Methods In Biological Oxidative Stress

Oxidative damage appears to play a central role in the development of a wide range of tissue pathology, including neurodegenerative disease, drug side-effects, xenobiotic toxicity, carcinogenesis, and the aging process, to name just a few. Because of the centrality of oxidative processes to normal and abnormal tissue function, it has become imperative to develop appropriate analytical techniques to facilitate the quantitation of significant reactants. Without advances in methodology, corresponding advances in our knowledge of underlying biochemical events will be necessarily limited. Drs. Hensley and Floyd have done an outstanding job of assembling the work of world-class experts into Methods in Biological Oxidative Stress. The contributors have presented concise, yet thorough, descriptions of the state-of-the-art methods that any investigator working in the field needs to access. Mannfred A. Hollinger v Preface Free radicals and reactive oxidizing agents were once ignored as biochemical entities not worth close scrutiny, but are now recognized as causes or contributing factors in dozens, if not hundreds, of disease states. In addition, free radical metabolisms of xenobiotics have become increasingly important to pharmacologists. Accordingly, the need has arisen to accurately quantify reactive oxygen species and their byproducts. Methods in Biological Oxidative Stress is practical in scope, providing the details of up-to-date techniques for measuring oxidative stress and detecting oxidizing agents both in vitro and in vivo. The contributors are recognized experts in the field of oxidative stress who have developed novel strategies for studying biological oxidations.

This book discusses the importance of oxidative stress, related biomarkers, and its diagnostic methods including bio-analytical and advanced detection systems. Oxidative stress is associated with diminished capacity of a biological system to overcome the overproduction of reactive oxygen species. Since oxidative stress has been suggested as a causative factor in many diseases, its prevention is vital, and there is an urgent need for oxidative stress monitoring using in vitro and in vivo models. Interdisciplinary approaches have lead to the development of various oxidative stress monitoring models for real-world clinical and biomedical applications. The development of such methods requires a broad understanding of biology, chemistry, molecular biology, immunology and microbiology. As such, the book is a valuable resource for students, researchers and clinicians interested in the fundamental as well as applied aspects of oxidative stress associated diseases.

Expanding upon the research elucidated by the first volume of this collection, Advanced Protocols in Oxidative Stress II presents thirty additional cutting-edge chapters focusing on novel techniques for detecting ROS/RNS, unique AOX technology and applications, gene expression and biostatistics for evaluating OS-derived experimental data. The international panel of authors also provide animal models and numerous studies concentrating on mitochondria during hypoxic conditions using advanced methods for pO2, peroxynitrate, reactive S-nitrosothiols, lipid peroxides, COX, and the mitochondrial membrane potential. Due to the dynamic nature of this topic, this book is the second of several volumes of Advanced Protocols in Oxidative Stress, all included in the highly successful Methods in Molecular BiologyTM series. As part of the series, the chapters of this volume present brief introductions to the respective subjects, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and tips on troubleshooting to ensure easy replication of the technology involved. Authoritative and convenient, Advanced Protocols in Oxidative Stress II is an ideal desk reference for scientists wishing to further the research in this exciting, unique, and vital field of study.

This book describes the methods of analysis and determination of oxidants and oxidative stress in biological systems. Reviews and protocols on select methods of analysis of ROS, RNS, oxygen, redox status, and oxidative stress in biological systems are described in detail. It is an essential resource for both novices and experts in the field of oxidant and oxidative stress biology.

The first protocols book, Free Radical and Antioxidant Protocols (1) was published in late 1998. Sections were divided into three parts, covering selected biochemical techniques for measuring oxidative stress, antioxidant (AOX) activity, and combined applications. In choosing the 40 methods to be included in that book, I realized there were considerably more of equal value than that which we could have presented in a single volume. To produce a comprehensive resource, this book and a third are being compiled to expand coverage of the field. A summary of papers (2) published on this important subject emphasizes the continuing rapid growth in oxidative stress investigations relating to our understanding of biochemical reactions, their relevance to pathophysiological mechanisms, how disease may arise, and how therapeutic intervention may be achieved(3). Although there is some overlap between the categories, the ana- sis shown below illustrates where current studies are concentrated and are almost evenly distributed between free radicals and AOX. Over the last 4 yr, there has been a 55% increase in the number of papers published in the area.

Protocols books specializing in measuring free radical and antioxidant biomarkers began to be published in 1998. Many of these methods are currently finding use in diagnostic medicine. Advanced Protocols in Oxidative Stress I covers the field of oxidative stress with state-of-the-art technology to utilize in research, contributed by an international panel of experts renowned for developing new procedures and methods. Included are sections on reactive oxygen and nitrogen species techniques, antioxidant technology and application, methods for analyzing gene expression, the exciting new area of oxidative stress and stem cell differentiation and specific biostatistical evaluation of biomarkers. This volume presents the current high-tech methodologies and provides a perspective on the diversity of applications in the ever-emerging field of free radical reactions and antioxidants. Due to the dynamic nature of this topic, this book will be the first of several volumes of Advanced Protocols in Oxidative Stress, all part of the highly successful Methods in Molecular BiologyTM series. As part of the series, the chapters include a brief introduction to the material, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and tips on troubleshooting and ensuring replication of technology. Cutting-edge and convenient, Advanced Protocols in Oxidative Stress I is an ideal desk reference for scientists wishing to further this research in this exciting, unique and vital field of study.

Advanced Protocols in Oxidative Stress I covers the field of oxidative stress with state-of-the-art technology to utilize in research, contributed by an international panel of experts renowned for developing new procedures and methods.

In our first protocols book, Free Radical and Antioxidant Protocols (1), r- erence to in vivo, ex vivo, or in situ techniques were few compared to classical biochemical assays and only 6 of the 40 chapters were concerned with these applications. In our second book, Oxidative Stress Biomarkers and Antioxidant Protocols (2), which is being published concurrently with this third volume, Oxidants and Antioxidants: Ultrastructure and Molecular Biology Protocols, the number of such chapters has increased. The literature dealing with histoche- cal/cytochemical and immunohistochemical techniques and staining to identify cellular/subcellular sites of oxidative stress has expanded rapidly, as has the molecular biology methodology used to analyze free radical and antioxidant (AOX) reactions, as well as the monitoring of living tissue. A two-way search was performed for each technique listed in Table 1, coupled with “oxidative stress” using the PUBMED search engine from the National Library of Medicine at NIH. Most of the techniques involved in m- suring oxidative stress employ molecular biology or ultrastructural approaches. Of these techniques, histology, polymerase chain reaction, and Western blotting are the most widely used. Several forms of therapy are now available for patients with increased oxidative stress. In addition to standard antioxidant therapy supplementation in vivo and in vitro, photodynamic therapy (PDT) employs excitation of a photon-emitting compound delivered systemically for free radical-mediated necrosis of affected tissues, and stem cells are also being used to induce signaling events or replace antioxidant enzymes.