Demographic distributions and clinical results of assisted reproduction techniques in Turkey in 2019: a descriptive survey.

Objective: The aim of this study was to describe characteristics and outcomes of assisted reproductive technology (ART) cycles performed in 2019 in Turkey. Material and Methods: One-hundred and sixty-five ART centers in Turkey were invited to submit data. The survey was sent to center directors via e-mail with anonymous links by Qualtrics™. The survey involved questions about their patient characteristics, clinical practices, and outcomes. Results: Forty-one (24.8%) centers responded to e-mails, and data gathered from 25 centers was included in the analyses. In 25 centers, 18,127 fresh or frozen transfers were carried out during the study period, of which 7796 (43.0%) were fresh and the rest were either frozen (45.2%) or embryo transfers (ET) with preimplantation genetic testing (PGT) (11.8%). The live birth rate per ET was as 30.6%, 40.1%, and 50.7% in fresh, frozen and PGT cycles, respectively. A single embryo was transferred in 65.3% of all transfers and singleton live births comprised 86.1% of all deliveries. For cycles with intrauterine insemination, 1407 were started in 2019, and 195 clinical pregnancies, 150 live births with 19 multiple pregnancies occurred. A total of 1513 ART cycles were initiated for foreign patients. Russia (29.6%), Germany (7.4%), Iraq (4.6%), Uzbekistan (3.1%), and Syria (1.4%) were the top five countries with most patients coming to Turkey for ART. Conclusion: The survey results are in parallel with the reports of international institutions and organizations. With repeated editions, the data collected with annual surveys can be used to inform ART practices in the coming years.

Subcutaneous progesterone administration provides a similar ongoing pregnancy rate compared with intramuscular progesterone administration in hormone replacement therapy frozen embryo transfer cycles.

Objective: To compare the ongoing pregnancy rates (OPRs) for subcutaneous progesterone (SC-P) to intramuscular progesterone (IM-P) in hormone replacement therapy used in frozen embryo transfer (FET) cycles. Design: Prospective nonrandomized cohort study. Setting: Private fertility clinic. Patients: The study enrolled 224 patients scheduled for hormone replacement therapy (HRT)-FET cycles with SC-P (n = 133) or IM-P (n = 91). The route of P administration was decided according to the patient's preference and accessibility to the hospital. In the first FET cycle of a freeze-all cycle using single blastocyst transfers, a woman aged ≤35 was included. Main Outcomes: Ongoing pregnancy (OP). Results: The demographic, cycle, and embryologic characteristics were similar between groups. The clinical pregnancy rates (86/133[64.7%] vs. 57/91[62.6%]); miscarriage rates (21/86 [24.4%] vs. 10/57 [17.5%]), and OPR (65/133 [48.9%] vs. 47/91 [51.6%]) were comparable between the SC-P and IM-P groups. Binary logistic regression for OP as the dependent factor revealed that blastocyst morphology was found to be a significant independent prognosticator (for poor quality embryos adjusted odds ratio, 0.11; 95% confidence interval, 0.029-0.427) and progesterone route (SC-P vs. IM-P) was an insignificant prognosticator (adjusted odds ratio, 0.694; 95% confidence interval, 0.354-1.358). Conclusions: The OPR for SC-P administration was similar to that for IM-P in HRT-FET cycles. The effect of ET-day P levels may vary regarding the administration route. Randomized controlled trials comparing different P administration routes are needed, and large-scale prospective trials are warranted to evaluate the ET-day P levels on pregnancy outcome.

The number of prior pregnancy losses does not impact euploidy rates in young patients with idiopathic recurrent pregnancy loss.

PURPOSE: Our study aimed to determine the possible factors that might impact the probability of obtaining a euploid blastocyst following intracytoplasmic sperm injection (ICSI) and preimplantation genetic testing for aneuploidy (PGT-A) procedures in idiopathic recurrent pregnancy loss (RPL) patients. METHODS: This single-center retrospective cohort analysis included 180 oocyte retrieval cycles of 166 women under 35 years old and those diagnosed with idiopathic RPL according to American Society of Reproductive Medicine (ASRM) guidelines. Trophectoderm biopsy and next-generation sequencing (NGS) were the techniques used. Patients were stratified by the number of previous losses (Group A: 2, Group B: 3, and Group C: > 3). RESULTS: Baseline and embryological characteristics showed no statistically significant differences. The euploidy rate per analyzed blastocyst was comparable within the groups (63.3%, 58.2%, and 58.5%; p = 0.477). Logistic regression analyses confirmed that only the trophectoderm scores of A and B increased the probability of obtaining a euploid embryo [OR: 1.82, 95% CI (1.120-2.956), p: 0.016]. CONCLUSION: It is concluded that there was no correlation between the number of previous losses and the chance of finding at least one euploid embryo in ICSI cycles of women younger than 35 years.

Impact of trophoectoderm biopsy for preimplantation genetic testing on serum ß-hCG levels, time of delivery and birthweight following frozen embryo transfer cycles.

Aim: This study investigated whether trophoectoderm (TE) biopsy adversely impacts serum ß-human chorionic gonadotropin (hCG) level on the 15th day of embryo transfer (ET), delivery week and birthweight, between biopsied and unbiopsied embryo groups, in a cohort of women who delivered a singleton baby, following frozen-thawed ET.Methods: All women having had a live birth after blastocyst ETs following frozen ET cycles with preimplantation genetic testing (PGT) were included. A control group was selected among women who had a live birth following single frozen blastocyst transfer without PGT-A at the same period in our clinicResults: One hundred fifteen and 173 cycles with- and without-PGT, respectively, were included. Serum ß-hCG level on the 15th day after ET was comparable between the groups (p = .336). Average birthweight of the babies born following biopsied embryos were significantly lower (3200 vs. 3380; p = .027). Women who received trophectoderm biopsied embryos had a significantly higher probability of having a baby weighing ≤1500 g and 1500-2500 g (p = .022) or ≤2500 g (p = .008). Proportion of preterm delivery was significantly higher in the biopsy group (p = .023). However, after adjusting for potential covariates, trophectoderm biopsy did not seem to increase the risk of preterm birth (OR 1.525; 95% CI, 0,644-3.611; p = .338)Conclusions: TE biopsy does not seem to impact serum ß-hCG level on the 15th day after ET. Average birthweight is lower when a biopsied embryo was transferred. After adjusting for potential covariates, trophectoderm biopsy does not seem to increase the risk of preterm birth.

Does endometrial thickness affect birth weight and serum levels of pregnancy-associated plasma protein-A in frozen cycles?

OBJECTIVE: To investigate whether endometrial thickness (EMT) affects birth weight and maternal serum pregnancy-associated plasma protein-A (PAPP-A) level in singleton pregnancies following frozen-thawed blastocyst transfer (FET). METHODS: This retrospective study included women who underwent first-trimester combined screening for aneuploidy after FET, and subsequently delivered a singleton live birth after 24 weeks of gestation at a private in-vitro fertilization centre. The subjects were stratified into three groups based on EMT percentiles: <7.7 mm (<10th percentile), 7.7-12 mm (10-90th percentile), and > 12 mm (>90th percentile). Multi-variable linear regression analysis was undertaken to identify the association between birth weight and EMT after adjusting for variables with p < 0.1 on univariate analysis. RESULTS: In total, 560 women met the inclusion criteria. Mean (±standard deviation) birth weight was 3127 ± 783 g in those with EMT < 7.7 mm, 3225 ± 644 g in those with EMT 7.7-12 mm, and 3256 ± 599 g in those with EMT > 12 mm (p = 0.577). Mean PAPP-A and PAPP-A < 0.5 rates were similar in the three groups. On multi-variate analysis, maternal serum PAPP-A was found to be a significant predictor of birth weight (p = 0.013), but EMT was not found to be predictive of birth weight on univariate or multi-variate analysis. CONCLUSION: This study provided a new angle to evaluate the association between EMT and neonatal birth weight by analysing this association along with maternal serum PAPP-A as a marker for placental function, suggesting that EMT is not an independent factor for neonatal birth weight.

The use of copy number loads to designate mosaicism in blastocyst stage PGT-A cycles: fewer is better.

STUDY QUESTION: How well can whole chromosome copy number analysis from a single trophectoderm (TE) biopsy predict true mosaicism configurations in human blastocysts? SUMMARY ANSWER: When a single TE biopsy is tested, wide mosaicism thresholds (i.e. 20-80% of aneuploid cells) increase false positive calls compared to more stringent ones (i.e. 30-70% of aneuploid cells) without improving true detection rate, while binary classification (aneuploid/euploid) provides the highest diagnostic accuracy. WHAT IS KNOWN ALREADY: Next-generation sequencing-based technologies for preimplantation genetic testing for aneuploidies (PGT-A) allow the identification of intermediate chromosome copy number alterations potentially associated with chromosomal mosaicism in TE biopsies. Most validation studies are based on models mimicking mosaicism, e.g. mixtures of cell lines, and cannot be applied to the clinical interpretation of TE biopsy specimens. STUDY DESIGN, SIZE, DURATION: The accuracy of different mosaicism diagnostic thresholds was assessed by comparing chromosome copy numbers in multiple samples from each blastocyst. Enrolled embryos were donated for research between June 2019 and September 2020. The Institutional Review Board at the Near East University approved the study (project: YDU/2019/70-849). Embryos showing euploid/aneuploid mosaicism (n = 53), uniform chromosomal alterations (single or multiple) (n = 25), or uniform euploidy (n = 39) in their clinical TE biopsy were disaggregated into five portions: the inner cell mass (ICM) and four TE segments. Collectively, 585 samples from 117 embryos were analysed. PARTICIPANTS/MATERIALS, SETTING, METHODS: Donated blastocysts were warmed, allowed to re-expand, and disaggregated in TE portions and ICM. PGT-A analysis was performed using Ion ReproSeq PGS kit and Ion S5 sequencer (ThermoFisher). Sequencing data were blindly analysed with Ion Reporter software to estimate raw chromosome copy numbers. Intra-blastocyst comparison of copy number data was performed employing different thresholds commonly used for mosaicism detection. From copy number data, different case scenarios were created using more stringent (30-70%) or less stringent criteria (20-80%). Categorical variables were compared using the two-sample z test for proportions. MAIN RESULTS AND THE ROLE OF CHANCE: When all the five biopsies from the same embryo were analysed with 30-70% thresholds, only 8.4% (n = 14/166) of patterns abnormal in the original analysis revealed a true mosaic configuration, displaying evidence of reciprocal events (3.6%, n = 6/166) or confirmation in additional biopsies (4.8%, n = 8/166), while most mosaic results (87.3% of total predicted mosaic patterns) remained confined to a single TE specimen. Conversely, uniform whole chromosome aneuploidies (28.3% of total patterns, n = 47/166) were confirmed in all subsequent biopsies in 97.9% of cases (n = 46/47). When 20-80% thresholds were employed (instead of 30-70%), the overall mosaicism rate per biopsy increased from 20.2% (n = 114/565) to 40.2% (n = 227/565). However, the use of a wider threshold range did not contribute to the detection of additional true mosaic patterns, while significantly increasing false positive mosaic patterns from 57.8% to 79.5% (n = 96/166; 95% CI = 49.9-65.4 vs n = 271/341; 95% CI = 74.8-83.6, respectively) (P < 0.00001). Moreover, the shift of the aneuploid cut-off from 70% to 80% of aneuploid cells resulted in mosaicism overcalling in the high range (50-80% of aneuploid cells), impacting the accuracy of uniform aneuploid classification. Parametric analysis of thresholds, based on multifocal analysis, revealed that a binary classification scheme with a single cut-off at a 50% level provided the highest sensitivity and specificity rates. Further analysis on technical noise distribution at the chromosome level revealed a greater impact on smaller chromosomes. LIMITATIONS, REASONS FOR CAUTION: While enrolment of a population enriched in embryos showing intermediate chromosome copy numbers enhanced the evaluation of the mosaicism category compared with random sampling such study population selection is likely to lead to an overall underestimation of PGT-A accuracy compared to a general assessment of unselected clinical samples. This approach involved the analysis of aneuploidy chromosome copy number thresholds at the embryo level; future studies will need to evaluate these criteria in relation to clinical predictive values following embryo transfers for different PGT-A assays. Moreover, the study lacked genotyping-based confirmation analysis. Finally, aneuploid embryos with known meiotic partial deletion/duplication were not included. WIDER IMPLICATIONS OF THE FINDINGS: Current technologies can detect low-intermediate chromosome copy numbers in preimplantation embryos but their identification is poorly correlated with consistent propagation of the anomaly throughout the embryo or with negative clinical consequences when transferred. Therefore, when a single TE biopsy is analysed, diagnosis of chromosomal mosaicism should be evaluated carefully. Indeed, the use of wider mosaicism thresholds (i.e. 20-80%) should be avoided as it reduces the overall PGT-A diagnostic accuracy by increasing the risk of false positive mosaic classification and false negative aneuploid classification. From a clinical perspective, this approach has negative consequences for patients as it leads to the potential deselection of normal embryos for transfer. Moreover, a proportion of uniform aneuploid embryos may be inaccurately categorized as high-level mosaic, with a consequent negative outcome (i.e. miscarriage) when inadvertently selected for transfer. Clinical outcomes following PGT-A are maximized when a 50% threshold is employed as it offers the most accurate diagnostic approach. STUDY FUNDING/COMPETING INTEREST(S): The study was supported by Igenomix. The authors not employed by Igenomix have no conflicts of interest to declare. TRIAL REGISTRATION NUMBER: N/A.

Noninvasive amino acid turnover predicts human embryo aneuploidy.

Assisted reproduction technology has two significant problems: low success rates and multiple pregnancies. Because of these problems, the priority in IVF clinics is to develop a potential diagnostic test that can be used to select the embryos with the ultimate developmental competence. Aneuploidy screening as embryo selection criteria will ensure that the transferred embryos are euploid and high implantation rate. We hypothesize that aneuploidy in human preimplantation embryos could be discriminated by their amino acid metabolism profile in the spent culture media. Preimplantation genetic testing for aneuploidy results and spent embryo culture medium amino acid content were analyzed for 58 couples. The next-generation sequencing technique was used and coupled with TE biopsy. Forty euploid and 71 aneuploid blastocysts were evaluated. Embryos were cultured individually until day 5 or 6 of embryo development. Spent culture medium was collected after finishing the culture. There was no statistical difference between D3 and D5 embryo morphology between euploid and aneuploid embryos (p > .05). Eight amino acids, including SER, GLY, HIS, ARG, THR, ALA, PRO, and TYR, were detected in the culture medium from the blank control group, euploid group, and aneuploid group. Only TYR amino acid concentration was found significantly higher in the aneuploid group compared to the euploid group (p < .003). Tyrosine amino acid levels equal to and above 76.38 µmol/L could be considered aneuploid. Aneuploid embryos demonstrate altered amino acid turnover in vitro relative to euploid counterparts. A noninvasive method of amino acid profiling will be of value as a tool for routine preimplantation embryo selection among all patient groups.

Birth of a Healthy Baby 9 Years After a Surgically Successful Deceased Donor Uterus Transplant.

OBJECTIVE: To describe surgical procedures, previous failed pregnancies, methods for overcoming pregnancy failure and, most importantly, birth of a healthy infant, in a uterus transplantation from a deceased donor. BACKGROUND: Majority of uterus transplants have involved live donors, but several advantages make deceased donor transplantation a practicable option, principally by eliminating surgical risks to the live donor. METHODS: Uterus transplantation from a deceased donor was performed in September 2011 in Turkey. After 5 miscarriages, perfusion computed tomography revealed an obstructed blood-outflow. To overcome this blood flow obstruction, a saphenous vein graft was anastomosed between utero-ovarian and left ovarian vein with laparotomy. Follow-up computed tomography confirmed resolution of venous congestion and a decrease in uterine volume. RESULTS: Following vascular augmentation surgery, fetal cardiac activity were observed 28 days after the first embryo transfer attempt. Preterm premature rupture of the membranes was diagnosed at 19 weeks' gestation. Cesarean section was planned at 28 weeks' gestation due to intrauterine growth restriction and suspected preeclampsia. A healthy 760 g male baby was delivered. The baby was discharged from the neonatal intensive care unit 79 days after delivery in good condition weighing 2475 g. CONCLUSIONS: Deceased donor uterus transplantation is a reasonable approach for treating uterine factor-related infertility. In case of recurrent miscarriages, regional vascular augmentation by arterial or venous supercharging may be required to overcome regional misperfused regions determined by imaging studies.

Endometrial microbiota composition is associated with reproductive outcome in infertile patients.

BACKGROUND: Previous evidence indicates associations between the female reproductive tract microbiome composition and reproductive outcome in infertile patients undergoing assisted reproduction. We aimed to determine whether the endometrial microbiota composition is associated with reproductive outcomes of live birth, biochemical pregnancy, clinical miscarriage or no pregnancy. METHODS: Here, we present a multicentre prospective observational study using 16S rRNA gene sequencing to analyse endometrial fluid and biopsy samples before embryo transfer in a cohort of 342 infertile patients asymptomatic for infection undergoing assisted reproductive treatments. RESULTS: A dysbiotic endometrial microbiota profile composed of Atopobium, Bifidobacterium, Chryseobacterium, Gardnerella, Haemophilus, Klebsiella, Neisseria, Staphylococcus and Streptococcus was associated with unsuccessful outcomes. In contrast, Lactobacillus was consistently enriched in patients with live birth outcomes. CONCLUSIONS: Our findings indicate that endometrial microbiota composition before embryo transfer is a useful biomarker to predict reproductive outcome, offering an opportunity to further improve diagnosis and treatment strategies. Video Abstract.

Decision regret and associated factors following oocyte cryopreservation in patients with diminished ovarian reserve and/or age-related fertility decline.

PURPOSE: To evaluate the prevalence and factors associated with decision regret following oocyte cryopreservation (OC) in women with diminished ovarian reserve (DOR) and/or age-related fertility decline (ARFD). METHODS: A cross-sectional survey study was conducted to five hundred fifty-two women with DOR and/or ARFD who underwent OC between 2014 and 2019 in two private-assisted reproductive units in Istanbul, Turkey. Decision regret was measured using the validated Decision Regret Scale (DRS). RESULTS: The median and mean DRS scores were 10 (interquartile range: 25) and 13.4 (SD: 13.2, range 0-70), respectively. Eighty-five (52.5%) women reported mild regret and 26 (16%) had moderate to severe regret. Decision regret was inversely associated with the belief in fate regarding childbearing and trust in the efficacy of OC. CONCLUSIONS: The prevalence of severe decision regret among patients with DOR and/or ARFD undergoing OC is low. Women who had belief in fate and trusted in the efficacy of oocyte cryopreservation had significantly lower decisional regret.

Which factors affect the likelihood of miscarriage after single euploid blastocyst transfer?

RESEARCH QUESTION: Which parameters affect the likelihood of miscarriage after single euploid frozen-thawed blastocyst transfer (FBT)? DESIGN: In this retrospective study, clinical and laboratory data from 1051 single euploid FBTs were evaluated. Exclusion criteria were endocrine or systemic pathologies, uterine anomalies or pathologies, unilateral or bilateral hydrosalpinx, karyotypic abnormalities (either maternal or paternal) or thrombophilia. Patients were divided into two groups according to pregnancy outcome: live birth and miscarriage. RESULTS: Body mass index (BMI) (25.98 ± 0.5 versus 24.36 ± 0.21, P = 0.019), duration of infertility (6.62 ± 0.54 versus 4.92 ± 0.18, P = 0.006) and number of previous miscarriages (1.36 ± 0.13 versus 0.79 ± 0.05, P < 0.001) were significantly higher in the miscarriage group (n = 100) than in the live birth group (n = 589). Although the trophectoderm and inner cell mass (ICM) percentage scores were not statistically different among the miscarriage and live birth groups, the percentage of day-6 biopsied embryos was significantly higher in the miscarriage group. Binary logistic regression analysis revealed that BMI (OR 1.083, 95% CI 1.013 to 1.158, P = 0.02) and number of previous miscarriages (OR 1.279, 95% CI 1.013 to 1.158, P = 0.038) were independent factors for miscarriage. Patients with elevated BMI and a higher number of miscarriages were at increased risk of miscarriage. CONCLUSION: After a single euploid FBT, BMI and number of previous miscarriages are predictors of miscarriage. Lifestyle interventions before FBT may decrease miscarriage rates.

Euploidy rates of embryos in young patients with good and low prognosis according to the POSEIDON criteria.

RESEARCH QUESTION: Does an association exist between ovarian reserve, ovarian response and embryonic euploidy in female patients under age 35 years? DESIGN: This was a retrospective analysis of intracytoplasmic sperm injection and preimplantation genetic testing for aneuploidies cycles among patients enrolled at Bahceci Fulya IVF Center between January 2016 and August 2019. A total of 133 patients in POSEIDON group 1 (suboptimal responder; female age <35 years, antral follicle count [AFC] ≥5, number of oocytes retrieved <10) (group A), 133 patients in POSEIDON group 3 (expected low responder; female age <35 years, AFC <5) (group B) and 323 in the non-low-prognosis group (female age <35 years, AFC ≥5 and number of oocytes retrieved >9) (group C) were included. RESULTS: There was no significant difference in euploidy rate per embryo among the three groups (61.7% [145/235] for group A versus 53.5% [68/127] for group B versus 62% [625/1008] for group C; P = 0.13). The cancellation rate in cycles without a euploid blastocyst was significantly lower in group C than groups A and B (8.4% versus 12.8% and 16.5%; P = 0.034). Multivariate regression analysis indicated that the ovarian response group did not significantly affect the probability of obtaining a euploid embryo. Trophectoderm score 'C' (odds ratio 0.520, P = 0.007) and inner cell mass score 'C' (odds ratio 0.480, P < 0.001) were associated with a decreased probability of obtaining a euploid embryo. CONCLUSIONS: These results confirm that POSEIDON group 1 and group 3 and non-low-prognosis patients have different probabilities of euploid embryos being obtained per cycle. However, euploidy rates per embryo are not affected by the patient's ovarian reserve and response.

The impact of patient, embryo, and translocation characteristics on the ploidy status of young couples undergoing preimplantation genetic testing for structural rearrangements (PGT-SR) by next generation sequencing (NGS).

PURPOSE: To evaluate the factors that affect the incidence of euploid balanced embryos and interchromosomal effect (ICE) in carriers of different structural rearrangements. METHODS: This retrospective study includes 95 couples with reciprocal translocations (RecT) and 36 couples with Robertsonian translocations (RobT) undergoing Preimplantation Genetic Testing for Structural Rearrangements (PGT-SR) between March 2016 and July 2019. Next-generation sequencing (NGS) was the technique used coupled with trophectoderm (TE) biopsy. Only cases with females under 38 years were included. A total of 532 blastocysts were evaluated. RESULTS: The euploidy rate was similar in RobT when compared with RecT carriers [57/156 (36.5%) vs. 112/376 (29.8%), p = 0.127]. The pure ICE rate was significantly higher in RobT carriers [48/156 (30.8%) vs. 53/376 (14.1%), p < 0.001] than it was in RecT carriers. Female age was the independent factor for the probability of obtaining a euploid embryo in RecT and RobT carriers, and increasing female age decreases the probability of obtaining a euploid embryo. In RecT carriers, no significant differences were observed in euploidy rates, pure ICE, or combined ICE according to the length of the translocated fragment and the chromosome group. However, total ICE was significantly lower when there was a breakpoint in the short chromosome arm together with a breakpoint in the long arm [(44/158 (27.8%) for pq or qp, 51/155 (32.9%) for pp and 30/63 (47.6%) for qq; p = 0.02]. CONCLUSION: The incidence of euploid/balanced blastocysts was similar in both types of translocations. However, there was a significant increase in pure ICE in RobT compared to RecT carriers. In RecT carriers, the presence of the breakpoints in the long arm of the chromosomes involved in the rearrangement resulted in a higher total ICE.

Being on the side of old findings: progesterone elevation on the day of oocyte maturation induction does not affect embryological parameters throughout the blastocyst culture period.

PURPOSE: To investigate whether there is any detrimental effect of progesterone elevation (PE) on the day of oocyte maturation induction on embryological development potentials. METHODS: This retrospective single-center cohort study included a total of 1485 individual intracytoplasmic sperm injection (ICSI) cycles between January 2014 and December 2018. Serum progesterone (P) levels were measured on the day of oocyte maturation induction following the GnRH antagonist suppression protocol. Embryological parameters such as maturation, fertilization rate (FR), top-quality embryo (TQE) formation rate per 2PN on day 3, and excellent-quality blastocyst (EQB) formation rate per 2PN on day 5/6 were recorded. The inclusion criteria for women were an age ≤ 37 years, a BMI ≤ 30 kg/m2, and access to a total sperm concentration ≥ 2 million. Groups were stratified according to the serum P levels using the cut-off levels of < 0.8 ng/ml; 0.8-1.49 ng/ml; and ≥ 1.5 ng/ml. RESULTS: Peak E2 level and total number of oocytes retrieved were significantly related to PE (p < 0.001). FR did not display a significance difference between groups (p = 0.108). The TQE and the blastulation rates were not affected by PE (p = 0.82 and p = 0.68, respectively). Chi square analysis revealed a significant relationship between PE and the EQB formation rate (p = 0.01). GEE analysis failed to present any statistical significance regarding the effect of PE on neither the TQE nor the EQB formation rates per 2PN [OR 1.07; 95% (0.98-1.16) p = 0.113 and OR 0.93; 95% (0.80-1.07) p = 0.32, respectively]. CONCLUSIONS: In accordance with previously published papers, our study could not find any detrimental effect of PE on embryological outcomes throughout the blastocyst culture period.

Impact of Isthmocele on Assisted Reproductive Treatment Outcomes: An Age-matched Retrospective Study.

STUDY OBJECTIVE: To evaluate whether the presence of an ultrasonographic visible isthmocele has an impact on the chance of pregnancy in women undergoing embryo transfer. DESIGN: Age-matched retrospective study. SETTING: Private-assisted reproductive center. PATIENTS: The study included a total of 150 patients with a history of cesarean delivery with 75 isthmocele cases and 75 controls. All patients underwent embryo transfer from January 2017 through June 2018. The diagnosis of isthmocele was based on transvaginal ultrasound assessment. Isthmocele was defined as an anechoic indentation on the previous cesarean scar at the midsagittal plane, with a depth of >1 mm. INTERVENTIONS: Embryo transfer. MEASUREMENTS AND MAIN RESULTS: The groups were similar in patient and treatment characteristics. The live birth rate was 44% in the isthmocele group and 46.7% in the control group (odds ratio [OR] 0.89; 95% confidence interval [CI], 0.47-1.71; p = .743). The clinical pregnancy rates were 49.3% and 50.7%, respectively (OR, 0.94; 95% CI, 0.50-1.79; p = .870). The miscarriage rate was greater in the isthmocele group (8%) than in the control group (4%); however, it did not reach statistical significance (OR, 2.09; 95% CI, 0.50-8.67; p = .302). The multiple pregnancy rate was similar between the groups (8% vs 5.3%, respectively; OR, 1.54; 95% CI, 0.41-5.70; p = .513). The groups were also similar in ectopic pregnancy rates (p = .560). These outcomes remained similar when adjusted for potential confounders on the regression analysis. CONCLUSION: Isthmocele does not seem to have a significant impact on the chance of pregnancy in assisted reproductive treatment settings. However, the embryo transfer procedure may be more difficult in the presence of an isthmocele.

Elevation of progesterone on the trigger day exerts no carryover effect on live birth in freeze-all cycles.

AIM: To evaluate the effect of trigger day progesterone (P) levels on live birth in freeze-all cycles. MATERIAL AND METHODS: Retrospective analysis of 1034 freeze-all female patients aged <38 years with single blastocyst transfers. Patients with (n = 268) or without (n = 766) preimplantation genetic test for aneuploidy (PGT-A) arm were further categorized into three subgroups based on trigger day P levels; low (<0.80 ng/ml), medium (0.8-1.49 ng/ml), and high (≥1.50 ng/ml). RESULTS: Estradiol (E2) levels on trigger day, the number of oocytes retrieved and the number of mature oocytes increased significantly with increasing serum p values in cycles without and with PGT-A arms. Significant correlation was found between E2 levels on trigger day and serum P levels and between the number of total oocytes retrieved and serum P levels Live birth rates were similar in the three subgroups in without PGT-A arm (51%, 52.6%, and 51.5%, respectively; p = .922) and with PGT-A arm (55.1%, 55.1%, and 62.5%, respectively; p = .730). Multivariate regression analysis revealed that trigger day P levels were not significant for live birth. CONCLUSION: The proposal that trigger day progesterone elevation (PE) exerts a detrimental effect on oocyte and embryo competence has no clinical validity.

Comparison of intramuscular versus subcutaneous aqueous progesterone for luteal phase support in artificially prepared frozen embryo transfer cycles.

OBJECTIVE: Cryopreservation of embryos for future transfer attempts has noticeably increased in the last decade, especially due to the technological developments in in vitro fertlization (IVF) laboratories. In parallel, different progesterone (P) replacement regimens preceding artificially prepared frozen embryo transfer (AC-FET) attempts, especially with respect to the route of application and dosing scheme, have been widely argued so far. We aimed to provide more information about the efficacy profile of novel subcutaneous aqueous progesterone (SP) in AC-FET cycles. MATERIALS AND METHODS: This retrospective, single-centre cohort study included a total of 507 AC-FET cycles performed between June 2018 and April 2020. Three hundred forty-nine (68.8%) patients received 50 mg of intramuscular progesterone as once daily, 158 (31.2%) patients received 25 mg of SP as twice daily. Only, the first and single blastocyst transfers from the same cohort were accepted. The inclusion criteria were as follows: females aged <37 years, body mass index ≥18 kg/m2 and ≤35 kg/m2, sperm concentration ≥5x106/mL. Pre-implantation genetic testing cycles were not included. The primary outcome was the live birth rate (LBR). RESULTS: The number of previous IVF attempts, type of infertility, peak estradiol (E2) levels, the total number of retrieved oocytes, mature oocytes, and the number of 2PN was significantly different between the groups. Positive pregnancy (p=0.474) and clinical pregnancy rates (p=0.979), LBR (p=0.404), and missed abortion rates (p=0.144) were comparable between the groups. The total number of oocytes [adjusted odds ratios (AOR)=1.024, 95% confidence interval (CI): 1.002-1.047; p=0.03)], endometrial thickness (AOR=1.121, 95% CI: 1.003-1.253; p=0.044), and cryopreservation day 5/6 (AOR=0.421, 95% CI: 0.226-0.788; p=0.007) achieved statistical significance following binary logistic regression analysis. However, P administration type did not achieve statistical significance (p=0.731). CONCLUSION: As a novel option, SP has comparable efficacy in pregnancy outcomes and may be accepted as an alternative for luteal phase support in AC-FET cycles.

Preimplantation genetic testing for aneuploidy in severe male factor infertility.

RESEARCH QUESTION: Does the use of preimplantation genetic testing for aneuploidies (PGT-A) improve outcomes in couples with severe male factor infertility (SMFI)? DESIGN: This retrospective cohort study included SMFI cases that underwent blastocyst transfer with/without PGT-A. Inclusion criteria were SMFI (azoospermia and sperm count <1 million/ml), women aged 25-39 years, single vitrified-warmed blastocyst transfer, and no intracavitary pathologies. Patients were divided into PGT-A and non-PGT-A groups. The primary outcome was live birth rate (live birth of an infant after 24 weeks of gestation); secondary outcomes were implantation and clinical pregnancy rates. RESULTS: The study included 266 SMFI cases (90 and 176 in the PGT-A and non-PGT-A groups, respectively). Men and women in the PGT-A group were significantly older than those in the non-PGT-A group. The groups did not differ in terms of male factor categories, sperm collection methods or additional female factors. Live birth rates in the PGT-A and non-PGT-A groups were 55.6% and 51.1%, respectively (odds ratio [OR] 1.19, 95% confidence interval [CI] 0.71-1.98, P = 0.495). The implantation rates were 65.6% and 64.2%, respectively (OR 1.06, 95% CI 0.62-1.80, P = 0.827). The clinical pregnancy rates were 62.2% and 58.0%, respectively (OR 1.19, 95% CI 0.71-2.01, P = 0.502). The use of PGT-A was not an independent factor for live birth (aOR 1.33, 95% CI 0.66-2.70, P = 0.421). Advanced age in women was the only independent factor associated with live birth (aOR 0.46, 95% CI 0.22-0.96, P = 0.041). CONCLUSIONS: The use of PGT-A does not seem to be an independent factor associated with live birth per transfer in couples with SMFI.

A 5-year multicentre randomized controlled trial comparing personalized, frozen and fresh blastocyst transfer in IVF.

RESEARCH QUESTION: Does clinical performance of personalized embryo transfer (PET) guided by endometrial receptivity analysis (ERA) differ from frozen embryo transfer (FET) or fresh embryo transfer in infertile patients undergoing IVF? DESIGN: Multicentre, open-label randomized controlled trial; 458 patients aged 37 years or younger undergoing IVF with blastocyst transfer at first appointment were randomized to PET guided by ERA, FET or fresh embryo transfer in 16 reproductive clinics. RESULTS: Clinical outcomes by intention-to-treat analysis were comparable, but cumulative pregnancy rate was significantly higher in the PET (93.6%) compared with FET (79.7%) (P = 0.0005) and fresh embryo transfer groups (80.7%) (P = 0.0013). Analysis per protocol demonstrates that live birth rates at first embryo transfer were 56.2% in PET versus 42.4% in FET (P = 0.09), and 45.7% in fresh embryo transfer groups (P = 0.17). Cumulative live birth rates after 12 months were 71.2% in PET versus 55.4% in FET (P = 0.04), and 48.9% in fresh embryo transfer (P = 0.003). Pregnancy rates at the first embryo transfer in PET, FET and fresh embryo transfer arms were 72.5% versus 54.3% (P = 0.01) and 58.5% (P = 0.05), respectively. Implantation rates at first embryo transfer were 57.3% versus 43.2% (P = 0.03), and 38.6% (P = 0.004), respectively. Obstetrical outcomes, type of delivery and neonatal outcomes were similar in all groups. CONCLUSIONS: Despite 50% of patients dropping out compared with 30% initially planned, per protocol analysis demonstrates statistically significant improvement in pregnancy, implantation and cumulative live birth rates in PET compared with FET and fresh embryo transfer arms, indicating the potential utility of PET guided by the ERA test at the first appointment.

Impact of elective frozen vs. fresh embryo transfer strategies on cumulative live birth: Do deleterious effects still exist in normal & hyper responders?

BACKGROUND: Is freeze-all strategy effective in terms of cumulative live birth rates (CLBRs) in all patients? METHODS: This retrospective single-center study analyzed the CLBRs of 2523 patients undergoing fresh or electively frozen blastocyst transfer cycles. In 1047, cycles, the fresh embryo transfer (ET) strategy was applied for the 1st ET, whereas electively frozen ET (e-FET) was performed in 1476 cycles. Female age ≤ 37 and blastocysts frozen via vitrification were included. The patients in each arm were further stratified into four subgroups according to the number of oocytes retrieved as follows: Group A: 1-5, group B: 6-10, group C: 11-15 and group D: 16-25 oocytes retrieved. The primary endpoint was the CLBR. The secondary endpoints were the ovarian hyperstimulation syndrome (OHSS) rate and the live birth rates (LBRs) following fresh ETs and e-FETs for the first transfers. RESULT(S): The CLBR was similar between the fresh ET and e-FET arms in group A (35/76 (46.1%) vs 29/67 (43.3%), p = 0.74) and group B (165/275 (60%) vs 216/324 (66.7%), p = 0.091), whereas significantly higher rates were detected in favor of the e-FET arm within group C (328/460 (71.3%) vs 201/348 (57.8%), p<0.001) and group D (227/348 (65.2%), vs 446/625 (71.5%), p<0.001). The OHSS rate was also found to be higher in the fresh ET arm among group C (12/348 (3.4%) vs 0/460 (0%), p<0.001) and group D (38/348 (10.9%) vs 3/625 (0.5%), p<0.001) patients than e-FET arm. Perinatal and obstetrical outcomes were nonsignificantly different between fresh and e-FET arms. However, the birth weights were significantly lower for fresh ET, 3064 versus 3201 g for singletons (p<0.001). CONCLUSION: Compared with a fresh-transfer strategy, the e-FET strategy resulted in a higher CLBR among patients with >10 oocytes retrieved during stimulated cycles.