Free Radical and Antioxidant Protocols

M E T H O D S I N M O L E C U L A R B I O L O G Y "

John M. Walker, SeRies EDITOR

108. Free Radical and Antioxidant Protocols, edited by Donald Armstrong, 1998

107. Cytochrome P450 Protocols, edited by fan R. Phillips and Elizabeth A. Shephard, 1998

106. Receptor Binding Techniques, edited by Mary Keen, 1998 105. Phospholipid Signaling Protocols, edited by lan Bird, 1998 104. Mycoplasma Protocols, edited by Roger J. Miles and Robin

A. J. Nicholas, 1998 103. Pichia Protocols, edited by David R. Higgins and James

Cregg, 1998 102. Bioluminescence Methods and Protocols, edited by Robert

,4. LaRossa, 1998 101. Myobacteria Protocols, edited by Tanya Parish and Nell G.

Stoker, 1998 100. Nitric Oxide Protocols, edited by M. A. Titheradge, 1997 99. Human Cytoklnes and Cytokine Receptors, edited by Rena

Debuts, 1998 98. DNA Profiling Protocols, edited by James M. Thomson, 1997 97. Molecular Embryology: Methods and Protocols, edited by

Paul T. Sharpe and lvor Mason, 1998 96. Adhesion Proteins Protocols, edited by Etbabenu Dejana, 1997 95. DNA Topology and DNA Topolsomeruses: I1. Ensymology

and Topoisomerase Targetted Drugs, edited by Maryo,4nn Bjornsti, 1998

94. DNA Topology and DNA Topolsomerases: I. DN,4 Topol- ogy and Enzyme Purification, edited by Mary-,4nn Bjorusti, 1998

93. Protein Phosphatase Protocols, edited by John W Ludlow, 1997

92. PCR in Bioanalysis, edited by Stephen Meltzer, 1997 91. Flow Cytometry Protocols, edited by Mark J. Jaroszeski,

1998 90. Drag--DNA Interactions: Methods, Case Studies, and Pro-

tocols, edited by Keith R. Fox, 1997 89. Retiuold Protocols, edited by Christopher Redfern, 1997 88. Protein Targeting Protocols, edited by Roger ,4. Clegg, 1997 87. Combinatorial Peptide Library Protocols, edited by Shmuel

Cabilly, 1997 86. RNA Isolation and Characterization Protocols, edited by

Ralph Rapley, 1997 85. Differential Display Methods and Protocols, edited by Peng

Liang and Arthur B. Pardee, 1997 84. Transmembrane Signaling Protocols, edited by Dafna Bar-

Sagi, 1997 83. Receptor Signal Transduction Protocols, edited by R. ,4. J.

Challiss, 1997 82. Arabidopsis Protocols, edited by Jos~ M Martinez-Zapater

and Julia Salinas, 1998 81. Plant Virology Protocols, edited by Gary D. Faster, 1998 80. lmmunochemleol Protocols, SECOND zDtrloN, edited by John

Pound, 1998 79. Polyamine Protocols, edited by David M. L. Morgan, 1998 78. Antibacterial Peptide Protocols, edited by William M.

Sharer, I997

77. Protein Synthesis: Methods and Protocols, edited by Robin Martin, t998

76. Glycoanolysis Protocols, edited by Elizabeth F. Hounsel, 1998

75. Basic Cell Culture Protocols, edited by Jeffrey W Pollard and John M. Walker, 1997

74. Rlbozyme Protocols, edited by Philip C. Turner, 1997 73. Neuropeptide Protocols, edited by G. Brent Irvine and

Carvell H. Williams, 1997 72. Neurotransmitter Methods, edited by Richard C. Rayne, 1997 71. PRINS and In Situ PCR Protocols, edited by John R.

Gosden, 1997 70. Sequence Data Analysis Guidebook, edited by Simon R.

Swindell, 1997 69. eDNA Library Protocols, edited by lan G. Cowell and

Caroline A. Austin, 1997 68. Gene Isolation and Mapping Protocols, edited byJacqueline

Boultwood, 1997 67. PCR Cloning Protocols: From Molecular Cloning to Genetic

Engineering, edited by Bruce A. White, 1996 66. Epitope Mapping Protocols, edited by Glenn E. Morris, I996 65. PCR Sequencing Protocols, edited by Ralph Rapley, 1996 64. Protein Sequencing Protocols, edited by Bryan J. Smith, 1996 63. Recombinant Proteins: Detection and Isolation Protocols,

edited by Rocky S. Tuan, 1996 62. Recombinant Gone Expression Protocols, edited by Rocky

S. Tuan, 1996 61. Protein and Peptide Analysis by Mass Spectrometry,

edited by John R. Chapman, 1996 60. Protein NMR Protocols, edited by David G. Reid, 1996 59. Protein Purification Protocols, edited by Shaven Doonan,

1996 58. Basic DNA and RNA Protocols, edited by Adrian J.

Harwood, 1996 57. In Vitro Mutugenesis Protocols, edited by Michael K.

Trower, 1996 56. Crystallographic Methods and Protocols, edited by Chris-

tapher Jones, Barbara Mulloy, and Mark Sanderson, 1996 55. Plant Cdl Electroporatian and Electrofusion Protocols,

edited by Jac A. Nickolo~ 1995 54. YAC Protocols, edited by David Markie, 1995 53. Yeast Protocols: Methods in Cell and Molecular Biology,

edited by lvor H. Evans, 1996 52. Capillary Electropboresls: Principles, Instrumentation, and

Applications, edited by Kevin D. Altria, 1996 51. Antibody Engineering Protocols, edited by Sudhir Paul, 1995 50. Species Diagnostics Protocols: PCR and Other Nucleic Acid

Methods, edited by Justin P. Clapp, 1996 49. Plant Gone Transfer and Expression Protocols, edited by

Heddwyn Jones, 1995 48. Animal Cell Electroporutian and Electrofasion Protocols,

edited by Jac A. Nickoloff, 1995 47. Electroporation Protocols for Microorganisms, edited by

Jac A. Nickolo~ 1995

Free Radical and Antioxidant

Protocols

Edited by

Donald Armstrong State University of New York, Buffalo, NY

Humana Press ~(E Totowa, New Jersey

© 1998 Humana Press Inc. 999 Riverview Drive, Suite 208 Totowa, New Jersey 07512

All rights reserved. No part of this book may be reproduced, stored in a retrieval system, or transmitted in any form or by any means, electronic, mechanical, photocopying, microfilming, recording, or otherwise without written permission from the Publisher. Methods in Molecular Biology TM is a trademark of The Humana Press Inc.

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This publication is printed on acid-free paper. ( ~ ) ANSI Z39.48-1984 (American Standards Institute) Permanence of Paper for Printed Library Materials.

Cover illustration: Courtesy of Dr. Donald Armstrong.

Cover design by Patricia F. Cleary.

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Methods in molecular biology TM.

Free radical and antioxidant protocols / edited by Donald Armstrong. p. cm. -- (Methods in molecular biology'- ; v. 108) Includes bibliographic references and index. ISBN 0-89603-472-0 ( a l l paper) 1. Free radicals (Chemistry)--Pathophysiology--Laboratory manuals. 2. Antioxidants--

Physiological effect--Laboratory manuals. 3. Oxidation, Physiological--Laboratory manuals. 4. Stress (Physiology)--Research--Laboratory manuals. I. Armstrong, Donald 1933-. II. Series: Methods in Molecular Biology (Totowa, N J) ; 108.

RB 170.F68 1998 616.07--dc21 98-16172

CIP

Preface

From a historical perspective, it is interesting and appropriate that 50 years ago, Michaelis (1) first proposed that hydroxyl, peroxy radicals, and hydrogen peroxide could be expected as a result of normal oxidative metabo- lism. Two years later, the role of these reactive oxygen species (ROS) was demonstrated during lipid auto-oxidation, which was attributed to lipid peroxidation detected by the conjugated diene method (2) and by histochem- istry (3). In 1945, Dam and Granados (4) showed increased peroxidation of fats in vitamin E deficiency and in 1953, Tappel (5) demonstrated the protective effects of antioxidants (AOX) against oxygen toxicity. In 1954, Gerschman (6) described how the combination of hyperbaric oxygen and X-irradiation caused formation of oxygen radicals and showed a correlation with decreased survival time. Her observation that females were significantly less sensitive to this oxidation is the first indication of estrogen protection. In 1956, Harman (7) proposed his free radical theory of aging, and subsequently Yagi (8) devised a method to measure lipid hydroperoxides (LHP) in human serum, which inaugurated in vivo testing procedures.

From that era until the present, numerous investigators have made important contributions to our theoretical-conceptual understanding of these processes, as well as methodological development, the acute and chronic pathophysiological mechanisms, and therapeutic or nutritional intervention in these processes. Although carbon- and nitrogen-centered radicals play a role in the etiology and pathogenesis of disease, it is primarily ROS that initiate these events and subsequently, the level of aqueous- and lipid-phase antioxidants that influence the level of protection.

The rapid proliferation of biological publications on free radicals, LHP, and AOX in the literature since 1980 is shown in Fig. 1.

The idea for a book of techniques describing current state-of-the-art meth- odology and technology emerged from a 1993 international conference cover- ing the use of free radical assays in diagnostic medicine. Virtually every organ and body fluid have now been studied with respect to the normal concentra- tion of ROS and AOX. The development of experimental models has allowed their utilization in designing studies that examine basic mechanisms of human disease, the effect of AOX, and the assessment of risk factors using in vivo, ex vivo, in vitro, and in situ conditions. Summarized in Table 1, are the numbers

V

vi

6 0 0 0 .

4000" .u_

2000,

Preface

. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .

1966-75 1976-79 1980-85 1986-91 1992-96

year

Fig. 1.

of papers published on the various disorders in which oxidative stress has been implicated (from refs. 9 and 10; also see ref. 11 for other publications covering general and organ specific diseases, as well as the VERIS Research and Information Service, La Grange, IL 60525 which abstracts antioxidant-related papers).

The preparation of Free Radical and Antioxidant Protocols is an undertaking by 70 authors who present step-by-step information on a wide range of assays that can be utilized in the study of primary or secondary oxidative stress. Protocols in routine use are not included; new instrumentation is a key feature of this book. Because the oxidative stress process involves a cascade of complex, interacting events--for example, (1) generation of the primordial radical, (2) formation of lipid and steroid intermediates that propagate the reaction, (3) modulation of signal transduction pathways by upregulation of regulatory enzymes through nuclear transcription factors and gene expression, and (4) termination by radical inactivation it is clear that more than one method of analysis is often required.

Preface

Table 1 Disorders Associated with Oxidative Stress

v//

Neurological Alzheimers Disease Down's Syndrome Amyotrophic Lateral Sclerosis Schizophrenia Tardine Dyskinesis Parkinson's Disease Huntington's Chorea Ataxia Telangectasia Stroke

Ocular Cataract Age Related Macular Degeneration Retinopathy of Prematurity Light Damage

Endocrine Diabetes Infertility Menopause Thyroiditis Neoplasia Hyperthyroidism Musculocutaneous UV Injury Xeroderma Pigmentosum Duchenne Muscular Dystrophy

Vascular Atherosclerosis Coronary Artery Bypass Grafting Circulatory Shock Cardiac and Peripheral Disease

Hemolytic RBC Fragility Nephrotic Syndrome Chronic Renal Failure Membraneous Glomerulonephritis

Renal Hemochromatosis Anemia Sepsis

Hepatic Hepatitis Fatty Necrosis Fetal Alcohol Syndrome

Autoimmune Rheumatoid Arthritis HIV Systemic Lupus Erythematosis

Pulmonary Respiratory Distress Syndrome Cystic Fibrosis Emphysema Sarcoid Alveolitis Chronic Idiopathic Fibrosis

Gastrointestinal Colitis Acute Pancreatitis Gastric Mucosal Erosion Acute Cholecystitis

Other Conditions Idiosyncratic Drug Reactions Neoplasia Trauma Ischemic Reoxygenatiun Injury Multiple Organ Dysfunction Aging Bloom's Syndrome Transplantation Rejection Air Pollution Toxicity Inflammation Thermal Injury Excessive Exercise Apoptosis Intermittant Claudication Obesity Pre-eclampsia

viii Preface

The reader will find this book a user friendly and helpful resource that is arranged into separate sections related to ROS, LHP, and AOX methodology. Part III describes six procedures that are applicable to both of these functions.

Instructors teaching courses on oxidative stress should find this book equally valuable as an adjunct for the laboratory component. A companion volume on clinical applications is planned for the near future.

I thank the following organizations for financial support and/or the use of institutional facilities during the preparation of this book: Department of Clinical Laboratory Science, University at Buffalo; Cook Institute for Research and Development; Photon Medical Research Center, Hamamatsu University, Japan; Showa University, Japan and the Japanese Ministry of Higher Educa- tion. My special appreciation is extented to Professor John Walker for his assistance in the review process, M. Ohbayashi for technical assistance, and C. Armstrong for secretarial assistance.

Donald A rmstrong

References 1. Michaelis, L. (1946) Am. Scientist34, 573. 2. Mead, J. (1952) SciencellS, 470. 3. Glavind, J. et al. (1949) Experientia 5, 84,85. 4. Dam, H. and Granados, H. (1945) Acyta Physiol. Scand. 10, 162. 5. Tappel, A. (1953) Arch. Biochem. Biophys42, 293. 6. Grishman, R. (1954) Science 119, 623. 7. Harman, D. (1956)jr. Gerontol. 11,298. 8. Yagi, K. (1968) Vitamins39, 105. 9. Armstrong, D. (1994) Free Radicals in Diagnostic Medicine, Plenum, New York.

10. Armstrong, D. et al. (1984) Free Radicals in Molecular Biology, Aging and Disease, Plenum, New York.

11. Books in Print, vol. 6. (1995) R.R. Bowker's Database Publishing Group, New Providence, NJ, p. 2850.

Contents

Preface ............................................................................................................. v Contributors ................................................................................................... xi i i

PART I. TECHNIQUES FOR THE MEASUREMENT OF OXIDATIVE STRESS

1 Oxygen Consumption Methods: Xanthine Oxidase and Lipoxygenase

Is tvan S tad le r ........................................................................................ 3 2 Spin Trapping and Electron Paramagnetic Resonance

Spectroscopy Daniel J. Brackett, Gemma Wallis, Michael F. Wilson,

and Paul B. McCay ......................................................................... 15 3 In Vivo Detection of Free Radicals in Real Time by Low-Frequency

Electron Paramagnetic Resonance Spectroscopy Gerald M. Rosen, Sovitj Pou, and Howard J. Halpern ................... 27

4 Single Photon Counting Yosh ino r i M i z u g u c h i .......................................................................... 37

5 Measurement of Reactive Oxygen Species in Whole Blood and Mononuclear Cells Using Chemiluminescence

Kuldip Thusu, Ehad AbdeI-Rahman, and Paresh Dandona ......... 57 6 Assay of Phospholipid Hydroperoxides by Chemiluminescence-

Based High-Performance Liquid Chromatography Yorihiro Yamamoto, Yasuhiro Kambayashi,

and Takako Ueda ............................................................................ 63

7 Sensitive and Specific Fluorescent Probing of Oxidative Stress in Different Classes of Membrane Phospholipids in Live Cells Using Metabolically Integrated cis-Parinaric Acid

Valerian E. Kagan, Vladimir B. Ritov, Yulia Y. Tyurina, and Vlad im i r A. Tyur in ................................................................... 71

8 Aromatic Hydroxylation: Salicyl ic Ac id as a Probe for Measuring Hydroxyl Radical Production

Andrea Ghiselli ................................................................................... 89

9 Simple Assay for the Level of Total Lipid Peroxides in Serum or Plasma Kunio Yagi ......................................................................................... 101

/x

x

10

Con ten ts

Simple Procedure for Specific Assay of Lipid Hydroperoxides in Serum or Plasma

Kunio Yagi ......................................................................................... 107 11 Antibodies Against Malondialdehyde-Modified Proteins:

Induction and ELISA Measurement of Specif ic Antibodies Yves Chancere l le , J. Mathieu, and J. F. K e r g o n o u ...................... 111

12 Oxidized LDL and Lp(a): Preparation, Modification, and Analysis Jan Galle and Christoph Wanner ................................................... 119

13 Oxidized and Unoxidized Fatty Acyl Esters Odi le Sergent and Josiane Ci l la rd ................................................. 131

14 Synthesis of Lipid and Cholesterol Hydroperoxide Standards Donald Armstrong and Richard W. B rowne .................................. 139

15 Separation of Hydroxy and Hydroperoxy Polyunsaturated Fatty Acids by High-Pressure Liquid Chromatography

Richa rd W. B rowne and Donald Armstrong .................................. 14 7 16 Products of Creatinine with Hydroxyl Radical as a Useful Marker

of Oxidative Stress In Vivo Kazumasa Aoyagi, Sohji Nagase, Akio Koyama,

Mitsuharu Narita, and Shizuo Tojo ............................................ 157 17 Quantitative Analysis of Peptide and Protein Changes in Ischemic

Hippocampal Tissue by HPLC Thomas M. Wengenack, J. Randa l l Slemmon,

and J. Mark Ordy .......................................................................... 165 18 Electrochemical Detection of 8-Hydroxy-2-Deoxyguanosine Levels

in Cellular DNA Thomas M. Nicotera and Sof ia Bard in ........................................... 181

19 Nitric Oxide Synthase Ahmad Aljada and Paresh Dandona .............................................. 191

20 Quantitation of Heine Oxygenase (HO-1) Copies in Human Tissues by Competitive RT/PCR

Nader G. Abraham ............................................................................ 199 21 A Fluorescent-Based Assay for Measurement of Phospholipase A 2

Activity: A Facile Assay for Cell Sonicates

Thomas M. Nico tera and Glenn Spehar ......................................... 211

22 Autofluorescent Ceroid/Lipofuscin Dazhong Yin and UIf B runk ............................................................. 217

23 In Vivo Technique for Autofluorescent Lipopigments Frangois C. Delori and C. Kathleen Dorey .................................... 229

24 An Ex Vivo Erythrocyte Model for Investigating Oxidative Metabolism

Luigia Rossi and Mauro Magnani ................................................... 245

Contents xi

25 Purification of Vesicular Carriers from Rat Hepatocytes by Magnetic Immunoadsorbtion

Lucian Saucan .................................................................................. 257

PART II. TECHNIQUES FOR THE MEASUREMENT OF ANTIOXIDANT ACTIVITY

26 Simultaneous Determination of Serum Retinol, Tocopherols. and Carotenoids by HPLC

Richard W. Browne and Donald Armstrong .................................. 269 27 EPR Measurements of Nitric Oxide-Induced Chromanoxyl Radicals

of Vitamin E: Interactions with Vitamin C Valerian E. Kagan and Nikolai V. Gorbunov ................................. 277

28 Nonvitamin Plasma Antioxidants Nicholas J. Miller .............................................................................. 285

29 Regulatory Antioxidant Enzymes C. E. Pippenger, Richard W. Browne,

and Donald Armstrong ................................................................ 299 30 In Vitro Screening for Antioxidant Activity

Donald Armstrong, Tadahisa Hiramitsu, and Takako Ueda ........ 315 31 Antioxidant Activity of Low-Density Lipoprotein

Nicholas J. Miller and George Paganga ........................................ 325 32 Lipoic Acid as an Antioxidant: The Role of Dihydrolipoamide

Dehydrogenase Mulchand S. Patel and Young Soo Hong ...................................... 337

33 Reduced Glutathione and Glutathione Disulfide Richard W. Browne and Donald Armstrong .................................. 34 7

34 Analysis of Coenzyme Qlo Content in Human Plasma and Other Biological Samples

Scott Graves, Marianna Sikorska, Henryk Borowy-Borowski, Rodney J. H. Ho, Tot Bui, and Clive Woodhouse .................... 353

PART III. TECHNIQUES APPLICABLE TO BOTH OXIDATIVE STRESS AND ANTIOXIDANT ACTIVITY

35 A Simple Luminescence Method for Detecting Lipid Peroxidation and Antioxidant Activity In Vitro

Minoru Nakano, Takashi Ito, and Tadahisa Hiramitsu ................. 369 36 Analysis of Oxidation and Antioxidants Using Microtiter Plates

German Camejo, Boel Wallin, and Mervi Enoj~rvi ....................... 377 37 Trace Element Analysis of Biological Samples by Analytical Atomic

Spectroscopy Hiroki Haraguchi, Eiji Fujimori, and Kazumi Inagaki ................... 389

38 Energy Dispersive X-Ray Microanalysis Don A. Samuelson ............................................................................ 413

xii Contents

39 Pro-Oxidant and Antioxidant Effects of Estrogens Joachim G. Liehr and Deodutta Roy .............................................. 425

40 Artificial Radical Generating and Scavenging Systems: Photo-Fenton Reagent and Caged Compounds

Seiichi Matsugo and Ken Fujimori ................................................. 437 Index ............................................................................................................ 451

Contributors

EHAD ABDEL-RAHMAN ° Molecular Biology Laboratory, Endocrine-Diabetes Center of Western New York, Millard Fillmore Health System, Buffalo, NY

NADER G. ABRAHAM ° Department of Gene Therapy, New York Medical College, Valhalla, NY

AHMAD ALJADA • Endocrinology-Diabetes Center of Western New York, Millard Fillmore Health System, Buffalo, NY

KAZUMASA AOYAGI ° Division of Nephrology, Department of Internal Medicine, Institute of Clinical Medicine, University of Tsukuba, Japan

DONALD ARMSTRONG • Department of Clinical Laboratory Science and Pathology, State University of New York at Buffalo, NY

SOFIA BARDIN " D e p a r t m e n t of Biophysics, Roswell Park Cancer Institute, New York State Department of Health, Buffalo, NY

HENRYK BOROWY-BOROWSK1 • Gloucester, Ontario, Canada DANIEL J . BRACKETT " D e p a r t m e n t of Surgery, College of Medicine,

The University of Oklahoma Health Sciences Center, Oklahoma, City, OK RICHARD W . BROWNE ° Department of Clinical Laboratory Science,

State University of New York at Buffalo, NY ULF BRUNK • Department of Pathology, Faculty of Health Sciences,

University of Linkoping, Sweden TOT BuI • Department of Pharmaceutics, University of Washington, Seattle, WA GERMAN CAMEJO ° Department of Biochemistry, Astra Hassle AB,

Molndal, Sweden YVES CHANCERELLE " CRSSA, Unite de Radiobiochimie, La Tronche, France JOSIANE CILLARD ° Laboratorie de Biologie Cellulaire et Vegetale,

UFR des Sciences Pharmaceutiques, Universitb de Rennes, France PARESH DANDONA ° Department of Medicine, University of Buffalo School

of Medicine and Biomedical Sciences, Endocrinology-Diabetes Center of Western New York, Millard Fillmore Hospital, Buffalo, NY

FRANffOIS C. DELORI • The Schepens Eye Research Institute, Harvard University School of Medicine, Boston, MA

C. KATHLEEN DOREY • The Schepens Eye Research Institute, Harvard University School of Medicine, Boston, MA

xi i i

xiv Contributors

MERVI ENOJ.~,RVI * Department of Biochemistry, Astra Hassle AB, Molndal, Sweden

Em FUJIMORI * Department of Applied Chemistry, School of Engineering, Nagoya University, Japan

KEN FUJIMORI * Department of Chemistry, University of Tsukuba, Japan JAN GALLE * Abt. Nephrologie, Universitatsklinic, Julius Maximilians

Universitat Wurzburg, Germany ANDREA GHISELLI ° Instituto Nazionale della Nutrizione, Roma, Italy NIKOLAI V. GORBUNOV "Depa r tmen t of Respiratory Research, Walter Reed

Army Institute of Research, Washington, DC SCOTT GRAVES • NeoRx Corporation, Seattle, WA HOWARD J. HALPERN • Department of Pharmacology and Toxicology, School

of Pharmacology, University of Maryland at Baltimore, MD HIROKI HARAGUCHI • Department of Applied Chemistry, School

of Engineering, Nagoya University, Nagoya, Japan TADAHISA HIRAMITSU • Photon Medical Research Center, Hamamatsu

University School of Medicine, Hamamatsu, Japan KAZUMI INAGAKI * Department of Applied Chemistry, School of Engineering,

Nagoya University, Nagoya, Japan TAKASHI ITO ° Japan Immunoresearch Laboratories Co., Ltd.,

Takasaki-city, Japan RODNEY HO • Department of Pharmaceutics, University of Washington,

Seattle, WA YOUNG Soo HONG • Department of Biochemistry, School of Medicine

and Biomedical Sciences, State University of New York at Buffalo, NY VALERIAN E. KAGAN ° Department of Environmental and Occupational

Health, University of Pittsburgh Graduate School of Public Health, Pittsburgh, PA

YASUHIRO KAMBAYASHI • Department of Photon and Free Radical Research, Japan Immunoresearch Laboratories Co., Ltd., Takasaki, Japan

J. F. KERGONOU • CRSSA, Unitb de Radiobiochimie, La Tronche, France AKIO KOYAMA • Division of Nephrology, Department of Internal Medicine,

Institute of Clinical Medicine, University of Tsukuba, Japan JOACHIM G. LEHR • Department of Pharmacology and Toxicology,

The University of Texas Medical Branch at Galveston, Galveston, TX MAURO MAGNANI * Instituto di Chimica Biologica "Giorgio Fornaini"

Universita degli Studi di Urbino, Urbino, Italy J. MATHIEU ° CRSSA, Unit~ de Radiobiochimie, La Tronche, France

Contr ibutors xv

SEncm MATSUCO • Department of Chemical and Biochemical Engineering, Faculty of Engineering, Toyama University, Japan

PAUL B. McCAY • National Biomedical Center for Spin Trapping and Free Radicals, Free Radical Biology and Aging Program, Oklahoma Medical Research Foundation, Department of Veterans Affairs Medical Center, Oklahoma City, OK

NICHOLAS J. M]LLER° FRCPath, Free Radical Research Group, Division of Biochemistry and Molecular Biology, UMDS-Guy's Hospital Medical School, London, UK

YOSHINORI MIZUGUCHI • Systems Division, Department 17 Biomedical Applications, Hamamatsu Photonics K.K.d., Hamamatsu, Japan

SOHJ1 NAGASE • Division of Nephrology, Department of Internal Medicine, Institute of Clinical Medicine, University of Tsukuba, Japan

MINORU NAKANO • Japan Immunoresearch Laboratories Co., Ltd., Takasaki-city, Japan

MITSUHARU NARITA • Division of Nephrology, Department of Internal Medicine, Institute of Clinical Medicine, University of Tsukuba, Japan

THOMAS M. NICOTERA ° Department of Biophysics, Roswell Park Cancer Institute and University at Buffalo School of Medicine and Biomedical Sciences, New York State Department of Health, Buffalo, NY

J. MARK ORDY • Department of Neuroscience, John Carroll University, Cleveland, OH

GEORGE PAGANGA ° Free Radical Research Group, Division of Biochemistry and Molecular Biology, UMDS-Guy's Hospital Medical School, London, UK

MULCHAND S. PATEL ° Department of Biochemistry, School of Medicine and Biomedical Sciences, University at Buffalo, NY

CHARLES E. PIPPENGER ° Cook Institute for Research and Education, Butterworth Hospital and Michigan State University School of Medicine, Grand Rapids, MI

SOWTJ PON • Department of Pharmacology and Toxicology, School of Pharmacology, University of Maryland at Baltimore, MD

VLADIMm B. RaTOV • Department of Environmental and Occupational Health, University of Pittsburgh, PA

GERALD M. ROSEN • Pharmaceutical Sciences Department, Pharmacology and Toxicology Program, School of Pharmacy, University of Maryland at Baltimore, MD

LuICIA Rossl • Instituto di Chimica Biologica "Giorgio FornainL '" Universita degli Studi di Urbino, Urbino, Italy

xvi Contributors

DEODUTTA ROY • Department of Pharmacology and Toxicology, The University of Texas Medical Branch at Galveston, TX

DON A. SAMUELSON • Graduate Studies and Research, Department of Small Animal Clinical Sciences, College of Veterinary Medicine, University of Florida, Gainesville, FL

LUCIAN SAUCAN • Vascular Disease Prevention Laboratory, Carl T. Hayden VA Medical Center, Phoenix, AZ

MARIANNA SIKORSKA • Institute for Biological Sciences, National Research Council of Canada, Ottawa, Ontario, Canada

ODILE SERGENT ° Laboratorie de Biologie Cellulaire et Vegetale, UFR des Sciences Pharmaceutiques, Universitk de Rennes, France

J. RANDALL SLEMMON • Department of Biochemistry, University of Rochester, NY GLENN SPEHAR • Department of Biophysics, Roswell Park Cancer Institute,

New York State Department of Health, Buffalo, iVY I S T V A N S T A D L E R • HEMEX Laboratories, Department of Medicine,

State University of New York at Buffalo, Buffalo General Hospital, Buffalo, NY

SHIZUO ToJo • Division of Nephrology, Department of Internal Medicine, Institute of Clinical Medicine, University of Tsukuba, Japan

KULDIP THUSU • Vascular Lab, Endocrinology-Diabetes Center of Western New York, Millard Fillmore Health System, Buffalo, NY

VLADIMIR A. TYURIN • Department of Environmental and Occupational Health, University of Pittsburgh, Pittsburgh, PA

YULIA Y. TYURINA • Department of Environmental and Occupational Health, University of Pittsburgh, PA

TAKAKO UEDA • Department of Pharmacology, Showa University School of Medicine, Tokyo, Japan

BOLL WALLIN • Department of Biochemistry, Astra Hassle AB, Molndal, Sweden

GEMMA WALLIS • Department of Surgery, College of Medicine, The University of Oklahoma Health Sciences Center, Oklahoma City, OK

C H R I S T O P H W A N N E R • Abt. Nephrologie Universitatsklinic, Julius Maximilians-Universitat Wurzburg, Germany

THOMAS M. WENGENACK • Department of Neurology, Mayo Foundation and Clinic, Rochester, MN

M I C H A E L F . W I L S O N • Department of Medicine-Cardiology, Mill~ird Fillmore Health Systems, University at Buffalo School of Medicine and Biomedical Sciences, Buffalo, NY

Contributors xvii

CLIVE WOODHOUSE • Receptagen, Inc., Edmonds, WA KuNio YAGI ° Institute for Applied Biochemistry and Nagoya University

School of Medicine, Yagi Memorial Park, Mitake, Gifu, Japan YORIHIRO YAMAMOTO ° Research Center for Advanced Science

and Technology, University of Tokyo, Tokyo, Japan DAZHONG YIN ° Faculty of Health Sciences, University of Linkoping,

Linkoping, Sweden