Redox Signaling and Regulation inBiology and Medicine

Edited byClaus Jacob and Paul G. Winyard

57268File Attachmentcover.jpg

Redox Signaling and Regulation

in Biology and Medicine

Edited by

Claus Jacob and Paul G. Winyard

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Redox Signaling and Regulation inBiology and Medicine

Edited byClaus Jacob and Paul G. Winyard

The Editors

Jun. -Prof. Dr. Claus JacobUniversität des SaarlandesBioorgan. Chemie-Geb.B21Campus66123 Saarbrücken

Prof. Dr. Paul G. WinyardUniversity of ExeterBiomedical & Clinical ScienceSt. Lukes CampusExeter EX1 2LUUnited Kingdom

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Contents

Preface XVThe Editors XVIIList of Authors XIX

1 Introduction 1Claus Jacob and Paul G. Winyard

2 Biological Systems Relevant for Redox Signaling and Control 13Thomas R. Hurd and Michael P. Murphy

2.1 Introduction 132.2 Reactive Oxygen Species 142.2.1 The Superoxide Radical 142.2.1.1 Generation of the Superoxide Radical 142.2.1.2 The Superoxide Radical as a Redox Signal 162.2.1.3 Decomposition of the Superoxide Radical 172.2.2 Hydrogen Peroxide 192.2.2.1 Generation of Hydrogen Peroxide 192.2.2.2 Mechanisms of Hydrogen Peroxide Signaling 202.2.2.3 Decomposition of Hydrogen Peroxide 272.3 Reactive Nitrogen Species 312.3.1 Nitric Oxide 312.3.1.1 Generation of Nitric Oxide 322.3.1.2 Mechanisms of Nitric Oxide Signaling 322.3.1.3 Decomposition of Nitric Oxide 322.3.2 Peroxynitrite and Reactive Nitrogen Species 332.3.2.1 Generation of Peroxynitrite and Other Important Reactive

Nitrogen Species 332.3.2.2 Mechanisms of Peroxynitrite- and Reactive Nitrogen Species-Mediated

Redox Signaling 34

V

2.4 Lipid Peroxidation Products 362.4.1 Generation of Lipid Peroxidation Products 362.4.2 Mechanisms of Signaling with Lipid Peroxidation Products 372.4.3 Decomposition of Lipid Peroxides 382.5 Conclusions 39

References 40

3 Cellular Generation of Oxidants: Relation to Oxidative Stress 45Lars-Oliver Klotz and Helmut Sies

3.1 Introduction 453.2 Molecular Oxygen and Reactive Oxygen Species: Biochemical

Relations and Endogenous Sources 453.2.1 Endogenous Sources of Superoxide and Superoxide-Derived

Reactive Oxygen Species 463.2.2 Singlet Oxygen 503.2.3 Secondary Reactive Oxygen Species Generated in Radical Chain

Reactions 513.3 Generation of Oxidative Stress Under the Influence of Xenobiotics

and Stressful Stimuli 533.3.1 Quinones and Other Redox Cyclers 533.3.2 Antioxidant Depletion by Alkylation: Acetaminophen Toxicity 563.3.3 Ultraviolet Radiation 573.3.4 Ultrafine or Nanoparticles 57

References 61

4 The Chemical Basis of Biological Redox Control 63Claus Jacob, Mandy Doering, and Torsten Burkholz

4.1 Introduction 634.2 Forms fo Elemental Oxygen as Reactive Oxygen Species 674.2.1 Reactive Oxygen Species and Related Cellular Oxidants 674.2.2 Singlet Oxygen (1O2) 674.2.3 Ozone (O3) 714.3 Reduced, Yet Oxidizing: the Chemistry of Oxygen in Oxidation

States between 0 and �2 724.3.1 Superoxide Radicals (O2

��) 734.3.2 The Superoxide to Peroxide Conversion as a Key Event in Redox

Signaling 764.3.3 Hydrogen Peroxide (H2O2) 774.3.4 Hydroxyl Radicals (HO�) 794.3.5 Enzymatic Reduction of Hydrogen Peroxide by Peroxiredoxins,

Catalase and Glutathione Peroxidase 814.4 The Role of Labile Metal Ions in Oxidative Stress 844.5 Follow-on Species Generated by Chemical Interactions of Reactive

Oxygen Species 85

VI Contents

4.5.1 Hypochlorous Acid (HOCl) 864.5.2 Peroxynitrite (ONOO�) 884.6 Nitrogen Monoxide and Reactive Nitrogen Species 914.6.1 Reactive Nitrogen Species 914.6.2 S-Nitrosothiols 954.7 Sulfur as a Prime Target of Oxidative Stress 984.7.1 Thiol Groups in Peptides and Proteins 994.7.2 The Concept of Reactive Sulfur Species 1014.7.3 Sulfur-Centered Radicals 1034.7.4 Disulfide-S-Oxides (RS(O)xSR0) 1064.7.5 Sulfenic and Sulfinic Acids (RSOH and RS(O)OH) 1084.7.6 Oxidation of Methionine: Sulfoxides (RS(O)R) and Sulfones

(RS(O)2R0) 1094.7.7 S-Thiolation as Chemical Protection 1104.7.8 Oxidation of Disulfides 1114.8 A Brief Overview of Hydroxylation and Nitration

Reactions 1124.8.1 Hydroxylation and Nitration of Aromatic Residues 1134.8.2 Fatty Acid Chemistry 1164.9 Conclusion 117

References 118

5 Protein Glutathiolation 123Pietro Ghezzi and Paolo Di Simplicio

5.1 Introduction: Glutathione – From Antioxidant to RedoxSignal 123

5.2 Glutathiolation 1245.3 Mechanisms of Glutathiolation 1275.3.1 ROS-Dependent Glutathiolation by Thiol/Disulfide Exchange

Reactions 1295.3.2 ROS-Dependent Glutathiolation via Sulfenic Acid 1305.3.3 ROS-Dependent Glutathiolation by Radical

Reactions 1325.3.4 ROS-Independent Glutathiolation 1335.4 Toxicological Aspects of Glutathiolation 1335.5 Mechanisms of Deglutathiolation 1355.6 Enzymes Implicated in Glutathiolation 1365.7 Specificity of Glutathiolation 1365.8 Genetic Factors Affecting Glutathiolation 1375.8.1 Mouse Beta-Globin 1375.8.2 Glucose-6-Phosphate Dehydrogenase Mutations 1375.8.3 Glutathione S-Transferase Mutations 1385.9 Future Perspectives 138

References 139

Contents VII

6 Structure and Function of the Human Peroxiredoxin-BasedAntioxidant System: the Interplay between Peroxiredoxins,Thioredoxins, Thioredoxin Reductases, Sulfiredoxins andSestrins 143Katalin É. Szabó, Kirsty Line, Paul Eggleton, Jennifer A. Littlechild,and Paul G. Winyard

6.1 Introduction 1436.2 Peroxiredoxins 1476.3 Thioredoxins 1586.4 Thioredoxin Reductases 1596.5 Sulfiredoxin and Sestrins 1616.6 Regulation of the Expression and Activity of the Peroxiredoxin-Based

System Enzymes 1636.7 Cell Signaling and the Peroxiredoxin-Based System: Regulation of

Transcription Factors, Cell Cycle and Apoptosis 1666.8 Peroxiredoxin-Based System in Cells and Organs of the Body 1706.9 Summary 173

References 174

7 Hydrogen Peroxide and Cysteine Protein Signaling Pathways 181Ewald Schröder and Philip Eaton

7.1 Introduction 1817.2 Hydrogen Peroxide Production in Cells and Tissues 1827.3 Sources and Concentrations of Hydrogen Peroxide 1837.4 Hydrogen Peroxide and the Formation of Secondary Oxidants such

as Peroxynitrite 1847.5 Exogenous Hydrogen Peroxide as an Experimental Tool 1867.6 Hydrogen Peroxide Sensing and Cysteine Sensors 1887.7 Low Molecular Weight Oxidized Thiols and S-Thiolated Protein

Adducts 1887.8 Protein Thiol Modifications 1897.9 Protein Regulation via Sulfination-Dependent Proteolysis 190

References 192

8 Protein Tyrosine Phosphatases as Mediators of Redox Signaling 197Jeroen den Hertog

8.1 Introduction 1978.2 Protein Tyrosine Phosphatases 1988.3 Protein Tyrosine Phosphatases are Sensitive to Oxidation 1998.4 Allosteric Regulation of Receptor Protein Tyrosine Phosphatases

by Oxidation 2008.5 Activation of Sdp1 by Oxidation 2028.6 Physiological Relevance of Protein Tyrosine Phosphatase Oxidation 2038.7 Conclusions 204

References 204

VIII Contents

9 Hypoxia-Induced Gene Regulation through Hypoxia InducibleFactor-1a 207Adam J. Case and Frederick E. Domann

9.1 Introduction 2079.2 The Proteins and Mechanism 2089.3 Hypoxia Inducible Factor Target Genes 2139.4 Non-Hypoxia-Induced Activation of Hypoxia Inducible

Factor 2189.5 Diseases Involving Hypoxia Inducible Factor 2209.6 The Promise of Hypoxia Inducible Factor-Targeted Therapies 2239.7 Conclusions 226

References 226

10 Eicosanoid-Based Signaling 229Valerie B. ODonnell

10.1 Introduction 22910.2 Biosynthesis and Structures of Eicosanoids 23110.2.1 Lipoxygenases 23410.2.2 Cyclooxygenases 23510.2.3 Cytochrome P450 23710.3 Signaling by Eicosanoids 23910.4 Metabolism of Eicosanoids 24010.5 Summary 242

References 243

11 Redox-Controlled Transcription Factors and Gene Expression 245Gregory I. Giles

11.1 Introduction 24511.2 Redox Signaling and Gene Microarry Data: The Global

Picture 24811.3 The Antioxidant Response Element 25111.4 The Transcription Factor Nrf2: The Master Regulator of Antioxidant

Transcription 25311.5 The Keap1–Nrf2 Complex: A Sensor for Cellular Stress 25511.5.1 Reactive Oxygen Species, Reactive Nitrogen Species and Electrophile

Sensing by Keap1 25611.5.2 Structural Basis for the Sensing Function of the Keap1–Nrf2

Complex 25711.6 Redox Reactions of Transcription Factors 26111.6.1 Nuclear Factor kB: Redox Control of the Immune Response 26211.6.2 Activator Protein 1 (AP-1): Redox Control of Proliferation and

Apoptosis 26411.6.3 The Role of the Nuclear Redox State in Gene Expression 26511.7 Summary 266

References 267

Contents IX

12 Nitric Oxide Regulation in Redox Signaling 271Dario A. Vitturi, David M. Krzywanski, Edward M. Postlethwait, andRakesh P. Patel

12.1 Introduction 27112.2 Nitric Oxide Formation 27112.3 Factors Affecting Nitric Oxide Reactivity and Signaling 27212.3.1 Compartmentalization and Diffusion 27212.3.2 Interaction with Metal Centers 27412.3.3 Nitric Oxide Reaction with Free Radicals 27612.3.3.1 Nitric Oxide and Superoxide 27612.3.3.2 Peroxynitrite 27812.3.3.3 Nitrogen Dioxide/Dinitrogen Trioxide 28112.4 Nitric Oxide Signaling Beyond Soluble Guanylate Cyclase 28112.4.1 Nitric Oxide and Mitochondria 28212.4.2 S-Nitrosation 28212.4.3 Nitrated Lipids 28312.4.4 3-Nitrotyrosine 28412.5 Redox Derivatives of Nitric Oxide 28412.5.1 Nitroxyl Anion 28512.5.2 Nitrite 28512.5.3 Nitrite – a Potential Reservoir for Nitric Oxide Bioactivity

During Hypoxia 28612.6 Summary 287

References 287

13 Is Hydrogen Sulfide a Regulator of Nitric Oxide Bioavailabilityin the Vasculature? 293Matthew Whiteman and Philip K. Moore

13.1 Introduction 29313.2 Reactive Nitrogen Species 29313.3 Reactive Nitrogen Species in the Heart and Vasculature 29513.4 Hydrogen Sulfide Biosynthesis 29613.5 Hydrogen Sulfide Measurement, Catabolism and Removal 30013.6 Hydrogen Sulfide in the Heart and Vasculature 30513.7 What is the Evidence for Crosstalk between Nitric Oxide

and Hydrogen Sulfide 30913.8 Nitric Oxide/Hydrogen Sulfide and Evidence for the Formation

of A Novel Intermediate 31213.9 Concluding Remarks 313

References 314

14 Aspects of Nox/Duox Signaling 317Masuko Ushio-Fukai

14.1 Introduction 31714.2 The Nox/Duox Enzymes (Expression and Domain Structure) 318

X Contents

14.2.1 NOXO1 and NOXA1 32214.3 Mechanism of the Nox/Duox Activation 32214.3.1 Nox1 32314.3.2 Nox3 32414.3.3 Nox4 32414.3.4 Nox5 32414.3.5 Duox 32514.4 Redox Signaling Activated by NADPH Oxidase 32514.4.1 Activation of Kinases and Phospholipases 32614.4.2 Activation of Ion Channels and Calcium Signaling 32714.4.3 Oxidative Inactivation of Protein Tyrosine Phosphatase 32814.4.4 Specific Localization of NADPH Oxidase as Mechanism of Activation

of Specific Redox Signaling 32814.5 Transcription Factors and Gene Expression Regulated by ROS 33014.5.1 NF-kB 33114.5.2 AP-1 (c-Jun and c-Fos) 33114.5.3 HIF-1 33114.5.4 Ets 33214.5.5 p53 33214.6 Functional Role of Nox/Duox in Physiological and Pathophysiological

Functions 33314.6.1 Function of Nox1 33314.6.2 Function of Nox2 33314.6.3 Function of Nox3 33614.6.4 Function of Nox4 33614.6.5 Function of Nox5 33914.6.6 Function of Duox 34014.7 Summary and Conclusions 340

References 341

15 Photodynamic Therapy with Aminolevulinic Acid and Iron Chelators:A Clinical Example of Redox Signaling 351Andrew Pye, Yuktee Dogra, Jessica Tyrrell, Paul Winyard, andAlison Curnow

15.1 Photodynamic Therapy 35115.2 The Development of Photodynamic Therapy 35215.3 Aminolevulinic Acid Photodynamic Therapy 35315.4 Heme Biosynthesis and Regulation During ALA-PDT 35415.5 Iron and the Enhancement of ALA-PDT 35615.6 Redox Signaling in Photodynamic Therapy after Light

Irradiation 36115.7 Photosensitizers as Reactive Oxygen Species Generators in

Redox Research 36315.8 Reactive Oxygen Species Generation and Signaling in Clinical

Photodynamic Therapy 364

Contents XI

15.9 Apoptosis in Photodynamic Therapy 36415.10 Subcellular Localization of Reactive Oxygen Species

Production 36515.11 Changes in Transcription Factor and Protein Levels Following

Photodynamic Therapy 36615.12 Iron and Iron Chelation Post Light Activation of

Photosensitizer 36715.13 Vascular Damage, Hypoxia and Hypoxia Inducible Factor 36815.14 Conclusion 369

References 370

16 Oxidative Stress and Apoptosis 373Silvia Cristofanon, Mario Dicato, Lina Ghibelli, and Marc Diederich

16.1 Apoptosis, The Programmed Destiny of a Cell 37316.2 Historical Overview of the Relation Between Oxidative Stress and

Apoptosis 37816.3 Oxidative Stress as a Mediator and Inducer of Apoptosis 378

References 382

17 Redox Regulation of Apoptosis in Immune Cells 385Edith Charlier, Jacques Piette, and Geoffrey Gloire

17.1 Apoptosis, Necrosis and Autophagy 38517.2 Apoptosis 38617.2.1 Morphological and Biochemical Features of Apoptosis 38617.2.2 Molecular Mechanisms of Apoptosis 38617.2.2.1 The Major Players in Apoptosis 38617.2.2.2 Main Pathways of Apoptosis 39217.3 Redox Regulation of Apoptosis in Immune Cells 39517.3.1 Induction of Apoptosis by Exogenous Reactive Oxygen Species 39617.3.1.1 Reactive Oxygen Species Targets 39617.3.1.2 Reactive Oxygen Species-Induced Apoptosis in Immune Cells 39717.3.2 Control of Apoptosis by Endogenous Reactive Oxygen Species

Production 39817.3.2.1 Cytokine Stimulation 39817.3.2.2 Immune Receptor Stimulation 39917.3.2.3 Granzyme A Delivery 40217.3.2.4 Spontaneous and Bacteria-Induced Neutrophil Apoptosis 403

References 405

18 Redox Control in Human Disease with a Special Emphasis on thePeroxiredoxin-Based Antioxidant System 409Katalin É. Szabó, Nicholas J. Gutowski, Janet E. Holley, Jennifer A.Littlechild, and Paul G. Winyard

18.1 Introduction 40918.2 Inflammatory Diseases 410

XII Contents

18.3 Neurodegenerative Diseases 41418.4 Diseases of the Eye 41818.5 Blood Disorders 41918.6 Atherosclerosis and Cardiovascular Disease 42018.7 Infections and Parasitic Diseases 42118.8 Cancer 42218.9 Proteins of the Peroxiredoxin-Based System as Diagnostic

and Prognostic Tools, and Potential Drug Targets 42418.10 Conclusions 426

References 427

19 Free Radicals and Mammalian Aging 433Alberto Sanz, Gustavo Barja, Reinald Pamplona,and Christiaan Leeuwenburgh

19.1 Introduction 43319.2 The Mitochondrial Free Radical Theory of Aging 43419.3 Why are Long-Lived Animals so Long-Lived? Four Correlations

to Explain Longevity 43519.3.1 Mitochondrial Free Radical Generation 43619.3.2 Is Complex I the Major Determinant of Aging Rate? 43819.3.2.1 Decrease in the Concentration of Respiratory Complexes 44019.3.2.2 Modification of the Amino Acid Composition of Respiratory

Subunits 44019.3.2.3 Posttranslational Modification of Specific Subunits 44119.3.2.4 Changes in mtROS Production Without Altering the Electron

Transport Chain 44219.3.3 The Unsaturation Degree of Membrane Fatty Acids 44219.3.4 Methionine Content in Proteins 44319.3.5 G þ C Content in mtDNA 44419.3.6 In Summary: Why are Long-Lived Animals So Long-Lived? 44419.4 Oxidative Damage to Macromolecules 44619.4.1 Oxidative Damage to Carbohydrates 44619.4.2 Oxidative Damage to Lipids 44719.4.3 Oxidative Damage to Proteins 44819.4.4 Oxidative Damage to DNA 44919.4.5 The Importance of Mitochondrial DNA 45019.5 Effects of Mitochondrial DNA Mutations on Cellular Function 45219.5.1 Are There Exponential Increases in Free Radical Production

With Age? 45219.5.2 Apoptosis 45319.5.3 ATP Production 45419.6 Trying to Increase the Maximum Life Span 45619.6.1 Antioxidants: The Great Hope, The Great Deception 45619.6.2 Dietary Restriction: The Cheapest, The Best 45819.6.3 Dietary Restriction and Reduction in Oxidative Stress 458

Contents XIII

19.6.4 How are Mitochondrial ROS Regulated During DietaryRestriction? 460

19.6.4.1 Hormonal Regulation 46119.6.4.2 Regulation of Mitochondrial ROS Production by Specific Dietary

Components: The Role of the Dietary Proteins and Methionine 462References 464

Index 473

XIV Contents

Preface

During the last decade, the area of intracellular redox processes, signaling andcontrol has witnessed many remarkable developments. Discoveries such as thewidespread occurrence of highly oxidized cysteine residues in proteins, sulfiredoxin,sulfinic acid switches in peroxiredoxins, widespread glutathiolation in proteins,redox-controlled chaperone activity, redox-regulated transcription factor activation,antioxidant response networks and redox-controlled apoptotic pathways, haveresulted in a wealth of new knowledge. At the same time, the significance of redoxcontrol has been further underlined by the emerging links between cellular redoxevents and many human illnesses, as well as an apparent connection betweenoxidative stress and aging.Many of these developments have been the focus of recent scientific meetings,

such as the 2006Mosbach Kolloquium in Germany (‘‘Redox Signaling: Mechanismsand biological impact’’) and a high profile Gordon Conference in the USA inJune 2006 (‘‘Thiol-based redox regulation & signaling’’). There are also a numberof excellent reviews, some of which have been authored by the contributors of thisbook, which focus on individual issues of intracellular redox events, for instanceoxidative stress, biological sulfur chemistry, intracellular redox pathways, apoptosisand aging. Considering the present literature, various outstanding books are avail-able, which touch on aspects of cellular redox control, such as free radicals or cellularsignaling pathways. To date, no appropriate book exists, however, which covers thehighly multidisciplinary topics of intracellular redox events in a comprehensiveapproach – and, at the same time, is accessible to students and cutting-edgeresearchers alike.The purpose of this book is therefore four-fold. First, it aims to provide an up-to-

date text that covers the range of recent developments in the area of redox signalingand control. The topics covered will, of course, represent a fine selection of keyissues only, without any claims to complete coverage of this rather extensive area ofresearch. Here, it is hoped that the themes covered will ultimately stimulate furtherreading, and plenty of references are provided throughout to guide the search forliterature. Second, the book aims at a deeper understanding of redox signaling andcontrol events by taking a holistic, cross-discipline approach which combines the

XV

various chemical, analytical, biochemical, biological and medical aspects relevant toredox signaling. In doing so, the book hopes to fulfil its third purpose – to provide aneasy-to-understand text which can be used by research students, postdoctoral fellowsand experienced researchers alike. Here, we have added a set of Explanatory Boxes toeach chapter which will provide a basic, easy-to-understand background for some ofthe more complicated or specialized concepts discussed. While these boxes aredesigned to assist readers in the first instance, they should not been seen ascomprehensive coverage of a particular topic and cannot replace more solid back-ground reading. Last but not least, we hope to provide a book for chemists,biochemists, biologists, oxidative stress and aging researchers that is educational,stimulating and interesting to read.We would like to thank all of our colleagues who have walked with us on this

project, all leading researchers in their fields – without their contributions this bookwould not, of course, have been possible. Special thanks also go to Wiley-VCH, andin particular to Ms. Stefanie Volk and Dr. Frank Weinreich, for their continuoussupport, help, advice and above all patience with this book.

Saarbrücken and Exeter Claus Jacob and Paul WinyardAugust 2008

XVI Preface

The Editors

Claus Jacob is Junior Professor of Bioorganic Chemistry at theSchool of Pharmacy, University of Saarland, Germany. Hestudied chemistry in Kaiserslautern, Germany, and Leicester,UK, subsequently earning his doctorate from the Universityof Oxford. From 1996 until 1999 he was a postdoc underProf. Bert Vallee at Harvard Medical School, before takingup the position of lecturer in inorganic chemistry at theUniversity of Exeter, where he remained until moving to hiscurrent position in 2005. Dr. Jacob is currently coordinator ofthe EU Framework 7 Marie Curie Initial Training Network on‘‘Natural Products and related Redox Catalysts: BasicResearch and Applications in Medicine and Agriculture’’.His particular interest is in the chemistry underlying bio-chemical redox events, most notably reactive sulfur species.Claus Jacob, Universität des Saarlandes, School of Phar-

macy, Division of Bioorganic Chemistry, 66041 Saarbrücken,Germany

Paul G. Winyard has been Professor of Experimental Medi-cine at Peninsula College of Medicine and Dentistry, Exeter,UK, since 2002. He previously held a chair in experimentalmedicine at St Bartholomews and the Royal London School ofMedicine and Dentistry, London, UK, and was a visiting pro-fessor at theUniversity of California, San Francisco, from 2000to 2001. Professor Winyards research interests center on therole of oxidative/nitrosative stress in such chronic inflamma-tory diseases as rheumatoid arthritis, with a particular focus onthe development of novel therapeutic strategies and free radi-cal assays, as well as the translation of these developments intopre-clinical and early-phase clinical studies.Paul G. Winyard, Universities of Exeter and Plymouth,

Peninsula Medical School, St Lukes Campus, Exeter EX12LU, UK

XVII

List of Authors

Gustavo Barja obtained his PhD in biological sciences fromthe Complutense University of Madrid in 1981. He continuedhis scientific research, performing short stays at the Univer-sities of Paris VII (France), UCLA (USA), Brescia (Italy) andETH (Switzerland). He has dedicated his research activities tothe study of free radicals and oxidative stress in animals,focusing on aging and longevity. He is currently full professorof physiology at Complutense University, where he teachescomparative animal physiology to biology students, as well asPhD courses on comparative biochemistry and aging. He haspublished three books and over 150 peer-reviewed papers, hasparticipated in 17 multiyear national or international researchprojects, and has received five prizes for his research intoaging, including the Pfizer Foundation Award in 2000. In2001 he was nominated as a member of the Royal Academy ofPharmacy of Spain.Gustavo Barja, Complutense University, Faculty of Biology,

Department of Animal Physiology-II, Madrid 28040, Spain

Torsten Burkholz studied chemistry at the University of Saar-land, Saarbruecken, Germany. He obtained his first degree inchemistry in June 2006. In October 2006, he joined the Divi-sion of Bioorganic Chemistry and is currently conductingresearch as part of his PhD thesis entitled ‘‘Oxidative stressand electrochemical procedures for surface decontaminationin dialysis’’. This project is supported by Fresenius MedicalCare, Bad Homburg, Germany.Torsten Burkholz, Universität des Saarlandes, School

of Pharmacy, Division of Bioorganic Chemistry, 66041Saarbrücken, Germany

XIX

Adam J. Case is a fifth-year combined MD/PhD student at theUniversity of Iowa. He is currently pursing a PhD in freeradical and radiation biology, and his research interests includeelucidation of the role of antioxidant enzymes in cancer, epi-genetics of regulatory enzymes, and the use of antidiabetesmedication in the prevention of oncogenesis. Following histhesis dissertation, Adam plans to pursue a medical residencyin pediatrics where he hopes to practice both clinical andresearch medicine in the field of pediatric oncology.Adam J. Case, The University of Iowa, Holden Compre-

hensive Cancer Center, Carver College of Medicine, Depart-ment of Radiation Oncology, Iowa City, IA 52242-1181, USA

Edith Charlier is a PhD student at the GIGA-Research Center,Liège, Belgium, in the division of Jacques Piette in the sub-group of Geoffrey Gloire. She studied biochemistry at theUniversity of Liège and she joined the division of JacquesPiettefor her thesis. She isworking on an inositol phosphatasewhichinfluences the sensitivity of T lymphocytes to CD95-inducedcell death.Edith Charlier, University of Liège, Virology-Immunology

Unit, GIGA-Research, 1, avenue de lHopital, 4000 Liège,Belgium

Silvia Cristofanon received her PhD from the University ofRome Tor Vergata in spring 2008 after three years of researchat the LBMCC Lab in Luxembourg. Her major interests arerelated to apototic cell signaling pathways in human leukemiacells induced by glutathione depletion.Silvia Cristofanon, Hôpital Kirchberg, Fondation Recherche

sur le Cancer et les Maladies du Sang, Laboratoire de BiologieMoléculaire et Cellulaire du Cancer (LBMCC), 9, rue EdwardSteichen, 2540 Luxembourg and Universita di Roma ‘‘TorVergata’’, Dipartimento di Biologia, via Ricerca Scientifica,00133 Roma, Italy

Alison Curnow As medical research director of Cornwall Der-matology Research (Peninsula Medical School, UK), AlisonCurnow has established one of the top dermatological photo-dynamic therapy groups in the UK. The clinical and experi-mental research program aims to improve the effectivenessand diversify the application of this noninvasive light-mediatedtreatmentmodality. In addition, the laboratory also investigatesthe causation and potential prevention of skin carcinogenesis.

XX List of Authors

She is currently active in undergraduate teaching andpostgraduate supervision, as well as being a director of theInternational Photodynamic Association Board, and hasnumerous peer-reviewed publications in internationalscientific and clinical journals to her name.Alison Curnow, Cornwall Dermatology Research, Penin-

sula Medical School, The Knowledge Spa, Truro, CornwallTR1 3HD, UK

Jeroen den Hertog received his PhD at Utrecht University in1992, based on a thesis that he prepared at the HubrechtLaboratory, Netherlands Institute for Developmental Biology.He carried out his postdoc under Tony Hunter at the SalkInstitute, La Jolla, CA, USA, from 1992 to 1994. In 1994 hebecame project leader at the Hubrecht Laboratory and in 1997group leader, while also being appointed professor of molecu-lar developmental zoology at the University of Leiden in 2008.Dr den Hertog has over 70 scientific publications to his nameand is the recipient of several grants from the Dutch CancerSociety, Netherlands Organization for Scientific Research,Netherlands Proteomics Centre and the European Commis-sion. His research interest is the role of tyrosine phosphoryla-tion and dephosphorylation in development and disease.Jeroen den Hertog, Hubrecht Institute, Uppsalalaan

8, 3584 CT Utrecht, The Netherlands

Mario Dicato is head of InternalMedicine and of the Service ofHaematology-Oncology at Luxembourg Medical Centre. Muchof his postgraduate training was spent at the University ofPittsburgh, Pittsburgh, Pennsylvania, USA and at HarvardUniversity, Boston, Massachusetts, USA.Mario Dicato, Hôpital Kirchberg, Fondation Recherche sur

le Cancer et les Maladies du Sang, Laboratoire de BiologieMoléculaire et Cellulaire du Cancer (LBMCC), 9, rue EdwardSteichen, 2540 Luxembourg, Luxembourg

Paolo Di Simplicio is professor of pharmacology at the Uni-versity of Siena. His research interest is focused on the func-tional antioxidant aspects of glutathione and other thiols withspecial emphasis on the regulation of the redox state ofbiological systems.Paolo Di Simplicio, University of Siena, Department of

Neuroscience, Pharmacology Unit, 53100 Siena, Italy

List of Authors XXI

Marc Diederich received his PhD in molecular pharmacologyin 1994 at the University of Nancy (France). He is currentlyleading the Laboratoire de Biologie Moléculaire et Cellulairedu Cancer (LBMCC) in Luxembourg. Research in this labora-tory is mainly focused on the inhibition of glutathione-baseddrug resistance mechanism by natural compounds as well aserythroid differentiation mechanisms.Marc Diederich, Hôpital Kirchberg, Fondation Recherche

sur le Cancer et les Maladies du Sang, Laboratoire de BiologieMoléculaire et Cellulaire du Cancer (LBMCC), 9, rue EdwardSteichen, 2540 Luxembourg, Luxembourg

Mandy Doering studied chemistry at the University of Saar-land, where she received her first degree in 2007. She joinedClaus Jacobs group in bioorganic chemistry as part of herundergraduate project and started to work on multifunc-tional, biologically active redox catalysts. After graduation,Mandy continues to work on the design of multifunctionalredox catalysts with regard to potential applications in medi-cine and agriculture in Claus Jacobs research group.Mandy Doering, Universität des Saarlandes, School of Phar-

macy, Division of Bioorganic Chemistry, 66041 Saarbrücken,Germany

Yuktee Dogra obtained a BSc (Hons) in microbial and cellularbiology and a MRes in aquatic ecotoxicology from the Uni-versity of Plymouth. She is currently in her third year ofstudying for a PhD in clinical and biomedical science at thePeninsula Medical School. Her project considers the mechan-isms and enhancement of photodynamic therapy in the areaof dermatology and employs a variety of biological and che-micalmethodologies. She has two published papers to date onthe effects of tritium on the aquatic environment and repre-sents the Schools postgraduate students on the researchdegree committee.Yuktee Dogra, Cornwall Dermatology Research, Peninsula

Medical School, The Knowledge Spa, Truro, Cornwall TR13HD, UK

XXII List of Authors

Frederick E. Domann is a professor of radiation oncology inthe Free Radical & Radiation Biology Program at The Uni-versity of Iowa in Iowa City, Iowa, USA. He earned his PhDat the University of Wisconsin-Madison in human cancerbiology and subsequently carried out postdoctoral researchat The Arizona Cancer Center. He joined the faculty at TheUniversity of Iowa in 1993 and has since become an inter-nationally recognized expert in free radical biology andcancer. His research interests include transcriptional regu-lation and epigenetic control of gene expression in humancancer.Frederick E. Domann, The University of Iowa, Holden

Comprehensive Cancer Center, Carver College of Medicine,Department of Radiation Oncology, Iowa City, IA 52242-1181,USA

Philip Eaton gained a BSc in biochemistry from QueenMary College (University of London) in 1989 before com-pleting his PhD studies at the University of Sussex. Afterpostdoctoral work at the Institute of Psychiatry, he joinedthe Department of Cardiovascular Research, at the RayneInstitute, St. Thomas Hospital in 1995. He remains at theRayne Institute and is currently based in the Department ofCardiology. A major focus of his work is the covalent modi-fication of cardiac proteins by oxidants, with a particularemphasis on thiol-targeted events such as S-thiolation,interprotein disulfide formation, nitrosylation, sulfenationand sulfination.Philip Eaton, Kings College London, Cardiovascular Divi-

sion, Department of Cardiology, The Rayne Institute, St.Thomas Hospital, London SE1 7EH, UK

Paul Eggleton runs the inflammation and autoimmunitygroup at Peninsula Medical School, University of Exeter,Devon, UK. His interest in the biochemistry of inflammatorypathways developed while studying for his PhD at the RoyalCollege of Surgeons of England in London. His interests inleukocyte biology continued at Boston University MedicalSchool, where he demonstrated that oxidative stress proteinsbind to complement proteins and influence innate immunity.His interest in oxidative stress and immunity developedfurther at Oxford University, where his group focused onthe role of complement protein interaction with the oxidativestress protein—calreticulin, and demonstrated that release of

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this protein during cell stress modulated autoimmunity andapoptosis. This work is ongoing at Peninsula Medical School.Paul Eggleton, Universities of Exeter and Plymouth, Penin-

sula Medical School, St. Lukes Campus, Magdalen Road,Exeter EX1 2LU, UK

Pietro Ghezzi, PhD, heads the Laboratory of Neuroimmunol-ogy at the Mario Negri Institute in Milano. His main researchinterests are on the role of inflammatory cytokines in diseasesof the central nervous system and the redox regulation ofimmunity with a particular focus on glutathiolation and redoxproteomics.Pietro Ghezzi, Brighton & Sussex Medical School, Trafford

Centre, Falmer, Brighton BN1 9RY, UK

Lina Ghibelli gained her laurea cum laude in biology in 1979,folowed by postdoctoral training at the University of Chicagoand EMBL in Heidelberg. Since 1991 she has been leading aresearch group studying the molecular mechanisms regulat-ing the process of cell death by apoptosis.Lina Ghibelli, Universita di Roma ‘‘Tor Vergata’’, Diparti-

mentodi Biologia, via Ricerca Scientifica, 00133 Roma, Italy

Gregory I. Giles received his PhD from the University ofSouthampton (UK) and gained postdoctoral experience as aresearch fellow at the University of Exeter (UK). He wasawarded a Frederick Gardner Cottrell Career EnhancementAward to study at the Center for Free Radical Biology at theUniversity of Alabama at Birmingham (USA), followed by hisfirst faculty position as a university research fellow at theUniversity of Sydney (Australia). He was appointed to a lec-tureship at the University of Otago, Dunedin (New Zealand)in 2008. His research interests include drug design, freeradical biology and signal transduction mechanisms.Gregory I. Giles, Department of Pharmacology, Otago

School of Medical Sciences, University of Otago, Dunedin,New Zealand

XXIV List of Authors

Geoffrey Gloire is a postdoctoral researcher from the FNRS(Belgian Fund for Scientific Research) at the GIGA-ResearchCenter, Liège, Belgium, in the laboratory of Jacques Piette. Hestudied biology at the University of Liège and joined thelaboratory of Jacques Piette to complete his PhD on themechanism of NF-kB activation by reactive oxygen species.He now works with his group on the regulation of apoptosisby protein phosphatases and reactive oxygen species inimmune cells.Geoffrey Gloire, University of Liège, Virology-Immunology

Unit, GIGA-Research, 1, avenue de lHopital, 4000 Liège,Belgium

Nicholas J. Gutowski is a consultant neurologist at theRoyal Devon and Exeter Foundation Hospital and seniorlecturer in the Peninsula Medical School. He has long-stand-ing research interests with publications in several aspects ofneuroscience in neurological diseases, including oxidativestress systems, astrocyte and endothelial cell phenotypechanges and neuro-oncology as well as interests in neurode-velopment, in particular the congenital cranial dysinnervationdisorders.

Janet E. Holley completed a BSc in health science at ExeterUniversity in 1998 and an MSc in advanced neuro- andmolecular pharmacology at Bristol University in 1999, whereshe was involved in a research project investigating theantioxidant status of ventilated premature babies. She hasbeen a member of the research team at the Peninsula MedicalSchool in Exeter since its foundation and has recently com-pleted her PhD. Her primary interests are in the biology andfunctions of astrocytes and their role in neuropathology, theinteractions between astrocytes and endothelial cells, oxida-tive stress and antioxidant systems in neurological disease,and congenital cranial dysinnervation disorders.

Thomas R. Hurd, Medical Research Council DunnHumanNutrition Unit, Wellcome Trust/Medical Research CouncilBuilding, Hills Road, Cambridge CB2 0XY, UK

List of Authors XXV

Lars-Oliver Klotz has been an associate professor at theEnvironmental Health Research Institute at Heinrich-Heine-University in Düsseldorf, Germany, since 2007. He studiedbiochemistry at the University of Tübingen, and received hisPhD in biochemistry from the University of Düsseldorf in1998. Following postdoctoral studies at the National Instituteon Aging in Baltimore, MD, USA, he returned to Düsseldorfin 2000, where he received his lecturing qualification in 2001.Dr. Klotz is a recipient of the Catherine-Pasquier-Award of theEuropean Society of Free Radical Research. His researchinterests include the biochemistry of oxidative stress, stress-induced signal transduction and molecular processes inaging.Lars-Oliver Klotz, Department of Molecular Aging

Research at Institut für umweltmedizinische Forschung(IUF), 40225 Düsseldorf, Germany

David M. Krzywanski received his PhD in environmentalhealth science in 2006 from the University of Alabama, Bir-mingham, USA. His research centers on free radical biologyand human health and disease, with his interests rangingfrom the enzymatic regulation of glutathione synthesis to theeffect of environmental nitrogen dioxide on acute epithelialcell injury. Currently, Dr Krzywanski is a postdoctoral fellowin the UAB Department of Pathology, working under Dr S.Ballinger on projects aimed at understanding the role ofmitochondrial signaling in the development of cardiovasculardisease.DavidM. Krzywanski, University of Alabama at Birmingham,

Department of Center for Free Radical Biology, Birmingham,AL, USA

Christiaan Leeuwenburgh received his PhD from the Uni-versity of Illinois, Urbana-Champagne in 1995. He completedhis postdoc in internal medicine at the Division of Geriatricsand Gerontology and Division of Atherosclerosis, Nutritionand Lipid Research at Washington University School of Med-icine, Saint Louis. He became an assistant professor at theUniversity of Florida in 1998, where he is currently a profes-sor within the Department of Aging and Geriatric Research,College of Medicine and Institute on Aging, as well as head-ing the Division of Biology of Aging. Dr Leeuwenburghsmajor research focus is on understanding the molecularmechanism of oxidative stress and apoptosis with age inrodent models.

XXVI List of Authors

Christiaan Leeuwenburgh, University of Florida, College ofMedicine, Division of Biology of Aging, Department of Agingand Geriatrics, Institute on Aging, Biochemistry of AgingLaboratory, Gainesville, FL 32610-0107, USA

Kirsty Line graduated in biochemistry from the University ofWales, Aberystwyth, in 2000. She was awarded her PhD in2004 in the group of Professor Jenny Littlechild, University ofExeter, investigating novel gamma-lactamase enzymes impor-tant in industrial biotransformation reactions, in conjunctionwithChirotech Technology Ltd, Cambridge. She has continuedto work with Professor Littlechild, carrying out studies onhuman antioxidant enzymes peroxiredoxin and sulfiredoxin.Kirsty Line, Universities of Exeter and Plymouth, Peninsula

Medical School, St Lukes Campus, Magdalen Road, ExeterEX1 2LU, UK

Jennifer A. Littlechild carried out her PhD in biophysics atKings College, London University, UK, followed by a postdocfellowship at the Biochemistry Department of Princeton Uni-versity, USA. In 1975 she became a group leader at the Max-Planck Institute for Molecular Genetics in Berlin, Germany,returning to the UK in 1980 to Bristol University. She iscurrently professor of biological chemistry and director ofthe Exeter Biocatalysis Centre, School of Biosciences. Shehas over 110 refereed publications to her name and haspresented her research work internationally. Her currentresearch involves the structural andmechanistic characteriza-tion of human enzymes involved in oxidative stress, aggrega-tion of b-amyloid protein in Alzheimers disease, GSTs andresponse to anticancer drugs and mutations in key enzymesresulting in diabetes and fructose intolerance.Jennifer A. Littlechilds, Universities of Exeter and Ply-

mouth, PeninsulaMedical School, St. Lukes Campus, Magda-len Road, Exeter EX1 2LU, UK

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Philip K.Moore received his BSc and PhD from the Universityof London. In recent years he was head of the Department ofPharmacology at the National University of Singapore and isnow professor of integrative pharmacology at Kings Collegein London. He has been researching biologically active gasesfor the last 30 years and in that time has reported the char-acterization of several inhibitors of nitric oxide synthase andlatterly hydrogen sulfide donor drugs. His major researchinterests are in the local regulation of blood flow and perme-ability, inflammatory mechanisms in arthritis and shock andin the regulation of local mediator biosynthesis and activity.Philip K. Moore, Kings College London, Pharmaceutical

Science Division, Franklin-Wilkins Building, 150 StamfordStreet, London SE1 9NH, UK

Michael P. Murphy received his BA in chemistry fromTrinity College, Dublin in 1984 and his PhD in biochemistryfrom Cambridge University in 1987. After spells in theUSA, Zimbabwe and Ireland, he took up a position in theBiochemistry Department for the University of Otago,Dunedin, New Zealand in 1992. In 2001 he moved to theMRC Dunn Human Nutrition in Cambridge, UK, where heis a group leader. Currently his special interests are intargeting small molecules such as antioxidants to mitochon-dria, and in understanding how modifications to the thiolstatus of mitochondrial proteins contributes to oxidativedamage and redox signaling.Michael P. Murphy, Medical Research Council Dunn

Human Nutrition Unit, Wellcome Trust/Medical ResearchCouncil Building, Hills Road, Cambridge CB2 0XY, UK

Following a BSc in human nutrition and dietitics at theUniversity of Dublin, Valerie ODonnell gained her PhD inbiochemistry from the University of Bristol, in the field ofneutrophil NADPH oxidase. This led to two postdoc posi-tions, first at the University of Bern, Switzerland, and then theUniversity of Alabama at Birmingham, USA. In Switzerland,she worked on mechanisms of TNF cytotoxicity and freeradical generation in fibroblasts. At UAB, she worked withBruce Freeman on interactions of nitric oxide with oxidizinglipids. Following this, she returned to the UK on a WellcomeTrust RCD Fellowship at Cardiff University. She is currently agroup leader, working in the area of free radical and lipidbiochemistry, related to vascular inflammation.

XXVIII List of Authors