Variable aneuploidy mechanisms in embryos from couples with poor reproductive histories undergoing preimplantation genetic screening.

BACKGROUND: Preimplantation genetic screening (PGS) is used to determine the chromosome status of human embryos from patients with advanced maternal age (AMA), recurrent miscarriage (RM) or repeated implantation failure (RIF). METHODS: Embryos from 47 such couples were investigated for chromosomes 13, 15, 16, 18, 21 and 22 using fluorescence in situ hybridization with two rounds of hybridization. The investigation included parental lymphocyte work-up, the screening of blastomeres on day 3 and full follow-up on day 5/6 of untransferred embryos. RESULTS: The outcome of 60 PGS cycles is described, in which 523 embryos were biopsied; 91% gave results, of which 18% were diploid for all the chromosomes tested and 82% were abnormal. The pregnancy rate per cycle that reached the biopsy stage was 27%, and 30% per embryo transfer. Satisfactory follow-up was obtained from 353 embryos; all those diagnosed as abnormal were confirmed as such, although two false-positives were detected in relation to specific chromosome abnormalities. Meiotic errors were identified in 16% of embryos. Between the RM, AMA and RIF groups, there was a significant difference in the distribution of embryos that were uniformly abnormal and of those with meiotic errors; with an almost 3-fold increase in meiotic errors in the first two groups compared with the RIF group. CONCLUSIONS: This complete investigation has identified significant differences between referral groups concerning the origin of aneuploidy in their embryos.

Antral follicle count, anti-mullerian hormone and inhibin B: predictors of ovarian response in assisted reproductive technology?

OBJECTIVE: The objective of this study was to evaluate the relationship between anti-mullerian hormone (AMH), inhibin B and antral follicle count (AFC) with ovarian response. DESIGN: Retrospective study. SETTING: Fertility unit. SAMPLE: AFC was recorded, and a serum sample obtained on day 3 from all patients undergoing in vitro fertilisation (IVF). Patients were given 300 IU/L recombinant follicle stimulating hormone (FSH; Gonal F). The following day blood samples were collected. METHODS Serum samples were assayed for FSH, AMH and inhibin B using commercial immunoassay kits and oestradiol using an in house assay. MAIN OUTCOME MEASURES: Response to gonadotrophin stimulation and the number of eggs collected. RESULTS: AFC was negatively correlated to age (r=-0.426, P < 0.001). Delta inhibin B (levels of inhibin B on day 4 minus day 3) had the best association to the number of eggs collected (r= 0.533, P < 0.001) followed by basal AMH (r= 0.51, P < 0.001) and AFC (r= 0.505, P < 0.001). The number of eggs fertilised was significantly associated with basal AMH (r= 0.592, P < 0.001) and inhibin B (r= 0.548, P < 0.001). AMH with a cutoff of 0.2 ng/mL had the best sensitivity (87%) and specificity (64%) in predicting poor response. A cumulative score using basal FSH, basal AMH, delta E2 (levels of oestradiol on day 4 minus day 3), delta inhibin B, AFC and age gives the best predictive statistics to identify poor responders with 87% sensitivity and 80% specificity and a positive likelihood ratio of 4.36. CONCLUSION: Delta inhibin B had the best positive association with the number of eggs collected and basal AMH is the single best predictor of poor response. AFC has a significant association with the number of eggs collected and is predictive of clinical pregnancy. It is evident that a single parameter is of limited value in predicting ovarian response. However, we have demonstrated a cumulative score using all the above markers could be useful in predicting poor response.

Preimplantation genetic diagnosis of chromosome abnormalities: implications from the outcome for couples with chromosomal rearrangements.

OBJECTIVES: Chromosomal rearrangements can lead to infertility or repeated spontaneous or induced abortions. The use of preimplantation genetic diagnosis (PGD) allows the selected transfer of chromosomally balanced embryos. The aim of this study was to carry out detailed analysis of the outcome of 11 PGD cycles for 8 patients carrying various chromosomal rearrangements. METHODS: Patients underwent routine in vitro fertilisation with biopsy of embryos on day 3. Specific fluorescent in situ hybridisation protocols were developed for each couple. Embryo transfer was possible in all 11 cycles. RESULTS: The outcome was four pregnancies, leading to three live births and one biochemical pregnancy. Post-zygotic mosaicism was detected in 75% of untransferred embryos, the majority of which were chaotic. Detailed follow-up and analysis provided evidence for the co-existence of chromosomally balanced and abnormal cells in six embryos. The mechanisms involved included chromosome breakage and loss of material. CONCLUSIONS: Biopsy and analysis of two blastomeres, where possible, reduced the risk of misdiagnosis in cases of balanced/aneuploid mosaics. The three live births achieved for the eight couples treated in this series, despite the poor history in almost all cases, is further proof that a policy of biopsying two cells from embryos consisting of six or more cells and a single cell from four- or five-cell embryos is compatible with a positive outcome.

Simultaneous evaluation of basal FSH and oestradiol response to GnRH analogue (F-G-test) allows effective drug regimen selection for IVF.

To determine whether preliminary assessment of ovarian reserve by simultaneous evaluation of basal follicle-stimulating hormone (FSH) and oestradiol response to gonadotrophin releasing hormone (GnRH) analogue (F-G-test) can be used to tailor individually the drug regimen for ovarian stimulation, the in-vitro fertilization (IVF) results of 238 patients were retrospectively analysed. Sixty-two women with abnormal response to the test (DeltaE2 <180 pmol/l and/or FSH >9.5 mIU/ml) had commenced buserelin nasal spray in the mid-luteal phase and discontinued it on cycle day 1. Ovarian stimulation was started on cycle day 3 with 375 IU/day of gonadotrophin. Fifty-three patients completed the treatment cycle (group A). A total of 176 women with normal response to the test (DeltaE2 >180 pmol/l and FSH <9.5 mIU/ml) had continued the GnRH analogue throughout the stimulation cycle and a starting dose of 225 IU/day of gonadotrophin was used from cycle day 3. A total of 158 patients completed the treatment cycle (group B). Group A had significantly higher age (34.9 +/- 4.2 versus 33.2 +/- 4.2) (P < 0.05) and basal FSH (9.2 +/- 3.8 versus 7.0 +/- 2.2) (P < 0.05) and required a higher total dose of gonadotrophin. The numbers of oocytes retrieved and embryos transferred were significantly lower. However, fertilization, clinical pregnancies, and implantation rates were similar in both groups. It was concluded that simultaneous evaluation of basal FSH and oestradiol response to GnRH analogue can be useful in identifying subcategories of women with reduced ovarian reserve who may benefit from reduced GnRH analogue administration and a higher starting dose of gonadotrophin.

Basal 17beta-estradiol did not correlate with ovarian response and in vitro fertilization treatment outcome.

OBJECTIVE: To verify the correlation of basal 17beta-E(2) with ovarian response to stimulation and outcome of in vitro fertilization (IVF). DESIGN: Retrospective observational study. SETTING: The Assisted Conception Unit, University College London Hospitals. PATIENT(S): Three hundred five women undergoing IVF and IVF with intracytoplasmic sperm injection. INTERVENTION(S): Basal follicle-stimulating hormone (FSH) and 17beta-E(2) were assessed. The cutoff level for day 2 E(2) established was 250 pmol/L. Each patient was noted for below (group A) or above (group B) the cutoff point according to her basal E(2) level. MAIN OUTCOME MEASURE(S): Basal E(2), age, duration of infertility, cycle day 2 FSH, number of ampules of gonadotropin used, number of days of stimulation, number of retrieved oocytes, fertilization rate, number of embryos transferred, number of cycles with embryo freezing, cancellation rate, clinical pregnancy rate, and implantation rate were compared between the two groups. RESULT(S): No differences were found between group A and group B in the number of oocytes retrieved (8.8 +/- 4.2 vs. 9.3 +/- 4.8), embryos transferred (2.5 +/- 0.8 vs. 2.7 +/- 0.7), cancellation (9.1% vs. 6.9%), pregnancy (24.8% vs. 30%), and implantation rate (12.3% vs. 15.6%). Correlation coefficient and coefficient of determination showed no significant correlation between basal E(2) and the number of oocytes retrieved, age, and basal FSH. CONCLUSION(S): In our study population, basal E(2) was not a sensitive predictor of ovarian response to stimulation and did not correlate with IVF outcome.

Simultaneous evaluation of basal follicle-stimulating hormone and 17 beta-estradiol response to gonadotropin-releasing hormone analogue stimulation: an improved predictor of ovarian reserve.

OBJECTIVE: To compare different predictors of ovarian reserve. DESIGN: Prospective study. SETTING: The Assisted Conception Unit, University College London Hospitals. PATIENT(S): One hundred seventy-seven patients undergoing IVF treatment. INTERVENTION(S): Blood samples were collected on cycle day 2 to determine basal levels of FSH and 17beta-E2 and the FSH/LH ratio, and on cycle days 3 and 4 to assess the increase in FSH (deltaFSH) and 17beta-E2 (deltaE2) after the commencement of GnRH analogue (GnRH-a) stimulation. Ultrasound scans were performed during ovarian stimulation to assess the number of follicles. MAIN OUTCOME MEASURE(S): Day 2 FSH and 17beta-E2 levels, the FSH/LH ratio, and the deltaFSH and deltaE2 after the commencement of GnRH-a stimulation were correlated with the number of follicles obtained after ovarian stimulation. RESULT(S): All the possible predictors considered, except for the day 2 E2 level and the deltaFSH, correlated significantly with the ovarian response. The best single correlation was between the number of follicles and the deltaE2 (GnRH-a test). When the FSH level was evaluated simultaneously, the correlation was strengthened, resulting in a better negative predictive value. CONCLUSION(S): Simultaneous evaluation of basal levels of FSH and of the response of E2 to GnRH-a stimulation seems to be the best marker of ovarian reserve and a sensitive predictor of response to ovarian stimulation in patients undergoing IVF treatment.

Oocyte morphology predicts outcome of intracytoplasmic sperm injection.

To examine the influence of cytoplasmic morphology on the success rate of intracytoplasmic sperm injection (ICSI), the morphology of 837 metaphase II oocytes was assessed after cumulus stripping. The main abnormalities detected were excessive granularity, cytoplasmic inclusions such as vacuoles, smooth endoplasmic reticulum clustering and refractile bodies. Microinjection was performed in 538 oocytes with normal cytoplasm, 142 out of 161 with excessive granularity and 112 out of 138 with cytoplasmic inclusions. Very poor oocytes were not injected. No difference was found in fertilization rate. The embryos achieved cleaved normally and a similar number of good quality embryos among the three groups was noted. The outcome of transfer of embryos derived solely from normal oocytes (group A: 72 patients, 183 embryos) was compared with those from oocytes with cytoplasmic abnormalities (group B: 34 patients, 85 embryos). In group A, 17 clinical pregnancies (24% per patient, implantation rate 10%) were established. In group B, only one clinical pregnancy (3% per patient, implantation rate 1%) was established, from the transfer of embryos derived from oocytes with homogeneous granularity of the cytoplasm. No pregnancy resulted following the transfer of embryos from eggs with cytoplasmic inclusions. The difference was statistically significant. The outcome of ICSI is dependent on the quality of the oocytes retrieved. Normal fertilization and early embryo development were achieved in oocytes with abnormal cytoplasm morphology, but the resulting embryos failed to demonstrate the same implantation potential as those derived from oocytes with normal cytoplasm.

Successful establishment of pregnancy by superovulation and intrauterine insemination with sperm recovered by a modified Hotchkiss procedure from a patient with retrograde ejaculation.

OBJECTIVE: To improve the quality of the sperm recovered from the bladder in a patient with retrograde ejaculation who already had failed to conceive after several attempts at IUI with sperm recovered by conventional techniques. SETTING: University Hospital. PATIENTS: A couple with male infertility due to retrograde ejaculation caused by the Zielke operation, a spinal fixation procedure performed to correct severe kyphoscoliosis. INTERVENTION: Superovulation and IUI of sperm recovered from the bladder using a modified Hotchkiss procedure involving the introduction into the bladder of Earle's balanced salt solution (EBSS) buffered with Hepes in sufficient quantity to bring the urinary pH and osmolarity to those of fresh ejaculate. MAIN OUTCOME MEASURES: Urine pH and osmolarity at baseline and after dilution with EBSS buffered with HEPES. Concentration, motility, and progression score of the sperm recovered from the bladder. RESULTS: Good sperm samples were achieved. Pregnancy was established when IUI was performed in association with superovulation induction. CONCLUSIONS: Determination of urine pH and osmolarity appears to be a useful method for choosing the ideal sperm recovery procedure. The modified Hotchkiss procedure described seems to be a promising alternative method for recovering sperm for artificial insemination.

[Heterotopic ovarian pregnancy after in-vitro fertilization and embryo transfer and contralateral tubal pregnancy after gamete intrafallopian transfer]. / Gravidanza eterotopica ovarica dopo fertilizzazione in vitro ed embryo transfer e gravidanza tubarica controlaterale dopo trasferimento intratubarico dei gameti.

One case of ovarian ectopic pregnancy associated with twin intrauterine viable pregnancy after IVF-ET, and one case of contralateral tubal ectopic pregnancy after GIFT performed with normal fallopian tubes, are reported. The possible pathogenesis and management are discussed.

Ovarian heterotopic pregnancy after IVF and contralateral tubal ectopic pregnancy after GIFT.

One case of ovarian ectopic associated with twin intrauterine viable pregnancy after in vitro fertilization, and one case of contralateral tubal ectopic pregnancy after gamete intra fallopian transfer performed with normal fallopian tubes, are reported. The possible pathogenesis and the management are discussed. The risk of ectopic pregnancy after assisted conception can be present even when the fallopian tubes are apparently normal.

Twin pregnancy complicated by intrauterine demise of one fetus.

Two cases of antepartum demise of one fetus in a twin pregnancy are reported. Obstetrical management and fetal outcome are discussed with particular respect to changes in maternal coagulation parameters.