EPSIS 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 2004 by ICON Group International, Inc. Copyright 2004 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., 1960Sepsis: 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: 0-597-84623-5 1. Sepsis-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.
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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 sepsis. 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 SEPSIS ........................................................................................................ 3 Overview........................................................................................................................................ 3 The Combined Health Information Database................................................................................. 3 Federally Funded Research on Sepsis............................................................................................. 5 E-Journals: PubMed Central ....................................................................................................... 62 The National Library of Medicine: PubMed ................................................................................ 71 CHAPTER 2. NUTRITION AND SEPSIS ............................................................................................ 115 Overview.................................................................................................................................... 115 Finding Nutrition Studies on Sepsis ......................................................................................... 115 Federal Resources on Nutrition ................................................................................................. 118 Additional Web Resources ......................................................................................................... 119 CHAPTER 3. DISSERTATIONS ON SEPSIS ........................................................................................ 121 Overview.................................................................................................................................... 121 Dissertations on Sepsis .............................................................................................................. 121 Keeping Current ........................................................................................................................ 121 CHAPTER 4. CLINICAL TRIALS AND SEPSIS ................................................................................... 123 Overview.................................................................................................................................... 123 Recent Trials on Sepsis .............................................................................................................. 123 Keeping Current on Clinical Trials ........................................................................................... 131 CHAPTER 5. PATENTS ON SEPSIS ................................................................................................... 133 Overview.................................................................................................................................... 133 Patents on Sepsis........................................................................................................................ 133 Patent Applications on Sepsis.................................................................................................... 154 Keeping Current ........................................................................................................................ 188 CHAPTER 6. BOOKS ON SEPSIS ...................................................................................................... 189 Overview.................................................................................................................................... 189 Book Summaries: Federal Agencies............................................................................................ 189 Book Summaries: Online Booksellers......................................................................................... 191 Chapters on Sepsis ..................................................................................................................... 195 CHAPTER 7. MULTIMEDIA ON SEPSIS ............................................................................................ 205 Overview.................................................................................................................................... 205 Audio Recordings....................................................................................................................... 205 CHAPTER 8. PERIODICALS AND NEWS ON SEPSIS ......................................................................... 207 Overview.................................................................................................................................... 207 News Services and Press Releases.............................................................................................. 207 Newsletter Articles .................................................................................................................... 211 Academic Periodicals covering Sepsis........................................................................................ 212 CHAPTER 9. RESEARCHING MEDICATIONS .................................................................................. 213 Overview.................................................................................................................................... 213 U.S. Pharmacopeia..................................................................................................................... 213 Commercial Databases ............................................................................................................... 214 Researching Orphan Drugs ....................................................................................................... 215 APPENDIX A. PHYSICIAN RESOURCES .......................................................................................... 219 Overview.................................................................................................................................... 219 NIH Guidelines.......................................................................................................................... 219 NIH Databases........................................................................................................................... 221 Other Commercial Databases..................................................................................................... 223 APPENDIX B. PATIENT RESOURCES ............................................................................................... 225 Overview.................................................................................................................................... 225 Patient Guideline Sources.......................................................................................................... 225
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Finding Associations.................................................................................................................. 229 APPENDIX C. FINDING MEDICAL LIBRARIES ................................................................................ 231 Overview.................................................................................................................................... 231 Preparation................................................................................................................................. 231 Finding a Local Medical Library................................................................................................ 231 Medical Libraries in the U.S. and Canada ................................................................................. 231 ONLINE GLOSSARIES................................................................................................................ 237 Online Dictionary Directories ................................................................................................... 242 SEPSIS DICTIONARY.................................................................................................................. 243 INDEX .............................................................................................................................................. 343
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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 sepsis is indexed in search engines, such as www.google.com or others, a nonsystematic 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 sepsis, 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 sepsis, 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 sepsis. 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 sepsis, 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 sepsis. 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 SEPSIS Overview In this chapter, we will show you how to locate peer-reviewed references and studies on sepsis.
The Combined Health Information Database The Combined Health Information Database summarizes studies across numerous federal agencies. To limit your investigation to research studies and sepsis, 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 “sepsis” (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: •
Prevention and Treatment of Acute Renal Failure in Sepsis Source: JASN. Journal of the American Society of Nephrology. 14(3): 792-805 March 2003. Contact: Available from Lippincott Williams and Wilkins. 12107 Insurance Way, Hagerstown, MD 21740. (800) 638-6423. Website: www.jasn.org/. Summary: Acute renal failure (ARF) is a common complication of sepsis and has a poor prognosis. Mortality (death) was reported higher in patients with septic ARF (74.5 percent) than in those whose renal (kidney) failure did not result from sepsis (45.2 percent). This article discusses the use of drug therapy to interfere with each of the dysfunctional pathways to improve the course of septic ARF. These include inhibition of inflammatory mediators, improvement of renal hemodynamics (blood flow) by
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amplifying vasodilator mechanisms and blocking vasoconstrictor mechanisms, interruption of leukocyte infiltration, inhibition of the coagulation cascade, and administration of growth factors to accelerate renal recovery. The author also highlights the available supportive measures, including dialysis, that can be used for septic patients with ARF. The author notes that, unfortunately, treatment of ARF in sepsis is still only supportive; there have been no conclusive drug therapies available to treat the condition. 1 figure. 1 table. 136 references. •
Urosepsis: When to Suspect, How to Confirm, How to Manage Source: Journal of Critical Illness. 12(10): 616-618, 623-625. October 1997. Contact: Available from Cliggott Publishing Company. 55 Holly Hill Lane, Greenwich, CT 06831-0010. (203) 661-0600. Summary: This article reminds physicians of the recommended care for patients with urosepsis (bacteremia that originates in the urinary tract). Urosepsis can be acquired in the hospital (nosocomial) or in the community. The mere presence of bacteriuria is not sufficient to diagnose urosepsis in patients who have symptoms of sepsis. Urosepsis develops only in specific clinical situations: when host defenses are impaired (as they are in patients who have diabetes mellitus or systemic lupus erythematosus) or when they are circumvented or bypassed by mechanical factors (such as obstruction or preexisting renal disease). To confirm the diagnosis of urosepsis, the same pathogen found in the urine must be found in the blood; in addition, the pathogen must be linked to a site of infection in the genitourinary tract or to urologic instrumentation or catheterization. The author stresses that knowing the source of urosepsis helps physicians to determine the most likely causative organisms and therefore the most appropriate antibiotic therapies. Nosocomial urosepsis usually results from either urologic manipulation or an indwelling Foley catheter in nonleukopenic, immunocompromised patients. Community-acquired urosepsis may complicate any type of urinary tract infection, but only in patients with preexisting renal disease, obstruction, or impaired host defenses. If urosepsis is related to obstruction, antibiotics can temporarily control the infection process, but they cannot eliminate it. Whenever possible, urologic or surgical correction of the obstruction should be performed. (AAM).
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Life-Threatening Retroperitoneal Sepsis After Hemorrhoid Injection Sclerotherapy: Report of a Case Source: Diseases of the Colon and Rectum. 42(3): 421-423. March 1999. Contact: Available from Williams and Wilkins. 352 West Camden Street, Baltimore, MD 21201-2436. Summary: This article reports a case of life threatening retroperitoneal sepsis after injection sclerotherapy for first degree hemorrhoids. A 50 year old man with symptomatic first degree hemorrhoids was seen in the outpatient department. An experienced surgical registrar injected three internal hemorrhoids with 3 to 5 mL of 5 percent oily phenol. Four days later, the patient was admitted as an emergency, complaining of tight central chest pain. For 6 hours he had experienced chills and pelvic pain radiating to his lower abdomen and the backs of his thighs. Anorectal instrumentation was not attempted, because the patient was reporting severe pelvic pain. Investigation revealed leukocytosis, raised creatinine kinase, and electrocardiographic changes suggestive of anteroseptal myocardial infarction. Streptokinase, cefotaxime, and metronidazole were administered. Later that day, his
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pelvic pain worsened. He remained pyrexial (having a fever), developed tachycardia (rapid heartbeat), and went into urinary retention. Computed tomography revealed extensive retroperitoneal fluid but no localized abscess. Blood culture isolated gram negative bacilli, but exploratory laparotomy found no colonic lesion. The fecal stream was diverted with an end sigmoid colostomy and the rectal stump was oversewn. Hyperbaric oxygen, antibiotics, and intensive inotropic and ventilatory support were continued in the postoperative period. The patient eventually made a good recovery. The authors note that life threatening sepsis after injection sclerotherapy for hemorrhoids has been reported only once previously. One table summarizes the cases of life threatening complications after rubber band ligation of hemorrhoids. 1 table. 9 references.
Federally Funded Research on Sepsis The U.S. Government supports a variety of research studies relating to sepsis. 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. 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 sepsis. 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 sepsis. The following is typical of the type of information found when searching the CRISP database for sepsis: •
Project Title: ACUTE LUNG INJURY--ALCOHOLISM AND GLUTATHIONE DEPLETION Principal Investigator & Institution: Guidot, David M.; Associate Professor of Medicine; Medicine; Emory University 1784 North Decatur Road Atlanta, Ga 30322 Timing: Fiscal Year 2002; Project Start 01-APR-1999; Project End 31-MAR-2003 Summary: The Acute Respiratory Distress Syndrome (ARDS) is a common and severe form of lung injury with a mortality of approximately 50 percent. A prospective study of 351 critically ill patients recently identified that a history of chronic alcohol abuse increased the incidence and severity of ARDS regardless of the at-risk diagnosis. This observation distinguishes chronic alcohol abuse as the first reported co-morbid variable that significantly increases a patient's risk of developing ARDS and raises questions about the pathophysiology and specific treatment of acute lung injury. This project will test the hypothesis that chronic alcohol abuse decreases alveolar type II cell levels of glutathione, an important antioxidant, thereby impairing surfactant secretion and function and rendering the lung susceptible to injury. In ARDS the alveolar type II cells are severely damaged, and their ability to secrete glutathione and surfactant into the
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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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alveolar lining fluid are critical to patient survival. Because sepsis is the most common risk factor for ARDS, this project will focus on the sepsis syndrome in both patients and in an animal model. Preliminary studies presented in this proposal show that chronic alcohol ingestion in rats decreases type II cell glutathione levels and, in parallel, decreases type II cell surfactant secretion both in vitro and in vivo, and predisposes to endotoxin-mediated acute lung injury. In addition, we determined that otherwise healthy alcoholics have markedly decreased levels of glutathione in their lung lavage fluid compared to control subjects. The fundamental mechanisms by which chronic alcohol use affects alveolar epithelial function will be examined in a rat model of sepsis in vivo and in isolated type II cells in vitro. Parallel clinical studies in both healthy subjects and in critically ill patients with sepsis will examine the effects of chronic alcohol abuse on lung glutathione homeostasis and surfactant production both in isolated type II cells and in lung lavage fluid. We will thereby test the clinical relevance of the fundamental mechanisms elucidated in the animal model. Importantly, our preliminary studies indicate that glutathione replacement can decrease ethanolmediated lung injury in our animal model, and this project will ultimately focus on developing a glutathione replacement regimen that reduces the harmful effects of chronic alcohol ingestion on the lungs of patients. Website: http://crisp.cit.nih.gov/crisp/Crisp_Query.Generate_Screen •
Project Title: ACUTE LUNG INJURY-MECHANISMS AND THERAPY Principal Investigator & Institution: Crapo, James D.; Executive Vice President for Academic Af; National Jewish Medical & Res Ctr and Research Center Denver, Co 80206 Timing: Fiscal Year 2002; Project Start 01-MAY-1984; Project End 31-MAR-2005 Summary: PROPOSED PROGRAM (Adapted from Applicant's Abstract) The long-term objectives of this Program Project application are to evaluate basic mechanisms and develop new treatments for acute lung injury. High concentrations of oxygen and septic lung injury are the primary models that will be evaluated. The proposed program consists of four projects and three core units. Project 1 will evaluate the efficacy of small molecular weight catalytic antioxidants in the treatment of both hyperoxic and LPS + sepsis-initiated lung injury. This project will also develop new antioxidant mimetics and explore their relationships with the antioxidant properties of heme oxygenase (HO). Project 2 will test the hypothesis that activation of extrinsic coagulation and disordered fibrin turnover are central elements in hyperoxic and septic lung injury. The efficacy of specific blockade of the initiating steps of extrinsic coagulation in reducing inflammation and acute lung injury will be tested using two new anticoagulant drugs that block tissue factor (TF) function and do not cause bleeding. Project 3 will evaluate the regulation and function of the extracellular superoxide dismutase (EC-SOD) in acute lung injury and determine the impact of cleavage of the C-terminal "heparin binding" domain of this enzyme in determining its distribution and function. Project 4 will evaluate control of metabolic pathways and upregulation of lung cell glycolysis in modulating responses to acute injury. This project will test the hypothesis that adaptation to oxidant stress in the lung requires elevated expression of hexokinase (HK), a rate limiting step in glycolysis in the lung. The overall rationale for the Program Project is to use an interdisciplinary approach to define the cellular pathways and cellular adaptive responses involved in acute lung injury and to test new strategies for pharmacologic therapy that can be extended to the treatment of humans with ARDS and sepsis. Website: http://crisp.cit.nih.gov/crisp/Crisp_Query.Generate_Screen
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Project Title: AMELIORATION OF SEPSIS BY MACROPHAGE ACTIVATION Principal Investigator & Institution: Williams, David L.; Professor; Surgery; East Tennessee State University Box 70565 Johnson City, Tn 37601 Timing: Fiscal Year 2002; Project Start 01-APR-1996; Project End 31-JAN-2005 Summary: During the previous grant period our research focused on understanding the early intracellular signaling events which are involved in the initiation and propagation of sepsis syndrome. We found that suppressing pro-inflammatory intracellular signaling events during the early phase of sepsis strongly correlated with improved outcome. Specifically, we investigated the protective efficacy of glucan ligands in polymicrobial sepsis and determined that receptor recognition of glucan modulates intracellular signaling pathways such that the inflammatory response to sepsis is "blunted". We established that glucan pre- or post-treatment would blunt sepsis induced tissue NFkappaB and NF-IL6 activation as well as decrease pro-inflammatory cytokine gene transcription. Blunting early increases in transcription factor activity and cytokine gene expression strongly correlated with decreased morbidity and mortality. We determined that glucan treatment blunted LPS induced NFkappaB activity through decreased MEKK1, NIK and IKKalpha/beta kinase activity as well as decreased IkappaBalpha phosphorylation and degradation. We confirmed the existence of multiple glucan binding sites on macrophages, defined the molecular structure of a glucan and confirmed that a heptaose (7 glucose subunit) polymer was the minimum binding unit. Preliminary data suggest that Toll receptor (TLR) 2, and perhaps CR3, confers responsiveness to glucan. The hypothesis for this continuation proposal is that lucan ligands ameliorate septic sequelae in polymicrobial sepsis by modulating inflammatory responses via interactions with Toll-like receptors (TLR) and/or CR3 (CD11b/CD18) binding sites. There are four specific aims. 1. We will characterize the receptor mediated interaction of glucan ligands with TLR 1, 2 and 4, the type 3 complement receptor (CR3) and CD 14 using a surface plasmon resonance approach. 2. We will establish the role of TLR2 and TLR4 in the anti-sepsis effect of glucan by studying CLP sepsis in TLR2 and TLR4 knockout mice. 3. We will investigate the role of CR3 (CD11b/CD18) in the antisepsis effect of glucan by studying CLP sepsis in CR3 knockout mice. 4. During the last grant period we made great strides in understanding the basic chemistry of (1-3)-beta-Dglucans. Using this knowledge, we will synthesize chemically pure, highly uniform, water soluble (1-3)-beta-D-glucan ligands which have specific structural characteristics. We will prepare a library of small molecular weight (1-3)-beta-D-glucans which will be used to prepare larger polymers. The synthetic polymers will be evaluated in receptor binding studies, in vitro intracellular signaling studies and in vivo protection studies using the CLP model. The successful completion of these aims will advance our understanding of the cellular and molecular events associated with sepsis syndrome and may lead to the development of new therapeutics. Website: http://crisp.cit.nih.gov/crisp/Crisp_Query.Generate_Screen
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Project Title: APOPTOSIS IN SEPSIS: REGULATION BY TNF & DELTA-PKC Principal Investigator & Institution: Kilpatrick, Laurie E.; Associate Member; Children's Hospital of Philadelphia 34Th St and Civic Ctr Blvd Philadelphia, Pa 191044399 Timing: Fiscal Year 2002; Project Start 10-JAN-2002; Project End 31-DEC-2006 Summary: Spontaneous apoptosis of neutrophils is attenuated during sepsis and inflammatory cytokines such as TNFalpha have been implicated. The pathophysiological mechanisms involved in TNFalpha-mediated attenuation of apoptosis are poorly understood but proposed to be mediated by the p60 TNF receptor
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(p60TNFR). In adherent neutrophils to engage beta2 integrins, TNFalpha triggers phosphorylation of the p60TNFR, activation of cell survival signaling pathways and inhibition of spontaneous apoptosis. Our studies implicate phosphatidylinositol 3kinase (PI 3-kinase) in TNFalpha triggered NFkappaB activation and that TNFalpha activation of PI 3-kinase requires beta2 integrins. The protein kinase C isotype delta (delta-PKC) phosphorylates p60TNFR on serine residue(s) in TNFalpha activated neutrophils. Rottlerin, a delta-PKC inhibitor, suppressed the inhibitory effect of TNFalpha on neutrophil apoptosis and activation of the transcription factor NFkappaB suggesting that delta-PKC may regulate TNFalpha anti-apoptotic signaling. Our model proposes a selective role for delta-PKC in regulating anti-apoptotic signaling triggered by TNFalpha binding to the p60TNFR. The goal of this study is to establish a role for delta-PKC in TNFalpha mediated anti-apoptotic signaling through the p60TNFR using antisense technology to selectively deplete delta-PKC from HL60 cells differentiated to a neutrophilic phenotype. We will: 1: Determine whether delta-PKC regulates TNFalpha mediated anti- apoptotic signaling. Determine the effect of delta-PKC deletion on TNFalpha-mediated suppression of spontaneous apoptosis and activation of the antiapoptotic NFkappaB and the MAP kinases ERK1/2. 2: Assess the role of PI 3-kinase in TNFalpha mediated suppression of spontaneous apoptosis, and determine if deltaPKC is required for TNFalpha mediated activation of PI 3-kinase. 3: Identify the deltaPKC phosphorylation site on the p60TNFR and establish if phosphorylation is in the a) death domain, b) juxtamembrane region, or c) a novel domain. 4: Assess the role of delta-PKC in regulating the assembly of TNFalpha anti-apoptotic signaling complexes and their association with the p60TNFR. Website: http://crisp.cit.nih.gov/crisp/Crisp_Query.Generate_Screen •
Project Title: BACTERIAL SURFACE PROTEINS: POTENTIAL TARGETS FOR SEPSIS Principal Investigator & Institution: Hellman, Judith; Massachusetts General Hospital 55 Fruit St Boston, Ma 02114 Timing: Fiscal Year 2002; Project Start 01-JUN-2000; Project End 31-MAY-2005 Summary: Bacterial cell wall components released into the bloodstream are believed to be important in eliciting the inflammatory response that causes Gram-negative Sepsis. The bacterial outer membrane contains lipopolysaccharide (LPS), outer membrane proteins (OMPs), and phospholipids. LPS has been studied extensively as an important mediator in sepsis; the importance of OMPs in sepsis has not been systematically evaluated. Our preliminary data indicate that complexes containing three conserved OMPs and LPS are released from bacteria into human serum and are also released into the circulation of septic rats. We have identified the three proteins and have found that at least one of the proteins has biological activity. The major goals of this project are to further characterize release of the three OMPs in sepsis, to assess their roles in the pathogenesis of sepsis, and to evaluate anti-OMP immunity as a means of treating sepsis. The first specific aim is to assess release of OMPs and OMP/LPS complexes in humans with Gram- negative sepsis and in two models of experimental Gram-negative sepsis in rats. The second specific aim is to evaluate individual OMPs, OMP/LPS complexes, and OMPs on the surface of intact bacteria for biological activity in vitro on isolated immune cells, an in vivo by studying lethality, induction of cytokines in blood and organs, and pulmonary neutrophil sequestration in LPS resistant and LPS responsive mice. The third specific aim is to evaluate protective effects of active immunity on toxicity of OMP containing preparations in mice and in the rat infection models of sepsis. The applicant's long-term career objective is to become an
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independent investigator studying basic mechanisms of Gram- negative infection and sepsis. The career development plan is designed to expand the candidate's research knowledge base through hands-on research and educational activities. These will include course work (laboratory and didactic), attendance at and participation in laboratory meetings and research seminars, and formal and informal interactions with the Advisory Committee members. The ultimate goal of the candidate is to apply the fundamental knowledge of mechanisms involved in sepsis to develop anti-sepsis strategies that target bacterial toxins. Website: http://crisp.cit.nih.gov/crisp/Crisp_Query.Generate_Screen •
Project Title: INFECTIONS
CAPSULAR
POLYSACCHARIDES
AND
ENTEROCOCCAL
Principal Investigator & Institution: Huebner, Johannes; Brigham and Women's Hospital 75 Francis Street Boston, Ma 02115 Timing: Fiscal Year 2002; Project Start 01-SEP-1998; Project End 31-JUL-2002 Summary: (Adapted from applicant's abstract): The long-term goal of this study is to understand the molecular, biochemical, and immunologic factors that contribute to the pathogenesis of Enterococcus faecalis and E.faecium infections, with special emphasis on multiply antibiotic-resistant enterococci. Further understand of the interaction of these bacteria with the human host defense system will give new insights into how to prevent disease due to enterococcal infections. The molecular and biochemical studies will concentrate on isolating and characterizing likely capsular polysaccharides of enterococci which will culminate in deterring the monosaccharide components and linkages in these antigens. Electron microscopy using antibodies specific to isolated polysaccharides will determine if the materials are extracellular capsules. The immunologic studies will focus on whether the isolated antigens are targets for protective immune responses. In this study between three and five serologically distinct antigens will be purified and characterized chemically and immunologically. A mouse peritonitis/sepsis model and a neonatal mouse model will be used to evaluate the effectiveness of antibodies specific for the capsular polysaccharides in prevention of enterococcal infections. Active immunization with purified capsule and passive protection using capsule-specific antibodies will be tested in this models. By the end of these studies we expect to have a clear understanding of the occurrence, biochemistry and immunology of capsular polysaccharides of enterococci, a definition of their role in pathogenesis of enterococcal infections, and the potential to develop immunotherapies to prevent infection in patient at-risk for this disease. Website: http://crisp.cit.nih.gov/crisp/Crisp_Query.Generate_Screen •
Project Title: CHARACTERIZATION OF IRON TRANSPORT IN GBS Principal Investigator & Institution: Clancy, Kathryn A.; Children's Hospital and Reg Medical Ctr Box 5371, 4800 Sand Point Way Ne, Ms 6D-1 Seattle, Wa 98105 Timing: Fiscal Year 2003; Project Start 01-APR-2003; Project End 31-MAR-2005 Summary: (provided by applicant): Group B streptococcus is the predominant etiological agent of neonatal bacteremia, sepsis and meningitis, and has been correlated with preterm membrane rupture and premature birth. While a limited number of GBS virulence determinants have been identified, the contribution of iron and iron acquisition to the pathogenesis of GBS infection is unknown. All pathogens require iron and a correlation between bacterial virulence and iron acquisition has been established. However, as the concentration of available iron in the human host (10 [-18]M) is well
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below that required by bacteria (10 [-8] M), successful pathogens express specific systems to acquire sufficient concentrations of intracellular iron. One such system involved in siderophore-mediated iron uptake, is the focus of this application. Siderophores are high-affinity iron chelators secreted from the bacterial cell to scavenge iron from host iron-binding proteins. Specialized uptake systems transport the siderophore-iron complex across the bacterial membrane. We have identified a putative siderophore-mediated iron transport system, the fhu operon, in GBS. The operon is comprised of four genes, fhuC, fhuD, fhuB, and fhuG, encoding a putative ATPhydrolysis protein, siderophore (ferrichrome)- iron receptor protein, and two permeases, respectively. In this application, the role of the fhu operon in iron acquisition by GBS will be examined. Basic information on the requirement of GBS for iron, the ability of GBS to utilize siderophores as an iron source, and whether the organism secretes siderophores to acquire iron will first be established. The biochemical characterization of an isogenic mutant strain deficient for fhu will examine the role of this operon in GBS siderophore-iron transport. In order to define the siderophore specificity of the fhu operon, the construction and biochemical characterization of isogenic mutant strains deficient for each gene is proposed. In addition, heterologous expression studies will provide further evidence for the role of the fhu-encoded proteins in siderophore-iron transport. These studies will provide a basic understanding of the molecular mechanism of iron acquisition in GBS and will allow us to design appropriate in vivo studies to determine the role of iron transport in GBS pathogenesis. The potential of identifying novel therapeutic targets to prevent or treat infections is also proposed. Website: http://crisp.cit.nih.gov/crisp/Crisp_Query.Generate_Screen •
Project Title: CHARACTERIZATION OF THE LPS RECEPTOR FOR ACUTE PHASE Principal Investigator & Institution: Goyert, Sanna M.; Associate Professor; North Shore University Hospital 300 Community Dr Manhasset, Ny 11030 Timing: Fiscal Year 2002; Project Start 01-JUN-2000; Project End 31-MAY-2004 Summary: Sepsis due to Gram-negative infection remains a major cause of mortality. One of the earliest events occurring in a systemic infection is the acute phase response which has, as one of its major hallmarks, alteration of the concentration of plasma proteins (acute phase proteins, APP). APP are a set of functionally diverse proteins produced in the liver and defined in general as those proteins which show changes in plasma concentration (positive or negative) of 25 percent or more following the stimulus. APP are thought to increase host defenses as well as to control inflammation. There is a large body of evidence showing that cytokines (TNFalpha, IL-1 and IL-6) can induce the expression of APP. Since lipopolysaccharide (LPS, endotoxin), a component of the outer membrane of Gram-negative bacteria which is thought to be the major bacterial component of Gram-negative bacteria responsible for inducing the cascade of events leading to lethality in sepsis, stimulates both the production of TNFalpha, IL-1 and IL-6 as well as the production of APP, it has been reasonable to assume that the LPS induction of APP results from a secondary effect of cytokines secreted by macrophages when LPS stimulates them through the CD14-LPS receptor. To study the role of CD14 in the response to LPS, we have recently produced mice which lack the CD14-LPS receptor. These CD14-deficient mice produce little or no cytokines in response to very high concentrations of LPS; surprisingly however, they have a normal APP response. These observations indicate that mice have a non-CD14 receptor for LPS through which expression of APP is induced. Furthermore, as shown in Preliminary Studies, hepatocytes from CD14-deficient mice respond directly to LPS, indicating that this
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11
receptor is on hepatocytes. Accordingly, we propose to (1) study the binding characteristics of LPS and Lipid A to hepatocytes; determine whether their binding is specific and saturable; determine their binding constants (2) isolate and biochemically characterize the hepatocyte non-CD14 LPS receptor involved in the induction of the acute phase proteins using molecular methods of protein purification and gene cloning followed by functional verification and (2) determine the molecular mechanism(s) by which the LPS-APP receptor functions by comparing the genes induced via this receptor in hepatocytes to those induced by LPS via the CD14 receptor on monocytes/macrophages. These studies will not only clarify our understanding of the mechanisms involved in the induction of acute phase proteins by LPS, but will also increase our understanding of the pleiotropic effects of LPS and its various roles in sepsis. Website: http://crisp.cit.nih.gov/crisp/Crisp_Query.Generate_Screen •
Project Title: CORE--FORWARD GENETICS Principal Investigator & Institution: Beutler, Bruce; Professor; Scripps Research Institute Tpc7 La Jolla, Ca 92037 Timing: Fiscal Year 2002; Project Start 30-SEP-2002; Project End 31-JUL-2007 Summary: (provided by applicant): While several of the genes that support the innate immune response have been identified through studies of natural mutations or gene knockouts, the great majority of these genes remain undiscovered. The Forward Genetics Core Laboratory will implement the goal of identifying most or perhaps all of the genes that subserve innate immunity. This goal will be achieved through the concerted production of novel mutations that impair innate immunity and through the mapping and isolation of those mutations that are created. Saturation mutagenesis will be accomplished in mice using N-ethyl-N-nitrosourea (ENU), a potent chemical agent that is known to produce chiefly A to G and A to T substitutions. Animals bearing mutations will be screened for phenotypic impairment of innate immune responses at the FI and F3 generations, to identify both dominant and recessive mutations of genes that are critical for innate immune responses. One screen to be employed relies upon a test of the ability to resolve infection of a type that is known to be contained entirely through innate immune mechanisms. In this acute model, a photometrically based assay of sepsis that depends upon extracorporeal detection of bacterial luminescence will be employed. Other screens measure the ability of host phagocytes to engulf and to kill microbes; the ability to respond to a battery of microbial inducers (PAMPs), and the ability to develop a state of tolerance to these inducers. Hence, a broad array of genes is tested, encompassing much of the functionality of the innate immune system. The Forward Genetics Core will interact particularly strongly with the Bioinformatics Core (which will pinpoint additional genes that appear to be involved in the innate immune response), the Molecular Biology Core, which will actively study mutations that are induced. Perhaps foremost, the Forward Genetics Core Laboratory will yield a rich harvest of mutants that will permit extensive genetic dissection of innate immune responses, and their failure in septic shock. These mutants will immediately be made available to the scientific community at large and simultaneously refined for positional cloning. Website: http://crisp.cit.nih.gov/crisp/Crisp_Query.Generate_Screen
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Project Title: CORE--GENOMICS LABORATRORY Principal Investigator & Institution: Aderem, Alan A.; Professor & Head; Institute for Systems Biology 1441 N 34Th St Seattle, Wa 981038904
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Sepsis
Timing: Fiscal Year 2002; Project Start 30-SEP-2002; Project End 31-JUL-2007 Summary: (provided by applicant): The analysis of genetic information will be a central requirement for all of the projects described in this grant proposal. The Genomics Core will provide high-throughput facilities for DNA sequencing, mRNA expression analysis, and single-nucleotide polymorphism (SNP) detection at the Institute for Systems Biology (ISB). The sequencing core is a high-capacity, fully automated facility developed to manage sequence analysis from eDNA libraries, BAC libraries, or PCR products in a cost-effective manner. The in-house array production and analysis facility at the ISB will be used to quantitate mRNA expression of genes involved in sepsis. This facility is currently producing high-density arrays (up to 40,000 genes per array) as well as customized arrays with select sets of genes. The facility is automated to analyze multiple arrays from clinical samples, which will be necessary to correlate in vitro data with clinical data. SNP technology has been established to both discover new SNPs as well as provide high-throughput SNP detection from multiple patient samples. SNPs are discovered by a heteroduplex mismatch detection method that has been adapted for use with arrays in a high-throughput manner. Known SNPs are detected by hybridization of an allele-specific oligonucleotide probe to an array of PCR products amplified from patient's genomic DNA. Website: http://crisp.cit.nih.gov/crisp/Crisp_Query.Generate_Screen •
Project Title: CORE--PROTEOMICS Principal Investigator & Institution: Aebersold, Ruedi H.; Professor and Co-Founder; Institute for Systems Biology 1441 N 34Th St Seattle, Wa 981038904 Timing: Fiscal Year 2002; Project Start 30-SEP-2002; Project End 31-JUL-2007 Summary: (provided by applicant): This core will facilitate studies in most of the collaborative projects and cores by providing expert and high throughput protein analysis for identification and quantification of proteins in complex samples. The core utilizes biochemical protein separation techniques and mass spectrometry as the primary technologies to achieve this goal, and members of this core facility are world leaders in this field. It is the aim of this core to establish a discovery science infrastructure for qualitative and quantitative proteomics at the Institute of Systems Biology. Through interactive collaborations with Glue Grant investigators this core will impact biomedical research in the regulation of the immune response during injury and sepsis. This core facility will permit participants in this grant proposal to 1) rapidly identify unknown proteins, 2) characterize specific post-translational modifications, 3) identify interacting proteins 4) localize protein:protein interaction sites, and 5) quantify the relative abundance of up to thousands of different proteins in complex specimens. Website: http://crisp.cit.nih.gov/crisp/Crisp_Query.Generate_Screen
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Project Title: DESIGN OF ATTENUATED TULAREMIA VACCINE Principal Investigator & Institution: Cross, Andrew M.; University of Maryland Balt Prof School Baltimore, Md 21201 Timing: Fiscal Year 2003; Project Start 01-AUG-2003; Project End 31-JUL-2008 Summary: Its ease of transmission, history of having been weaponized and propensity to cause severe and fatal disease following inhalation, make Francisella tularensis (Ft) a Category A bioterrorism agent of concern. The only vaccine available for more than 40 years is efficacious, but its mode of attenuation is unknown and the FDA has not approved its general use. Development of new vaccines is limited by the paucity of information about the virulence determinants of Ft. This project will provide (1)
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measures to extend the disease-free interval until definitive therapy/vaccines are implemented; and (2) vaccines that induce humoral and cellular immunity to Ft. Project 1A will characterize mechanisms by which the unusual Ft LPS induces mediators responsible for the systemic inflammatory responses of tularemia, and determine if reagents already under clinical testing for sepsis are useful in the treatment of disseminated tularemia in a murine model. Upon stimulation, gamma-delta T cells rapidly produce inflammatory cytokines critical to both the initial innate immune response and organization of the adaptive responses. Activation of gamma-delta T cells is associated with convalescence from tularemia. Aminobisphosphonates drugs, widely used for bone disorders, stimulate gamma-delta T cells and might serve as initial therapy for individuals exposed to Ft (Project 1B). Project 2 will characterize the Ft capsule and develop a conjugate vaccine, using as carriers either the protective antigen of B. anthracis or proteins derived from plague or Ft. Adjuvants that also rapidly boost innate immunity (e.g. CpG) may accelerate a humoral response and provide early protection. Like the Vi vaccine for the intracellular pathogen, Salmonella Typhi, the Ft capsular conjugate vaccine is intended to prevent Ft from reaching its required intracellular niche. Durable immunity to Ft requires a cellular immune response. Based on our previous success in developing live attenuated strains of Salmonella, we will design an attenuated, easily administered Ft vaccine (Project 3A). Signature-tagged mutagenesis will define additional targets for attenuation and new virulence factors for further study (Project 3C). Activated T cells are sequestered in peripheral tissues. We will compare which immunization regimen optimally delivers primed effector/memory T cells to lung and liver, sites of Ft replication. These studies will provide public health officials short term and definitive treatment options in the event of a bioterror attack with Ft. Website: http://crisp.cit.nih.gov/crisp/Crisp_Query.Generate_Screen •
Project Title: TECHNIQUES
DETECTING
NEONATAL
SEPSIS
USING
MOLECULAR
Principal Investigator & Institution: Jordan, Jeanne A.; Magee-Women's Health Corporation 204 Craft Ave Pittsburgh, Pa 152133180 Timing: Fiscal Year 2002; Project Start 16-AUG-2000; Project End 31-MAY-2004 Summary: (Adapted from the Investigator's Abstract): Approximately 150,000 to 500,000 infants greater than 34 weeks gestational age are admitted to NICUs annually who are at risk for systemic infection. These infants all receive systemic antibiotic therapy. However, the vast majority of these infants are not infected, but rather have symptoms that are secondary to other medical conditions. The current standard of care for evaluating neonatal sepsis is blood culturing, which lacks sensitivity and is not informative prior to at least 24 to 46 hours. As a result, there is prolonged use of antibiotic therapy in many newborns that could be shortened if a more rapid test to rule out sepsis was available. This application proposes to investigate two potential tests, a PCR-based amplification assay for detecting bacterial 16S ribosomal DNA and an ELISA-based assay for defensins, to determine their usefulness as early predictors of systemic neonatal infection. Website: http://crisp.cit.nih.gov/crisp/Crisp_Query.Generate_Screen •
Project Title: DEVELOPMENT EXPRESSION OF CHEMOKINES AND THEIR RECEPTORS Principal Investigator & Institution: Calhoun, Darlene A.; Pediatrics; University of South Florida 4202 E Fowler Ave Tampa, Fl 33620
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Sepsis
Timing: Fiscal Year 2002; Project Start 13-AUG-2001; Project End 31-JUL-2003 Summary: (provided by applicant) Despite the development of newer and more effective antibiotic therapies, sepsis related mortality in neonates undergoing intensive care has remained constant for nearly two decades. The rate of infection among these neonates ranges from 25% to 50%, with bacterial infection remaining a major cause of death and long-term morbidity. The cost of caring for premature neonates, who are the most susceptible to infection, represents over 50% of the total dollars expended for neonatal intensive care unit (NICU) services. The unique susceptibility of the human neonate to serious and overwhelming bacterial infections relates in part to deficiencies of antibody, complement, and T lymphocytes. However, while deficiencies in these contribute to the neonate s susceptibility, neutrophil defects appear to be the major host defense abnormality. Functional defects in neonatal polymorphonuclear leukocyte adherence, aggregation, chemotaxis, phagocytosis, and intracellular killing have been described in both the term and preterm infant. Of these defects, neutrophil chemotaxis, as assessed by in vitro assays, is abnormal at birth. While term infants rapidly establish normal chemotactic function, the process of postnatal maturation begins two to three weeks after birth in the preterm infant and proceeds very slowly. Neutrophils follow a concentration gradient of chemotactic factors in their movement from the vascular compartment to the site of microbial invasion. Chemokines are chemotactic cytokines that largely control leukocyte migration. While considerable information has emerged in the past ten years related to the role of chemokines in the adult, very little information exists as to the physiologic significance of chemokines in the neonate. Candidate chemokine/chemokine receptors for influencing neutrophil chemotaxis include members of the CXC chemokines. We propose that understanding the gene regulation for the expression of specific chemokine receptors (CXCR) during development would enhance our understanding of chemotaxis in the neonate. We further propose that defining circulating concentrations of chemokines critical for neutrophil migration would be essential to understanding their role in both health and disease. Website: http://crisp.cit.nih.gov/crisp/Crisp_Query.Generate_Screen •
Project Title: DEVELOPMENT OF NEW LIPID A BINDING AGENTS Principal Investigator & Institution: Savage, Paul B.; Associate Professor; Chemistry and Biochemistry; Brigham Young University A-261 Asb Provo, Ut 846021231 Timing: Fiscal Year 2002; Project Start 01-JUN-1998; Project End 31-MAY-2003 Summary: Sepsis affects the lives of hundreds of thousands of people each in the U.S. Sepsis caused by Gram-negative bacteria results from adverse host immune response to the Lipid A (LA) portion of endotoxin. Compounds with high affinity for LA, including polymyxin B (PMB) and derivatives, are capable of detoxifying LA nd protection a host against sepsis. By binding LA, these compounds also sensitize Gram-negative bacteria to hydrophobic antibiotics. However, therapeutic use of PMB is limited due to its toxicity. The aim of this research is to develop small molecules capable of strong and selective associating with LA for use in treatment of sepsis and as means of fighting bacterial infection. Taking the interactions of PMB with LA as a model, this research focuses on the preparation of simple compounds, based on cholic acid scaffolding, capable of mimicking the LA-binding behavior of PMB but lacking the toxicity of the antibiotic. Simple cholic-acid based LA binders will present many advantages over reported LA binding molecules including: ease of preparation and derivatization, greater control over biological stability, and potential oral bioavailablity. Cholic acid derivatives were designed to mimic a conformation of PMB believed to be important in its association with LA. Preliminary experiments with cholic acid derivatives
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demonstrate their ability to sensitize Gram- negative bacteria to hydrophobic antibiotics, a behavior of LA binding agents. Optimization of LA-binding characteristics will be achieved by preparing libraries of compounds made up by amino acids linked to cholic acid scaffold. The libraries will be screened for LA binding using affinity chromatography. The affinity chromatography stationary phase will be made up of LA immobilized through hydrophobic interactions of C18- silica particles or polystyrene beads. The types of amino acids in effective LA binders will be determined via mass spectroscopy. New LA-binding agents will be tested for the ability to sensitize Gramnegative bacteria to hydrophobic antibiotics and/or inhibit the effects of LA on human monocytes (specifically interleukin 1b production). Association of PMB and the new LAbinders with LA and LA derivatives will be compared using microtitration calorimetry. Website: http://crisp.cit.nih.gov/crisp/Crisp_Query.Generate_Screen •
Project Title: DIAPHRAGM MITOCHONDRIAL ALTERATIONS IN SEPSIS Principal Investigator & Institution: Callahan, Leigh A.; Medicine; University of Rochester Orpa - Rc Box 270140 Rochester, Ny 14627 Timing: Fiscal Year 2002; Project Start 01-JUN-2001; Project End 31-JUL-2002 Summary: (Applicant's abstract): Recent work suggests that mitochondria dysfunction plays a central role in sepsis, a major cause of death and morbidity in the United States. The underlying mechanisms responsible for this mitochondria dysfunction are not known. The goal of the present proposal is to test the hypothesis that increased free radical generation in sepsis produces specific biochemical, structural and genetic changes that result in marked physiologic alterations in mitochondrial function. We postulate: (a) mitochondria dysfunction in sepsis results from physiologic derangements of Krebs cycle enzymes, Complex I-IV electron transport chain components, and sarcomericcreatine kinase, (b) these physiologic changes are due, in turn, to alterations in the content and composition of mitochondrial proteins, and (c) protein changes are due, in part, to free radical-mediated decrements in mitochondrial gene transcription, expression, and translation. These hypotheses will be tested in three groups of experiments, using a model of endotoxin-induced sepsis. The purpose of Objective 1 is to fully characterize the specific physiologic derangements in the mitochondria in sepsis; we will examine Krebs cycle enzyme activities, evaluate specific performance of complexes within the electron transport chain, assess sarcomeric mitochondrial creatine kinase activity, and perform a metabolic control analysis. Objective II will identify changes in the content and composition of mitochondrial protein constituents (i.e. electron transport chain protein subunits, Krebs cycle enzymes, creatine kinase) and compare the time course of these alterations with the development of physiologic abnormalities determined in Objective I. Objective III will evaluate transcription, expression, and translation of mitochondria and nuclear genes encoding for mitochondrial proteins found to be depleted in Objective II. In all studies, we will determine the role of free radical modulation of these sepsis-induced changes. Our preliminary data provide the first evidence of substantial sepsis-associated oxidative modification and depletion of mitochondria protein subunits in Complexes I, III and IV, significant alterations in NADH generation via Krebs cycle enzymes, major decreases in mitochondria creatine kinase activity, and key free radical-mediated changes in gene expression of mitochondrial proteins in sepsis. These data suggest that the proposed experiments should provide important information regarding the pathogenesis of mitochondrial dysfunction in sepsis. Website: http://crisp.cit.nih.gov/crisp/Crisp_Query.Generate_Screen
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Sepsis
Project Title: EFFECTS OF VASOACTIVE DRUGS ON PERFUSION IN SEPTIC SHOCK Principal Investigator & Institution: Murray, Patrick T.; Anesthesia and Critical Care; University of Chicago 5801 S Ellis Ave Chicago, Il 60637 Timing: Fiscal Year 2002; Project Start 01-AUG-2001; Project End 31-JUL-2006 Summary: (provided by applicant): This Mentored Patient-Oriented Research Career Development Award application requests support (or a proposal which includes mentorship by an experienced investigator, multidisciplinary support from the Chairs and faculty of several Departments. formal training in an NIH-funded Clinical Research Training Program. and a research project dissecting the regional and systemic effects of traditional and novel vasoactive drugs in critically ill patients with septic shock. Sepsis. the systemic inflammatory response to infection. may lead to refractorv hypotension (septic shock) and multiple organ system failure (MOSF). Despite increased cardiac output and oxygen delivery, death often ensues from refractory hypotension or subsequent MOSF. Standard indicators of adequate tissue perfusion which are used to titrate therapy in hypovolemic or cardiogenic shock are unreliable in hyperdynamic septic shock. Rational septic shock therapy would preferably be guided by targeted interventions tc? optimize end-organ perfusion and function, and reverse detectable tissue hypoperfusion. The primary end-organ index of adequate perfusion used in current clinical practice is renal perfusion and function. The effects of vasoactive drug therapy on renal perfusion and function are reliably quantifiable with sophisticated existing technology. This application seeks to determine the regional circulatory effects of restoring vascular contractility with standard exogenous catecholamines and the novel use of exogenous vasopressin, alone or in combination, in septic humans, focusing on renal perfusion and function. We will also examine the systemic and regional effects of targeted vasodilator therapy with fenoldopam (a novel dopaminergic agonist) in septic humans, alone or in combination with the aforementioned vasoconstrictors. Specifically, we will address the hypotheses that: 1) Addition of inotropic support (badrenoceptor stimulation) to intravenous fluids alone, or in combination with pure vasopressor therapy (a-adrenoceptor stimulation), improves renal perfusion and function in patients with sepsis or septic shock. 2) Titration of vasopressor therapy to a mean arterial pressure above the lower renal autoregulation threshold improves renal perfusion in patients with vasopressor-dependent septic shock. 3) Vasopressin therapy restores septic vascular contractility, augments vasopressor-responsiveness, reverses hypotension. and improves systemic and renal perfusion in human subjects receiving standard therapy for sepsis or septic shock. 4) Septic renal vasoconstriction is reversed, and renal perfusion and function improved, by selective renal vasodilator therapy. The overall aim of these experiments is to develop rational pharmacologic regimens and strategies for hemodynamic support in septic shock. focusing on prevention and management of septic acute renal failure, as a surrogate endpoint to optimize systemic perfusion in hyperdynamic states. Website: http://crisp.cit.nih.gov/crisp/Crisp_Query.Generate_Screen
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Project Title: PKCDELTA
ENDOTHELIAL
BARRIER
FUNCTION
MODULATION
BY
Principal Investigator & Institution: Harrington, Elizabeth O.; Assistant Professor; Medicine; Brown University Box 1929 Providence, Ri 02912 Timing: Fiscal Year 2002; Project Start 01-AUG-2001; Project End 31-JUL-2005
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Summary: Increased microvascular permeability causes pulmonary edema in acute lung injury, adult respiratory distress syndrome (ARDS), and multisystem organ failure. Disruption of endothelial cell-cell contacts is a possible cause of increased microvascular permeability. Identification of intracellular mechanisms which regulate changes in vascular permeability may lead to therapeutic strategies for controlling tissue damage due to sepsis or other causes of acute lung injury. Our preliminary results indicate that overexpression of protein kinase Cdelta (PKCDELTA) protects against thrombininduced increases in endothelial monolayer permeability. The hypothesis of this proposal is that PKCDelta blunts changes in endothelial monolayer permeability by modulating the organization of cytoskeleton junctional complexes. Endothelial cells (EC) normally form a tightly adherent monolayer. During an inflammatory or thrombogenic response endothelial monolayer permeability increases with the formation of interendothelial cell gaps as a result of both EC contraction and disruption of endothelial cell-cell interactions. EC contraction occurs by intracellular calcium mobilization, Rho GTPase activation, myosin light chain phosphorylation, actin microfilament reorganization, and focal adhesion complex (FA) formation. Disruption of endothelial cell-cell junctions occurs concomitantly with the redistribution of the adherens junction (AJ) protein complexes at the points of intercellular gap formation. The intracellular mechanisms by which edemagenic agents promote EC contraction and FA formation and the disassembly of the AJ complexes are not completely understood. A number of inflammatory and thrombogenic agents activate PKC, increasing monolayer endothelial permeability. The overall objective of this proposal is to determine the mechanism by which PKCDELTA modulates endothelial barrier function. Using microvascular EC which stably overexpress PKCDELTA and vector control, we will determine: I) whether diminished endothelial barrier function is due to the stabilization and association of AJ or FA with PKCDELTA; II) whether delta PKCDELTA blunts endothelial barrier function by modulating EC retraction at the level of actin stress fiber formation and/or function; and III) whether PKCDELTA reduces endothelial barrier function by altering MAPK signaltransduction pathway. The elucidation of the molecular mechanisms by which PKCDELTA decreases endothelial monolayer permeability may lead to the development of therapeutic agents which are protective against endothelial barrier dysfunction. Website: http://crisp.cit.nih.gov/crisp/Crisp_Query.Generate_Screen •
Project Title: ENDOTHELIN CONVERTING ENZYME IN SEPSIS: CARDIAC FUNCTION Principal Investigator & Institution: Sharma, Avadhesh C.; Pharmaceutical Sciences; North Dakota State University Fargo, Nd 581055756 Timing: Fiscal Year 2003; Project Start 01-AUG-2003; Project End 31-JUL-2005 Summary: (provided by applicant): The long-term objective of our group is to characterize an interactive role of vasoactive substances in sepsis-induced myocardial dysfunction and related cardiovascular pathologies. Chronic peritoneal sepsis in our rat model produces myocardial dysfunction in an isolated heart preparation and cardiomyocytes. Induction of sepsis also increases susceptibility of the isolated hearts to a calcium paradox-mediated myocardial injury. In vivo, we have demonstrated that induction of sepsis results in disproportionate alterations in the circulating levels of Endothelin-1 (ET-1) and nitric oxide byproducts (nitrite and nitrate, NOx). Recently we observed that inhibition of metalloprotease (endothelin-converting enzyme [ECE], which converts proET-1 to ET-1) at the time of induction of endotoxemia decreased the expression of myocardial inducible nitric oxide synthase (iNOS) and downregulated the
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Sepsis
expression of p38 mitogen-activated protein kinase (MAPK) twenty-four hours later. Therefore, our immediate objective in the present proposal is to test the hypothesis that sepsis-induced alteration in the biosynthesis of myocardial ET-1 (regulated by ECE-1) via MAPK-dependent or -independent mechanism(s) would affect NOS proteins and cardiac function. The following two specific aims are designed to address this hypothesis. Specific Aim 1: To determine if ECE-1 inhibition at the time of induction of sepsis would affect sepsis-induced myocardial dysfunction (decrease in the rates of left ventricular contraction and relaxation, i.e., + dP/dt and -dP/ dt respectively) and the expression of p38MAPK and iNOS proteins at 12 and 24 h post sepsis (Year 1-2). Specific Aim 2: To determine if ECE-1 inhibition at the time of induction of sepsis via p38MAPK-dependent or -independent mechanism would affect myocardial function in an isolated heart preparation at 24 h post sepsis (Year 2-3). The specific aims were designed to assess if ECE-1 inhibition during sepsis would suppress sepsis-induced myocardial dysfunction characterized by downregulation of p38 MAPK and depressed expression of iNOS proteins. The novel aspect of the specific aims is that the results will provide evidence for a causal relationship between ET-1 biosynthesis and the expression of p38MAPK and iNOS proteins in the myocardium. An increased understanding of the underlying mechanisms during these two stages (12 and 24 h post sepsis) of sepsis will help design therapeutic interventions for early and late stages of sepsis. Website: http://crisp.cit.nih.gov/crisp/Crisp_Query.Generate_Screen •
Project Title: ETHANOL PROMOTES LUNG ENDOTHELIAL:NEUTROPHIL INTERACTIONS Principal Investigator & Institution: Brown, Lou Ann S.; Professor; Emory University 1784 North Decatur Road Atlanta, Ga 30322 Timing: Fiscal Year 2003; Project Start 01-FEB-2003; Project End 31-DEC-2007 Summary: (provided by applicant): Investigators in this Center Application have identified chronic alcohol abuse as a co-morbid variable that significantly increases the incidence and severity of the Acute Respiratory Distress Syndrome (ARDS). The overall theme of this Center application postulates that chronic ethanol ingestion enhances the risk of alveolar flooding in response to inflammatory mediators and activated neutrophils. This suggests a "two-hit" model wherein chronic ethanol ingestion constitutes the first hit but does not generate pulmonary dysfunction. However, the first hit does predispose the lung to an enhanced response to the inflammatory mediators and activated neutrophils produced during sepsis. We postulate that one mechanism by which chronic ethanol ingestion predisposes the lung to endothelial barrier dysfunction involves ethanol-induced decreases in the availability of the antioxidant glutathione (GSH), particularly the mitochondrial pool in pulmonary microvascular endothelial cells (MVEC). When GSH availability decreases, the reactive oxygen species (ROS) generated during normal respiration and ethanol detoxification becomes amplified. With chronic exposure to ROS, the MVEC is altered and the basal expression of adhesion molecules are upregulated. During sepsis, the adhesion of the activated neutrophils and the microenvironment is potentiated. The ethanol-induced GSH depletion then amplifies the ROS produced resulting in mitochondrial dysfunction and decreased ATP generation. With limited ATP availability, the MVEC are more susceptible to cytotoxinand neutrophil-induced apoptosis and necrosis. When MVEC death is enhanced, barrier dysfunction and neutrophil migration is potentiated. Furthermore, we propose that GSH precursors will attenuate this enhanced endothelial:neutrophil interaction and result in decreased sepsis-induced acute lung injury. Using a rat model of chronic ethanol ingestion, three Specific Aims will explore this hypothesis to determine: 1) if
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chronic ethanol ingestion potentiates inflammatory mediator-induced oxidative stress in MVEC, 2) if ethanol-induced chronic ROS up regulates MVEC adhesion factor expression and neutrophil transmigration during sepsis and 3) if intervention with GSH precursors will attenuate ethanol potentiation of adhesion factor expression and neutrophil transmigration during sepsis. This proposal will improve our understanding of the relationship between alcohol and lung injury and potentially identify strategies for ARDS prevention and treatment. Website: http://crisp.cit.nih.gov/crisp/Crisp_Query.Generate_Screen •
Project Title: FUNCTIONAL AND METABOLIC PROPERTIES OF TOXIC NEUTROPHILS Principal Investigator & Institution: Mccall, Charles E.; Professor; Internal Medicine; Wake Forest University Health Sciences Winston-Salem, Nc 27157 Timing: Fiscal Year 2002; Project Start 01-APR-1999; Project End 31-MAR-2004 Summary: Sepsis is the major cause of morbidity and mortality in critical care units in the USA. During sepsis, expression of pro-inflammatory genes is repressed when blood leukocytes are stimulated ex vivo with bacterial endotoxin. Endotoxin tolerance may reflect a state of immunosuppression that contributes to the high mortality rate observed in sepsis. The objective of this proposal is to define the molecular basis of endotoxin tolerance, using three models: blood leukocytes of patients with sepsis, blood monocytes, and THP-1 pro-monocytic cells. We will test the hypothesis that a labile factor(s) mediate the tolerant phenotype by repressing the transcription of proinflammatory genes such as IL-1beta and PGHS-2. Aim 1 using a biochemical approach and an in vitro transcription assay to identify, characterize, and purify the transcription repressor(s). An alternative genetic approach seeks to identify the transcription repressor in endotoxin tolerant THP-1 cells using differential display RT-PCR. Aim 2 investigates the intracellular signaling pathways utilized by endotoxin to modulate proinflammatory gene expression. Constitutively active or dominant-negative mutants of kinase mediators are employed in co-transfection assays to test the impact of various kinase activities on IL-1beta transcription. Immunologic approaches are used to identify activated/inactivated kinases in the normal and tolerant phenotypes. Aim 3 proposes to translate our understanding of the in vitro endotoxin tolerant THP-1 cell model to the in vivo tolerant phenotype of septic patients. A genetic approach, using differential display RT-PCR, and a biochemical approach, using the purified repressor, will compare the two phenotypes. Immunological techniques also will be applied to the septic patient model to identify kinases that regulate endotoxin responses and they be activated/inactivated in the tolerant phenotype. These investigations will increase our understanding of the mechanisms that regulate pro- inflammatory gene expression and contribute to improving the management of patients with sepsis. Website: http://crisp.cit.nih.gov/crisp/Crisp_Query.Generate_Screen
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Project Title: DISCOVERY
GBS:
EPIDEMIOLOGIC
CHARACTERIZATION
FOR
GENE
Principal Investigator & Institution: Foxman, Betsy; Professor; Epidemiology; University of Michigan at Ann Arbor 3003 South State, Room 1040 Ann Arbor, Mi 481091274 Timing: Fiscal Year 2003; Project Start 01-JAN-2003; Project End 31-DEC-2007 Summary: (provided by applicant): Group B Streptococcus (GBS) is a major cause of neonatal sepsis and meningitis. First emerging as a pathogen in newborn nurseries in the 1970s, it is now a pathogen of concern in nursing homes, intensive care units and
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Sepsis
outpatient settings. Little is known about GBS virulence factors. The overall goals of this application are 1) to use genetic fingerprinting techniques to better describe the molecular epidemiology of GBS of several epidemiologically defined collections; and 2) to identify new genes associated with GBS disease. By combining epidemiologic information with molecular genetics, we can maximize our ability to detect GBS virulence-related genes associated with invasive disease, extra-intestinal colonization, transmission and antibiotic resistance. This strategy also gives us insight as to the relative importance of the identified genes and their potential mechanism. A better understanding of the molecular epidemiology and the identification of new GBS virulence genes will facilitate the discovery of new therapies and prevention strategies for GBS disease. Website: http://crisp.cit.nih.gov/crisp/Crisp_Query.Generate_Screen •
Project Title: GENETIC AND INFLAMMATORY MARKERS OF SEPSIS Principal Investigator & Institution: Angus, Derek C.; Professor; Critical Care Medicine; University of Pittsburgh at Pittsburgh 350 Thackeray Hall Pittsburgh, Pa 15260 Timing: Fiscal Year 2002; Project Start 01-APR-2001; Project End 31-MAR-2005 Summary: (Verbatim from the Applicant's Abstract): Sepsis, a diffuse inflammatory response to infection, occurs in over 450,000 cases per year in the US and frequently progresses to organ dysfunction and death. Although experimental studies using cells and animals have greatly improved our understanding of the pathophysiology of sepsis, there remains a remarkable paucity of biochemical and genetic data regarding the natural history of this important public health problem. In particular, there is a need for better markers of sepsis and outcome and a more rigorous evaluation of the complex relationships among the many genetic, inflammatory, and clinical factors that appear to influence the development and outcome of sepsis. Because pneumonia is the most common cause of sepsis, patients with this infection represent an excellent clinical model for studying sepsis in a relatively homogeneous population. We propose to study a large cohort of patients (n=2,703) with community-acquired pneumonia (CAP). Our study will be "piggy-backed" onto a multicenter trial of alternative hospital quality improvement initiatives that is already funded and slated to begin enrolling patient's early in 2001. In addition to collecting detailed clinical data, we will carry out careful genetic analyses, focusing on allelic variations in the coding or noncoding regions of genes whose products are important in the expression and/or regulation of the inflammatory response. We will also obtain measurements over time of the plasma concentrations or cell surface expression of several key inflammatory molecules. We will determine the influence of specific polymorphisms on the development, course and outcome of pneumonia and sepsis. We will test whether genetic predisposition to an exuberant inflammatory response protects against infection yet also increases risk for adverse systemic effects and outcome. We will compare genetic data from patients with results obtained from a cohort of healthy controls (n=300). We will test several existing hypotheses regarding the association of circulating inflammatory molecules with outcome. We will use time-varying regression analyses and probabilistic networks to explore in new detail relationships among genetic polymorphisms and the inflammatory response in sepsis. Finally, we will construct and evaluate two sets of clinical decision tools: i.) clinical risk prediction rules that incorporate genetic and inflammatory response variables with existing clinical factors, and; ii.) a state-transition simulation model of the course of sepsis that allows time-dependent estimates of the effects of alternative treatment decisions. This study will generate: new and valuable information regarding existing lines of inquiry and laboratory investigation; new
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hypotheses arising from the use of time-dependent modeling; and new clinical decision tools that have immediate and practical value for designing clinical trials and improving patient care. Website: http://crisp.cit.nih.gov/crisp/Crisp_Query.Generate_Screen •
Project Title: GENETIC MUTATIONS THAT PREDOPOSE TO INFECTIOUS DISEASE Principal Investigator & Institution: Butler, Bruce; Scripps Research Institute Tpc7 La Jolla, Ca 92037 Timing: Fiscal Year 2002; Project Start 30-SEP-2002; Project End 31-JUL-2007 Summary: The innate immune response is dependent upon a discrete collection of proteins that mediate the detection of microbial invaders and their elimination. Only a handful of these proteins are known at present, and among them, the Toll-like receptors and associated signaling proteins play a prominent role as sensors. The program of which this Bridging proposal is a part will permit the identification of many critical genes involved in the innate immune response. We propose to examine each of these genes in turn, starting with the Toll-like receptors and moving to new genes as they are discovered. Our ultimate goal is to determine whether mutations at these loci contribute to susceptibility to sepsis in humans, or influence the outcome of sepsis once it is established. The approach taken is one in which modem techniques for mutation detection will be used to assay genetic load within the coding region at each locus, in disease populations and in ethnically matched control populations. In particular, we will emphasize the use of advanced methods for mutation detection, coupled with highthroughput sequencing to achieve this end. In accordance with the emerging principle that most complex human diseases may be ascribed to low-frequency codominant mutations affecting multiple loci, it is expected that mutations will be over-represented in specific loci within a disease population, given that those loci encode genes that are important in fighting infection. Preliminary work has supported this principle as it applies to the TLR4 locus in meningococcal disease: there is a significant excess ofmissense mutations at this locus in the disease population, compared with controls. We intend to generalize the principle, seeking to establish the importance of rare mutations at multiple loci in the pathogenesis of diverse infections. This proposal will entail extensive interactions between the Forward Genetics Core, the Genomics Core, and the Sequencing Core. Mutational data bearing on the involvement of specific genes in sepsis will quickly be disseminated to the scientific community at large through a webbased information system. Ultimately, these studies may foretell susceptibility to infection, and explain the strong heritability of infectious diseases in molecular terms. Website: http://crisp.cit.nih.gov/crisp/Crisp_Query.Generate_Screen
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Project Title: GLU6PASE AND 6P2K/FBASE GENE REGULATION IN SEPSIS Principal Investigator & Institution: Maitra, Subir R.; Emergency Medicine; State University New York Stony Brook Stony Brook, Ny 11794 Timing: Fiscal Year 2001; Project Start 01-AUG-1999; Project End 31-JUL-2004 Summary: Sepsis is that critical condition of the patient which, if uncorrected, leads to death. The initial hyperglycemia seen in sepsis is followed by hypoglycemia with concomitant hyperlacticacidemia. Glucose-6-phosphatase (Glu-6-Pase) is a key enzyme in the homeostatic regulation of blood glucose concentration, which catalyses the final step in gluconeogenesis and glycogenolysis. Another important enzyme for regulation of gluconeogenesis is 6-phosphofructo-2-kinase/fructose-2,6bisphosphatase
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(6PF2K/FBPase), which is responsible for the production and hydrolysis of fructose-2,6bisphosphate (Fru-2,6- P2), a modulator of glycolytic to gluconeogenic flux. We have demonstrated that gene expression of Glu-6-Pase and 6PF2K/FBPase are reciprocally modulated during hyperglycemic phase of hemorrhagic shock. In this proposal, we are focusing on the regulation of expression of these two genes as models of hormonal counter-regulation during sepsis. In addition to studying the hormonal regulation of these genes in sepsis, we also propose to determine the hormonal response elements of these genes by promoter analysis. Sepsis will be induced in fasted, anesthetized rats by cecal ligation and puncture (CLP) method and will be observed for 0.5 h, 3 h, and 20 h periods. Control rats will undergo sham operation and will be observed for the same time. The liver and kidney from control and CLP rats will be freeze-clamped in liquid nitrogen and stored at -70 degrees C for future assay of glucose-6- phosphate, fructose-6phosphate, glucose-6-phosphate dehydrogenase activity, Glu-6-Pase and 6PF2K/FBPase enzyme activity and gene expression. Blood samples will be collected at the same time periods to assay glucose, corticosterone, insulin and glucagon concentration in the plasma. Relatively selective pharmacologic antagonists of hormones and cytokines will be administered prior to CLP to observe their effects on Glu-6-Pase and 6PF2K/FBPase gene expression. Hepatocytes will be isolated from control and CLP rats and will be incubated with agonists to observe their effects on Glu-6-Pase and 6PF2K/FBPase gene expression. Freshly prepared, primary cultures of hepatocytes will be transfected and then incubated with hormones and hormonal response element will be confirmed by DNA footprinting. The results of this study are essential to understand the molecular basis of deranged metabolism in sepsis. Understanding the mechanism might be useful for developing appropriate therapeutic intervention during sepsis. Website: http://crisp.cit.nih.gov/crisp/Crisp_Query.Generate_Screen •
Project Title: GLUCOSE-6-PHOSPHATE DEHYDROGENASE DEFICIENCY AND SEPSIS Principal Investigator & Institution: Spolarics, Zoltan; Associate Professor; Cell Biology & Molec Medicine; Univ of Med/Dent Nj Newark Newark, Nj 07103 Timing: Fiscal Year 2002; Project Start 01-FEB-1997; Project End 31-JAN-2005 Summary: Glucose-6-phosphate dehydrogenase (G6PD) deficiency is the most common human genetic polymorphism. A clinically significant variant is the type A deficiency present in 10-12% of African Americans. G6PD is a major supporter of cellular redox status. Our clinical investigations on young African American trauma patients revealed that after severe trauma, the type A- G6PD deficiency predisposes to the development of bacteremic sepsis, an augmented inflammatory response and worsens anemia. The defect also alters the trauma-induced monocyte responses. We will investigate the impact of the defect on the sepsis-associated multiple organ dysfunction/failure and mortality after major trauma. We will test if a compromised RBC function and an altered activation status of the reticolo-endothelial/monoculear phagocyte system contribute to the adverse clinical effects of the deficiency. The study use G6PD deficient and nondeficient human endothelial cells (HUVEC), monocytes and neutrophils and an animal model. Hypotheses: 1: After severe trauma, the sepsis-associated mortality is greater in G6PD deficient patients than non-deficient patients. The adverse clinical course of G6PD deficient patients is associated with a diminished production of antiinflammatory cytokines by monocytes, increased hemolysis, and decreased RBC deformability. The prospective cohort study will compare the clinical parameters in G6PD deficient and nondeficient patients after major trauma (ISS>13) and the time dependent changes in cytokine patterns representative of the proinflammatory/anti-
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inflammatory balance in leukocytes. Alterations in RBC deformability, lipid peroxidation, nitrosylated protein, nitroso-thiol and glutathione content will also be followed. 2: After oxidative stress, G6PD deficient endothelial cells and monocytes display an augmented pro-inflammatory response compared to non-deficient cells. Activation of redox-dependent transcription factors ( NFkB,AP1,SP1) and redox status (GSH/GSSG) and the accompanying changes in cytokine production will be determined after ischemia- reoxygenation or chemically-induced oxidative stress in deficient and non-deficient HUVEC and monocytes. 3. G6PD deficiency diminishes the antioxidant capacity of endothelial cells more severely than the antioxidant capacity of phagocytes which results in an enhanced phagocyte-mediated endothelial dysfunction. Phagocytemediated endothelial cell apoptosis/injury will be measured in co-cultures using deficient or non-deficient cells. Using specific inhibitors of G6PD in an animal model will test endothelial cell dysfunction. The studies will elucidate if the adverse clinical effects of G6PD deficiency are manifested in elevated mortality and worsening organ dysfunction after injuries. The study will reveal important and novel information on the potential mechanisms, including RBC dysfunction, monocyte activation, and the role of redox regulated gene expression that is responsible for the adverse clinical effects of G6PD deficiency in trauma patients. Website: http://crisp.cit.nih.gov/crisp/Crisp_Query.Generate_Screen •
Project Title: THERAPEUTICS
GRAM-NEGATIVE
SEPSIS:
PHARMACOPHORE-BASED
Principal Investigator & Institution: David, Sunil A.; Molecular Biosciences; University of Kansas Lawrence Youngberg Hall Lawrence, Ks 660457563 Timing: Fiscal Year 2003; Project Start 01-AUG-2003; Project End 31-JAN-2008 Summary: (provided by applicant): Gram-negative sepsis, a common and serious sequel of systemic bacterial infections is the leading cause of mortality, accounting for some 200,000 fatalities annually in the US alone. The pathogenesis of Gram-negative septic shock is due to the host response to endotoxins, or lipopolysaccharides (LPS), present on the surface of gram-negative bacteria. There are, to date, no FDA-approved therapeutic options targeting the endotoxin itself to prevent or treat this disease. We have shown that relatively simple, and synthetically easily accessible molecules of the lipopolyamine class specifically bind to LPS and neutralize its toxicity both in vitro and in animal models of septic shock. The affinity of the lipopolyamines toward LPS, however, is relatively weak (2-10 (M). In this proposal, our goal is to identify highaffinity LPS binders with nonlipopolyamine scaffolds as novel leads for the therapy of Gram-negative sepsis. A focused library of ~6000 compounds, each possessing the primary pharmacophore for LPS binding will be screened using a well-established fluorescence displacement method implemented in HTS formats. Binding, however, does not necessarily manifest in neutralization of LPS toxicity. For neutralization, an additional, appropriately positioned long-chain aliphatic group is essential. Highaffinity binders ("hits") identified in HTS will be alkylated appropriately to generate LPS-neutralizing compounds (sequestrants). In in vitro assays, the potency of lead compounds in inhibiting the release of LPS-mediated proinflammatory cytokines such as tumor necrosis factor will be characterized. In a select subset of promising leads identified in the screens described above, we will verify that the mechanism of action of inhibition of LPS toxicity is via its sequestration by showing that relevant upstream cellsignaling events are blocked. The protective effects of particularly promising molecules will then be examined in two well-established murine models of gram-negative sepsis. We will systematically evaluate the toxicity of the test-compounds in a carefully chosen
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panel of in vitro assays. Molecular modeling techniques will be applied in an effort to correlate experimentally observed binding affinities of the test compounds with features of molecular interaction such as binding geometry, H-bonds, electrostatic, hydrophobic, and van der Waals contributions to the free energy of binding. Based on the data from the primary screen, in silico modeling, and biological assays, we will synthesize a series of analogues around promising leads using a combination of focused virtual library screening and classical medicinal chemistry approaches. Website: http://crisp.cit.nih.gov/crisp/Crisp_Query.Generate_Screen •
Project Title: HEME OXYGENASE-1 AND SEPSIS USING TRANSGENIC MICE Principal Investigator & Institution: Choi, Augustine M.; Professor; Medicine; University of Pittsburgh at Pittsburgh 350 Thackeray Hall Pittsburgh, Pa 15260 Timing: Fiscal Year 2002; Project Start 01-JAN-1998; Project End 31-MAY-2003 Summary: (Adapted from the applicant's abstract): Heme oxygenase (HO) catalyzes the first and rate-limiting step in the oxidative degradation of heme to bilirubin. While HO2 is constitutively expressed, HO-1 is highly induced by heme, metal ions, cytokines, and agents causing oxidative stress such as LPS during gram negative sepsis. The PI's laboratory has shown that HO-1 induction may play a role in protection against oxidative stress in an in vivo model of septic shock and MOSF. In this proposal the investigators propose to utilize HO-1 transgenic and knockout mice to test their hypothesis that HO-1 plays a critical role in providing protection against oxidative stress. Specifically, they propose to 1) generate and identify transgenic mice overexpressing HO-1 selectively in the endothelial and vascular smooth muscle cells in the vascular wall, 2) determine the physiological, biochemical and cellular responses of HO-1 overexpressing transgenic mice in a murine model of septic shock and MOSF, 3) apply HO-1 knockout mice to this sepsis model, and 4) determine the mechanism(s) by which HO-1 induction protects against oxidative stress. Website: http://crisp.cit.nih.gov/crisp/Crisp_Query.Generate_Screen
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Project Title: IL-6 AND HEPATIC DYSFUNCTION IN SEPSIS Principal Investigator & Institution: Deutschman, Clifford S.; Professor; Anesthesia; University of Pennsylvania 3451 Walnut Street Philadelphia, Pa 19104 Timing: Fiscal Year 2002; Project Start 01-JUN-2000; Project End 31-MAY-2005 Summary: ( Applicant's abstract.) Sepsis and the related Systemic Inflammatory Response Syndrome (SIRS) and Multiple Organ Dysfunction Syndrome (MODS) are important causes of death and disability in surgical or injured patients, although the exact cause of death is often obscure. These disorders are complex, involve a number of molecular mediators and effect most organs. Little is understood, however, about the manner in which organ dysfunction develops in these diseases. One important organ that is damaged in sepsis/SIRS/MODS is the lever. Intra-abdominal fecal contamination causes SIRS/MODS-like abnormalities in the liver of rodents. In this study we will examine one proposed mechanism that we believe contributes to the development of liver dysfunction in sepsis /SIRS/MODS. We have shown that intra-abdominal fecal contamination causes an early down regulation of transcription in this organ. This change affects many genes, including some encoding molecules that 1) transfer bile acids out of liver cells and into the biliary system and 2) allow liver cells to burn fat. We propose that the inflammatory cytokine IL-6 mediates some aspects of decreased gene expression, leading to liver cells that cannot excrete bile salts or burn fat. A build-up of bile salts and fat in liver cells "poisons" them so that they die. When enough liver cells
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die, liver dysfunction develops. The role played by IL-6 in this proposed mechanism of hepatic dysfunction will be studied in the setting of normal IL-6 levels, IL-6 absence, IL6 excess and IL-6 repletion after depletion. Several specific measures will be studied. These include 1) transcription of the bile acid transporters Ntcp and Mrp2 and the ratelimiting enzyme in fat oxidation, CPTII, to be determined using transcription elongation analysis, 2) activation of two hepatic nuclear proteins, C/EBPalpha and HNF-1alpha, that modulate transcription of Ntcp, Mrp2 and CPTII and 3) the development of cholestasis (bile trapping in cells) and steatosis (fat trapping in cells) as indicated by microscopic examination of fixed liver sections. In addition, we will mimic IL-6 levels in sepsis in normal mice by 1) administering intravenous IL-6 and 2) injecting a virus that is taken up by the liver and produces high intrahepatic levels of IL-6. We will then study transcription, transcription factor activation, cholestasis and steatosis. These studies should provide key information on the role played by an important inflammatory mediator, IL-6, in the complex series of events that results in the hepatic dysfunction of SIRS/MODS. Website: http://crisp.cit.nih.gov/crisp/Crisp_Query.Generate_Screen •
Project Title: IMMUNITY TO PNEUMOCOCCAL SURFACE PROTEIN A AND C Principal Investigator & Institution: Briles, David E.; Professor; Microbiology; University of Alabama at Birmingham Uab Station Birmingham, Al 35294 Timing: Fiscal Year 2002; Project Start 01-AUG-1984; Project End 31-OCT-2002 Summary: (Adapted from the applicant's abstract): The proposal is aimed at continuing Dr. Briles' research efforts in the study of pneumococcal surface antigens. The application focuses on the first cell-associated protein described, PspA, as well as a second newly discovered protein, PspC. These two proteins may serve as vaccines themselves or as protein carriers for capsular polysaccharide-protein conjugates. PspA is present on all pneumococcal strains and can elicit protective immunity against sepsis and nasopharyngeal carriage in mice. PspC is related to PspA, but larger in size, and shows virtual identity with PspA in its C-terminal half. The proposed studies will determine whether PspC is a virulence factor and whether it can elicit protection. The relative roles of PspA and PspC in virulence and carriage in nonimmune animals will be examined. In addition, the relative roles of immunity against PspA and PspC in carriage, sepsis, and spread of pneumococci from the nasopharynx will be explored. The ability of human antibody to these molecules to protect mice from infection will be evaluated. Cross-reactive regions between the PspA and PspC proteins will be identified as well as the regions of each molecule most useful as a vaccine. Immunity to PspA and PspC will be evaluated to determine whether it involves opsonization, blocks virulence functions, or acts by other mechanisms. The data obtained will assist with the development of correlates of protective immunity for PspA and PspC that can be applied to vaccine development. Website: http://crisp.cit.nih.gov/crisp/Crisp_Query.Generate_Screen
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Project Title: IMMUNOLOGICAL ASPECTS OF HEMORRHAGE Principal Investigator & Institution: Chaudry, Irshad H.; Professor, Professor, Vice Chairman & Di; Surgery; University of Alabama at Birmingham Uab Station Birmingham, Al 35294 Timing: Fiscal Year 2002; Project Start 01-APR-1988; Project End 31-MAR-2003 Summary: Our recent studies indicate that proestrus female mice [with cycle- increased levels of estrogen and prolactin (PRL)] have improved immune responses after trauma-
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Sepsis
hemorrhage as opposed to markedly depressed responses in males. Moreover, the survival rate of proestrus females following sepsis after trauma-hemorrhage was significantly higher than age-matched males. In contrast to young proestrus mice, aged females (defined by lowered estradiol levels) show marked immunosuppression after trauma. Our hypothesis, therefore, is that it is the high estradiol or a high estradiol: androgen ratio which directly (receptor- mediated) or indirectly (receptor-independent) enhance immune functions in proestrus females, and the loss of these estrogenic effects may contribute to the failure to maintain immune responses in aged females after trauma-hemorrhage. Studies are proposed to determine the mechanism of regulation of estradiol and estrone by hypothalamic/pituitary factors adrenals and aromatase activity and determine how differences in estradiol levels or the estradiol: androgen ratio due to the estrus cycle, ovariectomy (OVX, in middle aged mice to reduce estrogen), and age affect immune responses after trauma. Sex steroids (SS) receptor- mediated and receptor-independent gene activation mechanisms will be studied in T-cells and macrophages (Mphi). Since activation of AF-1 and AF-2 regions of estrogen receptor (ER) is critical for agonist and antagonist effects, activation of the ER agonist regions by estrogens in T-lymphocytes will be evaluated by transfection studies. Moreover, since SS non-ligand response also involve [Ca2+]i mobilization, T cells and Mphi will be examined for Ca2+ signal transduction and the expression and translocation of PKC isoforms. The release of TH1 and TH2 cytokines and IL-6 and the effects of PRL on their release in proestrus, OVX, aged, ER-, and PRL-knockout mice will also be evaluated. Additionally, the effect of SS on PRL and TH1 and TH2 cytokine-induced JAK-STAT expressions will be evaluated. Analysis of bone marrow for lymphoblastoid/myeloblastoid cell composition, and the effect of SS on the population of these cells will be determined. We will evaluate if administration of beta-estradiol, Raloxifene or PRL in vivo after trauma-hemorrhage improves the depressed immune responses in estrogen deficient mice. If a single dose is ineffective, multiple doses of these drugs with or without gonadotropin releasing hormone (GnRH) or flutamide (androgen receptor antagonist) will be administered to determine whether synergistic beneficial effects on immune responses are produced and whether the susceptibility to sepsis after trauma is decreased. Detailed mechanistic studies of T cell and Mphi functions using molecular biological techniques to determine why low estradiol fails to maintain immune functions in aged females after trauma and the use of estradiol, Raloxifene, PRL, GnRH or flutamide to restore immune functions should yield novel information and provide an innovative approach for improving the immune responses and reducing mortality from sepsis following trauma-hemorrhage in postmenopausal as well as in surgically OVX patients with low estrogen activity. Website: http://crisp.cit.nih.gov/crisp/Crisp_Query.Generate_Screen •
Project Title: IMPROVED LIVER FUNCTION AND REGENERATION WITH A20 Principal Investigator & Institution: Ferran, Christiane; Beth Israel Deaconess Medical Center St 1005 Boston, Ma 02215 Timing: Fiscal Year 2003; Project Start 01-JAN-2003; Project End 31-DEC-2007 Summary: (provided by applicant): Necrosis and apoptosis of hepatocytes are critical pathologic features associated with liver injury. Hepatocyte apoptosis is a feature of viral hepatitis, ischemic liver injury, sepsis, cholestasis, and a result of exposure to hepatotoxic substances such as ethanol, acetaminophen and cytostatic drugs. Massive hepatocyte apoptosis and necrosis result in fulminant hepatic failure (FHF). Only 14% of patients diagnosed with FHF recover with medical therapy. Orthotopic liver transplantation (OLT) has dramatically improved the fate of these patients (49%
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undergo OLT), yet 37% die while awaiting OLT. This gloomy picture is balanced by the unique capacity of the liver to regenerate. Hepatocyte replication leads to a full recovery of liver function and mass 1-2 weeks following surgical, viral or chemical hepatic loss. We propose that protecting hepatocytes from apoptosis and promoting their proliferation are two strategies that could beneficially impact FHF. Our preliminary data demonstrate that A20 promotes hepatocyte proliferation and is anti-apoptotic. A20 is part of the physiologic response of hepatocytes to injury. A20 is upregulated in hepatocytes by pro-inflammatory stimuli including TNF and LPS and functions to protect from TNF mediated apoptosis. Gene transfer of A20 to mice livers protects from lethality in the galactosamine and LPS (D-gal/LPS) model of toxic FHF. Adenovirus mediated expression of A20 in livers of BALB/c mice yields an 89% survival rate following administration of D-gal/LPS as compared to 15-20% in control mice. Mice expressing A20 maintain normal liver function as assessed by prothrombin time while controls suffer from a severe bleeding diathesis. Expression of A20 in the liver protects from lethality associated with a subtotal (87%) liver resection (LR). In this model, resection of 87% of the liver mass results in 100% lethality. In contrast, >60% of mice expressing A20 survive the 87% LR and demonstrate increased regenerative capacity as assessed by the number of PCNA (proliferating cell nuclear antigen) positive nuclei in the liver. These results qualify A20 as a critical gene involved in accelerating liver regeneration and promoting hepatocyte survival and function, even when facing extreme metabolic demands. These encouraging results prompted the submission of this proposal. Our specific aims are i) to dissect, in vitro, the molecular basis of the (1) antiapoptotic and (2) pro-proliferative function of A20 in hepatocytes and ii) to confirm that liver directed gene therapy using A20 will beneficially impact upon toxic, FAS-mediated and surgical experimental models of FHF. From a basic science standpoint, the in vitro work proposed will address the effect of A20 upon transcription factors and expression of genes involved in apoptosis, activation and proliferation of hepatocytes. This should unveil many unknowns in our understanding of hepatocyte biology and could lead to the discovery of novel therapeutic targets. From a therapeutic standpoint, validation of the beneficial effect of A20 in the murine in vivo models of FHF should set the basis for extending this approach to models of FHF in non human primates and potentially to clinical applications. The generation of novel safer and tissue specific viral vectors for gene transfer and the development of non-viral means of protein delivery to cells will facilitate clinical translation of A20 based therapies. Website: http://crisp.cit.nih.gov/crisp/Crisp_Query.Generate_Screen •
Project Title: INVESTIGATION OF XRCC5 MUTANT MICE Principal Investigator & Institution: Hasty, Edward P.; Molecular Medicine; University of Texas Hlth Sci Ctr San Ant 7703 Floyd Curl Dr San Antonio, Tx 78229 Timing: Fiscal Year 2002; Project Start 07-JUL-1997; Project End 31-MAR-2006 Summary: Genomic instability is a major cause of cancer and believed to be a major contributor to aging. The gene Xrcc5, that codes for the protein, Ku80 is important for maintaining genomic stability by repairing double-strand breaks (DSBs) in DNA and by capping chromosomal ends. Mice deleted for Ku80 are immunodeficient due to defective repair of DSBs that occur during Variable (Diversity) Joining recombination and cells derived from ku80-mutant mice are hypersensitive to ionizing radiation and reactive oxygen species. In addition, these cells exhibit cytogenetic aberrations including telomeric fusions. ku80-mutant mice exhibit an early onset of age-related changes in a variety of tissues that are also observed in control mice; thus, Ku80 may be important for the normal aging process. These changes include growth plate closure, osteopenia, skin
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Sepsis
and follicular atrophy, degenerative processes in the liver and shortened life span. Early mortality was at least partly due to early onset of the age - specific diseases, sepsis and cancer. Even though onset of cancer is early for ku80-mutant mice, the total incidence is low. Interestingly, deletion of the tumor suppressor protein, p53, greatly increased the risk of lymphoma suggesting that Ku80 is also a tumor suppressor. The biological role of Ku80 in maintaining genomic stability will be determined during aging and oncogenesis and completion of this proposal will significantly impact our understanding of Ku80's role during these events. l: Determine the contribution genomic instability, induced by either oxidative damage or telomere dysfunction, has on the ku80-mutant phenotype. The onset and spectra of genetic mutations will be compared between ku80-mutant and control mice. To determine the impact of oxidative damage, DNA lesions will be measured in ku80- mutant mice after exposure to ionizing radiation and by overexpressing proteins that eliminate oxygen radicals, catalase and Cu/Zn-superoxide dismutase. To investigate the impact of telomere maintenance, ku80-/- mTR-/- mice will be investigated. 2: Determine the impact general genomic instability has on oncogenesis in ku80-mutant mice that are deleted for p53 in nonlymphoid tissue. The role of the tumor suppression protein, p53, will be analyzed for its impact on oncogenesis in ku80-mutant mice with specific attention given to mammary tissue in a defined mouse model. Website: http://crisp.cit.nih.gov/crisp/Crisp_Query.Generate_Screen •
Project Title: LIVER/LUNG INTERACTIONS DURING GRAM NEGATIVE ENDOTOXEMIA Principal Investigator & Institution: Matuschak, George M.; Professor; Internal Medicine; St. Louis University St. Louis, Mo 63110 Timing: Fiscal Year 2002; Project Start 01-DEC-1989; Project End 31-JUL-2003 Summary: (Adapted from the applicant's abstract): During gram-negative bacteremic sepsis, a liver-lung axis of inflammation predisposes to critical organ injury due to an imbalanced expression of inflammatory vs. anti- inflammatory gene products. Such postbacteremic organ dysfunction is thought to be augmented by secondary ischemichypoxic stress owing to reduction-oxidation (redox)-sensitive transcription factors and subsequent amplification of inflammatory responses mediated by expression of key cytokine and noncytokine genes. The objective of the proposed research is to test the hypothesis that postbacteremic O2 limitation within the liver and the lungs differentially modulates the activation of a defined group of redox-sensitive transcription factors, thereby altering cytokine expression in a directionally- opposite and organ-specific manner. Experiments are designed to determine the effects of secondary reductions in the hepatic vs. pulmonary O2 supply in modulating postbacteremic transactivation of nuclear factor-kB (NF-kB), activator protein (AP)-1, NFIL-6, and the cyclic AMP response element binding protein (CREB). The biologic significance of these changes will be assessed by examining the concomitant expression of inflammatory cytokines (TNF-alpha, IL-1alpha, IL-1beta), anti-inflammatory cytokines (IL-6, IL-10), prostaglandin (PG) H synthase-2 (COX-2) and inducible nitric oxide synthase (iNOS) in these organ systems in relation to redox status during ex situ organ perfusion. Experiments are also designed to define postbacteremic protein:DNA interactions during hypoxic stress and reoxygenation by assessing the activation of these transcription factors in Kupffer cells and alveolar macrophages. Resulting data will establish if an autoregulatory loop involving enhanced COX-2 and CREB activity suppresses postbacteremic cytokine expression by a PGE2-dependent mechanism in an organ-specific manner. The role of activation of the nuclear protein hypoxia inducible
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factor-1 in co-modulating endotoxin-induced-cytokine gene expression during subsequent hypoxia will be assessed. These endpoints will also be analyzed in conscious rats during hypoxic stress, with and without preexisting liver dysfunction following bacteremic infection. In parallel studies, the cis-acting DNA sequences that confer hypoxic suppressibility of endotoxin-induced cytokine promoter activity in RAW 264.7 cells transfected with TNF-alpha and IL-1beta reporter gene constructs will be identified. Results from these vertically-oriented studies should provide novel insights into the transcriptional regulation of cytokine and iNOS expression during gramnegative bacteremic sepsis while identifying mechanistic approaches to ameliorate lung injury and multiple organ failure. Website: http://crisp.cit.nih.gov/crisp/Crisp_Query.Generate_Screen •
Project Title: MACROPHAGE DEATH AND SEPSIS Principal Investigator & Institution: Han, Jiahuai; Scripps Research Institute Tpc7 La Jolla, Ca 92037 Timing: Fiscal Year 2003; Project Start 01-JAN-2003; Project End 31-DEC-2006 Summary: (provided by applicant): Overproduction of proinflammatory cytokines by macrophages and other cells is critical in the development of septic shock. On the other hand, immunocompromise of macrophages and lymphocytes that developed during the onset of sepsis was also believed to contribute to the lethality of this disease. The lifespan of macrophages and lymphocytes was modulated in sepsis and is responsible, in part, for the uncontrolled inflammatory response and immunodepression. Recent studies have shown that inhibition of lymphocyte apoptosis increased the survival rate of sepsis in an animal model. However, the effect of macrophage apoptosis on the outcome of sepsis has not been addressed. We found that macrophage apoptosis was induced in the same animal model when caspase inhibitor was administered to prevent lymphocyte death. We also found that Nur77 was induced in apoptotic macrophages and Nur77 induction is required for macrophage death. As reported by others using other types of cells, Nur77 induction requires transactivation of MEF2, but a signaling triggered by bacterial components is also required for macrophage expression of Nur77. This proposal will use the knowledge we have of macrophage apoptosis to promote and inhibit macrophage death in septic mice and thereby determine whether macrophage apoptosis positively or negatively affects the outcome in septic mice. We will also further elucidate the molecular mechanisms underlying the macrophage apoptosis. The proposed study will provide information regarding whether macrophage death should be avoided or enhanced in the treatment of sepsis. Our study will also lead to a better understanding of how macrophage apoptosis is controlled, which is needed to develop therapeutically useful strategies to selectively promote or inhibit macrophage death. Website: http://crisp.cit.nih.gov/crisp/Crisp_Query.Generate_Screen
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Project Title: MECHANISMS OF CELL INJURY IN BURN COMPLICATED BY SEPSIS Principal Investigator & Institution: Horton, Jureta W.; Professor; Surgery; University of Texas Sw Med Ctr/Dallas Dallas, Tx 753909105 Timing: Fiscal Year 2002; Project Start 01-AUG-1999; Project End 31-JUL-2003 Summary: (adapted from applicant's abstract): Despite aggressive fluid resuscitation and topical antimicrobial therapy after burn trauma, sepsis frequently results from the loss of dermis; thus sepsis and resultant multiorgan failure are a major cause of death in the burn unit. Studies from the PI's lab and others' have shown that burn trauma and
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sepsis independently alter cardiocirculatory performance, and recent studies suggest that myocardial abnormalities after burn, trauma or sepsis are related to intracellular accumulation of calcium with subsequent cellular injury and dysfunction. Although this field has grown rapidly, much is still unknown about the cellular mechanisms underlying cardiac dysfunction after either trauma or sepsis. The PI's group have focused their attention on a clinically relevant model of burn injury complicated by sepsis (intratracheal administration of S. pneumoniae administered 24 hours postburn) and have shown progressive cardiocirculatory dysfunction in this two-hit model. Specific Aim 1a will determine if burn/sepsis exacerbates the increased [Ca2+] and [Na2+] shown to occur after burn alone and will determine the contribution of altered Na+/Ca2+ to cardiac contractile dysfunction. Specific aim 1b will determine the contribution of transient cellular acidosis and altered H+/Na+ exchange to increased [Na2+], and whether increased [Na+] in turn promotes Na+/Ca2+ exchange in [Ca2+] overload. Specific Aim 2 will determine the contribution of burn/ sepsis-mediated alterations in SR Ca2+ handling (SR Ca2+ efflux, Ca2+-ATPase activity, SERCA, and SR Ca content) to cellular Ca2+ and cardiac contractile deficits and determine the contribution of burn/sepsis induced myofilament Ca2+ insensitivity to cardiac contractile dysfunction. Studies in Specific Aim 3 will examine the role of PKC activation in intracellular Na+/Ca2+ accumulation and cardiac contractile dysfunction in burn sepsis. Studies in Specific Aim 4 will determine the contribution of increased [Ca2+] and reactive oxygen species to apoptosis in burn/sepsis and further determine the contribution of apoptosis to burn/sepsis-induced ionic derangements as well as cardiac contractile dysfunction. Only by understanding the cellular events involved in the postburn inflammatory cascade can adequate prevention and treatment modalities be designed to improve outcome. Website: http://crisp.cit.nih.gov/crisp/Crisp_Query.Generate_Screen •
Project Title: MECHANISMS OF GROWTH HORMONE RESISTANCE IN SEPSIS Principal Investigator & Institution: Cooney, Robert N.; Staff Physician; Surgery; Pennsylvania State Univ Hershey Med Ctr 500 University Drive Hershey, Pa 170332390 Timing: Fiscal Year 2003; Project Start 01-MAY-1997; Project End 31-MAR-2007 Summary: (provided by applicant): The catabolism of protein after injury or infection results in multiple complications which prolong recovery and cause death. Nutrient intake is unable to prevent protein catabolism suggesting other factors are important. Growth hormone (GH) induces circulating insulin-like growth factor-I (IGF-I) synthesis by liver, which stimulates muscle protein synthesis. During sepsis, a 2-4 fold increase in circulating GH is seen with a 50% decrease in plasma IGF-I, a 40% reduction in muscle protein synthesis, and decreased muscle mass. The onset of GH resistance and loss of muscle in sepsis is mediated by the inflammatory cytokines, TNF and IL-I. Treating septic rats with IL-1 or TNF antagonists ameliorates the effects of sepsis on plasma IGF-I levels and muscle catabolism. The liver is the major source of circulating IGF-I, and will be the focus of this project to elucidate the mechanisms responsible for GH resistance. Postreceptor defects in GH signaling represent the predominant mechanism for GH resistance in sepsis. The magnitude of the IGF-I response to GH is determined by three distinct processes: the activation/propagation of JAK/STAT and MAP kinase signaling by GH, the regulation of IGF-I gene expression, and the termination of GH signaling. To investigate the mechanisms responsible for GH resistance, we developed a hepatocyte model of cytokine-mediated GH resistance. This novel hepatocyte model uniquely positions us to delineate the mechanisms responsible for defective GH signaling and IGF-I expression. TNF inhibits both the activation and termination of GH signaling by
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the JAK/STAT and MAP kinase pathways. IL-1 also inhibits the induction of IGF-I by GH, but the time course differs from TNF, potentially involving different mechanisms. The mechanisms responsible for GH resistance will be determined with in vitro studies involving TNF and IL-1, and in vivo studies in our rat model of sepsis. Our hypothesis is that cytokine-rnediated alterations in GH signaling mediate hepatic GH resistance and muscle catabolism during sepsis. The specific aims are: (1) to delineate the effects of sepsis, TNF and IL-1 on the activation/propagation of GH signaling; (2) to elucidate the mechanisms by which sepsis, TNF and IL-1 regulate IGF-I gene expression; and (3) to determine the mechanisms by which sepsis, TNF and IL-1 terminate GH signaling. An understanding of how cytokines and growth factors regulate protein catabolism is important for the care of septic patients. Website: http://crisp.cit.nih.gov/crisp/Crisp_Query.Generate_Screen •
Project Title: MECHANISMS OF NFKB ACTIVATION IN HYPOXIA AND SEPSIS Principal Investigator & Institution: Chandel, Navdeep S.; Assistant Professor of Medicine; Medicine; Northwestern University 633 Clark Street Evanston, Il 60208 Timing: Fiscal Year 2002; Project Start 10-FEB-2000; Project End 31-JAN-2005 Summary: The development of sepsis in critically ill patients is an ominous event that frequently leads to multiple organ failure and death. Increased release of cytokines has been implicated in the pathophysiology of these events, although the mechanisms underlying this relationship are not fully understood. Tissue hypoxia may also develop during sepsis, due to microvascular dysfunction and a failure to distribute capillary blood flow in accordance with tissue O2 need. A central hypothesis of this application is that tissue hypoxia will amplify the intracellular response to sepsis, by activating signaling pathways that regulate gene expression and subsequent release of cytokines and other inflammatory mediators. In sepsis, endotoxin (LPS) released from gramnegative bacteria complexes with binding proteins, which then bind to the CD14 receptor on cells. This initiates an intracellular signaling cascade leading to the activation of transcription factors that stimulate expression of genes including the cytokine TNFalpha and the inducible isoform of nitric oxide synthase (iNOS). TNFalpha can amplify the inflammatory cascade and may contribute to the pathophysiological state, while iNOS can result in unregulated release of nitric oxide, which may contribute to vascular dysfunction, organ failure and cell death. Activation of the transcription factor nuclear regulatory factor kappa B (NF-kappaB) is an important event in the expression of TNFalpah and iNOS. Previous studies demonstrate that cellular hypoxia (PO2 less than 40 torr) elicits an increase in release of reactive oxygen species (ROS) from mitochondria, which then activate transcription factors such as Hypoxia Inducible Factor-1 (HIF-1). This project will test the hypothesis that these ROS produced during hypoxia amplify the response to sepsis by independently activating NF-kappaB, thereby augmenting the TNFa and iNOS expression. Hypoxia may also heighten the cytotoxic effects of TNFa and NO, by augmenting oxidant stress and by contributing to the dissipation of mitochondrial potential. Proposed studies will clarify the role of mitochondrial ROS in amplifying NF-kappaB activation and TNFalpha and iNOS mRNA expression in a cellular model of sepsis. Additional studies will determine the significance of these events for cell death pathways in that model. Collectively, this work will shed new light on the intracellular signaling events during hypoxia and sepsis involving ROS, NF-kappaB and expression of TNFalpha and iNOS. Website: http://crisp.cit.nih.gov/crisp/Crisp_Query.Generate_Screen
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Project Title: METALLOPROTEINASES IN VENTILATOR-INDUCED LUNG INJURY Principal Investigator & Institution: Foda, Hussein D.; Medicine; State University New York Stony Brook Stony Brook, Ny 11794 Timing: Fiscal Year 2002; Project Start 01-APR-2000; Project End 31-MAR-2004 Summary: Acute, diffuse lung injury often complicates sepsis, gastric acid aspiration, extensive trauma, drug overdose, and other conditions. Clinically, this condition is known as the Adult Respiratory Distress Syndrome (ARDS) that is characterized by catastrophic respiratory failure requiring mechanical ventilation (MV). Although mechanical ventilation is a main stay in managing patients with this condition, several MV-associated complications have been recognized. One of the most serious potential complications of MV is the newly recognized ventilator-induced acute lung injury (VILI). The NIH NHLBI ARDS clinical trials network has just reported that MV strategies employing low tidal volume ventilation reduced mortality in ARDS by 25 percent and lead to a decrease in lung injury and multisystem organ failure. The mechanism by which high volume ventilation causes increased mortality and lung injury is not well understood. In this proposal we will pursue very exciting preliminary results suggesting that matrix metalloproteinases (MMPs) especially gelatinases are responsible for ventilator-induced lung injury in animal lungs subjected to high tidal volume mechanical ventilation. We will also examine the mechanism by which MMPs are up regulated in this type of acute lung injury. Our hypothesis is that Ventilatorinduced lung injury is caused by the release and activation of MMPs especially gelatinases. This increase in MMPs is regulated by the cytokine Extracellular matrix metalloproteinase inducer (EMMPRIN) and is caused by the exposure of lung cells to mechanical stress. To prove this hypothesis we have set several specific aims: 1) To systematically examine the induction, release and activation of MMPs especially gelatinases in an experimental rat model of ventilator-induced lung injury (VILI); 2) To examine if MMP inhibitors can prevent ventilator-induced lung injury in rat lungs; 3) To characterize the role of EMMPRIN in VILI; 4) To investigate the influence of mechanical stress as a mechanism of the increased MMP production. This research has significant implications for the management of patients with acute, severe lung injury requiring mechanical ventilation. By better understanding the mechanisms that lead to ventilatorinduced lung injury, therapeutic strategies may be devised to improve the outcome in this highly fatal condition. Website: http://crisp.cit.nih.gov/crisp/Crisp_Query.Generate_Screen
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Project Title: MICROVASCULAR ABNORMALITIES IN SEPSIS Principal Investigator & Institution: Hollenberg, Steven M.; Rush University Medical Center Chicago, Il 60612 Timing: Fiscal Year 2002; Project Start 01-MAY-1998; Project End 31-JUL-2002 Summary: Septic shock, the leading cause of death in intensive care units, is characterized by vasodilation with decreased peripheral vascular resistance, which is often refractory to exogenously administered vasopressor agents. The most important determinant of peripheral vascular resistance is the tone of resistance arterioles, and modulation of tone in these arterioles results from a complex interplay of local vasodilators and vasoconstrictors. The mechanisms involved in the refractory vasodilation seen in sepsis have not been fully elucidated. The current proposal would be the first study to investigate microvascular abnormalities in a clinically relevant model of sepsis by testing responses of resistance arterioles to a range of endogenous
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vasoactive substances. The long-term objective of this project is to elucidate the pathophysiology of the abnormalities in vascular tone seen in patients with septic shock. The underlying hypothesis is that hypotension and abnormal distribution of blood flow in sepsis result from derangements in microvascular responses to endogenous vasoactive substances. The specific hypothesis is that responses of resistance arterioles in cremaster muscles of septic rats measured using in vivo videomicroscopy will differ from controls, and that elucidation of the mechanisms of differences in vasopressor responsiveness will aid in our understanding of important pathogenetic pathways and in the development of innovative therapies for septic shock. Specific aims: 1. To test the hypothesis that a general abnormality of microvascular reactivity is present in sepsis by comparing arteriolar responses to endogenous vasopressors in septic and control animals. 2. To evaluate potential effector mechanisms of sepsis-induced vascular hyporesponsiveness by measuring the effects of inhibitors of second messenger pathways. 3. To elucidate interactions between endogenous vasopressors and vasodilators in mediating vascular hyporesponsiveness in sepsis by testing the effects of nitric oxide synthase, cyclooxygenase, and lipoxgenase inhibitors on vasopressor-induced arteriolar constriction in septic animals. 4. To test the hypothesis that overproduction of nitric oxide by cytokine- inducible nitric oxide synthase plays a pivotal role in inducing vascular hyporesponsiveness in sepsis, first by comparing the effects of selective and nonselective nitric oxide synthase inhibitors on vasopressor-induced arteriolar constriction in septic animals, and then by measuring vascular responsiveness in transgenic septic animal deficient in inducible nitric oxide synthase. Website: http://crisp.cit.nih.gov/crisp/Crisp_Query.Generate_Screen •
Project Title: MIF AND THE HOST RESPONSE TO INFECTION Principal Investigator & Institution: Bucala, Richard J.; Professor; Internal Medicine; Yale University 47 College Street, Suite 203 New Haven, Ct 065208047 Timing: Fiscal Year 2003; Project Start 01-JUN-1998; Project End 31-JAN-2008 Summary: (provided by applicant): Overwhelming infection can lead to massive endorgan damage and death, which is a clinical condition known as septic shock. Current concepts of septic shock pathogenesis emphasize the role of innate immunity and excessive inflammatory cytokine production. Under normal circumstances, these responses are carefully regulated so that the infectious agent is eliminated without causing life-threatening tissue damage. Inhibition of inflammatory cytokines has been studied as a way to treat septic shock, however therapies directed at mediators such as TNF alpha and IL-I have not shown clinical benefit, and in fact have worsened outcome in some studies. We have discovered that macrophage migration inhibitory factor (MIF) is an important mediator of innate immunity and upstream regulator of inflammatory cytokine production. Anti-MIF protects from shock caused by live bacteria in the same models of infection that have demonstrated either a null or a detrimental effect of antiTNF alpha therapy. Interference with MIF action thus offers potential therapeutic advantages that have not been observed with previous anti-cytokine interventions. The objective of this application is to understand the mechanism by which MIF contributes to the overwhelming inflammatory response that produces shock. Our central hypothesis is that septic shock is the result of genetic predisposition to overproduction of MIF, coupled with M1F activation of cells by binding to the surface protein, CD74. We have formulated this hypothesis on the basis of our studies showing, first, that antiMIF prevents shock in relevant models of bacterial sepsis, second, that MIF is encoded by a functionally polymorphic gene, and third, that MIF initiates signal transduction by
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binding to CD74. The rationale for this proposed research is that once it is known how MIF expression leads to septic shock, then new and selective approaches for therapeutic intervention may be devised. We will test our hypothesis by pursuing the following three specific aims: 1) Define the Frequency of the Low and High Expression M/f Alleles in Patients with Pneumonia, and Determine if the High Expression Alleles are Associated with Septic Shock. Our working hypothesis is that high expression M/f alleles wilt be over-represented in patients with pneumonia who develop shock. 2) Define the Mechanism of MIF Signal Transduction. Our working hypothesis is that CD74 transduces MIF signals by recruiting an additional protein(s) into the signaling complex. 3) Determine the Biological Significance of the MIF-CD74 Interaction. Our working hypothesis is that anti-CD74 mAb and soluble CD74 protein (sCD74) will protect mice from lethal septic shock. The proposed research is innovative because it capitalizes on two recent and potentially unifying findings, a polymorphism in the M/fgene that affects its level of expression, and a cell surface protein (CD74) that mediates MIF signal transduction. These results will be significant because they will provide a better understanding of the molecular pathways responsible for the development of sepsis in susceptible individuals. Website: http://crisp.cit.nih.gov/crisp/Crisp_Query.Generate_Screen •
Project Title: MODELING THE ACUTE INFLAMMATORY RESPONSE Principal Investigator & Institution: Chow, Carson C.; Assistant Professor; Mathematics; University of Pittsburgh at Pittsburgh 350 Thackeray Hall Pittsburgh, Pa 15260 Timing: Fiscal Year 2002; Project Start 01-AUG-2002; Project End 31-JUL-2006 Summary: (provided by applicant): The acute inflammatory response is a cascade of cellular and molecular events that takes place in the body after a traumatic injury or an infection. This response involves the immune, endocrine and neurological systems and aims to eliminate damaging agents and restore the body back to equilibrium. The clinical manifestation of this response is called the systemic inflammatory response syndrome (SIRS) or sepsis in the case of infection. There are approximately threequarters of a million cases of SIRS severe enough to warrant hospitalization in the United States each year. Although much has been learned in the last several years on the molecular and cellular mechanisms of SIRS, this knowledge has not translated into improved outcome prediction or treatments. We hypothesize that a major reason effective treatments have not been developed is that a good understanding of the global dynamical behavior of the acute inflammatory response is lacking. We propose to address this shortcoming by developing biologically accurate mathematical models of the acute inflammatory response. These models will be tested and calibrated with carefully designed animal experiments in an iterative procedure that relies heavily on detailed statistical analysis. More specifically, we propose to 1) develop a hierarchy of mathematical models, each designed to address a specific set of questions; 2) refine and validate the mathematical models through an iterative process of experimentation, statistical analysis, and model development; and 3) analyze the various modes of behavior in the mathematical models and use these modes to make predictions of outcomes in different experimental scenarios. The long-term goal of this study is to provide a rational basis for the design of therapies to combat SIRS as well as to aid in patient management. Website: http://crisp.cit.nih.gov/crisp/Crisp_Query.Generate_Screen
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Project Title: MOLECULAR PROBES OF THE MECHANISM OF CYTOCHROME P450 Principal Investigator & Institution: Groves, John T.; Hugh Stott Taylor Professor of Chemistry; Chemistry; Princeton University 4 New South Building Princeton, Nj 085440036 Timing: Fiscal Year 2002; Project Start 01-DEC-1985; Project End 31-MAR-2003 Summary: The central theme of this program involves studies of the mechanisms of action of cytochrome P450 enzymes and nitric oxide synthase. The principal approach is the synthesis and characterization of reactive iron porphyrin species that are thought to be intermediates in the catalytic cycles and to relate the interconversions of these species toward a molecular understanding of these proteins. Cytochrome P450 is the central protein involved in drug detoxification and hormone metabolism while nitric oxide and peroxynitrite. Synthetic metalloporphyrins can be employed as probes to intervene in these processes in diagnostic ways. Thus, these agents may prove to be significant tools for elaborating the biology of superoxide, peroxynitrite and NO. These same metalloporphyrins have shown impressive activity in animals suggesting their application as pharmaceutical agents. Our effort seeks to provide a foundation of mechanistic and kinetic information which can be applied to in vitro models, cell culture studies and whole animal models of specific disease states such as ischemia-reperfusion, sepsis and autoimmune diseases. Experiments are aimed at determining what reactive intermediates are formed and what their biological targets are likely to be. The elaboration of these will processes will facilitate the design of metal complexes for the catalytic decomposition of peroxynitrite and these other species. The studies of nitric oxide synthase aim to illuminate the range of mechanisms of NOS and to seek out new oxidation processes that may help with the rational development of NOS inhibitors. Peroxoiron(III) porphyrin species are now readily accessible in aqueous solution at ambient temperature. Rapid kinetic techniques have been developed to study the reactivity of these species. A central question is how to modulate the chemistry of these complexes between nucleophilic and electrophilic pathways that are essential to understand the variety of P450 mediated processes. Synthetic and semi-synthetic phospholipid assemblies are used to model and understand the larger scale events in P450 and NOS action and to probe the permeability of membranes to reactive oxygen species. Membrane binding by amphiphilic metalloporphyrins is extraordinarily sensitive to pH. This is an unprecedented observation that may have immediate application to the design of metallopharmaceuticals. Website: http://crisp.cit.nih.gov/crisp/Crisp_Query.Generate_Screen
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Project Title: MUSCLE LACTATE PRODUCTION IN SEPSIS Principal Investigator & Institution: James, J H.; Res Ass Prof; Surgery; University of Cincinnati 2624 Clifton Ave Cincinnati, Oh 45221 Timing: Fiscal Year 2002; Project Start 01-MAY-1997; Project End 31-MAR-2005 Summary: The long-term objective of this work is to understand the changes in metabolism in sepsis in order to improve the therapy of septic patients. Sepsis results in a hypermetabolic state in which many aspects of carbohydrate metabolism are abnormal: enhanced peripheral glucose uptake and utilization, hyperlactacidemia, increased gluconeogenesis, depressed glycogen synthesis, glucose intolerance and insulin resistance. Traditionally, high circulating lactate concentration has been interpreted as tissue hypoxia or mitochondrial dysfunction. However, therapy to improve tissue perfusion does not always prevent lactate accumulation. Current
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understanding of energy metabolism cannot explain persistent glycolysis by welloxygenated tissues. High epinephrine levels in sepsis may cause the characteristic changes in carbohydrate metabolism through stimulation of the Na+-K+ pump in skeletal muscle. The stimulation of glycogen breakdown and lactate production in muscle by epinephrine may be closely tied to stimulation of the Na+, K+-ATPase, implying that muscle energy metabolism takes place in separate glycolytic and oxidative compartments. ATP consumption by the Na+, K+-ATPase appears to be the primary influence on ATP production in the glycolytic compartment. This proposal aims to explore these relationships in greater detail, both in septic and nonsepticrats. Studies will combine in vivo and in vitro assessments of Na+-K+ pump activity, membrane recruitment, glycolysis, glycogenolysis, ATP content and membrane permeability to Na+ and K+. The central role of epinephrine in sepsis-induced metabolic derangements will be examined in two ways (i) chronic infusion of epinephrine using implantable minipumps and (ii) chronic infusion of the beta-adrenergic blockers in sepsis. Studies in vitro will examine the persistence of epinephrine's effects on glycolysis and Na+, K+ATPase activity after beta-blockade has occurred. Results of these studies will clarify metabolic relationships that are important both in health and disease. Website: http://crisp.cit.nih.gov/crisp/Crisp_Query.Generate_Screen •
Project Title: MUSCLE PROTEIN TURNOVER AND AMINO ACID UPTAKE IN SEPSIS Principal Investigator & Institution: Hasselgren, Per-Olof J.; Beth Israel Deaconess Medical Center St 1005 Boston, Ma 02215 Timing: Fiscal Year 2002; Project Start 01-JAN-1987; Project End 31-JUL-2004 Summary: Previous studies suggest that sepsis-induced muscle catabolism reflects ubiquitin-proteasome-dependent degradation of myofibrillar proteins regulated by glucocorticoids. Because intact myofibrils are not degraded by the proteasome, it is possible that actin and myosin are dissociated from the myofibrils before they are ubiquitinated and degraded by the proteasome. We will test the hypotheses: 1) sepsis results in glucocorticoid-mediated calcium/calpain-dependent Z-band disintegration and release of myofilaments in skeletal muscle; 2) sepsis results in increased N-end rule pathway-dependent ubiquitination and breakdown of muscle proteins and upregulated expression and activity of the ubiquitin- conjugating enzyme E2/14k and ubiquitin ligase E3alpha; 3) sepsis- induced muscle cachexia can be inhibited by proteasome blocker in vivo; 4) muscle cachexia in patients with sepsis is associated with increased expression and activity of calpains, release of myofilaments and upregulated protein breakdown in the N-end rule pathway. A septic model in rats consisting of cecal ligation and puncture is used in the majority of experiments. Total and myofibrillar protein breadkdown rates are measured in incubated muscles by determining net release of tyrosine and 3-methylhistidine respectively. Integrity of sarcomeric Z-bands is studied by electron microscopy. Gene and protein expression of calpain and calpastatin are determined by Northern and Western blot analysis, respectively. The role of calcium/calpain-dependent proteolysis is assessed by the effect of dantrolene and diltiazem on sepsis-induced morphologic and metabolic changes. The role of glucocorticoids in sepsis- induced changes in muscle calcium levels and release of myofilaments is determined by the glucocorticoid receptor antagonist RU38486. To test the role of the N-end rule pathway, expression and activity of E2/14k and E3alpha are determined and specific E3albha inhibitors are used in a cell- free system. Similar determinations are performed in muscle from patients with sepsis. The proposal is novel because it suggests that muscle cachexia during sepsis is caused by two distinct
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mechanisms, i.e., calcium/calpain-dependent release of myofilaments from the sarcomere followed by ubiquitination of myofilaments in the N-end rule pathway and subsequent degradation of ubiquitinated filaments by the 26S proteasome. The hypothesis implies two levels at which sepsis-induced muscle cachexia may be prevented/treated, i.e., inhibition of myofilament release by treatment with a calcium antagonist and inhibition of ubiquitin/proteasome-dependent degradation of the released myofilaments by a proteasome blocker. Website: http://crisp.cit.nih.gov/crisp/Crisp_Query.Generate_Screen •
Project Title: NEUTROPHIL ACTIVATION IN ACUTE LUNG INJURY Principal Investigator & Institution: Abraham, Edward; Roger Sherman Mitchell Professor; Medicine; University of Colorado Hlth Sciences Ctr P.O. Box 6508, Grants and Contracts Aurora, Co 800450508 Timing: Fiscal Year 2002; Project Start 10-JUL-2000; Project End 31-MAY-2005 Summary: (adapted from the application): Acute lung injury is frequently associated with sepsis or blood loss. In preliminary studies, The investigators found that hemorrhage or endotoxemia cause rapid increases in the expression of proinflammatory cytokines IL-1, TNF, and MIP-2, and in activation of the transcriptional regulatory factors NF-kB and CREB in lung neutrophils. After endotoxemia, but not hemorrhage, adrenergic blockade increases expression of proinflammatory cytokines by lung neutrophils, adrenergic blockade increased CREB and decreased NF-kB activation after hemorrhage, but decreased CREB and increased NF-kB activation after endotoxemia. After hemorrhage, but not endotoxemia, the increased expression of proinflammatory cytokines by lung neutrophils was inhibited by blockade of xanthine oxidase. After hemorrhage, but not endotoxemia, blockade of xanthine oxidase increases CREB activation, but does not prevent NF-kB activation in lung neutrophils. They hypothesize that distinct mechanisms lead to activation of activation of NF-kB and CREB in lung neutrophils after hemorrhage or endotoxemia, modulating their ability to affect subsequent transcription of proinflammatory cytokines involved in the development of acute lung injury. The specific aims are to determine: 1) the mechanisms involving alterations in nuclear and cytoplasmic I-kBs, NF-kB relevant kinases, and NF-kB subunit composition, which lead to hemorrhage or endotoxemia-induced activation of NF-kB in lung neutrophils; 2) the mechanisms involving CREB relevant kinases, CREB phosphorylation, CREM and ICER expression, which lead to hemorrhage or endotoxemia-induced activation of CREB in lung neutrophils; 3) the effects of hemorrhage or endotoxemia on binding interactions between CREB, NF-kB, and CBP in lung neutrophils; and 4) the mechanisms by which reactive oxygen intermediates or catecholamines modulate NF-kB and CREB activations as well as their interactions with CBP in lung neutrophils after hemorrhage or endotoxemia, and the effects of such modulation of NF-kB and CREB activity on the development of acute lung injury. Website: http://crisp.cit.nih.gov/crisp/Crisp_Query.Generate_Screen
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Project Title: NEUTROPHILS AND STAPHYLOCOCCUS AUREUS Principal Investigator & Institution: Gresham, Hattie D.; Associate Professor; Molecular Genetics & Microbiol; University of New Mexico Albuquerque Controller's Office Albuquerque, Nm 87131 Timing: Fiscal Year 2002; Project Start 01-SEP-2000; Project End 31-MAY-2005 Summary: Description (Adapted from applicant's abstract): Staphylococcus aureus is a major human pathogen causing significant morbidity and mortality in both community-
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and hospital-acquired infections. Concern over the emergence of multidrug resistant strains, particularly strains which lack sensitivity to all currently available antibiotics, has renewed interest in understanding the virulence mechanisms of this pathogen at the molecular level and in elucidating host defense elements which either provide protection from or which limit infection. Neutrophils (PMN) have long been thought to provide significant host defense against S. aureus infection. However, our studies of S. aureus-induced peritonitis and sepsis in mice have suggested that PMN have both a protective and a deleterious role. In order to demonstrate that PMN contribute to the pathogenesis of S. aureus infection, we have used multiple approaches which either limit or promote PMN migration into the infectious site. Our data indicate that excessive numbers of PMN and elevated levels of a C-X-C chemokine, MIP-2, at the site of a S. aureus infection create an environment which leads to enhanced extracellular replication of the pathogen and its intracellular survival in PMN to the detriment of the host; that PMN isolated from this environment are sufficient to establish infection in naive animals; that some of the bacteria inside these infected PMN are in endosomes with partially or fully degraded membranes; and that two regulatory loci mutants (agrand sar-) which lack the expression of several virulence factors are less able to survive and/or avoid clearance in the presence of excess PMN and MIP-2. We hypothesize that S. aureus manifests as a virulence determinant the ability to exploit the host's inflammatory response in order to enhance its survival. Moreover, we hypothesize that exogenous modulation of the inflammatory response is sufficient to alter the susceptibility of the host to infection. To test this hypothesis, we will pursue the following specific aims: #1) determine the number of PMN necessary for protection and for their deleterious role in two models of S. aureus infection; #2) define the contribution of C-X-C chemokines, the CXCR2 receptor, and specific virulence factors expressed by S. aureus to the creation of the environment which leads to both enhanced extracellular replication and intracellular survival of the pathogen; #3) elucidate known virulence factors whose genes are activated both in vivo and in vitro specifically in the presence of C-X-C chemokines and PMN; and #4) determine the mechanism of uptake and the intracellular locale of wild-type and isogenic mutants of S. aureus taken up both in vivo and in vitro by C-X-C chemokine-stimulated PMN. Website: http://crisp.cit.nih.gov/crisp/Crisp_Query.Generate_Screen •
Project Title: NEW POINT OF CARE SYSTEM FOR BURN WOUND TREATMENT Principal Investigator & Institution: Denvir, Adrian J.; Lynntech, Inc. College Station, Tx 77840 Timing: Fiscal Year 2003; Project Start 03-JUN-2003; Project End 31-MAY-2004 Summary: (provided by applicant): Each year in the United States, two million people suffer burn injuries resulting in approximately 100,000 - 300,000 hospitalizations. Of these two million, more than 12,000 will eventually die of burn-related infections. People suffering from burn wounds have lost a portion of their integument, diminishing one of the body's main defenses against bacterial infection. In addition, the necrotic tissue in the burn eschar, combined with the presence of serum proteins, provides a rich culture medium for microorganisms. Added to the loss of integument is the adverse affect of thermal injury on both local and systemic immunity. For these reasons, infections and sepsis are the most common cause of death among hospitalized burn patient. New methods of burn wound management require a cocktail of antimicrobial and antibiotic agents to fight infection however; they are becoming less effective due to resistant strains of microorganisms. In this Phase I effort we will demonstrate the feasibility of a fundamentally new method for burn wound care. The point of care (POC) system will
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be designed to generate sterile saline or water to moisten or wet the dressings for both the eschar as well as the graft after excision and deliver a broad based antimicrobial agent effective, against gram positive and gram-negative bacteria and fungi to the wound surface. Website: http://crisp.cit.nih.gov/crisp/Crisp_Query.Generate_Screen •
Project Title: P20, MOLECULAR SHORTSTOP FOR INFLAMMATORY LUNG DISEASES Principal Investigator & Institution: Brigham, Kenneth L.; Professor of Medicine; Generx+, Inc. 3200 West End Ave, Ste 500 Nashville, Tn 372011322 Timing: Fiscal Year 2002; Project Start 20-MAY-2000; Project End 31-JUL-2004 Summary: (Applicant's abstract): The transcription factor CAAT enhancer binding protein beta (C/EBPbeta) is a key factor orchestrating the inflammatory response. Specifically, expression of the genes encoding the pro-inflammatory cytokines IL-6 and on IL-8 are regulated by C/EBPbeta. C/EBPbeta can be either an activator or an inhibitor of inflammation depending on the dominant isoforms produced. Our data indicate that airway epithelial cells in culture terminate production of IL-6 and IL-8 increasing production of inhibitory C/EBPbeta isoform (p20) and that cells that have an exaggerated inflammatory response produce decreased amounts of this isoform. In addition, when animals are given endotoxin there is a decrease in lung production of the inhibitory C/EBPbeta isoform and an increase in production of the activator isoform. Since IL-6 production correlates with severity of sepsis in humans and interventions that decrease IL-6 improve outcome in that setting, we believe that increasing the inhibitory C/EBPbeta isoform in the lungs could be therapeutic for the pulmonary complications of sepsis. Website: http://crisp.cit.nih.gov/crisp/Crisp_Query.Generate_Screen
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Project Title: PATHOGENESIS OF ENDOTOXEMIA-INDUCED ACUTE RENAL FAILURE Principal Investigator & Institution: Schrier, Robert W.; Professor; Medicine; University of Colorado Hlth Sciences Ctr P.O. Box 6508, Grants and Contracts Aurora, Co 800450508 Timing: Fiscal Year 2002; Project Start 01-JAN-1998; Project End 31-MAY-2006 Summary: (provided by applicant): Sepsis and septic shock in-hospital are associated with a high incidence of acute renal failure (ARF) estimated at 20 and 50%, respectively. Moreover, the combination of sepsis and ARF leads to a very high mortality ranging from 50-80%. Understanding the early vasoactive and the later proinflammatory events which cause ARF during endotoxemia would be a major medical advance, thereby allowing the development of pathogenetic-based interventions. This proposal focuses on early vasoactive endotoxemia-related events which cause systemic arterial vasodilation (e.g. nitric oxide, prostaglandins) and the compensatory vasoconstrictors (e.g. norepinephrine, angiotensin, endothelin, thromboxane) which support blood pressure but lead to renal vasoconstriction. This renal vasoconstriction renders the kidney more susceptible to the proinflammatory events of endotoxemia, such as generation of superoxide, peroxynitrite, and cytokines such as interleukin-18. In addition to selective renal denervation, there are now specific inhibitors to test the involvement of inducible nitric oxide synthase, prostaglandin and thromboxane in sepsis. Molecular biological techniques using knockout and transgemc mice will also be used to examine the role of various systemic and renal vasodilators and vasoconstrictors during endotoxemia.
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Understanding the role and interaction between these factors should allow the development of potential interventions in sepsis which could dramatically decrease the incidence of ARF, morbidity and mortality. The availability of potent scavengers of superoxide and antiserum to interleukin-18 also will allow not only the study of early vasoactive events but also the later pro-inflammatory events. Thus, the potential impact of the research on sepsis and sepsis-mediated ARF is substantial. The goal is to unravel the multifaceted events which occur during sepsis, including their interactions, so that effective interventions can be developed and tested in humans with sepsis. The ultimate goal therefore is to understand and prevent the ARF, morbidity and mortality associated with sepsis. Website: http://crisp.cit.nih.gov/crisp/Crisp_Query.Generate_Screen •
Project Title: PATHOGENIC MECHANISMS OF ANAEROBES IN SEPSIS Principal Investigator & Institution: Kasper, Dennis L.; Associate Director; Brigham and Women's Hospital 75 Francis Street Boston, Ma 02115 Timing: Fiscal Year 2003; Project Start 01-MAY-1997; Project End 31-DEC-2007 Summary: (provided by applicant): Abscess formation is a classic host response to bacteria during sepsis. Certain bacterial species, particularly Bacteroides fragilis and Staphylococcus aureus are predisposed to induce abscesses. The essential bacterial virulence factor required for abscess induction by these two pathogens is a capsular polysaccharide with a zwitterionic charge motif. Zwitterionic polysaccharides (ZPS) induce the host to form abscesses by their ability to activate T cells initiating a proinflammatory Th1 cytokine response. In contrast to the immunologic paradigm defining polysaccharides as T cell independent antigens, ZPS activate T cells in vitro as well as in vivo when incubated with antigen presenting cells (APC). There is currently no immunologic model that describes how purified polysaccharides can activate T cells. ZPS are internalized and can be detected in lysates of intracellular vesicles from the APC. Blocking of endosomal acidification results in the failure of ZPS to activate T cells. ZPS recovered from endosomal vesicles has a substantially reduced molecular size, indicating processing. We have demonstrated that MHC class II DR appears to be the molecule used by the APC to present ZPS to the T cell and that TCR alpha beta( is required for T cell activation. We hypothesize that ZPS are internalized and cycle through the APC, and that this process is required for presentation of the ZPS to the T cell. We intend to define a novel immunologic paradigm that describes how an important class of biologic molecules (carbohydrates) is recognized by the cell-mediated immune system. This will be done by investigating the cellular pathway by which ZPS cycle through the APC and activate CD4+ T cells. We have defined four specific aims: 1) Determine how ZPS are altered within the endocytic pathway and define the molecular requirements for ZPS-mediated T-cell activation; 2) Investigate the vesicular trafficking and intracellular interactions of ZPSs in the endocytic pathway; 3) Characterize the binding interactions of the MHC class II DR molecule with ZPS; 4) Determine whether T-cell activation results from "processed antigen" presentation or superantigen presentation and whether the processing of ZPS uses the same pathway as protein processing. The delineation of a mechanism for carbohydrate processing and presentation has broad relevance to the fields of microbiology and immunology and could lead to new concepts for enhancing T cell recognition of other polysaccharides. Website: http://crisp.cit.nih.gov/crisp/Crisp_Query.Generate_Screen
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Project Title: PH CONTROL OF PROTEIN SYNTHESIS Principal Investigator & Institution: Garlick, Peter J.; Professor and Director of Research; Surgery; State University New York Stony Brook Stony Brook, Ny 11794 Timing: Fiscal Year 2002; Project Start 01-JUN-2000; Project End 31-MAY-2004 Summary: Patients with renal failure, diabetic ketosis, severe trauma or sepsis often become acidotic. Moreover, they are also susceptible to body protein wasting. Experimental acidosis has been shown to result in negative nitrogen balance, and body protein loss in acidotic patients can be alleviated by normalization of pH. Previous work has concentrated on the effects of pH on protein degradation and no systematic studies have been performed on pH and protein synthesis. However, measurements in animals and human volunteers have shown that both metabolic and respiratory acidosis are associated with depressed rates of synthesis of skeletal muscle protein and serum albumin. The purpose of the present proposal is therefore to characterize the relationship between protein synthesis rates in tissues, especially muscle and liver, of rats and humans in relation to changes in intra- and extracellular pH, to confirm the hypotheses: (i) that changes in blood pH, both by metabolic and respiratory means, modify rates of tissue protein synthesis and gene expression and contribute to the protein wasting of patients with acidosis, and (ii) that these effects of pH operate directly via the intracellular pH and do not involve extracellular mediators such as hormones. The studies will employ measurements of rates of protein synthesis in tissues of rats and human volunteers to characterize the responses to changes in intra- and extracellular pH induced by metabolic and respiratory means and to confirm that these effects are rapid, operate over the full range of pH from acidosis to alkalosis and are independent of oxygen supply. In the animals, the responses of gene expression in the liver will be investigated by determining the levels of mRNA for 3 liver proteins. In particular, the treatments studied will produce different changes in the infra- and extracellular pH. Measurements of intracellular pH will be made by nuclear magnetic resonance, to confirm the hypothesis that changes in protein synthesis are determined by the intracellular rather than extracellular pH. A separate set of studies will be performed in isolated tissues and cells, to determine whether responses to changes in pH are direct or act through circulating hormones. The cellular and molecular mechanisms through peptide chain initiation and initiation factors eIF-2 and eIF-4 will also be identified. Finally, the effect of changes in pH on tissue protein synthesis will be measured in human volunteers and hemodialysis patients, to identify the role of pH control of protein synthesis in health and in protein wasting conditions with abnormalities of pH control. Website: http://crisp.cit.nih.gov/crisp/Crisp_Query.Generate_Screen
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Project Title: PHAGOCYTE RECEPTORS FOR LIPID A Principal Investigator & Institution: Golenbock, Douglas T.; Chief, Division of Infectious Diseases &; Medicine; Univ of Massachusetts Med Sch Worcester Office of Research Funding Worcester, Ma 01655 Timing: Fiscal Year 2002; Project Start 01-JAN-1999; Project End 31-MAR-2004 Summary: Gram-negative bacterial sepsis is a medical catastrophe, claiming the lives of 35-40 percent of affected patients. There has been little progress in improving the poor outcome of sepsis, due to our inadequate understanding of the basic pathophysiology of the syndrome. Sepsis results from a cascade of life-threatening host responses beginning with the interaction of bacterial lipopolysaccharide (LPS) and the LPSreceptor complex on effector cells. CD14 appears to be the LPS-binding component of
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this complex. The exact components of the complex, and the mechanism of signal transduction, are uncertain. Two Toll-like receptors (TLRs), TLR2 and TLR4, function as LPS signal transducers. Expression of TLR4, but not TLR2, is obligatory for sensitive responses to LPS. TLR2 functions as a pattern recognition molecule for bacterial products (including LPS) from multiple types of bacteria. LPS-receptor specificity resides in TLR4: expression of TLR4 defines a highly specific pharmacology to lipid A and its analogs. The central hypothesis of this proposal is that LPS is recognized by a multimeric receptor consisting of CD14 and of a complex of TLRs and accessory molecules. TLR4 is the predominant LPS receptor in most cells; TLR2 may be important for sCD14/LPS recognition. Upon ligand binding, TLRs multimerize and engage signaling molecules similar or identical to those used by the IL-1 receptor. LPS antagonists may function by preventing TLR multimerization. We propose three Aims to test this hypothesis. 1. To analyze available known TLRs (including TLRs 1, 3, 5, 6 and 7) and chimeric TLR constructs as potential LPS receptor components. To identify regions of TLR4 that are necessary for specific ligand recognition. To determine if epitope-tagged TLRs multimerize, and if multimerization is inhibited by LPS antagonists. 2. To use [32P]-lipid IVa to analyze lipid A binding to TLRs, especially TLR4. 3. To identify new genes in the LPS signal transduction pathway using somatic cell mutagenesis and complementation analysis approaches. The discovery of Toll-like receptors as mediators of signal transduction for both Gram-positive and Gram-negative bacteria suggests that there are common signaling elements that can be discovered and targeted for the rational design of novel therapeutic agents for sepsis, an thus reduce the appalling morbidity and mortality associated with this disease. Website: http://crisp.cit.nih.gov/crisp/Crisp_Query.Generate_Screen •
Project Title: PHYSIOLOGIC REGULATION OF HEMATOPOIESIS BY NOTCH Principal Investigator & Institution: Carlesso, Nadia; Assistant Professor of Medicine; Massachusetts General Hospital 55 Fruit St Boston, Ma 02114 Timing: Fiscal Year 2002; Project Start 15-AUG-2001; Project End 31-JUL-2005 Summary: (provided by applicant): Little is known about the mechanisms that regulate, at the cellular level, the process of hematopoietic cell self-renewal and differentiation, in particular during physiologic responses to stress, such as inflammation and infection. The overall objective of this project is to study the physiologic role of Notch receptors in maintaining bone marrow homeostasis, in particular during inflammatory stress. The main hypotheses to be tested in our study are: I) whether Notch activation favors the decision of self-renewal over differentiation at the stem and progenitor cell level, and 2) whether this represents an important regulatory function during bone marrow physiologic response to inflammation and infection. We hypothesize that Notch activation may balance the prodifferentiative effects of inflammatory cytokines, permitting expansion of the proliferating pool of progenitor cells required to respond to physiologic stress and preventing the stem cell pool from exhaustion. Specific Aims: to test these hypotheses we plan to: (1) determine the impact of Notch gain-of function and (2) Notch loss-of function, on self-renewal and expansion potential of stem and progenitor cells; (3) evaluate the level of Notch pathway activation in BM cells during inflammatory stress; (4) determine the consequences of Notchi loss-of function during bone marrow response to inflammatory stress. Research Design and Methods: We will use different and complementary in vivo models: a) xenotransplantation experiments of human cells, engineered to express constitutive forms of Notch 1 or the ligand J2, will be carried out into NOD/S CID mice; b) transgenic mice engineered to express the Notch antisense will be used to evaluate the effects of Notch loss-of function; c) the "mouse full
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skin thickness burn model" (sepsis model) will be used as a model of inflammation to evaluate BM response and Notch function. Significance: These studies are intended to yield insight into the physiologic mechanisms that regulate adult hematopoiesis. We believe that a better comprehension of these events is crucial for the understanding of the biology of hematopoietic disorders and essential for the development of novel therapeutic strategies, in particular those targeted to ex-vivo expansion of human hematopoietic stem cells. Website: http://crisp.cit.nih.gov/crisp/Crisp_Query.Generate_Screen •
Project Title: PILOT--MATERNAL ALCOHOL ABUSE AND ITS EFFECTS ON PREMATURE INFANTS Principal Investigator & Institution: Gauthier, Theresa W.; Emory University 1784 North Decatur Road Atlanta, Ga 30322 Timing: Fiscal Year 2003; Project Start 01-FEB-2003; Project End 31-DEC-2007 Summary: Despite the well known neurological effects of alcohol on the developing fetus, its use during pregnancy remains a significant problem in our country. Even with modern neonatal intensive care units, chronic lung disease and bacterial sepsis continue to cause significant morbidity and mortality for the very low birth weight premature newborn. We have expanded the traditional focus of fetal alcohol exposure on the central nervous system to two other developing organ systems of the fetus, namely the lung and immune system. Recent prospective data implicates a history of alcohol abuse as the first reported co-morbid variable significantly increasing the incidence and severity of acute respiratory distress syndrome in adults. Additionally, alcohol exposure is associated with a decrease in lung antioxidant status, particularly glutathione. Glutathione is an essential antioxidant in the epithelial lining fluid of the lung. A reduction in alveolar glutathione, as seen in the premature infant, leaves the lung susceptible to increased pulmonary oxidative injury. We have novel experimental data describing reduced pulmonary glutathione, impairment of alveolar type II epithelial cell function and impaired cell function and impaired surfactant homeostasis in fetal guinea pig lungs exposed to alcohol in utero. Additionally, in a logistic regression analysis of 872 term newborns, we have demonstrated that maternal excessive alcohol use of 7 drinks/wk in either the 3 months prior to conception of the 2nd trimester significantly increased the risk of newborn infection by approximately 3 fold. Because of these data implicating adverse effects of fetal alcohol exposure on the developing fetus, we hypothesize the following: (1) alcohol exposure in (CLD), and (2) fetal alcohol exposure impairs fetal immune function thereby increasing the risk of infection, particularly late onset sepsis, in the premature newborn. We will address these hypotheses in the pilot clinical study by 1: Screening all women who deliver premature infants weighing <1500 gms for alcohol use during pregnancy with an extensive questionnaire. 2: Perform outcome analysis of these infants comparing alcohol-exposed to non-exposed premature infants, investigating the primary outcomes of chronic lung disease and late onset sepsis. Website: http://crisp.cit.nih.gov/crisp/Crisp_Query.Generate_Screen
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Project Title: PREVENTION OF APOPTOTIC CELL DEATH IN SEPSIS BY BCL-2 Principal Investigator & Institution: Hotchkiss, Richard S.; Professor; Anesthesiology; Washington University Lindell and Skinker Blvd St. Louis, Mo 63130 Timing: Fiscal Year 2002; Project Start 01-AUG-1999; Project End 31-JUL-2003
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Summary: (adapted from the abstract): Sepsis is defined as the systemic inflammatory response from infection and is a major cause of morbidity and mortality with an annual death rate of over 175,000 people in the United States alone. The host response to sepsis is an intricate interplay of numerous inflammatory and anti-inflammatory processes including the coagulation cascade, complement system, kinin system, and many others. Immunologic defenses are activated in sepsis as well; and there is recruitment and activation of granulocytes, lymphocytes, and monocytes. Recent studies demonstrate that sepsis causes a marked decrease in circulating lymphocytes which is accompanied by extensive apoptosis of lymphocytes in tissues throughout the body. Although it is speculated that sepsis-induced apoptosis of lymphocytes may be beneficial in sepsis by down-regulating the inflammatory response, it is also possible that loss of lymphocytes is excessive and impairs the ability of the host to eradicate the infection. Recently, the anti- apoptotic protein BCL-2 has been shown to prevent cell death from a remarkable number of diverse stimuli including hypoxia, ionizing radiation, oxidant injury and excitotoxins. The aims of this investigation are to pursue their initial observations demonstrating that transgenic mice which selectively overexpress BCL-2 in T lymphocytes have complete protection against lymphocyte apoptosis and improved survival in sepsis. These findings indicate that strategies which prevent lymphocyte death in sepsis may improve survival. Transgenic mouse constructs in which BCL-2 is selectively overexpressed in T, B, or both T and B lymphocytes will be employed in sepsis. Lymphocyte transfer experiments in Rag-1 mice, which are totally deficient in mature T and B cells, will be examined also. Rag-1 mice will be transfused with T or B lymphocytes that overexpress BCL-2 and effects on apoptosis and survival recorded. Mechanisms of BCL-2's protective effect will be investigated as well. Finally, autopsy sample from patient who died of sepsis or other causes will be examined to correlate BCL-2's expression with lymphocyte apoptosis and gain insight into possible mechanisms of apoptosis such as caspase activation and cytochrome c release. Website: http://crisp.cit.nih.gov/crisp/Crisp_Query.Generate_Screen •
Project Title: PROTEASE ACTIVATED RECEPTOR-1 SIGNALING BY PROTEIN C Principal Investigator & Institution: Riewald, Matthias; Scripps Research Institute Tpc7 La Jolla, Ca 92037 Timing: Fiscal Year 2003; Project Start 01-AUG-2003; Project End 31-JUL-2007 Summary: (provided by applicant): Protein C (PC) is activated on the surface of endothelial cells by the thrombin-thrombomodulin complex and the generated activated PC (APC) inhibits blood coagulation by down regulating prothrombin activation in a negative feedback loop. In addition to its role as an anticoagulant, APC has major antiinflammatory effects that led to its recent approval for treatment of patients with severe sepsis. The molecular basis for APC's important protective effects is incompletely understood. This application is based upon our recent identification of proteaseactivated receptor 1 (PAR1) and endothelial cell PC receptor (EPCR) as part of a novel APC signaling pathway in endothelial cells. The general hypothesis is that PARldependent signaling of the PC system in endothelial cells limits inflammatory responses and reduces lethality in sepsis. PAR1 can be efficiently activated by thrombin and Aim#1 is to define how autocrine PAR1 signaling by APC is linked to PC activation on the cell surface. Experiments will analyze whether activation of EPCR-bound PC by thrombin-thrombomodulin leads to enhanced APC signaling and what the role of membrane compartmentalization of PC pathway components and PAR1 is. Aim#2 is to define the role of PAR1, EPCR, and thrombomodulin in APC-dependent effects on gene expression, including the down regulation of genes involved in inflammatory and
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apoptotic pathways in activated endothelial cells. Micro array analysis will identify downstream targets of protective PC pathway signaling and adenovirus-mediated expression of mutant EPCR and thrombomodulin will define the roles of these cofactors. Aim#3 is to demonstrate an in vivo role of PAR1 activation by the PC pathway. Transgenic mice will be generated with endothelial cell-specific expression of either wild type PAR1 or a variant PAR1 that can be activated by APC but not thrombin. Effects of APC administration on inflammatory responses and lethality in a model of septicemia will be analyzed in the mice and the results are expected to define the role of endothelial PAR1 in systemic inflammation and in mediating protective effects of APC. Website: http://crisp.cit.nih.gov/crisp/Crisp_Query.Generate_Screen •
Project Title: PROTEASE-ACTIVATED RECEPTORS IN INFLAMMATION Principal Investigator & Institution: Coughlin, Shaun R.; Professor; Cardiovascular Research Institute; University of California San Francisco 500 Parnassus Ave San Francisco, Ca 941222747 Timing: Fiscal Year 2003; Project Start 01-JAN-1998; Project End 31-MAR-2007 Summary: (provided by applicant): The long term goal of this proposal is to define the role of protease-activated receptors (PARs) in inflammatory responses. PARs mediate cellular responses triggered by coagulation proteases and other proteases that are generated or released at sites of tissue injury. The effects of PAR activation in different cell types, largely defined in culture, suggest that PARs may help orchestrate a coordinated response to tissue injury that includes hemostasis, inflammation, and perhaps even regulation of the adaptive immune response. Local PAR activation by proteases generated at sites of local bacterial inoculation may protect against spread of bacteria by promoting both recruitment of leukocytes and microvascular thrombosis. However, more regional or systemic activation of PARs as may occur in the setting of tissue ischemia or sepsis may promote tissue damage by the same mechanisms. Toward testing these hypotheses, we have generated knockout mice for the known PARs as well as relevant double knockouts. Par4 -/- mice, for example, have no platelet responses to thrombin. Mice lacking both PAR1 and PAR4 may have lost thrombin signaling in all cell types. Using such mice and cells derived from them, we shall determine which PARs are responsible for mediating responses to coagulation proteases and other proteases in endothelial and other cell types, the extent to which different PARs in the same cell serve redundant functions, and the potential roles for endothelial PAR activation in vivo. We shall then go on to determine the effects of loss of PAR function in models of a) local bacterial infection and dissemination of same, b) systemic inflammatory response syndromes induced by endotoxin and by pancreatitis, and c) ischemia/infarction. Endpoints will include survival; markers of platelet and fibrin deposition and leukocyte accumulation in tissues; vascular permeability and edema; cytokine production; organ histology; and in a hind-limb ischemia-reperfusion model, reflow and infarct size. Lastly, because PARs sense tissue injury and hence "immunological danger," we shall ask whether PARs influence the decision to mount an adaptive immune response. These studies may point to new strategies for modulating injurious inflammatory responses in man. Website: http://crisp.cit.nih.gov/crisp/Crisp_Query.Generate_Screen
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Project Title: PROTECTIVE EFFECTS OF ANTI-C5A IN SEPSIS Principal Investigator & Institution: Ward, Peter A.; Godfrey D. Stobbe Professor and Chairman; Pathology; University of Michigan at Ann Arbor 3003 South State, Room 1040 Ann Arbor, Mi 481091274
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Timing: Fiscal Year 2002; Project Start 15-AUG-2002; Project End 31-MAY-2006 Summary: (provided by applicant): On the basis of our studies to date using the experimental model of sepsis induced by cecal ligation/puncture (CLP) in rats, serious impairment of innate immunity develops. This results in what appears to be a C5adependent defect in assembly of NADPH oxidase and defective phagocytic function of neutrophils. These defects can be reproduced by in vitro exposure of neutrophils to concentrations of C5a found in sepsis. In the first aim, we will evaluate how in vitro exposure of neutrophils to C5a results in detective signaling pathways: phorbol 12myristate 13-acetate (PMA)-induced activation of phosphokinase C (PKC) which results in assembly of NADPH oxidase; and cell activation by engagement of FcyRs resulting in phagocytic responses. In the second aim, we will evaluate the same signaling pathways in blood neutrophils from CLP animals and determine if treatment with anti-C5a prevents defective signaling. In the third aim, we will determine if treatment of normal rats and mice and CLP rats and mice with anti-C5a compromises innate immunity, as assessed by bacterial clearance (Pseudomonas sp. and Klebsiella sp.) from lungs and evaluate the effects on survival. In the fourth aim, we will employ microarray analysis in CLP rats to define, as a function of time, alterations in global gene expression in organs that are predisposed to injury during sepsis (liver, lungs, kidneys, thymus) and determine if treatment with anti-C5a prevents this pattern of gene expression. It is possible that microarrary analysis will be predictive of organ susceptibility to damage during sepsis. Collectively, these studies should provide important evidence related to the mechanisms by which complement activation during sepsis impairs innate immunity. Website: http://crisp.cit.nih.gov/crisp/Crisp_Query.Generate_Screen •
Project Title: PSEUDOMONAS' EFFECTS ON THE GUT BARRIER FROM SURGERY Principal Investigator & Institution: Alverdy, John C.; Professor of Surgery; Surgery; University of Chicago 5801 S Ellis Ave Chicago, Il 60637 Timing: Fiscal Year 2002; Project Start 01-FEB-2001; Project End 31-JAN-2005 Summary: The mere presence of Pseudomonas aeruginosa in the intestine of critically ill surgical patients is associated with a 70% mortality rate--a 3 fold increase above matched patients who culture negative for this pathogen. We propose that within the intestinal tract of a surgically stressed host, physical and chemical environmental signals cause critical shifts in the virulence phenotype of P. aeruginosa. These effects result in a change in the behavior of intestinal P. aeruginosa, causing this bacteria, upon proper cue, to shift from an indolent colonizer to a life-threatening pathogen. In this proposal we provide strong evidence that a virulence determinant in P. aeruginosa, the PA-I lectin/adhesin, plays a key role in lethal gut- derived sepsis in a surgically stressed host. The hypotheses to be tested in this project are: 1) the PA-I lectin of P. aeruginosa is expressed in vivo in response to environmental cues in the intestinal tract including pH, redox state, and norepinephrine following surgical stress (hepatectomy) 2) the PA-I lectin of P.aeruginosa induces an epithelial permeability defect at the level of the intercellular tight junction resulting in paracellular transport of its lethal cytotoxins, and 3) the PA-I lectin of P. aeruginosa alters epithelial tight junctional permeability by activation of regulatory molecules involved in the expression of occludin, the rate limiting seal of the paracellular pathway. We will test these hypotheses using a novel mouse model of endogenous P. aeruginosa sepsis and cultured intestinal epithelial cells that we have extensively studied. Our specific aims to test these hypotheses are: 1) Determine the expression, location, and function of PA-I in P. aeruginosa harvested
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from different tissue sites in mice following surgical stress (hepatectomy) and cecal injection of live P. aeruginosa and following in vitro manipulation of the its physical microenvironment (pH, redox, osmolality, norepinephrine). 2) Determine the route of transport of the P. aeruginosa cytotoxins, exotoxin A and elastase, across cultured intestinal epithelial cells (Caco-2) in response to purified PA-I and selected mutants of live P. aeurginosa. 3) Explore potential cellular mechanisms of PA-I-induced decreases in intestinal epithelial barrier function. We propose that we should rethink our understanding of the gut theory of sepsis to include mechanisms by which pathogenic bacteria alter their virulence strategies in response to stressful changes in their local environment. Understanding the virulence determinants and cellular mechanisms that pathogens use to adhere to and modify the intestinal epithelial barrier may lead to therapies which can avoid nosocomial infection at a more proximate point in the care of the critically ill. Website: http://crisp.cit.nih.gov/crisp/Crisp_Query.Generate_Screen •
Project Title: QUORUM PATHOGENESIS
SENSING
AND
STAPHYLOCOCCUS
AUREUS
Principal Investigator & Institution: Balaban, Naomi; Assistant Professor; Biomedical Sciences; Tufts University Boston Boston, Ma 02111 Timing: Fiscal Year 2004; Project Start 01-APR-2004; Project End 31-MAR-2006 Summary: (provided by applicant): This proposal is responsive to PA-03-080 on "Biodefense and emerging infectious disease research opportunities". Our aim is to better understand how virulence is regulated in Staphylococcus aureus and to further develop immunotherapy to staphylococcal infections. S. aureus cause food poisoning, sepsis, device (Biofilm) related infections, and multiple diseases like endocarditis and pneumonia. The numerous toxins produced by the bacteria are at the heart of these diseases and cause more than 50,000 deaths each year in the US alone. As food and device-associated infectious organism that can survive in multiple environments, S. aureus is a potential target for bioterrorism. Many of the infective strains are resistant to conventional antibiotics and alternative methods to prevent and treat such infections are mandatory. We discovered that the production of toxins is regulated by quorum sensing mechanisms, where proteins such as RNAIIl activating protein (RAP) are secreted by the bacteria and induce virulence. RAP (native or recombinant) activates S. aureus pathogenesis through the histidine-phosphorylation of TRAP in a still unknown manner. TRAP has been shown to be membrane-associated, but it lacks a predicted transmembrane domain and a kinase domain typical of two component system sensors. Our first hypothesis is that TRAP is bound to the membrane through another protein and that this protein acts as the actual receptor to RAP. Studies proposed in Specific Aim 1 are focused on identifying this receptor. RAP has been shown to serve as an effective vaccine to prevent S. aureus infections. Our second hypothesis is that monoclonal anti RAP antibodies may be used for immunotherapy. This will be tested under Specific Aim 2, using a panel of clones already available. Such immune intervention would greatly benefit people suffering from acute drug resistant infections or surgery-related infections often associated with implantable medical devices. Website: http://crisp.cit.nih.gov/crisp/Crisp_Query.Generate_Screen •
Project Title: REGULATION OF ALS AND ITS ROLE IN THE IGF SYSTEM Principal Investigator & Institution: Boisclair, Yves R.; Animal Science; Cornell University Ithaca Office of Sponsored Programs Ithaca, Ny 14853
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Timing: Fiscal Year 2003; Project Start 26-SEP-1997; Project End 31-MAR-2007 Summary: (provided by applicant): Gene deletion studies have demonstrated the importance of IGF-I and II (IGFs), particularly during fetal life when local IGFs production predominates. After birth, the liver becomes the most important site of IGFs synthesis, resulting in the development of a substantial plasma reservoir. This reservoir is dependent on the postnatal production of the acid labile subunit (ALS), a protein that recruits IGFs and IGF Binding Protein-3 in long-lived ternary complexes. The significance of this reservoir has been uncertain until we showed that ALS and the plasma IGF-I reservoir are required for early postnatal growth and bone development. We now will extend these studies to normal and diseased states of later postnatal life. This is relevant to malnutrition and catabolic illnesses in which decreased plasma IGF-I is associated with erosion of lean mass. Despite this association, IGF-I-based therapies have had limited success, reflecting the need for their incorporation into ternary complexes for effectiveness. Three specific aims wilt be pursued to address the role of ALS and the circulating IGFs reservoir during diseased states. AIM A: IGF-I is a potent positive regulator of skeletal muscle mass. Null ALS mice will be subjected to challenges known to induce changes in plasma IGF-I and to alter the mass of skeletal muscles (i.e., sudden increase in GH, nutritional deficiency or sepsis). AIM B: Humans have 3 times as much plasma IGF-II than IGF-I. In contrast, mice have little IGF-II and null ALS mice have normal carbohydrate homeostasis. To determine the role of ALS in containing the metabolic effects of IGF-II, we will study null ALS mice over-expressing human IGF-II. AIM C: GH stimulates ALS synthesis by increasing transcription. In vitro, this effect is conveyed by STAT5, but the importance of this mechanism remains to be established in vivo. Using null STAT5 mice and liver cells, we will evaluate the contribution of direct and indirect mechanisms mediating the effects of GH on ALS synthesis. Studying the GH-regulation of ALS transcription will provide clues to mechanisms responsible for development of hepatic GH resistance during catabolic diseases. Overall, these studies will significantly advance our understanding of the roles played by ALS and the circulating IGF reservoir in diseases of postnatal life. Website: http://crisp.cit.nih.gov/crisp/Crisp_Query.Generate_Screen •
Project Title: MONOCYTES
REGULATION
OF
INDUCIBLE
GENE
EXPRESSION
IN
Principal Investigator & Institution: Mackman, Nigel; Associate Professor; Scripps Research Institute Tpc7 La Jolla, Ca 92037 Timing: Fiscal Year 2002; Project Start 01-JUL-1992; Project End 31-MAR-2006 Summary: Gram-negative sepsis is a major cause of death in intensive care units in the United States. Sepsis is induced by the presence of pathogenic bacteria in the blood. Monocytes of the host innate immune system orchestrate a rapid response to bacterial lipopolysaccharide (LPS [endotoxin]) by expressing various cytokines and by expressing the procoagulant protein tissue factor (TF), which initiates disseminated intravascular coagulation. Recent studies indicate that administration of anticoagulants reduces mortality in patients with severe sepsis. The long-term objectives of this proposal are to elucidate the mechanism by which coagulation proteases contribute to inflammation during sepsis. Our central hypothesis is that FXa activation of protease activated receptor 2 (PAR-2) enhances IL-6 expression and increases lethality during sepsis. We will also determine the role of thrombin (FIIa) -PAR- 1 signaling in sepsis. We will employ selective inhibitors of FXa and FIIa and analyze PAR-1- and PAR-2-dependent mice in a lethal mouse model of sepsis. In addition, we will use bone marrow transplantation to determine the role of TF and PAR-1 expression on monocytes versus
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endothelial cells in sepsis. Finally, we will generate mice that constitutively or inducibly express PAR-2 in endothelial cells to directly test our central hypothesis that PAR-2 is a key component of a pathogenic pathway involved in lethal sepsis. These studies should define the mechanism by which coagulation proteases enhance inflammation during sepsis. The clinical relevance of these studies is that they may provide new insight that can be used to develop improved therapeutic strategies for the treatment of patients with severe sepsis. Website: http://crisp.cit.nih.gov/crisp/Crisp_Query.Generate_Screen •
Project Title: REGULATION OF MACROPHAGE FUNCTION IN SURGICAL SEPSIS Principal Investigator & Institution: West, Michael A.; Professor; Surgery; Northwestern University Office of Sponsored Research Chicago, Il 60611 Timing: Fiscal Year 2002; Project Start 01-AUG-1990; Project End 31-JUL-2003 Summary: (adapted from the abstract.): Sepsis and multiple organ dysfunction syndrome (MODS) are the leading causes of mortality in critically ill surgical patients. TNF and IL-1 are pro-inflammatory cytokines produced by endotoxin- stimulated macrophages and are known to be important mediators in sepsis. The PI proposes that this regulated production of pro-inflammatory cytokines is a cardinal feature of sepsis leading to MODS. This is the result of sequential exposure to multiple inflammatory insults over time. The PI hypothesizes that a tolerant macrophage phenotype is the relevant macrophage response to inflammatory stimuli in patients with sepsis or MODS. Using an in vitro mouse macrophage model, they have previously demonstrated profound modifications in LPS-activated macrophage functions following LPS pretreatment. These LPS-tolerant alterations include: 1) an inhibition of TNF secretion; 2) augmentation of IL-1 secretion; 3) impaired activation of ERK1 and 2 and T38 mitogen activated kinase (MAPK); and 4) altered regulation of NF-kB. The present studies are designed to systematically investigate: 1) the mechanisms by which LPS pretreatment induces LPS tolerance; 2) the specific defects responsible for the altered response to LPS; and 3) to characterize the signal transduction alterations present in monocytes of patients with trauma, sepsis, or burns. The PI will focus on LPS signal transduction pathways that can be stimulated in the absence of serum or CD14, and do not require protein tyrosine kinases, MAPK, or NF-kB activation. They propose to extend previous investigations using mouse macrophages, human monocytic cell lines, and monocytes from clinical trauma patients, burn patients, or patients with sepsis. The previous results suggest that relatively early LPS activation signaling events depend on upstream MAPK activation and act by changes in both pre- and post-translational transcriptional processing. The PI will use specific agonists and antagonists to delineate the precise signal transduction alterations present in LPS-tolerant cells compared to naive macrophages. They hope that using monocytes from patients will help them to determine when and how alterations in signal transduction arise and how they can develop approaches to counteract or modulate macrophage-regulated cytokine production. Website: http://crisp.cit.nih.gov/crisp/Crisp_Query.Generate_Screen
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Project Title: REGULATION OF PROTEIN TURNOVER IN SEPSIS Principal Investigator & Institution: Vary, Thomas C.; Professor; Cellular/Molecular Physiology; Pennsylvania State Univ Hershey Med Ctr 500 University Drive Hershey, Pa 170332390
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Timing: Fiscal Year 2002; Project Start 01-JUL-1989; Project End 31-JUL-2003 Summary: The objectives of the studies described herein are to identify the loci responsible for the inhibition of protein synthesis in skeletal muscle during sepsis and to establish the mechanism(s) by which the inhibition can be reversed in order to develop treatment strategies to combat the severe muscle wasting associated with the septic process. Sustained muscle wasting contributes to the morbidity and mortality associated with sepsis. The defect in protein synthesis is localized to an impaired translation of mRNA at the level of peptide-chain initiation. Translation initiation is regulated at two steps: formation of the 43S pre-initiation complex (controlled by eukaryotic initiation factor 2 (eIF2) and eIF2B); and the binding of mRNA to the 40S ribosome (controlled by elF4E). We have identified a decreased activity of eIF2B as one defect in peptide-chain initiation and have shown that the muscle content of eIF2B protein is diminished 40 percent by sepsis. Therefore, reduced expression of eIF2B appeared a likely cause of the sepsis-induced inhibition of peptide-chain initiation in muscles of septic rats. However, protein synthesis can be stimulated 2-fold by perfusion of muscles from septic rats with buffer containing either IGF-I or elevated concentrations of amino acids by accelerating peptide-chain initiation without increasing the muscle content of eIF2B. Thus, effects of a reduced eIF2B expression on protein synthesis can be overridden, but the mechanisms responsible remain unknown. The hypothesis to be tested is that altered regulation of eIF2B and/or eIF4E mediates the changes in protein synthesis in sepsis. The specific aims of the studies proposed for the next project period are: (1) to evaluate the role of altered phosphorylation of eIF2B activity in controlling translation initiation during sepsis; (2) to investigate the effect of sepsis on eIF4E by measuring the amount of eIF4E found in the inactive 4E-BPI eIF4E complex and the active eIF4G eIF4E complex in muscle; (3) to investigate the mechanisms by which IGF-I stimulates translation initiation and contrast the response of skeletal muscle protein synthesis to IGF-I with that of insulin during sepsis; (4) to investigate the mechanisms by which amino acids stimulate translation initiation, and hence protein synthesis, during sepsis; and (5) to investigate the mechanisms by which chronic infusion of TNF or IL-1 cause an inhibition of protein synthesis in skeletal muscle. The research design will be to correlate changes in eukaryotic factor activity with rates of protein synthesis to establish which control mechanisms are important for regulating protein synthesis in skeletal muscle during sepsis. Website: http://crisp.cit.nih.gov/crisp/Crisp_Query.Generate_Screen •
Project Title: REGULATION OF THE (POLY) SIALIC VIRULENCE FACTOR Principal Investigator & Institution: Vimr, Eric R.; Associate Professor; Pathobiology; University of Illinois Urbana-Champaign Henry Administration Bldg Champaign, Il 61820 Timing: Fiscal Year 2004; Project Start 01-JUL-1998; Project End 31-DEC-2008 Summary: (provided by applicant): Bacterial sepsis and meningitis are the leading causes of mortality and hospitalization by infectious agents in the United States. Sepsis alone accounts for >700,000 life-threatening infections each year and over 200,000 deaths, with economic losses greatly exceeding one billion annually. Extraintestinal pathogenic Escherichia coli (ExPEC) are the predominant agents of these infections, with one serotype, E. coli O18:K1:H7, emerging as the leading cause of gram-negative neonatal meningitis and the most prevalent isolate from uncomplicated cystitis in women, thus making it a useful model of ExPEC infections in general. How ExPEC breach the host's mucosa and reach the systemic concentration (bacteremia) necessary for triggering sepsis and meningitis is not understood, although cell-surface
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modification with sialic acid is an important determinant. In this continuation application, we propose to complete our studies to determine how the sialic and polysialic acid virulence factors are synthesized, and then to identify other gene products required for systemic disease using a functional genomic approach. Specific Aim 1 will complete the analysis of sialic acid biosynthesis and the regulation of sialate metabolism. Preliminary data indicate that the synthesis of N-acetylmannosamine (ManNAc), the first committed step in sialate biosynthesis, does not involve ManNAc 6phosphate. X-ray crystallography and NMR spectroscopy will complement the objectives of this aim. In Specific Aim 2, the structural basis for PSA biosynthesis will be determined by analyzing chimeric polysialyltransferases (polySTs) and using sitedirected mutagenesis to understand catalytic mechanism. Our preliminary data indicate that these important enzymes are structurally and functionally distinct from mammalian polySTs, suggesting wide therapeutic potential for specific polyST inhibitors. Specific Aim 3 will apply signature-tagged mutagenesis to identify disease traits that are either distinct from PSA or function in concert with this capsule to define ExPEC virulence. Our objective in this aim is to establish the minimal systemic ExPEC "pathotype". The proposed studies have direct application to the development of new therapeutic approaches by identifying novel targets for non-antibiotic drug or vaccine design. New approaches are urgently needed to prevent or treat ExPEC infections in the rapidly aging US population, the very young and the increasingly large number of immunocompromised patients. Website: http://crisp.cit.nih.gov/crisp/Crisp_Query.Generate_Screen •
Project Title: REGULATION OF THE HUMAN INOS GENE IN SEPSIS AND TRAUMA Principal Investigator & Institution: Geller, David A.; Surgery; University of Pittsburgh at Pittsburgh 350 Thackeray Hall Pittsburgh, Pa 15260 Timing: Fiscal Year 2002; Project Start 01-JUN-1995; Project End 31-MAY-2004 Summary: Sepsis, trauma, and infection initiate systemic responses regulated by cytokines and other inflammatory mediators. In extreme conditions, this response can progress to multiple organ failure, a major cause of mortality in surgical patients. Advances in our understanding of this syndrome are based on the discovery that the inducible nitric oxide synthase (iNOS) gene is expressed in nearly every organ and tissue during the septic response. While NO synthesis has beneficial effects during acute inflammation, over-production of NO during sepsis can be detrimental with massive vasodilation and hypotension. Chronic expression of iNOS has been implicated in NOmediated cytotoxicity leading to diabetes, arthritis, neurode generative disorders, and certain cancers. Our laboratory has cloned the human iNOS gene from cytokinestimulated human hepatocytes. We then isolated the promoter region of the human iNOS gene and have shown that the cytokine-responsive DNA elements are located upstream from -4.7 kb. Recently we have characterized a novel NF-kappaB enhancer region that regulates iNOS transcription in response to TNFalpha or IL-1beta, and have shown a role for STAT1alpha in mediating IFNgamma induction. We have also identified extracellular signals that down-regulate iNOS expression including p53 tumor suppressor protein, steroids, heat shock, certain growth factors, and NO itself. Since the human iNOS gene is tightly regulated, we hypothesize that several mechanisms are working jointly to control the expression of this gene. We predict that this will include cytokine-stimulated nuclear factors that exert either positive or negative control over transcription, as well as post-transcriptional mechanisms that regulate iNOS mRNA stability and translational efficiency. In this proposal, we will pursue two interrelated
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specific aims. AIM I: TO DEFINE THE TRANSCRIPTIONAL MECHANISMS AND FUNCTIONAL PROMOTER ELEMENTS RESPONSIBLE FOR INDUCTION AND SUPPRESSION OF THE HUMAN iNOS GENE. Additional promoter regions will be fully sequenced and characterized in transfection experiments. DNA elements will be analyzed that are required for cytokine- responsiveness, with an initial focus on the interactions between NF-kappaB and STAT1alpha. Specific mechanisms for gene suppression by p53, steroids, heat shock, TGF-beta, and NO will be sought. Positive and negative transcription will be identified by gel shifts and in vivo footprinting assays. AIM II. TO DETERMINE THE POST-TRANSCRIPTIONAL MECHANISMS INVOLVED IN REGULATION OF THE HUMAN iNOS GENE. Post-transcriptional changes in mRNA stability or translational efficiency can also regulate gene expression. Cytokines will be tested for effects on human iNOS mRNA stability. Changes in translational efficiency in response to these agents will be measured by pulse-chase experiments. The 3'-untranslated region of the human iNOS gene will be analyzed for elements that mediate these effects. At the completion of our studies, we will have characterized the molecular regulation of the human iNOS gene. The information gained will increase our understanding of the control of iNOS transcription, describe novel mechanisms of cytokine-synergy in signal transduction, and help in designing therapeutic strategies for pathophysiological disease states where cytokine expression is relevant. Website: http://crisp.cit.nih.gov/crisp/Crisp_Query.Generate_Screen •
Project Title: ROLE OF S-NITROSOHEMOGLOBIN IN SEPSIS Principal Investigator & Institution: Patel, Rakesh P.; Pathology; University of Alabama at Birmingham Uab Station Birmingham, Al 35294 Timing: Fiscal Year 2002; Project Start 01-APR-2002; Project End 31-MAR-2006 Summary: (provided by applicant): Septic shock is a leading cause of mortality in hospitalized patients and is defined as the systemic inflammatory response to infection. This disease is characterized by hypotension and inflammatory damage to organs including the lung and which ultimately leads to multiple organ failure and death. Nitric oxide (NO) has a critical role in this disease with NO produced from iNOS receiving much attention. However, the role of NO in sepsis remains unclear with both detrimental and beneficial roles reported. An important regulator of NO function in the vasculature is Hemoglobin (Hb). Recent insights into the reactions between NO and Hb have shown that in addition to reactions with the heme, NO also interacts with a critical cysteine residue forming S-nitrosohemoglobin (SNOHb). SNOHb can elicit NO dependent effects and has been investigated principally in the context of physiological regulation of blood flow. The potential role of SNOHb in pathology has not been considered however. Recent studies show that SNOHb is elevated during endotoxemia and in this proposal, the novel concept that SNOHb mediates endotoxin induced hypotensive and inflammatory responses, by modulating production of different redox congeners of NO is put forward. The biological role of redox derivatives of NO, including nitroxyl anion (N0-) in disease remains largely unexplored although functions in promoting vasodilatation and stimulation of inflammatory responses have been suggested. Preliminary data presented herein suggest that SNOHb dependent vasorelaxation occurs via formation of N0- and vasorelaxing effects of red blood cells purified from endotoxin treated rats are demonstrated. These observations have led to the hypothesis that systemic hypotension and inflammation observed in sepsis are mediated by SNOHb. This hypothesis will be tested by pursuit of the following specific aims: 1) Determine the mechanism of SNOHb formation in endotoxic shock; 2) Determine the vasodilatory mechanisms of SNOHb; and 3) Investigate the role of
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SNOHb as a mediator of inflammatory damage in the lung. Accomplishment of these aims will yield novel insights into both the molecular mechanisms by which NO impacts upon the pathogenesis of septicemia and on possible therapeutic strategies to treat this inflammatory disease. Website: http://crisp.cit.nih.gov/crisp/Crisp_Query.Generate_Screen •
Project Title: ROLE OF T CELLS IN SELF-LIMITED MUCOSAL INFECTIONS Principal Investigator & Institution: Bry, Lynn; Brigham and Women's Hospital 75 Francis Street Boston, Ma 02115 Timing: Fiscal Year 2003; Project Start 01-AUG-2003; Project End 31-JUL-2007 Summary: (provided by applicant): We have found that the early adaptive serum immunoglobulin response plays an important role in surviving infection with noninvasive mucosal pathogens. B cells and CD4+ T cells are required to survive infection with Citrobacter rodentium, the mouse homolog for the enteropathogenic E. coli (EPEC). Wild-type mice develop small breaks in epithelial integrity during infection, allowing C. rodentium and members of the normal flora a direct portal of entry into the host. However, immunocompetent mice develop an early and robust serum IgM response and have few colony forming units (CFU) in end organs including liver and spleen. In contrast, infection proves lethal in mice lacking B cells or CD4+ T cells. Colonic infection leads to significant polymicrobial sepsis with damage to end organs. Unlike wild-type mice, CD4-deficient animals fail to mount pathogen-specific serum IgM or IgG responses during active infection. These results implicate a critical role for the systemic adaptive immune response in surviving and eventually clearing a mucosal infection. This research plan outlines a strategy to (1) establish the protective capacity of serum immunoglobulins during active infection, (2) establish the function and location(s) where CD4+ T cells stimulate a pathogen-specific humoral response in adoptively transferred CD4-deficient mice, (3) determine the location, immunophenotype, and cytokine secretion profiles of adoptively transferred CD4+ T cells, and (4) identify T cell co-stimulatory molecules and Th cytokines important in this response. In conjunction with these aims the candidate proposes a curriculum to further training in immunology, lymphocyte biology, pathology and ethical conduct in research. The candidate has completed the core clinical training as a resident in pathology and will devote at least 75% effort towards research. This training provides a necessary step in the candidate's goal to become an independent researcher investigating questions in mucosal immunology and host-microbial cross-talk in intestinal environments. Website: http://crisp.cit.nih.gov/crisp/Crisp_Query.Generate_Screen
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Project Title: ROLE OF TRAJ IN NEONATAL E. COLI SEPSIS AND MENINGITIS Principal Investigator & Institution: Badger, Julie L.; Children's Hospital Los Angeles 4650 Sunset Blvd Los Angeles, Ca 900276062 Timing: Fiscal Year 2003; Project Start 01-APR-2003; Project End 31-MAR-2008 Summary: (provided by applicant): Despite the use of modern therapeutics and diagnostic measures, neonatal bacterial sepsis and meningitis continues to be a disease with unacceptable rates of morbidity and mortality. The fatality rate of this disease is 1575 percent; furthermore, approximately 50 percent of meningitis survivors have significant neurological and developmental abnormalities. The primary, route of infection for E. coli K1 (the most common causative agent of gram-negative meningitis) is oral. Following intestinal colonization, bacteria translocate through the GI tract to
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extra-intestinal sites of mesenteric lymph nodes (MLN), liver, spleen, and the blood. E. coli K1 then multiply systemically within the bloodstream reaching a necessary threshold of bacteremia to gain access to the central nervous system (CNS). Recently we identified an E. coli K1 plasmid-encoded blood-brain barrier (BBB) invasion gene, traJ, with homology to traJ of various F-like plasmid tra operons. Our preliminary data indicate that the traJ mutation specifically attenuates dissemination from the MLN to the liver, spleen, blood, and the CNS in the neonatal rat. In addition, although animals orally infected with the traJ mutant demonstrated a decrease or no recoverable bacteria in the liver or spleen, these tissues showed a significant inflammatory response. In vitro studies determined that the traJ mutant is taken-up less by macrophages and shows a loss of a 55 kDa-secreted protein. The central hypothesis of this application is that TraJ controls the expression of a set of genes whose products (i.e., 55 kDa secreted protein) are involved in E. coli K1 dissemination, systemic infection and crossing the BBB, and these events within the disease process occur via TraJ-dependent interactions with professional phagocytes. The following proposed experiments are designed to test and substantiate our hypotheses. We aim to 1) Elucidate the genetic and molecular characteristics of the traJ region and plasmid, evaluate the ability of the endogenous plasmid to self-transfer, and determine the potential role of the traJ-containing plasmid in E. coli K1 neonatal rat virulence, 2) Evaluate the function of the TraJ-regulated proteins (i.e., 55 kDa-secreted protein) in E. coli K1 systemic dissemination and meningitis, and 3) Determine the mechanism of TraJ-dependent host inflammatory response in the neonatal rat. Improved knowledge of molecular mechanisms for early systemic dissemination and the exact interplay of the host inflammatory response during these events will assist in achieving our long-term goal of identifying novel rational approaches to development of new treatments and preventive strategies for E. coli K1 sepsis and meningitis. Website: http://crisp.cit.nih.gov/crisp/Crisp_Query.Generate_Screen •
Project Title: SALT AND WATER TRANSPORT IN THE ALCOHOLIC LUNG Principal Investigator & Institution: Eaton, Douglas C.; Professor and Deputy Chairman of Physiol; Emory University 1784 North Decatur Road Atlanta, Ga 30322 Timing: Fiscal Year 2003; Project Start 01-FEB-2003; Project End 31-DEC-2007 Summary: (provided by applicant): Salt and water transport by lung epithelial cells is critical for normal clearance of fluid in the developing and mature lungs. A delicate balance between alveolar fluid secretion and absorption results in a thin fluid layer on the surface of the airways that helps promote pulmonary gas exchange and mucociliary clearance of foreign particles from the lung. The alveolar epithelial barrier formed by lung epithelial cells and tight junctions between the cells play a key role in this process, and disruption of the barrier function can result in alveolar flooding. Chronic alcohol exposure appears to compromise the alveolar barrier. Nonetheless, compensatory increases in salt transport in the alcoholic lung appear to be sufficient to maintain approximately normal levels of airway surface fluid. However, alcoholic lungs when challenged by any significant stress (like major trauma or sepsis) are much more likely to develop edema implying that the salt and water transport mechanisms cannot respond to increased demand as nonalcoholic lungs can. It is hypothesized that alcoholinduced changes in epithelial barrier function and transport mechanisms predispose the lungs to acute edematous lung injury. While there is now substantial evidence that the maintenance of salt and water transport is a strongly regulated, energy-dependent process, the pathways for salt and water transport are not clearly defined in the normal lung, let alone how they are modified in the alcoholic lung. It does seem likely that some
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regulatory mechanism controlling the response of lung salt and water transport stress is abnormal in alcoholic lungs. It is hypothesized that abnormal glucocorticoid and TGFbeta responsiveness of lung epithelial cells prevents stress-induced increases in lung salt and water transport in alcoholic lungs.There are three specific aims of this proposal. The first aim is to determine if transport characteristics of the alcoholic lung are different from normal lung. The second aim is to determine how transport in stressed alcoholic lung differs from normal and alcoholic lung. The third aim to elucidate the cellular mechanisms responsible for alcohol-induced changes in lung transport. These experiments will improve our understanding of how chronic alcohol exposure alters alveolar fluid balance under normal and stressful conditions, and help in devising therapeutic strategies to prevent edematous lung injury. Website: http://crisp.cit.nih.gov/crisp/Crisp_Query.Generate_Screen •
Project Title: SENSITIVITY OF TCR SIGNALING TO GSH LOSS IN HIV DISEASE Principal Investigator & Institution: Herzenberg, Leonore A.; Genetics; Stanford University Stanford, Ca 94305 Timing: Fiscal Year 2004; Project Start 01-FEB-2004; Project End 31-JAN-2006 Summary: (provided by applicant): Previous work in our laboratory and elsewhere has established that intracellular redox status measured in terms of intracellular glutathione (GSH) levels regulates T cell function. Inflammatory responses such as the activation of NF-kappaB are up-regulated under oxidative stress (when GSH is depleted) whereas activation events required for T cell (TCR) signaling, including tyrosine kinase phosphorylations and calcium influx, are down-regulated under these conditions but favored when intracellular GSH levels are normal. Consistent with these findings, intracellular GSH tends to be depleted in a wide variety of diseases (e.g., HIV infection, sepsis, diabetes, neoplasia) in which T cell function is often impaired. To define the biochemical mechanisms that underlie the redox regulation of TCR signaling, we propose here to combine the 11-color Fluorescent Activated Cell Sorter (Hi-D FACS) technology that we have developed to measure intracellular redox status in human T cell subsets with Hi-D FACS methods to quantitate phosphorylation of individual kinases in these T cell subsets. By examining the functional consequences of altering redox status (depleting/repleting GSH) in freshly isolated T cells from healthy donors, and by probing the relationship between redox status and T cell function in patients with various diseases, we contribute significantly to basic and clinical understanding of the impact of GSH depletion on immune function. Further, since treatment with nontoxic cysteine pro-drugs can replenish GSH, our findings may open the way to introduction of adjunct therapies to increase immune surveillance and bolster responses to infection. Finally, the technology we develop here will provide new and better ways to monitor disease progress and evaluate therapeutic interventions. Website: http://crisp.cit.nih.gov/crisp/Crisp_Query.Generate_Screen
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Project Title: SYSTEMS APPROACH TO INNATE IMMUNITY-INFLAMMATIONSEPSIS Principal Investigator & Institution: Ulevitch, Richard J.; Chairman & Professor; Scripps Research Institute Tpc7 La Jolla, Ca 92037 Timing: Fiscal Year 2002; Project Start 30-SEP-2002; Project End 31-JUL-2007 Summary: In the United States, recent estimates suggest that more than 750,000 patients per year are at high risk for developing septic shock with mortality rates reaching 60%. Thus there are huge societal and financial costs associated with this syndrome. Because
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of its high incidence and poor prognosis, we propose basic research focused on the innate immune system and its interface with the adaptive immune system. We acknowledge that the pathophysiology of severe sepsis/shock is exceedingly complex. There is little doubt, however, that there is often a progression from infection --) normal systemic response (sepsis) --) severe sepsis --) shock and that the primary event in this sequence is infection. There is also important evidence that the intensity of the inflammatory process at the site(s) of infection determine the severity of the systemic reaction to infection, and thus the occurrence of severe sepsis and shock. It is our contention that the only means to approach such a complex system is to use appropriate cellular and animal model systems and apply the principles of systems biology in their analysis. To accomplish this we have brought together investigators with many different scientific backgrounds and specialties that include cutting-edge bioinformatics, genomic and proteomic analyses, cell biology, studies of innate and adaptive immunity, biological network modeling etc. Most importantly, we have combined experts in the field of innate immunity with "new scientific blood" such as experts in the fields of genomics, proteomics and bioinformatics. It is our contention that in the context of this GLUE GRANT this combination of investigators will, in our highly interactive and interdependent setting, substantially expand the formulation of new concepts for this field. This effort will form the basis for a great many hypothesis-driven studies by the scientific community at-large, both basic and clinical. These studies will also identify potential new drug targets that could lead to new therapies for sepsis and shock. This would directly impact the clinical outcome of these critically ill patients. Website: http://crisp.cit.nih.gov/crisp/Crisp_Query.Generate_Screen •
Project Title: TARGETED INTERLEUKIN-10 GENE THERAPY IN SEPSIS SYNDROMES Principal Investigator & Institution: Moldawer, Lyle L.; Professor; Surgery; University of Florida Gainesville, Fl 32611 Timing: Fiscal Year 2002; Project Start 01-APR-2001; Project End 31-MAR-2005 Summary: (Verbatim from the applicant's abstract) The treatment of the hospitalized patient with sepsis remains a clinical conundrum, due in large part to the complexity of the host response to infection and tissue injury. Pro- and antiinflammatory cytokines play critical roles in the development of sepsis syndromes, but their production is often localized to individual tissues, rather than the systemic circulation. Current therapeutic approaches for the treatment of sepsis syndromes involve the systemic administration of biological response modifiers, such as cytokine antagonists or antiinflammatory agents. To date, none of these agents has proven effective, and one explanation may be that their systemic administration is an inefficient or even inappropriate means to treat a more localized inflammatory or immunosuppressive response. We have proposed that he targeted delivery of protein-based therapies using a recombinant adenoviral vector can directly modulate the inflammatory or apoptotic processes occurring in the target tissue, without systemic immune suppression or activation. Therefore, the overall objective of these studies is to determine the feasibility of adenovirus gene transfer as a herapeutic modality. More specifically, the goals of this proposal are to determine whether the forced overexpression of initially IL-10, but also in later years, a dominant negative I-KB super-repressor, will blunt the sepsis responses in arget organs without producing systemic immune suppression. The two specific aims of the current application are: 1) To determine whether targeted expression of IL- 10 in the lungs of mice with an adenoviral vector can prevent the development of respiratory distress syndrome in a zymosan-induced model of multisystem organ failure, without increased
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expression in other organs or the induction of T-cell hyporesponsiveness and immune suppression, and 2) To determine the mechanisms by which targeted expression of IL10 in the thymus of mice can reduce caspase-3 iependent thymocyte apoptosis and improve outcome in a cecal ligation and puncture model of a compensatory mtiinflammatory response syndrome. The studies proposed in this application will ultimately determine the utility and safety ofadenovirus based gene therapy as a means of targeting protein-based therapies to individual tissues and organs in sepsis syndromes. Website: http://crisp.cit.nih.gov/crisp/Crisp_Query.Generate_Screen •
Project Title: THE LPS RESPONSIVENESS OF TLR2 AN TLR4 IN THE NEUTROPHIL Principal Investigator & Institution: Arndt, Patrick G.; Medicine; University of Colorado Hlth Sciences Ctr P.O. Box 6508, Grants and Contracts Aurora, Co 800450508 Timing: Fiscal Year 2002; Project Start 15-SEP-2001; Project End 31-JUL-2006 Summary: (provided by applicant) The sepsis syndrome and sepsis induced Acute Respiratory Distress Syndrome (ARDS), associated with expo- sure to LPS, are important clinical entities without available specific therapies. The recent identification of the Toll- like receptors (TLRs), in particular TLR2 and TLR4 as LPS receptors has advanced our understanding of the initiation of signaling after LPS exposure. We hypothesize that TLR4 is the predominant receptor responsible for LPS induced NF-KB and/or p38 activation in the neutrophil with the signaling pathway involving IRAK 2 and M, syk and Rac2. Although, alternatively, LPS signaling may occur through TLR2 with less avidity and with involvement of other IRAK sub-species, tyrosine kinases, or small G proteins. We show here that human neutro- phils and PLB-985 cells express mRNA for TLRI-6 and express TLR2 protein. In addition, we show here in PLB-985 cells, that IRAK, the tyrosine kinase syk, and the small protein Rac2 associate with TLR2 at baseline and after LPS exposure, suggesting their involvement in LPS signaling through TLR2. We propose to investigate in neutrophils, both human and murine, and the PLB985 cell line: 1. the role of TLR2 and TLR4 in NF-KB and p38 activation, including their interdependence, 2. the macromolecular complex which associate with TLR2 or TLR4 after LPS exposure, and 3. the role and activation of IRAK 2 & M, the tyrosine kinases syk and lyn, and the small G proteins Rac2 and Cdc42 in LPS signaling. To accomplish these goals, we will develop inducible antisense retroviral techniques for the creation of antisense TLR2, TLR4, and IRAK ex- pressing cell lines, dominant negatives for TLR2, TLR4, and IRAK M, and novel immunoprecipitation techniques for 2D gel electrophoresis to examine the macromolecular complexes associated with TLR2, TLR4, and the IRAK subspecies. I will also develop techniques in 2D gel electrophoresis and protein identification by mass spec- trometry which will benefit future investigations into signaling pathways. An improved understanding of the recognition of LPS, and the signaling pathways initiated by LPS, in the neu- trophil are important to improve the understanding of the underlying pathophysiology of sepsis syndrome and sepsis induced ARDS. Website: http://crisp.cit.nih.gov/crisp/Crisp_Query.Generate_Screen
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Project Title: THE ROLE OF MICROBIAL ORGANISMS IN ACUTE LUNG INJURY Principal Investigator & Institution: Doerschug, Kevin C.; Cardiovascular Center; University of Iowa Iowa City, Ia 52242 Timing: Fiscal Year 2003; Project Start 01-APR-2003; Project End 31-MAR-2008
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Summary: This application is to support the development of Dr. Kevin Doerschug so that he will become an accomplished clinician-scientist and independent researcher. Dr. Gary Hunninghake will assume responsibility as mentor to ensure the success of the career development plan. The focus of the proposal is an intensive research experience in studies of human lung injury and sepsis. To achieve this, Dr. Doerschug will continue formal training in the design and analysis of human studies, as well as scientific techniques to analyze specimens from human patients with acute lung injury and sepsis. Dr. Doerschug will complete didactic training in these areas including classes in epidemiology, statistics, cell biology, and microbiology. Dr. Doerschug's classes will culminate in a Master's Degree in Translational Biomedical Research. The research mentorship will involve active development of and participation in studies designed to identify factors involved in the pathogenesis of acute lung injury and sepsis. Specifically, Dr. Doerschug will focus on the role of circulating microorganisms as the stimulus for the inflammation that characterizes these diseases. Initial studies will include identifying bacteremia or fungemia in patients with SIRS (a common risk factor for sepsis and acute lung injury), using PCR to detect previously undetectable organisms. Additionally, Dr. Doerschug will investigate the relationships between circulating microorganisms and the development of acute lung injury, focusing on patient physiology and outcomes. Once completed, these preliminary studies will more accurately describe the relationships between infection and inflammation in these disorders. Further studies can then focus on the modification of host responses to these infections. Throughout Dr. Doerschug's career development, he will interface with trainees in clinical research, pulmonary medicine, and microbiology. This intellectually rich environment will help foster the career development of Dr. Doerschug, providing ample opportunity for critical review throughout the period of this award. Both the mentor and the institution are highly committed to Dr. Doerschug's scientific development and academic success. Website: http://crisp.cit.nih.gov/crisp/Crisp_Query.Generate_Screen •
Project Title: THE ROLE OF TISSUE FACTOR IN TRAUMA Principal Investigator & Institution: Armstead, Valerie E.; Anesthesiology; Thomas Jefferson University Office of Research Administration Philadelphia, Pa 191075587 Timing: Fiscal Year 2002; Project Start 01-FEB-2000; Project End 31-JAN-2005 Summary: (Adapted from the applicant's abstract) Trauma is a serious health problem in the United States and the leading cause of death in young people. Trauma causes tissue injury, resulting in disruption of normal barriers, leading to dysfunction of physiologic systems and organs. Significant damage to the vasculature results in abnormalities in blood flow and coagulation that further compromise vital organ function. Tissue factor (TF) is a transmembrane glycoprotein that is normally expressed in epithelial cells, glial cells, mucosal cells and adventitia. TF binds to factor VII and active VIIa to trigger the extrinsic and intrinsic coagulation pathways. Activation of TF expression in cells where it is normally not expressed, such as endothelium, results in thrombosis and coagulation abnormalities. Inhibition of TF activity has been shown to increase survival in animal models of sepsis. Inappropriate activity of the coagulation cascade during trauma may be, in part due, to a potent activator such as TF. The proposed research focuses on studying the expression and activity of TF in trauma. The major hypothesis to be tested is that TF expression is an important mechanism of endothelial dysfunction and tissue damage in trauma. The specific aims of this proposal involve four areas of investigation. The first aim is to characterize TF gene expression following trauma. This goal will be accomplished by utilizing a well-recognized model of soft tissue trauma and murine
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traumatic shock (Noble-Collip drum trauma). After soft tissue trauma is induced in anesthetized rats, vital signs will be monitored and TF expression measured in vital organ mRNA, tissue extracts, and plasma. Coagulation activity will be measured with standard assays. The second aim will be to determine the relationship between TF and two other important mediators of leukocyte-endothelial-platelet interaction, P-selectin and nitric oxide (NO). This goal will be achieved by using Noble-Collip drum trauma, as well as cultured monocytes, macrophages, and vascular cells, and determining TF expression while manipulating experimental conditions with NO and P-selectin agonists and antagonists. The third aim will be to investigate the effect of inhibition of TF activity in traumatic shock by administering tissue factor pathway inhibitor, TF antibody and inhibitors of early activation of TF gene transcription. The fourth aim is to analyze the regulatory mechanisms of the induction of TF gene expression related to trauma by administering analogs of potential TF regulatory factors to our experimental models. The long-term objective is to determine if modulation of TF subsequent to trauma changes the natural course of traumatic shock. Website: http://crisp.cit.nih.gov/crisp/Crisp_Query.Generate_Screen •
Project Title: THERAPEUTIC USE OF INTER-ALPHA INHIBITOR IN SEPSIS Principal Investigator & Institution: Lim, Yow-Pin; Prothera Biologics, Llc 87 Walmer Ave East Providence, Ri 02914 Timing: Fiscal Year 2002; Project Start 01-MAY-2002; Project End 31-MAR-2003 Summary: The primary goal of this proposed research is targeted towards demonstrating the commercial utilization of human plasma derived inter- alpha inhibitor proteins (IaIp) an effective therapy of sepsis. IaIp are serine protease inhibitors found normally at high levels in human plasma. The fact that no person with complete absence of IaIp has ever been detected suggests an essential physiological role for these proteins. In patients with severe sepsis, the plasma level of IaIp decrease significantly and the decrease correlates with mortality. We propose that administration of IaIp to restore the imbalance between these natural protective inhibitors and destructive proteases will prevent organ injuries and ultimately reduce sepsis related mortality. Our preliminary animal studies in a sepsis rat model of cecal ligation and puncture (CLP) have demonstrated the beneficial effects of IaIp in maintaining hemodynamic stability, preventing organ injury, and improving survival during sepsis. In this study, we propose to optimize a protocol for purification of IaIp from human crude plasma and to evaluate and confirm the beneficial effects of the purified IaIp in the CLP rat model. The ultimate goal of these studies is to develop a novel plasma derived therapeutic that is safe and effective for sepsis. PROPOSED COMMERCIAL APPLICATIONS: There is a serious unmet medical need for more than 1.5 million people worldwide annually who suffer from sepsis. Since there are currently no agents effective for sepsis, treatment is limited to antibiotic and supportive therapy. The magnitude of the need for an effective novel therapeutic agent in reducing sepsis mortality and morbidity is immense and it creates an enormous market potential estimated at $10-25 billion/year in the US alone. Inter-alpha inhibitor proteins appear to be an effective therapeutic agent of sepsis. The proposed study is expected to lead to the development of novel human plasma derived protein that is safe and effective for sepsis. Website: http://crisp.cit.nih.gov/crisp/Crisp_Query.Generate_Screen
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Project Title: THERMAL INJURY INDUCED ALTERATIONS IN IMMUNE FUNCTION Principal Investigator & Institution: Schwacha, Martin G.; Assistant Professor of Surgery; Surgery; University of Alabama at Birmingham Uab Station Birmingham, Al 35294 Timing: Fiscal Year 2002; Project Start 01-JUL-1999; Project End 31-JUL-2004 Summary: Immune dysfunction, susceptibility to sepsis and multiple organ failure are complications associated with thermal injury. Evidence suggests that activation of a proinflammatory cascade after burn injury plays an important role in their development. With regards to this, macrophages (Mphi) are major producers of pro-inflammatory mediators with increased productive capacity being observed post-burn. Thus, Mphi hyperactivity (as defined by increased productive capacity for pro-inflammatory mediators) may be of fundamental importance in the development of these complications. Nonetheless, the precise mechanisms responsible for the alterations in Mphi activity are unclear. We have utilized a murine scald burn model (3rd degree, 25 percent total body surface area) and our preliminary results indicated that at 4-7 days post-burn Mphi were "hyperactive" as increased productive capacity for nitric oxide, TNF-alpha, IL-6 and PGE2 was observed. Furthermore, at 4-7 days post-burn, but not earlier, Mphi hyperactivity was responsible for the suppression of T cell function and at 7 days post-burn mice were significantly more susceptible to the lethal effects of sepsis. With regards to T cells, we have observed significant mortality (approximately 75 percent) during the initial 48 hr. post-burn period in mice lacking gamma/delta T cells (gamma/delta T cell knock-out mice) and Mphi isolated from surviving mice at 7 days post-burn appear not to be "hyperactive". These findings suggest a dual role for gamma/delta T cells in burn injury pathogenesis; 1) survival early and; 2) induction of Mphi hyperactivity later. Our preliminary results suggest that Mphi hyperactivity postburn is related to alterations in sensitivity to CAMP, however, the mediators and mechanisms responsible for Mphi hyperactivity post-burn and its role in immune dysfunction remain to elucidated. Moreover, the impact of burn excision, a common clinical practice, on Mphi hyperactivity and susceptibility to sepsis post-burn is unknown. It is our hypothesis that Mphi hyperactivity post-burn is mediated by gamma/delta T cells and altered cAMP responses leading to the development of immune dysfunction. Therefore, we propose to determine the following: 1) The relationship between gamma/delta T cells, Mphi and survival early (initial 48 hr.) postburn; 2) The role of gamma/delta T cells in the induction of Mphi hyperactivity late (7 days) post-burn; 3) The mechanisms responsible for Mphi hyperactivity post-burn; and 4) The effect of burn wound excision on Mphi hyperactivity and increased susceptibility to sepsis. A more comprehensive understanding of the relationship between Mphi activity, T cell function, and the development of immune dysfunction following thermal injury should hopefully provide the basis for improved therapeutic regimes in the treatment of burn patients. Website: http://crisp.cit.nih.gov/crisp/Crisp_Query.Generate_Screen
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Project Title: THROMBIN, SEPSIS AND MECHANISMS OF INFLAMMATION Principal Investigator & Institution: Tiruppathi, Chinnaswamy; Associate Professor; Pharmacology; University of Illinois at Chicago 1737 West Polk Street Chicago, Il 60612 Timing: Fiscal Year 2003; Project Start 01-FEB-1999; Project End 31-JAN-2007 Summary: (provided by applicant): The overall objective of this renewal application is to test the hypothesis that the activation of Trp channel-mediated Ca2+ entry in
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endothelial cells increases lung microvascular permeability and promotes lung edema. We have shown that thrombin activation of Proteinase-Activated Receptor-1 (PAR-1) expressed on endothelial cell surface induces an increase in intracellular Ca2+, which is critical in the mechanism of increased endothelial permeability. The increase in intracellular Ca2+ concentration ([Ca2+]i) is dependent on both Ca2+ store depletion and the Ca2+ store depletion-mediated Ca2+ influx. We showed that the prevention of Ca2+ influx drastically reduced the thrombin-induced increase in endothelial permeability. We also show that the Ca2+ influx due to store depletion occurs through activation of the transient receptor potential (Trp) gene family of channels expressed in endothelial cells. Our supporting data show that Trp1 (Trp gene homologue) is predominantly expressed in human endothelial cells and increased expression of Trp1 augments the Ca2+ influx in endothelial cells. Further, we show that Src kinase activation plays an important role in the regulation of Ca2+ influx via Trp channels. Deletion of Trp4 gene (predominant Trp isoform expressed in mouse endothelial cells) in mouse impairs thrombin-induced increase in lung microvascular permeability. The association of Trp1 with caveolin-1 (the principal protein of caveolae) in response to thrombin is required for the Ca2+ influx in endothelial cells. We also show that the inflammatory cytokine, tumor necrosis factor-alpha (TNFa) increases Trp1 expression in endothelial cells. Based on these supporting data, we propose to (i) determine the role of Src kinase signaling in the mechanism of Ca2+ influx in endothelial cells via Trp channels (ii) study the role of caveolin-1 in the mechanism of Ca2+ influx through Trp channels and its role in the mechanism of increased endothelial permeability, and (iii) investigate the effects of inflammatory mediators, TNFa and lipopolysaccharide, on the functional expression of Trp channels and its role in the mechanism of increase in endothelial permeability. The proposed studies will utilize cell and in vivo approaches involving Src null (Src -/-), caveolin-1 null (cav1 -/-), and Trp4 null (Trp4 -/-) mice to address the role of Trp channels in activating Ca2+ influx in endothelial cells and in the mechanism of increased endothelial permeability. With the achievement of these studies, we will be able to provide new molecular insight into understanding the mechanism of vascular injury and tissue inflammation associated with states of hypercoagulation such as with sepsis. Website: http://crisp.cit.nih.gov/crisp/Crisp_Query.Generate_Screen •
Project Title: TREATMENT FOR GBS COLONIZATION IN PREGNANT WOMEN Principal Investigator & Institution: Bhatt, Bakul M.; Biomedical Development Corporation 500 Sandau, Ste 200 San Antonio, Tx 78216 Timing: Fiscal Year 2001; Project Start 20-SEP-1997; Project End 31-MAY-2004 Summary: (Adapted from Applicant's Abstract): The goal of this project is to further develop Iocide, a novel iodine-based antimicrobial technology, for use as a topical bacteriocide for group B streptococci (GBS) in the vaginal tract of pregnant women during the intrapartum period. GBS is the leading cause of neonatal bacterial disease and death, and most frequently occurs in neonates as sepsis, meningitis, and pneumonia. Despite clinical advances to date, GBS continues to be a major cause of illness and death among newborns. Due to the limitations of current clinical strategies, a need exists for alternate approaches to prevent GBS-related neonatal disease. The following specific aims have been designed to achieve this goal: 1) to optimize the formulation of the Iocide vaginal gel, 2) to determine the effective range of Iocide in vitro, 3) to demonstrate safety and efficacy to the Iocide in animals, and 4) to determine safety and efficacy of Iocide in human clinical trials. PROPOSED COMMERCIAL APPLICATION: NOT AVAILABLE
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Website: http://crisp.cit.nih.gov/crisp/Crisp_Query.Generate_Screen
E-Journals: PubMed Central3 PubMed Central (PMC) is a digital archive of life sciences journal literature developed and managed by the National Center for Biotechnology Information (NCBI) at the U.S. National Library of Medicine (NLM).4 Access to this growing archive of e-journals is free and unrestricted.5 To search, go to http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=Pmc, and type “sepsis” (or synonyms) into the search box. This search gives you access to full-text articles. The following is a sample of items found for sepsis in the PubMed Central database: •
A Double-Blind Placebo-Controlled Study of an Infusion of Lexipafant (PlateletActivating Factor Receptor Antagonist) in Patients with Severe Sepsis. by Suputtamongkol Y, Intaranongpai S, Smith MD, Angus B, Chaowagul W, Permpikul C, Simpson JA, Leelarasamee A, Curtis L, White NJ.; 2000 Mar; http://www.pubmedcentral.gov/articlerender.fcgi?tool=pmcentrez&artid=89748
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A Model of Infected Burn Wounds Using Escherichia coli O18:K1:H7 for the Study of Gram-Negative Bacteremia and Sepsis. by Busch NA, Zanzot EM, Loiselle PM, Carter EA, Allaire JE, Yarmush ML, Warren HS.; 2000 Jun; http://www.pubmedcentral.gov/articlerender.fcgi?tool=pmcentrez&artid=97598
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Actinobacillus equuli Septicemia: an Unusual Zoonotic Infection. by Ashhurst-Smith C, Norton R, Thoreau W, Peel MM.; 1998 Sep; http://www.pubmedcentral.gov/articlerender.fcgi?tool=pmcentrez&artid=105212
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Activity of lipopolysaccharide-binding protein-bactericidal/permeability-increasing protein fusion peptide in an experimental model of Pseudomonas sepsis. by Opal SM, Palardy JE, Jhung JW, Donsky C, Romulo RL, Parejo N, Marra MN.; 1995 Dec; http://www.pubmedcentral.gov/articlerender.fcgi?tool=pmcentrez&rendertype=abstr act&artid=163038
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Application of 16S rRNA Gene Sequencing To Identify Bordetella hinzii as the Causative Agent of Fatal Septicemia. by Kattar MM, Chavez JF, Limaye AP, Rassoulian-Barrett SL, Yarfitz SL, Carlson LC, Houze Y, Swanzy S, Wood BL, Cookson BT.; 2000 Feb; http://www.pubmedcentral.gov/articlerender.fcgi?tool=pmcentrez&artid=86205
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Assessment of a new hub design and the semiquantitative catheter culture method using an in vivo experimental model of catheter sepsis. by Segura M, Alia C, Valverde J, Franch G, Torres Rodriguez JM, Sitges-Serra A.; 1990 Nov; http://www.pubmedcentral.gov/picrender.fcgi?tool=pmcentrez&action=stream&blobt ype=pdf&artid=268222
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Assessment of Neutrophil Function in Patients with Septic Shock: Comparison of Methods. by Wenisch C, Fladerer P, Patruta S, Krause R, Horl W.; 2001 Jan; http://www.pubmedcentral.gov/articlerender.fcgi?tool=pmcentrez&artid=96030
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Adapted from the National Library of Medicine: http://www.pubmedcentral.nih.gov/about/intro.html.
With PubMed Central, NCBI is taking the lead in preservation and maintenance of open access to electronic literature, just as NLM has done for decades with printed biomedical literature. PubMed Central aims to become a world-class library of the digital age. 5 The value of PubMed Central, in addition to its role as an archive, lies in the availability of data from diverse sources stored in a common format in a single repository. Many journals already have online publishing operations, and there is a growing tendency to publish material online only, to the exclusion of print.
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Bacteriological and Serological Findings in a Further Case of Transfusion-Mediated Yersinia enterocolitica Sepsis. by Strobel E, Heesemann J, Mayer G, Peters J, MullerWeihrich S, Emmerling P.; 2000 Jul; http://www.pubmedcentral.gov/articlerender.fcgi?tool=pmcentrez&artid=87034
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Balance of inflammatory cytokines related to severity and mortality of murine sepsis. by Walley KR, Lukacs NW, Standiford TJ, Strieter RM, Kunkel SL.; 1996 Nov; http://www.pubmedcentral.gov/articlerender.fcgi?tool=pmcentrez&rendertype=abstr act&artid=174439
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Bench-to-bedside review: Toll-like receptors and their role in septic shock. by Opal SM, Huber CE.; 2002; http://www.pubmedcentral.gov/articlerender.fcgi?tool=pmcentrez&artid=137294
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Blood Cultures Positive for Coagulase-Negative Staphylococci: Antisepsis, Pseudobacteremia, and Therapy of Patients. by Souvenir D, Anderson DE Jr, Palpant S, Mroch H, Askin S, Anderson J, Claridge J, Eiland J, Malone C, Garrison MW, Watson P, Campbell DM.; 1998 Jul; http://www.pubmedcentral.gov/articlerender.fcgi?tool=pmcentrez&artid=104953
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Brevibacterium casei Sepsis in an 18-Year-Old Female with AIDS. by Brazzola P, Zbinden R, Rudin C, Schaad UB, Heininger U.; 2000 Sep; http://www.pubmedcentral.gov/articlerender.fcgi?tool=pmcentrez&artid=87423
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Case of Sepsis Caused by Bifidobacterium longum. by Ha GY, Yang CH, Kim H, Chong Y.; 1999 Apr; http://www.pubmedcentral.gov/articlerender.fcgi?tool=pmcentrez&artid=88684
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Catheter Sepsis Due to Mycobacterium chelonae. by Wallace RJ Jr, Brown BA.; 1998 Nov; http://www.pubmedcentral.gov/articlerender.fcgi?tool=pmcentrez&artid=105357
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Catheter-Related Sepsis Due to Rhodotorula glutinis. by Hsueh PR, Teng LJ, Ho SW, Luh KT.; 2003 Feb; http://www.pubmedcentral.gov/articlerender.fcgi?tool=pmcentrez&artid=149691
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CD40 Contributes to Lethality in Acute Sepsis: In Vivo Role for CD40 in Innate Immunity. by Gold JA, Parsey M, Hoshino Y, Hoshino S, Nolan A, Yee H, Tse DB, Weiden MD.; 2003 Jun; http://www.pubmedcentral.gov/articlerender.fcgi?tool=pmcentrez&artid=155725
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Central Line Sepsis in a Child Due to a Previously Unidentified Mycobacterium. by Hogg GG, Schinsky MF, McNeil MM, Lasker BA, Silcox VA, Brown JM.; 1999 Apr; http://www.pubmedcentral.gov/articlerender.fcgi?tool=pmcentrez&artid=88674
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Chemokine C10 Promotes Disease Resolution and Survival in an Experimental Model of Bacterial Sepsis. by Steinhauser ML, Hogaboam CM, Matsukawa A, Lukacs NW, Strieter RM, Kunkel SL.; 2000 Nov; http://www.pubmedcentral.gov/articlerender.fcgi?tool=pmcentrez&artid=97687
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Circulating complement proteins in patients with sepsis or systemic inflammatory response syndrome. by Stove S, Welte T, Wagner TO, Kola A, Klos A, Bautsch W, Kohl J.; 1996 Mar; http://www.pubmedcentral.gov/articlerender.fcgi?tool=pmcentrez&rendertype=abstr act&artid=170271
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Clinical correlations of serial quantitative blood cultures determined by lysiscentrifugation in patients with persistent septicemia. by Whimbey E, Wong B, Kiehn TE, Armstrong D.; 1984 Jun; http://www.pubmedcentral.gov/picrender.fcgi?tool=pmcentrez&action=stream&blobt ype=pdf&artid=271182
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Complement Activation in Relation to Capillary Leakage in Children with Septic Shock and Purpura. by Hazelzet JA, de Groot R, van Mierlo G, Joosten KF, van der Voort E, Eerenberg A, Suur MH, Hop WC, Hack CE.; 1998 Nov; http://www.pubmedcentral.gov/articlerender.fcgi?tool=pmcentrez&artid=108669
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Concordance of endotoxemia with gram-negative bacteremia in patients with gramnegative sepsis: a meta-analysis. by Hurley JC.; 1994 Sep; http://www.pubmedcentral.gov/picrender.fcgi?tool=pmcentrez&action=stream&blobt ype=pdf&artid=263953
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Critical advances in septicemia and septic shock. by Das UN.; 2000; http://www.pubmedcentral.gov/articlerender.fcgi?tool=pmcentrez&artid=137258
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Critical Role of CD14 for Production of Proinflammatory Cytokines and Cytokine Inhibitors during Sepsis with Failure To Alter Morbidity or Mortality. by Ebong SJ, Goyert SM, Nemzek JA, Kim J, Bolgos GL, Remick DG.; 2001 Apr; http://www.pubmedcentral.gov/articlerender.fcgi?tool=pmcentrez&artid=98135
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Dantrolene Ameliorates the Metabolic Hallmarks of Sepsis in Rats and Improves Survival in a Mouse Model of Endotoxemia. by Hotchkiss RS, Karl IE.; 1994 Apr 12; http://www.pubmedcentral.gov/articlerender.fcgi?tool=pmcentrez&rendertype=abstr act&artid=43510
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DF-2 septicemia following whirlpool spa immersion. by Holmes RL, Kozinn WP.; 1986 Mar; http://www.pubmedcentral.gov/picrender.fcgi?tool=pmcentrez&action=stream&blobt ype=pdf&artid=268709
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Differential Host Inflammatory Responses to Viable Versus Antibiotic-Killed Bacteria in Experimental Microbial Sepsis. by Silverstein R, Wood JG, Xue Q, Norimatsu M, Horn DL, Morrison DC.; 2000 Apr; http://www.pubmedcentral.gov/articlerender.fcgi?tool=pmcentrez&artid=97417
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Effect of Interleukin-10 on Gut-Derived Sepsis Caused by Pseudomonas aeruginosa in Mice. by Matsumoto T, Tateda K, Miyazaki S, Furuya N, Ohno A, Ishii Y, Hirakata Y, Yamaguchi K.; 1998 Nov; http://www.pubmedcentral.gov/articlerender.fcgi?tool=pmcentrez&artid=105955
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Effective Immunomodulatory Treatment of Escherichia coli Experimental Sepsis with Thalidomide. by Giamarellos-Bourboulis EJ, Poulaki H, Kostomitsopoulos N, Dontas I, Perrea D, Karayannacos PE, Giamarellou H.; 2003 Aug; http://www.pubmedcentral.gov/articlerender.fcgi?tool=pmcentrez&artid=166096
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Effects of Specific Antibodies against Streptococcus pneumoniae on Pharmacodynamic Parameters of [beta]-Lactams in a Mouse Sepsis Model. by Casal J, Aguilar L, Jado I, Yuste J, Gimenez MJ, Prieto J, Fenoll A.; 2002 May; http://www.pubmedcentral.gov/articlerender.fcgi?tool=pmcentrez&artid=127147
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Elevated Levels of Lipopolysaccharide-Binding Protein and Soluble CD14 in Plasma in Neonatal Early-Onset Sepsis. by Berner R, Furll B, Stelter F, Drose J, Muller HP, Schutt C.; 2002 Mar; http://www.pubmedcentral.gov/articlerender.fcgi?tool=pmcentrez&artid=119928
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Ethyl pyruvate prevents lethality in mice with established lethal sepsis and systemic inflammation. by Ulloa L, Ochani M, Yang H, Tanovic M, Halperin D, Yang R, Czura CJ, Fink MP, Tracey KJ.; 2002 Sep 17; http://www.pubmedcentral.gov/articlerender.fcgi?tool=pmcentrez&artid=129448
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Expression of the C5a receptor (CD88) on granulocytes and monocytes in patients with severe sepsis. by Furebring M, Hakansson LD, Venge P, Nilsson B, Sjolin J.; 2002; http://www.pubmedcentral.gov/articlerender.fcgi?tool=pmcentrez&artid=125318
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Fatal sepsis caused by Corynebacterium amycolatum in a premature infant. by Berner R, Pelz K, Wilhelm C, Funke A, Leititis JU, Brandis M.; 1997 Apr; http://www.pubmedcentral.gov/articlerender.fcgi?tool=pmcentrez&rendertype=abstr act&artid=229725
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Genetic Characterization of Resistance to Extended-Spectrum [beta]-Lactams in Klebsiella oxytoca Isolates Recovered from Patients with Septicemia at Hospitals in the Stockholm Area. by Wu SW, Dornbusch K, Kronvall G.; 1999 May; http://www.pubmedcentral.gov/articlerender.fcgi?tool=pmcentrez&artid=89264
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Gram-negative sepsis: a dilemma of modern medicine. by Bone RC.; 1993 Jan; http://www.pubmedcentral.gov/picrender.fcgi?tool=pmcentrez&action=stream&blobt ype=pdf&artid=358266
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Heat stress is associated with decreased lactic acidemia in rat sepsis. by Deshpande GG, Heidemann SM, Sarnaik AP.; 2000; http://www.pubmedcentral.gov/articlerender.fcgi?tool=pmcentrez&artid=29035
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Hemofiltration in sepsis: where do we go from here? by Kellum JA, Bellomo R.; 2000; http://www.pubmedcentral.gov/articlerender.fcgi?tool=pmcentrez&artid=137329
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Highly sensitive immunoassay for direct diagnosis of viral hemorrhagic septicemia which uses antinucleocapsid monoclonal antibodies. by Mourton C, Romestand B, de Kinkelin P, Jeffroy J, Le Gouvello R, Pau B.; 1992 Sep; http://www.pubmedcentral.gov/picrender.fcgi?tool=pmcentrez&action=stream&blobt ype=pdf&artid=265503
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Hot new therapy for sepsis and the acute respiratory distress syndrome. by Slutsky AS.; 2002 Sep 15; http://www.pubmedcentral.gov/articlerender.fcgi?tool=pmcentrez&artid=151137
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Immunomodulating effect of fosfomycin on gut-derived sepsis caused by Pseudomonas aeruginosa in mice. by Matsumoto T, Tateda K, Miyazaki S, Furuya N, Ohno A, Ishii Y, Hirakata Y, Yamaguchi K.; 1997 Feb; http://www.pubmedcentral.gov/articlerender.fcgi?tool=pmcentrez&rendertype=abstr act&artid=163706
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Immunopathologic Alterations in Murine Models of Sepsis of Increasing Severity. by Ebong S, Call D, Nemzek J, Bolgos G, Newcomb D, Remick D.; 1999 Dec; http://www.pubmedcentral.gov/articlerender.fcgi?tool=pmcentrez&artid=97073
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Inactivation of catecholamines by superoxide gives new insights on the pathogenesis of septic shock. by Macarthur H, Westfall TC, Riley DP, Misko TP, Salvemini D.; 2000 Aug 15; http://www.pubmedcentral.gov/articlerender.fcgi?tool=pmcentrez&artid=16937
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Increased C5a receptor expression in sepsis. by Riedemann NC, Guo RF, Neff TA, Laudes IJ, Keller KA, Sarma VJ, Markiewski MM, Mastellos D, Strey CW, Pierson CL, Lambris JD, Zetoune FS, Ward PA.; 2002 Jul 1; http://www.pubmedcentral.gov/articlerender.fcgi?tool=pmcentrez&artid=151030
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Infection of Neonatal Mice with Sindbis Virus Results in a Systemic Inflammatory Response Syndrome. by Klimstra WB, Ryman KD, Bernard KA, Nguyen KB, Biron CA, Johnston RE.; 1999 Dec; http://www.pubmedcentral.gov/articlerender.fcgi?tool=pmcentrez&artid=113094
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Inhibition of Leukocyte Rolling by Nitric Oxide during Sepsis Leads to Reduced Migration of Active Microbicidal Neutrophils. by Benjamim CF, Silva JS, Fortes ZB, Oliveira MA, Ferreira SH, Cunha FQ.; 2002 Jul; http://www.pubmedcentral.gov/articlerender.fcgi?tool=pmcentrez&artid=128083
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Interleukin-10 Controls the Onset of Irreversible Septic Shock. by Latifi SQ, O'Riordan MA, Levine AD.; 2002 Aug; http://www.pubmedcentral.gov/articlerender.fcgi?tool=pmcentrez&artid=128185
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Interleukin-8 in sepsis: relation to shock and inflammatory mediators. by Hack CE, Hart M, van Schijndel RJ, Eerenberg AJ, Nuijens JH, Thijs LG, Aarden LA.; 1992 Jul; http://www.pubmedcentral.gov/picrender.fcgi?tool=pmcentrez&action=stream&blobt ype=pdf&artid=257242
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Levels of Inhibitors of Tumor Necrosis Factor Alpha and Interleukin 1[beta] in Urine and Sera of Patients with Urosepsis. by Olszyna DP, Prins JM, Buis B, van Deventer SJ, Speelman P, van der Poll T.; 1998 Aug; http://www.pubmedcentral.gov/articlerender.fcgi?tool=pmcentrez&artid=108383
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Lipopolyamines: Novel Antiendotoxin Compounds That Reduce Mortality in Experimental Sepsis Caused by Gram-Negative Bacteria. by David SA, Silverstein R, Amura CR, Kielian T, Morrison DC.; 1999 Apr; http://www.pubmedcentral.gov/articlerender.fcgi?tool=pmcentrez&artid=89225
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Mass transfer, clearance and plasma concentration of procalcitonin during continuous venovenous hemofiltration in patients with septic shock and acute oliguric renal failure. by Level C, Chauveau P, Guisset O, Cazin MC, Lasseur C, Gabinsky C, Winnock S, Montaudon D, Bedry R, Nouts C, Pillet O, Benissan GG, Favarel-Guarrigues JC, Castaing Y.; 2003; http://www.pubmedcentral.gov/articlerender.fcgi?tool=pmcentrez&artid=374372
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Mast cell dipeptidyl peptidase I mediates survival from sepsis. by Mallen --St. Clair J, Pham CT, Villalta SA, Caughey GH, Wolters PJ.; 2004 Feb 15; http://www.pubmedcentral.gov/articlerender.fcgi?tool=pmcentrez&artid=338261
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Mechanisms Involved in the Pathogenesis of Sepsis Are Not Necessarily Reflected by In Vitro Cell Activation Studies. by Amura CR, Silverstein R, Morrison DC.; 1998 Nov; http://www.pubmedcentral.gov/articlerender.fcgi?tool=pmcentrez&artid=108672
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Modulation of Release of Proinflammatory Bacterial Compounds by Antibacterials: Potential Impact on Course of Inflammation and Outcome in Sepsis and Meningitis. by Nau R, Eiffert H.; 2002 Jan; http://www.pubmedcentral.gov/articlerender.fcgi?tool=pmcentrez&artid=118062
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Molecular Cloning and Characterization of the afa-7 and afa-8 Gene Clusters Encoding Afimbrial Adhesins in Escherichia coli Strains Associated with Diarrhea or Septicemia in Calves. by Lalioui L, Jouve M, Gounon P, Le Bouguenec C.; 1999 Oct; http://www.pubmedcentral.gov/articlerender.fcgi?tool=pmcentrez&artid=96852
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Neutrophils employ the myeloperoxidase system to generate antimicrobial brominating and chlorinating oxidants during sepsis. by Gaut JP, Yeh GC, Tran HD, Byun J, Henderson JP, Richter GM, Brennan ML, Lusis AJ, Belaaouaj A, Hotchkiss RS, Heinecke JW.; 2001 Oct 9; http://www.pubmedcentral.gov/articlerender.fcgi?tool=pmcentrez&artid=59821
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Nosocomial septicemia caused by Serratia plymuthica. by Domingo D, Limia A, Alarcon T, Sanz JC, Del Rey MC, Lopez-Brea M.; 1994 Feb; http://www.pubmedcentral.gov/picrender.fcgi?tool=pmcentrez&action=stream&blobt ype=pdf&artid=263083
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papG Alleles among Escherichia coli Strains Causing Urosepsis: Associations with Other Bacterial Characteristics and Host Compromise. by Johnson JR.; 1998 Sep; http://www.pubmedcentral.gov/articlerender.fcgi?tool=pmcentrez&artid=108561
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Pasteurella multocida septicemia and subsequent Pasteurella dagmatis septicemia in a diabetic patient. by Fajfar-Whetstone CJ, Coleman L, Biggs DR, Fox BC.; 1995 Jan; http://www.pubmedcentral.gov/articlerender.fcgi?tool=pmcentrez&rendertype=abstr act&artid=227908
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Pathogenicity of an Escherichia coli O115:K"V165" mutant negative for F165(1) fimbriae in septicemia of gnotobiotic pigs. by Ngeleka M, Jacques M, Martineau-Doize B, Daigle F, Harel J, Fairbrother JM.; 1993 Mar; http://www.pubmedcentral.gov/picrender.fcgi?tool=pmcentrez&action=stream&blobt ype=pdf&artid=302809
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Pattern of cytokines and pharmacomodulation in sepsis induced by cecal ligation and puncture compared with that induced by endotoxin. by Villa P, Sartor G, Angelini M, Sironi M, Conni M, Gnocchi P, Isetta AM, Grau G, Buurman W, van Tits LJ.; 1995 Sep; http://www.pubmedcentral.gov/articlerender.fcgi?tool=pmcentrez&rendertype=abstr act&artid=170198
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Phylogenetic Analysis and Prevalence of Urosepsis Strains of Escherichia coli Bearing Pathogenicity Island-Like Domains. by Bingen-Bidois M, Clermont O, Bonacorsi S, Terki M, Brahimi N, Loukil C, Barraud D, Bingen E.; 2002 Jun; http://www.pubmedcentral.gov/articlerender.fcgi?tool=pmcentrez&artid=128014
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Physiological-dose steroid therapy in sepsis [ISRCTN36253388]. by Yildiz O, Doganay M, Aygen B, Guven M, Kelestimur F, Tutus A.; 2002; http://www.pubmedcentral.gov/articlerender.fcgi?tool=pmcentrez&artid=125315
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Potential Therapeutic Role of Cationic Peptides in Three Experimental Models of Septic Shock. by Giacometti A, Cirioni O, Ghiselli R, Mocchegiani F, Del Prete MS, Viticchi C, Kamysz W, Lempicka E, Saba V, Scalise G.; 2002 Jul; http://www.pubmedcentral.gov/articlerender.fcgi?tool=pmcentrez&artid=127283
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Predictive Value of Nuclear Factor [kappa]B Activity and Plasma Cytokine Levels in Patients with Sepsis. by Arnalich F, Garcia-Palomero E, Lopez J, Jimenez M, Madero R, Renart J, Vazquez JJ, Montiel C.; 2000 Apr; http://www.pubmedcentral.gov/articlerender.fcgi?tool=pmcentrez&artid=97370
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Prevalence of Molecular Types and mecA Gene Carriage of Coagulase-Negative Staphylococci in a Neonatal Intensive Care Unit: Relation to Nosocomial Septicemia. by Krediet TG, Jones ME, Janssen K, Gerards LJ, Fleer A.; 2001 Sep; http://www.pubmedcentral.gov/articlerender.fcgi?tool=pmcentrez&artid=88351
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Prevention of lymphocyte cell death in sepsis improves survival in mice. by Hotchkiss RS, Tinsley KW, Swanson PE, Chang KC, Cobb JP, Buchman TG, Korsmeyer SJ, Karl IE.; 1999 Dec 7; http://www.pubmedcentral.gov/articlerender.fcgi?tool=pmcentrez&artid=24472
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Primary Shewanella alga Septicemia in a Patient on Hemodialysis. by Iwata M, Tateda K, Matsumoto T, Furuya N, Mizuiri S, Yamaguchi K.; 1999 Jun; http://www.pubmedcentral.gov/articlerender.fcgi?tool=pmcentrez&artid=85050
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Pro/con clinical debate: Is high-volume hemofiltration beneficial in the treatment of septic shock? by Reiter K, Bellomo R, Ronco C, Kellum JA.; 2002; http://www.pubmedcentral.gov/articlerender.fcgi?tool=pmcentrez&artid=137392
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Pulmonary and Hepatic Gene Expression following Cecal Ligation and Puncture: Monophosphoryl Lipid A Prophylaxis Attenuates Sepsis-Induced Cytokine and Chemokine Expression and Neutrophil Infiltration. by Salkowski CA, Detore G, Franks A, Falk MC, Vogel SN.; 1998 Aug; http://www.pubmedcentral.gov/articlerender.fcgi?tool=pmcentrez&artid=108388
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Quality of life effects of antithrombin III in sepsis survivors: results from the KyberSept trial [ISRCTN22931023]. by Rublee D, Opal SM, Schramm W, Keinecke HO, Knaub S.; 2002; http://www.pubmedcentral.gov/articlerender.fcgi?tool=pmcentrez&artid=125322
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Quantitative aspects of septicemia. by Yagupsky P, Nolte FS.; 1990 Jul; http://www.pubmedcentral.gov/picrender.fcgi?tool=pmcentrez&action=stream&blobt ype=pdf&artid=358159
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Reappraisal with meta-analysis of bacteremia, endotoxemia, and mortality in gramnegative sepsis. by Hurley JC.; 1995 May; http://www.pubmedcentral.gov/articlerender.fcgi?tool=pmcentrez&rendertype=abstr act&artid=228145
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Risk factors for early onset neonatal group B streptococcal sepsis: case-control study. by Oddie S, Embleton ND.; 2002 Aug 10; http://www.pubmedcentral.gov/articlerender.fcgi?tool=pmcentrez&artid=117770
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Role of endotoxemia in cardiovascular dysfunction and lethality: virulent and nonvirulent Escherichia coli challenges in a canine model of septic shock. by Hoffman WD, Danner RL, Quezado ZM, Banks SM, Elin RJ, Hosseini JM, Natanson C.; 1996 Feb; http://www.pubmedcentral.gov/articlerender.fcgi?tool=pmcentrez&rendertype=abstr act&artid=173778
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Role of IL-12 in Staphylococcus aureus-triggered arthritis and sepsis. by Hultgren OH, Stenson M, Tarkowski A.; 2001; http://www.pubmedcentral.gov/articlerender.fcgi?tool=pmcentrez&artid=17823
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Safety assessment of drotrecogin alfa (activated) in the treatment of adult patients with severe sepsis. by Bernard GR, Macias WL, Joyce DE, Williams MD, Bailey J, Vincent JL.; 2003; http://www.pubmedcentral.gov/articlerender.fcgi?tool=pmcentrez&artid=270661
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Sepsis, Multiple Organ Failure, and Death Due to Pandoraea pnomenusa Infection after Lung Transplantation. by Stryjewski ME, LiPuma JJ, Messier, Jr. RH, Reller LB, Alexander BD.; 2003 May; http://www.pubmedcentral.gov/articlerender.fcgi?tool=pmcentrez&artid=154699
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Septic Shock Due to Helicobacter fennelliae in a Non-Human Immunodeficiency Virus-Infected Heterosexual Patient. by Hsueh PR, Teng LJ, Hung CC, Chen YC, Yang PC, Ho SW, Luh KT.; 1999 Jun; http://www.pubmedcentral.gov/articlerender.fcgi?tool=pmcentrez&artid=85042
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Septicemia caused by the gram-negative bacterium CDC IV c-2 in an immunocompromised human. by Dan M, Berger SA, Aderka D, Levo Y.; 1986 Apr; http://www.pubmedcentral.gov/picrender.fcgi?tool=pmcentrez&action=stream&blobt ype=pdf&artid=362844
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Septicemia Due to Acinetobacter junii. by Linde HJ, Hahn J, Holler E, Reischl U, Lehn N.; 2002 Jul; http://www.pubmedcentral.gov/articlerender.fcgi?tool=pmcentrez&artid=120562
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Septicemia Due to Pasteurella pneumotropica: 16S rRNA Sequencing for Diagnosis Confirmation. by Frebourg NB, Berthelot G, Hocq R, Chibani A, Lemeland JF.; 2002 Feb; http://www.pubmedcentral.gov/articlerender.fcgi?tool=pmcentrez&artid=153411
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Septicemia in Neutropenic Patients Infected with Clostridium tertium Resistant to Cefepime and Other Expanded-Spectrum Cephalosporins. by Steyaert S, Peleman R, Vaneechoutte M, De Baere T, Claeys G, Verschraegen G.; 1999 Nov; http://www.pubmedcentral.gov/articlerender.fcgi?tool=pmcentrez&artid=85761
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Serial Granulocyte Transfusions as a Treatment for Sepsis Due to MultidrugResistant Pseudomonas aeruginosa in a Neutropenic Patient. by Lin YW, Adachi S, Watanabe KI, Umeda K, Nakahata T.; 2003 Oct; http://www.pubmedcentral.gov/articlerender.fcgi?tool=pmcentrez&artid=254381
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Severe sepsis: variation in resource and therapeutic modality use among academic centers. by Yu DT, Black E, Sands KE, Schwartz JS, Hibberd PL, Graman PS, Lanken PN, Kahn KL, Snydman DR, Parsonnet J, Moore R, Platt R, Bates DW.; 2003; http://www.pubmedcentral.gov/articlerender.fcgi?tool=pmcentrez&artid=270675
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Severe Serratia liquefaciens Sepsis following Vitamin C Infusion Treatment by a Naturopathic Practitioner. by Engelhart S, Saborowski F, Krakau M, ScherholzSchlosser G, Heyer I, Exner M.; 2003 Aug; http://www.pubmedcentral.gov/articlerender.fcgi?tool=pmcentrez&artid=179848
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Simultaneous Detection of Neisseria meningitidis, Haemophilus influenzae, and Streptococcus pneumoniae in Suspected Cases of Meningitis and Septicemia Using Real-Time PCR. by Corless CE, Guiver M, Borrow R, Edwards-Jones V, Fox AJ, Kaczmarski EB.; 2001 Apr; http://www.pubmedcentral.gov/articlerender.fcgi?tool=pmcentrez&artid=87969
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STAT4 Is Required for Antibacterial Defense but Enhances Mortality during Polymicrobial Sepsis. by Godshall CJ, Lentsch AB, Peyton JC, Scott MJ, Cheadle WG.; 2001 Nov; http://www.pubmedcentral.gov/articlerender.fcgi?tool=pmcentrez&artid=96223
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Susceptibility testing of macrolide antibiotics against Haemophilus influenzae and correlation of in vitro results with in vivo efficacy in a mouse septicemia model. by Fernandes PB, Hardy D, Bailer R, McDonald E, Pintar J, Ramer N, Swanson R, Gade E.; 1987 Aug; http://www.pubmedcentral.gov/picrender.fcgi?tool=pmcentrez&action=stream&blobt ype=pdf&artid=174912
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Synergistic effect of a recombinant N-terminal fragment of bactericidal/permeabilityincreasing protein and cefamandole in treatment of rabbit gram-negative sepsis. by Lin Y, Leach WJ, Ammons WS.; 1996 Jan; http://www.pubmedcentral.gov/articlerender.fcgi?tool=pmcentrez&rendertype=abstr act&artid=163058
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Targeted adenovirus-induced expression of IL-10 decreases thymic apoptosis and improves survival in murine sepsis. by Oberholzer C, Oberholzer A, Bahjat FR, Minter RM, Tannahill CL, Abouhamze A, LaFace D, Hutchins B, Clare-Salzler MJ, Moldawer LL.; 2001 Sep 25; http://www.pubmedcentral.gov/articlerender.fcgi?tool=pmcentrez&artid=58759
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The effects of IgM-enriched immunoglobulin preparations in patients with severe sepsis [ISRCTN28863830]. by Tugrul S, Ozcan PE, Akinci O, Seyhun Y, Cagatay A, Cakar N, Esen F.; 2002; http://www.pubmedcentral.gov/articlerender.fcgi?tool=pmcentrez&artid=125317
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The enigma of sepsis. by Riedemann NC, Guo RF, Ward PA.; 2003 Aug 15; http://www.pubmedcentral.gov/articlerender.fcgi?tool=pmcentrez&artid=171398
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The level of lipopolysaccharide-binding protein is significantly increased in plasma in patients with the systemic inflammatory response syndrome. by Myc A, Buck J, Gonin J, Reynolds B, Hammerling U, Emanuel D.; 1997 Mar; http://www.pubmedcentral.gov/articlerender.fcgi?tool=pmcentrez&rendertype=abstr act&artid=170487
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The role of staphylococcal polysaccharide microcapsule expression in septicemia and septic arthritis. by Nilsson IM, Lee JC, Bremell T, Ryden C, Tarkowski A.; 1997 Oct; http://www.pubmedcentral.gov/articlerender.fcgi?tool=pmcentrez&rendertype=abstr act&artid=175605
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The Surviving Sepsis Campaign: raising awareness to reduce mortality. by Slade E, Tamber PS, Vincent JL.; 2003; http://www.pubmedcentral.gov/articlerender.fcgi?tool=pmcentrez&artid=154124
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Tissue distribution of migration inhibitory factor and inducible nitric oxide synthase in falciparum malaria and sepsis in African children. by Clark IA, Awburn MM, Whitten RO, Harper CG, Liomba NG, Molyneux ME, Taylor TE.; 2003; http://www.pubmedcentral.gov/articlerender.fcgi?tool=pmcentrez&artid=154094
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Transfusion-associated bacterial sepsis. by Wagner SJ, Friedman LI, Dodd RY.; 1994 Jul; http://www.pubmedcentral.gov/picrender.fcgi?tool=pmcentrez&action=stream&blobt ype=pdf&artid=358326
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Treatment of Klebsiella pneumoniae septicemia in normal and leukopenic mice by liposome-encapsulated muramyl tripeptide phosphatidylethanolamide. by Melissen PM, van Vianen W, Bakker-Woudenberg IA.; 1994 Jan; http://www.pubmedcentral.gov/picrender.fcgi?tool=pmcentrez&action=stream&blobt ype=pdf&artid=284413
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Use of Lytic Bacteriophage for Control of Experimental Escherichia coli Septicemia and Meningitis in Chickens and Calves. by Barrow P, Lovell M, Berchieri A Jr.; 1998 May; http://www.pubmedcentral.gov/articlerender.fcgi?tool=pmcentrez&artid=104512
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Usefulness of gram staining of blood collected from total parenteral nutrition catheter for rapid diagnosis of catheter-related sepsis. by Moonens F, el Alami S, Van Gossum A, Struelens MJ, Serruys E.; 1994 Jun; http://www.pubmedcentral.gov/picrender.fcgi?tool=pmcentrez&action=stream&blobt ype=pdf&artid=264041
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Usefulness of procalcitonin for diagnosis of sepsis in the intensive care unit. by BalcI C, Sungurtekin H, Gurses E, Sungurtekin U, Kaptanoglu B.; 2003; http://www.pubmedcentral.gov/articlerender.fcgi?tool=pmcentrez&artid=154110
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Utilization of 16S Ribosomal DNA Sequencing for Diagnosis of Septicemia Due to Neisseria elongata subsp. glycolytica in a Neutropenic Patient. by Hombrouck-Alet C, Poilane I, Janoir-Jouveshomme C, Fain O, Cruaud P, Thomas M, Collignon A.; 2003 Jul; http://www.pubmedcentral.gov/articlerender.fcgi?tool=pmcentrez&artid=165362
The National Library of Medicine: PubMed One of the quickest and most comprehensive ways to find academic studies in both English and other languages is to use PubMed, maintained by the National Library of Medicine.6 The advantage of PubMed over previously mentioned sources is that it covers a greater number of domestic and foreign references. It is also free to use. If the publisher has a Web site that offers full text of its journals, PubMed will provide links to that site, as well as to sites offering other related data. User registration, a subscription fee, or some other type of fee may be required to access the full text of articles in some journals. To generate your own bibliography of studies dealing with sepsis, simply go to the PubMed Web site at http://www.ncbi.nlm.nih.gov/pubmed. Type “sepsis” (or synonyms) into the search box, and click “Go.” The following is the type of output you can expect from PubMed for sepsis (hyperlinks lead to article summaries): •
A case of Pasteurella haemolytica sepsis in a patient with mitral valve disease who developed a splenic abscess. Author(s): Takeda S, Arashima Y, Kato K, Ogawa M, Kono K, Watanabe K, Saito T. Source: Scandinavian Journal of Infectious Diseases. 2003; 35(10): 764-5. http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=A bstract&list_uids=14606620
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A definite role for treatment with activated protein C in sepsis? Standard use is premature. Author(s): Girbes AR, Polderman KH. Source: Journal of Thrombosis and Haemostasis : Jth. 2003 December; 1(12): 2469-71. http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=A bstract&list_uids=14675079
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PubMed was developed by the National Center for Biotechnology Information (NCBI) at the National Library of Medicine (NLM) at the National Institutes of Health (NIH). The PubMed database was developed in conjunction with publishers of biomedical literature as a search tool for accessing literature citations and linking to full-text journal articles at Web sites of participating publishers. Publishers that participate in PubMed supply NLM with their citations electronically prior to or at the time of publication.
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A definite role for treatment with activated protein C in sepsis? Yes. Author(s): Wiel E, Vallet B. Source: Journal of Thrombosis and Haemostasis : Jth. 2003 December; 1(12): 2466-8. http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=A bstract&list_uids=14675078
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A new weapon against severe sepsis related to necrotizing fasciitis. Author(s): Purnell D, Hazlett T, Alexander SL. Source: Dimensions of Critical Care Nursing : Dccn. 2004 January-February; 23(1): 18-23. http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=A bstract&list_uids=14734896
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A patient with E. coli-induced pyelonephritis and sepsis who transiently exhibited symptoms associated with primary biliary cirrhosis. Author(s): Ohno N, Ota Y, Hatakeyama S, Yanagimoto S, Morisawa Y, Tsukada K, Koike K, Kimura S. Source: Intern Med. 2003 November; 42(11): 1144-8. http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=A bstract&list_uids=14686759
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A readers' guide to the interpretation of diagnostic test properties: clinical example of sepsis. Author(s): Fischer JE, Bachmann LM, Jaeschke R. Source: Intensive Care Medicine. 2003 July; 29(7): 1043-51. Epub 2003 May 07. Review. http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=A bstract&list_uids=12734652
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Abdominal sepsis and communication breakdown. Author(s): McCoy R. Source: Aust Fam Physician. 2003 May; 32(5): 345-6. No Abstract Available. http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=A bstract&list_uids=12772367
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Acute acalculous cholecystitis associated with systemic sepsis and visceral arterial hypoperfusion: a case series and review of pathophysiology. Author(s): McChesney JA, Northup PG, Bickston SJ. Source: Digestive Diseases and Sciences. 2003 October; 48(10): 1960-7. http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=A bstract&list_uids=14627341
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Adequacy of early empiric antibiotic treatment and survival in severe sepsis: experience from the MONARCS trial. Author(s): MacArthur RD, Miller M, Albertson T, Panacek E, Johnson D, Teoh L, Barchuk W. Source: Clinical Infectious Diseases : an Official Publication of the Infectious Diseases Society of America. 2004 January 15; 38(2): 284-8. Epub 2003 December 22. http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=A bstract&list_uids=14699463
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Administration of the nitric oxide synthase inhibitor NG-methyl-L-arginine hydrochloride (546C88) by intravenous infusion for up to 72 hours can promote the resolution of shock in patients with severe sepsis: results of a randomized, doubleblind, placebo-controlled multicenter study (study no. 144-002). Author(s): Bakker J, Grover R, McLuckie A, Holzapfel L, Andersson J, Lodato R, Watson D, Grossman S, Donaldson J, Takala J; Glaxo Wellcome International Septic Shock Study Group. Source: Critical Care Medicine. 2004 January; 32(1): 1-12. http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=A bstract&list_uids=14707554
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Adrenocortical hormones in survivors and nonsurvivors of severe sepsis: diverse time course of dehydroepiandrosterone, dehydroepiandrosterone-sulfate, and cortisol. Author(s): Marx C, Petros S, Bornstein SR, Weise M, Wendt M, Menschikowski M, Engelmann L, Hoffken G. Source: Critical Care Medicine. 2003 May; 31(5): 1382-8. http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=A bstract&list_uids=12771606
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Aminoglycoside and vancomycin serum concentration monitoring and mortality due to neonatal sepsis in Saudi Arabia. Author(s): El Desoky ES, Sheikh AA, Al Hammadi AY. Source: Journal of Clinical Pharmacy and Therapeutics. 2003 December; 28(6): 479-83. http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=A bstract&list_uids=14651671
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Analysis of sepsis in allogeneic bone marrow transplant recipients: a single-center study. Author(s): Mitsui H, Karasuno T, Santo T, Fukushima K, Matsunaga H, Nakamura H, Hiraoka A. Source: Journal of Infection and Chemotherapy : Official Journal of the Japan Society of Chemotherapy. 2003 September; 9(3): 238-42. http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=A bstract&list_uids=14513392
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Analysis of the efficacy of urine culture as part of sepsis evaluation in the premature infant. Author(s): Tamim MM, Alesseh H, Aziz H. Source: The Pediatric Infectious Disease Journal. 2003 September; 22(9): 805-8. http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=A bstract&list_uids=14506372
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Antibody to enterobacterial common antigen and gram-negative sepsis. Author(s): Bayardelle P, Albertson TE, MacArthur RD. Source: Critical Care Medicine. 2004 February; 32(2): 617-8. http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=A bstract&list_uids=14758202
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Anti-complement strategies in experimental sepsis. Author(s): Ward PA, Riedemann NC, Guo RF, Huber-Lang M, Sarma JV, Zetoune FS. Source: Scandinavian Journal of Infectious Diseases. 2003; 35(9): 601-3. http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=A bstract&list_uids=14620141
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Antioxidant therapy for severe sepsis: promise and perspective. Author(s): Pinsky MR. Source: Critical Care Medicine. 2003 November; 31(11): 2697-8. Review. http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=A bstract&list_uids=14605544
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Arterial hypertension and sepsis. Author(s): Nunes JP. Source: Rev Port Cardiol. 2003 November; 22(11): 1375-9. Review. http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=A bstract&list_uids=14768492
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Assessment of six mortality prediction models in patients admitted with severe sepsis and septic shock to the intensive care unit: a prospective cohort study. Author(s): Arabi Y, Al Shirawi N, Memish Z, Venkatesh S, Al-Shimemeri A. Source: Critical Care (London, England). 2003 October; 7(5): R116-22. Epub 2003 August 28. http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=A bstract&list_uids=12974979
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Association of mannose-binding lectin polymorphisms with sepsis and fatal outcome, in patients with systemic inflammatory response syndrome. Author(s): Garred P, J Strom J, Quist L, Taaning E, Madsen HO. Source: The Journal of Infectious Diseases. 2003 November 1; 188(9): 1394-403. Epub 2003 October 15. http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=A bstract&list_uids=14593599
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Bacillus Calmette-Guerin sepsis: shift of an intended local toward a detrimental systemic cytotoxic immune response. Author(s): Zeerleder S, Hack CE, Caliezi C, Hebeisen R, Wuillemin WA. Source: Blood. 2001 August 1; 98(3): 890-1. http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=A bstract&list_uids=11468141
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Bacteremia and sepsis due to Prevotella oris from dentoalveolar abscesses. Author(s): Bein T, Brem J, Schusselbauer T. Source: Intensive Care Medicine. 2003 May; 29(5): 856. Epub 2003 March 28. http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=A bstract&list_uids=12664220
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Bacteremic sepsis in intensive care: temporal trends in incidence, organ dysfunction, and prognosis. Author(s): Hugonnet S, Harbarth S, Ferriere K, Ricou B, Suter P, Pittet D. Source: Critical Care Medicine. 2003 February; 31(2): 390-4. http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=A bstract&list_uids=12576941
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Bacteria in blood smears: overwhelming sepsis or trivial contamination. Author(s): van der Meer W, Verwiel JM, Gidding CE, de Metz M, de Keijzer MH. Source: Acta Haematologica. 2002; 107(4): 220-3. http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=A bstract&list_uids=12053150
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Bad and good news in prevention and management of sepsis and MODS. Author(s): Baue AE. Source: Minerva Anestesiol. 2001 November; 67(11): 773-83. Review. No Abstract Available. http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=A bstract&list_uids=11753221
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Balance of inflammation in sepsis. Author(s): Pravinkumar E. Source: American Journal of Respiratory and Critical Care Medicine. 2004 March 1; 169(5): 655-6; Author Reply 656. http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=A bstract&list_uids=14982827
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Bench-to-bedside review: endothelial cell dysfunction in severe sepsis: a role in organ dysfunction? Author(s): Vallet B. Source: Critical Care (London, England). 2003 April; 7(2): 130-8. Epub 2003 January 06. Review. http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=A bstract&list_uids=12720559
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Bench-to-bedside review: functional relationships between coagulation and the innate immune response and their respective roles in the pathogenesis of sepsis. Author(s): Opal SM, Esmon CT. Source: Critical Care (London, England). 2003 February; 7(1): 23-38. Epub 2002 December 20. Review. http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=A bstract&list_uids=12617738
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Bench-to-bedside review: microvascular dysfunction in sepsis--hemodynamics, oxygen transport, and nitric oxide. Author(s): Bateman RM, Sharpe MD, Ellis CG. Source: Critical Care (London, England). 2003 October; 7(5): 359-73. Epub 2003 July 28. Review. http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=A bstract&list_uids=12974969
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Best evidence in anesthetic practice: prevention: recombinant human activated protein C reduces mortality in severe sepsis. Author(s): Butler R, Laufer B. Source: Canadian Journal of Anaesthesia = Journal Canadien D'anesthesie. 2002 February; 49(2): 207-9. http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=A bstract&list_uids=11823403
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Beta lactam monotherapy versus beta lactam-aminoglycoside combination therapy for sepsis in immunocompetent patients: systematic review and meta-analysis of randomised trials. Author(s): Paul M, Benuri-Silbiger I, Soares-Weiser K, Leibovici L. Source: Bmj (Clinical Research Ed.). 2004 March 20; 328(7441): 668. Epub 2004 Mar 02. Review. Erratum In: Bmj. 2004 April 10; 328(7444): 884. http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=A bstract&list_uids=14996699
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Beyond the complete blood cell count and C-reactive protein: a systematic review of modern diagnostic tests for neonatal sepsis. Author(s): Malik A, Hui CP, Pennie RA, Kirpalani H. Source: Archives of Pediatrics & Adolescent Medicine. 2003 June; 157(6): 511-6. Review. http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=A bstract&list_uids=12796229
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Bilateral endogenous endophthalmitis as a complication of late-onset sepsis in a premature infant. Author(s): Matasova K, Hudecova J, Zibolen M. Source: European Journal of Pediatrics. 2003 May; 162(5): 346-7. Epub 2003 February 27. http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=A bstract&list_uids=12692717
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Biliary sepsis in a patient on anti-TNFalpha therapy. Author(s): McNamara DA, Molony D, Mulsow J, O'Connell PR. Source: Ir J Med Sci. 2001 July-September; 170(3): 210-1. No Abstract Available. http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=A bstract&list_uids=12120982
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Biochemical markers of neonatal sepsis: value of procalcitonin in the emergency setting. Author(s): Guibourdenche J, Bedu A, Petzold L, Marchand M, Mariani-Kurdjian P, Marie-Fran, Hurtaud-Roux O, Aujard Y, Porquet D. Source: Annals of Clinical Biochemistry. 2002 March; 39(Pt 2): 130-5. http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=A bstract&list_uids=11928760
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Biomarkers of sepsis: is procalcitonin ready for prime time? Author(s): Pugin J. Source: Intensive Care Medicine. 2002 September; 28(9): 1203-4. http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=A bstract&list_uids=12400563
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Blood purification in sepsis: an idea whose time has come? Author(s): Kellum JA, Venkataraman R. Source: Critical Care Medicine. 2002 June; 30(6): 1387-8. http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=A bstract&list_uids=12072702
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Both gram positive and gram negative organisms complicate neutropaenic sepsis in children with leukemia. A unit audit study and recommendation. Author(s): Lee KY, Chan WS, Koay LS, Huang F, Tan LM, Yeoh AE. Source: Ann Acad Med Singapore. 2003 September; 32(5 Suppl): S37-8. No Abstract Available. http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=A bstract&list_uids=14968729
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Breast gangrene as a complication of puerperal sepsis. Author(s): Rege SA, Nunes Q, Rajput A, Dalvi AN. Source: Archives of Surgery (Chicago, Ill. : 1960). 2002 December; 137(12): 1441-2. http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=A bstract&list_uids=12470116
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Brevibacterium casei bacteremia and line sepsis in a patient with AIDS. Author(s): Janda WM, Tipirneni P, Novak RM. Source: The Journal of Infection. 2003 January; 46(1): 61-4. http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=A bstract&list_uids=12504612
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Carcinoma of the colon presenting as Streptococcus salivarius sepsis. Author(s): Afek S, Sperber AD, Almog Y. Source: Journal of Clinical Gastroenterology. 2004 January; 38(1): 86-7. http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=A bstract&list_uids=14679337
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Caspase-3 and -8 activation and cytokine removal with a novel cellulose triacetate super-permeable membrane in an in vitro sepsis model. Author(s): Bordoni V, Bolgan I, Brendolan A, Crepaldi C, Gastaldon F, D'intini V, Pilotto L, Inguaggiato P, Bonello M, Galloni E, Everard P, Bellomo R, Ronco C. Source: Int J Artif Organs. 2003 October; 26(10): 897-905. http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=A bstract&list_uids=14636005
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Cellular changes in association with defense mechanisms in intra-abdominal sepsis. Author(s): Pavlidis TE. Source: Minerva Chir. 2003 December; 58(6): 777-81. Review. http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=A bstract&list_uids=14663404
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Characterization of a novel rapidly growing Mycobacterium species associated with sepsis. Author(s): Hong T, Butler WR, Hollis F, Floyd MM, Toney SR, Tang YW, Steele C, Leggiadro RJ. Source: Journal of Clinical Microbiology. 2003 December; 41(12): 5650-3. http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=A bstract&list_uids=14662956
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Circulating adrenocorticotropic hormone (ACTH) and cortisol concentrations in normal, appropriate-for-gestational-age newborns versus those with sepsis and respiratory distress: Cortisol response to low-dose and standard-dose ACTH tests. Author(s): Soliman AT, Taman KH, Rizk MM, Nasr IS, Alrimawy H, Hamido MS. Source: Metabolism: Clinical and Experimental. 2004 February; 53(2): 209-14. http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=A bstract&list_uids=14767873
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Clinical prognostic markers in patients with severe sepsis: a prospective analysis of 139 consecutive cases. Author(s): Gogos CA, Lekkou A, Papageorgiou O, Siagris D, Skoutelis A, Bassaris HP. Source: The Journal of Infection. 2003 November; 47(4): 300-6. http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=A bstract&list_uids=14556754
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Clinical review: Drotrecogin alfa (activated) as adjunctive therapy for severe sepsis-practical aspects at the bedside and patient identification. Author(s): Laterre PF, Wittebole X. Source: Critical Care (London, England). 2003 December; 7(6): 445-50. Epub 2003 June 30. Review. http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=A bstract&list_uids=14624684
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Clinical review: extracorporeal blood purification in severe sepsis. Author(s): Venkataraman R, Subramanian S, Kellum JA. Source: Critical Care (London, England). 2003 April; 7(2): 139-45. Epub 2003 February 21. Review. http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=A bstract&list_uids=12720560
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Clinical trial design and outcomes in patients with severe sepsis. Author(s): Opal SM. Source: Shock (Augusta, Ga.). 2003 October; 20(4): 295-302. Review. http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=A bstract&list_uids=14501941
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Clostridium difficile colitis causing toxic megacolon, severe sepsis and multiple organ dysfunction syndrome. Author(s): Dobson G, Hickey C, Trinder J. Source: Intensive Care Medicine. 2003 June; 29(6): 1030. Epub 2003 May 07. Review. http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=A bstract&list_uids=12734650
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Coagulation cascade in sepsis: getting from bench to bedside? Author(s): Brown G. Source: Critical Care (London, England). 2003 April; 7(2): 117-8. Epub 2002 November 06. http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=A bstract&list_uids=12720555
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Coagulation disorders in patients with cirrhosis and severe sepsis. Author(s): Plessier A, Denninger MH, Consigny Y, Pessione F, Francoz C, Durand F, Francque S, Bezeaud A, Chauvelot-Moachon L, Lebrec D, Valla DC, Moreau R. Source: Liver International : Official Journal of the International Association for the Study of the Liver. 2003 December; 23(6): 440-8. http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=A bstract&list_uids=15002397
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Colistin for Klebsiella pneumoniae-associated sepsis. Author(s): Karabinis A, Paramythiotou E, Mylona-Petropoulou D, Kalogeromitros A, Katsarelis N, Kontopidou F, Poularas I, Malamou-Lada H. Source: Clinical Infectious Diseases : an Official Publication of the Infectious Diseases Society of America. 2004 January 1; 38(1): E7-9. Epub 2003 December 04. http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=A bstract&list_uids=14679468
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Comparison of surface swab cultures and quantitative tissue biopsy cultures to predict sepsis in burn patients: a prospective study. Author(s): Sjoberg T, Mzezewa S, Jonsson K, Robertson V, Salemark L. Source: The Journal of Burn Care & Rehabilitation. 2003 November-December; 24(6): 365-70. http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=A bstract&list_uids=14610420
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Contact activation by pathogenic bacteria: a virulence mechanism contributing to the pathophysiology of sepsis. Author(s): Herwald H, Morgelin M, Bjorck L. Source: Scandinavian Journal of Infectious Diseases. 2003; 35(9): 604-7. Review. http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=A bstract&list_uids=14620142
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Cortisol levels and mortality in severe sepsis. Author(s): Sam S, Corbridge TC, Mokhlesi B, Comellas AP, Molitch ME. Source: Clinical Endocrinology. 2004 January; 60(1): 29-35. http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=A bstract&list_uids=14678284
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Cost-effectiveness of recombinant human activated protein C and the influence of severity of illness in the treatment of patients with severe sepsis. Author(s): Fowler RA, Hill-Popper M, Stasinos J, Petrou C, Sanders GD, Garber AM. Source: Journal of Critical Care. 2003 September; 18(3): 181-91; Discussion 191-4. http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=A bstract&list_uids=14595571
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CpG DNA: trigger of sepsis, mediator of protection, or both? Author(s): Krieg AM. Source: Scandinavian Journal of Infectious Diseases. 2003; 35(9): 653-9. Review. http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=A bstract&list_uids=14620150
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Crazy-paving appearance associated with Streptococcus pneumoniae sepsis. Author(s): Ngo MH, Chen HT, Stark P. Source: Seminars in Respiratory Infections. 2003 September; 18(3): 220-2. http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=A bstract&list_uids=14505284
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Cytokine cascade in sepsis. Author(s): Cavaillon JM, Adib-Conquy M, Fitting C, Adrie C, Payen D. Source: Scandinavian Journal of Infectious Diseases. 2003; 35(9): 535-44. Review. http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=A bstract&list_uids=14620132
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Detection of circulating lipopolysaccharide-bound monocytes in children with gramnegative sepsis. Author(s): Takeshita S, Nakatani K, Tsujimoto H, Kawamura Y, Sekine I. Source: The Journal of Infectious Diseases. 2000 November; 182(5): 1549-52. Epub 2000 October 03. http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=A bstract&list_uids=11015235
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Development of an anti-core lipopolysaccharide vaccine for the prevention and treatment of sepsis. Author(s): Cross AS, Opal S, Cook P, Drabick J, Bhattacharjee A. Source: Vaccine. 2004 February 17; 22(7): 812-7. Review. http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=A bstract&list_uids=15040932
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Diagnosis of neonatal sepsis: a clinical and laboratory challenge. Author(s): Chiesa C, Panero A, Osborn JF, Simonetti AF, Pacifico L. Source: Clinical Chemistry. 2004 February; 50(2): 279-87. Review. http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=A bstract&list_uids=14752012
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Diagnostic value of cytokines and C-reactive protein in the first 24 hours of neonatal sepsis. Author(s): Laborada G, Rego M, Jain A, Guliano M, Stavola J, Ballabh P, Krauss AN, Auld PA, Nesin M. Source: American Journal of Perinatology. 2003 November; 20(8): 491-501. http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=A bstract&list_uids=14703598
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Dismutating the
