Jane L. Frederick

Preovulatory follicular fluid steroid levels in stimulated and unstimulated cycles triggered with human chorionic gonadotropin.

The purpose of this study was to analyze follicular fluid (FF) samples for steroid levels from stimulated and unstimulated cycles triggered with human chorionic gonadotropin (hCG) and to assess the influence of controlled ovarian hyperstimulation and luteinizing hormone/hCG on these levels. Spontaneous ovulatory cycles were monitored with serial ultrasound examinations, and hCG 10,000 IU was given when the lead follicle was mature. Fourteen FF samples yielding fertilizable oocytes were compared with 13 FF samples from controlled ovarian hyperstimulation cycles. Progesterone (P) was higher in controlled ovarian hyperstimulation than in unstimulated cycles (9.0 +/- 1.2 micrograms/mL versus 4.4 +/- 0.6 microgram/mL; mean +/- SEM), whereas estradiol (E2) was lower (0.8 +/- 0.1 microgram/mL versus 1.3 +/- 0.2 microgram/mL), resulting in a higher P:E2 ratio (15.5 +/- 3.3 versus 4.4 +/- 0.7). Androstenedione (A), testosterone (T), and T:E2 ratios were all higher in unstimulated than controlled ovarian hyperstimulation cycles. We conclude that controlled ovarian hyperstimulation is associated with increased FF P, decreased FF E2, T, and A levels, and decreased T:E2 ratios, suggesting altered steroidogenesis and enhanced follicular aromatase activity.

Initiation of angiogenesis by porcine follicular fluid

Angiogenesis was observed and measured after injection of porcine follicular fluid into rabbit corneas. A qualitative response (0 to 6+) and quantitative measurement (mm/day) were obtained 9 days after injection. Undiluted porcine follicular fluid stimulated angiogenesis with new blood vessels visible by the third day after injection, extending 2.0 to 3.0 mm into the site of injection from the corneal scleral limbus (1 to 4+) by day 9. Angiogenic activity was consistently found in fractions of porcine follicular fluid which precipitated in 20% to 40% saturated ammonium sulfate. Sephadex gel filtration of the 20% to 40% saturated ammonium sulfate fraction resulted in fractions with molecular weights of 45,000 to 60,000 and less than or equal to 1500 daltons which stimulated angiogenesis. Charcoal treatment of active fractions did not remove angiogenic activity. Angiogenic activity was retained after heating at 56 degrees C for 1 hour but was lost after boiling (20 minutes). Quantitative measurements of chemotaxis with use of Boyden chambers and mitogenesis by means of tritiated thymidine incorporation were performed. Follicular fluid from small follicles contained greater chemotactic activity than follicular fluid from medium or large follicles. The 20% to 40% saturated ammonium sulfate precipitate that eluted through Sephadex G-100 with a molecular weight of 45,000 to 60,000 daltons contained angiogenic, mitogenic, and chemotactic activity. In conclusion, porcine follicular fluid contains angiogenic factors that may be associated with perifollicular neovascularization during folliculogenesis.

The effect of leuprolide acetate on steroidogenesis by granulosa and theca cells in vitro

Enhancement of follicular response to controlled ovarian hyperstimulation for human in vitro fertilization (IVF) has been suggested following pretreatment with leuprolide acetate (LA). However, additional human menopausal gonadotropin (hMG) is required to achieve follicle maturity in the presence of LA. We studied the effect of LA on steroidogenesis of granulosa and theca cells in vitro. Human granulosa cells obtained from IVF follicular fluid aspirations were cultured for 14 days in the presence and absence of human chorionic gonadotropin (hCG). hCG significantly enhanced progesterone (P) and estradiol (E2) production by the cells, however, the addition of LA in concentrations of 10, 100, and 1000 ng/ml had no effect. Porcine granulosa cells were cultured for 48 hr in the presence and absence of follicle stimulating hormone (FSH) with the addition of LA at the same doses. LA did not affect the FSH-induced increase in P production. Porcine theca cells were cultured for 48 hr in the presence and absence of hCG. The addition of LA did not affect androstenedione (A) production by these cells. We conclude that in this dynamic model in vitro, LA does not inhibit or stimulate P or E2 production by granulosa or A production by theca cells.

Effect of hydration on random levels of urinary pregnanediol glucuronide.

To determine the effect hydration has upon the relationship between serum progesterone and its urinary metabolite pregnanediol-3 alpha-glucuronide (PDG) we measured spot samples from patients (n = 207) presenting in the 1st trimester of pregnancy. Serum and urine specimens were obtained simultaneously and measured for progesterone and PDG by radioimmunoassay (RIA). Urine specific gravity was also measured at the time of sample collection. Results demonstrated that in samples where the urine specific gravity was greater than or equal to 1.015 there was a strong positive correlation between serum progesterone and PDG (r greater than 0.60, p less than 0.001). However, when urine samples were below a specific gravity of 1.015, serum and urinary steroid values correlated poorly or not at all. We conclude that the measurement of specific gravity should be routinely performed when determining random values of urinary PDG, since only samples in which the urine is adequately concentrated accurately reflect corresponding serum progesterone concentrations.

Is There a Role for Ovarian Stimulation and Intra-Uterine Insemination After Age 40?

The objective of this study was to determine the conception rate in infertile couples in which the female partner was > or = 40 years old and who had received ovarian stimulation treatment and intra-uterine insemination (IUI). It was a retrospective study of 77 patients who underwent a total of 210 treatment cycles. Protocols for ovulation induction included clomiphene citrate, human menopausal gonadotrophin (HMG) and clomiphene citrate plus HMG. Patients were monitored using transvaginal ultrasound, and two IUI were performed 24 and 48 h after the determination of urinary luteinizing hormone (LH) surge or human chorionic gonadotrophin (HCG) injection. A total of 11 pregnancies were reported, giving a pregnancy rate of 14% per patient and 5% per cycle. Eight spontaneous abortions occurred, giving a pregnancy wastage of 73%. In a previous comparative analysis of 543 patients < 39 years old receiving IUI and identical protocols of ovarian stimulation, 141 pregnancies were achieved, giving a pregnancy rate of 21% per patient and 10% per cycle. The miscarriage rate in that group was 18%. This report compares IUI results for women > or = 40 years with those obtained previously for younger women, and shows the very poor success rate in women > 40 years of age. This information will be important in the proper counselling of this group of patients, as well as indicating that a prompt recommendation for assisted reproductive treatment should be made soon after the failure of a few attempted cycles of ovarian stimulation treatment and IUI.

Frozen Zygote Intrafallopian Transfer: A Successful Approach for Transfer of Cryopreserved Embryos

OBJECTIVE To assess whether frozen ET to the fallopian tube is a possible alternative for cryopreserved embryos. DESIGN Fifty-four patients (mean age 35 years) participated, in which their embryos were cryopreserved in 1.5 M propanediol at the pronuclear or two-cell stage. Each patient then underwent a steroid replacement cycle consisting of oral micronized 17 beta-E2 2 mg on days 2 to 4, 4 mg on days 5 to 7, 6 mg on days 8 to 10, and 8 mg from day 11 on. Serial ultrasounds were performed to evaluate the endometrium until an optimal thickness of > or = 10 mm triple layer was achieved. At this time, 100 mg IM progesterone was initiated and the zygote intrafallopian transfer (ZIFT) procedure was performed on the third day of P administration. The average number of embryos transferred was 4.4. RESULTS Twenty-two clinical pregnancies resulted, giving a pregnancy rate of 41%. Eight miscarriages occurred and one ectopic pregnancy resulted, giving a live birth rate of 24%. Implantation rate was 10.8%. The highest chance of pregnancy was seen in patients who never had a previous IVF, GIFT, or ZIFT (61%). CONCLUSION This is the first report of a series of frozen ETs to the fallopian tubes. These results indicate that tubal transfer may offer a protective benefit of the tubal environment and avoidance of endometrial trauma, and should be added to our armamentarium of replacing cryopreserved embryos.