BACKGROUND:After in vitro fertilizationand embryo transfer for tubal infertility, gamete intrafallopian transfer has been introduced for patients with non-tubal infertility. However, the gametes need to be transferred in 2 to 5 minutes and the distance between the operating theatre and tissue culture laboratory delayed its introduction at our hospital.METHODS AND PATIENTS:To overcome this problem we designed a box in which gametes could be stored. Using gametes taken from this box and employing the standard technique, we achieved 5 pregnancies in 39 infertile women.RESULTS:From 41 treatment cycles, 39 women underwent oocyte retrieval. Five pregnancies were achieved of which 4 delivered live births at full term and 1 ended in abortion. Our first gamete intrafallopian transfer baby was born on 6 January 1988.CONCLUSION:The gamete intrafallopian transfer technique can be successfullyadapted for India.

Background. After in vitro fertilizationand embryo transfer for tubal infertility, gamete intrafallopian transfer has been introduced for patients with non-tubal infertility. However, the gametes need to be transferred in 2 to 5 minutes and the distance between the operating theatre and tissue culture laboratory delayed its introduction at our hospital. Methods and Patients. To overcome this problem we designed a box in which gametes could be stored. Using gametes taken from this box and employing the standard technique, we achieved 5 pregnancies in 39 infertile women. Results. From 41 treatment cycles, 39 women underwent oocyte retrieval. Five pregnancies were achieved of which 4 delivered live births at full term and 1 ended in abortion. Our first gamete intrafallopian transfer baby was born on 6 January 1988. Conclusion. The gamete intrafallopian transfer technique can be successfullyadapted for India.

The effects of intranasal administration of norethisterone (NET) on menstrual cycle length, folliculogenesis, serum levels of estradiol, FSH, LH and progesterone, vaginal cytology, cervical mucus and endometrial morphology were studied in 8 volunteers (age 28 to 39 years, weighing between 46 to 54 kg). The study period comprised 4 consecutive menstrual cycles. In the first cycle (pretreatment cycle), only the vehicle (alcohol, propylene glycol, water; 3:3:4) was sprayed intranasally (100-mu-l in each nostril), using a metered nebulizer, once daily from day 3 to the last day of menstrual cycle. In the next two cycles (treatment cycles), NET (300-mu-g/day) was administered once daily, starting from day one of menstrual cycle, between 9 and 10 a.m. The fourth cycle was a post-treatment cycle in which the volunteers were monitored for recovery. Blood samples (about 5 ml each) were collected once daily from day 8 to 24 and thereafter on alternate days until the last day of cycle during all the 4 cycles. Levels of estradiol, FSH, LH and progesterone were measured in the serum samples by radioimmunoassay methods. Cervical mucus samples and vaginal smears were collected once daily starting from day 7 or 8 of each cycle until the mucus was very scanty. Serial pelvic ultrasonography was performed starting from day 7 or 8 until the growing follicle disappeared or throughout the cycle in case a growing follicular cyst was observed. Endometrial aspirates were collected once around day 22 in each cycle and processed for routine histological examination.Intranasal administration of NET had no significant effect on mean menstrual cycle length. However, other parameters were affected in a significant number of treatment cycles. The mean cervical mucus score of 13.5 in pretreatment cycle was reduced significantly in 5 and 6 volunteers in the first and second treatment cycles, respectively, and in the others it was not affected. In 10 treatment cycles the vaginal cytology was indicative of anovulation and in one each it was inconclusive and ovulatory. Since in 2 volunteers the vaginal smears during the pretreatment cycles were not indicative of ovulatory cycles, it was considered that their vagina were probably refractory to hormonal changes and were thus not considered for purposes of evaluating the effects of NET on vaginal cytology. On the basis of endocrine profile, 8 cycles showed ovulatory pattern in which progesterone levels were indistinquishable from pretreatment cycle, 5 cycles were anovulatory and the remaining 3 were deficient in progesterone and had longer follicular phase and shorter luteal phases. Ultrasonographic evidence of normal folliculogenesis and ovulation was observed in 7 cycles. Formation of follicular cysts was seen in 8 treatment cycles of which 7 were anovulatory and in one ovulation was observed in the contralateral ovary. The size of the cysts varied between 29 to 44 mm. The cysts disappeared either just before the menses in the same cycle or by the middle of post-treatment cycle. A positive correlation (r = 0.679) in the size of follicular cyst with estradiol levels was observed. In two cycles normal folliculogenesis was observed; however, the follicle did not rupture at the anticipated time. Since these two cycles had normal estradiol and progesterone levels, these were classified to have luteinized unruptered follicles. Out of 11 samples of endometrial aspirates, which could be collected during two treatment cycles, hormonal effects were seen in 10 and in one the endometrium was secretory. In 5 cycles where the aspirates could not be collected, it is possible that the endometrium was either very scanty or had undergone atrophy. These observations, therefore, provide convincing evidence that NET administered intranasally causes changes both in the reproductive endocrine parameters as well as on the end organs and that these changes are suggestive of antifertility effects.

Four different ovarian stimulation protocols were evaluated in an in vitro fertilisation and embryo transfer programme in 208 women (228 treatment cycles). In the rigid protocol (RP), 100 mg of clomiphene citrate (CC) was given from day 3 to day 7 of the menstrual cycle and 300 IU of human menopausal gonadotropin (hMG) was given from day 5 of the menstrual cycle. In the individualised protocol (IP) the same drugs and doses were used as in RP, but the day of initiation of CC depended on the length of the individual's menstrual cycle and hMG was administered from the last day of CC. In the programmed protocol (PP), ovarian function was suppressed with oral contraceptive pills (ethinyl estradiol 30-mu-g and norethisterone 1 mg) started on day 5 of the menstrual cycle for 45 to 70 days. Considering the last day of pill intake as day 0, CC was given for 5 days from day 5 and hMG (300 IU) from day 7. In the alternate day protocol (ADP), 100 mg of CC was administered from day 2 to day 6 and hMG (300 IU) was given on alternate days from day 2 to day 8 or day 10 of the cycle. In all the women, hCG (5000 IU) was administered when the diameter of at least 2 follicles was greater-than-or-equal-to 16 mm and estradiol levels were 300 pg/ml/dominant follicle. Patients not showing such a response were not treated further. The cardinal events of IVF-ET such as number of good responders, incidence of oocytes harvested, fertilised and embryos transferred per cycle were compared and it was concluded that the pregnancy rates were highest in women treated by the PP.

The advantages of using scanning electron microscope (SEM) vis-a-vis light microscope (LM) to assess sperm morphology was studied. The semen samples obtained from 15 fertile (group I) and 25 infertile (group II) men were processed by routine procedures for LM and SEM. The usually described sperm abnormalities were identifiable with greater resolution. Sperm abnormalities were significantly more in group II, as compared with group I (P less than 0.001) by both methods. Abnormalities of different regions, like thinning of midpiece, were scored higher in SEM than in LM. The abnormality of aggregation or retraction of mitochondrial sheath was clearly visible under SEM and missed by LM. Apart from detection of certain subtle abnormalities, SEM has advantages of rapid screening, accurate quantitation directly from the screen, zooming to higher magnification and automation.

Trichomonas vaginalis is a sexually transmitted anaerobic parasite which causes vaginitis and cervicitis. The present study was carried out to determine the incidence ofTrichomonas infection in semen samples of asymptomatic men and also to understand the changes of semen characteristics in the infected individuals before treatment and after treatment with metronidazole (Flagyl, 400 mg×3× a day for 10 days). The semen obtained from 1131 men was examined for different characteristics andTrichomonas were detected in wet smear preparations in 50 cases (4.42%). The characteristics of semen in them was compared with 52 normal semen samples. Statistical analysis showed that the seminal fluid viscosity and percentage particulate debris was increased significantly in the infected group (P<0.001). There was no significant change in the pH of semen. Spermatozoan motility and morphologically normal forms were decreased significantly (P<0.001), spermatozoan viability was altered, and there was a significant change in membrane integrity (P<0.001) as indicated by the hypoosmotic swelling test. The significant improvement in semen characteristics were seen in 25 cases after a single course of treatment. Therefore, it is possible that in some cases, the infertility seen in asymptomatic individuals may be due to infection byTrichomonas.

Two distinct types of cortical granules were discerned in the human oocyte. The first type, G1 granules, had a mean diameter of 350 nm, contained a uniformly compacted, electron-dense substance; these granules were probably synthesized even before the oocyte was aspirated from mature follicles and their contents were never found to be secreted in any of the preovulatory (immature), unfertilized and fertilized eggs examined. The G2 granules measured 450 nm (mean diameter), had a granular content and were found to be synthesized and secreted at all the stages of egg development studied. Endocytosis was also evident in the unfertilized and fertilized eggs. The secretion of the G1 cortical granules even before fertilization suggests that they may have an additional role, rather than merely contributing to the blockade of polyspermy.

Morphological and histological features of rete testis, vas efferens, epididymis and vas deferens were studied in the langur monkey. Tubular extensions of rete were located towards lateral side of the testis. Its epithelium comprises mostly of cuboidal cells with hyaline cytoplasm. Three to nine bundles of vas efferens, emerging below the cranial pole of the testis, were observed. Vas efferens epithelium comprises of ciliated and nonciliated cells. Epididymis could be divided into six zones on the basis of cytological features. Principal cells, basal cells, apical cells and intraepithelial lymphocytes were observed in the epididymal epithelium, but their number, shape, size and location of nuclei varied in different zones. Vas deferens epithelium comprises of principal cells, basal cells, apical cells and few intraepithelial lymphocytes. Epithelium is surrounded by lamina propria, longitudinal, circular and longitudinal muscle layers.

In 5 caged langurs menstrual cycle length (for 125 cycles) varied from 18 to 45 days with a mean +/- s.d. of 26.3 +/- 3.6 days. Menstruation lasted for 2 days. A distinct cycle of vaginal smear cells was observed with a maximum cornification on Day 10 coinciding with the serum oestradiol peak. A monophasic pattern of vaginal temperature with a significant nadir on Day 11, 1 day after the oestradiol peak, was observed. Serum sialic acid concentrations directly reflected the oestrogenic activity and showed a significant peak on Day 10, coinciding with the oestradiol peak and was followed by a progesterone rise. Langurs menstruated throughout the year without showing any sign of summer amenorrhoea. We suggest that maximum cornification, vaginal temperature nadir and serum sialic acid peak could be used as markers for prediction and detection of the time of ovulation in langur monkeys.

Daily sperm production in rats and monkeys were estimated by extended histometric method. Individual seminiferous tubules were separated, fixed, processed and embedded. Semi-thin sections were stained with toluidine blue and the different cell types quantitated with Chalkley’s eye piece. The daily sperm production estimated was compared with that obtained by the homogenization method. The results indicate that daily sperm production obtained by both the methods were comparable. The present histometric method has an added advantage of requiring very small amount of biopsy material to give precise daily sperm production estimations.

Pharmacokinetic parameters of norethisterone (NET) were studied in eight adult male bonnet monkeys following the administration of a single dose of 300 ug. The animals were crossed over between the following three routes of administration: oral ingestion, nasal and sublingual spraying. The results indicate that NET was readily absorbed by all three routes but the Cmax and AUC of NET were significantly greater by the sublingual route. No significant difference in the t 1/2 alpha or t 1/2 beta was observed between the three routes. These findings suggest that the sublingual route offers the possibility of reducing the effective dose of NET, which is widely used for contraceptive purposes.

The endocrine and gametogenic status of the testes were studied in 9 healthy adult rhesus monkeys of proven fertility throughout a one-year period. Testosterone levels were estimated by radioimmunoassay in blood samples collected at 4 h intervals over a 24 h period once a month. Semen samples and testicular biopsies were also examined once a month. A well-defined circandian rhythm was evident in serum levels of testosterone. The rhythmicity was less pronounced in February and September. The 24 h mean levels of serum testosterone were high between the months of August to March and low in the months of May to July. All animals did not uniformly respond to electro-ejaculation in April and May. Semen volume and total number of spermatozoa were maximal between September and March and least from April to August. Testicular biopsies indicated that all stages of spermatogenesis were evident between September and March and the spermatogenic activity was less evident between April and August. The contents of Sertoli cells showed a seasonal cyclicity; they were laden with lipid droplets during April to August when spermatogenesis was quiescent and vacuolated during September to March when spermatogenesis was active. These studies indicate that the testing of contraceptive drugs needs to be restricted to months of September to March in male rhesus monkeys otherwise, it is possible that the naturally occurring reproductive quiscence may be attributed to the effect of the drug being tested. The data accrued from the present studies also provide quantitative information on circulating levels of testosterone which could be used as a reference background while evaluating the contraceptive drug-effects in male rhesus monkeys.