Protein phospholipase C Zeta1 expression in patients with failed ICSI but with normal sperm parameters.

PURPOSE: This study was conducted to determine if expression of the testis-specific phospholipase C Zeta1 (PLCZ1) correlated with low success or fertilization failure after ICSI in patients with normal parameters after standard semen analysis (SA). METHODS: Couples <43 years with one or two failed or low fertilization ICSI cycles. Standard Semen Analysis (SA) was performed to determine sperm parameters in male partners, whereas females were evaluated for antral follicle counts (AFC), day 3 FSH levels and peak Estradiol (E2) levels. The presence of PLCZ1 in sperm was ascertained using Western blotting and Immunofluorescence (IF) analysis. The ability of sperm to initiate changes in the intracellular concentrations of free calcium ([Ca(2+)]i), which is characteristic of mammalian sperm, was performed after injection of human sperm into mouse eggs loaded with the Ca(2+) sensitive dye fura-2 AM. RESULTS: Male partners of couples with failed or low success ICSI fertilization but with normal SA parameters showed low expression levels of PLCZ1 as determined by western blotting and reduced fluorescent signal during IF studies. In addition, fewer of these males' sperm showed PLCZ1 expression and were able to initiate robust [Ca(2+)]i oscillations upon injection into eggs. CONCLUSION: Our data suggest that in patients with normal SA parameters but with repeated low fertilization or outright failed fertilization results after ICSI, abnormal PLCZ1 function should be considered as the underlying mechanism responsible for the failure of fertilization.

Influence of embryo transfer depth on in vitro fertilization and embryo transfer outcomes.

OBJECTIVE: To investigate the influence of transfer distance from the fundus (TDF) on clinical pregnancy rate (PR) and ectopic pregnancy rate. DESIGN: Retrospective cohort. Between January 2000 and December 2001, 699 ultrasound (US)-guided embryo transfers were conducted. Mock transfer was performed to measure uterine cavity depth 1 month before treatment. Cavity depth was measured by abdominal US before the transfer, from the vaginal stripe to the fundus. Transfers were performed with a Wallace embryo transfer catheter (Cooper Surgical, Shelton, CT) using US and physician's judgment of cavity depth. Transfer distance from the fundus was calculated by subtracting the depth of catheter insertion from the cavity depth, as determined by US or by mock transfer. Statistical analyses were performed by building a multivariable logistic regression model to calculate odds ratios and 95% confidence intervals (CI). SETTING: Women aged 23 to 43 years who are in a university-affiliated, community-based IVF program in Springfield, Massachusetts. PATIENT(S): All patients enrolled in IVF program undergoing embryo transfer. INTERVENTION(S): No patient received any additional procedure or intervention. All of the measurements obtained with the embryo catheter and the transvaginal ultrasound were part of the program's protocol for the embryo transfer. MAIN OUTCOME MEASURE(S): Odds ratio examining relationship between embryo transfer depth and PR.Clinical, implantation, and ectopic PR were 37%, 20%, and 2.1%. Cavity depth by US differed from cavity depth by mock by at least 10 mm in >30% of cases. The TDF by US was highly predictive of PR; TDF by mock was not predictive of PR. Increasing the TDF by US resulted in significantly increased PR as well as lower ectopic rates. Using regression analysis, the odds ratio for TDF by US was 1.11 (95% CI: 1.07-1.14). This suggests that for every additional millimeter embryos are deposited away from the fundus, the odds of clinical pregnancy increased by 11%. CONCLUSION(S): After controlling for potential confounders, the clinical PR is significantly influenced by the transfer distance from the fundus. Cavity depth by US is clinically useful to determine the depth beyond which catheter insertion should not occur.

Antral follicle count in clinical practice: analyzing clinical relevance.

OBJECTIVE: To determine the clinical relevance of obtaining antral follicle counts (AFC) before ovarian stimulation in an IVF program. DESIGN: Retrospective cohort study. SETTING: An IVF program in a large academic teaching hospital. PATIENT(S): A total of 1,049 stimulated IVF cycles in 734 subjects between September 2003 and December 2007 selected from our program's database. INTERVENTION(S): Basal antral follicles (AFCs) (3 mm-10 mm) were counted via ultrasound scan on cycle day 3 in luteal leuprolide acetate stimulations, or after at least 2 weeks of oral contraceptives in microdose leuprolide acetate stimulations. Patients were grouped according to basal AFC, and outcome parameters compared for AFC groups within each stimulation protocol. MAIN OUTCOME MEASURE(S): Oocytes retrieved, ovarian response, implantation rate, cancellations, pregnancy, pregnancy loss, and live births per cycle start. RESULT(S): Antral follicle count grouping is predictive of threefold change in ovarian response to gonadotropins and oocytes retrieved. Low AFC did predict a higher cancellation rate. Antral follicle count did not predict implantation rate, pregnancy rate, or live birth rate per cycle start. CONCLUSION(S): Antral follicle count may be helpful in determining stimulation protocol, as it is the most reliable determinant of oocytes retrieved per starting FSH dose. Antral follicle count predicts ovarian response, not embryo quality or pregnancy.

Turner mosaicism (45,X/46,XX) diagnosed in a young woman subsequent to low oocyte maturity and failed ICSI.

OBJECTIVE: To report a case of Turner mosaicism with low oocyte maturity during ICSI. DESIGN: Retrospective case report. SETTING: Academic tertiary medical center. PATIENT(S): A 28-year-old patient with 7-year history of primary unexplained infertility. INTERVENTION(S): Four cycles of ICSI. MAIN OUTCOME MEASURE(S): Oocyte quality; fertilization, cleavage, and implantation rates; and live births during ICSI cycles; as well as patient karyotype were obtained. RESULT(S): Over four cycles of ICSI, 111 oocytes were harvested, a high follicular response, but 68% were immature (germinal vesicles or metaphase I). Additionally, seven embryos were transfered, but none implanted. Owing to this unusual response in a young healthy woman, maternal karyotype was performed, which demonstrated 17% Turner mosaicism. CONCLUSION(S): This is, to our knowledge, the first reported case of excellent ovarian response but marked oocyte immaturity in a young woman with Turner mosaicism. The implications of sex chromosome aneuploidy on fertility and the significance of low oocyte maturity are discussed.

Does catheter type effect pregnancy rate in intrauterine insemination cycles?

PURPOSE: To determine if a soft insemination catheter increases pregnancy rates. METHODS: Seven hundred forty-seven consecutive intrauterine insemination (IUI) cycles in 364 women in a reproductive endocrinology office between October 1998 and March 2000. Patients with even birth years were inseminated with the Tomcat catheter, and those with odd birth year with the Wallace IUI catheter. Clinical pregnancy rates were compared. RESULTS: The Wallace catheter group included 180 women for 372 cycles and an overall pregnancy rate of 16.4%. The Tomcat catheter group included 184 women for 375 cycles and an overall pregnancy rate of 18.1%. This difference is not statistically significant (P = 0.61). Potential confounders were accounted for. CONCLUSIONS: When comparing the softer Wallace catheter to the less pliable Tomcat catheter during IUI cycles, there was no significant difference in pregnancy rate when using a standard gentle technique that includes not touching the top of the fundus with the catheter.