Assisted Reproductive Outcomes Research Articles

The clinical effectiveness of the Mind/Body Program for Infertility on wellbeing and assisted reproduction outcomes: a randomized controlled trial in search for active ingredients.

Does the Mind/Body Program for Infertility (MBPI) perform better, due to certain distinctive elements, than a partly matched support group in improving the wellbeing and medically assisted reproduction (MAR) outcomes of women with elevated distress levels in a clinical setting? While robust enhancements occurred in the wellbeing overall, the cognitive behavioural and formalized stress management elements of the MBPI allowed a significantly stronger improvement in trait anxiety, but not in other mental health and MAR outcomes, compared with a support group. Mind-body psychological programmes adjacent to MAR have been found to improve women's mental states and possibly increase chances of pregnancy. However, not enough is known about the programme's effectiveness among patients with elevated distress levels in routine clinical settings, nor is it clear which of its particular ingredients are specifically effective. A pre-post design, single-centre, randomized controlled trial was performed between December 2019 and October 2022 (start and end of recruitment, respectively). The sample size (n = 168) was calculated to detect superiority of the MBPI in improving fertility-related quality of life. Randomization was computer-based, with random numbers concealing identities of patients until after allocation. The trial was conducted at a large university teaching hospital. A total of 168 patients were randomly assigned to the mind-body (MBPI) group (n = 84) and the fertility support (FS) control group (n = 84). Patients received a 10-week, 135-min/week group intervention, with the FS group following the same format as the MBPI group, but with a less restricted and systematic content, and without the presumed effective factors. The number of patients analysed was n = 74 (MBPI) and n = 68 (FS) for post-intervention psychological outcomes, and n = 54 (MBPI) and n = 56 (FS) for pregnancy outcomes at a 30-month follow-up. Significant improvements occurred in both groups in all psychological domains (adjusted P < 0.001), except for treatment-related quality of life. Linear mixed-model regression analysis did not reveal significantly greater pre-post improvements in the MBPI group than in the FS group in fertility-related quality of life (difference in differences (DD) = 4.11 [0.42, 7.80], d = 0.32, adjusted P = 0.124), treatment-related quality of life (DD = -3.08 [-7.72, 1.55], d = -0.20, adjusted P = 0.582), infertility-specific stress (DD = -2.54 [-4.68, 0.41], d = -0.36, adjusted P = 0.105), depression (DD = -1.16 [3.61, 1.29], d = -0.13, adjusted P = 0.708), and general stress (DD = -0.62 [-1.91, 0.68], d = -0.13, adjusted P = 0.708), but it did show a significantly larger improvement in trait anxiety (DD = -3.60 [-6.16, -1.04], d = -0.32, adjusted P = 0.042). Logistic regression showed no group effect on MAR pregnancies, spontaneous pregnancies, or live births. The follow-up only covered MAR-related medical outcomes and no psychological variables, and their rates were not equal in the two groups. Biological factors other than age, aetiology, and duration of infertility may have confounded the study results. Loss to follow-up was between 5% and 10%, which may have led to some bias. The psychologically and medically heterogeneous sample, the normal clinical setting and the low attrition rate all raise the external validity and generalizability of our study. The MBPI works not only in controlled conditions, but also in routine MAR practice, where it can be introduced as a cost-effective, low-intensity psychological intervention, within the framework of stepped care. More studies are needed to further identify its active ingredients. The authors received no financial support for the research, authorship, and/or publication of this article. The authors have no conflict of interest to disclose. ClinicalTrials.gov NCT04151485. 5 November 2019. 15 December 2019.

Mitochondria as therapeutic targets in assisted reproduction.

Mitochondria are essential organelles with specialized functions, which play crucial roles in energy production, calcium homeostasis, and programmed cell death. In oocytes, mitochondrial populations are inherited maternally and are vital for developmental competence. Dysfunction in mitochondrial quality control mechanisms can lead to reproductive failure. Due to their central role in oocyte and embryo development, mitochondria have been investigated as potential diagnostic and therapeutic targets in assisted reproduction. Pharmacological agents that target mitochondrial function and show promise in improving assisted reproduction outcomes include antioxidant coenzyme Q10 and mitoquinone, mammalian target of rapamycin signaling pathway inhibitor rapamycin, and nicotinamide mononucleotide. Mitochondrial replacement therapies (MRTs) offer solutions for infertility and mitochondrial disorders. Autologous germline mitochondrial energy transfer initially showed promise but failed to demonstrate significant benefits in clinical trials. Maternal spindle transfer (MST) and pronuclear transfer hold potential for preventing mitochondrial disease transmission and improving oocyte quality. Clinical trials of MST have shown promising outcomes, but larger studies are needed to confirm safety and efficacy. However, ethical and legislative challenges complicate the widespread implementation of MRTs.

P-548 An investigation into the mechanisms of active enhancers regulating the initial lineage differentiation in mammals

Abstract Study question What role do active enhancers play in determining the fate of early embryonic cells? Are there differences in active enhancers between mouse and human blastocysts? Summary answer Tissue-specific active enhancers in inner cell mass (ICM) and trophoblast (TE) are essential for blastocyst formation, regulating gene expression through interactive loops with promoters. What is known already At the two-cell stage of nuclear transfer embryos, the absence of interaction between Zscan4d promoter and distal active enhancer leads to reduced Zscan4d expression, resulting in lower developmental rates. Overexpressing the demethylase Kdm4d alleviates reprogramming-resistant H3K9me3 modification, restoring the structural loop between the Zscan4d promoter and active enhancer and enhancing developmental efficiency. These findings strongly suggest a close association between the formation of regulatory chromatin interactions in early embryonic development and histone modifications. Indicating active enhancers marked with H3K27ac may regulate differentiation gene expression through distal interactions in blastocyst. Study design, size, duration Analyze variations in histone H3K27ac modification, chromatin accessibility, higher-order structures, and transcriptomes during mouse embryo differentiation from 8-cell stage to ICM and TE formation. Identify tissue-specific active enhancers in ICM and TE, validate their gene expression regulation, explore their role in early differentiation-associated genes in mice. Compare and clarify similarities and differences in enhancer regulation mechanisms during early embryo development in humans and mice. Participants/materials, setting, methods The 8-cell, morula and blastocyst stage embryos were collected from the oviducts and uterus of super-ovulated female ICR mice mated with male mice. The clinical human blastocysts were collected and separated the ICM and TE. Low-input multi-omics sequencing, including Cut-Tag, ATAC-seq, Hi-C, and RNA-seq, was performed on the embryonic samples. After analyzing and identifying active enhancers, tissue-specific ones were selected and validated for their impact on gene expression via CRISPR-Cas9 knockout in embryos. Main results and the role of chance Inhibiting ep300 expression in mouse embryos through siRNA interference reduces blastocyst formation from 90% to approximately 40%. Introducing the small molecule inhibitor A485 at the 8-cell stage, blocking the active center of CBP/p300 enzymes, completely arrests embryos at the morula stage, preventing their development into blastocysts. Emphasizing the crucial role of correctly establishing active enhancers marked by H3K27ac modification in embryonic differentiation. Multi-omics data demonstrates high repeatability and quality. PCA clustering analysis reveals a clear differentiation path in the distribution of H3K27ac-modified histones, chromatin accessibility sites, and the transcriptome from the 8-cell stage to the blastocyst. Differences in H3K27ac histone modification distribution between the ICM and TE indicate the presence of tissue-specific active enhancers in blastocyst. The disparities in H3K27ac modification distribution between blastocyst ICM and TE primarily occur in distal gene regions rather than proximal ones. Moreover, these distal enhancers exhibit a positive correlation with the expression of lineage-specific genes, suggesting a probable regulatory role of distal enhancers in gene modulation during embryonic differentiation. Tissue-specific enhancer-promoter (EP) interaction loops emerge during embryonic differentiation, allowing direct interaction between tissue-specific active enhancers and promoters, thereby activating the expression of tissue-specific genes. Limitations, reasons for caution While DNA sequences with H3K27ac modification are currently considered markers for active enhancers, H3K4me modification is another histone marker for enhancers. However, enhancers associated with H3K4me modification may not necessarily be in an activated state. Future research can simultaneously incorporate appropriate detection methods for H3K4me modification. Wider implications of the findings After elucidating the mechanisms through which active enhancers regulate mouse differentiation, comparing the differences in active enhancers during the first lineage differentiation process between mice and humans can help identify factors that enhance human blastocyst quality. This can potentially improve clinical outcomes in assisted reproduction. Trial registration number not applicable

P-076 Role of N-DRC complex in male fertility

Abstract Study question There are many causes of human infertility, what’s the role of Nexin-dynein regulatory complex (N-DRC) in male fertility? Summary answer Many gene mutants in N-DRC components have been identified in infertile men, which affect the assembly of N-DRC complex and cause defects of sperm. What is known already human infertility affects 10–15% of couples, asthenozoospermia accounting for 18% of infertile men and is a common male infertility phenotype. Nexin-dynein regulatory complex (N-DRC) is a large protein complex regulating device in the flagellum and connects adjacent doublets of microtubules (DMTs). Some N-DRC components were reported to be associated with human disease, such as Primary ciliary dyskinesia (PCD). In recent years, more and more studies reported mutants in subunits of N-DRC are related to male infertility. For example, DRC1, DRC5 and DRC3 mutant were identified in patients displayed multiple morphological abnormalities of the flagella (MMAF) and obstructive azoospermia. Study design, size, duration We searched articles about N-DRC and male infertility in pumped and select research studies and the tidied up the gene mutants, gene knock out mouse modals and IVF outcomes of different N-DRC components in infertile men. Besides, we summarized the parameters of semen and ultrastructure of sperm flagella in these patients. Participants/materials, setting, methods We searched articles about N-DRC and male infertility in pumped and select research studies and the tidied up the gene mutants, gene knock out mouse modals and IVF outcomes of different N-DRC components in infertile men. Main results and the role of chance N-DRC is a highly conserved structural complex located in the axoneme of the flagellum and cilium, containing at least 11 different subunits (DRC1∼DRC11). Patients who were diagnosed as MMAF and male infertility carrying homozygous DRC1 mutants. TEM analyses of the patients’ sperm revealed the flagellar axoneme structure is disordered. A frameshift mutant of DRC5 was identified in an infertile male with asthenozoospermia, but the patient’s problem of infertility could be successfully solved by vitro Fertilization (IVF). A recent study identified two bi-allelic DRC3 frameshift variants in two unrelated patients, one patient displayed MMAF and the other present obstructive azoospermia. The sperm flagella function of DRC5 deletion mice was abrogated but flagellum ultrastructure remained normal and the mice behaved as asthenozoospermia and sterile. DRC3, DRC7 and DRC9 knockout mice exhibit MMAF and male infertility, and in DRC7 KO mice, the axoneme of sperm is disorganized and it was difficult to detect the ‘9 + 2’ microtubule structure. DRC1 mutant mice on the C57BL/6 background died before puberty, KO DRC6 in mice neither has no obvious effects on sperm motility function nor destroy the “9 + 2” arrangement of microtubules in cilia, and male mice was fertile. Limitations, reasons for caution More samples of MMAF and asthenozoospermia men to sequence N-DRC gene, and track their assisted reproductive outcomes. Wider implications of the findings The conclusions in our study uncover certain mechanism of N-DRC deficient in MMAF and asthenozoospermia phenotype, which could provide a new basis for diagnosis of male infertility. Trial registration number not applicable

P-727 The impact of antidepressants and anxiolytics on assisted reproductive technology treatment outcomes

Abstract Study question Does antidepressant and anxiolytic use impact the chance of live birth after assisted reproductive technology (ART) treatment? Summary answer For women using antidepressants, ART treatment is as successful as for others. However, concomitant anxiolytic use was associated with failure to succeed. What is known already Infertility and depression are prevalent disorders in women of reproductive age and may occur coincidently. Antidepressants constitute the mainstay in treatment for moderate-severe depression, and selective serotonin reuptake inhibitors (SSRIs) are among the most commonly used antidepressants because they, in general, cause few side effects. However, antidepressant use in women undergoing ART treatment may be considered to balance the possible risks of untreated depression against fetal drug exposure and the efficacy of ART treatment. Furthermore, up to 50% of patients with depression also meet the criteria for anxiety disorders and may be treated with anxiolytics. Study design, size, duration This is an observational cohort study based on the Danish health registries. The use of antidepressants, i.e. as current or recent users, and anxiolytics or hypnotics at embryo transfer was obtained from the prescription register and information on all ART treatments and childbirths between 2006 and 2019 from the Danish ART register and the Danish birth register. Participants/materials, setting, methods The study population included 130,170 embryo transfers in 48,156 women undergoing ART treatment. A total of 1505 women were exposed to antidepressants at the time of embryo transfer, and 1160 women were recent users who discontinued before embryo transfer. We used mixed-effects logistic regression analyses. The association between antidepressants and anxiolytics respectively and ART was analyzed taking potential confounders into account estimating the odds ratios(OR) of either a biochemical pregnancy, clinical pregnancy, or live birth. Main results and the role of chance The adjusted OR(aOR) and corresponding 95% confidence interval (CI) of a biochemical-, and clinical pregnancy in women with current use of antidepressants at time of embryo transfer were 0.87 (95% CI: 0.73-1.04) and 0.84 (95% CI: 0.57-1.20), respectively, and 0.90 (95% CI: 0.67-1.19) for live birth. For women who discontinued antidepressant use before embryo transfer (labeled recent users), the aORs for a biochemical-, clinical pregnancy and live birth were 0.95 (95% CI: 0.76-1.17), 0.71 (95% CI: 0.46-1.09) and 1.02 (95% CI: 0.72-1.45), respectively. When analyzing women who use or recently used antidepressants in combination with anxiolytics or hypnotics the chance of a live birth was decreased markedly with an aOR of 0.15 (95% CI: 0.04-0.51). Based on nationwide data from more than a decade of ART treatments, we found that the use of antidepressants at the time of embryo transfer was not associated with a decreased chance of biochemical or clinical pregnancy, or live birth. However, when women treated with antidepressants also used anxiolytics or hypnotics, the chance of a live birth was significantly decreased though based on a limited number of patients suggesting a point of further investigation and attention. Limitations, reasons for caution As in all observational studies, it can never be ensured that the patients take the medicine, but we used information on prescriptions for antidepressants/anxiolytics from the prescription register using at least two redeemed prescriptions within 182 days before, and until 182 days after the date of embryo transfer. Wider implications of the findings Use of antidepressants at the time of embryo transfer was not associated with a decreased chance of biochemical or clinical pregnancy, or live birth. However, when antidepressants are used concomitantly with anxiolytics or hypnotics, the chance of live birth is significantly decreased suggesting the need for further investigation and attention. Trial registration number The study was funded by The Region of Southern Denmark and Merck.

P-248 Egg vitrification related to female age: survival and laboratory outcome of 1233 vitrified-thawed oocytes in non-medical autologous use

Abstract Study question Is the female age at retrieval a determinant factor for the viability and potential of the thawed eggs? Summary answer In patients over 40, thawed oocytes display more abnormal fertilization, develop less in extended culture and yield less top quality blastocyst. What is known already Oocyte cryopreservation is a well-established procedure especially since the advent of the vitrification technique. The successful outcomes achieved with vitrification encourage its application in fertility preservation (FP) for medical and non-medical reasons. Besides, the age-related decline in women’s fertility and the oocyte quality negatively affect the outcome of assisted reproduction. Some studies have reported that age at oocyte retrieval strongly influences reproductive prognosis after FP. However, the appropriate age for non-medical FP is still debated. Although oocyte freezing at a younger age is recommended, there should be a balance between a favourable outcome and the cost-effectiveness of the strategy. Study design, size, duration Retrospective study on 166 patients with frozen-thawed oocytes who underwent ICSI during 2021-2023 in a single hospital. Oocyte vitrification had different purposes: banking for poor responders, to limit the number of embryos, and to solve unexpected situations where partner’s semen sample was unavailable. Patients were divided into three groups according to their age at retrieval (Group A ≤ 37 n = 77, Group B 37-40 n = 44, Group C ≥ 40 n = 45). Post-warming survival, fertilization, and embryological data were studied. Participants/materials, setting, methods 1233 oocytes were thawed. The intact oocytes were microinjected 1-2 hours after warming. Embryos were cultured in Sage 1-StepTM to blastocyst stage, and rated on daily basis. Embryo transfer was performed on day 3 or day 5 and remaining blastocysts were frozen on Day 5 or Day 6. Post warming survival, lysis after ICSI, fertilization, embryo development, blastulation rate (BR) and top blastulation rate (TBR), and morphokinetic features were studied in the 3 groups. Main results and the role of chance 84.6% of the 1233 oocytes (vitrified in a high-security closed system) survived the warming process. 1043 eggs were injected and the post-ICSI lysis rate was 11.2%. No significant differences were seen in thawing survival rate in groups A, B and C (85.6% vs. 83.5% vs. 83.8%, p=0.635) nor in the post-ICSI lysis rate (13.1% vs. 7.4% vs. 11.2%, p=0.072). 2PN fertilization rate was similar between the groups (67.8% vs. 61.3% vs. 67.7%, p=0.178) as well as the total number of embryos (303 vs 146 vs 188 p=0.605). However, triploid fertilization rate (3PN) statistically grew with age (2.8% vs. 5.8% vs. 6.5%, p=0.034). Significant differences were also reported in the global abnormal fertilisation rate (1PN+3PN) (6.4% vs 10.3% vs. 13.3%, p= 0.005). BR and TBR were statistically lower after the age of 40 (BR: 61.5% vs. 62.7% vs 40% p&amp;lt;0.001, TBR: 43.3% vs. 47.9% vs. 24.3% p&amp;lt;0.001). Morphokinetic features in the 3 groups were studied (abnormal cleavage pattern, tSB and tB). Overall results were similar between group A and group B but oocyte potential decreases drastically from the age of 40. Limitations, reasons for caution The major limitation of the study is its retrospective nature and the relative small size of each group. As most of patients were transferred during 2023, the pregnancies have not delivered yet so the live birth rate is not mentioned. Wider implications of the findings Studies have reported the impact of age at oocyte retrieval in case of medical FP in cancer and endometriosis. Here we report a negative effect on outcome after the age of 40 in non-medical FP. Patients should be informed that FP is not a certification of a successful outcome. Trial registration number not applicable

Smooth endoplasmic reticulum aggregates in oocytes associated with increased risk of neonatal birth defects: Ameta-analysis.

Previous studies have indicated the association between smooth endoplasmic reticulum aggregates (SERa+) and poorer medically assisted reproduction outcomes. However, the link between SERa+ and neonatal outcomes remains controversial and open for debate. A comprehensive meta-analysis on the relation between SERa+ and the risk of birth defects is needed. The literature search was conducted using the following databases: PubMed, Embase, Cochrane Libraries, Web of Science, and Chinese databases including China National Knowledge Infrastructure (CNKI) and Wan Fang from inception until July 2023. Risk ratio (RR) and 95% confidence interval (CI) were calculated by a fixed-effected model, while heterogeneity was assessed by forest plots and I2 statistic. Funnel plot was produced to assess publication bias. This meta-analysis has been registered on PROSPERO (CRD42022313387). The search resulted in 122 studies, 14 of which met the inclusion criteria. The analysis of birth defects revealed a higher risk (RR = 2.17, 95%CI 1.24 to 3.81, p = 0.007) in children derived from SERa+ cycle compared to SERa- cycles (711 vs. 4633). Meanwhile, in a subgroup analysis, the risk of birth defects was significantly increased in the SERa+ oocytes group as compared with the sibling SERa- oocytes group (RR = 3.53, 95%CI 1.21 to 10.24, p = 0.02). To conclude, our analysis indicated that SERa+ cycles/oocytes may have a potential risk of increased additional major birth defects comparing with SERa- cycles/oocytes. This conclusion may provide evidence-based support for clinicians in IVF clinical guidance and embryologists in prudent embryo selection strategy.

P-582 Analysis of assisted reproductive outcomes for gynecologic cancer survivors: a retrospective study

Abstract Study question Do women diagnosed with gynecologic cancer have worse pregnancy and neonatal outcomes of assisted reproductive technology (ART) ? Summary answer Decreased live birth rate was observed in female gynecologic cancer patients undergoing ART, particularly in endometrial cancers and cervical cancers. What is known already Gynecologic malignancies comprise approximately 13% of female cancer diagnoses, and oncological treatments such as surgery, chemotherapy, abdominal or pelvic radiotherapy, and hormonotherapy may induce infertility, premature ovarian failure, or early menopause, resulting in a loss of reproductive potential. ART has emerged as an important option for cancer patients undergoing treatment, but the reproductive outcomes of ART among cancer survivors remain poorly understood. Limited research exists regarding the impact of a history of gynecologic cancer on reproductive outcomes of ART. Study design, size, duration 136 women diagnosed with gynecologic cancer who underwent their first IVF/ICSI treatment between 2013 and 2021 were retrospectively enrolled in this study. 241 infertile women without any history of cancer were matched to the cancer group. Participants/materials, setting, methods Women diagnosed with gynecologic cancer who underwent their first IVF/ICSI treatment between 2013 and 2021 were included. Infertile women without any history of cancer were matched to the cancer group. The primary outcome was the cumulative live birth rate. A propensity score-based patient-matching approach was adopted to ensure comparability between individuals with and without specific cancer type. Main results and the role of chance A total of 136 patients with a history of gynecologic cancer and 241 healthy infertile controls were included in this study. Endometrial cancer constituted 50.70% of the cases and cervical cancer constituted 34.60% of the cases. The cancer group exhibited significant shorter duration of stimulation, lower levels of estradiol, lower number of retrieved oocytes, day-3 embryos, and blastocysts compared to the control group (P &amp;lt; 0.05). The cumulative live birth rate of the gynecologic cancer group was significantly lower than that of the control group (36.10% vs. 60.50%, P &amp;lt; 0.001). Maternal and neonatal complications did not significantly differ between the groups (P &amp;gt; 0.05). The endometrial cancer and cervical cancer groups showed significantly lower cumulative live birth rates than their matched controls (38.60% vs. 64.50%, P = 0.011 and 24.20% vs. 68.60%, P &amp;lt; 0.001, respectively). Limitations, reasons for caution Our study was limited by the retrospective design in nature and small sample size of its patient cohort. In addition, the effect of cancer treatments was not evaluated specifically in the current study due to the limited number of patients and possible inconsistency in surgical operations. Wider implications of the findings Our study provides data support for further exploration of the effects of endometrial cancer, cervical cancer, and other gynecologic cancers on fertility. Comprehensive ovarian function assessment, detailed counseling, individualized fertility guidance, and close follow-up after ART therapy are recommended for patients with gynecologic malignancy history. Trial registration number Not applicable

O-019 Outcomes and correlations between traditional Intracytoplasmic Sperm Injection (ICSI) method and a new AI-enhanced system

Abstract Study question In the context of ICSI, how does the Sperm Tracer Deep Neural Network analysis compare to the embryologist’s sperm selection technique? Summary answer Sperm Tracer AI system significantly improves fertilization rates, offering a standardized, objective approach to sperm selection, but nuanced correlations highlight complexities in sperm parameters. What is known already Traditional sperm selection during ICSI relies on subjective, manual methods, introducing variability and affecting pregnancy success. In response, AI applications, aim to enhance objectivity, potentially improving pregnancy rates. Study design, size, duration Between September and December 2023, the study examined 26 fresh ICSI cycles in a total of 268 sibling oocytes. Men aged 18 years and above with 34.4 mean women’s age. The Sperm Tracer group underwent single embryo culture, while the Embryologists' group underwent group embryo culture. The study analyzes statistical differences in fertilization rates, day 3 and day 5 embryo development. The study duration aligns with the analyzed cycles. Participants/materials, setting, methods The study compared real-time AI-assisted sperm evaluation with the Sperm Tracer system to the traditional ICSI method by embryologists. Correlation analysis was performed for the following kinematic parameters: Amplitude of Lateral Head displacement (ALH), Dance (DNC), Linearity (LIN), Straightness (STR), Wobble (WOB), Curvilinear Velocity (VCL), Linear Velocity (VSL) and Average Path Velocity (VAP). Statistical analysis involved Odds Ratios (OR) with 95% confidence intervals, Mann-Whitney U tests and correlation analyses by Pearson’s coefficient. Main results and the role of chance The Sperm Tracer system exhibited a noteworthy enhancement in fertilization rates (OR: 0.46, 95% Confidence Intervals (CI) [0.23, 0.89], p = 0.0224) but no significant differences in cleavage stages, blastocyst formation, or the production of good-quality embryos compared to conventional methods. For correlation analysis used two groups of sperm population. Group A comprises spermatozoa subjected to ICSI with the Sperm Tracer system, resulting in unsuccessful oocyte fertilization and Group B consists of spermatozoa that achieved successful fertilization. In Group A, correlation analysis identified significant negative associations between DNC and linear parameters (LIN, STR, WOB) (p &amp;lt; 0.001), highlighting the clinical relevance in sperm motility. Group B displayed a distinct negative correlation between DNC and LIN (p = 0.03941), indicating potential effects on sperm linear velocity. LIN in Group A exhibited strong negative correlations with WOB (p = 0.0001), moderate negative correlations with STR (p = 0.0028), and VCL (p = 0.0773). Moreover, ALH analysis in Group A unveiled robust negative correlations with LIN, STR, and WOB, suggesting clinical significance in sperm motility. These intricate findings underscore the Sperm Tracer system's primary advantage in optimizing fertilization rates, offering valuable insights for refining sperm selection strategies in assisted reproduction. Limitations, reasons for caution The study faces limitations due to a small sample, impacting generalizability. Divergence in culture embryo strategies introduces bias, necessitating validation in larger studies. The system includes morphology analysis, but no data analysis has been conducted. Exploring the correlation between morphology, considering embryo culture variations, and motility could provide valuable insights. Wider implications of the findings This study highlights AI applications like Sperm Tracer, showcasing their potential to improve assisted reproduction outcomes. Recognizing AI's impact on IVF success is vital for adoption. Integrating AI into clinic protocols may boost IVF efficiency, requiring training for embryologists to ensure effective utilization. Trial registration number Not applicable

O-291 Sonographic Symphony: a systematic review and network meta-analysis of the use of 2D, 3D, 4D ultrasound’s influence on embryo transfer outcomes

Abstract Study question To comprehensively evaluate and compare pregnancy and live birth rates following clinical touch, 2D, 3D or 4D ultrasound guided embryo transfer of a blastocyst. Summary answer The use of 3D and 4D ultrasound is not superior to 2D transabdominal ultrasound, but all forms of ultrasound are superior to clinical touch alone. What is known already While several factors contribute to the overall success of embryo transfer, the accurate placement of the embryo within the uterine cavity remains a critical determinant of implantation and subsequent pregnancy. While two-dimensional (2D) ultrasound provides basic imaging during embryo transfer, assisting with catheter placement, three-dimensional (3D) ultrasound adds volume and spatial information, aiding in assessing the endometrial cavity and embryo positioning. Meanwhile, four-dimensional (4D) ultrasound introduces real-time visualisation, offering dynamic insights into embryo movement and potential implantation. Although 2D remains a standard for most transfers, there is a growing interest in leveraging advanced imaging technologies to improve assisted reproductive outcomes. Study design, size, duration We searched bibliographical databases, including the Cochrane Central Register of Controlled Trials, EMBASE and Medline, from inception until August 2023. Randomised controlled trials that compared 2D transabdominal (2D TAUS), 2D transvaginal (2D TVUS), 3D and 4D ultrasound guided embryo transfer with one another or with clinical touch were included. The primary outcome was live birth rate (LBR), and the secondary outcome was clinical pregnancy rate (CPR) per transfer. Participants/materials, setting, methods Two reviewers independently screened selected studies and extracted the data. Pairwise and network meta-analyses (NMA) were conducted according to the technique used. Effect estimates were odds ratio (OR) with 95% confidence interval (CI) for each outcome respectively. Summary estimates were calculated using random-effects methods and quality assessment was performed using GRADE. Main results and the role of chance Twenty-eight trials, reporting data from 10,521 embryo transfer procedures, were included. Four different ultrasound modalities were evaluated: 2D transabdominal guided embryo transfer (2D TAUS) (21 trials), 2D transvaginal guided embryo transfer (2D TVUS) (four trials), 3D transabdominal guided embryo transfer (one trial) and 4D transabdominal guided embryo transfer (one trial). Compared to 2D TAUS, there was no difference in LBR in the network meta-analysis when using 2D TVUS (OR 1.14; 95% CI: 0.75-1.74), 3D ultrasound (OR 0.98; 95% CI: 0.49-1.97) or 4D ultrasound modality (OR 1.28; 95% CI: 0.56- 2.94). The use of clinical touch was associated with lower LBR (OR 0.71; 95% CI: 0.49-1.03). Compared to 2D TAUS, there was no difference in clinical pregnancy rate in the network meta-analysis when using 2D TVUS (OR 1.17; 95% CI: 0.86-1.58), 3D ultrasound (OR 0.97; 95% CI: 0.57-1.63) or 4D ultrasound modality (OR 1.30; 95% CI: 0.71- 2.38). The use of clinical touch was associated with lower CPR (OR 0.74; 95% CI: 0.64-0.85). Limitations, reasons for caution Variability in study designs, methodologies, and participant characteristics across primary studies can complicate the synthesis of results. There was only one study directly comparing 3D and 4D interventions which resulted in sparse data, affecting the precision and reliability of estimates. Wider implications of the findings Published evidence supports the continued use of 2D transabdominal ultrasound guided embryo transfer in routine clinical practice. There is insufficient evidence to support the introduction of 3D and 4D ultrasound when performing embryo transfers. Larger randomised controlled trials specifically assessing the superiority of these modalities are recommended. Trial registration number not applicable

Imprinting disorders in children conceived with assisted reproductive technology in Sweden

ObjectiveTo assess whether the use of assisted reproductive technology for conception is associated with imprinting disorders in children and the impact of parental factors related to infertility. DesignA nationwide register-based cohort study. SubjectsAll liveborn singletons in Sweden (N = 2 084 127) between 1997-2017 with follow-up to December 31, 2018. ExposureThe use of specific methods implemented in the assisted reproductive technology Main Outcome MeasuresThe International Classification of Diseases version 10 was used to identify three distinct imprinting disorder groups: Prader-Willi/Silver-Russell syndrome, Beckwith-Wiedemann syndrome, and central precocious puberty. The Cox model combined with inverse probability treatment weights were used to estimate weighted hazard ratio (wHR) with 95% confidence interval (CI), accounting for multiple confounders. ResultsA total of 1044 children were diagnosed with the disorders of interest, and 52 of them were conceived with assisted reproductive technology. The overall risk of being diagnosed with any of the studied imprinting disorders was elevated in children conceived with ART compared to all other children (HR 1.84, 95% CI: 1.38-2.45). After adjusting for parental background factors, the association was partially attenuated (wHR 1.50, 95% CI: 0.97-2.32), but remained also in the weighted comparison restricted to children of couples with known infertility (wHR 1.52, 95% CI: 1.05-2.21). For the specific diagnoses of Prader-Willi/Silver-Russell syndrome and Beckwith-Wiedemann syndrome, compared to children of couples with known infertility, children conceived with assisted reproductive technology showed a small excess risk, which could not be distinguished from the null (wHR 1.56 [95% CI: 0.93-2.62] and 1.80 [95% CI: 0.99-3.28], respectively). Further subgroup analysis showed that the combined use of intra-cytoplasmic sperm injection and cryopreserved embryos was associated with higher risk of both Prader-Willi/Silver-Russell syndrome (wHR 4.60, 95% CI: 1.72-12.28) and Beckwith-Wiedemann syndrome (wHR 6.69, 95% CI: 2.09-21.45). The number of central precocious puberty cases in children conceived with assisted reproductive technology was too small (N=3) to make any meaningful inference. ConclusionThe combined use of intra-cytoplasmic sperm injection and cryopreserved embryos was associated with small elevated risks of Prader-Willi/Silver-Russell syndrome and Beckwith-Wiedemann syndrome in children, independent of parental factors related to infertlity.

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.

Leukocytospermia and/or Bacteriospermia: Impact on Male Infertility.

Infertility is a globally underestimated public health concern affecting almost 190 million people, i.e., about 17.5% of people during their lifetime, while the prevalence of male factor infertility is about 7%. Among numerous other causes, the prevalence of male genital tract infections reportedly ranges between 10% and 35%. Leukocytospermia is found in 30% of infertile men and up to 20% in fertile men. Bacterial infections cause an inflammatory response attracting leukocytes, which produce reactive oxygen species (ROS) and release cytokines, both of which can cause damage to sperm, rendering them dysfunctional. Although leukocytospermia and bacteriospermia are both clinical conditions that can negatively affect male fertility, there is still debate about their impact on assisted reproduction outcomes and management. According to World Health Organization (WHO) guidelines, leukocytes should be determined by means of the Endtz test or with monoclonal antibodies against CD15, CD68 or CD22. The cut-off value proposed by the WHO is 1 × 106 peroxidase-positive cells/mL. For bacteria, Gram staining and semen culture are regarded as the "gold standard", while modern techniques such as PCR and next-generation sequencing (NGS) are allowing clinicians to detect a wider range of pathogens. Whereas the WHO manual does not specify a specific value as a cut-off for bacterial contamination, several studies consider semen samples with more than 103 colony-forming units (cfu)/mL as bacteriospermic. The pathogenic mechanisms leading to sperm dysfunction include direct interaction of bacteria with the male germ cells, bacterial release of spermatotoxic substances, induction of pro-inflammatory cytokines and ROS, all of which lead to oxidative stress. Clinically, bacterial infections, including "silent" infections, are treatable, with antibiotics being the treatment of choice. Yet, non-steroidal antiphlogistics or antioxidants should also be considered to alleviate inflammatory lesions and improve semen quality. In an assisted reproduction set up, sperm separation techniques significantly reduce the bacterial load in the semen. Nonetheless, contamination of the semen sample with skin commensals should be prevented by applying relevant hygiene techniques. In patients where leukocytospermia is detected, the causes (e.g. infection, inflammation, varicocele, smoking, etc.) of the leukocyte infiltration have to be identified and addressed with antibiotics, anti-inflammatories or antioxidants in cases where high oxidative stress levels are detected. However, no specific strategy is available for the management of leukocytospermia. Therefore, the relationship between bacteriospermia and leukocytospermia as well as their specific impact on functional sperm parameters and reproductive outcome variables such as fertilization or clinical pregnancy must be further investigated. The aim of this narrative review is to provide an update on the current knowledge on leukocytospermia and bacteriospermia and their impact on male fertility.

Effect of coenzyme Q10 supplementation on post-vitrification mouse embryo development

Objective: To investigate the effects of coenzyme Q10 (CoQ10) supplementation on post-vitrification embryo development and gross morphology. Methods: Balb/c mouse embryos were cultured in potassium simplex optimised medium (KSOM) with varying CoQ10 concentrations [0 (control), 20, 40, and 60 μΜ]. The most effective CoQ10 concentration (40 μM) was selected for subsequent post-vitrification morphology study. Embryos were randomly divided into four groups: Group A (non-vitrified without CoQ10), Group B (non-vitrified with CoQ10), Group C (vitrified without CoQ10), and Group D (vitrified with CoQ10), followed by vitrification at the 8-cell stage. Survival rates and development until the blastocyst stage were evaluated through morphological examinations using ASEBIR's system, distinguishing normal and abnormal embryos. Results: Supplementation of 40 μM CoQ10 significantly increased blastocyst formation (95%) compared to the control group (92%), 20 μM (62%), and 60 μM (56%) (P&lt;0.001). Following vitrification, Group D exhibited a significant increase in blastocyst formation (92%) compared to Group C (82%) (P&lt;0.05). Morphological assessments indicated superior embryo quality in Group B over Group D during the cleavage stage, morula, and blastocyst (P&lt;0.05). Conclusions: CoQ10 supplementation exhibits promising potential to enhance preimplantation embryo development, increase blastocyst formation rates, and improve embryo quality post-vitrification. This offers a promising approach to mitigate oxidative stress on embryos, potentially improving overall assisted reproductive technology outcomes.

Early GnRH-agonist therapy does not negatively impact the endometrial repair process or live birth rate.

To investigate whether different timings of GnRH-a downregulation affected assisted reproductive outcomes in infertile women with moderate-to-severe intrauterine adhesions (IUAs) accompanied by adenomyosis. A retrospective case series. An assisted reproductive technology center. The study reviewed 123 infertile women with moderate-to-severe IUAs accompanied by adenomyosis undergoing their first frozen-thawed embryo transfer (FET) cycles between January 2019 and December 2021. The majority of patients had moderate IUA (n=116, 94.31%). The average Basal uterine volume was 73.58 ± 36.50 cm3. The mean interval from operation to the first downregulation was 21.07 ± 18.02 days (range, 1-79 days). The mean duration of hormone replacement therapy (HRT) was 16.93 ± 6.29 days. The average endometrial thickness on the day before transfer was 10.83 ± 1.75 mm. A total of 70 women achieved clinical pregnancy (56.91%). Perinatal outcomes included live birth (n=47, 67.14%), early miscarriage (n=18, 25.71%), and late miscarriage (n=5, 7.14%). The time interval between uterine operation and the first downregulation was not a significant variable affecting live birth. Maternal age was the only risk factor associated with live birth (OR:0.89; 95% CI: 0.79-0.99, P=0.041). The earlier initiation of GnRH-a to suppress adenomyosis prior to endometrial preparation for frozen embryo transfer did not negatively impact repair of the endometrium after resection.

Diverse impacts of female chromosomal polymorphisms on assisted reproduction outcomes: a retrospective cohort study

BackgroundThe effects of female chromosomal polymorphisms (FCPs) on various aspects of reproductive health have been investigated, yet the findings are frequently inconsistent. This study aims to clarify the role of FCPs on the outcomes of in vitro fertilization (IVF) and intracytoplasmic sperm injection (ICSI).MethodsThis retrospective cohort study comprised 951 couples with FCPs and 10,788 couples with normal karyotypes who underwent IVF/ICSI treatment at Peking University Third Hospital between 2015 and 2021. The exposure was FCPs. The embryological outcomes and clinical outcomes were compared.ResultsThe FCPs, as a whole, compromised the oocyte maturation rate (76.0% vs. 78.8%, P = 0.008), while they did not adversely affect other IVF/ICSI outcomes. Further detailed analyses showed that every type of FCPs contributed to the lower oocyte maturation rate, particularly the rare FCPs (69.0% vs. 78.8%, P = 0.008). The female qh + was associated with a higher normal fertilization rate (63.0% vs. 59.2%, adjusted P = 0.022), a higher clinical pregnancy rate (37.0% vs. 30.7%, adjusted P = 0.048), and a higher live birth rate (27.0% vs.19.0%, adjusted P = 0.003) in couples undergoing IVF. Conversely, in couples undergoing ICSI, female qh + was found to be related to a lower normal fertilization rate (58.8% vs. 63.8%, P = 0.032), a comparable clinical pregnancy rate (25.7% vs. 30.9%, P = 0.289), and a comparable live birth rate (19.8% vs. 19.2%, P = 0.880) compared to the control group. Additionally, an increased risk of preterm birth was observed in women undergoing IVF with multiple polymorphisms (62.5% vs. 16.9%, adjusted P < 0.001) and in women undergoing ICSI with pstk+ (36.4% vs. 15.4%, P = 0.036).ConclusionsOur research unravels the diverse impacts of various FCPs on IVF/ICSI outcomes, highlighting the detrimental effects of FCPs on oocyte maturation and the risk of preterm birth.

The association between triglyceride glucose-body Mass Index and in vitro fertilization outcomes in women with polycystic ovary syndrome: a cohort study

BackgroundPolycystic Ovary Syndrome (PCOS) is a common reproductive disorder that frequently affects fertility. The TyG-BMI (Triglyceride glucose-body mass) index is a newly explored parameter that may be linked to reproductive results in individuals with PCOS. Nevertheless, its connection with outcomes in In Vitro Fertilization (IVF) procedures remains uncertain.MethodsThis study included a total of 966 females who underwent IVF treatments for PCOS. At the baseline, the participants were categorized into four groups according to the quartiles of TyG-BMI measured prior to oocyte retrieval. Subsequently, the study compared the differences in clinical and laboratory outcomes among these four groups.ResultsPatients in higher TyG-BMI quartiles exhibited a decreased number of retrieved oocytes, 2PN embryos, and available/high-quality embryos (P < 0.05 for Q1-Q4). Additionally, the multivariable regression analysis revealed that individuals in the top quartile of TyG-BMI had a lower count of accessible embryos (β = -0.224, P = 0.257) and a decreased number of high-quality embryos (β = -0.352, P = 0.028) in comparison to those in the lowest quartile. Nevertheless, there were no notable variances detected in the rates of pregnancy or live births among these quartiles. Furthermore, a linear correlation was noted between the TyG-BMI index and the quantity of accessible embryos (P-non-linear = 0.6, P-overall < 0.001), along with high-quality embryos (P-nonlinear = 0.026, P-overall = 0.006). In contrast, there was no notable linear correlation found between the TyG-BMI index and the available embryo rate (P-nonlinear = 0.60, P-overall = 0.8).ConclusionsThe results of this research emphasize the notable correlation between TyG-BMI and IVF results in females diagnosed with PCOS. The interplay of insulin resistance and disorders of lipid metabolism may indeed play a pivotal role in influencing the assisted reproductive outcomes of patients with PCOS. Considering these findings, TyG-BMI proves to be a valuable indicator for exploring this potential association.

Reactive oxygen species and their role in the andrological factor of couple fertility.

Reactive oxygen species play a significant role in male fertility and infertility. They are essential for physiological processes, but when their concentration becomes excessive, they can be a cause of various sperm pathologies. Seminal leukocytes and pathologically abnormal sperm are the primary sources of oxygen radicals in ejaculate. They negatively affect sperm quality, including DNA fragmentation and sperm motility impairment. Addressing increased concentrations of reactive oxygen species involves various appropriate lifestyle modifications and measures, including the use of antioxidants, treatment of urogenital infections, management of varicocele, weight reduction, and others. In many cases, these interventions can lead to adjustments in the condition and improvement in sperm quality. Such improvements can subsequently lead to enhanced outcomes in assisted reproduction or even an increased likelihood of natural conception. In some instances, the need for donor sperm may be eliminated. However, a key factor is adhering to a sufficiently prolonged treatment, which requires patience on the part of both, the physician and the patient.

Disruption in CYLC1 leads to acrosome detachment, sperm head deformity, and male in/subfertility in humans and mice

The perinuclear theca (PT) is a dense cytoplasmic web encapsulating the sperm nucleus. The physiological roles of PT in sperm biology and the clinical relevance of variants of PT proteins to male infertility are still largely unknown. We reveal that cylicin-1, a major constituent of the PT, is vital for male fertility in both mice and humans. Loss of cylicin-1 in mice leads to a high incidence of malformed sperm heads with acrosome detachment from the nucleus. Cylicin-1 interacts with itself, several other PT proteins, the inner acrosomal membrane (IAM) protein SPACA1, and the nuclear envelope (NE) protein FAM209 to form an ‘IAM–cylicins–NE’ sandwich structure, anchoring the acrosome to the nucleus. WES (whole exome sequencing) of more than 500 Chinese infertile men with sperm head deformities was performed and a CYLC1 variant was identified in 19 patients. Cylc1-mutant mice carrying this variant also exhibited sperm acrosome/head deformities and reduced fertility, indicating that this CYLC1 variant most likely affects human male reproduction. Furthermore, the outcomes of assisted reproduction were reported for patients harbouring the CYLC1 variant. Our findings demonstrate a critical role of cylicin-1 in the sperm acrosome–nucleus connection and suggest CYLC1 variants as potential risk factors for human male fertility.

Disruption in CYLC1 leads to acrosome detachment, sperm head deformity, and male in/subfertility in humans and mice

The perinuclear theca (PT) is a dense cytoplasmic web encapsulating the sperm nucleus. The physiological roles of PT in sperm biology and the clinical relevance of variants of PT proteins to male infertility are still largely unknown. We reveal that cylicin-1, a major constituent of the PT, is vital for male fertility in both mice and humans. Loss of cylicin-1 in mice leads to a high incidence of malformed sperm heads with acrosome detachment from the nucleus. Cylicin-1 interacts with itself, several other PT proteins, the inner acrosomal membrane (IAM) protein SPACA1, and the nuclear envelope (NE) protein FAM209 to form an ‘IAM–cylicins–NE’ sandwich structure, anchoring the acrosome to the nucleus. WES (whole exome sequencing) of more than 500 Chinese infertile men with sperm head deformities was performed and a CYLC1 variant was identified in 19 patients. Cylc1-mutant mice carrying this variant also exhibited sperm acrosome/head deformities and reduced fertility, indicating that this CYLC1 variant most likely affects human male reproduction. Furthermore, the outcomes of assisted reproduction were reported for patients harbouring the CYLC1 variant. Our findings demonstrate a critical role of cylicin-1 in the sperm acrosome–nucleus connection and suggest CYLC1 variants as potential risk factors for human male fertility.