Live Birth Group Research Articles

Predicting the pregnancy outcome: Uncovering the link between β-hCG, patient factors and live birth in IVF

ObjectiveThe objectives of the study were to correlate live birth rate with the initial level of β-hCG in all patients undergoing embryo transfer and develop a predictive model of live birth based on patients’ and assisted reproductive technology (ART) characteristics. Study designSingle-center retrospective cohort study, including a total of 7587 positive first serum β-hCG from women who performed fresh and frozen embryo transfer.Twenty-one variables related to patient characteristics and treatment modalities were analyzed by step-wise univariate analysis followed by multivariate analysis. The study included women between 18 and 45 years with a positive (>20 IU/L) serum β-hCG between January 2011 and December 2020 while it excluded cycles from donation of gametes, PGT and >3 embryos transferred simultaneously. Main results and the role of chance: Among the positive serum β-hCG measurements 5085 (67.0 %) resulted in live births. β-hCG was higher in the live birth group (691 IU/L) compared to the failed pregnancies group (304 IU/L) (p500 IU/mL provided a positive predictive value for live birth of 85.3 % (95 %CI 84.0–86.4).Failed pregnancies were more common among older maternal and paternal age. Previous abortions were more common in the failed pregnancies group (25.0 %) compared to the live birth group (19.4 %). The most common indication to treatment was male infertility, accounting for a larger portion in the live birth group (41.7 %). At multivariate analysis, the factors independently associated with live birth were: first serum β-hCG (OR 1.35, 95 %CI 1.32–1.37), maternal age (OR 0.93, 95 %CI 0.91–0.95), previous abortions (OR 0.77, 95 % CI 0.68–0.88). The number of embryos transferred showed a positive correlation below a maternal age of 39 (<35 years old OR 1.60, CI 95 % 1.32–1.94; <39 years old OR 1.27 CI 95 % 1.09–1.47). The prediction model incorporating these four variables resulted in a sensitivity and specificity with an area under the ROC curve of 0.798. Conclusion: The study confirms the importance of the first value of β-hCG and proposes a predictive model that takes into account maternal age, number of embryos transferred, and obstetric history, thereby aiding in couples’ counseling and ultimately improve patients’ care.

P-432 The association of early pregnancy loss and thrombophilia

Abstract Study question Is there a correlation between thrombophilia and the occurrence of pregnancy loss? Summary answer No statistically significant association is observed between thrombophilia and the live births. What is known already The understanding of recurrent pregnancy loss is a subject of controversy, encompassing both its causes and potential treatments and there is currently no consensus regarding the criteria for examining patients experiencing pregnancy loss, the specific tests to be conducted, or the appropriate treatment approaches. The connection between hereditary thrombophilia and pregnancy loss remains unclear. Study design, size, duration Within the timeframe of 01/12/2020 to 01/06/2022, pregnancy loss clinic of a tertiary university hospital received a total of 296 patients who had experienced at least one pregnancy loss. Following examination, these patients were encouraged to return to clinic if they became pregnant again. Among them, 192 patients did become pregnant and sought further care. These patients received treatment based on pregnancy loss clinic’s management algorithm, and their subsequent pregnancies were closely monitored. Participants/materials, setting, methods This prospective study involved 192 patients who, after undergoing examination, experienced subsequent pregnancies. These patients were categorized into two groups based on their obstetric outcomes: live birth and pregnancy loss. The primary focus of the study was to compare hereditary and acquired thrombophilia between these two groups. Additionally, secondary outcomes included a comparison of obstetric history, genetics, uterine anomalies, and results from biochemical and hormonal examinations between the two groups. Main results and the role of chance 150 of 192 patients achieved live birth (78.13%). The prevalence of hereditary thrombophilia mutations showed no significant differences between the two groups. Specifically, in the pregnancy loss group, the FV Leiden mutation rate was 12.5%, compared to 13.7% in the live birth group (p = 0.836). The Prothrombin gene mutation was observed in 7.5% of the pregnancy loss group and 7.25% of the live birth group (p = 1.000). Additionally, the PAI 4G/5G polymorphism was present in 60% of the pregnancy loss group and 52% of the live birth group (p = 0.347), the MTHFR/C677T polymorphism was detected in 62.5% of the pregnancy loss group and 56.5% of the live birth group (p = 0.500), and the MTHFR/1298C polymorphism was identified in 62.5% of the pregnancy loss group and 60.1% of the live birth group (p = 0.788). While antithrombin III levels were comparable between the two groups (p = 0.53), a noteworthy decline in antithrombin III levels was observed with an increasing history of losses: 110.2 for one loss, 106.0 for two losses, and 100.2 for three or more losses (p = 0.033). The homocysteine level was found to be 10.85 μM (range: 6.60-24.80 μM) in the pregnancy loss group and 9.40 μM (range: 4.90-37.00 μM) in the live birth group (p = 0.023). Limitations, reasons for caution A limitation of our study is its single-center nature. Additionally, we did not include a healthy control group with no history of pregnancy loss, preventing a direct comparison between the pregnancy loss group and a group unaffected by such experiences. Wider implications of the findings Our study provides valuable insights into questioning the live birth expectations of women who have experienced pregnancy loss and underscores the significance of thrombophilia in the context of pregnancy loss. Trial registration number not applicable

O-126 Blastocyst spent culture medium metabolite levels and reproductive outcomes

Abstract Study question Is there any relationship between blastocyst spent culture medium metabolite levels and reproductive outcomes? Summary answer Metabolomic analysis of 173 metabolites revealed differences in pregnancy, implantation, and live birth rates, highlighting specific metabolites' associations with reproductive outcomes. What is known already Identifying embryos with higher implantation potential poses a significant challenge in assisted reproductive technology (ART). Currently, morphological criteria are utilized for this selection; however, this method does not consistently offer precise predictions of successful implantation. In recent years, there are several hypothesis to employ metabolomic approaches to assess developing embryos and discriminate preferable profiles associated with embryo viability. Metabolomics involves the non-targeted identification and quantification of low molecular weight end-products of metabolism, referred to as metabolites. However, there is limited knowledge regarding the impact of metabolomic profile on implantation rates. Study design, size, duration This prospective case control study enrolled 70 patients who underwent intracytoplasmic sperm injection (ICSI) and single blastocyst transfer. Spent culture mediums (SCM) were collected in fifth day of each embryo. Metabolite levels of SCMs were measured by Liquid Chromatography-Mass Spectrometry. Metabolite levels were compared according to pregnancy, implantation and live birth rates. Participants/materials, setting, methods Normo-responder patients aged between 21 and 39, experiencing primary infertility, whose embryos progressed to the blastocyst stage and underwent single blastocyst transfer. Human Metabolome Database and Compound Discoverer 3.3 was used for the identification of the metabolites. Statistical Package for the Social Sciences (SPSS) software were used for statistical analysis. p&amp;lt;.05 was considered statistically significant. Main results and the role of chance The baseline parameters between groups were not significantly different between pregnant and non-pregnant patient groups. A total of 173 metabolites were analysed. When the metabolomics compared according to pregnancy results(68.5%), 1-(4-Methoxyphenyl)-3-pentanone, morpholine propane sulfanoic acid, naproxen were found higher in pregnant group while 4-indolecarbaldehyde, eicosapentaenoic acid, hepes were significantly higher in non-pregnant group (p&amp;lt;.05). According to the implantation (65.7%) rates, 1-(4-Methoxyphenyl)-3-pentanone was significantly higher in the implantation group. According to the live birth rates (54.2%), 1-(4-Methoxyphenyl)-3-pentanone, 15,16-Dihydroxyoctadecanoic acid, ALA-PRO, HEPES, Naproxen, pyroglutamylglycine, lactamide (p&amp;lt;.05) was higher in the live birth group. 1,2,3,4-Tetrahydro-β-carboline-3-carboxylic acid, diethyl ether and L-Glutamine was higher in non-livebirth group. Limitations, reasons for caution These results can not be generalized to the entire population due to small sample size of our study. More work is needed on the subject. Wider implications of the findings We detected significant differences in some of the metabolites in patients with different reproductive outcomes. The metabolomic analysis of blastocyst spent culture medium can be used as an adjunt method for improving non invasive embryo evaluation in clinical practice after evidence based results. Trial registration number none

Postpartum readmission risk: a comparison between stillbirths and live births

Stillbirth occurs more commonly among pregnant people with comorbid conditions and obstetrical complications. Stillbirth also independently increases maternal morbidity and imparts a psychosocial hazard when compared with live birth. These distinct needs and burden may increase the risk for postpartum readmission after stillbirth. This study aimed to examine the risk for maternal postpartum readmission after stillbirth in comparison with live birth and to identify indications for readmission and the associated risk factors. This was a retrospective cohort of patients with singleton stillbirths or live births, delivered at ≥20 weeks' gestation, who were identified from the 2019 Nationwide Readmissions Database. The primary outcome was all-cause readmission within 6 weeks of discharge from the childbirth hospitalization. The association between stillbirth (vs live birth) and risk for readmission was assessed using multivariable regression models with adjustment for maternal age, sociodemographic characteristics, maternal and obstetrical conditions, and delivery characteristics. Within the stillbirth group, risk factors for readmission were further examined using multivariable regression. The secondary outcomes included principal indication for readmission (categorized based on principal diagnosis code of the readmission hospitalization) and timing of readmission (number of weeks after childbirth hospitalization). Differences in these secondary outcomes were compared between the stillbirth and live birth groups using chi-square tests. All analyses accounted for the complex sample design to generate nationally representative estimates. Postpartum readmission occurred in 2.7% of 16,636 patients with stillbirths, whereas it occurred in 1.6% of 2,870,677 patients with live births (unadjusted risk ratio, 1.65; 95% confidence interval, 1.47-1.86). The higher risk for readmission after stillbirth (vs live birth) persisted after adjusting for maternal, obstetrical, and delivery characteristics (adjusted risk ratio, 1.27; 95% confidence interval, 1.11-1.46). The distribution of principal indication for readmission differed after stillbirth and after live birth and included hypertension (30.2% vs 39.5%; unadjusted risk ratio, 0.76; 95% confidence interval, 0.63-0.93), mental health or substance use disorders (6.8% vs 3.6%; unadjusted risk ratio, 1.90; 95% confidence interval, 1.15-3.16), and venous thromboembolism (5.8% vs 2.0%; unadjusted risk ratio, 2.87; 95% confidence interval, 1.60-5.17). Among patients with stillbirths, 56.0% of readmissions occurred within 1 week, 71.8% within 2 weeks, and 88.1% within 4 weeks; the timing of readmission did not differ significantly between the stillbirth and live birth cohorts. Pregestational diabetes (adjusted risk ratio, 1.87; 95% confidence interval, 1.20-2.93), gestational diabetes (adjusted risk ratio, 1.67; 95% confidence interval, 1.03-2.71), hypertensive disorders of pregnancy (adjusted risk ratio, 1.80; 95% confidence interval, 1.31-2.47), obesity (adjusted risk ratio, 1.46; 95% confidence interval, 1.01-2.12), and primary cesarean delivery (adjusted risk ratio, 1.74; 95% confidence interval, 1.17-2.58) were associated with a higher risk for readmission after stillbirth, whereas higher household income was associated with a lower risk for readmission (eg, adjusted risk ratio for income ≥$82,000 vs $1-$47,999, 0.48; 95% confidence interval, 0.30-0.77). When compared with live births, the risk for postpartum readmission was higher after stillbirths, even after adjustment for differences in the patient demographic and clinical characteristics. Readmission for mental health or substance use disorders and venous thromboembolism is more common after stillbirths than after live births.

Associations between the artificial intelligence scoring system and live birth outcomes in preimplantation genetic testing for aneuploidy cycles

BackgroundSeveral studies have demonstrated that iDAScore is more accurate in predicting pregnancy outcomes in cycles without preimplantation genetic testing for aneuploidy (PGT-A) compared to KIDScore and the Gardner criteria. However, the effectiveness of iDAScore in cycles with PGT-A has not been thoroughly investigated. Therefore, this study aims to assess the association between artificial intelligence (AI)-based iDAScore (version 1.0) and pregnancy outcomes in single-embryo transfer (SET) cycles with PGT-A.MethodsThis retrospective study was approved by the Institutional Review Board of Chung Sun Medical University, Taichung, Taiwan. Patients undergoing SET cycles (n = 482) following PGT-A at a single reproductive center between January 2017 and June 2021. The blastocyst morphology and morphokinetics of all embryos were evaluated using a time-lapse system. The blastocysts were ranked based on the scores generated by iDAScore, which were defined as AI scores, or by KIDScore D5 (version 3.2) following the manufacturer’s protocols. A single blastocyst without aneuploidy was transferred after examining the embryonic ploidy status using a next-generation sequencing-based PGT-A platform. Logistic regression analysis with generalized estimating equations was conducted to assess whether AI scores are associated with the probability of live birth (LB) while considering confounding factors.ResultsLogistic regression analysis revealed that AI score was significantly associated with LB probability (adjusted odds ratio [OR] = 2.037, 95% confidence interval [CI]: 1.632–2.542) when pulsatility index (PI) level and types of chromosomal abnormalities were controlled. Blastocysts were divided into quartiles in accordance with their AI score (group 1: 3.0–7.8; group 2: 7.9–8.6; group 3: 8.7–8.9; and group 4: 9.0–9.5). Group 1 had a lower LB rate (34.6% vs. 59.8–72.3%) and a higher rate of pregnancy loss (26% vs. 4.7–8.9%) compared with the other groups (p < 0.05). The receiver operating characteristic curve analysis verified that the iDAScore had a significant but limited ability to predict LB (area under the curve [AUC] = 0.64); this ability was significantly weaker than that of the combination of iDAScore, type of chromosomal abnormalities, and PI level (AUC = 0.67). In the comparison of the LB groups with the non-LB groups, the AI scores were significantly lower in the non-LB groups, both for euploid (median: 8.6 vs. 8.8) and mosaic (median: 8.0 vs. 8.6) SETs.ConclusionsAlthough its predictive ability can be further enhanced, the AI score was significantly associated with LB probability in SET cycles. Euploid or mosaic blastocysts with low AI scores (≤ 7.8) were associated with a lower LB rate, indicating the potential of this annotation-free AI system as a decision-support tool for deselecting embryos with poor pregnancy outcomes following PGT-A.

Pregravid preparation of women with chronic endometritis in IVF cycles.

Aim: of our study was to improve the pregravid preparation of women with chronic endometritis to develop individual approaches to overcoming infertility, taking into account the state of endometrium. Materials and Methods: The study included 90 women (main group, n=90), 28 to 38 years with an anatomically normal uterus and chronic endometritis (CE). Patients were divided into 2 groups: group I - 45 women with CE who received conventional treatment; group II - 45 women with CE who received pregravid preparation by subendometrial injections of Platelet-Rich Plasma (PRP). Results: At the first stage of study, the ART statistical reports from 2015 to 2022 were analyzed at the Medical Center of Reproductive Health ≪Damia≫, (Ivano-Frankivsk). Analysis of the vaginal flora parameters before treatment at the first stage revealed the presence of conditionally pathogenic flora in culture from the cervical canal (Candida albicans - 2.4%, Escherihia coli - 4.8%, Staphylococcus epidermidis - 6.2% Enterococcus faecalis - 6.9%), and was evidence of a possible recurrence of inflammation during gestation. In the age category, the groups of patients were homogeneous, with no significant differences by the level of AMH and the level of CD 138. Biochemical pregnancy be present in 20 patients (44.4%) of group I and 28 (62.2%) of group II. Fertility within a year after the end of therapy was restored with the proposed method of therapy in most women (51.1%), in the comparison group this number was 11.1% lower. Pregnancy rate between the groups (I and II) did not differ significantly. The number of live births in group II - 19 births (42.2%) - was 2 times higher than I group (9 (20.0%), P<0.05). The most common complication for women in the comparison groups was early pregnancy loss. Among 18 (40.0%) clinical pregnancies of group I, 8 women (17.8%) had early miscarriage, 1 ectopic pregnancy (2.2%), while in group II clinical pregnancy be present in 23 women (51.1%). The number of terminated pregnancies was two times lower than in the first group (8.9% vs. 17.8%, P<0.05). Conclusions: Chronic endometritis is one of the main causes of pregnancy loss after in vitro fertilization. Patients of the second group were treated with the proposed method of subendometrial injections with Platelet-Rich Plasma (PRP), prepared from autologous blood, is an effective method of preparing the endometrium for embryo transfer and can increase the number of live births in patients with chronic endometritis.

A combined clinical and specific genes’ model to predict live birth for in vitro fertilization and embryo transfer patients

BackgroundWe aimed to develop an accurate model to predict live birth for patients receiving in vitro fertilization and embryo transfer (IVF-ET) treatment.MethodsThis is a prospective nested case–control study. Women aged between 18 and 38 years, whose body mass index (BMI) were between the range of 18.5–24 kg/m2, who had an endometrium of ≥ 8 mm at the thickest were enrolled from 2018/9 to 2020/8. All patients received IVF-ET treatment and were followed up until Jan. 2022 when they had reproductive outcomes. Endometrial samples during the window of implantation (LH + 6 to 9 days) were subjected to analyze specific endometrial receptivity genes’ expression using real-time PCR (RT-PCR). Patients were divided into live birth group and non-live birth group based on IVF-ET outcomes. Clinical signatures relevant to live birth were collected, analyzed, and used to establish a predictive model for live birth by univariate analysis (clinical model). Specific endometrial receptivity genes’ expression was analyzed, selected, and used to construct a predictive model for live birth by The Least Absolute Shrinkage and Selection Operator (LASSO) analysis (gene model). Finally, significant clinical factors and genes were used to construct a combined model for predicting live birth using multivariate logistical regression (combined model). Different models’ Area Under Curve (AUC) were compared to identify the most predictive model.ResultsThirty-nine patients were enrolled in the study, twenty-four patients had live births, fifteen did not. In univariate analysis, the odds of live birth for women with ovulation dysfunction was 4 times higher than that for women with other IVF-ET indications (OR = 4.0, 95% CI: 1.125 − 8.910, P = 0.018). Age, body mass index, duration of infertility, primary infertility, repeated implantation failure, antral follicle counting, ovarian sensitivity index, anti-Mullerian hormone, controlled ovarian hyperstimulation protocol and duration, total dose of FSH/hMG, number of oocytes retrieved, regiment of endometrial preparation, endometrium thickness before embryo transfer, type of embryo transferred were not associated with live birth (P > 0.05). Only ovulation dysfunction was used to construct the clinical model and its AUC was 0.688. In lasso analysis, GAST, GPX3, THBS2 were found to promote the risk of live birth. AUCs for GAST, GPX3, THBS2 reached to 0.736, 0.672, and 0.678, respectively. The gene model was established based on these three genes and its AUC was 0.772. Ovulation dysfunction, GAST, GPX3, and THBS2 were finally used to construct the combined model, reaching the highest AUC (AUC = 0.842).ConclusionsCompared to the single model, the combined model of clinical (Ovulation dysfunction) and specific genes (GAST, GPX3, THBS2) was more accurate to predict live birth for IVF-ET patients.

Biomarkers at 6 weeks' gestation in the prediction of early miscarriage in pregnancy following assisted reproductive technology.

Miscarriage is a major concern in early pregnancy among women having conceived with assisted reproductive treatments. This study aimed to examine potential miscarriage-related biophysical and biochemical markers at 6 weeks' gestation among women with confirmed clinical pregnancy following in vitro fertilization (IVF)/embryo transfer (ET) and evaluate the performance of a model combining maternal factors, biophysical and biochemical markers at 6 weeks' gestation in the prediction of first trimester miscarriage among singleton pregnancies following IVF/ET. A prospective cohort study was conducted in a teaching hospital between December 2017 and January 2020 including women who conceived through IVF/ET. Maternal mean arterial pressure, ultrasound markers including mean gestational sac diameter, fetal heart activity, crown rump length and mean uterine artery pulsatility index (mUTPI) and biochemical biomarkers including maternal serum soluble fms-like tyrosine kinase-1 (sFlt-1), placental growth factor (PlGF), kisspeptin and glycodelin-A were measured at 6 weeks' gestation. Logistic regression analysis was carried out to determine significant predictors of miscarriage prior to 13 weeks' gestation and performance of screening was estimated by receiver-operating characteristics curve analysis. Among 169 included pregnancies, 145 (85.8%) pregnancies progressed to beyond 13 weeks' gestation and had live births whereas 24 (14.2%) pregnancies resulted in a miscarriage during the first trimester. In the miscarriage group, compared to the live birth group, maternal age, body mass index, and mean arterial pressure were significantly increased; mean gestational sac diameter, crown rump length, mUTPI, serum sFlt-1, glycodelin-A, and the rate of positive fetal heart activity were significantly decreased, while no significant differences were detected in PlGF and kisspeptin. Significant prediction for miscarriage before 13 weeks' gestation was provided by maternal age, fetal heart activity, mUTPI, and serum glycodelin-A. The combination of maternal age, ultrasound (fetal heart activity and mUTPI), and biochemical (glycodelin-A) markers achieved the highest area under the curve (AUC: 0.918, 95% CI 0.866-0.955), with estimated detection rates of 54.2% and 70.8% for miscarriage before 13 weeks' gestation, at fixed false positive rates of 5% and 10%, respectively. A combination of maternal age, fetal heart activity, mUTPI, and serum glycodelin-A at 6 weeks' gestation could effectively identify IVF/ET pregnancies at risk of first trimester miscarriage.

Non-coding RNAs from seminal plasma extracellular vesicles and success of live birth among couples undergoing fertility treatment.

Background: Infertility remains a global health problem with male-factor infertility accounting for around 50% of cases. Understanding the molecular markers for the male contribution of live birth success has been limited. Here, we evaluated the expression levels of seminal plasma extracellular vesicle (spEV) non-coding RNAs (ncRNAs) in men of couples in relation with those with and without a successful live birth after infertility treatment. Method: Sperm-free spEV small RNA profiles were generated from 91 semen samples collected from male participants of couples undergoing assisted reproductive technology (ART) treatment. Couples were classified into two groups based on successful live birth (yes, n = 28) and (no, n = 63). Mapping of reads to human transcriptomes followed the order: miRNA > tRNA > piRNA > rRNA> "other" RNA > circRNA > lncRNA. Differential expression analysis of biotype-specific normalized read counts between groups were assessed using EdgeR (FDR<0.05). Result: We found a total of 12 differentially expressed spEV ncRNAs which included 10 circRNAs and two piRNAs between the live birth groups. Most (n = 8) of the identified circRNAs were downregulated in the no live birth group and targeted genes related to ontology terms such as negative reproductive system and head development, tissue morphogenesis, embryo development ending in birth or egg hatching, and vesicle-mediated transport. The differentially upregulated piRNAs overlapped with genomic regions including coding PID1 genes previously known to play a role in mitochondrion morphogenesis, signal transduction and cellular proliferation. Conclusion: This study identified novel ncRNAs profiles of spEVs differentiating men of couples with and without live birth and emphasizes the role of the male partner for ART success.

O-262 The vaginal and faecal microbiome in women with recurrent pregnancy loss (RPL) before pregnancy according to the reproductive outcome after referral

Abstract Study question Is the vaginal or faecal microbiome different between primary RPL (pRPL) and secondary RPL (sRPL) patients, and is it related to reproductive outcome after referral? Summary answer Before pregnancy, the vaginal microbiome differed between pRPL and sRPL, and the faecal microbiome was altered in those who did not achieve pregnancy after follow-up. What is known already RPL is a heterogeneous condition leaving 50% of the couples without any known risk factors after the initial diagnostic workup. The microbiome of the reproductive tract seems to be an essential factor in women’s health, including pregnancy loss. Only a few studies have investigated the vaginal microbiome in women with RPL and found dysbiosis with a decrease in Lactobacillus crispatus. To our knowledge, the faecal microbiome in women with RPL has never been investigated. Study design, size, duration A prospective cohort study including 106 women referred with unexplained RPL between 04/2018 and 12/2019. Patients were routinely screened for established risk factors as recommended in the ESHRE RPL guideline. Exclusion criteria were &amp;gt;40 years, &amp;gt;1 shared child, the use of antibiotics, antimycotics and antiviral medication within the past two weeks, known chromosomal aberrations and major uterine malformations. Follow-up ranged between 12-31 months. Participants/materials, setting, methods The women were referred with a minimum of three unexplained consecutive pregnancy losses (64 pRPL and 42 sRPL) to the tertiary Recurrent Pregnancy Loss Unit at Copenhagen University Hospital (Rigshospitalet and Hvidovre Hospital), Denmark. Vaginal and faecal samples were collected before pregnancy and shot-gun sequenced on a DNBSEQ-G400 sequencer (MGI) using the high-throughput sequencing set (PE150 1000016952; MGI). Main results and the role of chance The beta diversity in the vaginal samples was significantly different (p = 0.001) between pRPL and sRPL, with Lactobacillus crispatus dominating more women with pRPL compared to sRPL who were dominated by Lactobacillus iners. Overall, twenty-nine patients (27.3% of the cohort) had vaginal dysbiosis defined as &amp;lt; 60% Lactobacillus spp. with no significant difference between pRPL and sRPL (vaginal dysbiosis in 21.8% vs. 35.7%, p = 0.118). During follow-up, 93 (87.7%) patients achieved pregnancy of which 50 (53.8%) resulted in a live birth and 43 (46.2%) in another pregnancy loss. There were no differences in age, BMI, alpha or beta diversity in the vaginal samples between the live birth group and pregnancy loss group. In the live birth group, 11 patients (22.0%) had vaginal dysbiosis compared with 32.1% of the rest of the cohort, p = 0.242. Ninety-three of 106 women collected a faecal sample at home after the first consultation and there was a significant difference in both alpha diversity (p = 0.014), beta diversity (p = 0.05) and richness (p = 0.001) between women who achieved pregnancy compared with those who did not conceive after follow-up. Limitations, reasons for caution Patients with recurrent pregnancy loss constitute a heterogenic population, which underlines the importance of subgroup comparisons such as pRPL and sRPL. Wider implications of the findings These findings underline the role of an altered vaginal and faecal microbiome as a potential risk factor for RPL. In-depth knowledge about the altered microbiome compositions in these patients can contribute to the generation of future treatment strategies and potentially improve patient care. Trial registration number not applicable

P-158 Effect of women body mass index on in vitro embryos development and effectiveness of «Assisted reproductive technologies»

Abstract Study question Does a woman's body mass index (BMI) affect the development of an embryo in vitro? Summary answer Evaluation of the development of embryos indicates that the overweight affects the development of the embryo and associated with a lower rate of live births. What is known already Excess weight and obesity - as a cause of infertility, occures against the background of existing metabolic disorders. Severe thinness and, like obesity, often lead to violations of the interval and reproductive functions of a woman. An increase in adipose tissue of more than 20% or more leads to dysfunction of the hypothalamic-pituitary-ovarian system. With obesity and emaciation of the body, a woman has various forms of menstrual disorders, the frequency of uterine bleeding and also endometrial pathology increases. Study design, size, duration This study included the retrospective analysis of Assisted Reproductive Technologies (ART) cycles using statistical data on the development of embryos in the Embryological laboratory, pregnancy and childbirth was carried out from 2016 to 2018. Morphological studies were carried out using Time-Lapse embryo imaging. Participants/materials, setting, methods In this study, morphological and biostatistical methods were used. The cohort of patients aged 20 to 45 years was divided into four groups: 1623 embryos from 243 underweight women; 19542 embryos from 2270 normal weight women (control group); 9701 embryos from 1069 overweight women; and 2668 embryos from 343 obese women. Main results and the role of chance In the group of underweight women the pregnancy rate is 46%, which is 3% less than in overweight and obese group of women, while the percentage of the control group is 47.5%. However, the results of live births in group of women with obesity is 7% less than in women of normal weight, which is 29 and 36 percent respectively. In addition, a comparative analysis of the number of fertilized eggs showed that at a normal BMI, 6.3 ± 0.4 oocytes were fertilized, while in the group of women with second-degree obesity only 4.2 ± 0.3 oocytes. Based on the results of the T-test, it can be concluded that these indicators have differences with a significance level of less than 0.05. Also, the results obtained indicate differences in the maturity of embryos at the blastocyst stage, where blastocysts of excellent quality matured in the control group of 3.4 ± 0.4 embryos, and in the experimental group with obese women 2.4 ± 0.1 embryos. Furthermore, the frequency of live births showed a relationship between the norm and high BMI. In the control group, the result was 0.7 ± 0.02, while in first-degree obesity, the result was 0.2 ± 0.06. Limitations, reasons for caution The main limitation of this study is a verity of women with different anamnesis, which were not taken into account. The grading of blastocysts is subjected to a human factor. The limitation of the Time-lapse technology is the inability to rotate the embryos making it very difficult to observe. Wider implications of the findings This method needs further investigations about limitations, such as the age of patients, type and heaviness of infertility, existing diseases and previous pregnancies in anamnesis. Trial registration number Not Applicable

P-244 Nucleation errors at four-cell stage reduces chances of live births in single euploid-blastocyst transfers

Abstract Study question What is the impact of nucleation errors (NE) and morphokinetic parameters on live birth rates in single euploid blastocyst transfers? Summary answer Nucleation errors (NE) at the four-cell stage was the only parameter associated with decreased live birth (LB) rates. What is known already Morphokinetic studies using time-lapse monitoring (TLM) systems have proposed algorithms to select embryos with the highest implantation potential. However, no single biomarker can reliably predict live-birth so far. Morphokinetics can deselect embryos for Preimplantation-Genetic-Testing for Aneuploidies (PGT-A). However, it is unclear whether morphokinetic markers can be used for selecting embryos with highest chance of LB among the euploid blastocysts. Performing morphokinetics enable detection of features such as nuclear errors (NE) which could be underdiagnosed with intermittent evaluation. It remains to be determined to which extend NE, next to morphokinetic parameters, are associated with LB potential of euploid blastocysts. Study design, size, duration Retrospective observational study including 383 single euploid blastocyst transfers in FET cycles between October 2017 to June 2021. These blastocysts were graded ≥BL3CC (Gardner criteria) and underwent TE biopsy for PGT-A by Next Generation Sequencing (NGS). Only ICSI with fresh autologous cycles using ejaculated sperm were considered. Outcomes of euploid blastocyst transfers were recorded and as pregnant (P): biochemical pregnancy (BP) (n = 17), clinical miscarriage (CM) (n = 35), live birth (LB) (n = 205); or not pregnant (NP) (n = 126). Participants/materials, setting, methods Patient baseline characteristics (Age, AMH, BMI) and infertility duration were considered. Morphokinetics were annotated with time-lapse-imaging (KID-score) for time (t) of PB2 (tPB2), tPNa (PN-appearance), tPNf (PN-fading), t2-t9, CP (compaction), tM (Morula), tSB (start blastulation), tB (blastocyst) and timings between each developmental event. Nuclear errors (NE) such as micronucleation, binucleation, multinucleation were annotated when present in at least one blastomere at two- and four-cell stages between 19.4-51.4 hours and 22.1-63.1 hours, respectively, post-insemination. Main results and the role of chance Median (IQR) values of Age and AMH of female partner, and infertility duration were similar among the P and NP groups. BMI was significantly higher in patients who miscarried with Median (IQR) values of 28.4kg/m2 (26.1-33.4) vs &amp;lt; 27.9kg/m2 in all other groups (P &amp;lt; 0.01). Significantly more blastocysts with type A inner cell mass quality resulted in LB (24.9%) than to CM (14.3%), BP (5.9%) and NP (13.5%) (P &amp;lt; 0.001). Concerning TLM morphokinetics evaluation, none of the timed parameters evaluated, neither the timings between the different morphokinetics events resulted in significant differences among the LB and the other P and NP groups. However, LB group had significantly less euploid embryos presenting NE at four-cell stage (7.8%) compared to CM (17.0%), BP (23.5%) and NP (17.5%) (P = 0.02). In a logistic regression model, NE when present at four-cell stage but not at two-cell stage, had a negative impact in LB rates (OR:0.39, CI: 0.2-0.72; P = 0.003). Limitations, reasons for caution The retrospective nature of the study and the fact that NE evaluation was performed manually based on TLM videos by a single observer. Wider implications of the findings NE-constituted embryos can give rise to euploid blastocysts and have an implantation potential. Since these embryos are associated with lower LB, it is relevant to include NE as an evaluation parameter to deprioritize multinucleated four-cell embryos for transfer, even when they result in euploid blastocysts. Trial registration number not applicable

P-520 Risk factors for early pregnancy loss in women undergoing in vitro fertilization-embryo transfer

Abstract Study question What are the risk factors for early pregnancy loss (EPL) in infertility patients undergoing in vitro fertilization-embryo transfer (IVF-ET) cycles? Summary answer The risk factors for EPL were maternal age, 14-day β-human chorionic gonadotrophin(β-hcg) levels and cycle type. What is known already Pregnancy obtained through assisted reproductive technology still has a high risk of spontaneous early pregnancy abortion, and the cause of which is unclear. It not only affects the rate of ongoing pregnancy and live birth, but also increases the psychological and economic burden of patients. At present, there have been many studies on the risk factors of early pregnancy loss in IVF/ICSI-ET at domestic and overseas, but the results are different, which may be related to the selection of different research objects, sample size and statistical methods. Study design, size, duration A retrospective study of 6107 embryo transfer cycles collected from January 2018 to December 2018 was performed. The cycles were divided into the EPL group (n = 987) and the live birth group (n = 5120) according to the cycle outcomes. Participants/materials, setting, methods Baseline data and clinically relevant indicators were compared between the two groups, including maternal age, infertility years, infertility type, body mass index (BMI), tubal factor, ovulatory dysfunction, endometriosis, male factor, antral follicle count (AFC), single gene related genetic disease, follicle-stimulating hormone (FSH), estradiol (E2), luteinizing hormone (LH), 14-day β-hcg levels, prolactin (PRL), progesterone (P), testosterone (T), anti-Müllerian hormone (AMH), cycle type and the number of transplanted embryos. Main results and the role of chance The EPL rate of the infertility patients undergoing fresh/frozen-thaw embryo transfer cycle was 16.16%. Univariate analysis showed that there were statistically significant differences between the two groups in maternal age, AFC, 14-day β-hcg levels, T, AMH, infertility years, cycle type, infertility types, chromosome disease and male factors (P &amp;lt; 0.05). The results of binary logistics regression analysis showed that maternal age, 14-day β-hcg levels and cycle type were related factors to EPL (P &amp;lt; 0.05). Limitations, reasons for caution This study lacked some baseline data, such as adverse pregnancy history, abortion history, and previous transplant failure history, and male factors such as BMI, age, semen results, smoking history. The information about fetal chromosomes was also missing. Wider implications of the findings Clinical prevention and treatment of risk factors can be targeted to improve pregnancy outcomes after transplantation. Trial registration number none

P-666 High embryo yield (and not oocyte yield) despite low ovarian stimulation dose correlates with cumulative livebirth rate in normal-high responders undergoing IVF/ICSI

Abstract Study question Does a good response i.e., high oocyte or embryo yield with low ovarian stimulation correlate with cumulative live birth outcome in IVF/ ICSI treatment? Summary answer High ovarian sensitivity to stimulation (embryos to gonadotropin dose ratio) positively predicts cumulative live birth in IVF, while oocytes to stimulation dose ratio does not. What is known already The conventional belief that the higher the number of eggs retrieved, the better for IVF outcome has been challenged. Oocyte yield fails to predict fresh-cycle live birth in presence of high ovarian response. Ovarian sensitivity index (OSI) which is an expression of inherent ovarian responsiveness to stimulation remained overlooked as a predictor of success in IVF. We explore whether attention should be on the oocyte or embryo yield as a function of ovarian stimulation (OSI) can predictor of positive outcomes in IVF. Study design, size, duration Retrospective analysis of 750 patients who underwent IVF/ ICSI treatment with ≤150 IU of gonadotropin in an antagonist protocol due to predicted normal or high response. This is a further analysis of a previously published study covering a period between 1 January 2016 to 31 Dec 2018. Participants/materials, setting, methods Multi-centre study within a group company using the same treatment protocols. Participants were normal or high responders of IVF/ ICSI, predicted poor responders (according to Bologna criteria) were excluded. All patients received ≤150 IU daily dose of gonadotropin (FSH) in an antagonist cycle. Cumulative LBR calculated. Univariate and multivariate (regression) analysis, adjusting for all significant variables were done to find the predictors of cumulative livebirth. Main results and the role of chance When the women who achieved a livebirth were compared with those who did not, there was no difference in the mean BMI (23.7 vs 23.6, p = 0.81), antral follicle counts (p = 0.33), Anti-mullerian hormone levels (p = 0,89), mean starting (149.5 vs 140.5 IU/ day, p = 0.16), and total gonadotropin doses (1360.7 vs 1342.8 IU, p = 0.62). Women’s age (33.5 vs 35.3 years, p &amp;lt; 0.0001) and the number of embryos transferred (p &amp;lt; 0.0001) had significantly correlation with LBRs. There was no difference in the mean number of oocyte (9.1 vs 8,6, p = 0.21) or oocyte yield per unit of gonadotropin (7.4 vs 6.7, p = 0.32).. The mean number of embryos (5.4 vs 4.1, p &amp;lt; 0.0001), transferable embryos (2.5 vs 1.9, p = 0.0006) and embryo yield per unit of gonadotropin (4.1 vs 3.2, p &amp;lt; 0.0001) were significantly higher in the livebirth group. After adjusting for woman’s age and the number of embryos transferred, the embryo yield (both total (p &amp;lt; 0.0001) and transferable (p = 0.01) embryos and embryo yield per gonadotropin unit (p = 0.0007) were predictors of cumulative livebirth whereas oocyte yield and oocyte number per unit of gonadotropin were not. The above findings remained the same when only IVF cycles excluding male infertility or only the cycles with a fixed 150 IU/d gonadotropin dose were analysed. Limitations, reasons for caution A moderate-sized study population. The poor responders have been excluded to minimise variations in the protocol and stimulation dose. Wider implications of the findings Our study is a proof of concept of ‘ovarian sensitivity index’ indicating the inherent ovarian responsiveness rather than the strength of stimulation determines the IVF/ICSI outcome. Embryo yield appeared to be a predictor of cumulative livebirth while oocyte yield is not, in normal and high responders of IVF. Trial registration number Not applicable

Exploring the internal exposome of seminal plasma with semen quality and live birth: A Pilot Study

Infertility is clinically defined as the inability to achieve pregnancy within 12 months of regular unprotected sexual intercourse and affects 15% of couples worldwide. Therefore, the identification of novel biomarkers that can accurately predict male reproductive health and couples’ reproductive success is of major public health significance. The objective of this pilot study is to test whether untargeted metabolomics is capable of discriminating reproductive outcomes and understand associations between the internal exposome of seminal plasma and the reproductive outcomes of semen quality and live birth among ten participants undergoing assisted reproductive technology (ART) in Springfield, MA. We hypothesize that seminal plasma offers a novel biological matrix by which untargeted metabolomics is able to discern male reproductive status and predict reproductive success. The internal exposome data was acquired using UHPLC-HR-MS on randomized seminal plasma samples at UNC at Chapel Hill. Unsupervised and supervised multivariate analyses were used to visualize the differentiation of phenotypic groups classified by men with normal or low semen quality based on World Health Organization guidelines as well as by successful ART: live birth or no live birth. Over 100 exogenous metabolites, including environmentally relevant metabolites, ingested food components, drugs and medications, and metabolites relevant to microbiome-xenobiotic interaction, were identified and annotated from the seminal plasma samples, through matching against the NC HHEAR hub in-house experimental standard library. Pathway enrichment analysis indicated that fatty acid biosynthesis and metabolism, vitamin A metabolism, and histidine metabolism were associated sperm quality; while pathways involving vitamin A metabolism, C21-steroid hormone biosynthesis and metabolism, arachidonic acid metabolism, and Omega-3 fatty acid metabolism distinguished live birth groups. Taken together, these pilot results suggest that seminal plasma is a novel matrix to study the influence of the internal exposome on reproductive health outcomes. Future research aims to increase the sample size to validate these findings.

Serum steroid metabolome on the day of oocyte retrieval in women with polycystic ovarian syndrome and its association with pregnancy outcome of in vitro fertilization

Steroid hormone level is a crucial factor affecting the outcomes of in vitro fertilization/intracytoplasmic sperm injection (IVF/ICSI). The purpose of this study was to evaluate serum steroid metabolome on the day of oocyte retrieval in women with polycystic ovarian syndrome (PCOS) and explore whether specific steroids can be potential indicators to improve the prediction of pregnancy outcomes in PCOS patients undergoing IVF/ICSI. In this study, the serum levels of 21 steroids in 89 women with PCOS and 73 control women without PCOS on the day of oocyte retrieval of the first IVF/ICSI treatment cycle were measured by liquid chromatography-tandem mass spectrometry (LC-MS/MS). All patients subsequently received good-quality embryo transfer, and the correlation between their steroid profiles and pregnancy outcomes of the first embryo transfer (ET) was retrospectively analyzed. We found PCOS patients had aberrant levels of 11 out of 21 steroid hormones compared to control individuals, with androgen steroid hormones being considerably enhanced. Enzyme activity evaluation indicated that PCOS women might have abnormal activity of CYP17A1, CYP21A2, CYP11B2, CYP19A1, HSD3B, HSD11B, and HSD17B. Additionally, the level of 18-hydroxycorticosterone (p = 0.014), corticosterone (p = 0.035), and 17-hydroxypregnenolone (p = 0.005) were markedly higher in live birth group than in non- live birth group for PCOS women following frozen embryo transfer (FET). Multiple logistic regressions indicated that 18-hydrocorticosterone and 17-hydroxypregnenolone were independently associated with live birth outcomes of PCOS women following FET. Receiver operating characteristic (ROC) curve analysis revealed that 0.595 ng/mL for 18-hydrocorticosterone level (AUC: 0.6936, p = 0.014).and 2.829 ng/mL for 17-hydroxypregnenolone level (AUC: 0.7215, p = 0.005) were the best cutoff values to predict live birth outcomes of PCOS. In conclusion, the blood steroid metabolome was closely related to the IVF/ICSI outcomes of PCOS patients. 18-hydroxycorticosterone and 17-hydroxypregnenolone might be potential indicators to predict pregnancy outcomes of PCOS undergoing IVF/ICSI treatment.

Serum Cu, Zn and IL-1β Levels May Predict Fetal Miscarriage Risk After IVF Cycles: A NestedCase-Control Study.

To explore the association between serum-related indicators (levels of inflammatory cytokines and essential trace elements) and miscarriage risk among infertile women undergoing assisted reproductive techniques (ART) on the 14th day after embryo transfer, and to develop and establish a multivariable algorithm model that might predict pregnancy outcome. According to a nestedcase-control study design, a total of 100 miscarriage cases and 100 live birth controls were included in this study, and women in both groups were infertile and have underwent in vitro fertilization (IVF). Pregnancy tests were performed and serum levels of five essential trace elements (vanadium (V), copper (Cu), zinc (Zn), selenium (Se) and molybdenum (Mo)) and five inflammatory cytokines (interleukin-1β (IL-1β), IL-6, IL-8, IL-10 and tumor necrosis factor-α (TNF-α)) of the participants were measured on the 14th day after embryo transfer. The serum levels of five inflammatory cytokines were determined by multiple magnetic bead enzyme immunity analyzer; and the serum concentrations of five elements were determined simultaneously by inductively coupled plasma‒mass spectrometry (ICP ‒ MS). The logistic regression was used to evaluate the relationship between these serum indices and miscarriage risk among women undergoing ART, and a predictive model of pregnancy outcome based on these indices was established. The levels of IL-10, IL-1β and TNF-α of infertile women in the live birth group were significantly higher than those in the miscarriage group (p = 0.009, p < 0.001, p = 0.006), and the levels of V, Cu, Zn and Se of infertile women in the live birth group were also significantly higher than those in the miscarriage group (all p < 0.001). Through logistic regression analyses, we found that serum levels of IL-1β, TNF-α, V, Cu, Zn and Se were significantly and negatively associated with miscarriage risk. Different combination prediction models were generated according to the results of logistic regression analyses, and the combination of IL-1β, Cu and Zn had the best prediction performance. The area under the curve (AUC) was 0.776, the sensitivity of the model was 60% and the specificity was 84%. In conclusion,the serum-related indicators of women undergoing ART on the 14th day after embryo transfer, including the inflammatory cytokines such as IL-1β and TNF-α and the essential trace metal elements such as V, Cu, Zn and Se, were negatively correlated with miscarriage risk. A multivariate algorithm model to predict pregnancy outcome among women undergoing ART was established, which showed that IL-1β, Cu and Zn might synergistically predict pregnancy outcome.

The importance of infertility duration and follicle size according to pregnancy success in women undergoing ovulation induction with gonadotropins and intrauterine insemination

In this study, we aimed to evaluate the effect of infertility duration and dominant follicle size measured on the day of human chorionic gonadotropin (HCG) administration on pregnancy rates in infertile women undergoing ovulation induction (OI) with gonadotropin (GND). A total of 352 patients aged 20 to 41 years who were diagnosed with unexplained infertility or polycystic ovary syndrome (PCOS) were included in this study. Patients with a history of multifollicular development or follicle stimulating hormone (FSH) value more than 12 IU/ml were excluded from the study. The demographic and clinical features of the patients were obtained from the patients’ files and hospital automation system and recorded for each woman. The demographic and clinical features of the patients were recorded. Patients were divided into two groups as live birth (group 1, n = 47) and non-live birth groups (group 2, n = 305). There were no statistically significant differences in regard to age, infertility type, follicle stimulating hormone (FSH) level, oestradiol (E2) level, antral follicle count (AFC), cycle characteristics, GND type, number of follicles, E2 level and endometrial thickness on HCG day, total GND dose, dominant follicle size (p > 0.05). Infertility duration in group 1 was 3.5 ± 2.1; in group 2, 4.7 ± 3.9 years. This difference was statistically significant (p = .014). According to this study, live birth rates after GND + IUIs(intrauterine insemination) were significantly affected by the duration of infertility. But the dominant follicle size and endometrial thickness measured on HCG day in GND and IUI cycles did not have a significant effect on pregnancy rates. IMPACT STATEMENT What is already known on the subject? It is thought that the timing of the ovarian triggering is vital for the success of intrauterine insemmination (IUI) treatment. What do the results of this study add? According to our results live birth rates after GND + IUIs were significantly affected by the duration of infertility. What are the implications of these findings for clinical practice and/or further research? It is determined that the duration of infertility is significant and patients should be encouraged to the treatment as soon as possible.

Influencing factors and predictive model of live birth involving low-grade blastocyst frozen–thawed transfer: a retrospective study

BackgroundWhether only low-grade blastocysts should undergo freeze–thaw transfer during the in vitro fertilization/intracytoplasmic sperm injection cycle remains controversial; however, high-quality embryos cannot be obtained from some patients. Therefore, we aimed to identify factors that may affect the live birth.MethodsA total of 662 couples with only low-grade blastocysts who voluntarily accepted freeze–thaw blastocyst transfer at a single reproductive center over a 7-year period were followed-up. According to the outcome after transfer, they were divided into live birth group and failed pregnancy group. A nomogram was constructed for predicting live births.ResultsBaseline information and clinical treatment characteristics of patients in the two groups were comparable. Fifty-two of the 662 cycles (7.9%) resulted in live birth. Paternal age, maternal basal luteinizing hormone level, endometrial preparation scheme, and blastocyst development days were independent factors that affected low-grade blastocyst freeze–thaw transfer outcomes. The predictive model constructed based on these four factors presented favorable calibration and discriminatory abilities (area under the curve, 0.734; 95% confidence interval, 0.781–0.813).ConclusionsFor patients who exclusively underwent low-grade blastocyst freeze–thaw transfer, advanced paternal age and a high level of maternal basal luteinizing hormone adversely affected low-grade blastocyst freeze–thaw transfer outcomes. Artificial cycle preparation of the endometrium and day 5 blastocyst selection may improve the probability of live birth.

Can Anti-Müllerian Hormone levels predict future pregnancy outcomes in recurrent pregnancy loss?

ObjectiveSerum Anti-Müllerian Hormone (AMH) levels have been shown to be lower among women who have experienced recurrent pregnancy loss (RPL) compared with the general population. However, it is unclear whether it can predict livebirth. This study aims to determine whether AMH can predict the likelihood of a livebirth in women with RPL. Study DesignProspective analysis of a consecutive cohort of women undergoing investigation for RPL in a tertiary referral centre over a seven year period (August 2014 -December 2021). Analysis was performed using descriptive statistics, chi-square models and logistic regression models adjusting for maternal age and previous livebirth. Exclusion criteria for the regression analysis included abnormal parental karyotype and abnormal pelvic ultrasound scan. Pregnancy outcome was defined as livebirth or further pregnancy loss. ResultsThere were 488 women who underwent investigation of RPL during the study period. Of these, 65.2% (n = 318) conceived following attendance at the clinic. The majority of these women (69.4%, n = 221) proceeded to have a livebirth. There were no differences in median AMH levels between the livebirth group and the further pregnancy loss group (11 pmol/L vs 9 pmol/L respectively (p = 0.083). AMH did not affect clinical pregnancy rates (p = 0.77, 95% CI = 0.99 [0.98, 1.01]) or pregnancy outcome (p = 0.30, 95% CI = 1.01 [0.99, 1.04]). Abnormal pelvic ultrasonography (p = 0.04) and an abnormal parental karyotype (p = 0.04) were associated with an increased likelihood of a subsequent pregnancy loss. ConclusionAlthough AMH levels may have some utility in counselling of some couples with RPL, these contemporaneous data indicate that low AMH does not negatively influence subsequent pregnancy outcome in women with recurrent pregnancy loss.