Diagnosis of OS and ROS Levels

The chemiluminescence assay is a direct method of quantifying extracellular ROS levels. This assay uses lucigenin and luminol to assess the generation of the superoxide anion and hydrogen peroxide [76]. Lucigenin detects superoxide anion, while luminol detects hydrogen peroxide. These agents are oxidizable substrates that react with certain oxygen species in order to measure levels of ROS. The chemilumines-cence assay or calorimetric assay can also be used to measure the total antioxidant

Fig. 24.1 The general sources, mechanisms, and consequences of oxidative stress (OS) on male fertility are summarized. Clinical conditions related to OS include idiopathic infertility, leukocy-tospermia, varicocele, genitourinary tract infection, environmental and lifestyle factors. OS acts through several mechanisms which lead to subfertility, such as lipid peroxidation, DNA damage, and apoptosis. OS can lead to several consequences related to male fertility, both in an in vivo and in vitro setting

Fig. 24.1 The general sources, mechanisms, and consequences of oxidative stress (OS) on male fertility are summarized. Clinical conditions related to OS include idiopathic infertility, leukocy-tospermia, varicocele, genitourinary tract infection, environmental and lifestyle factors. OS acts through several mechanisms which lead to subfertility, such as lipid peroxidation, DNA damage, and apoptosis. OS can lead to several consequences related to male fertility, both in an in vivo and in vitro setting capacity (TAC) of a semen sample. Subsequently, a ROS-TAC score is used to quantify if an imbalance exists between ROS and free radical scavengers. Low ROS-TAC scores are indicative of overall OS and have been identified in patients with idiopathic infertility, varicocele, and male accessory gland infection [77, 78]. These scores may be useful in predicting infertility when compared to using just ROS or TAC scores [78]. Patients with varicocele and other conditions associated with male factor infertility have low TAC scores [77], indicating an inability to scavenge ROS.

An indirect method of testing for ROS levels is through the examination of malondialdehyde (MDA) levels. MDA is a by-product of lipid peroxidation, which can be measured to detect the amount of lipid peroxidation in a semen sample. The MDA levels correlate with sperm motility and the potential for sperm-oocyte fusion [79]. This assay is useful in determining ROS levels before ART procedures and in diagnosing patients with subfertility [80]. Some studies have shown that high MDA levels correlate with other decreased sperm parameters such as concentration and motility [81, 82]. Higher MDA levels have been found in patients with varicocele when compared to controls, indicating increased peroxidative activity [31].

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