Semi-automated versus manual embryo vitrification: inter-operator variability, time-saving, and clinical outcomes.

PURPOSE: Does semi-automated vitrification have lower inter-operator variability and better clinical outcomes than manual vitrification? METHODS: Retrospective analyses of 282 patients whose embryos had been cryopreserved, manually with Irvine®-CBS® (MV) or semi-automatically vitrified with the GAVI® method (AV) (from November 2017 to September 2020). Both techniques were performed during the same period by 5 operators. Inter-operator variability was statistically analyzed between operators who performed the vitrification and those who performed the warming process to compare the intact survival rate (% embryos with 100% intact blastomeres) and the positive survival rate (at least 50% intact blastomeres). Additionally, the complete vitrification time was assessed for the 2 techniques according to the number of vitrified embryos. RESULTS: Manual vitrification involved warming 338 embryos in 266 cycles for 181 couples compared to 212 embryos in 162 AV cycles for 101 patients. The positive survival rate was higher (p < 0.05) after MV (96%; 323/338) than after AV (90%; 191/212). The intact survival rate (86 vs 84%) and the clinical pregnancy rate (27 vs 22%) were not significantly different between MV and AV. Regarding the inter-operator variability, no significant difference in positive and intact survival rate was evident between the 5 technicians, neither by vitrification nor by warming steps with MV and AV. Concerning time-saving, the MV technique proved to be quicker than AV (minus 11 ± 9 min). CONCLUSIONS: Manual vitrification exhibited favorable total survival rates and was more time efficient, while both MV and AV cooling and warming treatments showed little operator variability.

Spontaneous pregnancy rate following surgery for deep infiltrating endometriosis in infertile women: The impact of the learning curve.

OBJECTIVES: To determine whether the surgical learning curve impact the spontaneous pregnancy rate in infertile patients undergoing removal of deep infiltrating endometriosis MATERIAL AND METHODS: Single center retrospective study including the first 50 consecutive infertile women suffering from deep infiltrating endometriosis and referred to a single surgeon. All patients underwent laparoscopic removal of deep endometriosis lesions. The study population was stratified in two subgroups, namely the early group (including the first 25 cases) and the late group (comprising the 25 subsequent cases). Pregnancy and live birth rates, surgical morbidity and clinical recurrence rate were compared between study groups. RESULTS: Overall, spontaneous pregnancy rate (40 % in the early group versus 56 % in the late group, p = 0.25), live birth rate (40 % versus 44 %, p = 0.77) and clinical recurrence rate (16 % versus 4%, p = 0.16) did not significantly differ between the study groups. Logistic regression analysis revealed that ASRM stage, EFI score, and body mass index were the only significant prognostic factors of postoperative spontaneous fertility. CONCLUSION: Surgical resection of deep infiltrating endometriosis in infertile women is associated with high spontaneous pregnancy and live birth rates. The surgeon's learning curve does not impact postoperative fertility outcomes.

Fertility and sexuality of women with inflammatory arthritis.

OBJECTIVE: This study aimed to evaluate the impact of the four main types of inflammatory arthritis (rheumatoid arthritis (RA), ankylosing spondylitis (AS), psoriatic arthritis (PA), juvenile chronic arthritis (JCA)) on female fertility. DESIGN: We performed a monocentric observational study, which took place in the Reproductive Medicine Department and the Rheumatology Department of the Toulouse University Hospital. An anonymous questionnaire was sent to patients aged 18-50 with RA, AS, PA or JCA who were diagnosed before the age of 40 and who consented to participate in the study. A section of the questionnaire was completed using the medical file of each patient. RESULTS: Of the 521 eligible patients, 202 (39 %) answered the questionnaire. Before being diagnosed with a rheumatic disease, 87 % of patients with a desire for pregnancy had a spontaneously obtained live birth, compared to 75 % after (P < 0.05). However, this decrease only concerned RA (90 % before diagnosis vs 68 % after; P < 0.05). This difference was due to a lower rate of pregnancies achieved, but the number of miscarriages did not increase. Moreover, for patients who managed to conceive, the time required for conception did not increase, regardless of the type of rheumatic disease they had. Two thirds of patients reported a decrease in the frequency of sexual intercourse after diagnosis, mainly (75 %) because of the pain relating to the disease. CONCLUSION: Our study shows a trend of decreased fertility in patients with RA, perhaps due to sexual problems, while no impact of AS and PA on the ability to conceive was found. This impact on fertility didn't seem to be related to the decrease of sexuality. Multidisciplinary care including treatment from a rheumatologist, gynecologist and sexologist would appear essential in order to provide RA patients with the necessary information concerning their fertility, answer their questions about pregnancy and thus optimize their care.

Need for choosing the ideal pH value for IVF culture media.

PURPOSE: Monitoring the pH of IVF culture media is a good practice, but the required pH levels have been "arbitrarily" set. Assisted reproductive technology centers around the world are spending time and money on pH monitoring without any consensus to date. The objective of this narrative review was to evaluate the importance of pH monitoring during IVF, discover how the oocyte and embryo regulate their intracellular pH and try to determine the optimal pH to be applied. METHODS: A narrative literature review was performed on publications in the PubMed database reporting on the impact of pH on cellular function, oocyte and embryo development, IVF outcomes and pathophysiology, or on physiological pH in the female reproductive tract. RESULTS: Intracellular pH regulates many cellular processes such as meiotic spindle stability of the oocyte, cell division and differentiation, embryo enzymatic activities, and blastocoel formation. The internal pH of the human embryo is maintained by regulatory mechanisms (mainly Na+/H+ and HCO3-/Cl- exchangers) that can be exceeded, particularly in the oocyte and early-stage embryos. The opinion that the optimal pH for embryo culture is physiological pH is not correct since several physicochemical parameters specific to IVF culture conditions (temperature, medium composition, duration of culture, or implication of CO2) can modify the intracellular pH of the embryo and change its needs and adaptability. CONCLUSIONS: Because correct and stable extracellular pH is essential to embryo health and development, monitoring pH is imperative. However, there is a lack of clinical data on choosing the ideal pH for human IVF culture media.

Potential chances for natural fertility influence results of intrauterine inseminations.

OBJECTIVE: Intra-uterine insemination (IUI) is widely used for different indications. The aim of the present study was to evaluate the efficiency of intra-uterine insemination as a function of indication and origin of the inseminated spermatozoa. STUDY DESIGN: The retrospective study involved 827 first attempts of IUI in 827 couples between January 2011 and July 2017 in the Toulouse university hospital. Of these, 642 used fresh sperm from the husband, 40 frozen sperm from the husband and 145 frozen sperm from donors. The measured outcome parameter was live birth rate per attempt. RESULTS: When comparing couples lacking functional gametes (due to male or female causes), to couples who could potentially conceive spontaneously, i.e. subfertile, the latter were found to have a significantly lower live birth rate (18% vs 26%; P < 0.05). Even when adjusted for demographic parameters, which differed significantly between the 2 groups (female age, percentage of women suffering from primary infertility, BMI, number of inseminated motile spermatozoa and stimulation duration), this difference remained statistically significant (OR = 0.639 [0.425-0.961]; P = 0.0316). CONCLUSION: When compared to couples lacking functional gametes, subfertile couples have poor IUI outcomes, suggesting a hidden cause of infertility, despite no apparent differences in ovarian reserve, tubal potency, results of ovarian stimulation and normal conventional sperm parameters. Further studies are required to better characterise and identify this subgroup of women with poor IUI outcomes.

Age-specific anti-Mullerian hormone (AMH) levels poorly affects cumulative live birth rate after intra-uterine insemination.

OBJECTIVE: To evaluate the impact of age-specific anti-Mullerian (AMH) levels on the cumulative live birth rate after 4 intra uterine inseminations (IUI). STUDY DESIGN: The retrospective study study involved 509 couples who underwent their first IUI between January 2011 and July 2017 in the Toulouse University Hospital. All IUI were performed after an ovarian stimulation combining recombinant FSH and GnRH antagonist. The main measure outcome was the cumulative live birth rate (LBR) defined as the number of deliveries with at least one live birth resulting from a maximum of 4 IUI attempts. RESULTS: When compared to normal or high levels, low age-specific AMH (<25th of the AMH in each age group) was associated to a non-significant lower live birth rate (31%, 38% and 42% respectively for low, normal and high age-specific groups; P = 0.170) and non-significant higher miscarriage rate (26%; 19% and 14% respectively for low, normal and high age-specific groups; P = 0.209). However, it must be pointed out that in low age-specific AMH the initial FSH doses used for stimulation were higher than in the other groups. CONCLUSION: This study shows that the age-specific levels of AMH have only a slight effect on IUI outcome when adapting the stimulation protocols to their level.

Fertility of women with cystic fibrosis: a French survey.

RESEARCH QUESTION: Although the impact of cystic fibrosis on male fertility is well known, very few studies have investigated its effect on female fertility. This study aimed to evaluate the fertility status of women with cystic fibrosis. DESIGN: A questionnaire was sent to 220 women with cystic fibrosis. The questions concerned their desire to become a parent, achievement or not of a pregnancy, the time to become pregnant, the means of achieving pregnancy (spontaneously or with medical assistance) and the outcome of the pregnancy. Ninety-eight patients responded to the questionnaire. RESULTS: Of the 46 women who sought pregnancy, 25 (54%) had at least one live birth without treatment, while 11 (24%) required infertility treatment to obtain a live birth and 10 (22%) had no delivery. The mean time-to-pregnancy was 12 months (1-180). The reasons for preferring not to become pregnant were mainly fear of the interaction between cystic fibrosis and pregnancy and of the transmission of cystic fibrosis to children. CONCLUSIONS: Fertility seems to be slightly impaired in women with cystic fibrosis, because 37% of them failed to become pregnant without medical assistance. Because the outcome of pregnancies appears normal, patients should be informed about the possibility of becoming mothers and be made aware of the risk of unwanted pregnancies.

Association between progesterone to number of mature oocytes index and live birth in GnRH antagonist protocols.

RESEARCH QUESTION: What is the association between blood progesterone level alone, progesterone to mature oocytes index (PMOI) and IVF outcomes? DESIGN: Clinical data from 960 couples undergoing their first fresh embryo transfer after an intracytoplasmic sperm injection attempt carried out between September 2012 and July 2017 were analysed. All patients underwent ovarian stimulation combining recombinant FSH and gonadotrophin releasing hormone antagonist. Progesterone was measured on the day on which ovulation was triggered. The PMOI was divided into four groups based on 25th, 50th and 75th percentiles, and live birth and implantation rates were compared between the groups. RESULTS: A negative association was found between PMOI levels, live birth and implantation rates. When adjusting for age, ovarian stimulation index (OSI) and number of embryos transferred, the PMOI remained negatively correlated to live birth rate (LBR) (OR = 0.147 [0.031 to 0.701]; P = 0.0161), whereas total blood progesterone was no longer evident. Moreover, the LBR (18.5% versus 28.4%; P < 0.01) and implantation rate (12.4% versus 21.0%; P < 0.01) were significantly decreased only when PMOI was 0.167 ng/ml or greater, irrespective of progesterone concentration (< or ≥1.08 ng/ml). The opposite was not true, however. Similar PMOIs were recorded in the same patient from one attempt to the next, and were partially linked to basal FSH, anti-Müllerian hormone, antral follicle count and OSI. CONCLUSIONS: PMOI seems to be more predictive than total progesterone level of IVF outcome and reflects the ability of embryos to develop as it is linked to the implantation rate.

Impact of estradiol and progesterone levels during the late follicular stage on the outcome of GnRH antagonist protocols.

The study aimed to assess the impacts and the targets of progesterone (P4) and estradiol (E2) levels on IVF outcomes in GnRH antagonist protocols. The study was retrospective and concerned patients for their first fresh embryo transfers, after stimulation by a recombinant FSH (rFSH)-GnRH antagonist protocol, between September 2012 and July 2017 in the Toulouse University Hospital. Multivariable analysis, taking into account female age and the ovarian stimulation index, showed that E2 levels had no impact on IVF outcomes, while high P4 levels (>1.10 ng/mL) were associated to low pregnancy rate. The P4 concentrations were significantly negatively correlated to the percentage of top embryos and to the implantation rate. Therefore, the deleterious effect of high levels P4 could to act mainly by impairing embryo quality, which questions the place of the freeze-all strategy in these cases.

Decrease in pregnancy rate after endometrial scratch in women undergoing a first or second in vitro fertilization. A multicenter randomized controlled trial.

STUDY QUESTION: Does endometrial scratch in women undergoing a first or second IVF/ICSI attempt improve the clinical pregnancy rate (CPR)? SUMMARY ANSWER: Endometrial scratch (ES) in women undergoing their first or second IVF/ICSI attempt does not enhance the CPR under the technical conditions of our study. WHAT IS KNOWN ALREADY: Several studies have suggested that physical scratch of the endometrium before an IVF attempt could improve embryo implantation. STUDY DESIGN, SIZE, DURATION: This was a randomized controlled multi-center, two-arm, parallel trial. Inclusions started in February 2010 and stopped prematurely in July 2014 after an unplanned interim analysis. At the time of study closure, 191 of the planned 358 patients had been included. PARTICIPANTS/MATERIALS, SETTING, METHODS: Patients included in the study were randomly assigned to either the ES arm or the non-ES arm. Local ES was performed between Day 20 and Day 24 of the cycle preceding ovarian stimulation using a device for endometrial biopsy. Ovarian stimulation used a combination of recombinant FSH and either an GnRH agonist protocol or a GnRH antagonist protocol without any estrogen pre-treatment. CPR was analyzed on an intent-to-treat basis. All comparisons between the two groups were done using a logistic regression model adjusted for age, BMI and infertility etiology. Differences between the two arms were considered statistically significant at P value of less than 0.0446 for the primary outcome only. MAIN RESULTS AND THE ROLE OF CHANCE: Sixty-eight embryo transfers were performed in the ES arm and sixty-four in the non-ES arm. CPR was 23.5% (16/68) in the ES arm and 35.9% (23/64) in the non-ES arm (hazard ratio (HR) = 0.43; 95% CI, 0.18-1.02; P = 0.0568). The implantation rate was 19.1% and 24.0% in the ES arm and in the non-ES arm, respectively. Two miscarriages and one ectopic pregnancy were reported in each arm. The multiple pregnancy rate was higher in the scratch arm (50.0% vs 20.0%), but the difference was not statistically significant (odds ratio (OR) = 4.54; 95% CI, 0.50-40.93; P = 0.1349). The endometrial biopsy procedure was well tolerated in most women. Of 50 patients in the ES arm having received the embryo transfer, 40 (80.0%) patients reported having felt pain during the procedure, the pain resolving quickly for 31 of them. LIMITATIONS, REASONS FOR CAUTION: An interim analysis of the primary endpoint was conducted and an independent data monitoring committee agreed on stopping the inclusions. This analysis was prompted by the tendency towards lower pregnancy rates observed in the ES arm. Consequently, the study suffered from a lower inclusion rate and failed to reach the planned sample size. WIDER IMPLICATIONS OF THE FINDINGS: Under the technical condition employed in this study, ES in the luteal phase of the cycle preceding the ovarian stimulation does not improve CPR in patients undergoing a first or second IVF/ICSI attempt. STUDY FUNDING/COMPETING INTERESTS: This study was supported by a grant from Ministère de la Santé Français (Programme Hospitalier de Recherche Clinique 2009). There are no conflicts of interest. TRIAL REGISTRATION NUMBER: NCT01064193. TRIAL REGISTRATION DATE: 08-Feb-2010. DATE OF FIRST PATIENT'S ENROLMENT: 08-Feb-2010.

Anti-sperm antibodies detection by a modified MAR test: Towards a better definition of its indications.

RESEARCH QUESTION: Anti-sperm antibodies (ASA) have been shown to reduce male fertility but consensus about the precise situations in which tests should be carried out are lacking. In infertility investigations, should the mixed antiglobulin reaction (MAR) test be a first-line test? Should it be carried out systematically before assisted reproductive technology (ART)? What are the risk factors for ASA? DESIGN: All infertile patients (n = 1364) were tested with SpermMar (modified MAR test) between July 2013 and June 2017. Intra-patient variability of the MAR test was also assesed by comparing two tests within the same year in selected patients (n = 101). RESULTS: The main factor that influenced the percentage of ASA was the presence or absence of sperm agglutination. In the presence of agglutinations, 27 out of 72 (37.5%) patients were positive for ASA compared with 33 out of 1292 (2.6%) in the absence of agglutinations (P < 0.0001). When one risk factor was present (spontaneous sperm agglutination, history of scrotal trauma or inguinal surgery), 33 out of 179 (18.44%) tests were positive for ASA (≥50% coated spermatozoa), whereas only 27 out of 1242 (2.2%) were positive when no risk factor was present (P < 0.0001). CONCLUSIONS: ASA detection should not be systematically recommended in investigations of fertility status and before ART but reserved for when sperm agglutination is found during conventional sperm examination, or if the patient has a history of scrotal trauma or has undergone inguinal surgery.

Pregnancy outcome in Turner syndrome: A French multi-center study after the 2009 guidelines.

OBJECTIVE: This study aimed to assess the application of the French guidelines for pregnancies in Turner syndrome (TS) and their impact on perinatal prognosis. STUDY DESIGN: We performed a French multi-center retrospective study (14 centers), including TS pregnant patients (spontaneously or by Assisted Reproductive Technology (ART)) between January 2006 and July 2017. Only clinical pregnancies were analyzed. The adjustment of medical follow-up modalities to French guidelines was evaluated for all pregnancies after 2009. Pregnancies from oocyte donation (OD) after 2009 were compared to those of a cohort of TS pregnancies obtained by OD before 2009, which were reported by the French Study Group for Oocyte Donation. RESULTS: One hundred seventy pregnancies in 103 patients were included: 35 spontaneous, 5 by means of intra-conjugal ART, and 130 with OD. No serious maternal complications were observed. We reported two stillbirths and one intra uterine fetal death. The French guidelines were partially respected. The preconceptional assessment was carried out in 74% of cases. Cardiology follow-up during pregnancy was performed in accordance with guidelines in 74% of patients. Postpartum cardiac ultrasonography was performed in 45% of pregnancies but only in 11% within 8 days post-partum. When compared to the 2009 historical cohort, the rates of high blood pressure (19% vs. 38%; p < 0.005) pre-eclampsia (8% vs. 21%; p < 0.005) and prematurity <35 weeks (15% vs 38%; p < 0.0001) were lower. CONCLUSIONS: The implementation of guidelines has allowed the standardization of TS pregnancy care and improved perinatal indicators for both mothers and children. However, an effort must be done, in a postpartum survey.

Mother's age at menopause but not own age at menarche has an impact on ovarian reserve.

To detect clinical parameters impacting ovarian reserve, data were analyzed from 573 patients who had an anti-Müllerian hormone (AMH) measurement for infertility treatment. No impact was found on the age at menarche but a significant diminished ovarian reserve was observed when a patient's mother was menopausal before age 50. These data suggest that ovarian reserve must be monitored in such patients to offer them fertility preservation when at risk of premature ovarian insufficiency (POI).

Establishment and validation of a score to predict ovarian response to stimulation in IVF.

This study aimed to integrate clinical and biological parameters in a score able to predict ovarian response to stimulation for IVF in gonadotrophin-releasing hormone (GnRH) antagonist protocols. A progressive discriminant analysis to establish a score including the main clinical and biological parameters predicting ovarian response was performed by retrospectively analysing data from the first ovarian stimulation cycle of 494 patients. The score was validated in a prospectively enrolled, independent set of 257 patients undergoing their first ovarian stimulation cycle. All ovarian stimulations were performed using a combination of GnRH antagonist and recombinant FSH. Ovarian response was assessed through ovarian sensitivity index (OSI). Parameters from the patients' database were classified according to correlation with OSI: the progressive discriminant analysis resulted in the following calculation: score = 0.192 - (0.004 × FSH (IU/l)) + (0.012 × LH:FSH ratio) + (0.002 × AMH (ng/ml)) - (0.002 × BMI (kg/m2)) + (0.001 × AFC) - (0.002 × age (years)). This score was significantly correlated with OSI in the retrospective (r = 0.599; P < 0.0001) and prospective (r = 0.584; P < 0.0001) studies. In conclusion, the score including clinical and biological parameters could explain 60% of the variance in ovarian response to stimulation.

Does air pollution play a role in infertility?: a systematic review.

BACKGROUND: Air pollution is involved in many pathologies. These pollutants act through several mechanisms that can affect numerous physiological functions, including reproduction: as endocrine disruptors or reactive oxygen species inducers, and through the formation of DNA adducts and/or epigenetic modifications. We conducted a systematic review of the published literature on the impact of air pollution on reproductive function. Eligible studies were selected from an electronic literature search from the PUBMED database from January 2000 to February 2016 and associated references in published studies. Search terms included (1) ovary or follicle or oocyte or testis or testicular or sperm or spermatozoa or fertility or infertility and (2) air quality or O3 or NO2 or PM2.5 or diesel or SO2 or traffic or PM10 or air pollution or air pollutants. The literature search was conducted in accordance with the Preferred Reporting Items for Systematic Reviews and Meta-Analyses (PRISMA) guidelines. We have included the human and animal studies corresponding to the search terms and published in English. We have excluded articles whose results did not concern fertility or gamete function and those focused on cancer or allergy. We have also excluded genetic, auto-immune or iatrogenic causes of reduced reproduction function from our analysis. Finally, we have excluded animal data that does not concern mammals and studies based on results from in vitro culture. Data have been grouped according to the studied pollutants in order to synthetize their impact on fertility and the molecular pathways involved. CONCLUSION: Both animal and human epidemiological studies support the idea that air pollutants cause defects during gametogenesis leading to a drop in reproductive capacities in exposed populations. Air quality has an impact on overall health as well as on the reproductive function, so increased awareness of environmental protection issues is needed among the general public and the authorities.

Effect of unilateral tubal abnormalities on the results of intrauterine inseminations.

A total of 101 patients with one normal tube were compared with 117 patients with two normal tubes to assess the effect of unilateral tubal abnormalities on the results of intrauterine inseminations. The clinical pregnancy and live birth rates seemed to reduce by one-half in almost all types of abnormality, suggesting that these patients should be preferentially treated with IVF.

Ovarian Mucinous Cystadenoma After Ovarian Graft.

BACKGROUND: Freezing strips of ovarian cortex before chemotherapy followed by transplantation is an experimental method to preserve fertility for reproductive-aged women with cancer. We report a case of a cancer patient who developed a mucinous cystadenoma in a grafted piece of ovarian cortex. CASE: A 32-year-old woman with a Ewing sarcoma had ovarian cryopreservation using cortical strip freezing before receiving chemotherapy. Five years later she had no ovarian function, and the strips were thawed and grafted back onto the ovary. She spontaneously became pregnant 1 year after this procedure and delivered a healthy neonate near term. During the cesarean delivery, a 5×3-cm cyst was removed from the graft. On pathologic evaluation, it was determined to be a mucinous cystadenoma. CONCLUSION: Ovarian pathology can develop in previously frozen ovarian cortex tissue after transplantation back onto the ovary. This suggests that routine gynecologic surveillance remains important for these women.

Follicular growth but absence of oocyte and cumulus maturation during ovarian stimulation in the days following surgical abortion: a case report.

This paper is a case report on the results of an ovarian stimulation performed in the days following an induced abortion. A patient had breast cancer diagnosed during an early pregnancy. She had an induced abortion and had, before chemotherapy an ovarian stimulation, using rFSH and GnRH antagonist, followed by follicular puncture for oocyte for vitrification in the view of fertility preservation. No oocyte could be obtained despite a good hormonal and ultrasonographical follicular growth. This case report suggest that ovarian stimulation must be delayed after abortion to allow the maturation of oocyte-cumulus complexes.