Tiziana Rossi

Superoxide anion in human semen related to seminal parameters and clinical aspects

OBJECTIVE To study the possible presence of extracellular superoxide anion (O2-) in untreated semen of normal and pathological subjects and to determine the possible relationship between superoxide anion levels, number of seminal leukocytes (WBCs) and seminal parameters. PATIENTS Semen samples were analyzed from fertile normospermic subjects (n = 20) and patients (n = 132) attending the Outpatient Department of the 5th Medical Clinic. MAIN OUTCOME MEASURE We evaluated superoxide anion levels by monitoring the reduction of cytochrome c. RESULTS O2- was detected in 55.0% of fertile normospermic subjects. Both the percentage of subjects in whom O2- was detected and the mean values +/- SD of O2- were significantly higher in the male partners of infertile couples, in subjects with varicocele, and in subjects with positive sperm culture than normospermic subjects. No variations were seen in vasectomized subjects or in patients with past cryptorchidism. A high correlation was observed between O2- levels and WBCs (rs = 0.58), nonrapid, immotile sperm (rs = 0.64), percentage decrease of motility within a specified time frame (2 to 6 hours) (rs = 0.73), and sperm abnormalities (rs = 0.49). CONCLUSIONS Superoxide anion in untreated semen is produced by WBCs and atypical and nonrapid, immotile sperm. A significant increase of superoxide anion was detected in the male partners of infertile couples and in some selected andrological conditions. The toxic effect of superoxide anion on sperm cells is dependent on the time of exposure and superoxide anion concentration.

Improved human sperm recovery using superoxide dismutase and catalase supplementation in semen cryopreservation procedure.

The aim of this work was to evaluate the effects of ROS scavenger supplementation in human semen samples undergoing cryopreservation procedures.After screening out andrological pathologies, we selected 25 male partners of infertile couples with the following semen profile: volume ≥ 2.0 ml, normal viscosity, sperm count ≥20 × 106/ml, straight progressive motility (classes 1 and 2) ≥ 40% (Mazzilli, Rossi, Delfino and Nofroni (1999) Andrologia 31: 187–194), atypical forms ≤ 70%, WBCs < 1.0 times 106/ml.After liquefaction, standard semen analysis and the Hypoosmotic Viability Test (HVT) were carried out; the samples were then divided into 4 aliquots. The first was untreated as a control; 200 U/ml of superoxide dismutase (SOD) was added to the second, 200 U/ml of catalase to the third and both SOD (100 U/ml) and catalase (100 U/ml) were added to the fourth aliquot. Each aliquot was mixed (v/v) with TEST yolk buffer freezing medium (Irvine Scientific) and then frozen at −196°C. The percent recovery of progressive motile and swollen spermatozoa was evaluated after thawing.No significant variation in the recovery of progressive motility was seen in the aliquots with added SOD or catalase alone, compared to the control group. On the other hand, a significant improvement in sperm parameter recovery was seen in the aliquot with both SOD and catalase supplementation; perhaps because of their combined and simultaneous action on superoxide anion and hydrogen peroxide. These results suggest that, in some selected cases, SOD and catalase supplementation can contribute greatly to the prevention of sperm membrane lipid peroxidation by ROS and thus allow good sperm parameter recovery after freezing–thawing procedures.

Human semen hyperviscosity: prevalence, pathogenesis and therapeutic aspects

The aims of this study were (a) to determine the prevalence of subjects with semen hyperviscosity (SHV) in a large population of male partners of subfertile couples; (b) to identify any correlation between SHV and infections or inflammation of the genital tract; (c) to assess the effects of therapeutic approaches for treating SHV; and (d) to assess sperm kinetic parameters after successful treatment of SHV. A retrospective study of 1 833 male partners of subfertile couples was conducted. Next, clinical, seminal, bacteriological and ultrasound studies involving 52 subjects suffering from SHV were performed, and the SHV was classified as being mild (length of thread > 2 cm and <or= 4 cm), moderate (> 4 cm and <or= 6 cm) or severe (> 6 cm). The prevalence of SHV was observed in 26.2% (480) of the subjects, with 13.2% suffering from mild, 6.6% from moderate and 6.4% from severe SHV. Treatment was completely successful in only 27 subjects (52.0%), primarily in those who had mild basal SHV with a positive semen culture. In these subjects, progressive motility percentage, straight line velocity and linearity were significantly higher than pre-treatment levels. SHV is often found in subjects with subfertility. Pathogenesis was strictly related to infective/inflammatory factors in only 48.0% of cases; therefore, it is possible that biochemical, enzymatic or genetic factors have a role in this condition.

Superimposed Image Analysis System (SIAS) software: a new approach to sperm motility assessment.

OBJECTIVE To apply Superimposed Image Analysis System (SIAS) software (Delta Sistemi, Rome, Italy) to the objective analysis of sperm motility. DESIGN To test a new procedure for sperm motility assessment based on image superimposition. RESULTS The system can superimpose six sequential frames onto a monitor producing a final image with a motion effect similar to the multiple exposure photography method, but bypassing the film processing stage. Viewing the final image of a complete series of six superimposed frames allows the evaluation of the percentage of motile spermatozoa and their kinetic characteristics. Compared with multiple exposure photography, SIAS has the following main advantages: [1] the procedure is quick; [2] projecting the first frame alone allows the easy evaluation of sperm concentration; and [3] the system allows quick and easy analysis and measurement of each sperm trajectory frame by frame, thus avoiding misleading interpretations because of crossing sperm trajectories, collisions, and overlapping of motile spermatoza with round cells or immotile spermatozoa. CONCLUSIONS Superimposed image analysis is a new approach to sperm motility assessment. It seems to be useful particularly in selected situations, such as oligozoospermia or in cases of increased round cell concentration. In these situations, SIAS can even serve as a support or an alternative to the computer-aided sperm analysis system widely used for objective sperm analysis.

The application of the differential pH method to the biochemical evaluation of seminal plasma

OBJECTIVE To apply the differential pH method to the biochemical evaluation of seminal plasma. DESIGN AND METHODS Fructose, citric acid and free L-carnitine concentration in human seminal plasma were measured by the differential pH method. These are routinely taken as markers of seminal vesicle, prostate and epididymal function, respectively. The pH differential method was then compared with standard spectrophotometry. RESULTS The linearity, recovery and imprecision calculation for each assay were found to be reliable and the method correlated well with the reference spectrophotometric technique. In addition, the pH differential method showed certain advantages compared to spectrophotometry: a) speed of procedure (40 to 120 sec/assay); b) ability to measure the analytes even in small volumes of seminal plasma (10 microL to 50 microL) without deproteinization or dilution, thus eliminating manual operations. CONCLUSIONS The differential pH method is an innovative approach to the biochemical assessment of seminal plasma.