Free Radical and Antioxidant Protocols

About this book

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.

Contents

Part I.Techniques for the Measurement of Oxidative Stress. Oxygen Consumption Methods: Xanthine Oxidase and Lipoxygenase Istvan Stadler. Spin Trapping and Electron Paramagnetic Resonance Spectroscopy Daniel J. Brackett Gemma Wallis Michael F. Wilson and Paul B. McCay. 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. Single Photon Counting Yoshinori Mizuguchi. Measurement of Reactive Oxygen Species in Whole Blood and Mononuclear Cells Using Chemiluminescence Kuldip Thusu Ehad Abel-Rahman and Paresh Dandona. Assay of Phospholipid Hydroperoxides by Chemiluminescence-Based High-Performance Liquid ChromatographyYorihiro Yamamoto Yasuhiro Kambayashi and Takako Ueda. Sensitive and Specific Fluorescent Probing of Oxidative Stress in Different Classes of Membarne Phospholipids in Live Cells Using Metabolically Integrated cis-Parinaric AcidValerian E. Kagan Vladimir B. Ritov Yulia Y. Tyurina and Vladimir A. Tyurin. Aromatic Hydrozylation: Salicylic Acid as a Probe for Measuring Hydroxyl Radical Production Andrea Ghiselli. Simple Assay for the Level of Total Peroxides in Serum or Plasma Kunio Yagi. Simple Procedure for Specific Assay of Lipid Hydroperoxides in Serum or Plasma Kunio Yagi. Antibodies Against Malondialdehyde-Modified Proteins: Induction and ELISA Measurements of Specific Antibodies Yves Chancerelle J. Mathieu and J. F. Kergonou. Oxidized LDL and Lp(a): Preparation Modification and Analysis Jan Galle and Christoph Wanner. Oxidized and Unoxidized Fatty Acyl Esters Odile Sergent and Josiane Cillard. Synthesis of Lipid and Cholesterol Hydroperoxide Standards Donald Armstrong and Richard W. Browne. Separation of Hydroxy and Hydroperoxy Polyunsaturated Fatty Acids by High-Pressure Liquid Chromatography Richard W. Browne and Donald Armstrong. Products of Creatinine with Hydroxyl Radical as a Useful Marker of Oxidative Stress In Vivo Kazumasa Aoyagi Sohji Nagase Akio Koyama Mitsuharu Natitam and Shizuo Tojo. Quantitative Analysis of Peptide and Protein Changes in Ischemic Hippocampal Tissue by HPLCThomas M. Wengenack J. Randall Slemmon and J. Mark Ordy. Electrochemical Detection of 8-Hydroxy -2- Deoxyguanosine Levels in Cellular DNA Thomas M. Nicotera and Sofia Bardin. Nitric Oxide Synthase Ahmad Aljada and Paresh Dandona. Quantitation of Heme Oxygenase (HO-1) Copies in Human Tissues by Competitive RT/PCR Nader G. Abraham. A Fluorescent-Based Assay for Measurement of Phospholipase A2 ActivityThomas M. Nicotera and Glenn Spehar. Autofluorescent Ceroid/Lipofuscin Dazhong Yin and Ulf Brunk. In Vivo Technique for Autofluorescent Lipopigments Fran+ois C. Delori and C. Kathleen Dorey. An Ex Vivo Erythrocyte Model for Investigating Oxidative Metabolism Luigia Rossi and Mauro Magnani. Purification of Vesicular Carriers from Rat Hepatocytes by Magnetic Immunoadsorbtion Lucian Saucan. Part II.Techniques for the Measurement of Antioxidant Activity. Simultaneous Determination of Serum Retinol Tocopherols and Carotenoids by HPLC Richard W. Browne and Donald Armstrong. EPR Measurements of Nitric Oxide-Induced Chromanoxyl Radicals of Vitamin E: Interactions with Vitamin C Valerian E. Kagan and Nikolai V. Gorbunov. Nonvitamin Plasma Antioxidants Nicholas J. Miller. Regulatory Antioxidant Enzymes C. E. Pippenger Richard W. Browne and Donald Armstrong. In Vitro Screening for Antioxidant Activity Donald Armstrong Tadahisa Hiramitsu and Takako Ueda. Antioxidant Activity of Low-Density Lipoprotein Nicholas J. Miller and George Paganga. Lipoic Acid as an Antioxidant: The Role of Dihydrolipoamide Dehydrogenase Mulchand S. Patel and Young Soo Hong. Reduced Glutathione and Glutathione Disulfide Richard W. Browne and Donald Armstrong. Analysis of Coenzyme Q10 Content in Human Plasma and Other Biological Samples Scott Graves Marianna Sikorska Henryk Borowy-Borowski Rodney J. H. Ho Tot Bui and Clive Woodhouse. Part III. Techniques Applicable to Both Oxidative Stress and Antioxidant Activity. A Simple Luminescence Method for Detecting Lipid Peroxidation and Antioxidant Activity In Vitro Minoru Nakano Takashi Ito and Tadahisa Hiramitsu. Analysis of Oxidation and Antioxidants Using Microtiter Plates Germ n Camejo Boel Wallin and Mervi Enoj"rvi. Trace Element Analysis of Biological Samples by Analytical Atomic Spectroscopy Hiroki Haraguchi Eiji Fujimori and Kazumi Inagaki. Energy Dispersive X-Ray Microanalysis Don A. Samuelson. Pro-Oxidant and Antioxidant Assays of Estrogens Joachim G. Liehr and Deodutta Roy. Artificial Radical Generating and Scavenging Systems: Photo-Fenton Reagent and Caged Compounds Seiichi Matsugo and Ken Fujimori. Index.

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