STUDY QUESTION:Is there an optimal time to perform ICSI with respect to the times of oocyte pick-up (OPU), in order to maximize the reproductive outcomes in cycles with fresh and vitrified/warmed donor oocytes?SUMMARY ANSWER:We found no significant differences in reproductive outcomes of ICSI cycles within a wide range of times between OPU and ICSI.WHAT IS KNOWN ALREADY:In assisted reproduction, the oocyte is subject to denudation, vitrification/warming and ICSI. As shorter interaction with cumulus cells, oocyte ageing in vitro and insufficient recovery after warming may all impact the resulting embryo developmental competence, strictly controlled times between procedures are often implemented. However, most protocols have not been tested with the aim to improve reproductive results, and little information is available on the ideal times to be followed during these steps in order to optimize fertilization rates and embryo quality, and to achieve the highest pregnancy rate.STUDY DESIGN, SIZE, DURATION:Data from 3986 ICSI cycles performed between December 2012 and May 2014 were included (3178 with fresh and 808 with vitrified/warmed donor oocytes).PARTICIPANTS/MATERIALS, SETTING, METHODS:ICSI was performed using donor oocytes and either partner or donor sperm. Exact times between OPU, denudation, vitrification, warming and ICSI were recorded automatically by a radiofrequency-based system. OPU was performed strictly 36 h after GnRH agonist trigger. Biochemical pregnancy was defined as a positive serum βHCG 15 days after transfer, clinical pregnancy was defined as a visible embryo with heartbeat 5 weeks after transfer, and ongoing pregnancy was defined as a normally developing pregnancy at 12 weeks after transfer.MAIN RESULTS AND THE ROLE OF CHANCE:Times between OPU and ICSI (OPU-ICSI) ranged from 1 h 25 min to 17 h 13 min (averagefresh ± SD = 4 h 58 m ± 1 h; averagevitrified= 9 h 18 m ± 2 h). We found no effect of OPU-ICSI time on fertilization rate (pfresh=0.39; pvitrified=0.86) or embryo quality at Days 2 and 3 (pfresh=0.08; pvitrified=0.22). There was no difference in average OPU-ICSI times between positive and negative pregnancies (biochemical, clinical, ongoing and live birth rates) in either fresh (P = 0.71, 0.43, 0.79, 0.96) or vitrified (P = 0.59, 0.33, 0.73, 0.87) oocytes, respectively. Data were adjusted for oocyte donor age, semen status, number of motile spermatozoa and sperm concentration, and no effect of OPU-ICSI time on pregnancy and live birth rates for either fresh (P = 0.57, 0.16, 0.11, 0.46) or vitrified (P = 0.80, 0.73, 0.91, 0.95) oocytes was found. Further analysis for linear trend using OPU-ICSI time categorized in deciles showed that pregnancy rates and live birth rates do not increase or decrease across deciles. We found no effect of time taken for denudation to vitrification, warming to ICSI and denudation to ICSI on pregnancy rates.LIMITATIONS, REASONS FOR CAUTION:This is a study with automatically collected times from a high number of ICSI cases; however, its retrospective nature cannot exclude the influence of unaccounted for variables on the results. All oocytes came from oocyte donors (≤35 years old), so results cannot be extended to older or infertile women.WIDER IMPLICATIONS OF THE FINDINGS:Our results indicate that the effective window of time for insemination by ICSI might be wider than previously thought. It therefore appears that, within appropriate time frames, the management of ICSI cycles involving oocytes from young women in embryology laboratories could be adjusted to accommodate caseloads and workflow with no loss of oocyte viability or cycle efficiency.

STUDY QUESTION: Does the time from ovum pick-up (OPU) to frozen embryo transfer (FET) affect reproductive outcomes in a freeze-all strategy?SUMMARY ANSWER: Our study did not detect statistically significant differences between first and subsequent cycles, clinically relevant differences are not ruled out and further and larger studies are required.WHAT IS KNOWN ALREADY: Following controlled ovarian hyperstimulation (COH) delaying FET until the endometrium has returned to an optimal pre-stimulation state may have a significant emotional impact on patients, which adds to the stress and anxiety accompanying a standard IVF cycle. Currently there is no agreement on the best time to perform a FET after a freeze-all cycle in order to maximize reproductive outcomes for the patient.STUDY DESIGN, SIZE, DURATION: Retrospective cohort study of 512 freeze-all cycles, performed between January 2012 and December 2014. COH was performed by either a GnRH antagonist (n = 397) or a long GnRH agonist protocol (n = 115). Ovulation was triggered using either a GnRH agonist (n = 258) or hCG (n = 254). Endometrial preparation was performed in an artificial cycle by either oral (n = 238) or transdermal (n = 274) oestrogen. Differences were considered significant if P < 0.05.PARTICIPANTS/MATERIALS, SETTING, METHODS: Reproductive outcomes between FETs which took place either within the first menstrual cycle following OPU (Cycle 1; n = 263) or afterwards (Cycle = 2; n = 249) were compared. Student's t-test for independent samples, Mann-Whitney U-test and Chi-square analysis were used where appropriate. A multivariable logistic regression analysis was performed adjusting for maternal age, drug used for ovulation trigger, number of retrieved oocytes, number of embryos obtained, day of embryonic development at transfer, number of embryos transferred and type of endometrial preparation. Differences were considered significant if P < 0.05.MAIN RESULTS AND THE ROLE OF CHANCE: Live birth rate (LBR) was significantly higher in FET performed during Cycle 1 vs Cycle >= 2 (37.6% vs 27.3%, respectively; P = 0.01) before adjusting for confounding factors. We found no difference for biochemical pregnancy (49.8% vs 43.8%; P = 0.17), clinical pregnancy (44.1% vs 36.1%; P = 0.07) or pregnancy loss (11.8% vs 16.1%; P = 0.16). A multivariable analysis found no impact of timing of elective FET on LBR (odds ratio, OR 0.73; 95% CI 0.49-1.08). The impact remained not significant after adjusting for number of retrieved oocytes, drug used for ovulation trigger (hCG vs GnRH agonist) and reason for cryopreservation. The factors that significantly affected LBR were: maternal age in both age categories (women between 35 and 40 years vs women below 35 years, OR 0.63, 95% CI 0.4-0.95; and women over 40 years vs women below 35 years, OR 0.34, 95% CI 0.2-0.7), day of embryonic development at transfer (day +4 vs +3; OR 1.7, 95% CI 1.1-2.8) and number of transferred embryos (OR 2.2, 95% CI 1.4-3.3) and oestrogen used for endometrial preparation (transdermal vs oral; OR 0.62, 95% CI 0.4-0.9).LIMITATIONS REASONS FOR CAUTION: The main limitation of our study is its retrospective nature. Although we adjusted our statistical analysis for a number of known and suspected confounders, we cannot exclude the possibility of residual confounding factors.WIDER IMPLICATIONS OF THE FINDINGS: According to our results, clinicians might not need to wait more than one menstrual cycle before performing FET. This allows us to reduce unnecessary delays in FET, without compromising reproductive outcomes.

STUDY QUESTION:Is there a difference in live birth rates following endometrial preparation with either a constant or increasing estrogen dose in fresh embryo transfer from oocyte donation cycles?SUMMARY ANSWER:There is no difference in live birth rates between a constant dose versus an increasing dose of estrogen after fresh embryo transfer in oocyte donation cycles with oral or transdermal supplementation.WHAT IS KNOWN ALREADY:Endometrial preparation (EP) with estrogen and progesterone, and embryo-endometrial synchronicity are determinant for adequate embryo implantation. Estrogen is crucial and different exogenous administration patterns could imply variations on EP. Moreover, estrogen undergoes metabolization by the intestines and liver when administered orally, an effect that is bypassed by transdermal administration. Information on the effect of replacement patterns and route of administration of E on reproductive outcomes of women undergoing fresh embryo transfer from oocyte donation cycles is scarce.STUDY DESIGN, SIZE, DURATION:Retrospective cohort study including 8362 embryo transfers following ICSI, corresponding to 8254 patients, between October 2010 and March 2015. A total of 5593 (66.9%) patients received an increasing E dose (ID) (oral: 2 mg/day day(d)1-7, 4 mg days d8-12, 6 mg d13-embryo transfer; transdermal: 75 µg/3 days on d1-6, 150 µg/3 days d7-embryo transfer) while 2769 (33.1%) received a constant dose (CD) of estrogen (oral: 6 mg/day 1-embryo transfer; transdermal: 150 µg/3 days d1-embryo transfer). Embryos were generated by ICSI with fresh or vitrified donor oocytes fertilized with either fresh or frozen sperm from either the couple partner or donor.PARTICIPANTS/MATERIALS, SETTING, METHODS:Cohort allocation was not related to patient characteristics; instead it reflected an internal policy change in E administration. Effect of estrogen dose (ID versus CD) on biochemical, clinical, ongoing and live birth rates, stratified by administration route, was analyzed by univariate and multivariate analysis adjusted by donor and recipient demographic and cycle characteristics.MAIN RESULTS AND THE ROLE OF CHANCE:No difference in live birth rate was found between CD and ID for oral (33.0 versus 32.5%, P = 0.81) and transdermal (35.3 versus 33.5%, P = 0.33) supplementation. Biochemical pregnancy rate was higher in CD than ID (53.7 versus 47.5%, P < 0.001) when patients received oral supplementation. Adjusted analysis confirmed that oral administration had a greater impact on biochemical pregnancy rates than transdermal (odds ratio (OR) 1.28; 95% confidence interval (CI) 1.11-1.48, P = 0.001 versus OR 1.13; 95% CI 1.00-1.30, P = 0.055). Sub-analysis of transfers between day 12 and 15 of estrogen supplementation showed no difference between CD and ID in pregnancy outcomes. Demographic variables and cycle characteristics were comparable between both groups. Moreover, the use of the oocyte donation model reduces confounding factors related to oocyte age, embryo aneuploidy, and embryo quality.LIMITATIONS, REASONS FOR CAUTION:The greatest limitation of this study is its retrospective nature. On the other hand, this study was performed using donated oocytes; although this is unlikely to affect the results, we cannot exclude the possibility that a high quality female gamete responds differently to endometrial state in comparison to a patient's own oocytes.WIDER IMPLICATIONS OF THE FINDINGS:In fresh embryo transfer from oocyte donation cycles, changes in the protocol of E replacement do not seem to have an impact on clinical outcomes and performance; for this reason estrogen replacement protocols can be adjusted to the patient's characteristics and preferences as well as to the most cost effective strategy.STUDY FUNDING/COMPETING INTERESTS:None.

Depuis 1984, lorsque le premier bebe issu d’un don d’ovocytes (DO) est ne, le nombre de femmes tombees enceintes grâce a ce traitement a constamment augmente, partout dans le monde. De bons resultats reproductifs ont rapidement ete obtenus mais, au fil du temps, certains risques ont ete identifies, principalement lies au deroulement de la grossesse. Dans cette revue, nous discutons, entre autres, de l’incidence de l’hypertension (PIH), de la pre-eclampsie (PE), de la cesarienne et des complications neonatales dans les grossesses avec DO. La comprehension de la physiopathologie des grossesses apres DO, l’identification plus precise de la sous-population des receveuses d’ovocytes presentant un risque de complications, en diminuant les grossesses multiples et en mettant en place un suivi frequent pendant la grossesse, sont autant de mesures qui reduiront les risques et permettront d’obtenir des grossesses et des naissances saines.

STUDY QUESTION:Does switching to donor semen after at least three failed oocyte donation (OD) cycles with the partner normozoospermic semen increase the live birth rate in a subsequent OD cycle? SUMMARY ANSWER:Switching to donor semen after at least three failed OD cycles with the partner normozoospermic semen does not increase the live birth rate. WHAT IS ALREADY KNOWN:In some patients, a viable pregnancy cannot be achieved after several OD cycles, despite normal diagnostic findings for the couple. The ESHRE Capri Workshop Group indicates that, in order to improve reproductive outcomes, a semen donation can be offered after three failed ICSI cycles. STUDY DESIGN, SIZE, DURATION:A retrospective cohort analysis of fourth and fifth OD cycles with either the partner's normozoospermic semen (OD) or double-donation cycles (DD), performed between January 2011 and December 2014 in a private fertility center. These couples did not have a known male factor. PARTICIPANTS/MATERIALS, SETTING, METHOD:The study included 228 cycles (159 OD and 69 DD). The fertilization method was ICSI in all cycles and embryos were transferred fresh. Fertilization rates were compared between groups using ANOVA while pregnancy outcomes were compared using Chi-square tests. Effect of DD on pregnancy outcomes was further analyzed using a logistic regression model adjusted for recipient's age and BMI, number of embryos transferred, day of embryo transfer and morphological embryo quality score. MAIN RESULTS AND THE ROLE OF CHANCE:There was no difference in live birth rate between the DD and OD groups (38.2 versus 35.8%, P = 0.73), even after adjustment for confounding factors (odds ratio 1.41, 95% confidence interval 0.72, 2.76; P = 0.31). Rates of biochemical pregnancy (52.2 versus 54.1%, P = 0.79), clinical pregnancy (41.2 versus 45.9%, P = 0.51) and ongoing pregnancy (38.2 versus 37.1%, P = 0.87) were not different between the DD and the OD groups, as well as fertilization rate (75.3 versus 75.2%, P = 0.97). The DD and OD groups were comparable at baseline in all demographic and cycle variables analyzed (recipient's BMI, number of transferred embryos and embryo quality) with the exception of recipient's age (42.3 in DD versus 44.1 in OD, P = 0.005), and day of embryo transfer (56.5% of DD and 83.6% of OD embryo transfers were performed on blastocyst stage, P < 0.001); both variables were adjusted for in the multivariate analysis. LIMITATIONS, REASONS FOR CAUTION:The main limitations of this study are its retrospective nature, the relatively small sample size, the transfer of embryos of different developmental stages and the lack of extensive molecular testing, such as sperm DNA fragmentation test, in normozoospermic patients. WIDER IMPLICATIONS OF THE FINDINGS:After excluding several causes for the failed OD cycles, the partner's normozoospermic semen was a common factor in all of them. Nevertheless, the change to a donor's semen does not seem to improve the reproductive outcomes in the subsequent cycle. STUDY FUNDING/COMPETING INTERESTS:No extra-mural funding was obtained for this study. There are no conflicts of interest to declare. TRIAL REGISTRATION NUMBER:NA.

ObjectiveThe primary objective of this study was to evaluate if body mass index (BMI) in donors, recipients and male partners was associated with live birth in oocyte donation cycles. The secondary objectives were to evaluate the association of donors, recipients and male partner BMI at the time of treatment with ovarian response to stimulation, laboratory outcome and early pregnancy events.DesignThis retrospective cohort study encompassed all oocyte donation cycles with partner sperm performed between 2010 and 2014 in a large fertility center. A total of 3,323 donors and 9,238 couples undergoing 11,806 oocyte reception cycles were included.Materials and MethodsBMI was categorized for each party as follows: underweight (BMI<18.5 kg/m2), normoweight (18.5-24.9 kg/m2), overweight (25-29.9 kg/m2), obese (30-34.9 kg/m2), and severely obese (≥35 kg/m2). The main outcome of the study was live birth, which was analyzed by univariate and multivariate analysis, adjusted for recipients' race, status of sperm, day of ET, number of transferred embryos and embryo quality. Secondary outcomes were ovarian response to stimulation, biochemical, clinical, and ongoing pregnancy.ResultsMean age was 26.1 years (4.8), 41.7 (4.7), 42.4 (6.9) in donors, recipients and male partners respectively. Mean (SD) BMI was 22.7 kg/m2 (3.2), 23.7 (4.3), 25.6 (3.5) in donors, recipients and male partners respectively. Live birth rate was 37.9%, 33.7%, 32.3%, 23.9% and 21.7% for underweight, normoweight, overweight, and obese recipients, respectively. Live birth rate in recipients was slightly but significantly affected by BMI in donors (OR 0.98 [0.96-0.99]), recipients (OR 0.98 [0.97-0.99]) and male partners (OR 0.98 [0.97-1]). Biochemical, clinical, and ongoing pregnancy rates were similarly slightly affected by BMI. We obtained 14.8 (SD 7.3), 15.0 (SD 7.5), 14.3 (SD 6.9), 12.7 mature oocytes (SD 6.1) for underweight, normoweight and overweight donors, respectively. Ovarian response to stimulation was significantly reduced in underweight compared to normoweight donors after adjustment (B=-1.52. 96%CI -2.83, -0.21; p<0.001), but not in overweight or obese ones.ConclusionsWe found a weak but statistically significant negative association between BMI and live birth rate in oocyte donation cycles. However, the clinical relevance of this association remains to be defined. ObjectiveThe primary objective of this study was to evaluate if body mass index (BMI) in donors, recipients and male partners was associated with live birth in oocyte donation cycles. The secondary objectives were to evaluate the association of donors, recipients and male partner BMI at the time of treatment with ovarian response to stimulation, laboratory outcome and early pregnancy events. The primary objective of this study was to evaluate if body mass index (BMI) in donors, recipients and male partners was associated with live birth in oocyte donation cycles. The secondary objectives were to evaluate the association of donors, recipients and male partner BMI at the time of treatment with ovarian response to stimulation, laboratory outcome and early pregnancy events. DesignThis retrospective cohort study encompassed all oocyte donation cycles with partner sperm performed between 2010 and 2014 in a large fertility center. A total of 3,323 donors and 9,238 couples undergoing 11,806 oocyte reception cycles were included. This retrospective cohort study encompassed all oocyte donation cycles with partner sperm performed between 2010 and 2014 in a large fertility center. A total of 3,323 donors and 9,238 couples undergoing 11,806 oocyte reception cycles were included. Materials and MethodsBMI was categorized for each party as follows: underweight (BMI<18.5 kg/m2), normoweight (18.5-24.9 kg/m2), overweight (25-29.9 kg/m2), obese (30-34.9 kg/m2), and severely obese (≥35 kg/m2). The main outcome of the study was live birth, which was analyzed by univariate and multivariate analysis, adjusted for recipients' race, status of sperm, day of ET, number of transferred embryos and embryo quality. Secondary outcomes were ovarian response to stimulation, biochemical, clinical, and ongoing pregnancy. BMI was categorized for each party as follows: underweight (BMI<18.5 kg/m2), normoweight (18.5-24.9 kg/m2), overweight (25-29.9 kg/m2), obese (30-34.9 kg/m2), and severely obese (≥35 kg/m2). The main outcome of the study was live birth, which was analyzed by univariate and multivariate analysis, adjusted for recipients' race, status of sperm, day of ET, number of transferred embryos and embryo quality. Secondary outcomes were ovarian response to stimulation, biochemical, clinical, and ongoing pregnancy. ResultsMean age was 26.1 years (4.8), 41.7 (4.7), 42.4 (6.9) in donors, recipients and male partners respectively. Mean (SD) BMI was 22.7 kg/m2 (3.2), 23.7 (4.3), 25.6 (3.5) in donors, recipients and male partners respectively. Live birth rate was 37.9%, 33.7%, 32.3%, 23.9% and 21.7% for underweight, normoweight, overweight, and obese recipients, respectively. Live birth rate in recipients was slightly but significantly affected by BMI in donors (OR 0.98 [0.96-0.99]), recipients (OR 0.98 [0.97-0.99]) and male partners (OR 0.98 [0.97-1]). Biochemical, clinical, and ongoing pregnancy rates were similarly slightly affected by BMI. We obtained 14.8 (SD 7.3), 15.0 (SD 7.5), 14.3 (SD 6.9), 12.7 mature oocytes (SD 6.1) for underweight, normoweight and overweight donors, respectively. Ovarian response to stimulation was significantly reduced in underweight compared to normoweight donors after adjustment (B=-1.52. 96%CI -2.83, -0.21; p<0.001), but not in overweight or obese ones. Mean age was 26.1 years (4.8), 41.7 (4.7), 42.4 (6.9) in donors, recipients and male partners respectively. Mean (SD) BMI was 22.7 kg/m2 (3.2), 23.7 (4.3), 25.6 (3.5) in donors, recipients and male partners respectively. Live birth rate was 37.9%, 33.7%, 32.3%, 23.9% and 21.7% for underweight, normoweight, overweight, and obese recipients, respectively. Live birth rate in recipients was slightly but significantly affected by BMI in donors (OR 0.98 [0.96-0.99]), recipients (OR 0.98 [0.97-0.99]) and male partners (OR 0.98 [0.97-1]). Biochemical, clinical, and ongoing pregnancy rates were similarly slightly affected by BMI. We obtained 14.8 (SD 7.3), 15.0 (SD 7.5), 14.3 (SD 6.9), 12.7 mature oocytes (SD 6.1) for underweight, normoweight and overweight donors, respectively. Ovarian response to stimulation was significantly reduced in underweight compared to normoweight donors after adjustment (B=-1.52. 96%CI -2.83, -0.21; p<0.001), but not in overweight or obese ones. ConclusionsWe found a weak but statistically significant negative association between BMI and live birth rate in oocyte donation cycles. However, the clinical relevance of this association remains to be defined. We found a weak but statistically significant negative association between BMI and live birth rate in oocyte donation cycles. However, the clinical relevance of this association remains to be defined.

The availability of preconceptional screening for X-linked and recessive diseases offers the opportunity to lower the risk to conceive an affected child. Genetic matching of donors and patients further improves ART service; however, the donor selection process might alter expected frequencies of certain diseases, potentially leading to variations of pretest risk. Fragile X syndrome (FXS) is the most common cause of inherited intellectual disability affecting approximately 1:4000-1:6000 births. FXS is caused by the expansion of CGG repeats in the FMR1 gene, and depending on CGG repeats number the population has been classified into: normal (N) 5-44 repeats, intermediate allele (IA) 45-54, premutation (PM) 55-200, and full mutation (FM) >201. People with PM or FM could develop clinical symptoms and transmit the disease to the next generation. IA alleles do not confer an increased FXS risk for the very next generation, however, the 50-54 range may show instability, with potential expansions in subsequent generations. The aim of the study is to assess carrier status and ethnic variation of the FMR1 gene in oocyte donation (OD) candidates. Retrospective cohort study. 336 consecutive OD candidates aged 18 to 35 years were tested for FXS (Asuragen Amplidex FMR1) in January-April 2015. Women with a family history of FXS, mental retardation, chromosomal abnormalities, genetics disorders or neurological conditions were excluded from further screening. All women diagnosed with 45 o more CGG repeats were referred to genetic counseling. Relative frequency of IA status by ethnicity was assessed by Chi2 test, while the association between IA status and ovarian reserve was assessed using a logistic regression model, adjusted by age. Women ethnicities were: Mediterranean 127 (37.8%), European 103 (30.6%), Latino American 61 (18.2%), Caribbean 27 (8.0%), other ethnicities 18 (5.4%). There were 17 carriers (1:20 overall), all of them IA; 13 carriers had 45-49 CGG, while 4 had 50-54 CGG. The frequency of IA was 7.4%, 7.8%, and 5.5% in women of Caribbean, European, and Mediterranean ethnicity, respectively (p>.05). IA status was not associated with the woman ovarian reserve (p>.05). Screening of potential donors for family history of FXS-like phenotypes lowers the frequency of PM and FM alleles in accepted donors. The high frequency of IA alleles in Caribbean, Europeans, and Mediterranean ethnicity is likely due to ethnic and geographical variability, a phenomenon described in other subgroup population.

DNA damage in cumulus cells (CCs) might be related with the developmental competence of the enclosed oocytes, however, conclusive studies are missing, partially due to the lack of a reliable, cheap, fast, and reproducible DNA damage test. We report the development of a chromatin dispersion test that allows for a fast evaluation of double strand DNA (ds-DNA) damage in CCs. The whole experiment was performed using CCs from 103 oocyte retrieval cycles evaluating the prototype D3-MAX ability (a chromatin dispersion based assay) to detect DNA breaks against in situ nick translation (ISNT) and a two tailed comet assay (TT-comet). Samples were collected from women younger than 35 years of age with a good response to stimulation. Pooled cumulus cells of MII oocytes were used. The chromatin dispersion assay results correlate with the double strand-DNA breaks values assessed by the TT-comet assay (Spearman Rho = 0.624; p = 0.003;), while the correlation was poor when compared to the single strand DNA (ss-DNA) breaks observed also with the TT-comet assay (Spearman Rho = -0.141; p = 0.554). ISNT showed a correspondence in the same cells between enzymatic incorporation of modified nucleotides and halos of chromatin dispersion. We conclude that D3-Max test detects mainly ds-DNA breaks in cumulus cells and is a reliable, fast, and easy reproducible assay suitable for routine clinical practices once the influence on oocyte quality has been established.