Embryo Quality Research Articles

Eicosatrienoic acid enhances the quality of in vitro matured porcine oocytes by reducing PRKN-mediated ubiquitination of CISD2

Oocytes and early embryos are exposed to many uncontrollable factors that trigger endoplasmic reticulum (ER) stress during in vitro culture. Prevention of ER stress is an effective way to improve the oocyte maturation rate and oocyte quality. Increasing evidence suggests that dietary intake of sufficient n-3 polyunsaturated fatty acids (PUFAs) is associated with health benefits, particularly in the domain of female reproductive health. We found that supplementation of eicosatrienoic acid (ETA) during in vitro maturation (IVM) of oocyte significantly downregulated ER stress-related genes. Mitochondria-associated membranes (MAMs) are communications areas between the ER and mitochondria. Inositol 1,4,5-trisphosphate receptor (IP3R) is a key calcium channels in MAMs and, participates in the regulation of many cellular functions. Notably, the MAM area was significantly decreased in ETA-treated oocytes. CDGSH iron sulfur domain 2 (CISD2) is presents in MAMs, but its role in oocytes is unknown. ETA treatment significantly increased CISD2 expression, and siRNA-mediated knockdown of CISD2 blocked the inhibitory effect of ETA on IP3R. Transcriptomic sequencing and immunoprecipitation experiments showed that ETA treatment significantly decreased expression of the E3 ubiquitin ligase PRKN. PRKN induced ubiquitination and degradation of CISD2, indicating that the PRKN-mediated ubiquitin-proteasome system regulates CISD2. In conclusion, our study reveals the mechanism by which ETA supplementation during IVM alleviates mitochondrial calcium overload under ER stress conditions by decreasing PRKN-mediated ubiquitination of CISD2 and facilitating inhibition of IP3R by CISD2/BCL-2. This improves oocyte quality and subsequent embryo developmental competence prior to implantation.

Reproductive outcomes in fresh transfer cycles and antagonists with premature luteinizing and/or progesterone surge: a single center retrospective cohort study.

The optimal outcome of assisted reproductive technology is a successful live birth after fresh embryo transfer. However, the success pregnancy rate of fresh embryo transfer cycle in antagonist protocol is lower than that observed in other protocols. Despite the use of antagonists (GnRH-ant), the incidence of luteinizing hormone surge and elevated progesterone levels remain at approximately 5%-38%. Progesterone is widely recognized to exert adverse effects on fresh embryo transfer outcomes. This study aimed to investigate the impact of luteinizing hormone surge and progesterone levels on live birth rate following fresh embryo transfer and explore appropriate progesterone thresholds to enhance pregnancy outcomes. This retrospective cohort study included a total of 1,177 antagonist protocol cycles with fresh embryo transfer. The patients were divided into four groups based on the presence of premature LH surge and progesterone level on trigger day>1.5ng/ml. Then, the relationship between the variables and the pregnancy outcome was analyzed and compared in each group. The transient rise of luteinizing hormone did not impact pregnancy outcomes (P=0.345; P=0.3; P=0.787), in contrast to progesterone levels on the day of hCG administration (P=0.047*; P=0.015*; P=0.021*). In cases with luteinizing hormone surge, elevated progesterone levels were correlated with higher antral follicle count (AFC), and as progesterone levels increased, a greater quantity of oocytes and embryos were obtained. However, there was no statistically significant difference in pregnancy outcomes. In cases without luteinizing hormone surge, elevated progesterone levels led to significantly poorer pregnancy outcomes. Furthermore, the curve-fitting and threshold-effect analysis revealed a notable decline in live birth rates when progesterone exceeded or equaled 1.10ng/ml (OR, 0.25; 95% CI, 0.09-0.66; P = 0.005*). The GnRH-ant dosage addition should be carefully selected in flexible antagonist protocols. The presence of elevated progesterone levels may be associated with improved embryo quality when luteinizing hormone surge occurred. In the absence of a luteinizing hormone surge, progesterone levels showed a larger impact on the pregnancy outcome, and fresh embryo transfer should not be performed if the progesterone level on the day of hCG administration is higher than 1.10ng/ml.

Antimony exposure affects oocyte quality and early embryo development via excessive mitochondrial oxidation and dysfunction

Antimony (Sb) is a metalloid, widely presents in the environment and associates with human health. In this study, we aimed to decipher whether Sb exposure is harmful to female reproduction and explore the underlying mechanisms. The ICR mice were exposed to 0, 5, 10, and 20 mg/kg acetate potassium Sb tartrate trihydrate by intraperitoneal injection for 10 days, then mouse oocytes were collected for further analysis. We first found a significant decrease in the proportion of MII-stage oocytes obtained from supernumerary ovulation in the fallopian tubes and early embryo development under Sb treatment. Then a series of tests showed Sb affects oocyte maturation by damaging the cytoskeleton of microtubule and actin. Moreover, the abnormal distribution of cortical granules and their component Ovastacin in oocytes, combined with reduced expression levels of Juno, affected sperm-oocyte binding and led to fertilization failure. Based on the sequencing results and experimental validation, it was demonstrated that Sb exposure impairs mitochondrial distribution and membrane potential, elevated levels of mitochondrial superoxide, finally caused energy supply deficits. Mitochondrial damage in oocytes after Sb exposure results in the excessive oxidative stress and early apoptosis. Taken together, these data suggest that Sb exposure decreases oocyte quality and female fertilization ability by impairing mitochondrial function and redox perturbation.

Impact of sperm sex sorting on sperm quality and in vitro embryo production in bovine

Increasing evidence suggests that environmental exposures can modify epigenetic marks in the germline, leading to the transmission of abnormal post-fertilization sperm epigenetic indicators and affecting embryonic development. Given the pivotal role of sperm cells in determining embryo quality, there is growing interest in understanding the potential effects of sperm sex sorting on embryo quality. This study aimed to investigate the impact of bovine sperm sexing on in vitro embryo production (IVP) and to associate molecular aspects of embryos analysis. Frozen semen samples from five Nellore bulls were used, with each bull contributing unsexed sperm (conventional semen – CV treatment) and female and male sexed sperm pooled after thawing (SX treatment). First, semen quality was assessed, including motility, morphology, acrosome integrity, and chromatin integrity to denaturation. Then, IVP was carried out, focusing on embryonic production and developmental kinetics. In the third experiment, embryo quality was evaluated by examining the gene expression of key markers (OCT4, NANOG, DNMT3A, TET1, and Fematrin-1) and the methylation pattern of the Satellite-1 and α-Satellite genes in blastocysts. Differences between CV and SX semen were only observed in motility, which was lower in SX compared with CV (P < 0.05). Although cleavage was similar, the SX groups showed lower blastocyst production than CV (P < 0.05). Of the genes evaluated, only NANOG showed high expression in the CV blastocysts compared with the SX blastocysts, but the methylation pattern revealed no differences. In conclusion, sex sorting markedly affects sperm motility and in vitro embryo production but showed no significant impact on embryo quality.

Environmental high temperature affects pre-implantation embryo development by impairing the DNA repair ability

Environmental high temperature poses a significant threat to human health, however, limited information is available for understanding the relationship between the hot weather and infertility. This study aims to assess the adverse effect of the hot weather to early embryonic cells. Our results indicated that environmental high temperature exposure could cause the decline of early embryo quality and implantation ability. In detail, it led to early embryonic development retardation, embryo degeneration rate increased, the rate of blastocyst and hatching decreased, and reduced the number of implants. And the finding also the impairment of environmental high temperature on early embryonic cells may be due to oxidative damage of DNA caused by ROS, while BER repair ability is decreased, failing to repair oxidative damage of DNA in time, resulting in a large number of early embryonic apoptosis. The work underscored that pregnant women should stay away from high-temperature environments.

Assisted oocyte activation significantly improves zygote formation, cleavage, and implantation rates in patients with a history of fertilization failures.

Fertilization check performed at the 18th hour following classic in vitro fertilization procedure (IVF) or intracytoplasmic sperm injection (ICSI) is a critical stage in assisted reproduction. The success of the treatment is significantly reliant on the quantity of zygotes exhibiting two pronuclei. Consequently, low fertilization rates or complete fertilization failure are highly undesirable outcomes for both patients and reproductive specialists. Applying additional calcium ionophore for oocyte activation subsequent to ICSI may offer benefits and potentially enhance treatment outcomes, particularly for patients who have experienced low or absent fertilization rates (FR) in previous treatment cycles. The aim of the study is to evaluate the efficacy of Ca2+ ionophore application for oocyte activation. A retrospective analysis of 924 oocytes obtained from 120 patients who underwent ICSI cycles with a history of low or no fertilization as a result of previous unsuccessful treatment rounds. The next ART cycle followed with additional oocyte Ca2+ ionophore activation applied in 57 of the cases in order to optimize the treatment process (Group 1), and 63 patients were included and their outcomes followed as a control group (Group 2).We conducted a comparative analysis of results in both groups. The study's primary outcomes encompassed fertilization, cleavage embryo quality, blastocyst rate, and established clinical pregnancies. At day 1fertilization check we had 274/386 zygotes (71%FR) in group 1 and 132/410 in group 2 (32.2%FR), (P<0.0001). Twenty-two (34.9%) cycles in group 2 resulted in total fertilization failure (TFF). At the cleavage stage top-quality embryos from group 1 were significantly higher (P = 0.0021) in comparison to group 2. Forty-eight embryo transfers (ET) were performed in group 1 resulting in 41.67% clinical pregnancies versus 33 ET and only 4 pregnancies (12.12%) for group 2 (P = 0.0044). The results confirm the appropriateness of assisted oocyte activation as an additional method in cases of previous fertilization failure cycles.

Impact of Amphiregulin on Oocyte Maturation and Embryo Quality: Insights from Clinical and Molecular Perspectives

AbstractIdentifying non-invasive biomarkers which can predict the outcome of intracytoplasmic sperm injection (ICSI) is crucial, particularly in Germany where the challenges are intensified by the Embryo Protection Act. Recent research has highlighted biomarkers within the epidermal growth factor (EGF) family as central to follicular processes, although their predictive utility remains a subject of debate in the literature. Therefore, the primary objective of this study was to investigate the significance of amphiregulin concentrations in follicular fluid and gene expression in mural granulosa cells on oocyte maturation, fertilization, and embryo quality.A total of 33 women were recruited at the University Clinic of Saarland Fertility Center (Homburg, Germany). Follicular fluid aspiration consisted of single/individual aspiration of follicles, enabling a 1 : 1 correlation with retrieved oocytes. Follicular fluid and mural granulosa cell samples from 108 oocytes were analyzed. Amphiregulin levels were determined with enzyme-linked immunosorbent assay, while gene expression was analyzed with the StepOnePlus Real-Time PCR System using TaqMan Fast Advanced Master Mix assays.Results showed that amphiregulin concentrations affect oocyte maturation, fertilization, and embryo quality, while luteinizing hormone concentrations influence oocyte maturation, with significant differences identified between fertilized/unfertilized and good/poor embryo groups. Amphiregulin expression significantly impacts oocyte maturation, with downregulation observed in immature oocytes, while luteinizing hormone/chorionic gonadotropin receptor expression showed no significant differences between groups and did not influence maturation, fertilization, or embryo quality.These findings are very important for advancing infertility treatment, especially in Germany. The results for amphiregulin may provide prognostic insights which could be useful when selecting viable oocytes and embryos. This research underscores the importance of non-invasive biomarkers for optimizing ICSI outcomes and potentially enhancing the success rates of assisted reproductive technology.

Does it affect the live birth rates to have a maximum endometrial thickness of 7, 8, or 9 mm in in-vitro fertilization-embryo transfer cycles?

To assess the effect of endometrial thickness (EMT) on live birth rates (LBR) in women with endometrial lining between 7.0-9.9 mm. This retrospective cohort study included women who underwent fresh and frozen embryo transfers between 2008 and 2018, grouped according to their maximum EMT; group 1, 7.0-7.9 mm; group 2, 8.0-8.9 mm; and group 3, 9.0-9.9 mm and underwent blastocyst transfer. The study included 7,091 in-vitro fertilization cycles: 1,385 in group 1, 3,000 in group 2, and 2,706 in group 3. The combined LBR was 22.2%. The mean age of women at oocyte retrieval day was 36.7±4.5 years. There was no difference in female age at oocyte retrieval or in the quality of embryos transferred between the three groups. Group 1 had more diagnoses of diminished ovarian reserve (25.8% vs. 19.5% and 19.1%; p<0.001) and less male factor infertility compared with group 2 and 3, respectively (25.0% vs. 28.8% and 28.5%; P=0.024). LBR was higher with increasing endometrial thickness, group 2 vs. group 1 (22.0% vs. 17.4%; P=0.0004), group 3 vs. group 1 (25.0% vs. 17.2%; p<0.001), and group 3 vs. group 2 (25.0% vs. 22.0%; P=0.008). After controlling for confounding factors, these three groups did not differ in LBR (group 1 vs. group 2, odds ratio [OR], 1.08; 95% confidence interval [CI], 0.83-1.4; P=0.54 and group 1 vs. group 3, OR, 1.16; 95% CI, 0.90-1.51; P=0.24). Live birth rates in women with endometrial thickness between 7.0-9.9 mm were not affected by different cut-offs when blastocyst transfer was performed.

Subendometrial blood flow detected by Doppler ultrasound associates with pregnancy outcomes of frozen embryo transfer in patients with thin endometrium.

Multiple factors have been shown to influence the rate of clinical pregnancy after FET in IVF treatment, including embryo quality, synchronization of embryo and endometrium, and endometrial receptivity (ER). The subendometrial blood flow conditions could also contribute potentially major effects toward the establishment and maintenance of pregnancy. We conducted a retrospective cohort study to examine the correlation between subendometrial blood flow, as determined by Doppler ultrasound, and pregnancy outcomes in IVF patients with a thin endometrium (endometrium thickness [EMT] ≤ 0.7cm). This was a retrospective cohort study conducted at a university-affiliated reproductive hospital from January 2017 to April 2023. The EMT and subendometrial blood flows were assessed using transvaginal color Doppler ultrasound and evaluated by experienced clinical ultrasound physicians on the endometrial transformation day. The pregnancy outcomes were followed up and documented in clinical medical records through the IVF cohort study at our center. In the patients with 0.5cm ≤ EMT ≤ 0.7cm, the embryo implantation rate was statistically significant increased in the patients with the presence of subendometrial blood flow (OR 1.484; 95% CI, 1.001-2.200; P = 0.049; aOR 1.425; 95% CI, 1.030-2.123; P = 0.003). Patients with discernible subendometrial blood flow have superior live birth (P = 0.028), clinical pregnancy (P = 0.049), and embryo implantation (P = 0.027) compared to the patients without subendometrial blood flow when the EMT is ≤ 0.7cm. The presence of subendometrial blood flow detected by ultrasound was positively associated with successful embryo implantation and favorable pregnancy outcomes in patients with thin endometrium undergoing FET.

Intraovarian PRP injection improves oocyte quality and early embryo development in mouse models of chemotherapy-induced diminished ovarian reserve.

Intraovarian injection of platelet-rich plasma (PRP) has been recently proposed, with encouraging results to provide an alternative option to patients diagnosed with POR or POI. However, the broad spectrum of PRP effects on the reproductive function and the mechanisms of action in follicular activation, response to stimulation, and embryo quality have not yet been studied. In this study, we first induced poor ovarian reserve (POR) and premature ovarian insufficiency (POI) ovarian phenotypes in CD1 mice undergoing PRP or sham intraovarian injection. PRP administration reduced those alterations induced by chemotherapy in ovarian stroma and follicle morphology in both the POR and POI conditions. After ovarian stimulation, we found that PRP did not modify the MII-oocyte yield. Nevertheless, the amount of obtained 2-cell embryos and fertilization rate were increased, being especially relevant for the POI model. Further in vitro embryo culture led to improved blastocyst formation rates and higher numbers of good quality blastocysts in PRP vs. sham females in both the POR and POI conditions. These positive results of PRP injection were also validated in the C57Bl/6 stain. Altogether, our findings suggest a possible effect on oocyte and embryo quality. This effect is likely due to the increase of local paracrine signaling through the released growth factors in PRP-treated ovaries.

229 Awardee Talk: The quest for an ideal in-field embryo evaluation tool in cattle: Opportunities and challenges

Abstract Accurately evaluating bovine embryos to select the most viable ones for transfer and successful pregnancy has been a challenge since embryo transfer began in cattle over six decades ago. Despite more than 3,100 peer-reviewed studies on bovine embryo evaluation published in PubMed from 1964 to 2023, there is still no consensus or gold standard method for assessing their developmental potential. The evaluation “problem” in assisted reproduction extends beyond embryos to gametes as well. Numerous microscopic techniques (e.g., differential interference contrast, electron, fluorescent, time-lapse, and artificial intelligence-based microscopy) and non-microscopic methodologies (e.g., genomics, transcriptomics, epigenomics, proteomics, metabolomics, and nuclear magnetic resonance) have been tested to find an embryo evaluation technique superior to morphological evaluation. While many of these research tools can accurately determine embryo quality and viability, most are invasive, expensive, labor-intensive, technically complex, and time-consuming, making them impractical for in-field embryo evaluation. Employing complex methods like RNA sequencing, SNP chips, mass spectrometry, and multiphoton microscopy at thousands of embryo production and collection facilities is unrealistic, no matter how accurate they may be. Therefore, future research should focus on innovating field-friendly, simple benchtop tests using existing findings, particularly from omics-based research methodologies. Time-lapse monitoring and artificial intelligence-based automated image analysis also potentially provide accurate embryo evaluation. However, further research is necessary to develop economically feasible options for in-field applications. Our recent work has examined possible applications of nuclear magnetic resonance (NMR) imaging, label-free microscopy, particularly holographic microscopy techniques like gradient light interference microscopy (GLIM), spatial light interference microscopy (SLIM), and multiphoton holographic microscopy in IVF labs. The ideal embryo evaluation tool should be accurate, objective, non-invasive, affordable, and simple enough for widespread adoption by embryo production and collection facilities worldwide. Under current circumstances, it is doubtful that transcriptomics, proteomics, metabolomics, GLIM, SLIM, NMR, multiphoton holographic microscopy, and similar techniques can be directly used for in-field embryo evaluation. However, biomarkers identified by these methodologies could be used for in-field embryo evaluation by devising simple, affordable benchtop techniques. Further investigation on time-lapse microscopy (TLM) and AI-based automated image processing may make these methods more affordable and potentially adoptable by the masses. There is great potential for these methodologies to become widely used, provided they can be made more economically viable. In conclusion, the next generation of microscopy capable of providing objective markers for gamete and embryo quality is on the horizon.

7 Role of the preovulatory follicle in oocyte metabolic and developmental competence for pregnancy

Abstract A majority of bovine pregnancy loss occurs in the first 7 d of gestation. The oocyte is a key driver of embryo viability during this time, and its competence for embryo development and pregnancy establishment is greatly influenced by the surrounding microenvironment during oocyte maturation. Assisted reproductive technologies allow for more efficient propagation of superior genetics and improved reproductive management of the cattle herd; however, many assisted reproductive technologies alter the normal microenvironment of the maturing oocyte and negatively impact oocyte competence for early embryo development. For example, oocytes induced to ovulate from preovulatory follicles of lesser physiological maturity (decreased estradiol production and smaller diameter) have reduced blastocyst development, poorer embryo quality, and a hastened rate of embryo development compared with those ovulated from follicles of greater maturity. Our overarching hypothesis is that the intrafollicular milieu of physiologically mature preovulatory follicles provides the necessary microenvironment to metabolically program maturing oocytes for successful embryo development. Studies by our team have demonstrated that the follicular fluid metabolite milieu during in vivo oocyte maturation differs between preovulatory follicles of greater and lesser maturity and that such differences have direct effects on the maturing oocyte. Numerous follicular fluid metabolites involved in glucose and amino acid metabolism were more abundant in preovulatory follicles of greater versus lesser physiological maturity. Additionally, abundance of transcripts encoding glycolytic enzymes was higher in cumulus cells from follicles of greater maturity, and both ATP and levels of key transcripts in the mitochondrial respiratory chain were higher in oocytes collected from preovulatory follicles of greater versus lesser maturity. We supplemented in vitro maturing oocytes with follicular fluid from preovulatory follicles of greater or lesser maturity to determine causality of differing follicular fluid milieu on cumulus-oocyte metabolism and oocyte competence for early embryo development. Follicle maturity status had robust effects on cumulus-oocyte complex consumption or production of key metabolites in glucose, purine, and amino acid metabolism pathways during oocyte maturation. We also observed hastened development rate in embryos arising from in vitro matured oocytes supplemented with lesser maturity follicular fluid, mirroring past in vivo findings by others. Furthermore, RNA-sequencing of resulting blastocysts of identical stage and quality suggested more efficient metabolism and Wnt/β-catenin signaling in blastocysts derived from oocytes matured in preovulatory follicular fluid from greater maturity follicles. In contrast, transcriptome profiles of blastocysts from oocytes matured in preovulatory follicular fluid from lesser maturity follicles indicated oxidative stress and dysregulated cell division. These recent findings by our lab indicate that impacts of follicle maturity and the resulting follicular fluid microenvironment on the maturing oocyte withstand throughout early embryo development and hold promising roles to improve reproductive efficiency in cattle.

Telomeric RNAs, TERRA, as a Potential Biomarker for Spermatozoa Quality.

Characterization of long non-coding telomeric repeat-containing RNAs in sperm of normozoospermic and oligoasthenozoospermic men as new biomarker of idiopathic male infertility. We conducted an observational prospective study with two groups of men with normal or orligoasthenozoospermic spermiogram, aged 40 and above. Fertility parameters were analyzed in men undergoing intracytoplasmic sperm injection with donor oocytes, to avoid the female factor. Telomeric RNAs and telomere length were measured by quantitative fluorescent in situ hybridization. Data from seminal parameters and in-vitro fertilization were assessed according to IVIRMA protocols. Patients with oligoasthenozoospermia, who had worse seminal parameters, also obtained embryos with lower inner-cell-mass quality (p = 0.04), despite using donor oocytes. While mean levels of telomeric RNAs were similar for both groups, the percentage of spermatozoa with more than 3 foci was higher in oligoasthenozoospermic men (p = 0.02). Regarding telomere length, oligoasthenozoospermic men had shorter mean, a higher accumulation of short telomeres (15th percentile; p = 0.03) and a lower percentage of very-long telomeres (85th percentile; p = 0.01). Finally, a positive correlation was found between telomeric-RNAs intensity and total progressive motility in the spermatozoa of normozoospermic patients (r = 0.5; p = 0.03). Telomeric parameters were altered in the spermatozoa of the oligoasthenozoospermic group, which also showed lower quality embryos. Interestingly, in the normozoospermic group, a correlation was found between progressive motility and telomeric RNA levels, suggesting that they could be a good biomarker of sperm quality. Further studies are required to confirm these results and translate them into the clinical practice.Trial registration number: 1711-MAD-109-CB, 07/07/2021.

A two-year plant-based diet alters the fatty acid profile and enzymatic and non-enzymatic lipid metabolites, in the eggs and fry of female rainbow trout

To promote sustainable aquaculture, plant-based ingredients are increasingly being used to replace fish meal (FM) and fish oil (FO) in diets, affecting critical fatty acid (FA) profiles in broodstock diets. These profiles are essential for reproduction and offspring survival. The absence of traditional fish-derived nutrients results in an altered FA composition, particularly a reduction in omega-3 long-chain polyunsaturated fatty acids (LC-PUFAs). This change is detrimental to trout reproduction over the long term. Current research gaps, especially regarding FA profiles and related metabolites, limit our understanding of the reproductive consequences of omega-3 deficient plant-based diets. To fill this knowledge gap, a two-year trial was conducted to compare the reproductive effects (spawning performances, egg quality, embryo development) of two diets on rainbow trout females: a plant-based diet containing linseed (10.8 %), palm (5 %), and sunflower oils (2.7 %), and a commercial diet with high FM (27 %) and FO (12.1 %) content. Both diets were similar in terms of protein (44 %), lipids (23.4 %), and energy (25 kJ/g). The study spanned from initial feeding to first reproduction, and fatty acid profiles and metabolites (selected enzymatic and non enzyamatic metabolites from FA) were examined in the broodstock diet, female muscle tissue, eggs, and fry. Trout fed a plant-based diet showed reduced reproductive traits such as lower weight and fecundity, but egg and embryo quality remained high. Survival and hatching rates were similar to those on commercial diets. The study revealed for the first time the critical role of maternal diet in the generation of lipid metabolites in the offspring, particularly DHA (F4-neuroprostane, maresin, 14-HDoHE) and AA (F2-isoprostane, prostaglandin E and F2, 8–12- 15-HETEs) oxylipins, highlightingthe complex dietary influences on fish reproduction. This work underlines that a plant-based diet significantly affects the the reproductive performance of broodstock trout by altering fatty acid profiles and metabolite levels and highlights the delicate balance between dietary components, oxylipins, and reproductive functions in fish, suggesting the need for more comprehensive studies of their interrelated effects.

Letrozole co-treatment in an antagonist protocol for overweight women undergoing IVF treatment: a retrospective study

BackgroundOverweight women undergoing IVF treatment have lower success rates. Letrozole, an aromatase inhibitor, has been used as an adjunct for IVF treatment, but its specific effects in overweight women have not been investigated. This study was to explore the effects of letrozole co-treatment in an antagonist protocol for overweight infertile women undergoing IVF treatment.MethodsThis retrospective cohort study included overweight infertile women who underwent IVF/ICSI treatment and fresh embryo transfer (ET), with or without letrozole co-treatment in an antagonist protocol, from 2007 to 2021 at Shanghai Ninth People’s Hospital (Shanghai, China). A total of 704 overweight infertile women were included: 585 women were in the antagonist group, and 119 women were in the letrozole co-treatment group. The primary outcome was the live birth rate after fresh ET. Propensity score-based patient-matching was employed to balance the covariates between the groups. Multivariate logistic regression analysis was also performed to estimate odds ratio (OR) and 95% confidence interval (CI) for association of letrozole co-treatment and the live birth outcome.ResultsLetrozole co-treatment induced significant changes in hormonal profile on the trigger day. The letrozole group exhibited a decrease in the total number of follicles compared to the antagonist group, but a higher proportion of large follicles at oocyte retrieval (P < 0.05). The quantity and quality of embryos were comparable between the two groups (P > 0.05). The letrozole co-treatment group had a significantly higher live birth rate than the control group (38.7% vs. 22.6%, P = 0.026). With multivariate logistic regression analysis, letrozole co-treatment was associated with higher odds of live birth after adjusting for potential confounding factors (adjusted OR = 2.00, 95% CI = 1.17–3.39, P = 0.011). Letrozole presented no significant associations with obstetrical or neonatal complications (P > 0.05).ConclusionLetrozole co-treatment in an antagonist protocol may offer potential benefits for overweight infertile women undergoing IVF treatment. Further research is warranted to validate these findings and explore the broader implications for letrozole co-treatment.

Pentose phosphate pathway inhibition during in vitro maturation substantially affects the metabolism of bovine COCs and blastocysts

Glucose metabolism is widely examined in terms of its effect on oocytes and embryos quality. There are two main pathways of glucose metabolism – glycolysis and pentose phosphate pathway (PPP). The glycolytic pathway allows for energy production in the form of ATP and metabolites such as pyruvate and lactate, whereas PPP activity generates NADPH as well as ribose 5-phosphate, a precursor for the synthesis of nucleotides. The aim of the present experiment was the selective inhibition of either glycolysis or PPP during in vitro maturation of bovine cumulus-oocyte complexes (COCs) to demonstrate, how it affects COCs and further embryos with regard to selected lipidomic and metabolomic aspects. Inhibitors of glycolysis (IO) or PPP (DHEA) were applied during IVM, and the control group was matured under standard conditions. A set of COCs from each group was fertilized and obtained embryos were cultured to the blastocyst stage. ATP level was measured in oocytes, relative mRNA level of selected genes involved in energy metabolism was measured in cumulus cells (CC; real time PCR), lipid droplets parameters were evaluated in oocytes and CC whereas metabolome and lipidome (mass spectrometry) were evaluated in oocytes, CC and blastocysts as well. The experiment shows that glycolysis inhibition during IVM affects mainly CC with no effect in oocytes. It allows to maintain the good developmental potential of oocytes and no negative effect of blastocysts quality and quantity is observed. In contrary, PPP inhibition negatively affects metabolic and lipidomic parameters of both oocyte and CC, which further decreases blastocyst rate and quality. It is therefore concluded that PPP is the most crucial pathway of glucose metabolism for COC developmental potential.

Human sperm RNA in male infertility.

The function and value of specific sperm RNAs in apparently idiopathic male infertility are currently poorly understood. Whether differences exist in the sperm RNA profile between patients with infertility and fertile men needs clarification. Similarly, the utility of sperm RNAs in predicting successful sperm retrieval and assisted reproductive technique (ART) outcome is unknown. Patients with infertility and fertile individuals seem to have differences in the expression of non-coding RNAs that regulate genes controlling spermatogenesis. Several RNAs seem to influence embryo quality and development. Also, RNA types seem to predict successful sperm retrieval in patients with azoospermia. These findings suggest that sperm RNAs could influence decision-making during the management of patients with infertility. This evidence might help to identify possible therapeutic approaches aimed at modulating the expression of dysregulated genes in patients with infertility. Performing prospective studies with large sample sizes is necessary to investigate cost-effective panels consisting of proven molecular targets to ensure that this evidence can be translated to clinical practice.

Adolescent exposure to tris(1,3-dichloro-2-propyl) phosphate (TCPP) induces reproductive toxicity in zebrafish through hypothalamic-pituitary-gonadal axis disruption

Tris(1,3-dichloro-2-propyl) phosphate (TCPP), a prevalent organophosphorus flame retardant in aquatic environments, has raised significant concerns regarding its ecological risks. This study aims to explore the impacts of TCPP on the reproductive functions of zebrafish and delineate its gender-related toxic mechanisms. By assessing the effects on zebrafish of 10 mg/L TCPP exposure from 30 to 120 days post-fertilization (dpf), we thoroughly evaluated the reproductive capability and endocrine system alterations. Our findings indicated that TCPP exposure disrupted gender differentiation in zebrafish and markedly impaired their reproductive capacity, resulting in decreased egg laying and offspring development quality. Histological analyses of gonadal tissues showed an abnormal increase in immature oocytes in females and a reduction in mature sperm count and spermatogonial structure integrity in males, collectively leading to compromised embryo quality. Additionally, molecular docking results indicated that TCPP showed a strong affinity for estrogen receptors, and TCPP-treated zebrafish exhibited imbalanced sex hormones and increased estrogen receptor expression. Alterations in genes associated with the hypothalamic-pituitary-gonadal (HPG) axis and activation of the steroidogenesis pathway suggested that TCPP targets the HPG axis to regulate sex hormone homeostasis. Tamoxifen (TAM), as a competitive inhibitor of estrogen, exhibited a biphasic effect, as evidenced by the counteraction of TCPP-induced effects in both male and female zebrafish after TAM addition. Overall, our study underscored the gender-dependent reproductive toxicity of TCPP exposure in zebrafish, characterized by diminished reproductive capacity and hormonal disturbances, likely due to interference in the HPG axis and steroidogenesis pathways. These findings emphasize the critical need to consider gender differences in chemical risk assessments for ecosystems and highlight the importance of understanding the mechanisms underlying the effects of chemical pollutants on the reproductive health of aquatic species.

The Role of the Endometrium in Implantation: A Modern View.

According to the current data, the endometrium acts as a "sensor" of embryo quality, which promotes the implantation of euploid embryos and prevents the implantation and/or subsequent development of genetically abnormal embryos. The present review addresses the nature of the "sensory function" of the endometrium and highlights the necessity for assessing its functional status. The first section examines the evolutionary origin of the "sensory" ability of the endometrium as a consequence of spontaneous decidualization that occurred in placental animals. The second section details the mechanisms for implementing this function at the cellular level. In particular, the recent findings of the appearance of different cell subpopulations during decidualization are described, and their role in implantation is discussed. The pathological consequences of an imbalance among these subpopulations are also discussed. Finally, the third section summarizes information on currently available clinical tools to assess endometrial functional status. The advantages and disadvantages of the approaches are emphasized, and possible options for developing more advanced technologies for assessing the "sensory" function of the endometrium are proposed.

The impact of blastocyst grade on singleton birth weight in fresh IVF‒ET cycles in ART: a retrospective study

BackgroundThe positive correlation between embryo quality and pregnancy outcomes has been confirmed in many studies, but there are few on the impact of embryo quality on neonatal weight, especially among neonates from fresh IVF‒ET cycles in ART. Therefore, this study aimed to compare the birth weights of infants from different blastocyst grades in fresh IVF-ET cycles and explore related factors affecting birth weight.MethodsThe main outcome measure was singleton birth weight. A total of 1301 fresh cycles of single blastocyst transplantation and single live birth profiles were retrospectively analyzed and divided into four groups according to blastocyst quality: the excellent group (grade AA), which included 170 cycles; the good group (grade AB/BA), which included 312 cycles; the average group (grade BB/CA/AC), which included 559 cycles; and the poor group (grade BC/CB), which included 260 cycles. The relationships among cystic cavity expansion, endocytic cell mass, ectodermal trophoblast cell grade, and birth weight were studied. Multiple linear regression analysis was performed to investigate the relationship between blastocyst quality and neonatal birth weight and logistic regression for the risk factors for low birth weight newborns.ResultsWith decreases in the blastocyst quality, including ICM, TE quality, and embryo expansion stage, birth weight declined, and Z scores correspondingly decreased. After adjusting for confounders, the average and poor groups (P = 0.01 and P = 0.001, respectively) and blastocysts with TE grade C (P = 0.022) resulted in singletons with lower birth weight. Additionally, the poor group and blastocysts with Grade C TEs had a greater chance of leading to low birth weight infants compared with the other groups.ConclusionOur findings indicated that excellent and good-grade blastocyst transplantation could achieve better pregnancy outcomes and that average and poor-grade blastocyst transplantation, especially with grade C TEs, were associated with single birth weight loss. No association was found between the embryo expansion stage or ICM quality and neonatal birth weight.