bluuebunny/crossref_metadata_2025_split huggingface dataset:
O-289 Effects of Taxol on the developmental potency of human and mouse GV oocytes
Authors: Watanabe S , Atsushi T 
Published: June 2022

Abstract

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

Abstract

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&lt; 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&lt; 0.001) and vaginal samples (8.7% vs. 5.7%, q&lt; 0.01). The individual vaginal and endometrial microbial compositions correlated strongly (R = 0.85, P&lt; 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

Abstract

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

Abstract

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 &lt; 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

Abstract

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

Abstract

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

Abstract

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&gt;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&gt;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

Abstract

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 &lt; 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 &lt; 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