Determination of Oxidative Stress in Seminal Plasma

In order to determine seminal oxidative stress, two different approaches can be chosen, the direct measurement of ROS by a chemiluminescent method using luminol [8, 164] or other chemiluminescent substances, or the indirect assessment of oxidative stress by means of the measurement of MDA [165], which is a stable end product of LPO [107] [ Luminescent methods are fast and highly sensitive as light can be detected at very low emission rates. However, specialized equipment (luminometer) is necessary and not every Andrology laboratory may be in possession thereof. In the indirect method, MDA, as thiobarbituric acid reactive substance (TBARS) is measured by means of standard photometry, which is available in almost every laboratory. Thus, determination of TBARS is factually a measure of the degree of LPO, of which semen samples derived from infertile men were shown to have elevated levels [166], Furthermore, in abnormal semen samples TBARS levels correlate negatively with the sperm count [61], while in other studies the TBARS concentration was negatively correlated with the male germ cells' acrosin activity [66] as well as with fertilization rates in an IVF programme [62]. These data indicate that the determination of seminal TBARS levels can be indicative of male infertility. However, proper cut-off values have not been calculated yet.

For the direct measurement of ROS, the Agarwal group [167] has performed a receiver operating characteristics (ROC) analysis to establish a cut-off value of 0.145 x 106 cpm/20 x 106 spermatozoa to distinguish between fertile and infertile men with a sensitivity and specificity of the test of 87 and 57%, respectively. Although outputs from different luminometers cannot be directly compared because of different response characteristics of the photomultipliers, Henkel et al. (unpublished data) came to a similar result of 0.229 x 103 cpm/20 x 106 spermatozoa after ROC analysis for fertilization.

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