Reproductive Center Research Articles

P-282 Decreased 14,15-EET in follicle induced ovary granulosa cells senescence and contributes to endometriosis-associated infertility

Abstract Study question What is the expression profile of lipid peroxides in endometriosis (EM) follicle? What is the function of differentially expressed lipid peroxides in endometriosis-associated infertility? Summary answer 14,15-EET concentration is decreased in EM granulosa cells (GCs) and follicular fluid (FF), decreased 14,15-EET in EM induced GCs senescence by activating PI3K-AKT-mTOR signaling pathway. What is known already Oxidative stress (OS) induced EM GCs senescence and intensified peroxidation in the follicle compromise the development of oocyte. Lipid peroxidation products generated in follicle cyclically and can be used as OS markers to evaluate Assisted Reproductive Technology (ART) outcomes. Epoxyeicosatrienoic acids (EETs) is one of the mostly studied lipid peroxides that derived from arachidonic acid (AA) and metabolized to less potent DHETs by soluble epoxide hydrolase (EPHX2). 14,15-EET was considered to be a active molecule that exert anti-reactive oxygen species, anti-inflammatory, anti-migratory, and pro-fibrinolytic effects in cardiovascular disease, kidney injury and metabolic syndrome. But no reports were found in endometriosis. Study design, size, duration This study was started on January 1, 2022. We enrolled 101 infertility patients who underwent IVF-ET at our reproductive center to explore the profile of lipid peroxides in follicle. The mechanism of decreased EETs in endometriosis follicle was studied on human GCs and mouse GCs via quantitative reverse transcriptase PCR, western blot, enzyme-linked immunosorbent assay, liquid chromatography-tandem mass spectrometry (LC-MS/MS). Endometriosis mouse model and mouse GCs were used to verify the anti-senescence role of 14,15-EET. Participants/materials, setting, methods 141 lipid peroxides were measured in FF from 56 EM patients and 45 infertility women with salpingectomy (Control) by LC-MS/MS. Linear regression analysis was conduct to explore the correlations between EETs and ART outcomes. TPPU, an EPHX2 enzyme inhibitor, was used in vitro and in vivo to explore the function of 14, 15-EET in GCs. RNA-Sequencing was performed to find the mechanism by which reduced 14,15-EET leads to abnormal folliculogenesis. Main results and the role of chance LC-MS/MS results showed that 15 lipid peroxides differentially expressed in EM-FF, and all of them were downregulated in EM-FF when compared with control FF. The levels of 4 different regioisomers of EETs (5,6-EET, 8,9-EET, 11,12-EET and 14,15-EET) in EM patients were significant lower than that in control women (P < 0.05). Linear regression analysis showed positive correlations between EETs scores and ART outcomes. Decreased EETs in endometriosis follicular fluid indicates lower numbers of retrieved oocytes, mature oocytes, total embryos and high-quality embryos. 14,15-EET addition alleviated OS-induced GCs senescence as well as mitochondrial membrane potential (MMP) and adenosine triphosphate (ATP) reduction in GCs. Moreover, TPPU administration lead to increased 14,15-EET expression and rescued cellular senescence, MMP and ATP abnormities of endometriosis GCs in vitro and in vivo. RNA-Sequencing results revealed significantly suppression of PI3K-AKT-mTOR signaling pathway in GCs after TPPU administration. Western blot results indicated that 14,15-EET alleviate GCs senescence via inhibiting PI3K-AKT-mTOR signaling pathway. Limitations, reasons for caution 14,15-EET is just one of the lipid peroxides in FF and GCs. A variety of lipid peroxides expressed in follicle, and more research were needed to explore the roles of them. Wider implications of the findings This research makes the supplement of EETs during IVF-ET become meaningful and TPPU may represent a novel adjuvant therapy strategy for endometriosis-associated infertility. Trial registration number Not applied

P-668 Progesterone levels on the thawed embryo transfer day in artificial cycles with vaginal progesterone are not a predictor of clinical pregnancy: a prospective study

Abstract Study question Are serum progesterone (P) levels on the thawed embryo transfer day a good predictor of clinical pregnancy with own oocytes in artificial endometrial preparation cycles? Summary answer In artificial endometrial preparation cycles, serum P levels on the thawed embryo transfer day are not a good predictor of clinical pregnancy with own oocytes. What is known already Some studies have shown that low serum P levels on the thawed embryo transfer (ET) day are associated with a lower pregnancy rate after artificial endometrial preparation (AEP) cycles. Other studies have shown that serum P is not a good predictor of pregnancy rates. Furthermore, studies are inconclusive about which cutoff level for P on ET day correlates with better gestational outcomes. Study design, size, duration Prospective cohort study with 235 patients who underwent ET after AEP cycle with estradiol valerate and vaginal micronized P (200 mg/ 8 h). All patients underwent blastocyst-stage or cleavage-stage ET with embryos from their own oocytes, without preimplantation genetic testing for aneuploidies, after AEP cycle, with a triple layer endometrium > 7 mm. The study was carried out at the Human Reproduction Center of Ribeirão Preto Clinical Hospital from November 2021 to June 2023. Participants/materials, setting, methods Blood was collected between 8–10 am on the thawed ET day and serum P concentration was measured. The primary outcome was clinical pregnancy (CP) beyond pregnancy week 8. A ROC curve detected the best P cutoff level that correlated with higher CP rate. Thereafter, patients were categorized into those with serum P above and below the cutoff level. Multivariate logistic regression was performed to verify which variables were predictors of CP and P levels. Main results and the role of chance Serum P measured on the ET day showed no significant difference between patients who had CP and those who did not (12.76 +4.1 versus 11.9 + 5.6, p 0.3318). The cutoff level for serum P measured on the ET day above which correlates with higher CP rate was 10.6 ng/mL (AUC 0.58), with a sensibility of 72% and specificity of 44%. Patients who had serum P on the day of ET above this value had a significantly higher CP rate than those who had it below this value (25.9% versus 14.6%) (p 0.0372). However, when the multivariate logistic regression model was performed, there was no significant difference in the CP between the groups (OR 2, 95%IC: 0.9-5). Predictors of CP in the study were the presence of at least one top-quality embryo transferred (OR 2.6, 95%IC: 1-6.6) and the transfer of embryo at the blastocyst-stage (OR 5.2, 95%IC: 1.9-14). The presence of endometriosis was negatively associated with CP (OR 0.2, 95%IC: 0.09-0.9). Limitations, reasons for caution The small sample size is a limitation. We included the blastocyst-stage and the cleavage-stage embryo transfer. Only women with vaginal micronized P were included. Extrapolation to other P administration forms needs to be evaluated. Wider implications of the findings Monitoring serum P levels on the day of ET in AEP cycles with vaginal micronized P is not justified as serum P levels are not a good predictor of CP in cycles with own oocytes. Trial registration number Not applicable

Vitamin D levels in couples undergoing in vitro fertilization treatment: lack of association with embryo quality or pregnancy rates

ObjectiveTo investigate the potential impact of vitamin D serum levels of couples going through in vitro fertilization in terms of embryo quality and pregnancy rates. DesignRetrospective cohort study SettingFertipraxis, private human reproduction center on Rio de Janeiro, Brazil. Subjects267 couples who underwent intracytoplasmic sperm injection between January 2017 and March 2019. ExposureThe couples were categorized into four groups based on 25OH vitamin D levels measured at the beginning of the stimulation protocol: Group 1 with levels ≥ 30 ng/mL for both women and men; Group 2 with levels < 30 ng/mL for both; Group 3 with women < 30 ng/mL and men ≥ 30 ng/mL; and Group 4 with women ≥ 30 ng/mL and men < 30 ng/mL. Main Outcome MeasuresWe consider quantity and quality of embryos during the cleavage and blastocyst stages as primary outcomes. Correspondingly, clinical pregnancy rate was regarded as a secondary outcome. ResultsOur findings revealed no significant correlations between the studied VD groups and the evaluated outcomes. This includes quantity and quality of embryos during the cleavage and blastocyst stages, as well as clinical pregnancy rate. Primary analysis revealed a small but statistically significant difference in the duration of controlled ovarian stimulation between group 1 and group 2 (p=0.035; CI=0.07 – 3.04) and between group 1 and group 3 (p=0.040; CI=0.05 – 3.23). ConclusionThe present study found no correlation between the studied VD levels and quantity and quality of cleavage or blastocyst stage embryos, nor did it show any impact on clinical pregnancy rates. Further well designed, prospective studies are warranted to determine whether and how vitamin D affects reproductive outcomes.

Маркування ведмедем (Ursus arctos) дерев у Чорнобильському біосферному заповіднику

For the first time in the Ukrainian Polissia and the Chornobyl Radiation and Ecological Biosphere Reserve (ChREBR), a concentrated linear distribution of bear-marked spruce trees was found at the abandoned estates of the village of Opachychi along the second terrace of the Prypiat River floodplain. Individual sightings of bears (Ursus arctos) in the Ukrainian Polissia have been known since the 1990s, but there are no reliable data on the reproduction of this species. The abundance of bears in Belarus has increased in recent years, which leads to the possibility of females coming to the Polissia from the north. Dried spruce trees without resin were not marked by the bear, and the animal moved on to live trees. Friction and more intensive marking of trees was recorded mainly in the eastern direction. Most of the marked spruce trees were upright with no branches to the height of the marking. The intensive marking behaviour of bears in 2023 in the ChREBR can be considered as the appearance of a female, as signalling to a female, and as a sign of dominance towards a male competitor. Two adult males were observed near one of the marked trees (data from amateurs). Special field surveys are needed to identify females with cubs to confirm or refute the hypothesis that the intensity of bear marking can be used in the event of the presence of a female bear in oestrus. Along the bear-marked trees in the village of Opachychi of the ChPEBR, an ecological trail and a visitor centre of large carnivores and herbivores should be created, which would be popular destinations. It is advisable to arrange a location for wildlife observations in an abandoned house for photo and video shooting, as is usually done in Sweden, Bulgaria, Romania, and Spain. In such tours, the effectiveness of the observation is not guaranteed and only preliminary statistics of wildlife encounters are provided. For this purpose, it is advisable to create two feeding areas for predatory mammals with dry food, as well as a salt marsh and a feeding trough for ungulates. The majority of the ChREBR territory has a low incidence of markings. The bear in the Polissia, as elsewhere in Europe, is expanding its range naturally, but at a slow pace due to the low intensity of female dispersal. Tree marking by bears was also observed in Chernihiv Oblast, in the Mizhrichynskyi Regional Landscape Park. It is advisable to release several bears in the ChEBR to create a reproductive centre for this species in the Polissia.

Effect of body mass index on progesterone level on trigger day in gonadotropin-releasing hormone antagonist cycles

Objective To investigate the effect of body mass index (BMI) on progesterone (P) level on trigger day in gonadotropin-releasing hormone antagonist (GnRH-ant) cycles. Methods This study was a retrospective cohort study. From October 2017 to April 2022, 412 in-vitro fertilization (IVF)/intracytoplasmic sperm injection (ICSI) patients who were treated with GnRH-ant protocol for controlled ovarian hyperstimulation (COH) in the reproductive center of our hospital were selected as the research objects. Patients were divided into three groups according to BMI level: normal weight group (n = 230):18.5 kg/m2≤BMI < 24 kg/m2; overweight group (n = 122): 24 kg/m2≤BMI < 28 kg/m2; Obesity group (n = 60): BMI ≥ 28 kg/m2. Variables with p < .10 in univariate analysis (BMI, basal FSH, basal P, FSH days, Gn starting dose and E2 level on trigger day) and variables that may affect P level on trigger day (infertility factors, basal LH, total FSH, HMG days and total HMG) were included in the multivariate logistic regression model to analyze the effect of BMI on P level on trigger day of GnRH-ant protocol. Results After adjustment for confounding factors, compared with that in normal weight patients, the risk of serum P elevation on trigger day was significantly lower in overweight and obese patients (OR = 0.434 and 0.199, respectively, p < .05). Conclusion The risk of P elevation on trigger day in GnRH-ant cycles decreased with the increase of BMI, and BMI could be used as one of the predictors of P level on trigger day in GnRH-ant cycles.

Differential expression of CYP19A1 and lncRNA-CTBP1-AS in the granulosa cells of women with polycystic ovary syndrome

Background: Hyperandrogenism is the basic indicator of polycystic ovarian syndrome (PCOS) and a key factor in its pathological processes. Patients with PCOS have higher testosterone levels along with a decline in the estradiol/testosterone ratio, which may indicate an aromatase dysfunction. The aim of the study: Investigating the expression levels of CYP19A1 and lncRNA-CTBP1-AS in the granulosa cells (GCs) of women with and without PCOS. Materials and methods: We compared the expression levels of CYP19A1 and lncRNA-CTBP1-AS in granulosa cells from 16 PCOS patients and 20 controls who underwent assisted reproductive technology (ART) treatment at the Center for Human Reproduction and IVF (Rostov-on-Don, Russia) using quantitative real-time PCR. Results: We observed a significant downregulation of CYP19A1 expression in the GCs of PCOS patients, with expression levels approximately 4.7 times lower compared to the control group. Furthermore, we found a positive correlation between CYP19A1 expression and the E2/T ratio in PCOS patients. Conversely, PCOS patients exhibited a remarkable upregulation of CTBP1-AS expression in their GCs, with levels up to 23 times higher than those in the control group. Importantly, we also identified a significant negative correlation between the expression level of CTBP1-AS and CYP19A1 in the GCs of PCOS patients. Conclusion: Our results indicate a potential regulatory role of CTBP1-AS in CYP19A1 expression and suggest its involvement in the pathogenesis of PCOS.

Антиоксидантная терапия в коррекции ДНК-фрагментации сперматозоидов.

Introduction. The progressive deterioration of reproductive function, observed throughout the world in recent decades, is an extremely alarming trend, while the incidence of male infertility is increasing. The universal pathogenetic mechanism of spermatogenesis disorders, which underlies almost all forms of male infertility, is oxidative stress of sperm. It has various manifestations, the essence of which, as a rule, boils down to the formation of mitochondrial dysfunction and fragmentation of sperm DNA. It has been found that consuming antioxidants and microelements helps increase the chances of conception for subfertile couples, and also reduces the risk of reproductive losses. Of greatest and justifiable interest are medicinal complexes designed to reduce the level of oxidative stress and sperm DNA fragmentation. The purpose of the study was to evaluate the effect of antioxidant therapy on the degree of sperm DNA fragmentation in men suffering from infertility. Materials and methods. The study involved 42 men aged 25 to 37 years with an established diagnosis of male infertility who were treated at the Center for Family Health and Reproduction. The patients were divided into 2 groups, comparable according to the main criteria assessed. Patients in the control group (12 people, 29%) received standard antioxidant therapy. Patients of the main group (30 people, 71%) received the BESTFertil-DHA® complex. The course of treatment for patients in both groups was 3 months. Control examinations were carried out at 1.5 and 3 months from the start of therapy. Spermogram data and assessment of the degree of DNA fragmentation in sperm using the SCD (Sperm chromatin dispersion test) method were used as the main criterion for assessing the effectiveness of therapy. Results. After three months of taking BESTFertil-DHA®, in 16 patients (53%), the degree of sperm DNA fragmentation decreased to less than 15%. Against this background, 5 couples became pregnant naturally, and 4 couples became pregnant as a result of the in vitro fertilization procedure. The average level of sperm DNA fragmentation was less than 13% (p &lt; 0.05). A decrease in the level of DNA fragmentation was noted in 53% of patients after 1.5 months of taking BESTFertil-DHA®, which is especially important when preparing patients for assisted reproductive technology programs. In patients taking BESTFertil-DHA®, the total progressive motility of spermatozoa «A+B» increased by 28%, and the concentration of spermatozoa in 1 ml of ejaculate – by 61%. In the control group, the degree of sperm DNA fragmentation did not undergo statistically significant changes during treatment. Conclusion. BESTFertil-DHA® statistically significantly helps to reduce the degree of DNA fragmentation in sperm, increase the number of actively motile forms of sperm and reduce the degree of teratozoospermia.

Nomogram to predict the probability of clinical pregnancy in women with poor ovarian response undergoing in vitro fertilization/ intracytoplasmic sperm injection cycles.

Poor ovarian response (POR) is associated with decreased clinical pregnancy rates, emphasizing the need for developing clinical prediction models. Such models can improve prognostic accuracy, personalize medical interventions, and ultimately enhance live birth rates among patients with POR. This study aims to develop and validate a prognostic model for predicting clinical pregnancy outcomes in individuals with POR undergoing in vitro fertilization/ intracytoplasmic sperm injection (IVF/ICSI) cycles. A retrospective cohort of 969 patients with POR undergoing fresh embryo transfer cycles at the Reproductive Center of Fujian Maternal and Child Health Center from January 2018 to January 2022 was included. The cohort was randomly divided into model (n = 678) and validation (n = 291) groups in a 7:3 ratio. A single-factor analysis was performed on the model group to identify variables influencing clinical pregnancy. Optimal variables were selected using LASSO regression, and a clinical prediction model was constructed using multivariate logistic regression analysis. The model's calibration and discrimination were assessed using receiver operating characteristic (ROC) and calibration curves, while the clinical utility was evaluated using decision curve analysis. Multivariate logistic regression analysis revealed that the age of the women (odds ratio [OR] 0.936, 95% confidence interval [CI] 0.898-0.976, P = 0.002), body mass index (BMI) ≤ 24 (OR 2.748, 95% CI 1.724-4.492, P < 0.001), antral follicle count (AFC) (OR 1.232, 95% CI 1.073-1.416, P = 0.003), anti-Müllerian hormone (AMH) (OR 1.67, 95% CI 1.178-2.376, P = 0.004), number of mature oocytes (OR 1.227, 95% CI 1.075-1.403, P = 0.003), number of embryos transferred (OR 1.692, 95% CI 1.132-2.545, P = 0.011), and transfer of high-quality embryos (OR 3.452, 95% CI 1.548-8.842, P = 0.005) were independent predictors of clinical pregnancy in patients with POR. According to the receiver operating characteristic (ROC) analysis, the prediction model exhibited an area under the curve (AUC) of 0.752 (0.714, 0.789) in the model group and 0.765 (0.708, 0.821) in the validation group. The clinical decision curve demonstrated that the model held maximum clinical utility in both cohorts when the threshold probability of clinical pregnancy ranged from 6-81% to 12-82%, respectively. Clinical pregnancy outcomes in patients with POR who underwent IVF/ICSI treatment were influenced by several independent factors, including the age of the women, BMI, AFC, AMH, number of mature oocytes, number of embryos transferred, and transfer of high-quality embryos. A clinical prediction model based on these factors exhibited favorable clinical predictive and applicative value. Therefore, this model can serve as a valuable tool for clinical prognosis, intervention, and facilitating personalized medical treatment.

Clinical factors impacting microdissection testicular sperm extraction success in hypogonadal men with nonobstructive azoospermia

To explore factors influencing microdissection testicular sperm extraction (micro-TESE) success in hypogonadal men with nonobstructive azoospermia (NOA). A cohort study. University-affiliated male reproductive health center. A total of 616 consecutive patients with NOA and hypogonadism (total testosterone [T] levels <350 ng/dL) underwent micro-TESE between 2014 and 2021. All patients had no prior sperm retrieval (SR) history. Patients aged 23-55 years underwent comprehensive clinical, laboratory, and histopathological diagnostic evaluation for NOA and were further categorized into two cohorts on the basis of pre-SR hormonal stimulation. A multivariable logistic regression analysis explored the associations between patient variables and micro-TESE success, defined as the presence of viable spermatozoa in extracted specimens. Adjusted odds ratios (aORs) and 95% confidence intervals (CIs) were computed to assess the relationship between SR success and relevant predictors. Sperm retrieval rates were compared between patients receiving or not hormonal stimulation, and logistic regression analysis evaluated the effect of baseline follicle-stimulating hormone levels (i.e., normogonadotropic vs. hypergonadotropic classes) on SR success. The overall micro-TESE success rate was 56.6%. Baseline follicle-stimulating hormone levels (aOR, 0.97; 95% CI, 0.94-0.99), pre-SR hormonal stimulation (aOR, 2.54; 95% CI, 1.64-3.93), presence of clinical varicocele (aOR, 0.05; 95% CI, 0.01-0.51), history of previous varicocelectomy (aOR, 2.55; 95% CI, 1.26-5.16), and testicular histopathology were independent predictors of SR success. Among hormone-pretreated patients, pre-micro-TESE T levels and delta T (an absolute increase in T levels from baseline) were associated with SR success. A pre-micro-TESE T level of 418.5 ng/dL (area under the curve value: 0.78) and a delta T of 258 ng/dL (area under the value: 0.76) distinguished patients with positive and negative SR outcomes. Subgroup analysis showed that pre-SR hormonal stimulation yielded a greater benefit for normogonadotropic patients than for those who were hypergonadotropic. This study underscores the association between clinical factors and micro-TESE success in hypogonadal men with NOA. Although causality is not established, our findings suggest that these patients may benefit from pre-SR interventions, particularly hormonal stimulation and varicocele repair. NCT05110391.

Complementary and Alternative Treatment Methods Used by Infertile Women in Turkiye: A Cross-Sectional Study

Objective: This study aims to determine the complementary and alternative therapy (CAM) methods used among infertile women in Turkiye, the prevalence of their use and the factors affecting their use. Method: The sample of the descriptive and cross-sectional study consisted of 142 women who applied to the Assisted Reproductive Treatment Centre of a public hospital and volunteered to participate in the study. The data were collected with "Personal Information Form" and "Complementary and Alternative Medicine Approaches Scale (CAMAS)". Descriptive statistics, Mann-Whitney U, Kruskal Wallis Test were used in the evaluation of the data. Results: While the average age of the women who contributed to the study was found to be 29.77±4.83 years. When the CAM methods used by women are examined, they are mostly prayer (88.0%), prayer (namaz) (78.2%), vow (25.4%), visit to the tomb (24.6%), amulet (20.4%), black pepper (17.6%), ginseng (15.5%), astralagus (13.4%), going to a teacher and having yourself read (13.4%), shark cartilage (13.4%), turtle blood (13.4%), lead casting (4.2%), reiki (2.8%), bioenergy (2.1%) and hypnosis it was determined to be (1.4%). When the mean scores in the CAMAS subscales were examined, it was determined that the highest score was body-mind approaches (40.46±17.37). Conclusion: In line with the research findings, it was determined that the use of CAM was common among infertile women in Turkiye and the most frequently used approaches were spiritual and herbal methods. Health professionals need to evaluate infertile women about their use of CAM methods and inform women about these methods.

The impact of sociocultural and psychological stress on the outcome of assisted reproductive technology in remarried families

Objective In China, there is a unique type of second marriage (SM) family where the woman is remarried, and the man is experiencing his first marriage. Additionally, the woman is older than the man. Therefore, these families experience many challenges: psychological, emotional, and societal pressure. Such family is a typical sample for studying sociocultural and psychological stress influencing on outcome of assisted reproductive technology (ART). This study aimed to investigate the impact of social psychological stress on the live birth outcomes AR. Methods In this retrospective cohort, second marriage (SM) families who visited the Second Hospital of Hebei Medical University reproductive center between January 2012 to December 2022 were screened, and 561s marriage families (the SM group) with 5600 first marriage (FM) families (the FM group) were included undergoing their first ART cycles. The primary outcome of this study was the live birth rate (LBR). Results The live birth rate (LBR) of SM group (30.7%) is lower than that of the FM group (43.6%) (p < 0.01). After adjustment by logistic regression, the second marriage group (OR = 1.269, 95%CI 1.031–1.562, p = 0.025) were independent factors associated with the outcome of live birth. After propensity score matching (PSM), the live birth rate of SM group (28.7%) is lower than the FM group (35.9%) (0 = 0.011). Conclusion The SM family experience higher levels of social and psychological pressure, which lead to lower level of LBR than FM family.

The Impact of Embryo Quality on Pregnancy Outcomes in Single Day 5 versus Day 6 Euploid Blastocyst Transfer: A Retrospective Cohort Study.

Selecting embryos with the highest implantation potential is crucial for in vitro fertilization (IVF) success. Both the timing of blastulation, day 5 (D5) or D6, and the embryo quality have been suggested as influential factors in determining the clinical outcome of single euploid blastocyst transfers. However, evidence supporting the superiority of D5 over D6 blastocysts remains inconclusive. The aim of this study was to compare clinical outcomes following the transfer of euploid blastocysts with different quality and timing of blastulation. A retrospective cohort study was conducted at our Assisted Reproductive Center, analyzing the outcome of 774 transfers with D5 euploids and 155 transfers with D6 euploids performed between January 2019 and February 2022. The live birth rate was significantly lower in the euploid D6 group compared to the euploid D5 group (38.71vs. 55.04%, P=0.001). The outcome was significantly influenced by the quality of the embryos. Live birth rates were 62.14 and 53.61% following transfers of D5 and D6 excellent embryos respectively, 45.18 and 32.21% following transfer of D5 and D6 good embryos but only 28.64 and 19.32% following transfer of D5 and D6 fair embryos. The outcome difference was statistically significant across embryo quality categories (P=0.001). The adjusted risk ratios (RR) of clinical outcomes indicated that excellent euploid D5 embryos consistently outperformed other types of embryo quality. The timing of blastulation and embryo quality are crucial factors in determining the success of single euploid blastocyst transfers. Excellent euploid D5 transfers yielded superior clinical outcomes, providing valuable insights for IVF teams and patients when selecting embryos to be transferred.

Progesterone levels do not differ between patients with or without endometriosis/adenomyosis both in those who conceive after hormone replacement therapy-frozen embryo transfer cycles and those who do not.

Do women with endometriosis who achieve a live birth (LB) after HRT-frozen embryo transfer (HRT-FET) have different progesterone levels on the day of transfer compared to unaffected women? In women achieving a LB after HRT-FET, serum progesterone levels on the day of the transfer did not differ between patients with endometriosis and unaffected patients. In HRT-FET, several studies have highlighted the correlation between serum progesterone levels at the time of FET and LB rates. In the pathophysiology of endometriosis, progesterone resistance is typically described in the eutopic endometrium. This has led to the hypothesis that women with endometriosis may require higher progesterone levels to achieve a LB, especially in HRT-FET cycles without a corpus luteum. We conducted an observational cohort study at the university-based reproductive medicine center of our institution, focusing on women who underwent a single autologous frozen blastocyst transfer after HRT using exogenous estradiol and micronized vaginal progesterone for endometrial preparation between January 2019 and December 2021. Women were included only once during the study period. Serum progesterone levels were measured on the morning of the FET by a single laboratory. Patients were divided into groups based on whether they had endometriosis or not and whether they achieved a LB. The diagnosis of endometriosis was based on published imaging criteria (transvaginal sonography/magnetic resonance imaging) and/or confirmed histology. The primary outcome was progesterone levels on the day of the HRT-FET leading to a LB in patients with endometriosis compared to unaffected women. Subgroup analyses were performed based on the presence of deep infiltrating endometriosis or adenomyosis. A total of 1784 patients were included. The mean age of the women was 35.1 ± 4.1 (SD) years. Five hundred and sixty women had endometriosis, while 1224 did not. About 179/560 (32.0%) with endometriosis and 381/1224 (31.2%) without endometriosis achieved a LB. Among women who achieved a LB after HRT-FET, there was no significant difference in the mean progesterone level on the day of the HRT-FET between those with endometriosis and those without (13.6 ± 4.3 ng/ml vs 13.2 ± 4.4 ng/ml, respectively; P = 0.302). In the subgroup of women with deep infiltrating endometriosis (n = 142) and adenomyosis (n = 100), the mean progesterone level was 13.1 ± 4.1 ng/ml and 12.6 ± 3.7 ng/ml, respectively, with no significant difference compared to endometriosis-free patients. After adjusting for BMI, parity, duration of infertility, tobacco use, and geographic origin, neither the presence of endometriosis (coefficient 0.38; 95% CI: -0.63 to 1.40; P = 0.457) nor the presence of adenomyosis (coefficient 0.97; 95% CI: -0.24 to 2.19; P = 0.114) was associated with the progesterone level on the day of HRT-FET. Among women who did not conceive, there was no significant difference in the mean progesterone level on the day of the HRT-FET between those with endometriosis and those without (P = 0.709). The primary limitation of our study is associated with its observational design. Extrapolating our results to other laboratories or different routes and/or dosages of administering progesterone also requires validation. This study shows that patients diagnosed with endometriosis do not require higher progesterone levels on the day of a frozen blastocyst transfer to achieve a LB in hormonal replacement therapy cycles. None declared. N/A.

The SCREENIVF Hungarian version is a valid and reliable measure accurately predicting possible depression in female infertility patients

Infertility patients, often in high distress, are entitled to being informed about their mental status compared to normative data. The objective of this study was to revalidate and test the accuracy of the SCREENIVF, a self-reported tool for screening psychological maladjustment in the assisted reproduction context. A cross-sectional, questionnaire-based online survey was carried out between December 2019 and February 2023 in a consecutive sample of female patients (N = 645, response rate 22.9%) in a university-based assisted reproduction center in Hungary. Confirmatory factor analysis and cluster and ROC analyses were applied to test validity, sensitivity and specificity in relation to Beck Depression Inventory (BDI) scores. Model fit was optimal (chi-square = 630.866, p < 0.001; comparative fit index = 0.99; root-mean-square error of approximation = 0.018 (90% CI 0.013–0.023); standardized-root-mean-square-residual = 0.044), and all dimensions were reliable (α > 0.80). A specific combination of cutoffs correctly predicted 87.4% of BDI-scores possibly indicative of moderate-to-severe depression (χ2(1) = 220.608, p < 0.001, Nagelkerke R2 = 0.462, J = 66.4). The Hungarian version of the SCREENIVF is a valid and reliable tool, with high accuracy in predicting BDI-scores. Low response rate may affect generalizability. The same instrument with different cutoffs can serve various clinical goals.

Gonadotropin-Releasing Hormone (GnRH) Agonist Protocol Improves Pregnancy Outcomes During In Vitro Fertilization (IVF) and Intracytoplasmic Sperm Injection (ICSI) Treatment in Young Infertile Women: A Retrospective Study.

Objective The objective of this study was to determine if gonadotropin-releasing hormoneagonist (GnRH-a) orgonadotropin-releasing hormoneantagonist (GnRH-ant) protocols during in vitrofertilization (IVF) or intracytoplasmic sperm injection (ICSI)treatment in younginfertile women improvetheir pregnancy outcomes. Methodology We retrospectively reviewed the records of 876young infertile women aged 20-35 years who underwentfresh embryo transfer in IVF/ICSI cycles.The data were collected from their initial visits to the reproductive medicine center ofthe Second Affiliated Hospital of Zhengzhou University between January 2019 and December 2022. We divided them into two groups according to the controlled ovarian hyperstimulation (COH) protocols: GnRH-a (n = 580) and GnRH-ant (n = 296).The primary outcome assessed in this study was the live birth rate. The secondary observation indicators included the total dose and duration of gonadotropin (Gn),total embryo transfer,day three (D3) embryo transfer, total two pro-nuclei (2PN) cleavage count, number of fertilizations, and implantation rate. Results The live birth rate had no clinical significance (P> 0.05). The total dose and duration of Gn stimulation in the GnRH-ant group were lower than in the GnRH-a group (P < 0.05). The total embryo transfer, D3 embryo transfer, total cleavage count, total 2PN cleavage count, number of fertilizations, transfer, and mature oocytes in metaphase II (MII) of D3 embryos in the GnRH-a group were higher than those in the GnRH-ant group (P < 0.05). The clinical pregnancy rate and implantation rate of the GnRH-a group were higher than those of the control group. Conclusions The total embryo transfer, D3 embryo transfer, total cleavage count, total 2PN cleavage count, number of fertilizations, transfer and MII of D3 embryos, clinical pregnancy, and implantation rateswere significantly higher in the GnRH-a protocol group.The total dosage of Gn and duration of Gn stimulation were lower in the GnRH-ant group than in the GnRH-a group. These findings provide the basis for the selection of the COH protocol in normal Chinese ovarian response patients undergoing IVF/ICSI.

Adherence to dietary recommendations mediates the effect of affective temperaments on infertility treatment outcomes

Affective temperaments have been shown to robustly affect infertility treatment success. However, identification of possible mediating factors through which they exert their influence is still lacking. A growing number of results suggest that adherence to recommended treatments may be such a mediator, on the one hand, because affective temperaments are known to influence adherence and, on the other hand, because non-adherence negatively influences the treatment outcome. Recommended treatment of infertility involves, beyond medications, dietary and lifestyle changes. The aim of this retrospective cohort study was to evaluate whether adherence to physician-prescribed diet and physical activity recommendations mediates the effect of affective temperaments on infertility treatment outcomes. Among 308 women who underwent infertility treatment in an Assisted Reproduction Center, affective temperaments, adherence to diet, adherence to physical exercise, and infertility treatment success (clinical pregnancy) were assessed besides detailed medical history and demographic parameters. Associations between affective temperaments, adherence to diet and recommended physical activity, and assisted reproduction outcomes were analyzed using generalized linear models and causal mediation analysis. Adherence to physical activity didn’t have an effect, but diet adherence increased the odds of infertility treatment success by 130% suggesting its role as a potential mediator. Based on causal mediation analysis, higher depressive and anxious temperament scores were directly associated with 63% and 45% lower odds of achieving clinical pregnancy, respectively, with effects not mediated by diet adherence. Higher irritable temperament scores indirectly decreased the odds of achieving clinical pregnancy by 14%, mediated by diet adherence; while higher cyclothymic temperament scores decreased the odds of achieving clinical pregnancy both directly by 51% and indirectly, mediated by diet adherence by 11%. Our results suggest that diet adherence mediates the mechanism by which irritable and cyclothymic affective temperaments influence IVF treatment success. Since adherence is a modifiable risk factor of infertility treatment success, screening for affective temperaments may help to identify potentially high-risk non-adherent patient groups and offer patient-tailored treatment, which may help increase the chances of a successful pregnancy and live birth in women undergoing IVF treatment.

Urinary haloacetic acid concentrations in relation to sex and thyroid hormones among reproductive-aged men

Sex and thyroid hormones are critical for male reproductive health. However, the associations between haloacetic acid (HAA) exposure – a known endocrine disruptor – and sex and thyroid hormones in humans remains unclear. We thus recruited 502 male participants seeking fertility evaluation from a reproductive center. We measured concentrations of sex and thyroid hormones in a single blood sample and dichloroacetic acid (DCAA) and trichloroacetic acid (TCAA) in repeated urine samples. Multivariable linear regression models were constructed to evaluate the associations between HAA concentrations and hormone measurements. After adjusting for potential confounders and urinary creatinine concentrations, urinary concentrations of TCAA were inversely associated with serum levels of sex hormone-binding globulin (SHBG), testosterone (T), T/luteinizing hormone ratio (T/LH), and thyroid stimulating hormone (TSH) (all P for trend < 0.10). Compared with participants in the lowest quartile of TCAA concentrations, those in the highest quartile had reduced serum levels of SHGB by 14.2 % (95% CI: −26.7, −3.0 %), T by 11.1 % (95% CI: −21.7, −1.3 %), T/LH by 21.0 % (95% CI: −36.7, −7.1 %), and TSH by 19.1 % (95% CI: −39.7, −1.5 %). Additionally, we observed inverse associations between continuous measurements of urinary HAAs and serum levels of free T, bioactive T, and estradiol. Our findings suggest that male HAA exposure may be associated with disrupted sex and thyroid function.

The risk of asthma in singletons conceived by ART: a retrospective cohort study.

Do singleton children conceived by ART have a higher asthma risk than naturally conceived (NC) singletons? The asthma risk was similar for ART-conceived singletons and NC singletons, and there were no clear differences between the various types of ART. Whether ART increases asthma risk in offspring is questionable. The evidence is inconsistent and limited by ethnicity, geographic distribution, inadequate confounder adjustment, unsatisfactory control groups, and specific methods of ART. Furthermore, the mediating effects of obstetric and neonatal outcomes on the association between ART and asthma remain unclear. This observational, single-centre study was conducted at a reproductive centre of an affiliated university hospital between September 2009 and April 2023. A total of 3227 singletons aged 3-6 years conceived by IVF versus ICSI or fresh versus frozen embryo transfer were retrospectively enrolled, and a total of 1206 NC singletons of the same age were subsequently recruited. Asthma was defined as a self-reported physician diagnosis or wheezing in the past 12 months. We performed multivariable logistic regression analyses to examine associations between asthma in offspring and ART use, adjusting for parental characteristics (age, education level, occupation type, BMI, asthma), smoking exposure, residence type, child sex, child age, and year of follow-up. Mediating effects were explored using longitudinal mediation structural equation modelling. Asthma was reported for 51 (4.2%) of the 1206 NC singletons (median [interquartile range] age 5 [4-5] years; 48.1% females) and 169 (5.2%) of the 3227 ART-conceived singletons (5 [5-5] years; 47.6% females). We found that risks of childhood asthma in singletons conceived by ART were, overall, similar to those of NC singletons before (odds ratio [OR], 1.25 [95% CI, 0.92-1.74]; P = 0.170) and after adjustment (adjusted OR [aOR], 0.66 [95% CI, 0.44-1.03]; P = 0.126). The results were similar in multiple sensitivity analyses, and there were no clear differences in asthma risks according to the method of ART. Mediation analysis revealed a significant positive indirect effect of neonatal intensive care unit (NICU) admission (standard path coefficient, b = 0.025, P < 0.05) and a negative indirect effect of breastfeeding (b = -0.012, P < 0.05) on the association between ART and asthma in singleton offspring. This study is limited to singletons only and cannot be generalized. The study is also limited by its retrospective observational single-centre nature and sample size. Mediation analyses were exploratory. Therefore, the findings need to be interpreted with caution. These findings can help infertile couples undergoing ART be reassured about the risk of childhood asthma in singleton offspring. Breastfeeding is recommended as a potentially feasible intervention to reduce the asthma risks in ART-conceived children who are at increased potential risk of asthma, such as those with NICU admissions. This work was supported by the Key Research and Development Program of Zhejiang Province (2021C03100), the National Key Research and Development Program of China (2021YFC2700603), and the Program for Key Subjects of Zhejiang Province in Medicine and Hygiene to Y. Z., the Zhejiang Province Natural Science Foundation (No. LQ22H040006) and the National Natural Science Foundation of China (No.82101759) to M.T., and the National Natural Science Foundation of China (No. 82201860) to J.Y. The authors declare no competing interests. ChiCTR2300069906.

Association of ambient air pollutant mixtures with IVF/ICSI-ET clinical pregnancy rates during critical exposure periods

Abstract STUDY QUESTION Does exposure to a mixture of ambient air pollutants during specific exposure periods influence clinical pregnancy rates in women undergoing IVF/ICSI-embryo transfer (ET) cycles? SUMMARY ANSWER The specific exposure period from ET to the serum hCG test was identified as a critical exposure window as exposure to sulfur dioxide (SO2) or a combination of air pollutants was associated with a decreased likelihood of clinical pregnancy. WHAT IS KNOWN ALREADY Exposure to a single pollutant may impact pregnancy outcomes in women undergoing ART. However, in daily life, individuals often encounter mixed pollution, and limited research exists on the effects of mixed air pollutants and the specific exposure periods. STUDY DESIGN, SIZE, DURATION This retrospective cohort study involved infertile patients who underwent their initial IVF/ICSI-ET cycle at an assisted reproduction center between January 2020 and January 2023. Exclusions were applied for patients meeting specific criteria, such as no fresh ET, incomplete clinical and address information, residency outside the 17 cities in the Sichuan Basin, age over 45 years, use of donor semen, thin endometrium (&amp;lt;8 mm) and infertility factors unrelated to tubal or ovulation issues. In total, 5208 individuals were included in the study. PARTICIPANTS/MATERIALS, SETTING, METHODS Daily average levels of six air pollutants (fine particulate matter (PM2.5), inhalable particulate matter (PM10), SO2, nitrogen dioxide (NO2), carbon monoxide (CO), and ozone (O3)) were acquired from air quality monitoring stations. The cumulative average levels of various pollutants were determined using the inverse distance weighting (IDW) method across four distinct exposure periods (Period 1: 90 days before oocyte retrieval; Period 2: oocyte retrieval to ET; Period 3: ET to serum hCG test; Period 4: 90 days before oocyte retrieval to serum hCG test). Single-pollutant logistic regression, two-pollutant logistic regression, Quantile g-computation (QG-C) regression, and Bayesian kernel machine regression (BKMR) were employed to evaluate the influence of pollutants on clinical pregnancy rates. Stratified analyses were executed to discern potentially vulnerable populations. MAIN RESULTS AND THE ROLE OF CHANCE The clinical pregnancy rate for participants during the study period was 54.53%. Single-pollutant logistic models indicated that for PM2.5 during specific exposure Period 1 (adjusted odds ratio [aOR] = 0.83, 95% CI: 0.70–0.99) and specific exposure Period 4 (aOR = 0.83, 95% CI: 0.69–0.98), and SO2 in specific exposure Period 3 (aOR = 0.92, 95% CI: 0.86–0.99), each interquartile range (IQR) increment exhibited an association with a decreased probability of clinical pregnancy. Consistent results were observed with dual air pollution models. In the multi-pollution analysis, QG-C indicated a 12% reduction in clinical pregnancy rates per IQR increment of mixed pollutants during specific exposure Period 3 (aOR = 0.89, 95% CI: 0.79–0.99). Among these pollutants, SO2 (33.40%) and NO2 (33.40%) contributed the most to the negative effects. The results from BKMR and QG-C were consistent. Stratified analysis revealed increased susceptibility to ambient air pollution among individuals who underwent transfer of two embryos, those with BMI ≥ 24 kg/m2 and those under 35 years old. LIMITATIONS, REASONS FOR CAUTION Caution was advised in interpreting the results due to the retrospective nature of the study, which was prone to selection bias from non-random sampling. Smoking and alcohol, known confounding factors in IVF/ICSI-ET, were not accounted for. Only successful cycles that reached the hCG test were included, excluding a few patients who did not reach the ET stage. While IDW was used to estimate pollutant concentrations at residential addresses, data on participants’ work locations and activity patterns were not collected, potentially affecting the accuracy of exposure prediction. WIDER IMPLICATIONS OF THE FINDINGS Exposure to a mixture of pollutants, spanning from ET to the serum hCG test (Period 3), appeared to be correlated with a diminished probability of achieving clinical pregnancy. This association suggested a potential impact of mixed pollutants on the interaction between embryos and the endometrium, as well as embryo implantation during this critical stage, potentially contributing to clinical pregnancy failure. This underscored the importance of providing women undergoing ART with comprehensive information to comprehend the potential environmental influences and motivating them to adopt suitable protective measures when feasible, thereby mitigating potential adverse effects of contaminants on reproductive health. STUDY FUNDING/COMPETING INTEREST(S) This work received support from the National Key Research and Development Program of China (No. 2023YFC2705900), the National Natural Science Foundation of China (Nos. 82171664, 81971391, 82171668), the Natural Science Foundation of Chongqing Municipality of China (Nos. CSTB2022NSCQ-LZX0062, CSTB2023TIAD-KPX0052) and the Foundation of State Key Laboratory of Ultrasound in Medicine and Engineering (No. 2021KFKT013). The authors report no conflicts of interest. TRIAL REGISTRATION NUMBER N/A.

Agalychnis callidryas: problemas y soluciones para su cría en cautiverio

Frogs of the genus Agalychnis and particularly A. callidryas, with distribution in Mexico, are widely poached for the pet trade, one of their main threats. In this sense, captive reproduction of amphibians is a viable alternative for conservation although it is not easy in species with the characteristics of this frog. In the present work, some complications that arise during captive reproduction of A. callidryas and their possible solutions are described, and treatments that allowed the solution of these problems are shown: bacterial infections, helminthic infections and skin tears. The use of their body patterns for the identification of individuals is also included. The objective is to provide alternatives that can be used in zoos, aquariums and other reproductive centers, as well as in breeding programs for the conservation of threatened anurans similar to this species.