bluuebunny/crossref_metadata_2025 huggingface dataset:
O-142 SARS-CoV-2 vaccination and IVF cycle outcomes
Authors: Anis heusler A , Weintraub A , Stein D , Kulak D
Published: June 2022

Study question

Does recent mRNA SARS-CoV-2 vaccination effect IVF pregnancy rate?

Summary answer

We observed no influence of mRNA SARS-CoV-2 vaccine on patients’ IVF cycle outcomes; vaccination did not change pregnancy rates.

What is known already

Two recent studies showed that the BNT162b2 mRNA SARS-COV-2 vaccine does not appear to have negative effects on oocytes during controlled ovarian stimulation. Assessment of follicular fluid after vaccination (1) concluded that there was no affect on the follicle quality or function. An additional study of 36 patients that underwent IVF both immediately before and immediately after vaccination found no change in ovarian yield or embryo quality (2).

Study design, size, duration

A retrospective case control study including 385 women from January 1, 2021 to May 31st of the same year.

Participants/materials, setting, methods

The study group included patients that received 2 doses of the Pfizer SARS-COV-2 vaccine between 7-180 days prior to starting IVF and without a history of COVID infection. Controls included unvaccinated patients from the same time period and historical age matched patients from our hospital from the same months in prior years. 385 patients under the age of 40 years met the inclusion criteria, 349 in the control and 36 in the vaccinated group.

Main results and the role of chance

Demographic and clinical characteristics were comparable between groups. The pregnancy rate was 33.3% (n = 12) in the vaccinated group vs 31.8% (n = 111) in the control group (p > 0.1). There were no statistically significant differences between the control and vaccine groups with a small non-significant trend to higher pregnancy rates in vaccinated patients: pregnancy rate OR for vaccination group was 1.17 (95% CI 0.56-2.44). 

Limitations, reasons for caution

Our study size was small and was observational leaving it open to confounding and selection bias regarding who chose to receive the vaccine and those in high risk groups for COVID infection. Additionally, longterm effect of the vaccine and booster were not assessed.

Wider implications of the findings

We did not observe any effect of this vaccine regimen on IVF outcomes or pregnancy rates. This underscores worldwide data and physiologic reasoning that there should not be significant effects of this COVID vaccine on developing gametes quality and function and is a good option for those trying to conceive.

Trial registration number

LND 0031-21
O-221 The concept of ‘family' in the era of assisted conception : a sociolinguistic analysis
Authors: Lockwood G
Published: June 2022

Study question

Should current societal concerns about how the application of ART has resulted in evolving family structures influence clinical and legislative practice 

Summary answer

Concerns about the role of ART in ‘threatening' historical norms need to be addressed as they cause distress and fail to support valid life choices

What is known already

Techniques of assisted conception including gamete donation, cryopreservation and surrogacy have enabled many individuals to access unconventional and different family structures. ART modalities that were initially concerned with achieving a child for a hetero-normative couple are now widely accessed by individuals and couples with alternative family structures. Current societal concerns oscillate between anxieties about inappropriate access and recourse to fertility treatments, demographic evidence of declining fecundity and geo-political pressures to optimize social structures.   

Study design, size, duration

A historical review was undertaken of family structures during the historical epoque since records are available of family size, maternal mortality, generational divergence and ‘un-official' family formation. Consideration of literary sources including foundlings, child ‘gifting', adoption and abandonment was involved. Assessment of current cultural norms including Census data information on household composition was accessed. Semi-qualitative interviews with individuals wishing to access fertility treatment with donor gametes.. 

Participants/materials, setting, methods

On-line access to records, census data, historical reviews etc.

Main results and the role of chance

In the post-industrial era a significant proportion of children lived in families to which they had no genetic link. ‘Excess' children were ‘gifted' to relatives, ‘sent' to live with individuals who offered the opportunity of a ‘better' life or who simply required  a servant, or a companion or an heir. Quantitatively this is no different from the current situation in the UK  where half of all young children live in a household in which they are not genetically linked to one of the adults who care for them. Deferred marriage or coupledom and age-related subfertility have increased the use of donor eggs. Single women and same sex female partners can now freely access donor sperm. it could be concluded that the current ‘moral panic'about the status of ‘the family' is misguided and historically inaccurate.Contemporary family structures  are not so different from what has preceded them, even though the ways in which these longed for  children arrive may be nove.l   

Limitations, reasons for caution

These data refer to UK sources and may not be universalisable

Wider implications of the findings

Practitioners of ART and allied professionals who support their work need to be aware of the needs and entitlements of individuals who seek to form families by accessing ART.

Trial registration number

Not applicable
P-741 The COVID-19 pandemic: is there any impact on male fertility?
Authors: Sallem A , Garrouche S , Ben Fredj M , Kooli R , Boughzala I , Boussabbeh M , Ammar O , Mehdi M
Published: June 2022

Study question

Is there any impact of the pandemic period on semen parameters?

Summary answer

Both total and progressive sperm motility as well as sperm morphology were impaired during COVID-19 pandemic.

What is known already

Male fertility could be affected by many environmental conditions. The COVID-19 pandemic has led to many dramatic consequences on human lives (psychological, financial level…). However, little information is available on the impact of the emergent COVID-19 on male fertility.

Study design, size, duration

This was a cohort study comparing semen parameters before and during the two first COVID-19 waves in infertile Tunisian patients.

Participants/materials, setting, methods

Were included in the current study 90 patients followed in the consultation of the department of Cytogenetics and Reproductive Biology (Monastir, Tunisia) for hypofertility. Each of the included patients has already a spermogram before the COVID-19 pandemic and a spermogram during the COVID-19 pandemic allowing the comparison of semen parameters for each patient so that he was considered as his own control. Patients who received medication (antibiotics, antioxidants…) were excluded from the current study.

Main results and the role of chance

Among standard semen parameters, we have shown a significant decrease in both total and progressive sperm motility during COVID-19 pandemic (p < 0.0001 and p = 0.001 respectively). The observed decrease 30 min after ejaculation was maintained 2 hours and 4 hours after ejaculation. Furthermore, we observed an impairment in sperm morphology. Indeed, the percentage of morphologically abnormal spermatozoa raises from 90.99±7.37% to 93.67±4.54% (p < 0.0001). The remaining semen parameters was similar between the two compared timepoints except a slight decrease in sperm count during the pandemic (p = 0.079).

Multivariate analysis didn’t show among clinical and epidemiological characteristics any associated factor with the observed decrease in semen quality.

Limitations, reasons for caution

The included patients didn’t have any COVID-19 symptoms on the day of sperm collection. However, as we have no proof of negative PCR test, the observed impairment in semen quality could be not only the consequence of psychological stress but may be also induced by a latent infection.

Wider implications of the findings

Even in patients with no proof of COVID-19 infection, the pandemic seems to have a real impact on hypofertile men as sperm motility and morphology were significantly impaired. It would be preffered to control semen parameters away from such period before referring patients to assisted reproduction.

Trial registration number

Not Applicable
O-230 Newer non-invasive modalities in fibroid management
Authors: Dhillon Pai R
Published: June 2022

Fibroids are highly prevalent and may cause abnormal menstrual bleeding, pain, mass in pelvis, infertility or pregnancy complications. For years, surgery such as myomectomy or hysterectomy have been the main treatment option. However, the high costs, morbidity and often long hospitalization and recovery time have driven the need for more conservative options.

Today, there is a paradigm shift in our approach to the management of fibroids.

Depending on the age of the woman, number and location of fibroids, symptoms and her desire for fertility, one can choose a suitable option. Medical options such as progestin’s, GnRh analogues, GnRH antagonists, selective progesterone receptor modulators like ulipristal, mifepristone, aromatase inhibitors or the use of an intra uterine progestin releasing device give good relief of symptoms and temporarily shrink the fibroid.

Many high intensity focussed ultrasound options with or without MRI guidance are now available. Uterine artery embolization has stood the test of time.

Innovative new techniques such as transvaginal doppler guided vascular clamp, intrauterine ultrasound guided radio frequency ablation, percutaneous microwave ablation are now available to treat this common, often symptomatic and recurrent problem.
O-289 Effects of Taxol on the developmental potency of human and mouse GV oocytes
Authors: Watanabe S , Atsushi T
Published: June 2022

Study question

Is it possible to improve the quality of oocytes with delayed maturation or aging?

Summary answer

Yes, modification of oocyte microtubules at the germinal vesicle (GV) stage inhibits abnormal chromosome separation in in-vitro maturation and enhances early cleavage.

What is known already

Oocyte aging is characterized by an increase of aneuploidy and a decrease of the developmental potency with maternal ages, which will results in low frequencies of the blastocyte formation and implantation. In addition to the 1st meiotic division, it has become clear that the 2nd meiotic division also significantly contributes to aneuploidy production which is followed by pre-implantation embryo loss. However, no method for overcoming the oocyte aging has been established.

Study design, size, duration

Human GV oocytes with delayed maturation obtained from consented female patients were used. Mouse GV oocytes collected from 9-15 month old ICR mice were also used. After exposure to Taxol for 1hr, the oocytes were matured in vitro and examined cytogenetically and cytologically at the GV stage, the MII stage and pronuclear stage.

Participants/materials, setting, methods

RNA transcripts of the GV stage oocytes were compared before and after Taxol exposure with microarray in both species. Chromosome aberrations at the MII stage and blastocyst formation rate were examined in human IVM oocytes. Mouse IVM oocytes were evaluated on 2nd polar body (PB) extrusion and O2 consumption (CRAS3.0, Crino Co Inc.) at the pronuclear stage after parthenogenetic activation with electro-stimulation.

Main results and the role of chance

After Taxol treatment, premature chromosome separation was significantly reduced from 96% to 7% and the blastocyst formation rate increased from 3% to 16% in human. In mice, normal PB extrusion rate increased from 29% to 92%, and oxygen consumption was higher in some pronuclear oocytes. In human GV oocytes, taxol treatment altered gene expression only in a few factors related to chromosome attachment and segregation. On the other hand, in mouse GV oocytes, gene expression was significantly altered by oocyte aging and Taxol treatment.

Limitations, reasons for caution

The present results were obtained in human GV oocytes with delayed maturation which were collected after PMS and HCG administrations. It is not clear whether Taxol is effective on the GV oocytes collected before HCG administration.

Wider implications of the findings

Our results indicate that the combination of biochemical modification and IVM in the GV stage may be able to restore the aging of human oocytes.

Trial registration number

not applicable
O-303 Recurrent pregnancy loss is associated with unfavorable composition of endometrial and vaginal microbiota
Authors: Peuranpää P , Holster T , Saqib S , Kalliala I , Tiitinen A , Salonen A , Hautamäki H
Published: June 2022

Study question

Does endometrial or vaginal microbiota composition associate with recurrent pregnancy loss (RPL)?

Summary answer

Reduced relative abundance of Lactobacillus crispatus in endometrium and increased relative abundance of Gardnerella vaginalis in endometrium and vagina were associated with RPL.

What is known already

Dysbiotic vaginal microbiota associates with sporadic miscarriage and the depletion of lactobacilli in the endometrium has been associated with poor IVF outcomes, but their association with RPL has not been studied.

Study design, size, duration

A nested case-control study of 47 women investigated for RPL and 39 healthy control women without a history of pregnancy loss were recruited between March 2018 and December 2020 at a university hospital. Endometrial biopsies and vaginal samples were collected from the women 6-8 days after positive ovulation test result.

Participants/materials, setting, methods

The endometrial and vaginal microbiota compositions, analysed using 16S rRNA gene amplicon sequencing, were compared between RPL women and controls, and between individual vaginal and endometrial samples. The mycobiota composition was analysed using internal transcribed spacer 1 amplicon sequencing for a descriptive summary. The models were adjusted for BMI, age, and parity. False discovery rate -corrected P-values (q-values) were used to define nominal statistical significance at q< 0.05.

Main results and the role of chance

Lactobacillus crispatus was less abundant in the RPL women’s endometrial samples compared to controls (mean relative abundance 17.2% vs. 45.6%, q= 0.04). Gardnerella vaginalis was more abundant in RPL women than in controls in both endometrial (12.4% vs. 5.8%, q< 0.001) and vaginal samples (8.7% vs. 5.7%, q< 0.01). The individual vaginal and endometrial microbial compositions correlated strongly (R = 0.85, P< 0.001). Fungi, mostly Candida albicans, were detected in 22% of the endometrial and 36% of the vaginal samples.

Limitations, reasons for caution

Limitations include the transcervical sampling technique with a potential risk of cervicovaginal contamination of endometrial samples, the loss of a proportion of the samples from statistical analyses due to their low read count, and the presentation of the 16S rRNA sequencing results in relative rather than absolute abundances

Wider implications of the findings

Our findings suggest that an unfavorable endometrial microbiota may be a novel risk factor for RPL. Further studies should elucidate the mechanisms, how microbiota may predispose to RPL, possibilities to modify microbiota by treatments, and the effects of such treatments on the success of future pregnancies in couples having RPL.

Trial registration number

not applicable
O-304 An altered microbiota in the lower and upper female reproductive tract of women with recurrent spontaneous abortion
Authors: Liu F , Yang S , Yu Y , Li R
Published: June 2022

Study question

Whether the vagina, cervical and uterine microbiota have changed in women with recurrent spontaneous abortion (RSA).

Summary answer

An altered microbial biodiversity in the vagina, cervix and uterine lavage fluid in the RSA group was observed.

What is known already

An obstacle to pregnancy, RSA can be caused by a variety of factors, including genetics, uterine anomalies, autoimmune diseases, and infection. However, the current understanding of the etiology of RSA is still lacking, and half of cases have unknown causes. Studies have demonstrated that two genera (Atopobium and Prevotella) exhibited significantly greater abundance. The characteristics of the microbiota harbored in vaginal secretions in RSA have been revealed but less attention has been given to the cervical and uterine microbiota in RSA patients.

Study design, size, duration

Samples were collected (RSA group: n = 25; Control group: n = 25) in Peking University Third Hospital from December 2020 to June 2021.

Participants/materials, setting, methods

Women with two or more consecutive spontaneous abortions were included. Microbiota compositions were acquired and analyzed by16S rRNA sequencing and Th1/Th2/Th17 inflammation cytokines level of uterine flush fluid were tested.

Main results and the role of chance

Beta diversity was significantly higher in the RSA group than in the control group in the vaginal microbiota (p = 0.036), cervical microbiota (p = 0.010) and microbiota from uterine lavage fluid (p = 0.001). In addition, dramatic decreases in IFN-γ and IL-6 cytokine levels were observed in the RSA group. In conclusion, our data suggested altered microbial biodiversity in the vagina, cervix and uterine lavage fluid in the RSA group. Moreover, the microbiota composition differed markedly from the lower genital tract to the uterine cavity, and the microbiota in the uterine cavity also distinctly varied between endometrial tissue and uterine lavage fluid in the RSA group (p = 0.001).

Limitations, reasons for caution

Due to the limitations of 16S rRNA sequencing, only relative rather than absolute abundances of the microbiota were obtained.

Wider implications of the findings

Our study suggested that significant alterations in the microbial profile of the vagina, cervix and uterine cavity were present in RSA patients, and the key microbiota in different locations of female reproductive tract involved in pathogenic processes were different, which indicated that different treatments should be considered in clinics.

Trial registration number

Not applicable
P-009 A novel sperm selection technique using a progesterone gradient
Authors: Romualdo P , Fasano C , D'Andolfi G , Tozza G , Gualtieri R , Dale B
Published: June 2022

Study question

Is it possible to select spermatozoa using progesterone gradients? Does chemotaxis improve sperm selection compared to a classical density gradient or swim up?

Summary answer

Spermatozoa selected using progesterone gradients yield higher percentages of high quality spermatozoa with lower DNA fragmentation and better morphology compared to classic sperm selection.

What is known already

There are few studies which aim at improving sperm selection, even though it is known to be a vital parameter for IVF success. DNA integrity is fundamental for embryo development although it is currently underestimated.

Progesterone has been suggested to be a major chemoattractant for spermatozoa in the fallopian tubes and therefore using gradients in a clinical setting may lead to a more physiologically improved selection method.

Study design, size, duration

19 normo spermic patients (according to W.H.O. parameters) were selected. After standard semen analysis the sample was split in two halves. One half was treated using classical swim up and the remaining half was treated on a progesterone gradient. The treated samples were then tested for fragmentation, motility and morphology. Sperm motility was analyzed using SCA computer system. DNA fragmentation using the TUNEL assay.

Participants/materials, setting, methods

The patients enrolled in the study were all normospermic. The results were compared using ANOVA one way. Progesterone gradients was obtaining using an horizontal density gradients with added progesterone.

Main results and the role of chance

The percentages of rapid progressive spermatozoa were 12,89 ± 4,50%; 56,37 ± 13,61% and 29,84±12,52% in the untreated sample (US), treated sample (TS) and control group (C)respectively; p < 0,005. Normal sperm morphology rates were 3,474 ± 1,46; 5,500 ±1,57 and 3,684 ± 1,15 in US, TS and C respectively. Sperm fragmentation rates were 9,77 ± 9,012,50 ± 2,66 and 7,94 ± 6,68 in US, TS and C respectively.

Limitations, reasons for caution

The number of normospermic patients enrolled in the study was limited. Moreover, the sperm concentration obtained in this kind of selection is lower than in the conventional techniques and therefore may be harder to use in patients with a very low sperm count.

Wider implications of the findings

Our data shows a significant improvement in sperm selection using chemotaxis gradient compared to the control group by all the parameters considered in this study. Chemotaxis gradients may be useful in a clinical setting to improve spermatozoa selection, lowering the rates of sperm with damaged DNA potentially increasing embryo quality.

Trial registration number

Not applicable
P-025 Freezing of the semen sample by the patient at home is clinically effective
Authors: Pujal B , Montalvo V , Massó J , Garcia-Faura Á , Garcia F , Marqués B , López-Teijón M
Published: June 2022

Study question

To evaluate the efficacy of the method of freezing the seminal sample at home by the patient himself.

Summary answer

The freezing of the seminal sample at home is an effective method that brings the clinic closer to the patient, eliminating travel and stress.

What is known already

Seminal sample freezing for reproductive purpose is indicated in different situations: preservation of fertility, before a vasectomy, in patients with dysfunctions and problems obtaining a sample, and for those patients who cannot be present at the time of the oocyte retrieval.

In many of these cases, the patient is under pressure to obtain the sample, in a clinical environment and at an exact time. Many experience this as a stressful situation, which in some cases forces them to travel if they live far from the clinic.

Study design, size, duration

In 2017, the home freezing protocol was validated with volunteers who obtained a semen sample at the clinic. After verifying that the quality of the sample was the same as if it were frozen by an expert biologist, we began offering this technique to our patients in 2019. We compared the results of the DO cycles with a control group to determine the efficacy of the protocol.

Participants/materials, setting, methods

Until December 2021, a total of 18 patients have used the Freezing Kit at home, 14 were residents in other EU countries, and 4 residents in Spain, but far from the clinic. As a control group, we used 608 oocyte donation cycles with a frozen sample.

Patients who decided to freeze their sample at home receive a kit consisting of a travel tank with nitrogen vapors, a box with all the necessary material, and instructions.

Main results and the role of chance

Regarding the results of the 18 patients who have used the kit to freeze samples for their oocyte donation cycles, they have a mean of 44±7.1 years, and the samples had a post-thaw concentration of 48±30.5 M /ml with a mean mobility of 21.18±14.6%.

No statistically significant differences were found in the fertilization rate compared to the control group (76.4±15.1% vs 77.8±18.3%; p = 0.485), blastocyst rate (61%±10 .5% vs 56.1±16.4%; p = 0.387) nor clinical pregnancy rate (62.8% vs 47.6%; p = 0.251).

The fact of having a live birth from a sample frozen by the patient himself has just demonstrated the safety and usefulness of this novel freezing protocol.

Limitations, reasons for caution

The number of patients who have undergone this process is limited due to the nature of the treatment.

Wider implications of the findings

These results demonstrate the clinical utility of this freezing method for certain patients, where travel to the clinic may be inconvenient. Demonstrating that it is an effective method of cryopreservation of the seminal sample that brings the clinic closer to the patient, eliminating trips and stress.

Trial registration number

not applicable
P-026 Lack of association between single polymorphic variants of the mitochondrial nicotinamide adenine dinucleotide dehydrogenase 3, and 4L (MT-ND3 and MT-ND4L) and male infertility
Authors: Dahadhah F , Saleh Jaweesh M , Salim Al Zoubi M , Issam Abu Alarjah M , Eid Hammadeh M , Amor H
Published: June 2022

Study question

Is there any association between male infertility and the polymorphic variants of Mitochondrial Nicotinamide Adenine Dinucleotide Hydride dehydrogenase (NADH) Subunit 3 and 4L (MT-ND3 and MT-ND4L)?

Summary answer

The genotypes and allele frequencies of the study population have shown a lack of statistically significant association between MT-ND3 and MT-ND4L SNPs and male infertility.

What is known already

Male infertility is a multifactorial condition associated with different genetic abnormalities in at least 15%–30% of cases. The rate of mutations in the mtDNA is high due to the lack of histones and DNA repair mechanisms. Moreover, mtDNA is more vulnerable compared to nuclear DNA due to the lack of an efficient repairing system and high exposure to oxidative species produced by the mitochondria. Eventually, this leads to abnormal sperm function, structure and even infertility.

Study design, size, duration

Prospective study was carried out between 2018 and 2019.

112 semen samples were collected in this study.

Participants/materials, setting, methods

The present study was carried out at the department of Obstetrics and Gynecology, University of Saarland,Germany. After semen analysis, samples were purified. Mitochondrial DNA was extracted using a QIAamp DNA Mini Kit, then, the mitochondrial DNA was amplified by using the REPLI-g Mitochondrial DNA Kit. Polymerase chain reaction (PCR) was used to amplify MT-ND3 and MT-ND4L genes. After that samples were purified and sequenced using the Sanger method in the Microsynth Seq Lab in Germany.

Main results and the role of chance

Eight single nucleotide polymorphisms (SNPs) in MT-ND3 (rs2853826, rs28435660, rs193302927, rs28358278, rs41467651, rs3899188, rs28358277 and rs28673954) and seven SNPs in MT-ND4L (rs28358280, rs28358281, rs28358279, rs2853487, rs2853488, rs193302933 and rs28532881) were detected and genotyped. The genotypes and allele frequencies of the study population have shown a lack of statistically significant association between MT-ND3 and MT-ND4L SNPs and male infertility. Besides, no statistically significant difference was found between the asthenozoospermia, oligozoospermia, teratozoospermia, asthenoteratozoospermia, oligoasthenoteratozoospermia and oligoteratozoospermia subgroups of subfertile males. However, rs28358278 genotype of the MT-ND3 gene was reported in the subfertile group but not in the fertile group, which implies a possible role of this SNP in male infertility.

Limitations, reasons for caution

The size number of the study samples.

Wider implications of the findings

Further studies are required to evaluate these findings. Moreover, the subfertile individuals who exhibit a polymorphism at rs28358278 require further monitoring and evaluation.

Trial registration number

Basic Science
P-060 Bubble-shaped Acrosome: Novel phenotype of Teratozoospermia Caused by Mutation in Actin-like 7A
Authors: Dai J , Chen Y , Li Q , Zhang T , Gong F , Lu G , Lin G
Published: June 2022

Study question

What’s the etiology of severe teratozoospermia characteristic as bubble-shaped acrosome (BSA)?

Summary answer

Severe teratozoospermia characterized as BSA caused by mutation (c.1024G>A) in actin-like 7A (ACTL7A).

What is known already

Teratozoospermia is a common cause of male infertility, defined by having a proportion of morphologically normal sperm at less than 4%. It exhibits aberrant sperm phenotypes in the head, neck, midpiece, and endpiece of sperm. Teratozoospermia with ephalic abnormalities are among the most severe and characteristic sperm defects. Some genetic factors are reported to be associated with ephalic abnormalities such as globozoospermia and macrozoospermia. However, other phenotypes and the causative genes of ephalic abnormalities, especially in acrosomal structure, and were largely unknown.

Study design, size, duration

Severe teratozoospermia were recruited from the Reproductive and Genetic Hospital of CITIC-Xiangya from Jan 2019 to Dec 2021.

Participants/materials, setting, methods

Whole-exome sequencing analysis was used to analyze the genetic factor of man. An Actl7a-mutated mouse model was generated by CRISPER-Cas9. Transmission electron microscopy was used to detect the abnormality of ultrastructure during acrosome biogenesis. Immunostaining was used to analyze the localization of ACTL7A and PLCζ. Immunoprecipitation followed by liquid chromatography-mass spectrometry (LC-MS) was used to select the differentially expressed proteins. ICSI with calcium ionophore exposure was performed in couple with ACTL7A mutation.

Main results and the role of chance

We found a man with severe teratozoospermia characterized as BSA carrying a mutation (c.1024G>A) in ACTL7A. Homozygous Actl7a-mutated male mice were sterile, and all of sperm showed acrosomal abnormalities. During acrosomal biogenesis, it detected the acrosome detach from the nuclear in Actl7a-mutated mice. Furthermore, mutant ACTL7A failed to attach to the acroplaxome and was discharged by cytoplasmic droplets, which led to the absence of ACTL7A in mature sperm. The mutant sperm failed to activate the oocyte, and PLCζ discharge accompanied by ACTL7A was observed, leading to total fertilization failure (TFF). Immunoprecipitation followed by LC-MS showed that several differentially expressed proteins participate in acrosome assembly and actin filament organization. Furthermore, assisted oocyte activation by calcium ionophore exposure successfully overcame TFF in a couple with an ACTL7A mutation.

Limitations, reasons for caution

More cases are needed to demonstrate the relationship between mutation and phenotype.

Wider implications of the findings

Our study defined a novel phenotype of the acrosomal abnormality characterized as BSA and revealed the underlying mechanism of mutation in ACTL7A and provided a genetic marker and a therapeutic option for male infertility.

Trial registration number

Not Applicable
P-161 The role that endogenous oleic acid synthesis plays in cell proliferation and expression of folliculogenesis markers in ovarian granulosa cells
Authors: Fayezi S , Lange C , Jauckus J , Rehnitz J , Nguyen X.P , Strowitzki T , Germeyer A
Published: June 2022

Study question

Does a deficiency of endogenous oleic acid synthesis affect cell proliferation and the expression of folliculogenesis markers in the ovarian granulosa cell line COV434?

Summary answer

Endogenous oleic acid synthesis supports cell proliferation and the expression of folliculogenesis markers, which are possibly linked to oocyte maturation.

What is known already

The endogenous synthesis of the monounsaturated fatty acid oleic acid from saturated fatty acids is catalysed by stearoyl-coenzyme A desaturase (SCD). Previous studies have suggested that lipid monounsaturation may promote steroidogenesis and oocyte development. However, the potential role of endogenous oleic acid synthesis in folliculogenesis remains virtually unexplored. Autophagy related 14 (ATG14), growth differentiation factor 9 (GDF9), signal transducer and activator of transcription 3 (STAT3) and forkhead box protein O1 (FOXO1) play essential roles in follicular development via promotion of proliferation and maturation of ovarian granulosa cells.

Study design, size, duration

Immortalized granulosa cells (COV434) were cultured, treated with a specific SCD inhibitor (SCDi) and supplemented with or without oleic acid for 48 h. The cells were cultured for a further 24 h in normal media. The cell culture medium was then discarded and the cells were washed before a viability assay or RNA extraction was performed. RNA extraction was followed by cDNA synthesis and real-time PCR assessment. All experiments were conducted three times in duplicate.

Participants/materials, setting, methods

Cell proliferation was determined using a crystal violet viability assay. The expression of the folliculogenesis markers ATG14, GDF9, STAT3 and FOXO1 was analysed using TaqMan real-time PCR.

Main results and the role of chance

The cell proliferation rate was considerably lower in SCDi-treated cells compared to untreated control cells (-50%, p < 0.01), and the effect was fully recovered by adding oleic acid to the SCDi-conditioned cells. SCDi treatment led to significant decreases in ATG14 (-90%), GDF9 (-86%), STAT3 (-62%) and FOXO1 (-93%) (p < 0.01) compared with the baseline control levels. Oleic acid was found to partially, but significantly, suppress the inhibitory effect of SCDi on the expression of the folliculogenesis markers. The rescue effect of oleic acid was significantly stronger after an extended culture time following SCDi treatment.

Limitations, reasons for caution

Further studies need to be performed, possibly with primary cell cultures, to confirm these effects.

Wider implications of the findings

The involvement of the endogenous synthesis of oleic acid in ovarian granulosa cell functionality is relevant to reproductive disorders that affect oogenesis.

Trial registration number

not applicable
P-291 Positive effects of inactivated blood serum in stabilizing the activity of antioxidants in embryo-free culture media
Authors: Ramirez-Dominguez L.B , Jimenez Medina I , Maldonado-Rosas I , Villar-Muñoz L.G , Saleh R , Agarwal A
Published: June 2022

Study question

Does addition of blood serum to embryo culture media supplemented with antioxidants (AOXs) helps maintain the AOX activity under in vitro culture conditions?

Summary answer

The addition of inactivated blood serum (IBS) before AOX supplementation resulted in AOX activity stabilization in embryo-free culture media (EFCM) for up to 48 hours.

What is known already

Supra-physiological levels of reactive oxygen species (ROS) in embryo culture medium (ECM) can disturb embryo development possible due to oxidative stress. Hence, AOX supplementation of ECM has been used to counteract the potential negative effects of high ROS on gametes and embryos. However, rapid consumption of AOXs via interaction with ROS may reduce the AOX protection capacity particularly during extended culture conditions. The aim of this study is to investigate the efficacy of IBS in stabilizing the AOX activity of a combination of AOXs (EmbryORP®) when added to EFCM.

Study design, size, duration

In this prospective study, oxidation reduction potential (ORP) was assessed in EFCM by MiOXSYS System (UAB Caerus Biotechnologies, Vilnius, Lithuania). The ORP results reflect the overall balance between oxidants and antioxidants. ORP levels in follicular fluid obtained from dominant follicles of 40 oocyte donors were used as control.

Participants/materials, setting, methods

All dishes containing culture medium (GlobalTotal®, LifeGlobal®, Connecticut, US) were incubated for 48 hours in 8% CO2, 20% O2, 37 °C. Culture dishes were divided into four groups: Group A: EFCM without AOX, Group B: EFCM+AOX, Group C: EFCM+AOX+IBS, and Group D: EFCM+IBS. Levels of ORP were assessed in EFCM before AOX supplementation (0), immediately after adding AOX (1) and 48 hours following AOX supplementation (2). ORP results are expressed as millivolts (mV).

Main results and the role of chance

At 48 hours of culture, ORP value in group C (96.7 mV) was significantly lower than group A (345 mV; p = 0.00001), and was found to be similar to the physiological ORP values found in follicular fluid from donors (89 mV±23.6 mV; p > 0.05). Also, ORP value in group C (96.7 mV) was lower than group B (210 mV; p = 0.67) and group D (280.1 mV; p = 0.11).

Limitations, reasons for caution

This study only tested the efficacy of IBS as AOX stabilizer in EFCM over 48 hours. The efficacy and potential risks associated with the use of IBS during actual embryo culture were not examined.

Wider implications of the findings

The use of AOX stabilizers may represent an important novel finding to maintain sufficient AOX protective capacity during extended culture conditions.

Trial registration number

10112021-AOX
P-293 Euploidy results through niPGT techniques
Authors: Fernández Díaz M , De Luxán Delgado B , Atienza de Nava S , Alonso Cuevas B , López Cañal P , Muñoz Oreña P , García Álvarez E , Gayo Lana A
Published: June 2022

Study question

Which are the euploid results using niPGT?

Summary answer

Our niPGT (non invasive PGT) results are similar to the one obtained through traditional PGT-A

What is known already

Recent studies have shown the presence of embryo cell-free DNA (cfDNA ) in spent blastocyst medium (SBM), opening a new era of possibilities for non-invasive preimplantation genetic testing for aneuploidy (niPGT-A).

Study design, size, duration

We performed a retrospective study including 80 patients of IVF between August 2020 and December 2021, among them, 60 used their own egg, and 20 donor eggs. 237 embryos were analysed through niPGT.

Participants/materials, setting, methods

Embryos were cultured in a Geri incubator (Merck) up to day 4, and then individually cultured in 10 µl drops of CCSS (Fujifilm) until day 6 in a bench-top Minc (Cook). At day 6, after at least 40 hours in culture, blastocysts were vitrified and medium collected in sterile PCR tubes. After collection, medium was immediately frozen and sent for sequencing to our reference laboratory (Igenomix, Spain).

Main results and the role of chance

Euploidy ratio by niPGT analysis was significantly lower in patients older than 40 years (15%) comparing with both 35-40 year patient (48%) and younger than 35 years old patients (50%) (p < 0.0001). The euploidy ratio showed by embryo-patient oocytes (37%) was also significantly lower than donor oocytes (55%) (p < 0.5). Our results showed that the group 7 chromosomes (19 and 20) were significantly more altered in patients older than 35 years old (p < 0.5).Non informative results were only obtained in own egg embryos (7%), while all the embryos from donor eggs showed informative results (either altered or euploid).

Limitations, reasons for caution

This is a retrospective study. niPGT is a new technique for euploidy screening, therefore more studies involving evolutive pregnancy ratio are necessary for its optimal clinical application replacing biopsy PGT.

Wider implications of the findings

Our results strongly support the use of cfDNA as niPGT technique as a potential marker of embryonic genetic status, that could even replace traditional techniques as invasive PGT in the future.

Trial registration number

No aplica
P-310 The last journey: 448KHz electrostimulation and its effects on the endometrium of women going through menopause
Authors: Irollo A.M , Bracone G , Renzi A , Gangale M.F , Tartaglia M.L , Calabrese G , Irollo A.M , Criscuolo C , Tavoletta B , Pellegrini M , Ghelardi C , De Simone M , Rosa M. Di , Di Nocera A , Stortini E
Published: June 2022

Study question

Could 448KHz electrostimulation be beneficial in the endometrial preparation for transfer in pre-menopausal or menopausal women?

Summary answer

Few women had adequate endometrium (8.3%) with standard treatments, which increased to 54.1% when combined with aspirin-sildenafyl, while when combined with 448KHz is over 75%

What is known already

We know that electrical stimulation is used in tissue engineering as a new regenerative medicine approach, which promotes cell proliferation and differentiation. In addition, it improves cell membrane permeability and cell metabolism.

Study design, size, duration

In the retrospective clinical study a total of 96 patients were considered, all in premenopausal or menopausal state, with inadequate endometrial response to common therapies and failure to transfer. In the following cycle, attempts were made to optimize the level of endometrial preparation by various approaches.

Participants/materials, setting, methods

The instrument used for electrostimulation was INDIBA. The 96 patients were divided into 4 groups: group A received estrogen therapy and 448KHz electrostimulation, group B received estrogen therapy, group C received estrogen therapy, aspirin and sildenafyl support, group D received estrogen therapy, aspirin-sildenafyl support and 448KHz electrostimulation.

Main results and the role of chance

Electrostimulation was found to be beneficial in group A (with electro-stimulation) where 75% of patients had an endometrium suitable for transfer compared to group B (without electro-stimulation) with only 8.3%. Similarly, group C (without electro-stimulation) had 54.1% of patients with a suitable endometrium, while group D (with electro-stimulation) had 87.5%.

Limitations, reasons for caution

The limitations are related to the nature of the study, which is retrospective, so although the starting conditions were evaluated to reduce bias, we suggest confirmation with a prospective randomized clinical trial to validate these data, which will be our next objective.

Wider implications of the findings

The common therapeutic approach of endometrial preparation, when combined with electrostimulation, leads to a drastic decrease in cancelled cycles due to an inadequate endometrium.

Trial registration number

not applicable
P-316 Effect of nanoparticle-mediated delivery of miRNA 503 on the apoptosis of ovarian endometriosis cells
Authors: Slahi N , Mehdizadeh Kashi A , Tehermanesh K , Pirhajati Mahabadi V , Chaichian S
Published: June 2022

Study question

Is effective miRNA 503 conjugation with PLGA nanoparticle in apoptosis of human endometriosis cells?

Summary answer

Cell death increased with increasing the concentration of miRNA; thus, it can be suggested as a treatment for endometriosis.

What is known already

MicroRNAs (miRNAs) regulate post-transcriptional gene expression and play a crucial role in proliferation, differentiation, and apoptosis, which are key to the diagnosis of endometriosis.miRNA 503 causes apoptosis and is genetically inhibited in endometriosis patients. Therefore, in this study, we decided to introduce this miRNA into the endometriosis cell using PLGA nanoparticles in the laboratory

Study design, size, duration

The cells were divided into the five groups of control and four doses (25, 50, 75, and 100 µm) of miRNA 503/PLGA. The cells were then treated with the optimum dose obtained, and survival rates were compared with and without nanoparticles. The experiments were performed at 12, 24, 48, and 72 hours. Viability and apoptosis were evaluated by the MTT assay and Annexin Kits. Data were analyzed by one-way analysis of variance.

Participants/materials, setting, methods

In this study, endometriosis cells were isolated from women with severe endometriosis (DIE) and digested by the enzymatic method (40 µg/ml DNAase I and 300 µg/ml collagenase type 3). PLGA-based nanoparticles were synthesized and characterized. The size of sole PLGA nanoparticles was about 60 nm, and that of PLGA/miRNA was about 70 nm. Poly lactic-co-glycolic-based nanoparticles were used as vector carriers for miRNA 503 transfection in endometriosis cells.

Main results and the role of chance

Based on the MTT assay results, the optimal dose of miRNA 503 / PLGA was 75 µm, at which the viability of ECSCs was 52.6%±1.2 (P ≤0.05), and the optimal time was 48 h. The apoptotic rates of ECSCs treated with PLGA/miRNA503 (34.75±4.9%) were significantly higher than those of ECSCs treated with PLGA alone (3.35±2.58%) (P ≤0.05).

Limitations, reasons for caution

Cell mortality in vitro and animal survival after cell injection were among the limitations of the study.

Wider implications of the findings

miRNAs are involved in cell survival, proliferation, angiogenesis, and apoptosis, and therefore, they are effective in the pathogenesis of endometriosis . Viral and non-viral vectors are used for miRNAs transmission. Non-viral gene delivery vectors, such as inorganic nanoparticles and liposomes, have been used in recent years

Trial registration number

not applicable
P-351 Medical treatment for missed abortion- difference in gestational age based on menstrual dating and on ultrasound criteria, and the correlation to failure
Authors: Gluck O , Barber E , Sagiv R
Published: June 2022

Study question

Does the difference in gestational age based on menstrual dating and on ultrasound criteria correlate with failure rate of medical treatment in missed abortions?

Summary answer

In missed abortions, the difference in gestational age based on menstrual dating and on ultrasound is inversely correlated with failure rate of medical treatment.

What is known already

A number of factors have been reported as predictors for Misoprostol treatment failure, including assisted reproductive treatment, primigravity, and vaginal bleeding prior to treatment.

Increased rates of failure with medical management were also reported with delaying treatment after diagnosis.

Study design, size, duration

This was a retrospective cohort study.

Overall, 778 patients who underwent medical treatment with Misoprostol for missed abortion at our institute, between 07/2015 and 12/2020 were included.

Participants/materials, setting, methods

Inclusion criteria were either anembryonic gestation or embryonic death, as long as pregnancy age, as measured by transvaginal sonography , correlated with 12 weeks gestation or less.

Exclusion criteria included incomplete or septic abortions, multiple pregnancies, and cases that had either irregular period, missing data, or were lost to follow up.

The primary outcome was treatment failure, defined as any need for surgical intervention (elective or emergency Dilation & curettage, or operative hysteroscopy).

Main results and the role of chance

Five hundred and eighty two patients (74.9%) had successful medical treatment, while 196 (25.1%) required surgical intervention.

The gestational age based on menstrual dating and on ultrasound difference (in weeks) was 2.6±1.4 in the success group and 3.1±1.6 in the failure group (p < 0.001).

On multivariant regression analysis, gestational age based on menstrual dating and on ultrasound difference was found to be independently correlated with treatment failure (aOR=1.24, CI 95% (1.01-1.51), p = 0.03).

Limitations, reasons for caution

We did not analyze long term consequences, such as the amount of bleeding, and time to future fertility.

Moreover, patients who were treated at our institute may have been diagnosed and treated for treatment failure in neighboring hospitals and hence lost to follow-up.

Wider implications of the findings

The current study has shown that the difference between gestational age based on menstrual dating and on ultrasound is directly corelated with medical treatment failure.

This data should be taken into account when counseling patients regarding the optimal treatment for first trimester missed abortion.

Trial registration number

not applicable