Book Review
Stressful Oxygen Measurements.
methods suitable for the measurement of oxidation and antioxidants by luminescence; A Review of ''Free Radical and Antioxidant Protocols,'' Donald Armstrong (ed), Methods in Molecu-and the use of the computerized, versatile kinetic microplate reader, which can be ap-lar Biology Vol. 108, 1998. ISBN 0-89603-472-0. Totowa, NJ: Humana Press.
plied to many assays (6 protocols). The latter procedure is superior to conventional spectrophotometry in that it enables the si-The interest in reactive oxygen species and free radimultaneous measurement of discontinuous cals in biological systems has a long history and has and continuous kinetics of oxidation of been increasing exponentially over the past three demany samples. The use of small quantities cades. Oxidative stress is exerted by singlet oxygen, of reagents brings about considerable savsuperoxide, hydrogen peroxide, and hydroxyl radicals.
ing. The resulting lipid peroxides, lipid hydroperoxides, and
The organization of this book is methodological and carbon-or nitrogen-centered radicals have been shown didactic. It is based on the type of oxygen radical, to cause a plethora of oxidative damages to cellular damage, or antioxidant activity being measured rather proteins, membranes, and nucleic acids. Oxidative than on the methodology used. The reader is first predamage has also been associated with a variety of dissented with classical and modern methods for measureorders and pathologies, including cancer and aging. ment of oxygen consumption, followed by up-to-date This book is a timely, comprehensive collection of physical methods for the detection of free radicals (low general and specific methodologies for the detection frequency EPR spectrometry and single photon countand measurement of reactive oxygen species and their ing). There are several protocols for the fluorimetric, biological products, and for the study of antioxidant spectrophotometric, and luminometric determination of activities.
radicals in various body fluids and tissues, and for The book is in three parts: techniques for the meathe determination of oxidative damage to lipoproteins, surement of oxidative stress, techniques for the meafatty acyl esters, steroids, creatinine, and -everysurement of antioxidant activities, and techniques apbody's favorite molecule -DNA. plicable to both purposes. Each chapter consists of a Especially useful is the relatively simple procedure single protocol and is neatly and consistently organized for the elimination of thiobarbituric acid (TBA)-reacto contain a Theoretical Introduction, Materials, Instrutive glucose and water-soluble aldehydes, and for the mentation, Reagents, Methods, Results, Evaluation of inhibition of nonspecific reactions of TBA with subthe Results, and References. Some chapters contain stances such as sialic acid. As a result, the amounts of recommendations for the method of choice and for appropriate working conditions. Notes of interest and lipid peroxides present can be accurately expressed in terms of molar concentrations of malondialdehyde points of caution are often included as well.
The forty protocols included here address the follow-(MDA) in lieu of the popular, but less specific, ''TBAreactive material.'' An important addition is the new ing areas:
Part I: EPR, fluorescence, and chemiluminescence protocol for the immunodetection of MDA with antibodies against MDA-modified proteins; this allows di-methods for the measurement of reactive oxygen species, lipid peroxides and hydroper-rect measurement in body fluids without the need for separation and extraction methods, and could possibly oxides, malondialdehyde, and for oxidized cellular constituents in various body fluids be modified for the convenient detection of oxidative damage in tissue sections. and tissues (25 protocols). Part II: Measurement of the concentrations of antiox-
The protocols are concise and clearly presented. The introductions to theory are short, leading directly to the idants, such as vitamins E, C, and coenzyme Q10, and nonvitamin antioxidants such as practical items. This makes the book especially useful for the trained researcher who is more familiar with lipoic acid and glutathione (9 protocols). Part III: Novel methods for the generation of radicals the theoretical background. The protocols that describe more recent methods and require sophisticated equip-and radical scavenging systems with photo-Fenton reagent and caged compounds; ment are appropriately more detailed. A very useful