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To better understand how researchers make conclusions about the necessity and effectiveness of antioxidant supplements, knowledge of their measurement techniques is required. Basically, three methods are common to assessing the effectiveness of antioxidant compounds in athletes: measurement of the oxidized products or molecules, measurement of tissue damage, and ultimately, measurement of performance itself. Investigators can sample serum, urine, breath, or tissue specimens for oxidized compounds generated as a result of free radical activity. These products include malondialdehyde (MDA), thiobarbituric acid-reactive substances (TBARS), pentane and ethane, conjugated dienes, and lipid hydroperoxides. Of these, high-performance liquid chromatography (HPLC) measurement of lipid hydroperoxides is the most direct assessment of oxidative damage. Because of inconsistencies among these techniques and the variability of oxidized products, the best research takes into account more than one of the oxidation products and may investigate more than one medium (i.e., breath and serum).
In addition to indices of oxidative damage, physiologists interested in muscle damage and recovery also measured damage indirectly through cellular damage markers. Since lipid membrane peroxidation leads to abnormalities in the cell membrane, intracellular enzymes may leak from the cell and into the blood plasma with antioxidant damage. These enzymes can be measured in the blood as indirect markers of cell membrane damage. The most frequently measured enzymatic markers of muscle damage in the serum are as creatine kinase (CK), alanine aminotransferase (ALT-formerly known as SGPT), aspartate aminotransferase (AST -formerly SGOT), and lactate dehydrogenase (LDH). Muscle proteins can also be catabolized as a result of mechanical and oxidative damage, therefore, urinary nitrogen excretion can also be quantified. Taking the nitrogen excretion concept further, nitrogen balance (NBAL) studies and even stable isotope methodology can be used to assess muscle damage via protein breakdown.
Perhaps most pertinent to an athlete's bottom line are a variety of exercise performance tests that can be used. These range from treadmill and cycle protocols to strength measurements. Such tests can assess both ergogenic benefits from [antioxidant] supplementation and recovery rate after strenuous bouts of exercise. In essence, the practical effects of antioxidant supplementation can be assessed by the determination of whether performance can be improved or whether a high level of performance can be maintained with repeated bouts of exercise.
When considering the data obtained from the a forementioned measures, often findings are inconsistent. Much of the inconsistent findings in exercise and free radical/antioxidant research may be attributed to the lack of uniformity in study design regarding protocol, especially for performance testing. When evaluating this research, it is important to note the type, intensity, and duration of exercise used as well as the duration of the recovery period between multiple bouts. Since performance testing tends to yield the most inconsistent results in antioxidant research, some authors have criticized the use of performance measures. Physical performance is a multifactorial process, and therefore performance testing may not serve as a good indicator of the effects of antioxidant supplementation. Although exercise testing may not be an extremely sensitive measure, numerous studies have shown positive effects of antioxidant supplementation and therefore, if protocol uniformity is attempted, perhaps the data would show more consistent results in a specific direction.
In addition to protocol concerns, populationspecific variables are important to consider. When evaluating the results from antioxidant research, it is important to note the type of population that the subjects were drawn from (i.e., weight lifters, cyclists, runners, old, young, male, female, etc.). Of course, animal ex vivo (pre-treated human cells examined outside the body) or in vitro (in a test tube or dish) data are sometimes the only available source of information. In such cases, recognize that humans may react differently but are generally considered similar until proven otherwise.
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