Meningitis - A Medical Dictionary, Bibliography, and Annotated Research Guide to Internet References

MENINGITIS A M EDICAL D ICTIONARY , B IBLIOGRAPHY , AND A NNOTATED R ESEARCH G UIDE TO I NTERNET R E FERENCES

J AMES N. P ARKER , M.D. AND P HILIP M. P ARKER , P H .D., E DITORS

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ICON Health Publications ICON Group International, Inc. 4370 La Jolla Village Drive, 4th Floor San Diego, CA 92122 USA Copyright 2003 by ICON Group International, Inc. Copyright 2003 by ICON Group International, Inc. All rights reserved. This book is protected by copyright. No part of it may be reproduced, stored in a retrieval system, or transmitted in any form or by any means, electronic, mechanical, photocopying, recording, or otherwise, without written permission from the publisher. Printed in the United States of America. Last digit indicates print number: 10 9 8 7 6 4 5 3 2 1

Publisher, Health Care: Philip Parker, Ph.D. Editor(s): James Parker, M.D., Philip Parker, Ph.D. Publisher's note: The ideas, procedures, and suggestions contained in this book are not intended for the diagnosis or treatment of a health problem. As new medical or scientific information becomes available from academic and clinical research, recommended treatments and drug therapies may undergo changes. The authors, editors, and publisher have attempted to make the information in this book up to date and accurate in accord with accepted standards at the time of publication. The authors, editors, and publisher are not responsible for errors or omissions or for consequences from application of the book, and make no warranty, expressed or implied, in regard to the contents of this book. Any practice described in this book should be applied by the reader in accordance with professional standards of care used in regard to the unique circumstances that may apply in each situation. The reader is advised to always check product information (package inserts) for changes and new information regarding dosage and contraindications before prescribing any drug or pharmacological product. Caution is especially urged when using new or infrequently ordered drugs, herbal remedies, vitamins and supplements, alternative therapies, complementary therapies and medicines, and integrative medical treatments. Cataloging-in-Publication Data Parker, James N., 1961Parker, Philip M., 1960Meningitis: A Medical Dictionary, Bibliography, and Annotated Research Guide to Internet References / James N. Parker and Philip M. Parker, editors p. cm. Includes bibliographical references, glossary, and index. ISBN: 0-597-83628-0 1. Meningitis-Popular works. I. Title.

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Disclaimer This publication is not intended to be used for the diagnosis or treatment of a health problem. It is sold with the understanding that the publisher, editors, and authors are not engaging in the rendering of medical, psychological, financial, legal, or other professional services. References to any entity, product, service, or source of information that may be contained in this publication should not be considered an endorsement, either direct or implied, by the publisher, editors, or authors. ICON Group International, Inc., the editors, and the authors are not responsible for the content of any Web pages or publications referenced in this publication.

Copyright Notice If a physician wishes to copy limited passages from this book for patient use, this right is automatically granted without written permission from ICON Group International, Inc. (ICON Group). However, all of ICON Group publications have copyrights. With exception to the above, copying our publications in whole or in part, for whatever reason, is a violation of copyright laws and can lead to penalties and fines. Should you want to copy tables, graphs, or other materials, please contact us to request permission (E-mail: [email protected]). ICON Group often grants permission for very limited reproduction of our publications for internal use, press releases, and academic research. Such reproduction requires confirmed permission from ICON Group International Inc. The disclaimer above must accompany all reproductions, in whole or in part, of this book.

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Acknowledgements The collective knowledge generated from academic and applied research summarized in various references has been critical in the creation of this book which is best viewed as a comprehensive compilation and collection of information prepared by various official agencies which produce publications on meningitis. Books in this series draw from various agencies and institutions associated with the United States Department of Health and Human Services, and in particular, the Office of the Secretary of Health and Human Services (OS), the Administration for Children and Families (ACF), the Administration on Aging (AOA), the Agency for Healthcare Research and Quality (AHRQ), the Agency for Toxic Substances and Disease Registry (ATSDR), the Centers for Disease Control and Prevention (CDC), the Food and Drug Administration (FDA), the Healthcare Financing Administration (HCFA), the Health Resources and Services Administration (HRSA), the Indian Health Service (IHS), the institutions of the National Institutes of Health (NIH), the Program Support Center (PSC), and the Substance Abuse and Mental Health Services Administration (SAMHSA). In addition to these sources, information gathered from the National Library of Medicine, the United States Patent Office, the European Union, and their related organizations has been invaluable in the creation of this book. Some of the work represented was financially supported by the Research and Development Committee at INSEAD. This support is gratefully acknowledged. Finally, special thanks are owed to Tiffany Freeman for her excellent editorial support.

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About the Editors James N. Parker, M.D. Dr. James N. Parker received his Bachelor of Science degree in Psychobiology from the University of California, Riverside and his M.D. from the University of California, San Diego. In addition to authoring numerous research publications, he has lectured at various academic institutions. Dr. Parker is the medical editor for health books by ICON Health Publications. Philip M. Parker, Ph.D. Philip M. Parker is the Eli Lilly Chair Professor of Innovation, Business and Society at INSEAD (Fontainebleau, France and Singapore). Dr. Parker has also been Professor at the University of California, San Diego and has taught courses at Harvard University, the Hong Kong University of Science and Technology, the Massachusetts Institute of Technology, Stanford University, and UCLA. Dr. Parker is the associate editor for ICON Health Publications.

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About ICON Health Publications To discover more about ICON Health Publications, simply check with your preferred online booksellers, including Barnes & Noble.com and Amazon.com which currently carry all of our titles. Or, feel free to contact us directly for bulk purchases or institutional discounts: ICON Group International, Inc. 4370 La Jolla Village Drive, Fourth Floor San Diego, CA 92122 USA Fax: 858-546-4341 Web site: www.icongrouponline.com/health

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Table of Contents FORWARD .......................................................................................................................................... 1 CHAPTER 1. STUDIES ON MENINGITIS .............................................................................................. 3 Overview........................................................................................................................................ 3 The Combined Health Information Database................................................................................. 3 Federally Funded Research on Meningitis..................................................................................... 5 E-Journals: PubMed Central ....................................................................................................... 71 The National Library of Medicine: PubMed ................................................................................ 86 CHAPTER 2. NUTRITION AND MENINGITIS .................................................................................. 219 Overview.................................................................................................................................... 219 Finding Nutrition Studies on Meningitis ................................................................................. 219 Federal Resources on Nutrition ................................................................................................. 226 Additional Web Resources ......................................................................................................... 226 CHAPTER 3. ALTERNATIVE MEDICINE AND MENINGITIS ............................................................ 229 Overview.................................................................................................................................... 229 National Center for Complementary and Alternative Medicine................................................ 229 Additional Web Resources ......................................................................................................... 237 General References ..................................................................................................................... 238 CHAPTER 4. DISSERTATIONS ON MENINGITIS .............................................................................. 239 Overview.................................................................................................................................... 239 Dissertations on Meningitis ...................................................................................................... 239 Keeping Current ........................................................................................................................ 240 CHAPTER 5. CLINICAL TRIALS AND MENINGITIS ......................................................................... 241 Overview.................................................................................................................................... 241 Recent Trials on Meningitis ...................................................................................................... 241 Keeping Current on Clinical Trials ........................................................................................... 249 CHAPTER 6. PATENTS ON MENINGITIS ......................................................................................... 251 Overview.................................................................................................................................... 251 Patents on Meningitis................................................................................................................ 251 Patent Applications on Meningitis............................................................................................ 257 Keeping Current ........................................................................................................................ 258 CHAPTER 7. BOOKS ON MENINGITIS............................................................................................. 259 Overview.................................................................................................................................... 259 Book Summaries: Federal Agencies............................................................................................ 259 Book Summaries: Online Booksellers......................................................................................... 261 The National Library of Medicine Book Index ........................................................................... 263 Chapters on Meningitis ............................................................................................................. 264 CHAPTER 8. MULTIMEDIA ON MENINGITIS .................................................................................. 273 Overview.................................................................................................................................... 273 Video Recordings ....................................................................................................................... 273 Audio Recordings....................................................................................................................... 274 Bibliography: Multimedia on Meningitis .................................................................................. 275 CHAPTER 9. PERIODICALS AND NEWS ON MENINGITIS ............................................................... 277 Overview.................................................................................................................................... 277 News Services and Press Releases.............................................................................................. 277 Newsletter Articles .................................................................................................................... 283 Academic Periodicals covering Meningitis................................................................................ 285 CHAPTER 10. RESEARCHING MEDICATIONS................................................................................. 287 Overview.................................................................................................................................... 287 U.S. Pharmacopeia..................................................................................................................... 287 Commercial Databases ............................................................................................................... 288 Researching Orphan Drugs ....................................................................................................... 289

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APPENDIX A. PHYSICIAN RESOURCES .......................................................................................... 293 Overview.................................................................................................................................... 293 NIH Guidelines.......................................................................................................................... 293 NIH Databases........................................................................................................................... 295 Other Commercial Databases..................................................................................................... 298 APPENDIX B. PATIENT RESOURCES ............................................................................................... 299 Overview.................................................................................................................................... 299 Patient Guideline Sources.......................................................................................................... 299 Associations and Meningitis...................................................................................................... 305 Finding Associations.................................................................................................................. 307 APPENDIX C. FINDING MEDICAL LIBRARIES ................................................................................ 309 Overview.................................................................................................................................... 309 Preparation................................................................................................................................. 309 Finding a Local Medical Library................................................................................................ 309 Medical Libraries in the U.S. and Canada ................................................................................. 309 ONLINE GLOSSARIES................................................................................................................ 315 Online Dictionary Directories ................................................................................................... 319 MENINGITIS DICTIONARY ..................................................................................................... 321 INDEX .............................................................................................................................................. 419

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FORWARD In March 2001, the National Institutes of Health issued the following warning: "The number of Web sites offering health-related resources grows every day. Many sites provide valuable information, while others may have information that is unreliable or misleading."1 Furthermore, because of the rapid increase in Internet-based information, many hours can be wasted searching, selecting, and printing. Since only the smallest fraction of information dealing with meningitis is indexed in search engines, such as www.google.com or others, a non-systematic approach to Internet research can be not only time consuming, but also incomplete. This book was created for medical professionals, students, and members of the general public who want to know as much as possible about meningitis, using the most advanced research tools available and spending the least amount of time doing so. In addition to offering a structured and comprehensive bibliography, the pages that follow will tell you where and how to find reliable information covering virtually all topics related to meningitis, from the essentials to the most advanced areas of research. Public, academic, government, and peer-reviewed research studies are emphasized. Various abstracts are reproduced to give you some of the latest official information available to date on meningitis. Abundant guidance is given on how to obtain free-of-charge primary research results via the Internet. While this book focuses on the field of medicine, when some sources provide access to non-medical information relating to meningitis, these are noted in the text. E-book and electronic versions of this book are fully interactive with each of the Internet sites mentioned (clicking on a hyperlink automatically opens your browser to the site indicated). If you are using the hard copy version of this book, you can access a cited Web site by typing the provided Web address directly into your Internet browser. You may find it useful to refer to synonyms or related terms when accessing these Internet databases. NOTE: At the time of publication, the Web addresses were functional. However, some links may fail due to URL address changes, which is a common occurrence on the Internet. For readers unfamiliar with the Internet, detailed instructions are offered on how to access electronic resources. For readers unfamiliar with medical terminology, a comprehensive glossary is provided. For readers without access to Internet resources, a directory of medical libraries, that have or can locate references cited here, is given. We hope these resources will prove useful to the widest possible audience seeking information on meningitis. The Editors

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From the NIH, National Cancer Institute (NCI): http://www.cancer.gov/cancerinfo/ten-things-to-know.

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CHAPTER 1. STUDIES ON MENINGITIS Overview In this chapter, we will show you how to locate peer-reviewed references and studies on meningitis.

The Combined Health Information Database The Combined Health Information Database summarizes studies across numerous federal agencies. To limit your investigation to research studies and meningitis, you will need to use the advanced search options. First, go to http://chid.nih.gov/index.html. From there, select the “Detailed Search” option (or go directly to that page with the following hyperlink: http://chid.nih.gov/detail/detail.html). The trick in extracting studies is found in the drop boxes at the bottom of the search page where “You may refine your search by.” Select the dates and language you prefer, and the format option “Journal Article.” At the top of the search form, select the number of records you would like to see (we recommend 100) and check the box to display “whole records.” We recommend that you type “meningitis” (or synonyms) into the “For these words:” box. Consider using the option “anywhere in record” to make your search as broad as possible. If you want to limit the search to only a particular field, such as the title of the journal, then select this option in the “Search in these fields” drop box. The following is what you can expect from this type of search: •

Performance After Cochlear Implantation: A Comparison of Children Deafened by Meningitis and Congenitally Deaf Children Source: Journal of Laryngology and Otology. 114(1): 33-37. January 2000. Contact: Available from Royal Society of Medicine Press Limited. Publications Subscription Department, P.O. Box 9002, London W1A 0ZA, United Kingdom. E-mail: [email protected]. Summary: This article reports on a study in which the speech perception and speech production performance following cochlear implantation of congenitally deaf children and children deafened by meningitis were analyzed. Three groups consisting of 70 congenitally deaf children, 22 children deafened by meningitis before two years of age, and 14 children deafened by meningitis after two years of age were compared. The

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group deafened by meningitis after two years of age demonstrated significantly better speech perception than the other two groups. Their speech production appeared better, but did not achieve statistical significance compared with the other two groups. There were no significant differences in speech perception or production between the congenitally deaf group and the group deafened by meningitis before two years of age. Further research is required to determine whether this is a consequence of meningitis, whether they have failed to derive benefit from their previous auditory experience, or whether any benefit they may have derived has been lost during their period of deafness. 2 figures. 9 tables. 15 references. •

Risk Factors for Hearing Loss from Meningitis in Children Source: Archives of Otolaryngology-Head and Neck Surgery. 125(5): 509-514. May 1999. Contact: Available from American Medical Association. Subscriber Services, P.O. Box 10946, Chicago, IL 60610-0946. (800) 262-3250 or (312) 670-7827. Fax (312) 464-5831. Email: [email protected]. Website: www.ama-assn.org/oto. Summary: This article reports on a study undertaken to identify statistically significant risk factors for hearing loss in children with meningitis, determine the overall incidence of hearing loss in a large group of children with confirmed meningitis, and quantify the percentage of children with progressive or fluctuating hearing loss after meningitis. The study included 432 children admitted to the Children's Hospital, Birmingham, Alabama, from 1985 to 1995. Of 432 children with meningitis, 59 (13.7 percent) developed hearing loss. Of these 59 children, 46 (78.0 percent) had stable sensorineural hearing loss and 13 (22. 0 percent) had either progressive or fluctuating hearing loss. Of the variables examined, only 5 appeared to be significantly associated with the development of hearing loss: CT scan evidence of increased intracranial pressure, male sex, the common logarithm of glucose levels in the cerebrospinal fluid, Streptococcus pneumoniae as the causative organism, and the presence of neck rigidity. In the children with progressive hearing loss, the time for progression varied from 3 months to 4 years before hearing stabilized. 1 figure. 4 tables. 12 references. (AA-M).

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Prevention of Hearing Loss From Meningitis Source: Lancet. 350(9072): 158-159. July 19, 1997. Summary: This brief commentary presents information on the prevention of hearing loss due to meningitis. Before the introduction of the Haemophilus influenzae type b conjugate vaccines, bacterial meningitis accounted for close to 10 percent of acquired deafness in children in the United States. In hearing loss due to bacterial meningitis, suppurative labyrinthitis precedes the destruction of cochlear structures. Bacteria may reach the cochlea by various routes. Exactly how suppurative labyrinthitis leads to deafness is not totally clear. Hearing loss caused by bacterial meningitis occurs quite early in the course of the infection. Differences in the ability of various microorganisms to reach the cochlea and cause damage and in host response to infection are possible reasons why some children develop hearing loss after bacterial meningitis and some do not. In order to prevent or diminish the likelihood of hearing loss due to bacterial meningitis, the author recommends optimum antimicrobial therapy, surveillance and careful monitoring of recommended treatment regimens, dexamethasone as an adjunctive measure, and other, newer adjunctive drug therapies that may reduce the incidence of hearing loss. The author concludes that in developed countries, deafness due to H. influenzae type b meningitis has been all but eliminated by protein conjugate vaccines that prevent invasive infections effectively. The hope is that protein conjugate

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vaccines under development will eventually prevent pneumococcal and meningococcal infections, including meningitis. 1 figure. 7 references. •

Vestibular Dysfunction Due to Cryptococcal Meningitis Source: Otolaryngology-Head and Neck Surgery. 116(4): 536-540. April 1997. Contact: Available from Mosby-Year Book, Inc. Subscription Services, 11830 Westline Industrial Drive, St. Louis, MO 63146-3318. (800) 453-4351 or (314) 453-4351. Fax (314) 432-1158. E-mail: [email protected]. Summary: Cryptococcus neoformans is an encapsulated, opportunistic, fungal organism found primarily in soil and pigeon feces. Human beings presumably are infected through inhalation of the organism, which causes pulmonary (lung) cryptococcus that can then spread hematogenously (through the blood) to the central nervous system, bone, and kidneys. This article presents the case of a nonimmunosuppressed and previously healthy woman with subacute onset of severe ataxia (impaired ability to coordinate movement) in whom cryptococcal meningitis was subsequently diagnosed. This case is unique in that the patient had bilateral vestibular hypofunction and only a moderate unilateral sensorineural hearing loss (SNHL). The other unique aspect was the reversal of the vestibular symptoms after appropriate treatment, which included decompression of obstructive hydrocephalus (fluid on the brain) and long term antifungal therapy. This is the first case reported in the recent otolaryngologic literature in which a reversal rather than merely a stabilization of vestibular symptoms occurred in cryptococcal meningitis. The authors remind readers that the diagnosis of cryptococcal meningitis can be difficult because of the subacute nature of the infection and the initially mild symptoms. Audiovestibular damage is the result of direct invasion of the neural tissue and sensory end organs by the organism. 3 figures. 13 references.

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Hearing Impairment After Bacterial Meningitis: A Review Source: Archives of Disease in Childhood. 67(9): 1128-1133. 1992. Summary: This journal article, intended for health professionals, reviews the recent literature on the incidence of hearing impairment following bacterial meningitis. The article discusses the rate of hearing impairment following bacterial meningitis, possible predictive factors, the permanence of hearing loss following bacterial meningitis, and possible means of reducing the risks of postmeningitic sensorineural hearing loss. It recommends that all children recovering from bacterial meningitis be referred for audiological assessment before discharge from the hospital. Bibliographic references are included.

Federally Funded Research on Meningitis The U.S. Government supports a variety of research studies relating to meningitis. These studies are tracked by the Office of Extramural Research at the National Institutes of Health.2 CRISP (Computerized Retrieval of Information on Scientific Projects) is a searchable database of federally funded biomedical research projects conducted at universities, hospitals, and other institutions. 2

Healthcare projects are funded by the National Institutes of Health (NIH), Substance Abuse and Mental Health Services (SAMHSA), Health Resources and Services Administration (HRSA), Food and Drug Administration (FDA), Centers for Disease Control and Prevention (CDCP), Agency for Healthcare Research and Quality (AHRQ), and Office of Assistant Secretary of Health (OASH).

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Search the CRISP Web site at http://crisp.cit.nih.gov/crisp/crisp_query.generate_screen. You will have the option to perform targeted searches by various criteria, including geography, date, and topics related to meningitis. For most of the studies, the agencies reporting into CRISP provide summaries or abstracts. As opposed to clinical trial research using patients, many federally funded studies use animals or simulated models to explore meningitis. The following is typical of the type of information found when searching the CRISP database for meningitis: •

Project Title: 26TH INTL HERPESVIRUS WORKSHOP, REGENSBURG, GERMANY Principal Investigator & Institution: Nelson, Jay A. Director & Professor; Molecular Microbiol and Immun; Oregon Health & Science University Portland, OR 972393098 Timing: Fiscal Year 2001; Project Start 01-JUL-2001; Project End 01-AUG-2002 Summary: (provided by applicant) This proposal requests funds to enable young investigators to attend the 26th International Herpesvirus Workshop at the University of Regensburg in Germany, July 28-August 3, 2001. The International Herpesvirus Workshop is the premier scientific meeting for herpesvirus researchers, and the only meeting with an interdisciplinary focus on all the major subfamilies of herpesviruses and all aspects of research from molecular biology to clinical studies. The strength of the Workshop rests on the cross-fertilization that results from comparison of different herpesviruses, different approaches to key questions and on the support and participation of leading researchers in the field, most significantly including promising young investigators and students in training. Moreover, the forum is truly international, with broad-based world-wide attendance. The medical importance of this meeting is clearly indicated from the wide variety of diseases caused by the now recognized eight human herpesviruses. These include skin and eye ulcerations (HSV-1), genital lesions (HSV-2), meningitis and encephalitis (HSV-1 and HSV-2), infectious mononucleosis (EBV) chicken pox and shingles (VZV). CMV is a major cause of birth defects including mental retardation, blindness and deafness due to congenital transmission but also a significant opportunistic pathogen in AIDS patients and organ transplant recipients. More recently, CMV and HSV have been implicated as pathogenic contributors in the development of atherosclerosis. Cancer has also been associated with herpesvirus infections. EBV is associated with Burkitt's lymphoma, other B cell neoplasias and nasopharyngeal carcinoma. The most recent human herpesvirus discovered (HHV-8 or KSHV) is associated with Kaposi's sarcoma in AIDS patients and other immunosupressed persons and in other groups. All of the herpesviruses persist for life and therefore pose significant problems in the treatment of immunologically compromised persons. Disease due to reactivation of most human herpesviruses are a significant cause of morbidity and mortality in various immune patient populations. Shingles and post-herpetic neuralgia are problems in the elderly. Animal herpes significant economic importance to the poultry (Marek's and others), swine (pseudorabies virus), cattle (several bovine herpesviruses) and horses (several equine herpesviruses). In addition, these animal herpes serve as important model systems for studying herpesvirus pathogenesis. Finally, recombinant DNA technology permits the design of novel vaccines for controlling the spread of animal herpesvirus infection and the design of herpesvirus vectors for gene therapy. Workshop sessions will take an interdisciplinary approach to the following topics: virus structure, mechanism of virus entry and cell-cell spread, membrane proteins, pathogenesis and latency, DNA replication, vaccination and the immune response, transcriptional control, regulation of gene expression, chemotherapeutic targets, and virus gene therapy,

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Website: http://crisp.cit.nih.gov/crisp/Crisp_Query.Generate_Screen •

Project Title: A MENINGOCOCCAL LOS VACCINE Principal Investigator & Institution: Griffiss, John M. Associate Professor; Northern California Institute Res & Educ San Francisco, CA 941211545 Timing: Fiscal Year 2003; Project Start 15-JUN-2003; Project End 30-NOV-2005 Summary: (provided by applicant): Meningococcal disease primarily affects infants and very young children. The group B capsule is not immunogenic, and outer membrane protein vaccines provide only short-lived protection, in older children, that is restricted to serotypes in the vaccine. Protection is mediated by bactericidal antibodies that are induced by asymptomatic colonization by organisms that share lipooligosaccharides (LOS). By adolescence most children have LOS IgG that are bactericidal for most meningococcal strains. These antibodies prevent meningococcal disease, regardless of protein serotype. LOS are immunogenic at birth, and LOS antibodies can be induced by vaccination. During disease, infants make bactericidal antibodies against their infecting strains. These antibodies bind to a conserved LOS structure that is expressed well by 126E (L1,8), other L1 strains, and by L3,7 and L4,6 strains. This structure has not been definitively identified. LOS antibodies induced during infancy can prevent the monoclonal antibody (mAb), D6A, from binding to 126E LOS, and we have used this mAb, which binds to meningococci of all groups and types, as a surrogate for the LOS structure. MAb D6A binds the deeply truncated LOS of deltagalE mutants that have only a conserved basal structure, but it is not clear that this truncated LOS is the optimal immunogen. We now propose to affinity purify human IgG that binds the conserved LOS structure by 1) passage of IVIG over deltagalE LOS, and 2) by sequential passage of IVIG over L1, L1,8, L3,7 and L4,6 LOS, each coupled to Sepharose. We will assess how well each IgG kills 34 consecutive and unique endemic meningococcal case strains, and whether they can opsonize these meningococci for PMN killing. We will compare the binding of this pauciclonal IgG (pclgG) to that of mAb D6A and use Mass Spectrometry combined with chemical and enzymatic degradations to confirm the LOS structure recognized by mAb D6A and pclgG. We will immunize transgenic mice that have human immmunoglobulin loci (XenoMouse) with a deltagalE mutant to insure that this LOS structure is immunogenic for the human immune system, and characterize the functional activity of the induced IgG, as for the IVIG IgG. We will use enrichment of a coliphage display library with mAb D6A to identify a peptide mimic of the mAb D6A LOS antigen, use this peptide to immunize XenoMouse mice and functionally charaterize the induced IgG. We also will try to identify a peptide mimic that binds the human pclgG. The resulting data should confirm the suitability of a conserved LOS structure as a vaccine for the prevention of endemic group B disease in infants and young children. Website: http://crisp.cit.nih.gov/crisp/Crisp_Query.Generate_Screen

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Project Title: AMBISOME VS APHOTERICIN B IN AIDS PATIENTS WITH CRYPTOCOCCAL MENINGITIS Principal Investigator & Institution: Javaly, Kedernath; New York University School of Medicine 550 1St Ave New York, NY 10016 Timing: Fiscal Year 2001 Summary: This abstract is not available. Website: http://crisp.cit.nih.gov/crisp/Crisp_Query.Generate_Screen

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Project Title: ANALYSIS OF M. CATARRHALIS LOS: ROLE IN IMMUNITY Principal Investigator & Institution: Campagnari, Anthony A. Professor; State University of New York at Buffalo 402 Crofts Hall Buffalo, NY 14260 Timing: Fiscal Year 2001; Project Start 01-MAR-2001; Project End 28-FEB-2006 Summary: Moraxella catarrhalis, a human mucosal pathogen, is a prominent cause of otitis media in young children and lower respiratory tract infections in adults with COPD. The significant financial burden on the health care system in this country, has stimulated research studies aimed at identifying possible vaccine components expressed on the bacterial surface. Recent studies have focused on components of the bacterial outer membrane, as these structures would most likely be available for interaction with the host immune response. However, it is clear that little is known about the virulence factors and the host immune response to M. catarrhalis. One prominent bacterial surface component, implicated as a potential virulence factor, is the lipooligosaccharide (LOS). Structural studies have shown that this major glycolipid is relatively conserved among clinical isolates obtained from adults. There have been three LOS serotypes reported using polyclonal rabbit sera for detection. In addition, a comparison of M. catarrhalis LOS has shown that this structure has oligosaccharide epitopes which share homology with the LOS of 9other important Gram-negative human pathogens, including Neisseria meningitis, Neisseria gonorrhea and Haemophilus influenzae. The LOPS epitopes shared by M. catarrhalis and these other important pathogens have been implicated as potential virulence factors involved in various mechanisms of pathogenesis including adherence and invasion of mucosal cells, serum resistance and resistance to opsonophagocytosis. In addition, recent studies have shown that antibodies to M. catarrhalis LOS elicit bactericidal activity, suggesting that this molecule may be an important component of a multifactorial vaccine. Despite these data, there is very little known about the role of M. catarrhalis LOS in colonization of infection, and there are no studies reported which characterize the genes and gene products of this important bacterial component. In this proposal, we will clone and sequence genes involved in the biosynthesis and assembly of LOS and we will construct specific isogenic mutants in these genes. These LOS mutants will be evaluated in various biologic assays to begin to understand the role of this molecule in pathogenesis. Also, we will perform a detailed analysis of the human antibody response to LOS in both children and adults which will provide insight into the host-pathogen relationship. The data obtained from these studies will provide critical information to our understanding of the steps involved in the pathogenesis of M. catarrhalis infections which will lead to new insight into strategies designed to prevent disease. Website: http://crisp.cit.nih.gov/crisp/Crisp_Query.Generate_Screen

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Project Title: ANTIGEN SPECIFIC T CELL ACCUMULATION IN THE CNS Principal Investigator & Institution: Fabry, Zsuzsanna; Professor; Pathology and Lab Medicine; University of Wisconsin Madison 750 University Ave Madison, WI 53706 Timing: Fiscal Year 2001; Project Start 01-FEB-2000; Project End 31-JAN-2004 Summary: The influx of T lymphocytes into the Central Nervous System (CNS) is an important part of the pathology of various autoimmune and infectious diseases, which are associated with inflammation in the brain, including viral encephalitis, chronic meningitis and Multiple Sclerosis (MS). Typically, inflammatory reactions involve the following phases: 1.) the initial entry phase of activated T cells; 2.) the preferential accumulation and site-specific survival of a small number of activated, antigen specific T cells; 3.) recruitment phase, in which, inflammatory mediators produced by these

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antigen specific T cells contribute to the influx of non-antigen specific leukocytes resulting in an amplification of the inflammatory response. The primary goal of this application is to achieve progress toward understanding the mechanism of entry and accumulation of antigen specific T cells in the early initiation phase of CNS inflammatory reactions. The first part of our work (Specific Aims 1 and 2) will focus on understanding how antigen specific T cell responses are initiated by systemic or CNS introduction of antigens. We will compare different routes of antigenic stimulation and determine which one results in the most exacerbated T cell response in the brain parenchyma and Cerebral Spinal Fluid (CSF). Because of the difficulties in monitoring the very small numbers of antigen specific T cells found in a normal animal, we will use a novel technology already available in our laboratory, involving T cell receptor transgenic animals and their respective antigens. In this way we will be able to follow the accumulation of a monospecific T cell population in the CNS. The goal of Specific Aim 3 will be to interfere with the initial entry (phase 1) of antigen specific and nonspecific T cells into the CNS. Molecules governing this initial entry phase of T cells will be defined. Characterization of this process will be crucial to understanding the pathogenesis of disease, and developing effective immunotherapies. We believe that the successful completion of this research project will lead to an improved understanding of the role of T cells in CNS inflammatory diseases and will provide the foundation for new therapeutic methodologies for controlling CNS inflammatory diseases, such as MS. Website: http://crisp.cit.nih.gov/crisp/Crisp_Query.Generate_Screen •

Project Title: BACTERIAL PHOSPHORYLCHOLINE AND PATHOGENESIS Principal Investigator & Institution: Weiser, Jeffrey N. Associate Professor; Microbiology; University of Pennsylvania 3451 Walnut Street Philadelphia, PA 19104 Timing: Fiscal Year 2001; Project Start 01-AUG-1999; Project End 31-JUL-2004 Summary: Description (Adapted from Applicant's Abstract): The bacterial cell surface is generally considered to be highly divergent from species to species. An exception to this rule is the expression of phosphorylcholine (ChoP). This unusual prokaryotic structure is now known to be exposed on the surface of the most common pathogens infecting the human respiratory tract; Haemophilus influenzae, mycoplasma, and Streptococcus pneumoniae. In addition, based on cross-reactivity to a MAb recognizing this structure, ChoP may be present on diverse phase-variable structures on N. meningitidis, N. gonorrhoeae, P. aeruginosa, and A. actinomycetemcomitans. We have defined the genetic basis of ChoP expression and the molecular mechanism controlling its phase variation in H. influenzae. This has allowed direct genetic analysis of clinical samples to show that the ChoP+ phase variants predominate on the mucosal surface of humans. The structure, however, is the target of innate immunity mediated by binding of C-reactive protein (CRP), which is bactericidal in the presence of complement. The focus of this proposal is to define the biological role of variants both with and without ChoP using H. influenzae as a prototype human respiratory tract pathogen. In Aim 1, we will determine whether switching to the ChoP- phenotype is required in natural H. influenzae infection (otitis media, pneumonia, bacteremia, and meningitis) to evade clearance by CRP and bactericidal anti-ChoP IgG. The ChoP phenotype in vivo will be determined by direct genetic analysis and compared to the local concentration of CRP and anti-ChoP antibody during infection. The local expression and concentration of CRP in the upper respiratory tract will be investigated. In Aim 2, we will determine how ChoP contributes to persistence on the mucosal surface. Genetically defined H. influenzae mutants with constitutive ChoP-on and ChoP-off phenotypes will be used to determine whether this host membrane-like

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structure contributes to (a) resistance to respiratory tract antibacterial peptides including LL-37 and tracheal antimicrobial peptide (TAP), and (b) colonization by functioning as a bacterial adhesin to host epithelial cells via putative ChoP ligands including GalNAcb 14Gal on the asialo-GM1 glycolipid and the platelet activating factor receptor. The blocking of complement mediated killing by naturally acquired secretory IgA recognizing ChoP will be explored as an explanation for the selection of the ChoP+ phenotype on the mucosal surface, despite the increased susceptibility of this phenotype to CRP and complement. Website: http://crisp.cit.nih.gov/crisp/Crisp_Query.Generate_Screen •

Project Title: BACTERIAL VACCINE ANTIGEN DISCOVERY Principal Investigator & Institution: Paoletti, Lawrence C.; Brigham and Women's Hospital 75 Francis Street Boston, MA 02115 Timing: Fiscal Year 2003; Project Start 30-SEP-2003; Project End 31-AUG-2004 Summary: (provided by applicant): New directions in bacterial vaccine discovery may arise from studies of host-microbe interactions, particularly through the use of a newly described technology: the dynamic in vitro attachment and invasion system (DIVAS). DIVAS was developed to study bacterial attachment and invasion with cells held at specific and controlled conditions of growth, metabolism, and nutrient levels. Results from experiments performed with DIVAS and group B Streptococcus (GBS) type III strains substantiated earlier findings that capsular polysaccharide is not critical for invasion of respiratory epithelial cells. Moreover, GBS invaded these cells only when held at a fast as opposed to a relatively slower rate of growth and they expressed several proteins solely under growth conditions conducive for invasion. In this proposal, we seek to test the hypothesis that GBS proteins involved with invasion of eukaryotic cells are new and important targets of protective immunity. GBS is a major cause of neonatal sepsis and meningitis, and is increasingly prevalent among nonpregnant adults and the elderly with underlying illnesses. Preclinical and clinical trials have been successfully performed with protein conjugate vaccines prepared with polysaccharides from most of the nine currently known GBS serotypes. GBS protein antigens with virulence properties have been described and some with vaccine potential have been tested preclinically. In this proposal, we seek to use DIVAS to identify physiological conditions conducive for bacterial attachment/invasion of eukaryotic cells using GBS as a model pathogen. We plan to isolate and identify GBS membrane proteins expressed solely under invasive conditions (Specific Aim 1). Several of these newly expressed proteins will be purified directly from GBS, or cloned and recombinantly expressed, and tested as vaccine candidates in mice (Specific Aim 2). Findings from these studies utilizing this unique approach to vaccine antigen discovery could be directly applied to other bacterial pathogens including those on the category A bioterrorism list. Website: http://crisp.cit.nih.gov/crisp/Crisp_Query.Generate_Screen

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Project Title: MENINGITIS

BIOACTIVITIES

OF

PNEUMOCOCCAL

CELL

WALL

IN

Principal Investigator & Institution: Tuomanen, Elaine I. Chair, Professor; St. Jude Children's Research Hospital Memphis, TN 381052794 Timing: Fiscal Year 2001; Project Start 01-JUN-1989; Project End 31-JAN-2005 Summary: The pneumococcus remains the cause of meningitis with the greatest morbidity and mortality in children and older adults. This pattern persists despite the use of antibiotics of exceptionally rapid bactericidal activity. Over the past 10 years of

Studies 11

this proposal we have sought to understand the biochemical basis of the inflammatory response to pneumococci in the subarachnoid space. We established that the pneumococcal cell wall is a library of inflammatory components which incites the cytokine, coagulation and arachidonate cascades and directly injures endothelial cells of the blood brain barrier. Further, we established that the release of cell wall during antibiotic-induced death engenders a dramatic host response that is responsible for serious injury to host tissues. This provided a rationale for use of agents like dexamethasone that can act as partner drugs with antibiotics to selectively control the injurious components of the host defense response. The current proposal seeks to determine the molecular details of the mechanism of pneumococcal invasion into brain and how neuronal cells are killed during meningitis. Blocking information decreases some sequelae of infection but does not appear to be sufficient in controlling neuronal loss, particularly for pneumococcal disease. Over half of the current survivors of this injection still have major permanent sequelae. Understanding this process will allow design of agents to specifically attenuate these ongoing losses. We propose to apply our expertise in the identification and characterization of pneumococcal surface components, to map the process of transcytosis across the blood brain barrier. We will identify the pneumococcal components involved, specifically focusing on CbpA. This protein is required for pneumococcal invasion. Secondly, we will characterize the process of pneumococcal translocation in terms of the intracellular vesicle and the signaling process. Involvement of the PAF receptor that binds pneumococci and sIgA that ligates CbpA in actual translocation will be determined. Finally, we will investigate preliminary evidence that upon inhibition of apoptosis suggest this is an important contributor to sequelae. The detailed mechanism appears to be novel and will potentially instruct cell biology as well as pathogenesis. Website: http://crisp.cit.nih.gov/crisp/Crisp_Query.Generate_Screen •

Project Title: BIOLOGY OF H INFLUENZAE HIA AND HSF ADHESINS Principal Investigator & Institution: St Geme, Joseph W. Associate Professor; Pediatrics; Washington University Lindell and Skinker Blvd St. Louis, MO 63130 Timing: Fiscal Year 2001; Project Start 15-DEC-1998; Project End 30-NOV-2003 Summary: Haemophilus influenzae is common cause of localized respiratory tract disease, including otitis media, sinusitis, bronchitis, and pneumonia. Less commonly, this organism causes serious systemic disease, such as meningitis, endocarditis, and septicemia. The initial step in the pathogenesis of H. influenzae disease involves colonization of the upper respiratory mucosa. We have identified a high-molecularweight protein called Hia, which is present in nontypable (nonencapsulated) H. influenzae and promotes attachment to human epithelium. In addition, we have identified a homolog of Hia called Hsf, which is univerally present among encapsulated H. influenzae and also mediates in vitro adherence. Interestingly, Hia has a predicted molecular mass of approximately 114 kDa and is not detectable by coventional transmission electron microscopy, while Hsf has a predicted molecular mass of approximately 245 kDa and is associated with the presence of short, thin surface fibrils visible by negative staining electron microscopy. Based on our in vitro results, we speculate that Hia and Hsf are important colonization factors. In the present proposal, we plan to characterize the pathway by which Hia and Hsf are localized on the surface of the organism. In particular, we will define the structural features of these proteins that direct them to the periplasm and facilitate their translocation across the outer membrane. We will dissect the influence of an unusual N-terminal extremity, a Cterminal domain predicted to form a beta-barrel, and a putative ATP-binding motif. In

12 Meningitis

additional studies, we will examine the architecture of Hia and will investigate the relationship between structure and adhesive activity, focusing in particular on the role of a predicted coiled coil motif. We will also determine whether Hia and Hsf function interchangeably in an encapsulated strain. From a practical perspective, the results of these experiments may facilitate efforts to develop a vaccine protective against non-type b H. influenzae and suggest targets for novel antimicrobials with activity against a broad range of gram-negative bacteria. More generally, they may provide fundamental insights into the biogenesis of non-pilus adhesins and the nature of the host-microbial relationship. Website: http://crisp.cit.nih.gov/crisp/Crisp_Query.Generate_Screen •

Project Title: BIOMARKER OF NEUROPROTECTANT EFFICACY IN ALS Principal Investigator & Institution: Gabbita, Somasundar P.; Phase 2 Discovery, Inc. 3130 Highland Ave, 3Rd Fl Cincinnati, OH 45219 Timing: Fiscal Year 2003; Project Start 15-APR-2003; Project End 31-MAR-2004 Summary: (provided by applicant): The objective of this Phase I feasibility study is to develop a reliable biomarker of neuronal damage in a validated mouse model of amyotrophic lateral sclerosis (ALS). Currently, there is no widely accepted biomarker to quantify ALS-induced motor neuron injury. Previously, we have shown that neuronal degeneration in rodents and humans results in cleavage of the cytoskeletal protein MAP-tau. Our laboratory has developed a sensitive ELISA that specifically measures cleaved MAP-tau (C-tau) in rodent models of traumatic brain injury and bacterial meningitis. Using this C-tau ELISA assay, our preliminary studies demonstrate that spinal cord C-tau levels are tenfold higher in late stage symptomatic ALS (G93A-SOD1 mutation) mice as compared to control mice. Using a rotorod motor performance assay, Hensley et al. have recently demonstrated that administration of Nor-dihydroguaiaretic acid (NDGA) to ALS mice delays onset of neurologic deficits. We hypothesize that 1) Ctau is a reliable biomarker of motor neuron injury in ALS mice and 2) C-tau can serve as a screening tool to identify neuroprotectant drags for treating ALS. We will test these hypotheses by determining the relationship between C-tau levels and progression of neurologic deficits in ALS mice. Furthermore, we will test whether the demonstrated neuroprotectant effect of NDGA that delays onset of neurologic deficits in ALS mice also exerts expected effects on C-tau levels. Our Specific Aims are: Specific Aim 1: Compare spinal cord C-tau levels in ALS mice and control mice at 120 days. Specific Aim 2: Determine the relationship between C-tau levels and neurologic deficits in ALS mice as compared to controls. Neurologic deficits and spinal cord C-tau levels will be determined in ALS and age-matched control mice and statistically compared from the presymptomatic stages to late symptomatic stages of disease progression. Specific Aim 3: Determine if C-tau levels reliably quantify the effect of a demonstrated neuroprotectant drag intervention in ALS mice. C-tau levels and rotorod performance will be determined and compared as a function of NDGA treatment. Website: http://crisp.cit.nih.gov/crisp/Crisp_Query.Generate_Screen

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Project Title: BIOMARKER OF NEUROTOXICITY IN MENINGITIS Principal Investigator & Institution: Mulchahey, James J. Assistant Professor; Phase 2 Discovery, Inc. 3130 Highland Ave, 3Rd Fl Cincinnati, OH 45219 Timing: Fiscal Year 2002; Project Start 01-AUG-2002; Project End 31-JUL-2003 Summary: (provided by applicant): The objective of this Phase I application is to develop a sensitive biomarker for quantifying neurotoxicity and neuroprotectant

Studies 13

efficacy in meningitis. Previous research documents that brain injury caused by meningitis affects multiple brain areas with a heterogeneous distribution. We have previously shown that the cytoskeletal protein MAP-tau is cleavedin brain during axonal degeneration. We developed a sensitive ELISA that specifically quantifies this biomarker of neuronal degeneration, cleaved-tau (C-tau). Our preliminary studies demonstrate that levels of C-tau are increased over 300-fold in an animal model of group B streptococcus meningitis (GBM). We will use the well-documented neurotoxicity of GBM to validate the C-tau as a biomarker of neurotoxicity and a measure of neuroprotectant efficacy. Our Specific Aims are: Specific Aim 1: Determine whether GBM results in a time-dependent elevation in brain, plasma and CSF concentrations of C-tau. Specific Aim 2: Determine it C-tau levels in brain, CSF and plasma correlate with traditional measures of neuronal damage in GBM. Specific Aim 3: Determine whether perihperal tissues such as liver, kidney or spleen are potential sources of C-tau measured in GBM. Specific Aim 4: Determine whether a neuroprotectant intervention known to be effective in GBM has predictable effects on Ctau levels. PROPOSED COMMERCIAL APPLICATIONS: The C-tau ELISA may be used to quantify the severity of brain injury in meningitis and to quantify the effects of neuroprotectant interventions in basic science, preclinical and clinical research settings. Website: http://crisp.cit.nih.gov/crisp/Crisp_Query.Generate_Screen •

Project Title: CLINICAL TRIALS WITH SOLID INTRACRANIAL TUMORS & NEOPLASTIC MENINGITIS Principal Investigator & Institution: Bigner, Darell D. Jones Cancer Research Professor; Duke University Durham, NC 27706 Timing: Fiscal Year 2002 Summary: Given the toxicity and poor survival results after current treatment modalities, regional therapy using monoclonal antibody-based strategies and alkylators may have clinical potential. The PI proposes to define the MTD and therapeutic benefits of 9 different approaches, 6 for solid intracranial tumors and 3 for neoplastic meningitis. For solid tumors, 4 approaches involved intratumoral or intracystic infusions: (1) 131-I81C6, (2) 211-Att-ch81C6, (3) ch81C6-streptavidin followed by 211-At-biotin or 131-Ibiotin, (4) 211-At or I-131 conjugated alpha-EGFRvIII/ fragments; and 2 approaches only intratumoral infusions: (1) MRI (a-EGFRvIII-scFv-pseudomonas toxin), and (2) TP38 (TGFa-pseudomonas toxin). For neoplastic meningitis, two treatment studies were chemotherapy based: melphalan and camptothecin analogues and the rest utilized 131-IMel-14, 131-I-81C6, and other anti-medullobastoma antibodies and genetic variants. Website: http://crisp.cit.nih.gov/crisp/Crisp_Query.Generate_Screen

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Project Title: ANTIGENICITY

COMPUTATIONAL

ANALYSIS

OF

CARBOHYDRATE

Principal Investigator & Institution: Woods, Robert J. None; University of Georgia 617 Boyd, Gsrc Athens, GA 306027411 Timing: Fiscal Year 2003; Project Start 01-MAR-1997; Project End 30-APR-2007 Summary: (provided by applicant): Group Beta Streptococcus and Neisseria meningitidis are leading causes of neonatal sepsis and meningitis. The increasing use of carbohydrate-based conjugate vaccines is founded on the observation that antibodies against the type-specific bacterial capsular polysaccharides (CPS) are often protective, and-is driven by the increasing prevalence of antibiotic resistant strains. However, the relationships between the carbohydrate sequence in the CPS and antigenicity are poorly

14 Meningitis

understood. Similarly, the immune response to the CPS is structure-sensitive; some are poor immunogens and others good. The goal of this proposal is to provide an understanding of the structural features of antigenic oligosaccharides that are responsible for mediating the affinity and specificity of their interactions with antibodies. Ultimately, this information would form a basis for the rational development of more effective antibacterial vaccines. We have selected three systems (Aims 1-3) for study that display complementary levels of complexity. In Aims 1 and 2, we will use computational and experimental methods to determine the conformational properties of the bacterial CPSs from iV. meningitides and Group B Streptococcus, both free and bound to monoclonal antibody fragments. This information will provide a structural basis for interpreting the antigenicities and antibody specificities for these systems, as well as assist in the determination of the conformation of the immunodominant regions. To aid in the development and validation of the computational methods, in Aim 3, we will examine the properties of the related anionic sugars in glycosaminoglycans (GAGs), for which considerable experimental data exist. Computational methods, such as molecular dynamics (MD) simulations, are useful aids in the conformationat analysis of oligosaccharides and oligosaccharide-protein complexes; yet at present they have been developed only for neutral carbohydrates. The CPSs from N. meningitides and Group B Streptococcus, and from many other pathogenic bacteria, contain anionic carbohydrate residues. Accurate modeling of these molecules will require the extension and validation of our existing carbohydrate force field parameters (GLYCAM). To assist in this development, we will examine a number of anionic GAGs, such as hyaluronan, chondroitin and heparin sulfate as well as heparin, free and complexed to antithrombin III, whose conformational properties have been well characterized experimentally. Validation of the MD simulations will be based on comparisons with NMR and X-ray data and binding affinity measurements. Website: http://crisp.cit.nih.gov/crisp/Crisp_Query.Generate_Screen •

Project Title: CONGENIC NEOFORMANS

STRAINS

OF

SEROTYPE

A

CRYPTOCOCCUS

Principal Investigator & Institution: Wickes, Brian L. Assistant Professor; Microbiology and Immunology; University of Texas Hlth Sci Ctr San Ant 7703 Floyd Curl Dr San Antonio, TX 78229 Timing: Fiscal Year 2003; Project Start 01-FEB-2003; Project End 31-JAN-2006 Summary: (provided by applicant): Cryptococcus neoformans is an important human fungal pathogen that can cause life-threatening infections. Infections are opportunistic in nature and typically occur in immunocompromised patients, with AIDS patients being at greatest risk. Cryptococcosis is usually manifested as meningitis, which is often treated as incurable in AIDS patients and can require life-long antifungal therapy. Infections are worldwide with approximately 90- 95% of all infections being caused by serotype A strains. The importance of this fungus as a pathogen resulted in the solicitation and funding of a genome-sequencing project. The strain that was chosen, however, was a serotype D strain, which causes less than 10% of all infections. This strain was chosen because sexually compatible strains (designated MATalpha and MATa) exist, whereas compatible serotype A strains are not available due to the absence of a fertile MATa strain. An added significance of mating in C. neoformans is that virtually all infections are caused by one mating type (MATalpha). We have recently identified a serotype A MATa strain that is fertile and crosses efficiently with serotype A MATalpha strains, including strain H99, the standard laboratory strain for this serotype. The existence of this strain provides a powerful tool for studying C. neoformans because

Studies 15

it will enable traditional genetic studies to be performed in serotype A strains. This proposal consists of three aims. The first aim will prepare two congenic pairs of strains that differ only in mating type. Genetic markers will be inserted into these strains to make them useful tools for future genetic and molecular studies. The second aim will test the role of mating type in virulence. By using congenic strains in these experiments, the role of information at the mating type locus in virulence can be tested independent of non mating-type information. In the last aim, the serotype A MATa mating type locus will be cloned and mapped to determine its size. Presently three out of the four mating type loci of C. neoformans var. neoformans (comprised of serotype A and D) are in various stages of characterization. Isolation of the serotype A MATa locus will complete this study, which is significant because of the role that mating type plays in virulence. Website: http://crisp.cit.nih.gov/crisp/Crisp_Query.Generate_Screen •

Project Title: CORE--PHASE I / II CLINICAL TRIALS CORE Principal Investigator & Institution: Reardon, David; Duke University Durham, NC 27706 Timing: Fiscal Year 2002; Project Start 30-SEP-2002; Project End 31-AUG-2004 Summary: (provided by applicant): Core 3 will design, implement and review all clinical trials sponsored by the SPORE application. There are four major areas of therapeutic intervention to be evaluated by clinical trials overseen by the core. First, therapeutics to overcome chemoresistance will be evaluated. UG-BG, a potent inhibitor of 06alkylguanine- DNA alkyltransferase (AGT), the major mediator of chemoresistance among malignant gliomas, is ineffective and associated with significant toxicity when administered systemically. Therefore we will evaluate the compartmental administration of Ub-BG, as well as other agents identified in Project 4, via intrathecal injection for patients with neoplastic meningitis (NM) and intratumorally via intracerebral microinfusion (ICM) for patients with malignant brain tumors. Second, we will continue to evaluate therapeutics targeting tumor- specific markers (Project l), including armed and unarmed monoclonal antibodies (MAbs) and recombinant immunotoxins, delivered compartmentally via injection into the resection cavity or into the intrathecal space, or via direct intratumoral microinfusion. Therapeutic targets for these agents include EGFR, EGFRvIII, tenascin C, GP 240, MDRP3, GPNMB as well as others identified in Project 2. Our recent Phase I1 study showed that I-labeled antitenascin MAb 81C6 significantly improved survival for patients with newly diagnosed malignant glioma compared to conventional external beam radiotherapy and chemotherapy. Therefore, we are advancing our extensive preclinical and clinical studies with "'1-81 C6 and investigating the role of human and mouse chimeric MAbs, MAb fragments such as F(ab'), and alternative radioisotopes such as astatine- 211. The third major area of focus is the evaluation of administering therapeutics by intracerebral microinfusion as a method of achieving maximum intratumoral concentration while minimizing systemic exposure. and thereby significantly improving drug therapeutic index. Fourth, we are continuing to evaluate cytotoxic chemotherapeutics that have shown promising, preclinical efficacy in the extensive Duke panel of brain tumor cell lines and xenograft models. Website: http://crisp.cit.nih.gov/crisp/Crisp_Query.Generate_Screen

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Project Title: CORE--X RAY CRYSTALLOGRAPHY Principal Investigator & Institution: Sundaramoorthy, Munirathinam; University of Kansas Medical Center Msn 1039 Kansas City, KS 66160

16 Meningitis

Timing: Fiscal Year 2001; Project Start 30-SEP-2001; Project End 31-AUG-2006 Summary: The success of several sequence genomics projects heralds a new era in the study of the gene products and many of them are attractive targets for drug design. The new proteomics research uses multidisciplinary approach to study the "cellular protein universe" using several high-throughput methods. X-ray crystallography plays a pivotal role in both proteomics and structure based drug design. The University of Kansas Medical Center has envisioned a plan to establish "Kansas City Proteomics Initiative" and will set up an X-ray crystallography lab as a part of this plan in the Department of Biochemistry. This will serve as the Core facility for COBRE investigators who apply crystallography to their research. Funds are requested from COBRE for its maintenance. The Principal Investigator will direct the Core and his Research Project is summarized below. Gram-negative bacteria Neisseria meningitidis and Escherichia coli K1 are associated with sepsis and meningitis. There is no promising vaccine against N. Meningitidis group B and E. coli K1. Their capsular polysaccharides are sialylated, which are structurally similar to those found in mammalian glycolipids. This host mimicry plays and important role in the pathogenecity by allowing the bacteria to evade the host's immune system. Thus, sialic acid (N-acetylneuraminic acid, NeuNAc) is a virulence factor in these pathogenic bacteria. The biosynthesis of polysialic acid involves the formation and activation of NeuNAc. In bacteria, these two steps are catalyzed by NeuNAc synthase and CMP-NeuNAc synthetase, respectively. Our long-term goal is to study the structure-function relations of these enzymes using molecular biology, protein chemistry, and crystallography methods and use the structural knowledge of rational drug design. The specific aims of this application are: Aim 1: NeuNAc synthases from N. meningitidis and E. coli K1. In bacteria, sialic acid is synthesized by the condensation of N-acetylmannosamine with polyenolpyruvate by NeuNAc synthase. The human enzyme uses the phosphorylated substrates. The differences in the substrate specificity between human and bacterial enzymes may be due to different active site structures. Selective inhibition of bacterial enzyme is a viable strategy to prevent the synthesis of sialic acid in the pathogen. We will overexpress, purify, and crystallize NeuNAc synthases from N. meningitidis and E. coli K1 for structural-functions studies. Aim 2: CMP-NeuNAc synthetase from E. coli K1. The E coli K1 CMP-NeuNAc synthetase is twice as large as the other bacterial enzymes and about the same size as the mouse enzyme, the only mammalian CMP-NeuAc synthetase is sequenced. The N-terminal half of the E. coli K1 sequence is highly homologous to other bacterial sequences, but the full-length sequence share a limited homology with the mouse sequence. We will crystallize the E. coli K1 CMP-NeuAc synthetase for structure determination. The structural differences of these functional homologues will be used to design drugs to target the bacterial enzymes specifically. Website: http://crisp.cit.nih.gov/crisp/Crisp_Query.Generate_Screen •

Project Title: COXSACKIE MYOCARDITIS AND VIRAL PERSISTENCE IN THE HEART Principal Investigator & Institution: Whitton, J Lindsay. Professor; Scripps Research Institute 10550 N Torrey Pines Rd La Jolla, CA 920371000 Timing: Fiscal Year 2003; Project Start 01-JAN-1998; Project End 31-DEC-2007 Summary: (provided by applicant): Coxsackieviruses (CVB) are important human pathogens, causing myocarditis, meningitis, and other diseases, which may be lethal, especially in neonates. This proposal has five specific aims focused on CVB pathogenesis (Aims 1-3) and immunity (Aims 4, 5). Aim 1. Does CVB preferentially infect proliferating cells in vivo? We have shown, in tissue culture, that the cell cycle exerts a

Studies 17

dramatic effect on CVB gene expression & virus production. Is this also true in vivo, in the heart, central nervous system, or immune system? We shall identify proliferating cells in these tissues, and will determine if they are more susceptible to infection. If the heart is injured in vivo, does this alter its susceptibility to subsequent CVB infection? Aim 2. What component of the virus responds to the host cell cycle? Next, we shall study the viral side of the equation. We hypothesize that the internal ribosome entry site may be the viral "response element"; this will be evaluated in tissue culture, and in vivo. Aim 3. What role do the proposed CVB receptors play in tropism & pathogenesis in the heart & CNS? We shall evaluate the expression levels of the receptors. Do they change with age? Following infection / tissue damage, are the receptors upregulated in neighboring cells? Aim 4. How do CD8+ T cells contribute to the control of CVB infection? We have shown that, during CVB infection, CD8+ T cells can reduce viral titers, and that this effect does not require perforin. What effector functions are involved in the antiviral effect? Do these effector functions contribute to tissue damage?Aim 5. How does CVB so effectively avoid inducing strong CD8+ T cell responses? Most virus infections induce high levels of antiviral CD8+ T cells. How does CVB avoid this? Can we rectify the situation? Website: http://crisp.cit.nih.gov/crisp/Crisp_Query.Generate_Screen •

Project Title: DEAFNESS AND OSSIFICATION IN LABYRINTHITIS OSSIFICANS Principal Investigator & Institution: Brodie, Hilary A. Otolaryngology; University of California Davis Sponsored Programs, 118 Everson Hall Davis, CA 95616 Timing: Fiscal Year 2001; Project Start 01-JAN-2000; Project End 31-DEC-2004 Summary: (Adapted from the Investigator's Abstract) Labyrinthitis ossificans (LO) is the growth of pathologic new bone within the lumen of the cochlea. It is multifactorial in origin and may result in deafness. Profound hearing loss and LO in children are most commonly associated with meningogenic labyrinthitis. The relationship of LO to meningogenic cochlear pathology and its mechanism of induction have not been clearly defined. Cochlear implants are a significant treatment option for improving hearing and quality of life in these patients. However, LO can reduce the efficacy of cochlear implantation. The long-term objective of this research program is to understand the mechanisms which lead to the development, progression, and destructive aspects of LO. Such an understanding may lead to new strategies to prevent the devastating effects of hearing loss associated with this disease. The specific aims of this application are: (1) to correlate hearing loss with the temporal and spatial progression of bacterial meningitis from the subarachnoid space to the cochlea; (2) to correlate hearing loss with the temporal and spatial sequence for both labyrinthine fibrosis and ossification and the histopathology of cochlear tissues: spiral ganglion, organ of Corti, Reissner's membrane, stria vascularis, and spiral ligament; (3) to determine the relationship of bone lining cells to osteoblast formation and recruitment during labyrinthine neo-ossification; and (4) to determine the effects of decomplementation, non-steroidal anti-inflammatory compounds, and bacteriostatic vs. bactericidal antibiotics on neo-ossification and hearing loss. There are four hypotheses/specific aims outlined and they are as follows: Bacterial invasion of the cochlear labyrinth from the subarachnoid space correlates with hearing loss and occurs principally via the cochlear aqueduct and not the internal auditory canal; The destruction of cochlear tissue occurs subsequent to the arrival of inflammatory cells and not with the appearance of bacteria within the cochlea. Hearing loss may occur prior to observable pathology and reflect central auditory damage related to meningitis; Bone lining cells of the endosteum are activated and become mature osteoblasts and are the principal source of neo-ossification in labyrinthitis

18 Meningitis

ossificans; and The inflammatory response to suppurative labyrinthitis includes fibrosis and neo-ossification formation and causes cochlear tissue destruction. Inhibition of this process will result in a reduction in both hearing loss and bone deposition. Methods used: The investigators propose to use an experimental gerbilline model of LO, histomorphometry, fluorescent bone histomorphometry, transmission and scanning electron microscopy, autoradiography, and auditory brainstem evoked response. Website: http://crisp.cit.nih.gov/crisp/Crisp_Query.Generate_Screen •

Project Title: DETERMINANTS OF INNATE IMMUNITY TO GROUP B STREPTOCOCCI Principal Investigator & Institution: Levy, Ofer; Brigham and Women's Hospital 75 Francis Street Boston, MA 02115 Timing: Fiscal Year 2002; Project Start 01-MAR-2002; Project End 28-FEB-2006 Summary: (provided by applicant): The central goal of this project is to understand the role of innate immunity in infections caused by Group B streptococci (S. agalactiae or GBS). GBS are the major cause of neonatal sepsis and meningitis and thus a leading cause of neonatal morbidity and mortality. GBS-induced inflammatory mediators include tumor necrosis factor (TNF) that, when excessive, can contribute to host morbidity and mortality. Recent evidence suggests that a variety of Gram-positive bacterial surface molecules activate the innate immune system via phagocyte innate immune receptors including cellular differentiation antigen-14 (CD14), complement receptors-3 and -4 (CR3/4), and Toll-like receptor 2 (TLR2). The candidate will further characterize bacterial and host determinants of innate immunity to GBS and test the following hypotheses: Specific molecular interactions between GBS surface components and phagocyte innate immune receptors mediate the host inflammatory response to GBS infection, that such responses are down-regulated by neutrophil-derived antimicrobial peptides that bind and neutralize inflammatory GBS surface molecules, and that these pro- and anti-inflammatory innate immune responses differ between newborns and adults. In Aim 1, GBS surface components that activate host phagocytes (i.e., neutrophils and monocytes) will be identified and characterized. In Aim 2, newborn and adult phagocytes will be compared with respect to expression and function of CR3/4, CD14, and TLR2. In Aim 3, putative neutrophil-derived peptides with anti-inflammatory activity against GBS will be isolated and characterized. The candidate seeks an intensive, formal, mentored training as preparation for becoming an independent scientist. As a specialist in pediatric infectious diseases, his long-term goal is to identify molecular pathways of innate immunity that might someday be modulated to improve outcomes of GBS and other bacterial infections in neonates. Website: http://crisp.cit.nih.gov/crisp/Crisp_Query.Generate_Screen

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Project Title: MENINGITIS

DISCONTINUATION

ANTIFUNGAL

FOR

CRYPTOCOCCAL

Principal Investigator & Institution: Aberg, Judith; University of California San Francisco 500 Parnassus Ave San Francisco, CA 94122 Timing: Fiscal Year 2001 Summary: This abstract is not available. Website: http://crisp.cit.nih.gov/crisp/Crisp_Query.Generate_Screen

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Project Title: E COLI TRANSLOCATION ACROSS INTESTINAL EPITHELIUM Principal Investigator & Institution: Pietzak, Michelle M.; Children's Hospital Los Angeles 4650 Sunset Blvd Los Angeles, CA 90027 Timing: Fiscal Year 2001; Project Start 01-JUL-1999; Project End 30-JUN-2004 Summary: This application for a Mentored Clinician Scientist Development Award (K08) seeks support for Michelle Pietzak, M.D., who has recently completed her fellowship in Pediatric Gastroenterology and Nutrition and joined the faculty as an Instructor of Pediatrics at Childrens Hospital Los Angeles (Assistant Professor pending). Under the mentorship of Kwang Sik Kim M.D., Dr. Pietzak will continue to pursue her basic investigations into the mechanisms by which Escherichia coli (E. coli) is able to translocate across intestinal epithelium and cause sepsis. E. coli is a leading cause of severe bacterial infections in premature infants, neonates, immunocompromised hosts, and children with central lines and primary intestinal diseases. Dr. Kim is a prominent researcher in the field of the pathogenesis of E. coli meningitis, having (a) established the virulence roles for the K1 capsular polysaccharide, outer membrane protein A, and S fimbriae using in vitro and in vivo models of the blood brain barrier developed in his lab and (b) identified several novel genes thought to be responsible for invasion of E. coli strain RS218 across the blood brain barrier, using these same models. E. coli strain RS218 is a clinical isolate from the cerebrospinal fluid of a human neonate with E. coli meningitis. Dr. Pietzak's project is focused on investigating the mechanisms by which E. coli strain RS218 is able to penetrate the intestinal epithelial barrier, both in vitro and in vivo. In earlier studies, Dr. Pietzak has used strain E44, a spontaneous rifampin resistant mutant of E. coli strain RS218, to demonstrate that this bacterium is able to invade two intestinal epithelial cell lines, Caco-2 and C2BBe-1. The specific aims of this proposal are to further characterize the invasive phenotype of E-44 in vitro, using both gentamicin invasion assays and a trans-well system. An in vivo model, using neonatal rats, will also be used to test the invasive phenotype of E-44. Environmental factors, which mimic the intraluminal gastrointestinal milieu, will be employed to examine their effects on E. coli invasion and translocation in vitro. The virulence of TnphoA transposon mutants, created in our lab and already shown to be noninvasive to the blood brain barrier in vitro and in vivo, will be determined for the intestinal epithelial barrier both in vitro and in vivo. The precise contribution of these genes to the intestinal translocation of E. coli strain RS218 will then be examined using additional molecular techniques. Under Dr. Kim's mentorship, and with Institutional support, Dr. Pietzak will be able to perfect her techniques in cell tissue culture and animal models of bacterial infection, as well as acquire new knowledge and skills in the sciences of bacterial genetics and molecular pathogenesis. Nurturing these ambitions in a collaborative environment will help Dr. Pietzak achieve her goal of becoming an independent investigator, who hopefully will be able to bring the results of her investigations from the benches of the lab to the bedsides of chronically ill children with gastrointestinal and other diseases. Website: http://crisp.cit.nih.gov/crisp/Crisp_Query.Generate_Screen

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Project Title: E. COLI BINDING TO BRAIN ENDOTHELIAL CELLS Principal Investigator & Institution: Kim, Kwang S. Professor and Director; Pediatrics; Johns Hopkins University 3400 N Charles St Baltimore, MD 21218 Timing: Fiscal Year 2001; Project Start 01-APR-2000; Project End 31-MAR-2004 Summary: (Adapted from the Applicant's Abstract): The principal investigator has established an infant rat model of experimental hematogenous meningitis, which mimics human E. coli meningitis (i.e., hematogenous infection of the meninges without

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the need for adjuvant or direct inoculation of bacteria into cerebrospinal fluid). They have established an in vitro model of blood-brain barrier using brain microvascular endothelial cells (BMEC). Using both in vitro and in vivo systems and the BMEC model, they have shown that successful traversal of E. coli across the blood-brain barrier may be a complex process involving separate steps of E. coli-BMEC interactions, i.e., binding to BMEC and invasion of BMEC. They have shown that S fimbriae are the major E. coli structures contributing to binding to BMEC. They therefore hypothesize that the role of S fimbriae is to have a more intimate contact for circulating E. coli to BMEC to withstand blood flow, which may be required for subsequent crossing of the blood-brain barrier, but there is no information to support this hypothesis. The overall aim of the proposal is to study the role of binding via S fimbriae in the pathogenesis of E. coli meningitis, both in vitro (using BMEC in culture) and in vivo (using an experimental animal model of hematogenous E. coli meningitis that closely mimics the pathogenesis of human E. coli meningitis). The specific aims are: (1)to continue to construct isogenic S fimbriae operon deletion (delta sfaII) mutant of invasive E. coli K1 strain RS218 by chromosomal gene replacement (allelic exchange); (2)to examine the ability of S fimbriae negative (delta sfaII) mutant to bind and invade brain microvascular endothelial cells (BMEC) in vitro and in vivo; (3)to determine whether it is possible to restore the ability of S fimbriae negative (delta sfaII) mutant to bind and invade BMEC by complementation with the cloned sfaII gene cluster; (4)to assess which specific subunits of S fimbriae are responsible for binding to BMEC by constructing single gene deletion mutants (delta sfaIIS, delta sfaIIA, delta sfaIIG, delta sfaIIH); (5)and to identify and characterize BMEC glycoprotein(s) interactive with S fimbriae. Website: http://crisp.cit.nih.gov/crisp/Crisp_Query.Generate_Screen •

Project Title: EARLY LYME--CNS INVASION Principal Investigator & Institution: Coyle, Patricia K. Associate Professor; State University New York Stony Brook Stony Brook, NY 11794 Timing: Fiscal Year 2001 Summary: Lyme disease is an important emerging infection caused by a tick borne spirochete B. burgdorfi (Bb). Originally characterized as an arthritis, it appears that neurologic involvement is also a significant early manifestation of the disease in North America. Neurologic complications of the disease can involve meningitis, facial nerve palsy (or other cranial neuropathy) and radiculoneuritis in early disseminated infection. It is also unclear at what point the infection changes from a localized skin erythema migrans (EM) to an early disseminated syndrome. The objective of this study is to characterize the frequency, clinical correlates and outcome of central nervous system infection in early Lyme disease. The study parameters will include clinical features, immune response, cerebrospinal fluid changes, neurocognitive disturbances, and psychosocial aspects of the disease. Website: http://crisp.cit.nih.gov/crisp/Crisp_Query.Generate_Screen

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Project Title: EHRLICHIA CHAFFEENSIS SURFACE PROTEINS Principal Investigator & Institution: Walker, David H. Professor; Pathology; University of Texas Medical Br Galveston 301 University Blvd Galveston, TX 77555 Timing: Fiscal Year 2001; Project Start 30-SEP-1991; Project End 31-MAR-2005 Summary: (provided by the applicant): The long-term goal of this research is the elucidation of the mechanisms of protective immunity against Ehrlichia chaffeensis, the causative agent of human monocytotropic ehrlichiosis (HME). Achievement of this goal

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requires knowledge of which humoral and cellular immune mechanisms stimulated by ehrlichiae are effective in the clearance of ehrlichiae. HME is a life-threatening tickborne infection associated with adult respiratory distress syndrome, meningitis, and shock in immunocompetent patients, overwhelming infection in immunocompromised patients, and a fatality rate of 2.7 percent. More than 2,200 cases have been diagnosed with laboratory confirmation, and the incidence is 1,000 cases per 1,000,000 population in tick-exposed rural populations. The specific aims test the hypothesis that the immunodominant, surface-exposed p28 antigens stimulate protective immunity by antibodies and cellular mechanisms and determine the importance and mechanisms) of antibodies and cellular mechanisms of protective immunity in mouse models of HME. The research design includes sequencing of the loci of the p28 multigene families of major immunodominant surface proteins of Ehrlichia muris and a related ehrlichia (IOE) DNA and recombinant protein vaccines in the E. muris mouse model and the highly pathogenic IOE-C57BL/6 mouse model. Humoral immunity will be passive polyclonal and monoclonal antibodies to the conserved and variable regions of p28 families in IOE-infected mice including Fc-receptor knockout mice. Opsonization will be investigated in murine macrophages in vitro with E. muris and specific polyclonal and monoclonal antibodies. Cellular immune mechanisms will be elucidated using gene knockout mice (MHC Class I, MHC Class II, delta T-cell receptor, IFN-gamma, perforin, iNOS, and TNF-alpha receptor) and TNF-alpha depleted mice, immunohistochemical and flow cytometric analyses of the cell subsets and their cytokine profiles, adoptive transfer of T-lymphocyte subsets including T-cell clones in the outstanding new mouse model of HME. Website: http://crisp.cit.nih.gov/crisp/Crisp_Query.Generate_Screen •

Project Title: EMERGING INFECTIOUS DISEASES AND URBANIZATION Principal Investigator & Institution: Ko, Albert I. Medicine; Weill Medical College of Cornell Univ New York, NY 10021 Timing: Fiscal Year 2003; Project Start 20-SEP-1997; Project End 31-MAR-2008 Summary: (provided by applicant): The Division of International Medicine and Infectious Disease, Weill Medical College of Cornell University (Cornell) has had a joint training and research program on endemic tropical diseases with Brazilian institutions in the city of Salvador since the 1964. More recently, the investigations of Cornell and its Brazilian collaborators have brought to attention infectious diseases, such as epidemic leptospirosis, which have emerged in the urban setting due to rapid urbanization and increasing social inequality. Through the Fogarty-sponsored International Training in Emerging Infectious Diseases (ITREID) Program, we have been established at the Oswaldo Cruz Foundation (Fiocruz), Brazilian Ministry of Health in Salvador: 1) a multidisciplinary team of epidemiologists, clinicians, microbiologists and basic researchers, 2) on-going population-based surveillance for leptospirosis and bacterial meningitis; 2) a diagnostic laboratory that is now the national reference center for leptospirosis surveillance; 3) a molecular strain typing center, and 4) field sites to perform community-based longitudinal studies designed to identify determinants of transmission for leptospirosis and the etiologic pathogens for bacterial meningitis. Moreover, ITREID projects have convinced the Brazilian government to prioritize emerging infectious diseases such as leptospirosis, and in turn have led to national projects to sequence the Leptospira genome and develop a vaccine against leptospirosis. The infrastructure created at Fiocruz since 1996 provides a vehicle to pursue multidisciplinary training approaches for emerging infectious diseases. In this proposal, we will use leptospirosis and bacterial meningitis as disease models to address the

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following specific objectives: 1) Expand training opportunities that will provide Brazilian trainees at Fiocruz the capacity to develop treatment, control and prevention strategies for emerging infectious diseases; and 2) Work jointly with the Brazilian Ministry of Health to disseminate expertise already established at Fiocruz in laboratorybased surveillance, outbreak investigations and molecular epidemiology to other regions of Brazil. The proposal emphasizes the use of in-country expertise and resources to provide training. For the first aim, we propose long-term training to 6 predoctoral and 2 postdoctoral fellows each year to address specific needs in expertise within the areas of clinical and field epidemiology, molecular epidemiology, pathogenesis and biotechnology application to develop public health interventions. In-country training provided by outstanding Brazilian mentors will be augmented with short training experiences in the institutions of long-standing US collaborators. For the second aim, Fiocruz will work with the National Center of Epidemiology, Brazilian Ministry of Health in providing short-term training opportunities and a yearly course, the National Course in Molecular Epidemiology in Emerging Infectious Diseases, which are designed to enhance the capacity of local public health epidemiologists and reference laboratory staff to perform laboratory-based surveillance and apply molecular strain typing tools to epidemiological investigations. Website: http://crisp.cit.nih.gov/crisp/Crisp_Query.Generate_Screen •

Project Title: ENTEROVIRUS 71 LEADER: TARGET FOR PEPTIDE INHIBITORS Principal Investigator & Institution: Thompson, Sunnie R. Microbiology and Immunology; Stanford University Stanford, CA 94305 Timing: Fiscal Year 2001; Project Start 01-MAR-2001 Summary: Every year hundreds of thousands of children are infected with enterovirus 71 (EV71). Infection with EV71 can lead to serious complications such as polio-like paralysis, encephalitis, meningitis or even death. Currently, there is no treatment or vaccine against EV71. EV71 belongs to the picornavirus family, whose members contain positive-stranded RNA genomes that are translated by an unusual mechanism of internal ribosome entry. The differences between the model of translational initiation of most cellular and viral mRNAs will be exploited to identify small peptides that can selectively inhibit the translation of the EV71 mRNA. Specifically, retroviruses expressing conformationally constrained peptide libraries will be used to infect cells expressing the EV71 5' non-coding region linked to an enhanced green fluorescent protein (EGFP) reporter gene and a cellular c-myc 5' non- coding region linked to an enhanced yellow fluorescence protein (EYFP). Infected cells which fail to express EGFP but still express EYFP will be isolated by cell sorting, and the gene encoding the putative inhibitor will be isolated. Studying the targets of the inhibitory peptides and characterizing the molecular interactions that they are disrupting will reveal more about how viruses function to recruit host cell molecules for their translation. Selection and characterization of intracellularly stable peptides that can inhibit the amplification of a viral RNA genome should level to novel ways in the search for antiviral therapeutics. Website: http://crisp.cit.nih.gov/crisp/Crisp_Query.Generate_Screen

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Project Title: ENTEROVIRUS RNA TRANSLATION AND REPLICATION Principal Investigator & Institution: Barton, David J. Assistant Professor; Microbiology; University of Colorado Hlth Sciences Ctr P.O. Box 6508, Grants and Contracts Aurora, CO 800450508 Timing: Fiscal Year 2003; Project Start 15-SEP-2003; Project End 31-AUG-2004

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Summary: (provided by applicant): Enteroviruses cause a diverse spectrum of human diseases including conjunctivitis, myocarditits, aseptic meningitis, acute flaccid paralysis, and fatal systemic infections in neonates. Poliovirus, the prototypic enterovirus, is well characterized at the molecular level and still serves as the most appropriate virus for studies of RNA translation and replication. In this proposal, poliovirus mRNA translation and RNA replication will be studied in cell-free reactions capable of supporting the sequential translation and replication of poliovirus RNA. These reactions are advantageous because they support authentic translation and replication of poliovirus RNA while providing numerous technical advantages including the ability to synchronize viral mRNA translation and viral RNA replication. The interaction of cis-active RNA structures at the termini of poliovirus RNA will be examined. Temporally dynamic ribonucleoproteins form on poliovirus cis-active RNA structures to mediate and regulate the sequential steps of replication. Experiments will be performed to: 1) determine how the 5' cloverleaf RNA structure of poliovirus potentiates viral mRNA translation, 2) determine how translating ribosomes regulate, in part, the switch from viral mRNA translation to RNA replication, 3) determine how apparently distal cis-active RNA structures interact to regulate sequential steps of viral RNA translation and replication, and 4) determine the mechanisms behind the asymmetric replication of poliovirus RNA. These experiments will help elucidate the fundamental sequence of molecular interactions required for enterovirus RNA translation and replication. This information will provide for a better understanding of the mechanisms by which enteroviruses replicate. In particular, these studies will contribute substantial new information to support the popular new paradigm of 5'-3' RNA interactions in messenger ribonucleoprotein complexes and RNA replication complexes. The experiments directly test a hypothesis concerning the mechanism by which RNA replication machinery avoids ribosome-replicase collisions. Finally, the experiments test a new model that clearly explains the mechanisms controlling asymmetric RNA replication. Website: http://crisp.cit.nih.gov/crisp/Crisp_Query.Generate_Screen •

Project Title: ESCHERICHIA COLI INVASION OF BRAIN ENDOTHELIAL CELLS Principal Investigator & Institution: Huang, Sheng-He; Children's Hospital Los Angeles 4650 Sunset Blvd Los Angeles, CA 90027 Timing: Fiscal Year 2001; Project Start 01-JUL-1997; Project End 30-JUN-2002 Summary: (Adapted from the applicant's abstract): Bacterial meningitis continues to be associated with high morbidity and mortality despite advances in chemotherapy and supportive care. E. coli is the most common gram-negative organism that causes neonatal meningitis. Most cases develop as a result of hematogenous spread, but it is not clear how circulating E. coli traverse the brain microvascular endothelium, which constitutes the blood brain barrier. The PI has demonstrated that the invasion of brain microvascular endothelial cells (BMEC) by E. coli K1 is mediated by multiple factors. Two separate genetic loci, identified by the PI as pathogenicity islands (paiA and paiB) apparently contribute to E. coli meningitis. PaiA contains the 20 kb E. coli locus identified as the major invasion gene cluster in K1 E. coli strain RS218. A TnphoA insertion mutant of E. coli RS218 was unable to invade BMEC in tissue culture and to cause hematogenous meningitis in a newborn rat model. Encoded on paiA is ibe10, which encodes an 8.2 kDa protein displaying the characteristics of an integral membrane protein with four transmembrane domains. A recombinant Ibe10 protein was able to block invasion of BMEC by E. coli K1. Ibe10 was detected in 30% of clinical isolates of K1 E. coli causing meningitis. Six clones for Ibe10 have been isolated from a lambda Gem-

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12 library. A recombinant plasmid carrying a 15 kb E. coli fragment was able to complement the non-invasive TnphoA mutant 10A-23. And, in an appendix, the PI reports that he has succeeded in conferring invasiveness on a strain of E. coli K12 using an 18 kb fragment of paiA. The long term goal of this work is to develop novel strategies for the prevention and treatment of E. coli meningitis (such as vaccines) based on a full understanding of the molecular mechanisms responsible for E. coli invasion of the blood-brain barrier. The PI hypothesizes that the phenotype of E. coli invasion of the blood-brain barrier is encoded by the pathogenic E. coli K1-specific genetic determinants, the pai's. The proposal comprises two aims: Aim 1: To further characterize the role of the invasion gene cluster paiA and/or paiB in the pathogenesis of E. coli meningitis by using in vitro (BMEC invasion) and in vivo (infant rat) models and molecular and genetic approaches. Aim 2: To determine the function and functional domains of invasion protein ibe10 by identification of the BMEC receptor for ibe10, in vitro mutagenesis and epitope mapping. Website: http://crisp.cit.nih.gov/crisp/Crisp_Query.Generate_Screen •

Project Title: FORMATION OF CORTICAL PLAQUES BY NEISSERIA Principal Investigator & Institution: So, Magdalene Y. Chair; Molecular Microbiol and Immun; Oregon Health & Science University Portland, OR 972393098 Timing: Fiscal Year 2001; Project Start 01-AUG-2001; Project End 31-MAY-2006 Summary: (provided by the applicant): The Neisseria type IV pilus modulates bacterial infectivity by mediating adhesion and inducing cell signaling pathways. Upon binding its receptor, CD46, the pilus causes a transient increase in cytosolic free Ca2+ levels, triggering endosome and lysosome exocytosis. Piliated bacteria next trigger elongation of microvilli and formation of cortical plaques at the plasma membrane beneath the site of contact. Cortical plaques contain clusters of Opa receptors, transmembrane signaling proteins, actin microfilaments and ezrin, a protein that tethers the membrane to the actin cytoskeleton. These plaques serve multiple signaling functions that promote bacterial infection. Unlike the Ca2+ response, which can be induced with purified pill, cortical plaque formation requires live diplococci and PilT, a protein that functions in pilus assembly and DNA transformation. PilT also controls pilus retraction, a process that drives twitching motility and the ability of diplococci to aggregate into microcolonies. Retraction generates substantial force on the substrate to which the pilus is attached. External forces placed on the membranes of eucaryotic cells result in the induction of kinase cascades, cytoskeleton reorganization and alterations in translation. The tension generated on the plasma membrane by retraction of pili during infection may therefore act as a signal to promote the formation of cortical plaques and subsequent bacterial invasion. Based on these and other observations, we propose a model for early events in pilus-induced cortical plaque formation. In this proposal, we describe experiments to test key predictions of this model. Website: http://crisp.cit.nih.gov/crisp/Crisp_Query.Generate_Screen

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Project Title: FREQUENCY DOMAIN CEREBRAL OXIMETER FOR PEDIATRICS Principal Investigator & Institution: Kurth, Dean C. Attending Anesthesiologist; Near Infrared Monitoring, Inc. (Nim) 3401 Market St, Ste 140F Philadelphia, PA 19104 Timing: Fiscal Year 2001; Project Start 01-APR-2000; Project End 29-SEP-2002 Summary: Brain damage from hypoxia-ischemia represents a major health problem in pediatrics. At present, no method exists in clinical care to diagnose cerebral hypoxiaischemia in real-time at the bedside, thereby limiting prevention and treatment of the

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brain damage. Near infrared spectroscopy (cerebral oximetry) is an emerging optical technology with the potential to fulfill this role. In previous work, we built a prototype frequency domain near infrared cerebral oximeter and found it measures cerebral O2 saturation accurately. However, before clinicians will use cerebral O2 saturation to diagnose cerebral hypoxia-ischemia, the measure needs to be related to other known measures of hypoxia-ischemia, and the instrument needs to be engineered to the clinical environment. This fast-track proposal will develop a frequency domain near infrared cerebral oximeter to diagnose cerebral hypoxia-ischemia in real-time at the bedside for pediatrics (