Oxidants and Antioxidants

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.

Includes supplementary material: sn.pub/extras

Klappentext
It is now feasible to identify subcellular sites where oxidative stress reactions occur and to determine gene expression relevant to those complex pathways. In Oxidants and Antioxidants: Ultrastructure and Molecular Biology Protocols, a panel of world authorities present cutting-edge molecular and ultrastructure techniques for investigating the pathophysiology of oxidative stress. Described in step-by-step detail, these readily reproducible methods use the latest in immunodetection, X-ray diffraction, confocal microscopy, in situ hybridization, chemiluminescence, and cytofluorescence. Additional techniques take advantage of electron paramagnetic resonance, advanced glycation endproducts, Western blotting analysis, transcription factors, p47phox translocation, and microsphere delivery systems. The collection emphasizes the visual observation of free radicals or products altered by oxidative stress, and includes numerous in vivo, ex vivo, and in vitro applications. Two companion volumes, Oxidative Stress Biomarkers and Antioxidant Protocols (2002), and Free Radical and Antioxidant Protocols (1998), focus on the related biochemical and cellular biological methods.
Authoritative and highly practical, Oxidants and Antioxidants: Ultrastructure and Molecular Biology Protocols provides a comprehensive collection of powerful ultrastructure and molecular tools for the evaluation of metabolic disorders linked to oxidative stress and the aging process.

Inhalt
Techniques for Histological Identification.- Cytochemical Localization of H2O2 in Biological Tissues.- Localization of Intracellular Lipid Hydroperoxides Using The Tetramethylbenzidine Reaction for Transmission Electron Microscopy.- The Immunohistochemical Localization of Glutathione Peroxidase.- 4-Hydroxy-2-Nonenal (4-HNE) Staining by Anti-HNE Antibody.- Immunohistochemical Detection of Protein Oxidation.- Indirect Immunofluorescence Detection of Protein-Bound 4-Hydroxynonenal in Tissue Sections and Isolated Cells.- Techniques for Subcellular Localization.- Methods for Studying the Binding of Advanced Glycated Proteins to Receptors for Advanced Glycation Endproducts (AGE Receptors).- Measurement of Blue Fluorescence as a Protein Marker for Oxidized Membranes.- X-Ray Diffraction Analysis of Membrane Structure Changes with Oxidative Stress.- Detection of Estrogen Receptor by In Situ Hybridization.- Tumor Necrosis Factor-? Quantification and Expression by In Situ Hybridization.- Techniques for Molecular Biology.- Translocation of p47phox and Activation of NADPH Oxidase in Mononuclear Cells.- Activation of Nuclear Factor-?B (NF-?B) in Mononuclear Cells (MNC).- Assay of Intracellular Hydrogen Peroxide Generation in Activated Cytometry Individual Neutrophils by Flow.- DNA-Binding Activity of Hypoxia-Inducible Factors (HIFs).- Analysis of Heat-Shock Transcription Factor and Element-Binding Activity.- Slot-Blot Hybridization in Studying Gene Expression During Oxygen Deprivation.- An Expression System for a Transporter of Iron and Other Metals.- Analysis of Gene Expression Following Oxidative Stress.- Measurement of Immunoglobulin G Oxidation by Western-Blot Analysis.- Thioredoxin and Redox Regulation of the Nuclear Receptor.- Analysis of Phospholipid HydroperoxideGlutathione Peroxidase mRNA.- Expression of Human Phospholipid Hydroperoxide Glutathione Peroxidase.- Techniques for In Vivo/Ex Vivo/In Vitro Applications.- Superoxide Production in the Islet of Langerhans Detected by the MCLA Chemiluminescence Method.- In Vivo Detection of Transition Metals and Nitrosyl-Heme Complexes Using Ex Vivo Electron Paramagnetic Resonance Spectroscopy.- Spectral-Spatial Electron Paramagnetic Resonance Imaging (EPRI) in Skin Biopsies at 9.5 GHz.- In Vivo In Situ Detection of Nitric Oxide Using Low-Frequency EPR Spectroscopy.- of NOS II Gene into Primary Cultures of Bovine and Human Endothelial Cells.- Detection of Peroxynitrite-Induced Protein and DNA Modifications.- Immunochemical Detection of a Fluorophore Derived from the Lipid Peroxidation Product 4-Hydroxy-2-Nonenal and Lysine.- Liver Slice Technology as an In Vitro Model for Metabolic and Toxicity Studies.- Cytofluorescence Techniques for the Visualization of Distinct Pools of Protein Thiols at the Single Cell Level.- Preparation of Microspheres and Incorporation of Lipid Hydroperoxide for Sustained Release Studies.- Measurement of Carotenoids in the Living Primate Eye Using Resonance Raman Spectroscopy.- Interstitial Photodynamic Therapy with Moving Exposure Fiber.- Statistical Analysis.