Measuring ROS Production

While detecting the presence of ROS-producing cells is possible, the direct measurement of in vivo ROS concentrations is challenging due to the fact that ROS are very short-lived [1]. In the field of male reproductive studies, the measurement of OS is quantified only indirectly by the presence of ROS. The quantification of ROS has been promoted as a valuable test to be performed in the assessment of seminal plasma that is to be used for ART as it may be beneficial in predicting end points such as the fertilization rate [5]. The detection and quantification of ROS as a means of studying the role of OS is carried out in varying ways.

One of these methods is the luminol-dependant chemiluminescence assay, which indirectly measures ROS produced by spermatozoa by means of a sensitive probe, which can either be luminol or lucigen [31, 41]. The principle of this test is based on the concentration of released luminescence in a luminometer [1, 72]. This is the amount of light that is generated with the interaction between the ROS present in the sample and a specific chemiluminescent probe [41]. This particular assay is a reproducible test with high sensitivity and specificity and allows for both the extra- and intracellular ROS to be measured [5, 31]. In conjunction with quantifying and identifying ROS in the semen sample by means of this particular assay, the relationship between the presence of leukocytospermia and the excessive ROS concentrations can be investigated. This can be done by determining leukocytospermia by performing the Endtz test, which is a myeloperoxidase staining technique based on the peroxidase activity of PMN [41]. An Endtz positive semen sample has been proven as an indicator of positive chemiluminescence for ROS [41].

The second test that can be used to determine if ROS is generated is by the measurement of lipid peroxidation through the Lipid Peroxidation Assay (LPO) [1]. Lipid peroxidation in spermatozoa is initiated by the presence of ROS and results in the generation of lipid hydroperoxides [1] which are stable molecules under normal physiological conditions [44]. However, under conditions of elevated ROS and resultant OS, malondialdehyde is produced from the decomposition of lipid peroxides [44]. The concentrations of this particular compound can be measured in biochemical assays to assess the level of peroxidative damage [73], An additional third test to assess the level of seminal OS is the ROS-TAC score. This particular method combines the concentrations of TAC and ROS in semen samples and is quantified by using a statistical formula that allows for the optimal quantification of seminal OS [44, 54] . Owing to cases where normozoospermic males suffer from infertility, which may be the result of seminal OS, the ROS-TAC score has been suggested as an advantageous means of assessing OS in comparison to ROS alone when distinguishing between fertile and infertile subjects [54]. Due to the clinical relevance of identifying OS due to its causative role in infertility, research has led to the development of an inexpensive assay: the photometric nitro blue tetrazolium (NBT) assay which allows for the measurement of seminal ROS production [74] . This finding serves as a hopeful indicator of what future research into OS may hold.

Pregnancy Guide

Pregnancy Guide

A Beginner's Guide to Healthy Pregnancy. If you suspect, or know, that you are pregnant, we ho pe you have already visited your doctor. Presuming that you have confirmed your suspicions and that this is your first child, or that you wish to take better care of yourself d uring pregnancy than you did during your other pregnancies; you have come to the right place.

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