Investigation of sex chromosome abnormalities in teratozoospermia of infertile men using fluorescence in situ hybridization.

Fluorescence in situ hybridization is a fast and efficient method of investigating chromosomal abnormalities in human spermatozoa. In this study, we have established the frequency of sex chromosome disomy (XX, XY and YY) in teratozoospermia (98-100% abnormal morphology) of infertile men compared with normospermic men using double-probe FISH procedures. A total of 40,000 sperms were scored in each group of semen for chromosome X and Y with overall hybridization efficiency of 97.00 per cent and 98.02 per cent in infertile men and normal men, respectively. It was found that the frequency of disomy XX, XY and YY was significantly higher in infertile men compared with normal men (P < 0.05). It is concluded that the infertile men showed increased frequencies of sex chromosomal abnormalities.

Contraceptive efficacy and adverse effects of testosterone enanthate in Thai men.

In conclusion, the present study demonstrated that induced severe oligozoospermia or azoospermia by weekly testosterone enanthate injection was sufficient for male contraception. The low rates of discontinuation due to side-effects of the hormone and incidental medical conditions in this study confirms the safety and acceptability of such androgen administration found in studies with up to 18 months exposure. The long-term hazards remain uncertain and require investigation for risk. The possible long-term benefits from androgen use for bone, muscle and blood metabolism will also need to be assessed before the net risk-benefit effects of an androgen-containing regimen can be fully evaluated. In summary, the contraceptive efficacy for male contraception in this study demonstrated that weekly injections of testosterone enanthate can provide safe and effective contraceptive protection. The practicability of this approach may be improved by the use of longer-acting testosterone preparations which are under development.

Determination of the ratio of X and Y bearing spermatozoa after albumin gradient method using double-labelled fluorescence in situ hybridization (FISH).

Determination of X and Y human spermatozoa by double labelled fluorescence in situ hybridization (FISH) is a new and more reliable method than Quinacrine hydrochloride staining for Y sperm identification. A preliminary report on determination of the ratio of X and Y sperm of normal semen of fifteen donors by double labelled FISH method before and after Y sperm separation by Ericsson albumin column method. This is currently popularized in Thailand. Results of standardized FISH technique has an average success rate of approximately 95 percent. Results of the experiment showed that the ratio of X:Y sperm before and after albumin column filtration is 50.03:49.80 and 48.77:51.00, respectively. Few spermatozoa have double sex chromosomes, XX, XY or YY. In conclusion, Y sperm selection by Ericsson's method is ineffective for male sex preselection.

Suppression of spermatogenesis by testosterone enanthate in Thai men.

To investigate the effect of testosterone enanthate on suppression of spermatogenesis in Thai men, 17 normal Thai men were given 200 mg testosterone enanthate weekly by intramuscular injection. During treatment, semen was collected regularly to monitor spermatogenesis. Median times for the first semen sample reaching sperm concentration threshold of 5, 3, 1 and 0 million/ml were 58, 70, 84, and 85, respectively. Subsequently, all men became azoospermic. Among 17 men entering the efficacy phase, 14 (82.3%) achieved consistent azoospermic from the beginning of efficacy phase, the remaining 3 (17.7%) were initially severe oligozoospermic but later became azoospermic. Only one case achieved consistent oligozoospermia but did not achieve azoospermia within 6 months. After stopping injection, sperm first reappeared in the ejaculate of formerly azoospermia men at 73 days. Recovery of sperm output to normal sperm concentration (> 20 million/ml) was achieved by all men at a median time of 3.9 months and recovery to their own baseline in one year after the last injection was established in 13/17 (76.5%) at a median time of 4.9 months, respectively. In conclusion, testosterone enanthate alone is effective in suppression of spermatogenesis for male hormonal contraception due to the high rate of azoospermia induced, which is known to ensure reliable and effective, reversible contraception.

Prediction of post-thaw sperm motility and sperm cryosurvival rate using the pre-freeze sperm parameters.

To determine the relationships between pre-freeze semen variables and cryosurvival rate and post-thaw motility and examine whether they have any predictive value for the cryosurvival rate and post-thaw sperm motility, conventional semen analysis, supravital staining for sperm viability and hypo-osmotic swelling test were performed on 50 semen samples before cryopreservation. Thawed semen samples were examined for post-thaw sperm motility and cryosurvival rate. Significant correlations were observed between post-thaw sperm motility and several pre-freeze semen variables, such as, hypo-osmotic swelling test, pre-freeze sperm motility and sperm viability. In a stepwise regression analysis, an accurate prediction of post-thaw sperm motility (R = 0.826) was obtained using a multiple regression equation incorporating 3 variables including hypo-osmotic swelling test, pre-freeze sperm motility and sperm concentration. In conclusion, a set of criteria have been identified that accurately predicts post-thaw sperm motility and which place particular emphasis on hypo-osmotic swelling test. Conventional semen analysis and hypo-osmotic swelling test are simple and effective assays for the prediction of post-thaw motility.

A comparison of Makler counting chamber and improved Neubauer hemocytometer in sperm concentration measurement.

Evaluation of male fertility is based predominantly on results from semen analysis and determination of the sperm concentration is one of the main parameters of the analysis. To assess the accuracy of sperm concentration measurements by Makler counting chamber, manual sperm counting of 55 semen samples were made using a Makler counting chamber, and compared with concentration values measured using an improved Neubauer hemocytometer according to the World Health Organization guideline (standard procedure). Results were classified according to the standard procedure before statistical analyses. Both values correlated well. Sperm concentration obtained with Makler counting chamber was not statistically different from those determined by improved Neubauer hemocytometer in semen samples with concentrations over 40 x 10(6)/ml. But using Makler counting chambers caused a shift concentrations, which were overestimated significantly (p less than 0.0001) in semen samples with concentrations less than 40 x 10(6)/ml. Overall, Makler chamber counts were 11.2 per cent higher. Although less complicated than the improved Neubauer hemocytometer method, measurement of sperm concentration by Makler counting chamber is an inaccurate method, especially in semen samples with concentrations less than 40 x 10(6).

Effects of pentoxifylline on sperm motility characteristics and motility longevity of postthaw cryopreserved semen using computer-assisted semen analysis (CASA).

Computer-assisted sperm movement analysis was used to study the effect of pentoxifylline on human sperm motility characteristics and motility longevity of postthaw cryopreserved semen. This study focused on the following issues: the changes in individual movement characteristics in response to pentoxifylline, the persistence of the response during drug treatment. Computerized analysis was started at 30 min, 3 hrs and 24 hrs, after addition of pentoxifylline. Data obtained showed that pentoxifylline significantly increased percentage of sperm motility, average path velocity (VAP), curvilinear velocity (VCL), straight line velocity (VSL), amplitude of lateral head displacement (ALH) at two of the three time periods (p < 0.05). But, it did not significantly increase linearity (LIN), straightness (STR), and beat cross frequency (BCF) at any time. After 24 hrs, all motility variables were significantly decreased in both groups. However, the 24-hr motility longevity in the treatment group was greater than that of the control group. The present analysis shows interest in the use of pentoxifylline as a sperm movement enhancer for postthaw cryopreserved semen. It shows a beneficial effect in the majority of sperm movement parameters and motility longevity which may increase pregnancy rates after insemination. However, whether this change leads to an increase in fertilizing ability requires further study.