Free Radicals and Antioxidant Protocols

In recent years there has been a growing demand to achieve more rapid and accurate detection of RONs (reactive oxygen and nitrogen species), which affect normal physiological processes and pathological conditions, and also of redox stress. In Free Radicals and Antioxidant Protocols, Second Edition, world-renowned researchers respond to this need, considering the most recent analytical and technological aspects of the detection and determination of RONs, and of oxidative, nitrative, nitrosative, and redox stress in biological systems in vivo and in vitro. Chapters include protocols to establish biomarkers for the oxidative stress-induced genetic damage in cellular and animal models, provide novel methods and technologies for analyzing peroxidized lipids, describe techniques for gene therapy, and present methods of EPR imaging of free radicals and oxygen in vivo. Composed in the highly successful Methods in Molecular Biology'„¢ series format, each chapter contains a brief introduction, step-by-step methods, a list of necessary materials, and a Notes section which shares tips on troubleshooting and avoiding known pitfalls.Comprehensive and user-friendly, Free Radicals and Antioxidant Protocols, Second Edition, is an essential guide for the novice and expert researcher alike.

Preface	p. v
Contributors	p. xi
Reactive Oxygen and Nitrogen Species	p. 1
In Vivo Imaging of Free Radicals and Oxygen	p. 3
In Vivo Measurement of Tissue Oxygen Using Electron Paramagnetic Resonance Spectroscopy with Oxygen-Sensitive Paramagnetic Particle, Lithium Phthalocyanine	p. 29
Measurement of Plasma Nitrite by Chemiluminescence	p. 41
Determination of Glutathione, Mitochondrial Transmembrane Potential, and Cytotoxicity in H9c2 Cardiomyobiasts Exposed to Reactive Oxygen and Nitrogen Species	p. 51
Natural and Synthetic Antioxidants	p. 63
Phenolic Acids and Flavonoids: Occurrence and Analytical Methods	p. 65
Design, Synthesis, and Action of Antiatherogenic Antioxidants	p. 91
Preparation of Drug-Loaded Polymeric Nanoparticles and Evaluation of the Antioxidant Activity Against Lipid Peroxidation	p. 109
Nanoparticle and Iron Chelators as a Potential Novel Alzheimer Therapy	p. 123
Synthesis and Characterization of Polymer Nanocarriers for the Targeted Delivery of Therapeutic Enzymes	p. 145
Assessment of Antioxidant Activity of Eugenol In Vitro and In Vivo	p. 165
Cellular Oxidative Stress	p. 181
The Generation of Stable Oxidative Stress-Resistant Phenotypes in Chinese Hamster Fibroblasts Chronically Exposed to Hydrogen Peroxide or Hyperoxia	p. 183
A Simple Method for Effective and Safe Removal of Membrane Cholesterol from Lipid Rafts in Vascular Endothelial Cells: Implications in Oxidant-Mediated Lipid Signaling	p. 201
Superoxide Dismutase - A Target for Gene Therapeutic Approach to Reduce Oxidative Stress in Erectile Dysfunction	p. 213
Assessing the Reductive Capacity of Cells by Measuring the Recycling of Ascorbic and Lipoic Acids	p. 229
Biomarkers of Oxidative Stress: Methods and Measures of Oxidative DNA Damage (COMET Assay) and Telomere Shortening	p. 245
Simultaneous Analysis of Expression of Multiple Redox-Sensitive and Apoptotic Genes in Hypothalamic Neurons Exposed to Cholesterol Secoaldehyde	p. 263
Redox Homeostasis and Cellular Stress Response in Aging and Neurodegeneration	p. 285
Gene Therapy Techniques for the Delivery of Endothelial Nitric Oxide Synthase to the Lung for Pulmonary Hypertension	p. 309
DNA Oxidation, Oxidative Lipidomics, and Biomarkers	p. 323
A General Method for Quantifying Sequence Effects on Nucleobase Oxidation in DNA	p. 325
Analysis of Urinary 8-oxo-7,8-dihydro-2′-deoxyguanosine by Liquid Chromatography-Tandem Mass Spectrometry	p. 341
Oxidative Lipidomics of Apoptosis: Quantitative Assessment of Phospholipid Hydroperoxides in Cells and Tissues	p. 353
Simultaneous Analysis of Multiple Lipid Oxidation Products In Vivo by Liquid Chromatographic-Mass Spectrometry (LC-MS)	p. 375
Lipoxygenase-Catalyzed Phospholipid Peroxidation: Preparation, Purification, and Characterization of Phosphatidylinositol Peroxides	p. 387
Oxidized Low-Density Lipoprotein	p. 403
Detection and Localization of Markers of Oxidative Stress by In Situ Methods: Application in the Study of Alzheimer Disease	p. 419
Enzyme Immunoassay of Isoprostanes	p. 435
Application of Membrane Extraction with Sorbent Interface for Breath Analysis	p. 451
Subject Index	p. 469
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Free Radicals and Antioxidant Protocols

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