Free Radicals And Antioxidant Protocols

This cutting-edge and updated book offers methods for the rapid detection of RONs and redox stress. It includes in-depth analysis of natural and synthetic antioxidants, and also of DNA oxidation, oxidative lipidomics, and biomarkers.

This volume presents an outstanding collection of state-of-the-art methodologies for quantifying free radical and antioxidant analytes in tissue and body fluids using experimental models and in vitro procedures. These user-friendly and easily reproducible techniques cover the essential tasks, including radical generating systems, direct measurement or trapping of reactive radical species and acute-phase proteins, and measurement of metabolic intermediates derived from the oxidation of lipids, proteins, and nucleic acids. There are also methods for the determination of vitamin, enzymatic, and water-soluble antioxidants, as well as of essential micronutrients and cofactors. The techniques take advantage of new instrumentation and technology-probes, photon counting, chemiluminescence, and caged compounds, with an emphasis on HPLC-and are adaptable to a wide range of applications. The laboratory-tested free-radical assays described here in detail will illuminate the study of both primary and secondary oxidative stress and contribute significantly to our understanding of the many disorders associated with this process.

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.

This volume presents an outstanding collection of state-of-the-art methodologies for quantifying free radical and antioxidant analytes in tissue and body fluids using experimental models and in vitro procedures. These user-friendly and easily reproducible techniques cover the essential tasks, including radical generating systems, direct measurement or trapping of reactive radical species and acute-phase proteins, and measurement of metabolic intermediates derived from the oxidation of lipids, proteins, and nucleic acids. There are also methods for the determination of vitamin, enzymatic, and water-soluble antioxidants, as well as of essential micronutrients and cofactors. The techniques take advantage of new instrumentation and technology-probes, photon counting, chemiluminescence, and caged compounds, with an emphasis on HPLC-and are adaptable to a wide range of applications. The laboratory-tested free-radical assays described here in detail will illuminate the study of both primary and secondary oxidative stress and contribute significantly to our understanding of the many disorders associated with this process.

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.

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.

Presents information from significant peer-reviewed studies showing that increased oxidative stress has an important role in numerous medical disorders. Describes current concepts of life expectancy, the aging process, and the presence of free radicals in biological systems. Explores the relationship between free radical and increased oxidative stress in medical disorders, including atherosclerosis, ischemia/reperfusion injury, pulmonary diseases, liver and kidney disorders, cancer, and neurodegenerative disorders.

Free Radicals in Biology and Medicine has become a classic text in the field of free radical and antioxidant research. Now in its fifth edition, the book has been comprehensively rewritten and updated whilst maintaining the clarity of its predecessors. Two new chapters discuss 'in vivo' and 'dietary' antioxidants, the first emphasising the role of peroxiredoxins and integrated defence mechanisms which allow useful roles for ROS, and the second containing new information on the role of fruits, vegetables, and vitamins in health and disease. This new edition also contains expanded coverage of the mechanisms of oxidative damage to lipids, DNA, and proteins (and the repair of such damage), and the roles played by reactive species in signal transduction, cell survival, death, human reproduction, defence mechanisms of animals and plants against pathogens, and other important biological events. The methodologies available to measure reactive species and oxidative damage (and their potential pitfalls) have been fully updated, as have the topics of phagocyte ROS production, NADPH oxidase enzymes, and toxicology. There is a detailed and critical evaluation of the role of free radicals and other reactive species in human diseases, especially cancer, cardiovascular, chronic inflammatory and neurodegenerative diseases. New aspects of ageing are discussed in the context of the free radical theory of ageing. This book is recommended as a comprehensive introduction to the field for students, educators, clinicians, and researchers. It will also be an invaluable companion to all those interested in the role of free radicals in the life and biomedical sciences.