bluuebunny/crossref_metadata_2025 huggingface dataset:
O-023 Does flushing with oil-based contrast during hysterosalpingography (HSG) increase the live birth rate as compared to water-based contrast: a prospective cohort study
Authors: Lu M , Yan L , Zhang J
Published: July 2024

Study question

What is the effectiveness of oil-based contrast during HSG compared to water-based contrast?

Summary answer

HSG with oil-based contrast increased the live birth rate as compared to water-based contrast.

What is known already

Tubal flushing has long been thought to increase live birth rate in infertile couples. A large randomised clinical trial (RCT) in The Netherlands confirmed the long-term hypothesis that HSG with oil-based contrast (Lipiodolâ) increased the live birth rate as compared to HSG with water-based contrast. We assessed whether this treatment effect was also achieved with oil-based contrast in Chinese women.

Study design, size, duration

We performed a prospective cohort study between May 4, 2020 and March 30, 2022. A total of 1187 women were included in the analysis.These women were categorized into two groups based on the contrast they used: the Oil-Soluble Contrast Media (OSCM) group, which employed poppy seed oil, consisting of 569 cases, and the Water-Soluble Contrast Media (WSCM) group, which used iodixanol, comprising 618 cases.

Participants/materials, setting, methods

This study was conducted at Shanghai Jiao Tong University International Peace Maternity and Child Health Hospital .The primary outcome was ongoing pregnancy rate within 6 months after the examination.Secondary outcomes included clinical pregnancies, miscarriage, ectopic pregnancy and thyroid Stimulating Hormone.

Main results and the role of chance

The ongoing pregnancy rate of the oil group is significantly higher than that of the water group (24.1% versus 17.2%, rate ratio [RR], 1.53; 95% confidence interval [CI], 1.14 to 2.06; P = 0.004). Women in the oil group also achieved higher clinical pregnancy rate (32.2% versus 22.2%, RR, 1.67; 95% CI, 1.27 to 2.18, P < 0.001), higher live birth rate (23.9% versus 16.8%, RR, 1.56; 95% CI, 1.16 to 2.09, P = 0.003), and bore higher risk of miscarriage (8.0% versus 5.0%, RR, 1.67; 95% CI, 1.02 to 2.72, P = 0.04) than water group.

Limitations, reasons for caution

The sample size along with the data was obtained from a cohort study in a single center.

Wider implications of the findings

Tubal flushing with oil-based contrast medium is an effective treatment to improve the pregnancy outcomes of infertile women that should be part of every infertility work-up.

Trial registration number

not applicable
O-062 New strategies in ovarian stimulation
Authors: Patki A
Published: July 2024

Controlled Ovarian stimulation (COH) has an objective to induce multiple ovulations in order to yield a multiple oocyte harvest and offer multiple embryos available for transfer thereby increasing the efficacy of ART.

The theory of a multicyclic development of follicles during the menstrual cycle has prompted new approaches to ovarian stimulation. Research has suggested that recruitable antral follicles are continuously present in the ovaries during the menstrual cycle, and ultrasonographic studies have demonstrated that multiple cohorts or ‘waves’ of 2–5 mm follicles are recruited continuously during a menstrual cycle. Because of the asynchrony between endometrial receptivity and embryo development with a ‘non-conventional start’ stimulation, all the embryos are generally cryopreserved and transferred subsequently. This deferred transfer policy is currently possible given the advances in vitrification techniques, with success rates comparable to those following transfer with ‘fresh’ embryos.

New stimulation approaches, together with advanced cryopreservation techniques, allow for a total ‘disarticulation’ between the time of the menstrual cycle, ovarian stimulation start and embryo transfer. This new approach to ovarian stimulation is particularly useful for women seeking fertility preservation, especially where a shortened time to starting cancer treatment is desirable. Also, poor responders could benefit from the new stimulation protocols by continuing ovarian stimulation after the first oocyte retrieval, thereby obtaining more oocytes or embryos compared with the conventional approach. In the quest to reduce the incidence of OHSS and make the clinic OHSS free various antagonist protocols are now routinely used.

These and other new protocols discussed will include Late initiation of stimulation, long acting antagonist. Individualised (iCOS) to include mild stimulation, random start, Dual stimulation and double trigger are done to improve efficacy and success rates in certain group of patients.

New drugs like Long acting FSH and oral antagonist with their results will be presented.

Pre-Treatment medications like Estrogens, Progestogens, Combined CoC, antagonist will also be discussed.
P-431 Perinatal outcomes after recurrent pregnancy loss: a systematic review and meta-analysis
Authors: Letourneau I , Zitour K , Candeliere J , Hick M , Wu C  Q
Published: July 2024

Study question

Are women with recurrent pregnancy loss (RPL) at higher risk of adverse perinatal outcomes in a subsequent pregnancy?

Summary answer

Women with a history of RPL have significant higher odds of adverse perinatal outcomes in a subsequent pregnancy.

What is known already

Studies have suggested that RPL women may have higher risks for obstetrical and perinatal complications such as hypertensive disorders, placental abruption, placenta accreta spectrum disorders, preterm delivery, and perinatal death in a subsequent pregnancy compared to those without an RPL history. However, the existing body of research presents a conflicting narrative, thus highlighting the need for a comprehensive understanding of the implications of RPL on pregnancy outcomes.

Study design, size, duration

A systematic review and meta-analysis was conducted. MEDLINE, EMBASE, Google Scholar and Cochrane databases were searched from inception until July 2023 for studies on RPL and adverse perinatal outcomes. Data was extracted by two independent reviewers.

Participants/materials, setting, methods

DerSimonian and Laird random effect meta-analyses were performed to pool effect estimates. Results were presented as odd ratios with their respective 95% confidence intervals. Subgroup analyses were conducted for those with unexplained RPL and by number of pregnancy losses. Sensitivity analysis using only high-quality studies and influence analysis were performed. The Newcastle-Ottawa Scale was used to assess for risk of bias.

Main results and the role of chance

The systematic search yielded 8,915 records, of which 42 studies (n = 5,619,124) were included in our meta-analysis. Women with RPL had higher odds of pre-eclampsia (OR 1.19; 95% CI, 1.03-1.37), pregnancy-induced hypertension (OR 1.24; 95% CI, 1.03-1.48), gestational diabetes (OR 1.76; 95% CI, 1.28-2.44), cesarean delivery (OR 1.65, 95% CI, 1.47-1.85), placental abruption (OR 1.57; 95% CI, 1.37-1.81), placenta previa (OR 1.93; 95% CI, 1.59-2.35; 7 studies), placenta accreta (OR 4.42; 95% CI, 3.09-6.33), preterm birth (OR 1.79; 95% CI, 1.65-1.93), very preterm birth (OR 2.58; 95% CI, 2.08-3.18), small for gestational age (OR 1.31; 95% CI, 1.15-1.78), congenital anomalies (OR 1.25; 95% CI, 1.02-1.53), stillbirth (OR 1.25; 95% CI, 1.04-1.51) and perinatal death (OR 2.04; 95% CI, 1.58-2.71) compared to women without a history of RPL. There was no association with aneuploidy.

Limitations, reasons for caution

Significant inter-study heterogeneity and inconsistent adjustment for confounders was noted across included studies.

Wider implications of the findings

RPL patients are at higher odds of adverse perinatal outcomes in a subsequent pregnancy, and therefore require additional counselling and monitoring in said pregnancy. There is an urgent need for further prospective research clarifying the relationship between RPL and perinatal outcomes.

Trial registration number

N/A
P-746 Recruitment in randomised controlled trials: a conundrum
Authors: Rikken J , Casteleijn R , Van der Weide M , Duijnhoven R , Goddijn M , Mol B  W , Van der veen F , Van Wely M
Published: July 2024

Study question

What factors are associated with recruitment failure within the pre-planned study period?

Summary answer

Trials with a no-treatment arm, over four inclusion criteria, without a previous pilot and with low funding, are more likely to experience recruitment failure.

What is known already

Randomised controlled trials are considered to be the best strategy in evaluating the effectiveness of medical interventions, but they are often hampered by failure to recruit enough patients with the pre-planned study period. This represents a form of research waste. Little is known on variables related to recruitment failure within the pre-planned study period.

Study design, size, duration

This study was designed as a nationwide cohort study and included 71 randomized controlled trials supported by the Dutch Consortium for Women’s Health Research. We included trials initiated between March 1st 2003 and March 1st 2022. We extracted study data and information on predefined barriers and facilitators.

Participants/materials, setting, methods

Primary outcome was recruitment target not achieved within 6 months after the pre-planned recruitment period. Secondary outcomes included recruitment target not achieved within an extension of at least 12 months and premature termination of the trial. We used logistic regression to evaluate the association between potential indicators of recruitment failure and expressed these as odds ratios (OR) with corresponding 95% confidence intervals (CI).

Main results and the role of chance

Of the 71 trials included in our cohort, recruitment was not achieved within the pre-planned study period with a maximal extension period of 6 months or 12 months in 38 (54%) and 33 trials (46%), respectively. Indicators associated with higher chances on recruitment failure were a no treatment arm (OR 8.53 (95% CI 1.01 to 72.36)), a compensation fee of less than 200 euros per included patient (OR 3.16 (95% CI 1.18 to 8.45)) and a funding of less than 350.000 euros (OR 4.30 (95% CI 1.56 to 11.64)). Indicators associated with lower chances on recruitment failure were less than 4 inclusion criteria (OR 0.31 (95% CI 0.10 to 0.97)) and a preceding pilot study (OR 0.53 (95% CI 0.15 to 1.85)).

Limitations, reasons for caution

If we could have accessed an even larger cohort of studies we might have been able to identify more potential indicators for recruitment failure.

Wider implications of the findings

Investigators and grantproviders should consider our findings before the start of their studies, to optimize recruitmentcircumstances. If a trial is destined to have a suspected long recruitment, it seems wise to consider the relevance and urgency of the research question, and then accept a longer recruitment period with its consequences.

Trial registration number

not applicable
P-778 Exploration of maternal proteome over pregnancy based on corpus luteum number at conception
Authors: Von Versen-Höynck F , Ramachandran D , Avdulahu F , Dewender R  T , Froböse W , Richter K , Schröder-Heurich B , Baker V  L , Winn V  D , Pich A
Published: July 2024

Study question

Does the maternal plasma proteome profile differ depending on corpus luteum (CL) number at conception?

Summary answer

The proteome profiles show differences depending on the number of CL at conception and some differentially regulated proteins may be linked to preeclampsia.

What is known already

Frozen embryo transfer (FET) in a programmed cycle is associated with a higher risk for preeclampsia. These women also experience impaired maternal vascular health in early pregnancy and differences in the endocrine environment, such as a lack of relaxin. These changes may be related to the absence of products from the CL.

Study design, size, duration

Prospective cohort study (N = 878) between June 2015 and December 2018 in an academic setting.

Participants/materials, setting, methods

Women with viable pregnancies were recruited at 6-8 weeks and compared by number of CL: no CL (programmed cycle FET), 1 CL (spontaneous pregnancy after infertility, modified natural cycle FET), and multiple CL (fresh IVF). Peripheral blood was collected, and plasma frozen at -80 °C. MS-based proteomics was used to determine proteome profiles of 4 participants in each group at 11-14 and 35-37 weeks’ gestation and 6-12 weeks postpartum.

Main results and the role of chance

Over 1000 protein groups were identified and 650 could be quantified in all samples. We found significant differences based on number of CL. Many of these proteins have been reportedly linked to preeclampsia. The results in the first trimester indicate the importance of aminopeptidases involved in the breakdown of angiotensin III. In addition, extracellular matrix proteins were found to be differentially regulated. Also, at the end of the third trimester certain proteins were differentially regulated among the groups.

Limitations, reasons for caution

This was a pilot study and unable to determine if differences are related to selection bias or could exist already pre-pregnancy independent of CL number at conception.

Wider implications of the findings

The findings warrant confirmation in a larger patient cohort. The detected differences in the proteome might shed light upon phenomena observed in women during pregnancy that lack a CL and as well as molecular mechanisms underlying preeclampsia.

Trial registration number

not applicable
O-114 Instructions for assembly of early mammalian embryos
Authors: Plachta N
Published: July 2024

The Plachta lab has pioneered the use of live-imaging approaches to discover how the preimplantation embryo forms in real time. Our recent findings help to transform the field of preimplantation development by showing how diverse processes occurring at the level of transcription dynamics, cytoskeletal organization, and the action of mechanical forces are integrated in real time to ensure proper embryo development. We extend our studies from mouse to human embryos, which enabled us to discover differences in embryo patterning between species and some of the events causing developmental errors, such as aneuploidy within the intact-developing system.
O-115 Ovarian function in transmasculine people
Authors: Moravek M
Published: July 2024

Transgender and gender diverse patients are increasingly being seen in fertility clinics for fertility preservation and/or family building, yet very little is known about the effects of gender-affirming hormones on reproductive capacity, precluding data-driven counseling. Specific to transmasculine individuals, human studies reporting the effects of gender-affirming testosterone on ovarian function are all observational in nature, many with only short-term testosterone exposure. Animal studies contribute to our understanding of the effect of testosterone on reproductive physiology, but have yet to be confirmed in humans. This lecture will review what is and isn't known about reproductive outcomes after gender-affirming testosterone, including whether testosterone can be continued during ovarian stimulation, and make suggestions for improving the clinical experience for transmasculine individuals seeking fertility care. Areas for future study will also be discussed.
O-150 Identification and validation of RPL7 and RCCD1 as potential biomarkers associated with immune infiltration in patients with thin endometrium
Authors: Sun J , Ji M , Zhu Y , Bukulmez O , Yeung W S , Sun J , Teng X , Chen M
Published: July 2024

Study question

For patients with thin endometrium (TE), what potential genes could be utilized for exploring the underlying mechanisms and clinical therapy?

Summary answer

Ribosomal protein L7 (RPL7) and RCC1 domain containing 1 (RCCD1) were suggested as specific biomarkers for TE.

What is known already

TE is an important contributor to embryo implantation failure and female infertility. However, the pathogenesis and mechanism of TE are not clear, and consequently, there is a paucity of data on effective targeted treatments for patients with TE.

Study design, size, duration

Endometrial cells were collected from 18 women with TE and 18 normal controls for transcriptional sequencing. Experimental validation was based on other 10 endometrium samples, consisting of 6 TE samples and 4 controls.

Participants/materials, setting, methods

After performing transcriptional sequencing, differentially expressed genes (DEGs) were extracted between the two groups via bioinformatic analysis. Based on DEGs, we applied functional enrichment and LASSO analysis to identify potential biomarkers, whose expressions were then validated by quantitative real-time PCR and western blot. Immune infiltration as well as the correlation with immune cells was also explored in the study.

Main results and the role of chance

RPL7 and RCC1 were identified and validated to be potential biomarkers, which exhibited an overexpression in TE at both mRNA and protein levels. An abnormal immune environment was found in TE, and correlation analysis showed that the two candidate genes correlated with atypical regulation of immune cells.

Limitations, reasons for caution

It might not be adequate to evaluate TE by only considering the immunologic aspects. Moreover, prospective studies are required to determine whether RPL7 and RCC1 may be used for early clinical diagnosis and prediction of pregnancy outcomes.

Wider implications of the findings

Further understanding and better knowledge of the functions of RPL7 and RCCD1 can promote the development of targeted therapeutics to improve clinical outcomes of patients with TE.

Trial registration number

not applicable
O-173 Gastruloids: a pluripotent stem cell model of gastrulation and body plan engineering in development and disease
Authors: Martinez-Arias A
Published: July 2024

Gastrulation is the process whereby the mass of cells that results from fertilization is shaped into the body plan of an organism. Understanding this process in humans is important as the origin of several syndromes and birth defects, map to this period. However, addressing this period experimentally is difficult due to technical, and more importantly, obvious ethical constrains. Over the last few years, alternatives to study gastrulation have emerged from the realization of the self-organizing abilities of Pluripotent Stem Cells and led to a number of cellular models of the early stages of mammalian development. I will discuss these models, with a special focus on gastruloids, highlighting their strengths as well as their limitations.
O-180 On being whole – fantasy and the unthought known
Authors: Ramu T
Published: July 2024

Timo Ramu unexpectedly discovered, through a DNA test meant for a lighthearted exploration of ethnicity, that he was donor-conceived. From initially thinking he had no biological siblings, he found out he had 50 and counting. This revelation shocked his family and shattered his sense of identity. His parents, in trying to shield him, denied him knowledge of his true origin. He grapples with the implications of living in a "fantasy" crafted by others. The talk touches upon the importance of biological connections, the distinctions between biological and social relationships, and the profound impacts of withheld truths on trust and self-awareness.
O-191 Delayed fetal growth in pregnant women with polycystic ovary syndrome using serial ultrasonographic biometry
Authors: Wang F , Fu L , Yun F , Wang T , Zhou Y , Qu F
Published: July 2024

Study question

To investigate the effect of maternal polycystic ovary syndrome (PCOS) on intrauterine growth and perinatal outcome of offspring.

Summary answer

Maternal PCOS might have an adverse impact on fetal growth, represented by serial biometric measurements, and consequently lower the birth weight.

What is known already

Women with PCOS often have low fertility and high risk of certain pregnancy complications, including miscarriage, gestational diabetes mellitus (GDM), hypertensive disorders of pregnancy (HDP) and preterm birth (PTB). In addition, the offspring of PCOS mothers are susceptible to short- and long-term health consequences, such as metabolic diseases and neuropsychiatric disorders. And the suboptimal intrauterine environment caused by maternal PCOS condition might be the origin of impaired long-term health of the offspring.

Study design, size, duration

A total of 1153 pregnant women with pregestational diagnosis of PCOS and 1383 non-PCOS controls were included in the retrospective case-control study for clinical record and ultrasonographic database at women’s hospital, school of medicine, Zhejiang university over a 6-year period.

Participants/materials, setting, methods

Maternal demographic data, fetal biometric measurements with ultrasound, obstetrical and neonatal outcomes were analyzed. A meta-analysis of birth weight in PCOS offspring in the literature was performed to assess the relationship between the maternal PCOS and neonatal growth.

Main results and the role of chance

After adjustment for potential confounding variables, we found that biparietal diameter (BPD) and femur length (FL) were notably smaller in fetuses of women with PCOS than controls across the entire gestation. This effect still existed after excluding three major pregnancy complications of PCOS, including GDM, HDP and PTB. Although other fetal biometry including estimated fetal weight and birth weight in PCOS offspring were comparable with controls in our study, the meta-analysis of 22 studies showed that the birth weight in offspring of women with PCOS was lower than controls (WMD -37.813, 95%CL [-69.713, -5.913]), especially for offspring in South America (WMD -84.69, 95%CL [-166.69, -2.68]).

Limitations, reasons for caution

First, data of other fetal parameters like abdominal and head circumference was scanty. Second, neonatal anthropometric parameters apart from the birth weight were not included. Lastly, we did not have the long-term follow-up results of the offspring to explore the potential effect of delayed fetal biometry in their later life.

Wider implications of the findings

More attention needs to be paid to the smaller-than-average fetuses in PCOS mothers, and proper management may be needed during and after pregnancy. Further studies are needed to confirm the intrauterine growth pattern of the PCOS offspring and its clinical significance in the neonatal period and beyond.

Trial registration number

N/A
O-241 ESHRE guidance on embryo transfer: when to do DET?
Authors: Veleva Z , Alteri A , Arroyo G , Baccino G , Craciunas L , De Geyter C , Santos-Ribeiro S , Ebner T , Kordic K , Mertes H , Pavicic Baldani D , Rodriguez-Wallberg K , Rugescu I , Tilleman K , Woodward B
Published: July 2024

More than four decades have passed since the birth of the first infant conceived through the use of assisted reproductive technology (ART). During this time, ART has increased the chance for millions of people who have trouble conceiving to achieve a healthy pregnancy and birth. In the early years of IVF/ICSI, clinicians may have transferred all available embryos into the uterus because of the low overall treatment success at the time. However, with medical and technical improvements in ART, the transfer of multiple embryos led to a greater number of multiple pregnancies, compared to spontaneous pregnancies. Multiple pregnancies are associated with a wide range of maternal and neonatal complications. This is why at present, the transfer of one embryo is considered the preferable route towards the key objective of ART and at present this is recommended by several international and national professional organisations such as ESHRE and ASRM.

However, these recommendations are not followed in many countries, as can be observed in annual reviews. Double embryo transfer (DET) remains a viable alternative to elective single embryo transfer (eSET) in many clinics.

The aim of the presentation is to highlight the place of double embryo transfer in the current practice of ART while taking into account best evidence from clinical studies. The presentation will present results from ESHRE’s guideline on the number of embryos to transfer in IVF/ICSI as well as some results from the surveys of the guideline development group (GDG), the most important of which is the survey to European infertility patients performed together with Fertility Europe, Europe’s patient organisation.
O-244 AMH levels in infertile women in APAC region - prospective study in multiple countries, ethnicities and centres
Authors: Tzeng C  R
Published: July 2024

Secretary General, Asia Pacific Initiative on Reproductive (ASPIRE)

President, Taiwan Endometriosis Society

Founder & CEO, Taipei Fertility Center

AMH is produced by granulosa cells of small growing follicles in the ovary and serum AMH levels strongly correlate with the number of growing follicles, AMH has been used clinically as a marker for ovarian reserve. Despite being of similar ages, with similar AMH levels, durations of ovarian stimulation, and gonadotropin doses, Asian women have been documented as having decreased AFCs and lower yields of total and mature oocytes after ovarian stimulation compared to their Caucasian counter- parts. These earlier data suggested that the effect of ethnicity was as an independent marker of ovarian reserve and potentially subsequent reproductive outcomes.

We conducted the first multi-nation, multi-centre, and multi-ethnicity prospective cohort study from January 2020 to May 2021 to investigate serum AMH profiles among infertile Asian women. A total of 4613 women who visited fertility clinics at 12 IVF centres in 11 regions throughout Asia were recruited from Taiwan, China, Hong Kong SAR, India, Indonesia, Japan, Korea, Malaysia (2 centres), Thailand, Vietnam, and Singapore. The inclusion criteria were female partners aged 20–43years and naive to any ovarian stimulation cycle. The AMH measurement was performed using an AMH automated assay on a clinically validated platform.

Among the 12 centres that joined the study, most demonstrated racial homogeneity except for Singapore and Malaysia which presented a highly diverse ethnic profile, while others were supposedly monoethnic communities. Unexplained or male infertility is the most common infertility cause in most countries, except for China with a notably high incidence of tubal disease. Age, ethnicity, obesity, and PCOS are significantly associated with ovarian reserve. The highest rates of PCOS were found in Indians (18.6%), Malays (18.9%), and Vietnamese (17.7%), yet remarkably lean with lower obesity rate was found in Vietnamese. Serum AMH levels are not affected by smoking or GnRHa and OCPs pretreatment but are affected by endometrioma surgery. Moreover, the assay can be conveniently conducted on any day of the menstrual cycle and is not influenced by the use of fresh or frozen samples. The rate of decline in ovarian reserve increases with age in Asian women, at 0.13ng/ml/year at 25–33years old and 0.31ng/ ml/year at 33–43 years old. We also found that a cut-off at 4.0 ng/ml could be considered for PCOS diagnosis in the Asian region. Despite limitations, the results can help guide clinicians for personalized infertility treatment, and call for further investigation into the effects of genetic, environmental and lifestyle factors on ovarian reserve.
P-021 Development of recombinant Phospholipase C Zeta protein as a therapeutic intervention to overcome certain types of male infertility
Authors: Saleh A , Nomikos M
Published: July 2024

Study question

Developing recombinant Phospholipase C Zeta (PLCζ) protein using a bacterial expression system to be utilized for potential future replacement of Ca2+ ionophores during IVF treatments.

Summary answer

Our findings support the use of MBP-PLCζ recombinant protein as a therapeutic tool for treating certain types of male factor infertility associated with fertilization failure

What is known already

The long-term safety of calcium ionophores, which are exclusively used for treating cases of fertilization failure, is still questionable. PLCζ is an endogenous agent that may represent a better replacement for treating male infertility cases related to fertilization failure

Study design, size, duration

Maltose binding protein (MBP)-tagged PLCζ was produced by cloning wild-type PLCζ into a pET41MM vector and purified using affinity chromatography techniques. Circular dichroism (CD) was employed to evaluate the secondary structure and the thermal denaturation of PLCζ protein. Moreover, the PIP2 hydrolysis assay was used to test the enzymatic activity of PLCζ protein over time and under different storage conditions. Furthermore, the embryonic toxicity of the protein was tested using a chick embryo model.

Participants/materials, setting, methods

Several methods were used in this study including Molecular cloning, circular dichroism, and chick embryo model to investigate the embryonic viability.

Main results and the role of chance

Our findings demonstrated that PLCζ recombinant protein displayed a properly folded secondary structure mainly composed of alpha-helices. Moreover, the protein was effective in maintaining the enzymatic activity and Ca2+ sensitivity for 90 days when stored at -80oC. Moreover, exposing chicken eggs to MBP-PLCζ did not alter the embryonic viability when compared to the control group. Kaplan-Meier survival curve showed that the rate of survival did not differ between the treatment, control and sham groups until the endpoint of the study.

Limitations, reasons for caution

The toxicology assays used are not sufficient to confirm the safety of the protein. Further in vivo studies are required to investigate the effect of MBP-PLCζ protein on early embryonic development before being used as a replacement for calcium ionophores in cases of oocyte activation failure.

Wider implications of the findings

These findings can help develop a new therapeutic tool for treating cases of male infertility associated with oocyte activation failure.

Trial registration number

not applicable
P-109 Non-invasive Genetic Testing (niPGTA) and Laser Assisted Hatching (LAH) may improve pregnancy and ongoing pregnancy rates in patients of advanced maternal age
Authors: Banti M , Van Zyl E , Kafetzis D
Published: July 2024

Study question

Does niPGTA and LAH improve pregnancy and ongoing pregnancy rates in patients of advanced maternal age?

Summary answer

niPGTA increases pregnancy and clinical pregnancy rates in patients aged ≥38 years compared to those without tested embryos, while LAH may further improve pregnancy outcomes.

What is known already

Embryos release cell-free DNA (cf-DNA) in the IVF culture media, which can be analyzed for chromosomal constitution. The benefit that niPGTA holds over PGT-A is that it is a non-invasive test and could potentially produce comparable results. Therefore, the time to pregnancy and the risk of miscarriage could be reduced in comparison to an IVF treatment with non-tested embryos. Previous studies have shown high concordance with trophectoderm biopsies in PGT-A patients and improved outcomes with LAH in frozen-thawed cycles. However, there is a lack of studies investigating clinical pregnancy rates with niPGTA.

Study design, size, duration

Retrospective cohort study including 74 patients aged ≥38 years, who underwent Single Frozen Embryo Transfer (FET) from May 2021 to February 2023. Patients who had Endometrial Receptivity Assay (ERA) were excluded.

Participants/materials, setting, methods

32 couples had niPGTA; Spent Blastocyst Media (SBM) for each blastocyst were collected on Day 6 and analyzed using the EMBRACE test. Embryos were vitrified. 42 couples had their blastocysts vitrified on Day 5/6 without testing. LAH was introduced to all patients in October 2021. Before that, no LAH was performed. Embryos were transferred to patients 2 hours post-warming after LAH. Pregnancy test was performed 10 days after FET and clinical pregnancy was established.

Main results and the role of chance

There was a statistical significant increase of pregnancy rate (p = 0.017, Pearson’s Chi-Squared test) and a trend towards higher clinical pregnancy rate for niPGTA patients versus those who had a transfer of not tested embryos. Regarding LAH, there was a trend towards higher pregnancy and clinical pregnancy rates in both niPGTA and not tested group. Results for pregnancies and clinical pregnancies are presented in Table 1.

Limitations, reasons for caution

More patients are required to confirm clinical significance of pregnancy and clinical pregnancy rates of niPGTA over not tested embryos and if LAH can improve the outcomes. A review of patients who had previous implantation failures and miscarriages in addition to ERA could yield clearer conclusions.

Wider implications of the findings

The results of this study are encouraging for niPGTA combined with LAH for patients aged ≥38 years. Further studies on niPGTA and modifications of the protocol for increased concordance rates and shorter time in embryo culture should be explored.

Trial registration number

Not applicable
P-113 Comparison of mitochondrial function between expanded and hatching human blastocysts
Authors: Bayat H , Akbari M , Adib S , Movaghar B
Published: July 2024

Study question

Is there a difference in mitochondrial function between expanded and hatching fresh and vitrified blastocysts?

Summary answer

Freezing and thawing may lead to mitochondrial dysfunction.

What is known already

Mitochondria, as crucial cellular organelles, play role in energy production through the electron transport chain, cell motility and organelle movement. They also have a significant impact on embryonic development. Furthermore, during the process of embryonic hatching, the energy generated by these organelles can affect ion pump activity, blastocyst expansion, and collapse. Therefore, factors that stress embryonic mitochondria may contribute to embryogenesis and hatching. Cooling and warming, as stress factors, maybe result in organelle damage, protein denaturation, membrane disruption, and DNA fragmentation. Additionally, they can cause changes in mitochondrial function, leading to decreased ATP levels and increased ROS levels.

Study design, size, duration

In this study, vitrified and fresh human 8-cell embryos were collected from April 18, 2021, to July 19, 2023. The embryos were cultured until the blastocyst stage, and then four samples were assessed in four groups. These groups included vitrified expanded blastocysts, vitrified hatching blastocysts, fresh expanded blastocysts, and fresh hatching blastocysts.

Participants/materials, setting, methods

Vitrified embryos were sourced from the embryo bank of the Royan Institute, while fresh embryos were obtained from good-quality 3PN embryos or excess embryos from couples who did not wish to freeze them. A stock solution containing lipophilic cationic JC-1 dye in dimethyl sulfoxide was prepared. Blastocysts were then exposed to a concentration of 1 μg/ml of JC-1 diluted in HamsF10-HSA. After washing with HF-HSA, imaging was performed using a 2-photon confocal microscope.

Main results and the role of chance

Our results showed that mitochondrial membrane potential in expanded blastocysts of both fresh and vitrified eight-cell embryos was significantly higher than in hatching blastocysts of fresh and vitrified embryos (P < 0.05). However, there was no significant difference in mitochondrial function between the vitrified and fresh groups, neither in expanded nor hatching blastocysts.

Limitations, reasons for caution

Due to the human origin of the samples, the minimum necessary replicates were considered for each experimental group.

Wider implications of the findings

In this study, it appears that plump zona breakers can be detected in hatching blastocysts using confocal microscopy. Previous studies, such as the one conducted by Sathananthan, identified these cells using a TEM microscope.

Trial registration number

-
P-298 The use of propensity score matching to assess the effectiveness of the endometrial receptivity analysis in patients with recurrent implantation failure
Authors: Yu S , Zhang Y , Li N , Su Z , Li W , Lou H , Guan Y
Published: July 2024

Study question

The purpose of this study was to investigate the effect of ERA on pregnancy outcomes in patients with RIF.

Summary answer

The results of the study showed no significant difference in pregnancy outcomes in patients who received ERA compared to those who did not.

What is known already

One potential cause of implantation failure is abnormal endometrial receptivity, and how to objectively evaluate endometrial receptivity has been a matter of great concern. Endometrial receptivity analysis (ERA), a microarray-based test that assesses endometrial gene expression, may be valuable in predicting endometrial receptivity, but whether ERA improves pregnancy outcomes in patients with recurrent implantation failure (RIF) is currently controversial.

Study design, size, duration

We performed a retrospective cohort study analysis for a population of patients with RIF undergoing frozen embryo transfer (FET) cycles in the reproductive center of the Third Affiliated Hospital of Zhengzhou University from January 2019 to December 2022(n = 1598).

Participants/materials, setting, methods

FET cycles with personalized embryo transfer (PET) under ERA guidance were included in the ERA group (n = 43); after using propensity score matching (PSM), a total of 120 FET cycles were included as a control group. Pregnancy outcomes were compared between the two groups. Further, the relationship between the number of previous implant failures and the rate of implant window displacement was discussed. The factors affecting the window of implantation (WOI) displacement were also assessed.

Main results and the role of chance

There was no statistically significant difference in embryo implantation rate, clinical pregnancy rate, spontaneous abortion rate, and live birth rate between the ERA group and the matched control patients (P > 0.05).

Limitations, reasons for caution

Our study is a non-large sample size retrospective study, because ERA is not a routine test in assisted reproductive technology clinics.

Wider implications of the findings

In our study, PET did not significantly improve pregnancy outcomes in patients with RIF, no significant differences in pregnancy outcomes were found between patients with moderate and severe RIF after PET, and no underlying etiology affecting WOI displacement was identified.

Trial registration number

not applicable
P-427 Do endometrial leucocytes and angiogenesis in the peri-implantation period predict hypertensive disorders in the subsequent pregnancy?
Authors: Chen X
Published: July 2024

Study question

Do endometrial leucocytes and angiogenesis in the peri-implantation period from women with a history of recurrent reproductive failure predict hypertensive disorders in the subsequent pregnancy?

Summary answer

uNK cells, T cells, B cells and macrophages in the peri-implantation endometrium did not correlate with hypertensive disease of pregnancy in the subsequent pregnancy.

What is known already

Women with a history of recurrent reproductive failure are thought to have a higher risk of hypertensive disorders of pregnancy (HDP) subsequently. Immunological interactions and vascular factors in the placental bed are considered to be two important pathways for the initiation and development of HDP. However, the prognostic value of endometrial leucocytes and angiogenesis around the time of implantation in a cycle prior to conception and HDP in the subsequent pregnancy is still unclear.

Study design, size, duration

This is a prospective cohort study. A total of 694 women were included in this study from Jan 2019 to Jun 2023.

Participants/materials, setting, methods

Immunohistochemistry for vWF and multiplex immunofluorescence for endometrial leukocytes, including CD56+ uNK cells, CD3+ T cells, CD20+ B cells and CD68+ macrophages, were performed in the precisely timed LH + 7 endometrium. The subsequent pregnancy outcomes of the subjects were followed up as well.

Main results and the role of chance

Endometrial leukocytes including CD56+ uNK cells, CD3+ T cells, CD20+ B cells and CD68+ macrophages were found in the peri-implantation endometrium. There was no significant difference in the CD56+ uNK cells, CD3+ T cells, CD20+ B cells and CD68+ macrophages numbers percentage between women with and without gestational hypertension in the subsequent pregnancy. The median micro vessel density in women who developed gestational hypertension in the subsequent pregnancy (9.4 ± 3.1 /mm2 ) was significantly (P=0.038) lower than the MVD in those who did not (13.5 ± 4.9/mm2 ).

Limitations, reasons for caution

1. It is not clear whether the uNK cell number during the peri-implantation period is reflective of those in the first trimester when spiral artery remodelling occurs. 2. This study was carried out in women with reproductive failure and the results may not apply to the general populations.

Wider implications of the findings

Vascularization features in the endometrium may provide prognostic value of subsequent hypertensive disease of pregnancy.

Trial registration number

not applicable
P-571 Mutations in FOXO3 mutation cause human premature ovarian insufficiency
Authors: Sun L , Tong K , Huang G , Li J
Published: July 2024

Study question

Do variants in front fork transcription factor 3 (FOXO3) account for premature ovarian insufficiency (POI) in humans? Is there is a potential treatment for POI caused by FOXO3 mutations?

Summary answer

Heterozygous variants in FOXO3 cause human female infertility owing to POI by inducing oocyte apoptosis.

What is known already

Front fork transcription factor 3 (FOXO3) is a member of the forkhead box family of transcription factors. Previous study had reported that FOXO3 encodes an evolutionarily conserved primary regulator and an effective inhibitor of primordial follicle activation. The null mutants of Foxo3 or its ortholog in multiple organisms displayed POI owing to deficiencies in maintaining the ovarian reserve. However, whether FOXO3 variants can underline POI phenotype in humans were unclear, and the causal relationship has not yet been established with functional evidence.

Study design, size, duration

107 primary infertility families with POI were recruited for pathogenic variants screening. The proband with and her parents were verified for FOXO3 variants.

Participants/materials, setting, methods

All the patients were diagnosed with POI were confirmed by clinical analyses. Exome sequencing and subsequent bioinformatic analyses were performed to screen for candidate pathogenic variants. The pathogenicity of identified variants was assessed and studied in vivo in mice carrying the equivalent mutations.

Main results and the role of chance

Novel heterozygous variant in FOXO3 was identified in the patient with POI. Mice modeling the Foxo3 variant identified in patients recapitulated the ovarian reserve defects of patients, confirming the pathogenicity of the identified variants. The mutation caused constitutive activation of FOXO3 lead to POI by inducing oocyte apoptosis, which will facilitate the genetic diagnosis of POI in patients and provide a potential therapeutic target for female fertility.

Limitations, reasons for caution

A limitation of the current study is the small sample size. More primary infertility patients with POI should be screened to enrich the pathogenetic mutations of FOXO3.

Wider implications of the findings

FOXO3 was detected as a novel gene underlie human POI, thus providing new targets for the molecular diagnosis and treatment.

Trial registration number

the National Natural Science Foundation of China (NSFC) (Grant number:82302083)
P-598 Novel inactivating homozygous mutation of the luteinizing hormone/choriogonadotropin receptor (LHCGR) and first live birth after in vitro maturation (IVM): a breakthrough
Authors: Labrosse J , Sifer C , Cedrin I , Peigne M , Grynberg M
Published: July 2024

Study question

How can pregnancy and live birth be achieved in patients with inactivating homozygous luteinizing hormone/choriogonadotropin receptor (LHCGR) mutation?

Summary answer

We report the first live birth after IVM in a patient with a novel homozygous inactivating LHCGR mutation (exon 6,c.470A>G).

What is known already

LH plays a fundamental role in female reproductive physiology and is responsible for steroidogenesis, oocyte maturation, ovulation and subsequent progesterone production by the corpus luteum. LH binds to the LHCGR located on the membrane of theca cells and mature granulosa cells.

Inactivating mutations of the LHCG receptor lead to the impossibility to obtain final oocyte maturation both during natural cycles and after ovarian stimulation for in vitro fertilization purpose.

Until now, egg donation represented the only option to treat their infertility.

Study design, size, duration

A case report.

Participants/materials, setting, methods

A 35 year-old nulliparous patient was referred to our university hospital for primary infertility.

Main results and the role of chance

The 35-year-old nulliparous patient presented with primary spaniomenorrhea but timely and spontaneous onset of secondary sexual characteristics. Serum LH levels were high (ranging from 15 to 30 IU/L) and to a lesser extent so were FSH levels. The ovarian reserve was normal for age, as assessed by serum AMH levels and ultrasound. There was no argument for polycystic ovarian syndrome, 21-hydroxylase deficiency, Cushing’s syndrome or hyperprolactinemia.

Two previous attempts of controlled ovarian stimulation with gonadotropins and ovulation trigger with hCG and GnRH-agonist trigger had failed, resulting in low estradiol levels despite correct follicular growth on ultrasound and absence of ovulation after trigger.

We identified by genetic analysis a novel homozygous inactivating LHCGR mutation (exon 6, c.470A>G) that had never been described previously.

IVM was performed. A total of 7 oocytes were obtained after IVM, resulting in 4 Day 3 embryos. All embryos were frozen.

Subsequently, 2 Day 3 embryos were replaced after endometrial preparation by hormone replacement therapy.

The patient became pregnant and gave birth to a healthy baby. Two Day 3 embryos remain.

Limitations, reasons for caution

By definition, a case-report requires further studies to confirm our findings.

Wider implications of the findings

We describe a novel inactivating LHCGR mutation with subsequent live birth after IVM. As a growing number of gonadotropin receptor mutations are identified, this live birth successfully obtained places IVM as the only reliable option for these patients to conceive with their own eggs.

Trial registration number

not applicable