Cryodamage on sperm chromatin according to different freezing methods, assessed by AO test.

OBJECTIVE: To evaluate cryodamage effects on human sperm chromatin, motility and cryosurvival rate after freeze-thawing, compared between liquid nitrogen vapour and computerized program freezer, assessed by acridine orange staining method (AO test). MATERIAL AND METHOD: Fifty semen samples were used. After semen analysis, each semen sample was mixed with cryoprotective media and divided into 2 straws. The first straw was frozen with liquid nitrogen vapour and the second with computerized program freezer. After 1 month of cryostorage, semen samples were thawed. Sperm chromatin integrity, motility, morphology, vitality and sperm cryosurvival rate were determined. RESULTS: DNA damage was significantly greater (p < 0.001) following freezing with liquid nitrogen vapour than with computerized program freezer. Furthermore, the computerized program freezing method significantly provided superior post-thaw sperm motility, vitality and cryosurvival rate, compared with the liquid nitrogen vapour freezing method. CONCLUSION: Computerized program freezing is recommended as a first choice method for routine cryostorage.

Correlation of oocyte morphology with fertilization rate and embryo quality after intracytoplasmic sperm injection.

To determine whether the morphology of the oocyte assessed under light microscopy is related to the results of intracytoplasmic sperm injection (ICSI). 135 ICSI oocytes from 15 patients were analyzed. Transvaginal ultrasound-guided oocytes retrieval with oocyte morphology evaluation based on shape of the first polar body and perivitelline space followed by ICSI. After 48 hours, embryo quality was evaluated and compared to each pre-injected oocyte morphology. Normal fertilization was achieved in 81.5 per cent of the oocytes (110/135). Abnormal fertilization (3 pronuclei) was 1.5 per cent (2/135). Fertilization rate of oocytes with good morphology was higher than those with poor morphology, but there was no statistical significance (82.4% vs 79.5%; p > 0.05). Oocytes with good morphology were significantly fertilized to be embryos with good quality (p < 0.001). This study suggests that oocyte morphology correlates with embryo quality after ICSI.

Timing of second polar body extrusion and pronuclear formation after intracytoplasmic sperm injection (ICSI).

To determine timing of second polar body extrusion and pronuclear formation to find out when the intracytoplasmic sperm injected (ICSI) oocytes should be checked for successful fertilization. 148 oocytes from 15 patients from January to May 1998 were analyzed. Metaphase II oocytes were followed by ICSI. Fertilization was checked at 2, 4, 6, 8, 16 and 18 hours later for observation of second polar body extrusion and pronuclear formation. Normal fertilization was achieved in 73.6 per cent of the oocytes (109/148). After ICSI at 2, 4, 6, 8, 16 and 18 hours, extrusion of second polar body was 8.3 per cent, 49.5 per cent, 84.4 per cent, 98.2 per cent, 98.2 per cent, 98.2 per cent and pronuclear formation was 0 per cent, 0 per cent, 8.3 per cent, 30.3 per cent, 96.3 per cent, 100 per cent respectively. The earliest extrusion of second polar body and pronuclear formation were at 2 and 6 hours after ICSI respectively. This study suggests that the appropriate time to determine fertilization is at 2, 6 and 18 hours after ICSI.

ROSI from TESE the first case in Thailand: a case report.

This is a report of a success in the treatment of male infertility in a case of azoospermic male patient. Testicular sperm extraction technique (TESE) was used and round cell spermatids were retrieved. In vitro fertilization was carried out using round cell spermatid injection technique (ROSI). The zygotes were laparoscopically transferred into right fallopian tube and intrauterine pregnancy was achieved. Cesarean delivery was performed at thirty eighth week of gestation yielding a healthy male infant weighing 2820 g. The child's physical and mental developments were normal after 2 years of follow-up.

The effects of depot leuprorelin on IVF.

The trial studied the effects of depot leuprorelin on the IVF cycle and was done on nine couples. A single intramuscular injection of depot leuprorelin was given to the woman a couple days before ovulation. Seven days after ovulation, the serum progesterone level was measured and showed the same normal level as the natural ovulatory cycle. The progesterone levels varied from 12.59 to 96.0 ng/ml. On day three of the menstruation, the hormonal profiles showed a complete pituitary and ovarian suppression. FSH, LH and estrogen levels were less than 4.1 mIU/ml, 2.8 mIU/ml and 9.4 pg/ml respectively. The hMG stimulation took 11 days on average (9-15 days). A hundred and two oocytes were retrieved and among these there were 86 mature oocytes (84.3%). All oocytes were inseminated despite prematurity and resulted in 82.35 per cent fertilization. Normal fertilization occurred in 77.45 per cent (79/102). Good embryos developed in 58.23 per cent (46/79). No more than three embryos were transferred. Four women conceived, among them there was a set of twins. The implantation rate was 44.44 per cent (4/9). One abortion was found in the early first trimester. The take home baby rate was 33.33 per cent (3/9).

Comparison of pronuclear stage embryo transfer and in vitro fertilization-embryo transfer following intracytoplasmic sperm injection.

To determine whether the mode of embryo transfer (PROST vs IVF) affected the outcome in intracytoplasmic sperm injection (ICSI) cycles, 237 ICSI cycles (106 PROST and 131 IVF) were analyzed. Several parameters, including patient age, duration of infertility, amounts of hMG used, number of mature eggs retrieved and injected, fertilization rate, number of embryos transferred, and clinical pregnancy rate, were compared. Most of the variable factors were not significantly different, except the mean numbers of transferred embryos which were significantly higher in the PROST group. The clinical pregnancy rate showed no statistical difference between PROST and IVF cycles (25.5 and 16.8%; p = 0.139). This study suggests that even the pregnancy rate in PROST cycles was slightly higher than IVF cycles, but there was no statistically significant difference between the two groups.

Feto-maternal transfusion in normal pregnancy and during delivery.

Feto-maternal transfusion was studied in 2,000 pregnant Thai women. The technic used in the study was modified acid election test. The population were divided into five group, first trimester, second trimester, third, during delivery and non pregnant women. Over all fetomaternal transfusion occurred in 1,055 women (65.94%). In first trimester, second trimester, third trimester and during delivery were 53.5 per cent, 63.0 per cent, 71.1 per cent and 75.7 per cent respectively. The data were significantly different from non pregnant women (p < 0.0001). The volume of transfusion was varied from 0 to 12.65 ml and greater in the advancing gestational age. In first trimester, second trimester, third trimester and during delivery the volume of transfusion were 0.07, 0.08, 0.13, 0.19 ml respectively.