bluuebunny/crossref_metadata_2025_split huggingface dataset:
P–548 High-risk genetic matching on gamete donors: complete genes analysis or genotyping test?
Authors: Molin. Romero M , Yoldi A , Gañán M , Navas P , De. Pico J L , Vaquero Á , Fuente P D l , Ramírez J P , Castilla J A 
Published: August 2021

Abstract

Study question
What carrier screening test is better to reduce the risk of offspring being affected by recessive diseases when genetic matching is performed with gamete donors: complete or targeted genes analysis?

Summary answer
The use of complete genes analysis in the carrier screening of gamete donors reduces the risk of offspring being affected by recessive diseases.

What is known already
Legislative measures and scientific societies alike call for more research to be conducted into recessive diseases in gamete donors, in order to reduce reproductive risk. However, it is still unclear which genes should be studied and what type of data analysis, targeted or nontargeted, should be performed.

Study design, size, duration
This descriptive observational study of 923 oocyte donors and 895 semen donors was conducted from January 2017 to August 2020, at a private gamete bank.

Participants/materials, setting, methods
1818 gamete donors screened by NGS and nontargeted analysis of the variants, the pathogenic variants detected were analysed to estimate the probability of high-risk genetic matching and to determine the results that would have been obtained if the three most commonly used genotyping tests for carriers of recessive diseases in ART had been applied.

Main results and the role of chance
The probability of high-risk genetic matching with gamete donation, screened by NGS and complete genes analysis, was 5.48%, versus the 0.57–2.8% that would have been obtained if the genotyping test had been applied. Of the 1739 total variants found, only 28.69% would have been detected by all three targeted tests considered and 45.66% of the variants would not have been detected by any of them.

Limitations, reasons for caution
The study was not based in the general population, was limited to a population of Mediterranean ethnic origin. In addition, our study only analysed 302 recessive diseases of the 1,300 plus that have been described.
Wider implications of the findings: Our study highlights the considerable heterogeneity of the genotyping tests commonly used in ART, which present significant differences in their ability to detect pathogenic variants. Therefore, the use of genotyping tests for genetic matching is associated with a higher reproductive risk, compared to the use of complete genes analysis.

Trial registration number
Not applicable
P–553 Women with molar pregnancies have a genetic susceptibility to aneuploid miscarriages
Authors: Slim R , Khawajkie Y , Hoffner L , Tan L , Ab. Rafea B , Aguinagua M , Horowitz N S , Ao A , Tan S L , Brown R , Buckett W , Surti U , Hovanes K , Sahoo T , Sauthier P 
Published: August 2021

Abstract

Study question
What causes non-molar miscarriages in women with one hydatidiform mole (HM)?

Summary answer
We found a higher rate of aneuploidies in the non-molar miscarriages of women with HM than in those from women with sporadic or recurrent miscarriages.

What is known already
Women with hydatidiform moles have higher rates of miscarriages and women with recurrent miscarriages have higher rates of moles than women from the general population.

Study design, size, duration
We retrieved archived formalin-fixed paraffin embedded tissues from non-molar miscarriages of patients with one HM and analyzed them for the presence of aneuploidies using single nucleotide polymorphism (SNP)-microarray. We next determined the meiotic origin of the aneuploidies by genotyping the aneuploid non-molar miscarriages along with the parental genomes using microsatellite markers.

Participants/materials, setting, methods
All participants and some of their partners provided written consent to participate in our study, agreed to a blood draw for genotyping analysis, and agreed for us to retrieve their molar and non-molar tissues from various histopathology laboratories for research purposes.

Main results and the role of chance
We demonstrate for the first time that patients with an HM and miscarriages are at higher risk for aneuploid miscarriages [83.3%, 95% confidence interval (CI): 0.653–0.944] than women with sporadic (51.5%, 95% CI: 50.3–52.7%, p value = 0.0003828) or recurrent miscarriages (43.8%, 95% CI: 40.7–47.0%, p value = 0.00002). Genotyping the aneuploid miscarriages and the parental genomes demonstrated that most of the aneuploidies originated from errors in maternal meiosis I or II.

Limitations, reasons for caution
We were able to retrieve only 30 non-molar miscarriages from women with one HM for analysis. Expanding such analysis to a larger and independent cohort of miscarriages from such patients will be important to validate our observations.
Wider implications of the findings: Our data suggest common genetic female germline defects predisposing to HM and aneuploid non-molar miscarriages in some patients.

Trial registration number
Not applicable
P–555 Recurrent pregnancy loss is associated with changes in the pre-pregnant endometrial gland transcriptome
Authors: Pearson-Farr J , Lewis R , Cleal J , Cheong Y 
Published: August 2021

Abstract

Study question
Do endometrial gland factors influence recurrent pregnancy loss?

Summary answer
The endometrial gland transcriptome during the window of implantation is altered in women with recurrent pregnancy loss compared to controls.

What is known already
Secretions from endometrial glands contribute to the uterine environment that supports the attachment and implantation of the embryo in early pregnancy. Studies have attempted to identify an endometrial gene expression pattern associated with recurrent pregnancy loss however, the cellular heterogeneity within the endometrium may obscure important differences in specific cell populations.

Study design, size, duration
An observational study comparing controls and women with recurrent pregnancy loss.

Participants/materials, setting, methods
Endometrial samples were collected during the implantation period of the menstrual cycle from five matched participant egg donor controls and women with recurrent pregnancy loss. Endometrial glands were isolated from fresh endometrial biopsies and RNA sequencing was performed. A differential gene expression analysis and a gene ontology enrichment analysis was performed between egg donor controls and women with recurrent pregnancy loss.

Main results and the role of chance
This study reports a glandular epithelium specific gene expression profile and demonstrates differential gene expression of endometrial glands from women with recurrent pregnancy loss compared to controls. 18 genes were upregulated and 1 gene was downregulated in the endometrial glands from women with recurrent pregnancy loss compared to controls (5% false discovery rate). Biological processes which contain genes that were differentially expressed in women with recurrent pregnancy loss compared to controls include epithelial cell migration and regulation of secretion by the cell.

Limitations, reasons for caution
This is an observational study with a relatively small sample size.
Wider implications of the findings: This study identified differences in gene expression in women with recurrent pregnancy loss that are specifically associated with endometrial glands rather than endometrium as a whole. These differences could be used to identify a perturbed endometrium, isolate causes of recurrent pregnancy loss and develop targeted therapies.

Trial registration number
Not applicable
P–557 Trophectoderm biopsy technique and rate of mosaicism in human blastocysts
Authors: Chan C W , Tee Z Q , Lim A Y X , Lim M W , Lee C S S 
Published: August 2021

Abstract

Study question
Do different trophectoderm biopsy techniques affect mosaicism rate in human blastocysts?

Summary answer
No statistical significance was found between biopsy techniques and mosaicism rate. However, an increase in mosaicism rate was observed when the flicking technique was used.

What is known already
Mosaicism is defined as two or more distinct cell lines within an embryo. Recent advances in Next Generation Sequencing (NGS) technology with an increased sensitivity allows a higher accuracy in quantification of mosaic levels in biopsied cells. The incidence of mosaicism is widely debated as there are many attributing technical and biological factors. Since, trophectoderm biopsy is a technically challenging process, it is crucial to ensure that the both biopsied cells and blastocyst suffers minimal damage during biopsy.

Study design, size, duration
This is a prospective study involving 222 patients (age range= 18–44, mean age= 31.5) who underwent IVF cycles in Alpha IVF, Malaysia from March 2019 to August 2019. Six hundred and sixty-eight (668) of the blastocysts were biopsied on Day 5 (Group 1) while 177 blastocysts were biopsied on Day 6 (Group 2). The blastocysts in these groups were further categorised into their corresponding biopsy techniques: (A) laser+pulling; (B) laser+flicking; (C) flicking only.

Participants/materials, setting, methods
Blastocysts which were at least fair graded (Gardner, 1999) were biopsied and vitrified (Cryotec, Japan). The number of biopsied cells ranged from 5 to 10 cells. All biopsied trophectoderm samples were subjected to Preimplantation Genetic Testing for Aneuploidy (PGT-A) with Next Generation Sequencing (NGS) (Ion Torrent, USA). Chromosomal mosaicism analysis was done using ReproSeq Mosaic PGS w1.1 workflow. Trophectoderm biopsied sample which were tested to have 20% to 80% aneuploid cells were reported as mosaic.

Main results and the role of chance
In Group 1, the mosaicism rates for biopsy technique A, B and C were 23.3% (104/446), 28.2% (58/206) and 37.5% (6/16) respectively. In Group 2, the mosaicism rates for biopsy technique A, B and C were 14.6% (7/48), 19.5% (23/118) and 27.3% (3/11) respectively. There were no significant differences (p &gt; 0.05) in mosaicism rates between all study groups and subgroups.

Limitations, reasons for caution
Although no statistical significance was found between trophectoderm biopsy techniques and the prevalence of mosaicism, there is a trend of an increase in mosaicism rate when the flicking technique was used. Therefore, further studies with a larger sample size should be undertaken.
Wider implications of the findings: Our study demonstrates a trend in the decrease of mosaicism rate when laser pulses was used to loosen the cell junction of targeted cells. Hence, in place of the flicking method alone, laser pulses should be applied during trophectoderm biopsy if our findings are confirmed in a larger controlled study.

Trial registration number
Not applicable
P–575 Patients with recurrent implantation failures (RIF): chromosome abnormalities in the resulting embryos
Authors: Albanese C , Perruzza D , Tabanelli C , Sgargi S , Magli M C , Gianaroli L 
Published: August 2021

Abstract

Study question
Do RIF patients have the preimplantation genetic testing for aneuploidy (PGT-A) overcome their infertility condition?

Summary answer
PGT-A positively impact on implantation rate in RIF patients

What is known already
The most common definition of RIF is failure to achieve a pregnancy after three consecutive transfers of good quality embryos. This term possibly represents a heterogeneous category of infertile couples as the causes of repeated failures can be diverse. Especially intriguing is the case of patients with an age lower than 39 years for which the oocyte quality is expected not to be compromised by the well known age effect on female fertility. The chromosome analysis of the resulting embryos has been proposed as a valid method to improve implantation in the great majority of RIF patients

Study design, size, duration
This retrospective study included 49 patients with at least three previous consecutive implantation failures, which underwent PGT-A from January 2016 to April 2020. Both partners had a normal karyotype. Only patients with a female age below 39 years were included, who presented with a normal uterine cavity. Couples with a severe male factor were excluded. Single frozen blastocysts were transferred according to chromosomal results

Participants/materials, setting, methods
Maternal age was 35.5 ± 3.1 years. All blastocysts were vitrified after trophectoderm biopsy. Whole genome amplification and array comparative genomic hybridization were performed on biopsies. Only euploid embryos were transferred. The primary outcome was the live-birth delivery rate after the first transfer

Main results and the role of chance
Before starting a PGT-A cycle, these patients underwent 213 embryo transfers with 251 embryos replaced. A total of 264 blastocysts were analyzed, 140 of which were aneuploid (53%). Monosomy or trisomy was reported in 67 of the diagnosed samples (67/140, 48%) whereas the remaining 73 carried complex aneuploidies (73/140, 52%). The remaining 124 blastocysts (47%) were diagnosed as euploid. All patients performed an embryo transfer resulting in 28 clinical pregnancies (57%). There were 5 spontaneous abortions and the live-birth delivery rate per patient was 47%

Limitations, reasons for caution
This study suffers from the weakness related to retrospectivity. In addition, as euploid embryos are still cryopreserved, the delivery rate could change at completion of the cycles
Wider implications of the findings: A RIF condition can be attributed, at least in a good proportion of cases, to the generation of high percentages of aneuploid embryos. In this case, the transfer of euploid blastocysts has high chances to classify this category of RIF patients has having an embryonic cause of infertilit.

Trial registration number
Not applicable
P–578 Human pappiloma virus 16,18 genome methylation patterns in subfertile women
Authors: Mastora E , Zikopoulos A , Galani A , Georgiou I , Zikopoulos K 
Published: August 2021

Abstract

Study question
A comparison between L1 gene and LCR region methylation status of HPV16 and HPV18 viruses in subfertile women, investigating HPV methylation pattern in cervical cancer and asymptomatic HPV infection.

Summary answer
CpG methylation was more frequent in L1 gene compared to LCR in both HPV types. Methylation levels were associated with the grade of cervical dysplasia.

What is known already
HPV infection is a common sexually transmitted disease, related to genital warts and cancer. DNA methylation as a dynamic and strictly controlled process can be involved in numerous cellular processes, cell differentiation, gene expression regulation and genome reprogramming. Human pappiloma virus genome epigenetic alterations may play a key role in HPV life cycle as well as in the oncogenic process in general. However, whether the prevalence of high risk HPV is correlated with female infertility, has yet to be elucidated.

Study design, size, duration
From January 2015 to December 2019, about 2505 infertile couples were referred to the Human Reproduction Unit of Ioannina University Hospital. A total of 212 clinical and laboratory data from female partners were included in the study.

Participants/materials, setting, methods
Cervical smears were studied for HPV DNA methylation. CpG methylation was compared among L1 gene and LCR region in both HPV types. A bisulfite modification assay followed by DNA amplification and sequencing was performed to analyse HPV16 and HPV18 genome.

Main results and the role of chance
In HPV16 types, L1 gene and promoter region indicated high methylation levels in cervical cancer cases. LCR regions methylation levels ranged from 0,5% to 24,2% in asymptomatic HPV16 infection or cervical intraepithelial neoplasia and cervical cancer, respectively. As for L1 gene, the differences between asymptomatic HPV16 infection and cervical cancer cases were statistically significant (P = 0.003). In HPV18 types, L1 gene was methylated in cervical intraepithelial neoplasia and cervical cancer cases. Promoter region methylation levels were high in cervical cancer cases while LCR region methylation levels were low.

Limitations, reasons for caution
Main limitation is the relatively small size of the collected samples.
Wider implications of the findings: HPV genome investigation, as for methylation status, may lead to better understanding and earlier diagnostics of cervical pathology in infertile population. These observations point out the importance of fertility preservation in women at high risk for cervical neoplasia.

Trial registration number
Not applicable
P–581 The day of blastocyst biopsy and the chromosomal constitution. Evaluation of 5125 embryos by Next Generation Sequency (NGS)
Authors: Villanuev. Zúñiga P E , Huayhua J , Noriega-Hoces L , Llerena G , Noriega-Portella J , Noriega-Portella L , Guzman L 
Published: August 2021

Abstract

Study question
Is there a relationship between the day of blastocyst biopsy and the results NGS analysis?

Summary answer
Embryos biopsied on day 6 or 7 are associated with the increased probability of being an aneuploidy embryo and less likely to be mosaic embryo.

What is known already
There is controversy about whether an embryo that reaches the blastocyst stage on day 5 has a higher chance of being euploid than embryos which are biopsied later. In our study, chromosome constitution was evaluated by next-generation sequencing (NGS)-based preimplantation genetic testing for aneuploidy (PGT-A) and confounding factors were eliminated.

Study design, size, duration
Data was collected retrospectively from June 2016 to January 2020

Participants/materials, setting, methods
In total, 5125 blastocyst (day 5=2914, day 6 N = 2154 and day7 N = 57), generated from 1318 cycles were analysed with PGT-A. The chromosome constitution for each embryo was classified as euploid, aneuploid and mosaic. A multilevel model was made and associations betwwen variables by logistic regression were adjusted according to maternal age, SART blastocyst grade, fertilization method, biopsy operator and blastocyst stage.

Main results and the role of chance
The mean maternal age was 36.2 ± 4.2. Euploid rate was 62.1% and 37.9% (day 5 and day 6–7 respectively), aneuploidy rate was 47.0% and 53.0% (day 5 and day 6–7, respectively), mosaicism rate was 59.6% and 40.4% (day 5 and day 6–7, respectively) (p &lt; 0.001).
Embryos biopsied on day 6–7 have a significantly lower probability to be euploid and mosaicism than embryos biopsied on day 5 ((OR = 0.76 [0.68–0.86]); (OR = 0.84 (0.73 – 0.96) respectively) (p &lt; 0.001). On the contrary, embryos biopsy on day 5 were significantly more likely to be euploid than day 6–7 (OR = 1.63[1.42–1.86]) (p &lt; 0.001).

Limitations, reasons for caution
The results observed in this study should be confirmed using a larger number of samples. For the NGS analysis, a chromosome with a variation between 20 to 80% was considered mosaic.
Wider implications of the findings: The present study revealed that embryos that reach blastocyst classified as full to hatched on day 5 are more like to be euploid compared to slow growing embryos.

Trial registration number
non-clinical trials
P–589 The effects of vanillic acid on production of testosterone and function of insulin in adipose and ovarian tissues in rat model of polycyctic ovary syndrome
Authors: Nejabati H R , Shahnazi V , Ghaffar. Novin M , Nouri M 
Published: August 2021

Abstract

Study question
What is the possible therapeutic effects of vanillic acid (VA) on the treatment of Polycystic Ovary Syndrome (PCOS)?

Summary answer
Vanillic acid successfully alleviated metabolic and endocrine abnormalities in adipose and ovarian tissues in a rat model of PCOS.

What is known already
Polycystic Ovary Syndrome (PCOS), as a common endocrine disorder, is accompanied by hyperandrogenism, insulin resistance, ovulation problems, and infertility. In this study, the effects of vanillic acid (VA) on metabolic and endocrine abnormalities were evaluated in the adipose and ovarian tissues of a letrozole-induced rat model of PCOS.

Study design, size, duration
In this study, a letrozole-induced rat model was established and then the experimental groups were treated by VA and MET through oral gavage for one month. Before and after the treatment with drugs, the vaginal smear was carried out . Ovarian and adipose tissues were collected and frozen at − 80 for further analysis following euthanasia of the rats.

Participants/materials, setting, methods
Thirty Six -week- old Wistar rats were divided into four groups (six rats per each group). Twenty-four rats were received letrozole for five weeks. Then, two experimental groups were treated by metformin (MET), and VA . Serum lipid profile, fasting glucose levels, and hormonal assays were done by corresponding commercial kits. The phosphorylation of AMP-activated protein kinase (AMPK) was detected by western blotting and the expression of studied genes were measured by Real-time PCR.

Main results and the role of chance
Our results showed that both VA and MET successfully reversed the abnormal estrous cycles of PCOS rats and reduced the serum testosterone levels and Steroid 17-alpha-hydroxylase/17,20 lyase (CYP17A1) gene expression. Furthermore, they improved Homeostatic Model Assessment for Insulin Resistance (HOMA-IR) and decreased the serum glucose and triglyceride levels, and gene expression of resisitin. The phosphorylation of AMPK was significantly decreased in adipose and ovarian tissues in PCOS group and both therapeutic factors successfully activated the AMPK in these tissues. However, VA had not such a significant effect in adipose tissue.

Limitations, reasons for caution
The main limitation of the current study was its design as a rat model, which may have limitation in the translation of knowledge to the clinics.
Wider implications of the findings: In summary, to the best of our knowledge, this study is the first study reported beneficial effects of VA on the treatment of PCOS. The findings of the current study shed light on an urgent need for discovering novel therapeutic pharmaceuticals regarding the treatment of the PCOS.

Trial registration number
0