The outcome of sperm retrieval and intracytoplasmic sperm injection for obstructive azoospermia.

OBJECTIVES: To study the outcome of sperm retrieval and intracytoplasmic sperm injection (ICSI) from obstructive azoospermic men. METHOD: Overall, 50 sperm retrieval procedures were performed in 45 obstructive azoospermic men, followed by 57 ICSI procedures with fresh epididymal spermatozoa (n=40), fresh testicular spermatozoa (n=4) or frozen-thawed epididymal spermatozoa (n=13). RESULTS: Sperm retrieval was accomplished via percutaneous epididymal sperm aspiration (PESA) in 42 cases, testicular sperm aspiration (TESA) in 1 case and testicular sperm extraction (TESE) in 2 cases. TESA and TESE were only applied when PESA failed to produce enough spermatozoa for simultaneous ICSI. PESA was successful in 92 per cent of cases. Fertilization rate after ICSI was 79.6 per cent of the metaphase II oocytes. Seventy one embryo transfers were performed using both fresh and frozen thawed embryos resulting in clinical pregnancy in 39.4 per cent. Ongoing pregnancy was achieved in 35.2 per cent. CONCLUSION: ICSI has been shown to give a high fertilization and pregnancy rate with epididymal and testicular spermatozoa retrieved from obstructive azoospermic men. PESA is a noninvasive and simple technique for retrieving spermatozoa from obstructive azoospermic men. Therefore, it is suitable as the primary sperm recovery technique in patients with obstructive azoospermia.

Comparison of the outcome of intracytoplasmic sperm injection using fresh and frozen-thawed epididymal spermatozoa obtained by percutaneous epididymal sperm aspiration.

The study was conducted to compare the outcome of intracytoplasmic sperm injection (ICSI) between fresh and frozen-thawed epididymal spermatozoa retrieved by percutaneous epididymal sperm aspiration (PESA) from obstructive azoospermic men. Overall, 53 PESA procedures were performed in 42 obstructive azoospermic men, followed by ICSI procedures with either fresh (n = 40) or frozen-thawed (n = 13) epididymal spermatozoa. Comparing all ICSI cycles with fresh and frozen-thawed epididymal spermatozoa, the fertilization rates (77.4 vs 86.8%) and the cleavage rate (91.3 vs 95.1%) were not statistically different. A total of 64 embryo transfers were performed: 48 embryo transfers after the use of fresh epididymal spermatozoa and 16 embryo transfers after the use of frozen-thawed spermatozoa. The overall pregnancy rate per embryo transfer was also similar between both groups (40.8 vs 40%). The implantation rate per embryo (18.2 vs 12.7%), clinical pregnancy per embryo transfer (36.7 vs 33.3%) and delivery/ongoing pregnancy rate (36.7 vs 33.3%) were not statistically different. In conclusion, there were no significant differences of the outcome of intracytoplasmic sperm injection using fresh and frozen-thawed epididymal spermatozoa obtained by PESA.

Infected ovarian cyst in a homozygous beta-thalassemic patient.

An infected ovarian cyst in a thalassemic patient is rarely reported. We describe the case of a 22-year old woman with splenectomized homozygous beta-thalassemia who developed high fever and was diagnosed as having an infected ovarian cyst. The mechanisms which beta-thalassemia might predispose to infection and considered to be immunocompromized are discussed. She was given an intravenous antibiotic regimen and the infected ovarian cyst was removed. The difficulties in the diagnosis of an infected ovarian cyst is because of its rarity and the paucity of information on it in the literature. Therefore, the triad of ovarian cyst, immunocompromized host, and signs of infection with failure to identify any other source of infection should raise the suspicion of an infected ovarian cyst.

Comparison of the fertilization rate after intracytoplasmic sperm injection (ICSI) using ejaculated sperms, epididymal sperms and testicular sperms.

Intracytoplasmic sperm injection (ICSI) has been successfully used to achieve fertilization and pregnancies for patients with extreme oligoastheno-zoospermia using ejaculated sperms or patients with azoospermia using epididymal or testicular sperms. The aim of this study was to compare the fertilization rate after ICSI using ejaculated, epididymal and testicular sperms. Between January and September 1997, 10 azoospermic men underwent percutaneous epididymal sperm aspiration (PESA) or testicular sperm extraction (TESE) to recover sperm for ICSI. A total of 5 PESA cases and 5 TESE cases were performed at the Center for Assisted Reproduction & Embryology. Thirty-one patients performed ICSI using ejaculated sperms during the same period of time were used as a control group. ICSI using ejaculated sperms, epididymal sperms from PESA and testicular sperms from TESE was a highly successful technique, achieving fertilization rates of 78.5 per cent, 83.3 per cent and 80.8 per cent, respectively. Good fertilization rates were achieved without significant differences among the various sperm sources.

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.

Effects of occupational lead exposure on spermatogenesis.

In summary, there were significant differences in blood and semen lead levels, LH levels and prolactin levels in workmen who were exposed to lead. However, all hormonal profiles still remained within the normal adult range for Thai men. Higher levels of blood lead were found to have no significant effect on the sperm variables and hormonal profiles except testosterone levels at second follow-up and prolactin levels at sixth follow-up. However, these differences were not consistent during the study period. It should be noted that the present data are preliminary, pertain only to short-term exposure and require confirmation. The actual prospective long-term risk of lead exposure, remains to be determined. However, our study highlights the feasibility and importance of a prospective cohort study in assessment of the risk of lead exposure on male reproductive system. Long-term longitudinal controlled studies of semen quality and hormonal profiles in workmen examined before and during lead exposure and prospective studies of time to pregnancy in couples attempting to achieve pregnancy are probably necessary in order to obtain further knowledge. This study also indicates that the reproductive function surveillance of these workmen must be investigated as well as other physical health. In conclusion, an increase of blood lead levels due to occupational exposure cause no immediate and significant changes in spermatogenesis and hormonal profiles.

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.

Evaluation of the Hamilton-Thorn Motility Analyzer for routine semen analysis.

In routine semen analysis of 102 patients the values of sperm concentration and the percentage of sperm motility were measured with the Hamilton-Thorn Motility Analyzer (HTMA) and compared to the data obtained by conventional semen analysis according to the guidelines of the World Health Organization (WHO). Results were classified according to the conventional semen analysis before statistical analyses. Overall, sperm concentration by both methods showed a correlation of 0.95 (p < 0.0001) but the HTMA gave higher values for sperm concentration (mean difference 10.8 +/- 18.9 x 10(6)/ml, mean +/- S.D.). Only a sperm concentration of between 40 and 80 x 10(6)/ml can be measured accurately with the HTMA. The percentage of sperm motility by both methods showed a correlation of 0.759 (p < 0.0001) but were estimated slightly lower by the HTMA than by conventional analysis (mean difference 1.7 +/- 15.8%). In conclusion, the HTMA system can not replace conventional semen analysis in routine semen analysis.

Evaluation of sperm nuclear DNA normality by acridine orange staining technique.

To determine the relationship between sperm deoxyribonucleic acid (DNA) normality and fertilizing potential, 93 semen samples from 48 fertile donors and 45 male partners of infertile couples whose major abnormalities in the female partner had been ruled out were studied. Semen samples were assessed for conventional parameters (volume, percentage of normal morphology, percentage of progressive motility, sperm concentration, and round cell concentration) according to World Health Organization (WHO) guideline and acridine orange staining. The mean sperm concentration, percentages of progressive motility, and percentage of green fluorescing sperm in semen were significantly higher in samples from fertile donors (p < 0.005). When the parameters of semen quality were considered normal according to the standard WHO criteria, the mean percentages of green-fluorescing sperm were significantly higher in fertile donors than infertile patients (65.6% vs 53.3%; p < 0.05). Therefore, the acridine orange staining technique for evaluation of the sperm DNA normality appears to give more information in infertile patients with normal semen analysis. It may be a useful addition to the conventional semen analysis.

The effect of discontinuous Percoll gradient centrifugation on sperm morphology and nuclear DNA normality.

To determine the improvement of sperm morphology and sperm nuclear DNA normality after discontinuous Percoll gradients preparation, sperm morphology and sperm nuclear DNA normality from 157 semen samples from 45 donors and 63 male partners of infertile couples were assessed before and after discontinuous Percoll gradients preparation. In semen samples, about 67.5 per cent of the nuclei exhibited green acridine orange fluorescence. The percentage of normal sperm morphology, percentage of progressive motility, and percentage of green-fluorescing sperm were significantly better after this sperm preparation. Therefore, this sperm preparation technique is a convenient and efficient method to select progressive motile, normal morphological and normal nuclear DNA sperm for assisted conceptive technology.

Efficiency of discontinuous Percoll gradients preparation for selection of motile sperm from poor quality semens.

Efficiency of discontinuous Percoll gradients preparation for selection of motile sperm from poor quality semens was studied in 106 semen samples, of which 52 were normozoospermia, 9 were oligozoospermia, 29 were asthenozoospermia, and 16 were oligoasthenozoospermia. Sperms from each semen sample were recovered by the discontinuous two-layer (50%/100%) Percoll centrifugation technique. Overall motile sperm recovery rate was about 27 per cent. The results obtained in this study indicated that sperm selection by this method from oligozoospermic and oligoasthenozoospermic semen samples was not as good as normozoospermic semen samples because per cent progressive motility showed no statistically significant improvement after sperm selection in oligozoospermic and oligoasthenozoospermic semen samples. Moreover, total sperm recovery rate and motile sperm recovery rate of oligozoospermic semen samples were less than normozoospermic samples significantly (p < 0.01 and p < 0.005, respectively).

The effect of discontinuous Percoll gradients preparation on sperm movement characteristics.

To determine the improvement of sperm movement characteristics after discontinuous Percoll gradients preparation, sperm movement characteristics from 60 normal semen samples were assessed before and after discontinuous Percoll gradients preparation using Hamilton-Thorn Motility Analyzer (HTMA). There was a significant correlation between manual and HTMA results for the percentage of motile sperm (r = 0.8670, p < 0.001). The percentage rapid, percentage motile, average path velocity (VAP), progressive velocity (VSL), curvilinear velocity (VCL), beat cross frequency (BCF), amplitude of head displacement (ALH), percentage of progressive motility, and hyperactivation were significantly increased after preparation. Therefore, this sperm preparation technique is a convenient and efficient method to prepare progressive motile sperm for assisted conceptive technology.

Sonographic prenatal diagnosis of heterozygous achondroplasia: a case report.

A case of heterozygous achondroplasia diagnosed prenatally by sonographic examination at third trimester, demonstrating a large head, frontal bossing, low nasal bridge, shortening of the long bones (rhizomelic form), was presented and confirmed by neonatal physical examination and radiograph. Sonography has become the most sensitive and accurate method for detection of this abnormality. It displays the fetal biometry without distortion and permits accurate prenatal diagnosis. However, the prenatal diagnosis of this abnormality is very difficult until the third trimester of pregnancy.

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.

Effects of digital cervical examinations on duration of latency period, maternal and neonatal outcome in preterm premature rupture of membranes.

The medical records of 115 singleton pregnancies by preterm PROM from Chulalongkorn Hospital that met the criteria for expectant treatment from January 1987 to December 1991 were reviewed. All cases were divided into 2 groups. Fifty cases had a digital cervical examination to assess cervical status and sixty-five cases had a sterile speculum examination only. Latency period, maternal and neonatal outcome of both groups were compared. There was no statistically significant difference between the latency period of both groups (34.19 +/- 78.35 versus 54.6 +/- 127.01 hours). This was consistent for all gestational age groups. There was statistically significant increased incidence of chorioamnionitis in patients receiving digital cervical examinations. A nonsignificant trend toward an increased incidence of endometritis was also noted in patients receiving digital cervical examinations. There was no statistically significant differences in the average time of delivery, mode of delivery and postpartum hospitalization between both groups. No maternal mortality and stillbirth was noted in this study. There were no statistically significant differences in Apgar score at 1 minute and 5 minute, NICU admittance, neonatal hospitalization and percentage of neonatal deaths, livebirths, and neonatal morbidity between both groups. The results of this study suggest that digital cervical examinations in patients with preterm PROM should be avoided until the clinician is convinced that the patient is clearly in labor because digital cervical examinations increase the incidence of chorioamnionitis and endometritis but have no effect on latency period.

Prenatal sonographic diagnosis of meconium peritonitis: a case report.

A case of meconium peritonitis that was diagnosed ultrasonographically in the third trimester is presented. Fetal ascites, intra-abdominal calcification, left communicating hydrocele, and polyhydramnios were detected on antenatal ultrasonography. Specks of calcification were also demonstrated on abdominal radiography postnatally. Laparotomy confirmed the diagnosis of perforated terminal ileum with meconium peritonitis. The obstetric and neonatal implications of meconium peritonitis are discussed with literature review.

Transvaginal sonographic features in ectopic pregnancy.

A retrospective review of the transvaginal sonograms of 37 women with confirmed ectopic pregnancy was performed to characterize the findings from transvaginal sonography and correlate operative findings with sonographic features. Thirty-six of the 37 ectopic pregnancies were tubal pregnancies. Intrauterine findings showed an absence of gestational sac in all cases and a pseudosac in 1 case (7.14%). An ectopic gestational sac with a live embryo was documented in 2 cases (5.41%) and with embryo or yolk sac in 6 cases (16.22%) and nonspecific adnexal mass in 30 cases (81.08%). Fluid in the cul-de-sac was found in 22 cases (59.46%). Transvaginal sonography provides better access to the pelvic organ and allows a clear view of the uterus and adnexa, with no need to fill the bladder. Excellent visualization of the endometrial cavity permits the detection of even a very early intrauterine pregnancy and clearly shows the gestational sac where it is outside the uterine cavity. Blood and other fluids in the cul-de-sac can also be seen easily. This technic has reduced patients' risk by minimizing delay in diagnosis and treatment.