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Intended for molecular and cellular biologists, this volume contains 33 papers outlining reproducible techniques for evaluating the perturbations in cell function resulting from increased oxidative stress. It covers methods for oxidized lipids, proteins, carbohydrates, and nucleic acids, fat and water soluble vitamins, flavonoids, and metallothionein. It also details improved statistical interpretations, correcting for random measurement error. Charts, diagrams, and other images support the text.Book News, Inc.

Book Info

Univ. of Florida, Gainesville. Features reproducible techniques employing the latest technology; extensive coverage of techniques for oxidized lipids, proteins, carbohydrates, and nucleic acids; methods for fat and water soluble vitamins, flavonoids, and metallothionein; and improved statistical interpretation.

Preface
Contributors
Pt. I Techniques for Free Radical Derived Biomarkers
1 Human Xanthine Oxidoreductase Determination by a Competitive ELISA 3
2 Simultaneous Determination of Polyunsaturated Fatty Acids and Corresponding Monohydroperoxy and Monohydroxy Peroxidation Products by HPLC 13
3 Determination of Products of Lipid Oxidation by Infrared Spectroscopy 21
4 Detection of Docosahexaenoic Acid Hydroperoxides in Retina by Gas Chromatography/Mass Spectrometry 29
5 Detection of Lipid Hydroperoxide-Derived Protein Modification with Polyclonal Antibodies 37
6 Techniques for Determining the Metabolic Pathways of Eicosanoids and for Evaluating the Rate-Controlling Enzymes 45
7 Mass Spectrometric Quantification of F[subscript 2]-Isoprostanes as Indicators of Oxidant Stress 57
8 Formation of Apolipoprotein AI-AII Heterodimers by Oxidation of High-Density Lipoprotein 67
9 Detection of Certain Peroxynitrite-Induced DNA Modifications 77
10 Hydroxyl and 1-Hydroxyethyl Radical Detection by Spin Trapping and GC-MS 89
11 Analysis of Aliphatic Amino Acid Alcohols in Oxidized Proteins 101
12 Rapid Determination of Glutamate Using HPLC Technology 111
13 A Rapid Method for the Quantification of GSH and GSSG in Biological Samples 117
14 Protein Carbonyl Measurement by ELISA 123
15 N[actual symbol not reproducible]-(carboxymethyl)lysine (CML) as a Biomarker of Oxidative Stress in Long-Lived Tissue Proteins 129
16 Measurement of S-Glutathionated Hemoglobin in Human Erythrocytes by Isoelectric Focusing Electrophoresis 139
17 Oxidation of Cellular DNA Measured with the Comet Assay 147
18 Measurement of DNA Double-Strand Breaks with Giant DNA and High Molecular-Weight DNA Fragments by Pulsed-Field Gel Electrophoresis 161
19 Evaluation of Antibodies Against Oxygen Free Radical-Modified DNA by ELISA 171
Pt. II Techniques for Antioxidant Biomarkers
20 Simultaneous Analysis of Multiple Redox-Active Metabolites from Biological Matrices 185
21 Determination of Uric Acid in Urine by Fast-Scan Voltammetry (FSV) Using a Highly Activated Carbon Fiber Electrode 195
22 Measurement of [alpha]-Tocopherol Turnover in Plasma and in Lipoproteins Using Stable Isotopes and Gas Chromatography/Mass Spectrometry 209
23 Analysis of Tocotrienols in Different Sample Matrixes by HPLC 221
24 Measurement of [beta]-Carotene 15,15'-Dioxygenase Activity by Reverse-Phase HPLC 233
25 Ubiquinol/Ubiquinone Ratio as a Marker of Oxidative Stress 241
26 Catechol- and Pyrogallol-Type Flavonoids: Analysis of Tea Catechins in Plasma 247
27 Pyruvate Dehydrogenase Complex as a Marker of Mitochondrial Metabolism: Inhibition by 4-Hydroxy-2-Nonenal 255
28 Ceruloplasmin Detection by SDS-PAGE, Immunoblotting, and In Situ Oxidase Activity 265
29 Metallothionein Determination by Isocratic HPLC with Fluorescence Derivatization 273
30 Quantification of Oxidized Metallothionein by a Cd-Saturation Method 285
31 Fractionation of Herbal Medicine for Identifying Antioxidant Activity 293
32 Designing Safer (Soft) Drugs by Avoiding the Formation of Toxic and Oxidative Metabolites 301
33 Statistical Correction of the Area Under the ROC Curve in the Presence of Random Measurement Error and Applications to Biomarkers of Oxidative Stress 313
Index 319