ROS and male subfertility

Excess ROS  may be a cause of your IUI failure: ROS are usually washed away with semen preparation in IUI /ART: Reactive oxygen species are metabolites of oxygen and include superoxide anion, hydrogen peroxide, hydroxyl radical and nitric oxide. ROS is produced by both spermatozoa and leucocytes. Increased number of sperms would produce more ROS. When present in excess they can initiate oxidative damage to cellular lipids, proteins and DNA. Most cells are equipped with enzymatic or non enzymatic antioxidant systems. The seminal plasma also has these enzymes. Now coming to sperm prepn , while preparing sperms, seminal plasma is removed and the sperms are rendered vulnerable to oxidative attack affecting sperm function. Tests can differentiate ROS produced from sperms and those from leucocytes. Chemiluminescent procedures employing probes such as luminal or lucigenin may be used to measure ROS production.

What is peroxidase staining?? Ans: The ejaculate contains cells, other than spermatozoa, referred to as round cells. These include germinal line cells sloughed cells from seminiferous tubules, epithelial cells from genitourinary tract, prostate cells, spermatogenic cells and leukocytes. A normal ejaculate should not contain more than 1 x 106 round cells/ml. The number of leucocytes should not exceed 1 x 106. The concentration of granulocytes in semen can be determined using peroxidase staining and hemocytometer counts. Round cells that lack polymorphonuclear morphology may he immature germinal cells (spermatocytes, spermatids) exfoliated epithelial cells. These may be degenerating and difficult to identify. To identify them special staining procedures with monoclonal antibodies are used. If round cells are more than 1 million it should be ascertained whether they are leucocytes or spermatids, as leucocytes more than 1 million indicate an infection which may affect fertility. Although CASA system can be used to automatically measure sperm concentration, errors are encountered when there are high and low concentrations, significant agglutinates or large amount of significant debris. The manual count can be obtained with a coefficient of variation of less than 10%.