety/tolerance and length of time that plasma levels of unchanged paclitaxel exceed 0.07 uM after oral administration of paclitaxel. 18 adult patients with stable metastatic prostate, breast or lung cancer will be enrolled. After an overnight fast, the patients will receive 5 mg/kg cyclosporin oral solution dose (Sandimmune) 1 hour prior to and concomitantly with oral paclitaxel at one of three planned doses.

Prostate Cancer is the most common malignancy and the second leading cause of cancer death in males in the United States (1). Despite advances in localized therapy of prostate cancer, a significant fraction of men develop metastatic disease at some point in the course of the illness. We have previously studied treatment approaches for this disease which focus on the nuclear matrix and the DNA replication complex. This protocol will continue those studies using the topoisomerase I inhibitor irinotecan (CPT-11) in combination with estramustine.

DESCRIPTION: (Adapted from the Investigator's Abstract) The ultimate goal of the proposed studies is characterization of the role of the medial efferent system in human perception of sound. It combines psychophysical and physiological measurements in the same subject, from both humans and monkeys. The PI proposes the development of psychophysical tasks that reflect known efferent processes. The characteristics of the distortion product otoacoustic emission (DPOAE) will be used to define the stimulus conditions for the specific aims. Based on psychophysical and anatomical studies of the two species, use of identical stimulus conditions allows direct comparison between the two species. The first study (Aim I) is designed to determine the relationship of "central masking" and efferent reduction of the DPOAE, both effects produced by contralateral acoustic stimulation. Psychophysical suppression will be measured parametrically by target frequency, contralateral noise bandwidth, intensity and frequency. In human subjects, the contralateral efferent effect on the DPOAE will determine the efferent involvement. Similar experiments, using identical stimulus conditions, will be performed with the monkey as the subject. The efferent involvement in perception, measured from the human, will be inferred from monkeys with discrete lesions. The experiments performed in Aim I will aid in the specification of stimulus conditions of subsequent Aims. In Aim II the PI proposes to characterize the role of the medial olivocochlear (MOC) in possible improvement in the perception of signals in noise. These studies are intended to determine the conditions in which enhanced perceptual acuity occurs. Aim III focuses on the role of the MOC in psychophysical adaptation. Recent physiological experiments suggest that some aspects of adaptation are related to MOC function. The DPOAE in response to sustained stimulation shows a rapid adaptation that is eliminated when the uncrossed efferent system is cut. The PI proposes to test, in parallel, psychophysical and physiological adaptation in humans and monkeys. The time course of the adaptation for each response measure will be compared. The involvement of the MOC in these two response measures will be evaluated by MOC lesions in the monkeys. The goal of Aim IV is to determine the role of the MOC tracts in auditory "selective" attention. Psychophysical and physiological evidence suggests that a control mechanism exists that can suppress extraneous or unattended signals. Parallel psychophysical and physiological studies in humans and monkeys are proposed to replicate the previous studies. The role of selective attention will be addressed by discrete MOC lesions and monitoring performance changes.

taste buds; phenotype; taste; cell type; voltage /patch clamp; dyes; genetically modified animals; laboratory mouse;

The medial olivocochlear (MOC) efferent system is known to exert significant effects on cochlear physiological activity, yet little is known about the function of the efferents in auditory perception or the behavioral response of hearing. The proposed functions of the MOC, relevant to the present work, include: 1) to unmask or decompress an auditory nerve fiber's I/O function by suppression of the neural representation of concurrent noise; 2) to increase the salience of transient signal onset by mediating the process of neural adaptation, which acts to decrease or suppress the representation of long-duration stimulation; and 3) to provide a peripheral, neural mechanism that underlies the process of "selective" auditory attention, by which the neural encoding of "unattended" stimuli are suppressed. This application proposes a unique experimental approach to characterizing the role of the efferent tracts in auditory perception in humans, combining psychophysical and physiological (DPOAE) measures in the same subjects. We propose to develop psychophysical tasks as analogs of known efferent physiological processes, focusing on psychophysical measures related to the above, purported efferent processes. Specific psychophysical tests will employ both human and non-human primate subjects as listeners, in parallel, under identical stimulus conditions. The DPOAE measures will serve to aid in specification of the precise stimulus conditions to be tested, being optimized for specific aims. It is assumed, based on previous psychophysical and anatomical studies, that the humans and monkeys will yield comparable data under similar stimulus conditions. The ultimate goal of this research program is characterization of the contribution of the medial efferent tracts to auditory perception in humans. Parallel testing of human and non-human subjects permits unequivocal determination of the contribution of the efferents to the different perceptual processes studied, in both monkeys and humans, through lesion studies in the monkey subjects. Without this parallel testing paradigm (and the ability to lesion tracts in the non-human primate subjects) the link between human perception and the underlying neural mechanism would not be direction obtainable.

DESCRIPTION (Provided by applicant): Angiogenesis is a critical factor in the progression of malignant disease. Malignant cells overexpress several factors that stimulate the process of vascularization including vascular endothelial growth factor (VEGF) and basic fibroblast growth factor (bFGF). Copper plays a critical role in the process of angiogenesis, acting as a cofactor for the function of several of the key mediators of angiogenesis. The copper-depleting agent tetrathiomolybdate (TM) has been shown to modulate the process of angiogenesis by inhibiting the production of these proangiogenic agents. Both benign and malignant prostate cells express VEGF and bFGF. Recent evidence shows that these molecules are differentially expressed with disease progression in prostate cancer. Urinary VEGF levels correlate with survival in patients with advanced disease indicating the importance of angiogenesis in the progression of this disease. This phase II clinical trial will evaluate the activity of an antiangiogenesis strategy using tetrathiomolybdate to deplete copper in patients with prostate cancer. Assessment of the efficacy of therapy in prostate cancer is difficult due to the predominance of bone metastases in this disease. Prostate specific antigen (PSA) has been shown to be a marker of response following hormonal and cytotoxic therapy for prostate cancer. In a laboratory model, PSA has been shown to inhibit angiogenesis, complicating its use as a response indicator to monitor the effect of therapies aimed at inhibiting angiogenesis. This study will evaluate PSA as an indicator of response to antiangiogenic therapy by correlating levels with evidence of response and progression on computed tomography and bone scan and with the degree of copper depletion measured by serum ceruloplasmin level. The antiangiogenic effects of therapy will also be evaluated by measurement of serum VEGF, bFGF, and interleukins 6 and 8 which are associated with cell proliferation and bone metabolism. The levels of these growth factors/cytokines will then be correlated with the degree of copper depletion, PSA level and response to therapy.

[unreadable] DESCRIPTION (provided by applicant): [unreadable] The proposed research examines the learning and memory functions of limbic brain circuitry involving the hippocampus, retrosplenial cortex, and the anterior and lateral dorsal thalamus. Pathology in these regions has been associated with the human cognitive impairments seen in amnesic syndromes and Alzheimer's disease. The specific goal is to examine the contributions these regions make to contextual learning. It is proposed that these regions function cooperatively to encode the cognitive and behavioral features of a learning context and thereby provide a basis for determining appropriate behaviors in different learning situations. To assess this, multi-site neuronal recording and temporary inactivation techniques will be employed while rats learn to obtain food rewards in two different learning contexts. [unreadable] [unreadable]

[unreadable] DESCRIPTION (provided by applicant): Career Development Aim: To develop into an independent, productive academician conducting translational research in the field of HIV pathogenesis. Career Development Methods: An individualized, mentored curriculum of peer and faculty interactions, formal didactics, and study development are proposed. Scientific Background: 15,000 people in the world will become infected with human immunodeficiency virus (HIV) today. Most of these infections will occur through sexual contact, and yet HIV within genital secretions is not as well characterized as that found in the blood. Genital tract compartmentalization has profound consequences on the development of drug resistance and the selection of HIV quasispecies for transmission. Research Aim: To characterize the genital tract as a separate compartment than from the blood secondary to different host cell selection immunologic responses and pharmacologic penetration. [unreadable] [unreadable] Research Methods: Male genital secretions are a complex mixture of secretions and cells. The amount of HIV within male genital secretions is highly variable. Sequestration of HIV within the genital tract will be explored by studying the role of the prostate, which is known to sequester both bacterial and fungal pathogens and is a source of seminal fluid. This will be investigated by examining the effect of prostate massage, which increases the amount of prostatic fluid in genital secretions, on the amount of cell-free and cell associated virus. Longitudinally collected paired samples of genital secretions and blood from subjects enrolled in the UCSD Acute and Early HIV Cohort will be examined. From these samples HIV RNA will be extracted and examined with clonal and consensus sequencing and length polymorphism detection of the HIV env and gag coding regions. Comparing the genetic diversity of HIV between the genital tract and blood and subsequent divergence will give a better idea of the type of compartmentalization that is occurring and what kind of virus is ultimately being transmitted. Using these same samples the appearance and disappearance of drug resistance will be tracked. This may shed light on the mechanism of drug resistance development and why so many new infections are occurring with drug resistant HIV. Significance: These studies are important to characterize the compartmentalization of HIV in the genital tract, and to understand factors affecting the transmission of HIV and treatment approaches to minimize drug resistance. [unreadable] [unreadable]

The Mathematics and Science Partnership of Greater Philadelphia (MSPGP) focuses on improving secondary mathematics and science, grades 6-12, in an "open loop" environment typical of sprawling, densely-populated greater metropolitan areas containing hundreds of school districts and dozens of institutions of higher education. In an effort to build the required linkages that address issues such as bringing strong local successes to scale and preventing program erosion over time, the MSPGP brings together as core partners 46 school districts and 13 institutions of higher education in eight Pennsylvania and four New Jersey counties in the region outside of Philadelphia. The MSPGP model includes a "Core Connector" organizational structure that provides a way to facilitate and grow partnerships between grades 6-12 teachers and administrators and faculty from higher educational institutions. To assess the progress of mathematics and science programs and college/university preservice programs, the MSPGP uses five-stage "On the Road to Reform" rubrics and customizes project activities to each circumstance depending on where any particular partner is on the journey.

DESCRIPTION (provided by applicant): It would be difficult to overstate the magnitude of the burden depressive disorders pose, not only to society and to the friends and relatives of those afflicted, but especially to depressed people themselves. Marital discord is a substantial risk factor for major depression, and this investigation is designed to clarify the nature of the association between depression and discord through detailed examination of inter-spousal criticism. The purpose is two-fold. First of all, this investigation seeks to determine whether perceptions of excessive spousal criticism are due to depressed people's biased perceptions of otherwise benign communications or to their accurate perceptions of comments from genuinely hypercritical spouses. Secondly, this study seeks to clarify and contextualize various individual differences predisposing to sending and receiving criticism. Carefully diagnosed depressed and non-depressed married people and their spouses will participate in a laboratory session during which they each complete marital, symptom, and personality questionnaires as well as undertaking a dyadic criticism perception accuracy task. The stimulus interaction for the criticism accuracy task will be a social support discussion of something one spouse would like to change about themselves. Both spouses then review the interaction videotape. The target spouses indicate time-linked thoughts and feelings they recall having experienced during the original interaction. Non-target ("source") partners review these same moments and indicate the thoughts and feelings they perceived their spouse as having experienced as well as disclosing their own intentions as they recall having them during the original interaction. Signal detection analyses will be deployed to determine the accuracy and bias in inferring criticism as evidenced by discrepancies between the two spouses' reports. Outside observers will also provide criticality ratings for comparison purposes. The role of individual differences in perceiving and emitting criticism will also be examined.

CLINICAL TRIALS OFFICE The Clinical Trials Office (CTO) provides infrastructure support for the conduct of clinical trials at the University of Michigan Comprehensive Cancer Center. CTO services are grouped into three major categories;regulatory services, data management services, and information technologies services. Major areas of responsibility include support for regulatory submissions to the IRB, two Protocol Review Committees (therapeutic and prevention), GCRC and other institutional review committees, FDA, and NCI. The CTO provides data management services for PRC-approved clinical trials as well as serving as a centralized data repository for research data generated on clinical trials. These services include study implementation, data collection and entry, and submission to investigators and outside sponsors. The information technology services include Case Report Form (CRF) development and database implementation and maintenance. The primary mission of the CTO is to assist the Cancer Center investigators in the development, conduct, and reporting of innovative clinical research in an efficient, regulatory compliant, and scientifically sound manner.

DESCRIPTION (provided by applicant): HIV-1 is one of the most genetically diverse pathogens on earth. In fact, some coding regions, such as envelope, can have more than 30% genetic difference between clades. The neurocognitive effects of clade B HIV-1 infection have been well characterized in the developed world;however, there continues to be controversy surronding the effect of non-clade B infection on the neurocognitive functioning. The overarching aims of the proposed study is to 1) characterize and compare the burden of HIV associated neurocognitive disorders (HAND) occurring among individuals living in Brazil, China, India, Romania and the United States and infected with CRF01_AE and clades B, C and F, and 2) evaluate the neurovirulent and neurotropic genotypic determinants between subtypes. Important research questions that remain unanswered include: How do the prevalence, nature and course of HAND in various parts of the world compare when analyzed by population and subtype? What specific viral genetic characteristics are associated with neurovirulence and seeding of the central nervous system (CNS)? The current proposal is designed to systematically address these issues by: 1) determining the differential effect of HIV subtype on neurocognitive performance by measuring HAND using standardized measures in previously established cohorts in Brazil (clades B and C), China (CRF01_AE and clades B and C), India (clade C), Romania (clade F) and the United States (clade B), 2) determining viral genetic motifs from HIV RNA that are conserved during HAND by subtype and by study population by performing clade-typing of study participants by generating population based sequences of the env and tat coding regions from HIV RNA and DNA extracted from blood, and 3) determining viral genetic motifs from HIV RNA that are conserved during CNS compartmentalization between clades B and C by performing clonal sequencing of the env and tat coding regions from HIV RNA extracted from paired blood and CSF samples collected from participants with clade B or C infection in Brazil, India and the United States. The comparison of the burden of HAND between groups infected with different HIV clades requires standardized procedures for the measurement of HAND within and across populations. Investigations into clade-specific genotypic determinants of HIV neuropathogenesis and neurotropism requires: 1) well-characterized study populations and biologic samples from study participants, 2) standardization of measurements of neurocognitive functioning, 3) high quality HIV RNA sequencing capability in all research study settings, 4) secure and reliable data management and communication capabilities for sequence and study data between participating sites, and 5) expertise in state-of- the-art genotypic analysis. Our group has demonstrated experience in each of these areas.

DESCRIPTION (provided by applicant): When antiretroviral therapy (ART) fails to suppress HIV replication, drug resistance can develop and be transmitted. This can be particularly devastating in resource limited settings where only a few ART regimens are available. The overarching aims of this proposal are to enhance a productive collaboration between UCSD and investigators in Chennai, India and to evaluate unique cost-containing methods for viral load monitoring of Indian patients on first-line ART. Specifically, we will enroll 500 HIV-infected patients followed at a HIV clinic in Chennai, India for monitoring of virologic failure of first-line ART. We will evaluate three approaches of nucleic acid testing (NAT) for HIV RNA: 1) individual samples, 2) minipools of 5 samples, and 3) 10x10 sample pooling matrix. Innovatively, the pooling methods make use of the quantitative information that is available from the viral loads to identify individual samples with virologic failure within the platforms of pooled samples. We will define the test characteristics of each method including: accuracy, efficiency, result turn-around time and costs, and we will determine if simple adherence measures can improve these test characteristics. These experiments will also allow us to define the prevalence of virologic failure in this study population group. Taken together, the generated data will serve as the basis for choosing a sustainable method for monitoring for virologic failure in Southern India. PUBLIC HEALTH RELEVANCE: We still struggle with availability of effective antiretroviral therapy and CD4 monitoring in most parts of the world where most of the people infected with HIV live. In these settings, the availability of viral load monitoring is practically unattainable for the vast majority of patients, often because of the lack of infrastructure and technical expertise. Even when infrastructure and expertise exist, cost prohibits its use. We propose in this application to evaluate the test characteristics of novel methods that may ultimately bring viral load monitoring to resource-limited settings.

The goal of this project is to integrate subject data from local research, clinical and public health entities that are screening for and treating HIV infected individual. Data from these sources will be obtained in a de?identified manner, parsed and then organized into a HIPAA compliant database containing socio?demographic, geographic and phylogenetic information for each subject. The database will be set up to update itself in real?time as new HIV infections are identified in our catchment area. In addition, we will use the viral sequence data to map out the phylogenetic network structure of our local epidemic. We will utilize a background of nationwide sequences obtained HIV sequence repositories to improve the signal in our phylogenetic structures, and use Bayesian maximum likelihood analysis to build these networks in a robust manner, leveraging the bioinformatics expertise at our institution, the University of California, San Diego. We will also update this network structure in a real?time fashion as new infections are identified. Finally we will use this constructed surveillance system to map the socio?demographic, geographic and phylogenetic locations of newly identified acute and early HIV infections, and use this information to direct community specific prevention resources (i.e. needle exchange, education resources etc?) with the ultimate goal of preventing HIV transmission clusters from developing or expanding.

Project Summary This application is a proposed renewal of Dr. Davey Smith's K24 grant entitled `Mentoring Translational Research in HIV'. This grant supported the training of over 30 trainees and 80 publications to date. While significant progress in HIV research continues to be made, it is not likely that a safe and effective cure or vaccine for HIV will be developed soon. The success of these goals will depend on future HIV researchers, many of whom are likely to be trained in the next five years. The goal of this K24 proposal is to provide Dr. Smith with sufficient support to mentor trainees in state-of-the-art translational and patient oriented HIV research and to expand his involvement in UCSD's wide array of training programs. This application is built upon funded and ongoing research projects, where the mentee is the primary leader and is trained in every aspect of study design, execution, analyses, results interpretation and publication. The presented studies provide opportunities for in-depth mentoring as well as investigation into important and open scientific questions in HIV pathogenesis. These goals will be met in the following three aims with a core group of junior and senior trainees. SPECIFIC AIM 1: To determine how co-infections impact HIV pathogenesis. SPECIFIC AIM 2: To develop and validate new molecular virology technology. SPECIFIC AIM 3: To use molecular epidemiologic methods to inform HIV prevention.

DESCRIPTION (provided by applicant): HIV-1 is one of the most genetically diverse pathogens on earth. In fact, some coding regions, such as envelope, can have more than 30% genetic difference between clades. The neurocognitive effects of clade B HIV-1 infection have been well characterized in the developed world; however, there continues to be controversy surronding the effect of non-clade B infection on the neurocognitive functioning. The overarching aims of the proposed study is to 1) characterize and compare the burden of HIV associated neurocognitive disorders (HAND) occurring among individuals living in Brazil, China, India, Romania and the United States and infected with CRF01_AE and clades B, C and F, and 2) evaluate the neurovirulent and neurotropic genotypic determinants between subtypes. Important research questions that remain unanswered include: How do the prevalence, nature and course of HAND in various parts of the world compare when analyzed by population and subtype? What specific viral genetic characteristics are associated with neurovirulence and seeding of the central nervous system (CNS)? The current proposal is designed to systematically address these issues by: 1) determining the differential effect of HIV subtype on neurocognitive performance by measuring HAND using standardized measures in previously established cohorts in Brazil (clades B and C), China (CRF01_AE and clades B and C), India (clade C), Romania (clade F) and the United States (clade B), 2) determining viral genetic motifs from HIV RNA that are conserved during HAND by subtype and by study population by performing clade-typing of study participants by generating population based sequences of the env and tat coding regions from HIV RNA and DNA extracted from blood, and 3) determining viral genetic motifs from HIV RNA that are conserved during CNS compartmentalization between clades B and C by performing clonal sequencing of the env and tat coding regions from HIV RNA extracted from paired blood and CSF samples collected from participants with clade B or C infection in Brazil, India and the United States. The comparison of the burden of HAND between groups infected with different HIV clades requires standardized procedures for the measurement of HAND within and across populations. Investigations into clade-specific genotypic determinants of HIV neuropathogenesis and neurotropism requires: 1) well-characterized study populations and biologic samples from study participants, 2) standardization of measurements of neurocognitive functioning, 3) high quality HIV RNA sequencing capability in all research study settings, 4) secure and reliable data management and communication capabilities for sequence and study data between participating sites, and 5) expertise in state-of- the-art genotypic analysis. Our group has demonstrated experience in each of these areas.

DESCRIPTION (provided by applicant): HIV-associated neurocognitive disorder (HAND) continues to be a debilitating disorder affecting the aging HIV patient population. This is despite the fact that currently available highly active antiretroviral therapy (HAART) is capable of suppressing HIV RNA to undetectable levels in both blood and cerebrospinal fluid (CSF). The persistence of HAND is likely due to a complex interplay between poorly understood virologic factors and natural aging processes. Understanding the biological determinants at the core of this complex interplay, and how they lead to the development of HAND even in the presence of suppressive HAART, will require a more detailed investigation of virologic factors and how they compound various human aging pathways. To better understand the biological correlates of HAND, we propose to use an analysis that combines clinical, neuromedical, and laboratory data with ultradeep sequencing (UDS) technology. We have identified a group of 56 individuals from the CNS HIV Antiretroviral Therapy Effects Research (CHARTER) cohort who are older than 50 years in age, have been followed longitudinally for at least four years, and have been suppressed with HAART throughout this time. Using UDS, we propose to sequence the env, gag, pro and reverse transcriptase coding regions of HIV-1 DNA populations in peripheral blood mononuclear cells (PBMCs) and CSF cell pellets (approximately 400 samples and 6.5 million sequences in total). These data will then be co-analyzed with clinical, antiretroviral, host geneti, and bio-marker data currently available in CHARTER. Specifically, we will investigate how the age of a subject and the presence or absence of HAND are associated with HIV-1 DNA levels in PBMCs and CSF cell pellets (Aim 1), the frequency of viral populations with differing co-receptor usage and drug resistance (Aim 2), anatomic compartmentalization and viral diversity between blood and CSF (Aim 3), and the rate of viral evolution during HAART (Aim 4). These analyses will be accomplished using a combination of phylogenetic, computational, and statistical techniques we developed and employ as part of our research at UCSD to better understand aspects of viral dynamics within human hosts. Collectively, this study will yield a more complete understanding of how various virologic factors impact natural aging processes and thereby lead to HAND within the context of HAART.

No abstract provided

DESCRIPTION (provided by applicant): The goal of this proposal is to develop and validate novel compressed sensing (CS) approaches to dramatically improve the spatial and temporal resolution of quantitative dynamic contrast enhanced magnetic resonance imaging (DCE-MRI). CS exploits prior information (assumptions) about MR images to infer missing data and produce high-quality images from significantly less data than previously thought possible. CS has already proven extremely successful in MR angiography and cardiac MRI, where it has accelerated some acquisitions by up to 10- to 100-fold, but is relatively unexplored in cancer imaging. DCE-MRI involves the serial acquisition of heavily T1-weighted images before and after the injection of a contrast agent to increase water relaxation rates in tissues. The resultin data can then be analyzed with appropriate pharmacokinetic models to extract quantitative parameters reporting on, for example, vessel perfusion and permeability, and tissue volume fractions. DCE-MRI has been applied to predict the early response to neoadjuvant chemotherapy in breast cancer, but the technique is not yet robust and accurate enough for the clinic. A fundamental practical limitation of DCE-MRI is the necessity to simultaneously acquire high temporal resolution, to adequately sample the contrast time course, and high spatial resolution, which is required for clinical morphological assessment and accurate tumor delineation. In traditional Cartesian MRI acquisitions, one must choose between high spatial or high temporal resolution before the scan. With a golden ratio acquisition, the tradeoff between spatial and temporal resolution is eliminated. A single DCE-MRI scan may then be used for both accurate kinetic modeling by slicing the data at high temporal cadence, while also allowing a high spatial resolution image to be formed by taking the data as a whole. Thus, a golden ratio acquisition coupled with CS has great potential to enable a clinically relevant DCE-MRI protocol that provides adequate temporal resolution for kinetic modeling without sacrificing the spatial resolution required for morphological evaluation. This project has three aims: (1) to develop a compressed sensing based high temporal resolution protocol for quantitative DCE-MRI, (2) to develop a compressed sensing based high spatial resolution T1-weighted anatomical imaging protocol for morphological evaluation, and (3) to apply the developed CS-based protocols in vivo for validation and evaluation. If this project is successful, it will significantly improve te ability to predict response to neoadjuvant chemotherapy, provide new CS methods for the community to apply to other in vivo applications, and validate CS in an important cancer imaging application.

DESCRIPTION (provided by applicant): We have made great strides to reach the modern era of antiretroviral therapy (ART) in resource wealthy settings. We now know that optimal clinical outcomes require maximal suppression of viral replication by combination ART to maintain the integrity of the immune system and to avoid the selection of drug resistant HIV. We also know that drug resistance (DR) represents a real public health threat since it can be spread to others thereby limiting the usefulness of certain ART regimens to them. Our current standard clinical practices in resource wealthy settings include starting combination ART when clinically appropriate, regular clinical, viral load and CD4+ count monitoring, and DR testing when viral replication is detected in the setting of ART exposure to inform the selection of appropriate second and third line ART regimens. This same standard of HIV care is not currently available in many resource limited settings. In many regions, struggles continue to provide access to combination ART and CD4 count monitoring where most of the people infected with HIV live. In these settings, the availability of viral load monitoring and DR testing for all practical purposes remains unattainable for the vast majority of patients, related to the lack of infrastructure and technical expertise in many instances. Even when infrastructure and expertise exist, cost often prohibits their use. We feel that it is imperative and urgent to overcome the obstacles to provide optimal HIV care, including virologic monitoring and DR testing, for patients receiving ART in resource limited settings. This will not only improve the standard of care for individual patients but also reduce the transmission of HIV DR. In the absence of viral load monitoring and DR testing a secondary epidemic of drug resistant HIV may proceed unchecked with disastrous public health consequences. To this end, we propose a method that may overcome some of these obstacles and ultimately bring virologic monitoring and DR testing to resource limited settings in a potentially cost effective and sustainable manner. In this study, we will compare the 'gold standards' of (i) individual viral load testing and drug resistance genotyping for samples found to virologic failure to (ii) an assay that combines nucleic acid testing and DR genotyping (NAT+DR assay). This NAT+DR assay combines dilution techniques via sample pooling, PCR amplification of the HIV-1 reverse transcriptase (RT), deconvolution algorithms to identify patients with virologic failure, and sequencing of PCR product to determine drug resistance. To evaluate this method, we will enroll 500 HIV-infected patients followed at YRG CARE on first-line ART and screen baseline and six month samples with both individual viral loads and the new NAT+DR assay. We will then evaluate the NAT+DR assay based on accuracy, efficacy, turnaround time of results and costs. We have proposed a quick start and aggressive timeline for this project because we feel that the rapid validation of these methods could be crucial for the health of many living with HIV in the world. PUBLIC HEALTH RELEVANCE: We still struggle with availability of combination ART and CD4 monitoring in most parts of the world where most of the people infected with HIV live, including India. In these settings, the availability of virologic monitoring and drug resistant testing is practically unattainable for the vast majority of patients, which is often because of the lack of infrastructure and technical expertise, but even when infrastructure and expertise exist, cost prohibits its use. We propose in this application a method that may ultimately bring virologic monitoring and drug resistance testing to resource limited settings.

? DESCRIPTION: This study is designed to meet the goals of the RFA-DA-15-018: Developing Technologies and Tools to Monitor HIV Brain Reservoirs and How They May be Altered by Exposure to Substances of Abuse. To meet this challenge in the R21 phase, we propose to develop and validate a combined next generation sequencing (Pacific Biosciences single-molecule, real time (SMRT) sequencing) and bioinformatics platform to accurately measure and characterize HIV DNA populations in brain and lymph node tissues. In particular, this platform will be able to measure and identify proviral populations that are (i) intact and replication competent, (ii) intact but replication incompetent, and (iii) not intact. We will develop and validate the proposed platform based on international standard procedures for diagnostic molecular methods (i.e. CAP and OIE) using brain and lymph node tissues from HIV-infected individuals on optimized antiretroviral therapy for at least 2 years before death. Once developed and validated, we will use this technology in the R33 phase to determine how HIV DNA populates the brains of stimulant users and non-users.

? DESCRIPTION (provided by applicant): This study is designed to meet the goals of the RFA Innovative assays to quantify the latent HIV reservoir that can facilitate proof-of-concept studies for curing HIV infection. To meet this challenge we propose to develop and validate a combined next generation sequencing (Pacific Biosciences single-molecule, real time (SMRT) sequencing) and bioinformatics platform to accurately measure replication-competent provirus (i.e. Proviral Phenotypic Predictor; P3). An over-arching objective would be for the assay to have a higher throughput, faster turnaround, lower cost, and greater reproducibility than the current standard quantitative viral outgrowth assay (QVOA). We will develop and validate the proposed P3 platform based on international standard procedures for diagnostic molecular methods (i.e. CAP and OIE) using latently infected cells from HIV-infected individuals on optimized antiretroviral therapy for at least 2 years. These participants will be well-characterize to estimated duration of infection at the start of their therapy, and will represent high, medium and low levels of HIV DNA levels. Such characterizations will be necessary to adequately address the dynamic range of the proposed P3 platform. Once P3 methods have been optimized, we will rigorously compare P3 and QVOA methods in their ability to characterize the replication competent proviral reservoir and by costs and turnaround time for results.

NEUROGERM CORE PROJECT SUMMARY/ABSTRACT: The overall NeuroGerm (NG) Core is co-directed by Drs. Davey Smith and Rob Knight and comprises a ?Virology-Inflammation Unit? and a ?Microbiome Unit?. These units will be integral to the HIV Neurobehavioral Research Center (HNRC), and will develop new knowledge and new technologies to meet the Center's scientific and training goals. Specifically, the units will provide HNRC and collaborating researchers with expertise and techniques relevant to the virologic and immunologic investigation of HIV-associated neurocognitive disorder (HAND) with an enhanced focus on the role of the microbiome, and on HIV eradication from the central nervous system (CNS). The NG Core will offer a range of services focused on: (i) assay development, validation and provision, (ii) analytical support, (iii) quality assurance, (iv) training, and (v) provision of samples and reagents. Since viral populations in the central nervous system (CNS) can be genetically distinct from populations in other tissues, such as blood, lymphoid and genital tract, advanced virologic techniques will be needed to characterize neurotropism and neurovirulence in the CNS in relation to HAND, as well as to design appropriate HIV eradication strategies involving the CNS. The NG Core will also enhance the HNRC's transdiciplinary focus through close collaboration with the NeuroBiology, NeuroAssessment and Developmental Cores, utilizing multiple laboratory and analytical techniques specific to CNS derived virus, and characterizing the microbiome in relation to neurocognitive functioning. In summary, the NG Core will provide the proposed quality services to tackle the mission of the HNRC. The provision of these services will be based on a detailed strategic planning process designed to maintain flexibility and to maximize utility of offered services based on the scientific needs of the field and our associated HIV investigators. With its variety of cutting-edge molecular and immunologic techniques, the NG core will be crucial to the understanding and mechanisms of the neuropathogenesis of HIV.

? DESCRIPTION (provided by applicant): Most recent HIV cure efforts have focused on strategies to induce virus from HIV-infected cells during antiretroviral therapy (ART), thus leaving the cellular reservoir vulnerable to the host immune system, while ART prevents new cells from being infected. Unfortunately, such efforts have not demonstrated much activity. On the other hand, it has been documented for almost two decades that routine clinical vaccines can induce viral production from infected cells, even during ART, and that these levels of induction are much higher than those seen by recently described interventions (e.g. histone deacetylase inhibitors [HDACi], IL-7, disulfram). Therefore, based on newly generated preliminary data, we propose a randomized clinical trial to determine how two commonly used vaccines (against Influenza and Pneumococcus) can stimulate the immune system and induce HIV transcription in the setting of ART. Specifically, we propose a randomized cross-over controlled trial where 56 participants will receive one injection every three months (Influenza, Pneumococcal, and Placebo) in random order with multiple specimen collections after each injection. The primary objective of this study will be to determine if participants have a higher absolute increase in levels of cell-associated HIV RNA transcription after receiving active vaccines when compared placebo injection. Secondary objectives will be to determine if these vaccines also: (i) induce HIV transcription selectively or non-selectively (as evaluated by panmixis tests between sequences obtained from cellular HIV DNA and RNA populations), (ii) influence total HIV DNA levels, (iii) influence fraction of replication competent proviral levels (s evaluated by quantitative viral outgrowth assays), (iv) stimulate generalized and lymphocyte immune activation, and (v) stimulate vaccine- and HIV- specific immune responses. To best delineate these possible effects, we will also measure other sources of incidental immune stimulation, like shedding of herpesviruses, microbial translocation and incident illnesses.

PROJECT 2. LATE TREATMENT ABSTRACT: Blood and Tissues Reservoirs Most human studies have characterized circulating reservoirs and some non-blood HIV reservoirs using limited biological samples from otherwise healthy HIV-infected individuals, like genital sections, gut biopsies and cerebrospinal fluid, or autopsy specimens with little pre-mortem characterization. Therefore, our understanding of how HIV populates tissues throughout the body and how these populations are associated with each other- remains limited. Our proposed Revealing Reservoirs during Rebound (R3) program will take the next steps in finding a cure for HIV by understanding how HIV persists in and populates reservoirs throughout the body. Specifically, the Late Treatment Research Project (RP) will maximize data and samples collected from a `Last Gift' cohort of HIV-infected terminally-ill individuals. These altruistic individuals will provide: (i) detailed clinical, risk and socio-demographic information before their death; (ii) weekly blood collections while they are alive, before and after voluntarily stopping ART and, (iii) their entire bodies after they die. This cohort will allow us to characterize the HIV populations in blood before ART is stopped and during rebound viremia, and throughout the body after death. To complement studies proposed in the Early Treatment RP, this RP will determine: Aim 1: Virologic and immunologic quantities measured during ART that predict dynamics of viral rebound dynamics when ART is stopped (timing, size, slope); Aim 2: Dynamics of HIV variants populating CD4+ T cell subsets and in various anatomic tissues after ART interruption. Aim 3: Determine immunologic mechanisms associated with HIV persistence during ART and after ART interruption. By investigating HIV-infected individuals who started ART during chronic infection and thus have larger HIV reservoirs and more immune damage from HIV, the Late Treatment RP will complement the Early Treatment RP, which will investigate persons who started ART during acute and early infection, and thus have smaller HIV reservoirs and more intact immune systems. This RP will also create new knowledge about how HIV resides in circulating CD4+ T cells and in anatomic tissues, and these data will be analyzed in conjunction with data from the Early Treatment RP vianew analytical methods with the Quantitative Methods RP to more fully understand HIV dynamics throughout the body.

Project Summary/Abstract Core K - Modern HIV research efforts require reliable links between clinical and basic science investigators. These links include characterization and access to clinical specimens, laboratory based assays designed for clinical specimens, interpretation and management of generated data, and training in these areas. The Translational Virology (TV) Core of the San Diego (SD) CFAR is designed to be this reliable and cost-effective link that provides added value to HIV research within our Center. The TV Core accomplishes this goal for a wide range of HIV research-related activities involving emerging and senior investigators, established and burgeoning programs, clinical and basic science investigators, domestic and international efforts, NIH and other funders, etc. Over the past 4 years, the TV Core has provided services to 199 investigators, contributed to 168 publications, supported investigators funded by 14 NIH institutes, and trained 127 individuals in safe laboratory procedures for working with HIV. Specifically, the TV Core provides services through four defined units that work closely with other CFAR Cores, as outlined: (1) Specimen Unit: obtains, processes, tracks, ships and cryostores samples. (Primary collaborating Core: Clinical Investigation Core). (2) Assay Unit: develops and provides a wide-range of clinically-relevant assays (HIV genotyping, viral loads, antibody and nucleic acid detection, etc.). (Primary collaborating Core: Genomics and Sequencing). (3) Quality Control Unit: provides quality control procedures for domestic and international CFAR-affiliated investigators, and analytical support for generated data. (Primary collaborating Core: BAM Cores). (4) Training Unit: provides training and certification in BL2 and BL3 safety and procedures, specimen processing, PCR, genotyping techniques, and laboratory quality assurance. (Primary collaborating Cores: Developmental and International Cores).