Preimplantation genetic diagnosis of chromosomal abnormalities by multicolour fluorescence in situ hybridisation.

Preimplantation genetic diagnosis (PGD) is an early diagnosis of genetic disorders, prior to the onset of pregnancy. PGD incorporates the latest techniques in assisted reproduction and molecular genetics. Embryos or oocytes are biopsied during culture in vitro and genetic analysis is carried out on the blastomeres or polar bodies. Embryos shown to be free of the genetic disease under investigation are transferred to the uterus. Multicolour fluorescence in situ hybridisation (FISH) is used to diagnose numerical and certain structural abnormalities of chromosomes in the embryo. The common probes used are for chromosomes 13, 18, 21, X and Y. FISH can also be used for PGD of translocations, when one of the parents is a carrier. PGD was carried out recently in 4 cases using multicolour FISH. In one of the embryos, trisomy 18 was detected. Tetraploidy was seen in another embryo. Only chromosomally normal embryos were transferred back to the uterus. Care has to be taken while interpreting FISH signals as the signal may be split, diffused, superimposed or in a different focus.

Genital tuberculosis--a major pelvic factor causing infertility in Indian women.

OBJECTIVE: To study the effect of tuberculosis, a common infectious disease in the Indian subcontinent, on the female pelvic factor and its subsequent effect on female fertility. DESIGN: Retrospective case studies. SETTING: Department of Infertility Management and Assisted Reproduction, Jaslok Hospital and Research Centre, Bombay, India. PATIENT(S): Three hundred women, between the ages of 25 and 35 years, with tubal factor as a cause of their infertile state. RESULT(S): One hundred seventeen women with a tubal factor were found to have tuberculosis as the cause of tubal blockage. On laparoscopy, 49.5% were found to have simple tubal blockage, 15.3% showed tubo-ovarian masses, and 23.9% had a frozen pelvis. Seventy-five percent complained of menstrual irregularities, thus indicating endometrial involvement; 25.6% of these women underwent an IVF procedure. The pregnancy rate after IVF-ET was 16.6% per transfer. CONCLUSION(S): This study highlights the fact that tuberculosis, a chronic infectious disease, is one of the major etiologic factors of female tubal infertility, especially on the Indian subcontinent.

Computer-assisted semen analysis parameters in men with varicocele: is surgery helpful?

OBJECTIVE: To assess sperm characteristics and fertility before and after varicocelectomy using computer-assisted semen analysis (CASA). DESIGN: Preoperative and postoperative sperm parameters of infertile men with varicocele were analyzed statistically and the outcome of pregnancy was determined. SETTING: Department of Infertility Management and Assisted Reproduction, Jaslok Hospital and Research Centre, Bombay. PATIENTS: Semen samples of 49 men with varicocele-related infertility were analyzed using CASA. INTERVENTIONS: Sperm parameters of 26 men who underwent varicocelectomy were evaluated 3, 6, and 9 months postoperatively. Conception, occurring either naturally or therapeutically, was recorded. MAIN OUTCOME MEASURES: Evaluation of improvement of sperm parameters after surgery. RESULTS: Sperm count, motility parameters, curvilinear velocity, straightline velocity, lateral head displacement, and normal morphology were significantly lower in men with varicocele. Postoperatively, there was significant improvement in count, motility, and normal morphology, with a decrease in proportion of acrosome-deficient heads and tapering forms. After varicocelectomy, 46.2% of the men had normal semen parameters, with the overall pregnancy rate being 50%. CONCLUSIONS: Computer-assisted semen analysis provides the potential for accurate quantitative evaluation of semen in men with varicocele. Varicocelectomy results in improvement in semen quality with pregnancy rates of 50%.

Assisted hatching in an in vitro fertilization programme.

Assisted hatching was performed on 37 women (IVF, n = 12; micromanipulation, n = 25), who were at risk of poor implantation after in vitro fertilization and micromanipulation, and who had previous cycles of failed implantation. A total of 292 oocytes was retrieved (7.9 per cycle) of which 90 were subjected to IVF and 202 were micromanipulated. Assisted hatching was performed on day 3 of culture, on six- to eight-cell embryos, by the technique of mechanical partial zona dissection. All the patients undergoing the procedure of assisted hatching had embryos with thick zonae, greater than 15 microns. Assisted hatching was performed on 39 embryos (55.7%) in the IVF group and 104 embryos (77.6%) in the micromanipulation group. The damage rate after hatching was 1.4%. The pregnancy rate per cycle was 50% in the IVF group and 44% in the micromanipulation group, compared with 10% (IVF) and 19.2% (micromanipulation) in the control groups. Assisted hatching, by partial zona dissection, results in significant improvement in pregnancy rates in women more than 38 years of age, who have a thick zona and with previous poor implantation results. This paper reports the first pregnancy in India by the technique of assisted hatching (partial zona dissection).

Multinuclear spermatozoa associated with polyploidy.

We are reporting on a possible association between multinucleated spermatozoa and polyploidy after in vitro fertilization. The semen from the male partner of a couple with 40% polyploidy on three in vitro fertilization attempts was examined by light and transmission electron microscopy. In that sample 10% to 12% of spermatozoa demonstrated conjoined or double heads. The significance of these findings is discussed.