Mouse blastocyst vitrification compared with the conventional slow-freezing method.

To compare mouse blastocyst survival after cryopreservation with vitrification and the slow-freezing method, one-hundred and forty-eight in vitro mouse blastocysts were randomly frozen by the two methods: vitrification and conventional slow-freezing. After being thawed, the blastocysts were assessed for survival and hatching rate. The survival rates of blastocysts cryopreserved by vitrification and slow-freezing were 68.33 and 65.52 per cent (p = 0.89), whereas hatching rates were 51.22 and 44.74 per cent, respectively (p = 0.64). Therefore, vitrification of blastocyst-stage-embryos may be a useful, economic method for freezing the excess blastocysts in some centers where blastocysts are routinely transferred.

The regulation of assisted reproductive technology in Thailand.

The Executive Board of the Medical Council of Thailand has set up an ad hoc committee to establish the regulations of practising of assisted reproductive technology. The committee assigned the Royal Thai College of Obstetricians and Gynecologists to supervise and take charge of the administration and formulate an annual report in accordance with the Thai Medical Council Declaration. The regulation was finally approved on October 9, 1997. It was announced in the Royal Gazette on December 26, 1997 and since then the prescription of standard measures for ART practice has been effected.

Intrapartum and neonatal zidovudine treatment in reduction of perinatal HIV-1 transmission in Bangkok.

OBJECTIVE: To evaluate the efficacy of zidovudine (ZDV) administered during labor and to the infants in the first 6 weeks of life in reduction of perinatal HIV-1 transmission. DESIGN: Open label clinical trial. SITE: King Chulalongkorn Memorial Hospital, Bangkok, Thailand. MATERIAL AND METHOD: One hundred asymptomatic, antiretroviral naive HIV-1 infected pregnant women who had either late or no prenatal care were recruited from the obstetric service of King Chulalongkorn Memorial Hospital, Bangkok, Thailand. They were given ZDV 300 mg orally every 3 hours during the intrapartum period until delivery. ZDV syrup 2 mg/kg orally every 6 hours were given to the infants immediately after birth for 6 weeks. Breast feeding was not allowed. Infant's blood for HIV-1 PCR test was obtained at age 1 day, and 1, 3 and 6 months. HIV-antibody test was determined at age 18 months. Infants with at least one positive HIV-1 PCR test performed at or after 1 month of age or positive HIV-antibody test at age 18 months were classified as HIV-1 infected infants. RESULTS: There were 100 healthy infants delivered without complication. Fourteen infants were excluded due to; 13 lost to follow-up and 1 drug intolerance. Of the remaining 86 infants who were followed-up, 27 infants (31.4%) did not receive intrapartum ZDV treatment and 9 infants were HIV-1 infected. The perinatal transmission rate was 10.5 per cent, (95% CI 3.9, 17.1). CONCLUSION: The result of this study suggests that intrapartum oral ZDV treatment in asymptomatic HIV-1 infected mothers together with ZDV treatment in the neonates for 6 weeks can reduce the rate of perinatal HIV-1 transmission. This regimen may be an alternative treatment for prevention of HIV-1 infection in infants born to HIV-1 seropositive mothers who have had either late or no prenatal care.

Correlation between human follicular diameter and oocyte recovery, metaphase II oocytes and fertilization rate in intracytoplasmic sperm injection programs.

This prospective study was undertaken to establish the correlation between the follicular diameter, oocyte recovery, metaphase II (MII) oocyte, and fertilization rate in intracytoplasmic sperm injection (ICSI) programs. Thirty one patients undergoing 31 ICSI cycles from August 1998 to January 1999 at King Chulalongkorn Memorial Hospital were studied. The patients were stimulated with either short or long GnRHa portocol. During transvaginal aspiration of follicles, follicular diameters were measured. The maturation stage of oocytes and fertilization rate were evaluated. Four hundred and seventy eight follicles were measured and classified into 3 groups, group A (<10 mm), group B (10-14 mm) and group C (>14 mm). The oocyte recovery rate and MII oocytes were different between groups A, B, and C but the fertilization rate was not different. In conclusion, the follicular diameter correlated with oocyte recovery and oocyte maturation but not with the fertilization rate in ICSI programs. The quality of embryos was lower in oocytes from small follicles.

Effects of contaminated endometriotic contents on quality of oocytes.

The mechanism of infertility associated with endometriosis is poorly understood. There is evidence supporting that women with ovarian endometriosis have a lower pregnancy rate than women with peritoneal lesions only. This study aimed to evaluate the effects of endometriotic contents contamination while retrieving oocytes on oocytes' quality. Thirty-eight infertile patients with endometriotic cysts from January 1993 to June 2000 were enrolled in this study. There were no statistically significant differences among the quality of oocytes and embryos from the contaminated, non-contaminated, and control group. However, the fertilization rate and pregnancy rate were impaired by the contamination of endometriotic contents. We conclude that ovarian endometriosis should be treated before starting in vitro fertilization program in order to increase the pregnancy outcome.

Clinical study of a monophasic pill containing 20 microg ethinylestradiol and 150 microg desogestrel in Thai women.

OBJECTIVES: To evaluate the contraceptive reliability and clinical effects of a monophasic oral contraceptive containing 20 microg ethinylestradiol and 150 microg desogestrel (Mercilon) in Thai women. METHOD: The study was carried out at the Family Planning Clinic of King Chulalongkorn Memorial Hospital, Bangkok, Thailand. One hundred and forty six healthy women of fertile age were enrolled and treated with the study oral contraceptives for 12 cycles. Clinical data of vaginal bleeding, side effects, blood pressure and body weight were assessed periodically. RESULTS: One thousand four hundred and twenty five cycles were evaluated. No conception occurred. The cycles were almost regular. The incidence of irregular bleeding was highest in the first cycle, after the third cycle, the value was below 4 per cent. Side effects were very few. There was no change in blood pressure. A slight decrease in mean body weight was observed. CONCLUSION: The oral contraceptive containing 20 microg ethinylestradiol and 150 microg desogestrel has high contraceptive efficacy, good cycle control and minimal side effects.

Short course zidovudine maternal treatment in HIV-1 vertical transmission: randomized controlled multicenter trial.

A multicenter randomized, double blind, placebo-controlled clinical trial was conducted to evaluate the effectiveness of a short course of oral zidovudine (ZDV) treatment in HIV-1 infected pregnant women, starting at 38 weeks of gestation plus ZDV infusion during labor until delivery, to reduce HIV-1 vertical transmission in non-breast fed infants. One hundred and eighty two asymptomatic antiretroviral naïve HIV-1 infected pregnant women were enrolled. Each patient was randomly allocated into either the ZDV or placebo group. The ZDV group received 250 mg ZDV orally twice a day initiated at 38 weeks' gestation until the onset of labor. During the intrapartum period, ZDV infusion at the rate of 2 mg/kg was administered within the first hour and then continuously infused at the rate of 1 mg/kg/h until delivery. The placebo group received an identical capsule during pregnancy and normal saline infusion during labor until delivery. HIV-1 transmission was documented by nested polymerase chain reaction in infants at birth and at 1, 3 and, 6 months of age. The estimated HIV-1 vertical transmission rate was 14.9 per cent (95% CI = 11.1 to 18.7) and 16.3 per cent (95% CI = 12.3 to 20.9) in ZDV and placebo group, respectively (p > 0.05). The short course ZDV in antiretroviral naïve pregnant women initiated at 38 weeks' gestation plus intrapartum ZDV infusion without treatment in the infants was not effective to prevent HIV-1 vertical transmission.

Double uterus with unilaterally obstructed hemivagina and ipsilateral renal agenesis: a variety presentation and a 10-year review of the literature.

A double uterus with a unilaterally obstructed hemivagina is a rare condition, usually associated with ipsilateral renal agenesis. Herein, we report two cases, the first case presenting with abdominal pain and pelvic mass. Hemihysterectomy was performed leaving the contralateral uterus intact. The second case presented with chronic foul smelling vaginal discharge. The diagnosis was a double uterus and pyocolpos of the left vagina. Excision of the left vaginal septum and drainage were performed. The postoperative course of both cases was uneventful and the patients were well at the six-week follow-up. An accurate diagnosis, appropriate management and the prevention of future fertility problems are discussed.

Effects of different biopsy methods on the development of preimplantation mouse embryos, in vitro and in vivo: implication for preimplantation genetic diagnosis.

The aim of this study was to investigate the efficacy of two different embryo biopsy techniques, direct aspiration and partial zona dissection (PZD)-push, on subsequent in vitro and in vivo development of 8-cell stage mouse embryos. It was found that the rates of normal blastocyst formation and hatching blastocysts of direct aspiration, PZD-push, solution control and control embryos were not significantly different (80.8%, 81.6%, 84.5%, 86.7% and 71.9%, 72.3% and 74.6%) respectively. There was, however, a significant reduction in rate of complete hatching blastocysts (P < 0.1) (72.9% aspiration versus 85.2 per cent solution control and 86.4% control) and rate of live-born fetuses (24.2% aspiration versus 43.3% solution control and 41.2% control) (P < 0.05) in the direct aspiration group but no significant difference in the PZD-push group (80.3% of complete hatched blastocysts and 33.8% of live-born fetuses). These findings indicated that embryo biopsy with PZD-push was superior to the direct aspiration method. This mouse embryo biopsy model was useful in advancing development of biopsy technique for human preimplantation genetic diagnosis.

Clinical effects of gestrinone for the treatment of pelvic endometriosis in infertile patients.

The clinical efficacy and side effects of oral gestrinone, 2.5 mg twice weekly, were evaluated in this prospective study involving 22 patients with laparoscopically confirmed endometriosis. All patients came to the hospital with infertility problem. After 6 months of treatment, 81 per cent of patients had amenorrhea. Dysmenorrhea and pelvic pain were reduced from 90.9 per cent to 14.3 per cent and 81.8 per cent to 9.5 per cent respectively. Pelvic tenderness and induration were improved from 55.6 per cent and 50.0 per cent to 15.8 per cent and 10.5 per cent respectively. Androgenic effects such as acne was founded in 18.2 per cent of the patients. Return of fertility was observed in 25 per cent (5 patients) after 30-254 days post treatment. No serious side effect was detected during the treatment. The results suggest that gestrinone may be considered an option for the treatment of endometriosis related infertility.

Transvaginal hysterosalpingo-contrast sonography (HyCoSy) compared with chromolaparoscopy: a preliminary report.

Fifteen infertile women who required tubal passage evaluation by chromolaparoscopy were recruited. Those who had lower genital tract infection or abnormal uterine bleeding were excluded from the study. Transvaginal HyCoSy was performed during the first half of the menstrual cycle at least 24 hours prior to chromolaparoscopy. The results from both transvaginal HyCoSy and chromolaparoscopy were compared in assessing tubal patency and uterine pathology. A high correlation was noted regarding uterine examination using transvaginal HyCoSy compared with chromolaparoscopy (sensitivity, specificity, PPV and NPV were 91.7%, 33.3%, 84.6% and 50%, respectively). The correlation of the outcome between the two procedures in assessing tubal patency, when combining both tubes, was also high (sensitivity, specificity, PPV and NPV were 100%, 55.6%, 80% and 100%, respectively). The most common adverse event was only mild pelvic pain which did not necessitate any treatment. These preliminary results reveal the potential value of transvaginal HyCoSy as an alternative in infertility investigations. It seems to be as effective but less invasive than conventional chromolaparoscopy. The adverse events reported in this study relate more to the procedure (catheter insertion) rather than the trial substance.

Successful pregnancy in a case of azoospermia from congenital bilateral absence of vas deferens (CBAVD) using percutaneous epididymal sperm aspiration (PESA) and intracytoplasmic sperm injection (ICSI).

In men with obstructive azoospermia, bypass surgery would obviously be the most acceptable form of treatment as it gives the couple an opportunity to conceive naturally. However, when this has failed to restore patency or when surgery is not feasible (congenital absence of vas), fertility treatment using spermatozoa aspirated from the epididymis should be considered. Percutaneous epididymal sperm aspiration (PESA) is more acceptable to patients than micro epididymal sperm aspiration (MESA) because it eliminates the requirement for a general anaesthetic, post-operative pain, and the risk of haematoma formation, thus allowing a rapid return to normal activity of the husband. To our knowledge, this is the first reported case in Thailand where a pregnancy resulted in a couple whose infertility was due to azoospermia from the congenital absence of vas deferens. Spermatozoa collected through PESA were used in ICSI to achieve fertilization. The PESA technique, due to its simplicity is the choice of treatment for obstructive azoospermia.

Reliable sex determination of mouse preimplantation embryos by PCR amplification of male-specific genes in single blastomeres.

OBJECTIVE: To assess the reliability of sex determination in mouse preimplantation embryos using the two-step polymerase chain reaction method. SETTING: Division of Immunology, Department of Microbiology and Division of Reproductive Medicine, Department of OB/GYN. METHODS: The Sry and Zfy genes, known to be present in the sex-determining region of mouse Y chromosome, were selected for Y-specific target sequences and DXNds 3 locus located on mouse X chromosome was served as the internal control sequence. DNAs extracted from heart blood of male and female mice were used to test the correctness and specificity of the selected primers using the two-step PCR method. The same experimental conditions were then used to amplify the single copy genes in single mouse blastomeres with two pairs of primers for each of the target sequences. The sex-determined embryos were transferred to the uteri of pseudopregnant recipients to test the consistency of the assay system. RESULTS: All male and female blood DNA sample results confirmed the correct sex identification of the origin (100%). Nineteen of 20 single blastomeres showed the accurate diagnosis when compared with theirs 7/8 embryos. The sex of 36 of 37 mouse pups born from biopsied male and female embryos agreed with the predicted sex. CONCLUSION: The reliable genetic analysis of sex chromosome- specific sequences in single cell is possible by the two-step PCR method and could be applied for diagnosis of defective genes of human preimplantation embryos derived from the in vitro fertilization program.

Basal follicle--stimulating hormone levels on day 3 of previous cycle are predictive of in vitro fertilization outcome.

Serum FSH level may be an indicator of the functional potential of the ovary. The basic control of LH and FSH is by a negative feedback system involving the hypothalamus, anterior pituitary, and ovary. Basal serum FSH on day 3 of previous cycles in 36 patients underwent ovarian stimulation with a combination of GnRH agonist and FSH and hMG in an IVF program were evaluated. The results in this study show that a high basal serum FSH level is associated with a higher cancellation rate, a lower peak serum E2 level, lower number of oocytes retrieved and a lower clinical pregnancy rate.

Bilateral tubal pregnancy following in vitro fertilization and embryo transfer.

We have presented a case of bilateral tubal pregnancy following in vitro fertilization and embryo transfer. The most likely cause, in our opinion, is 'spray effect' which was discussed. The diagnosis of ectopic pregnancy must always be kept in mind in IVF&ET program, due to its high incidence of occurrence compared to natural conception. Although the incidence of bilateral tubal pregnancy is not high, one should identify the opposite adnexa when tubal pregnancy is diagnosed in an IVF&ET patient. Early diagnosis is essential for prevention of significant maternal morbidity and mortality.

Growth and development of infants conceived by in vitro fertilization: a preliminary report.

This is a preliminary report on the first 24 babies born during the period of 1 year follow-up after IVF-ET in Chulalongkorn Hospital. Based on this study, our data suggests that these children have a normal range of growth and development. There was also no evidence of major congenital anomalies identified in spite of some minor ones. However, a need for more detail and on-going long-term follow-up at least until school age is being planned.