explore the possible presence of varicocele. Semen samples were obtained from both fertile and infertile men. Weight, height and waist circumference of all participants were measured. The body mass index (BMI) was calculated as weight ⁄height2 (kg/ m2) and subjects were classified into: normal weight (BMI 18–25 kg/ m2), overweight (BMI > 25–30 kg/ m2) or obese (BMI > 30 kg/ m2). Waist circumference was measured at the mid-point between the lower borders of the rib cage and the iliac crest. Semen Analysis: Standard semen analysis was performed according to the guidelines of the fifth edition of the WHO laboratory manual for examination of human semen.19 Participants were asked to deliver a semen sample (by masturbation) into a sterile plastic container after 2-7 days of sexual abstinence. The samples were left to liquefy in a 37°C incubator, and were analyzed within one hour of delivery. Standard procedures included …show more content…
21 Fresh semen samples were diluted with phosphate-buffered saline (PBS) to 2x106 sperm/mL. Fifty µL were incubated with 100 µL lysing reagent for 15 seconds and then 2 mL of PI was added and mixed with tube. Tube acquisition was done by flowcytometry immediately after staining. The intensity of its emission corresponds to the DNA content. Flowcytometric analysis displays a constant and characteristic bimodal nonartifactual DNA pattern indicating the presence of two different populations. The main population is represented by a peak followed by a shoulder ("marginal population"), which represents a sperm group altered in the nuclear condensation, yielding unstable chromatin which appears more stainable with PI. The percentage of sperm cells with DNA damage was automatically calculated after acquisition