bluuebunny/crossref_metadata_2025 huggingface dataset:
P–537 What is the attitude of gamete donors towards expanded genetic testing
Authors: Skytte A B , Pacey A , Rothma. Herrmann J , Mocanu E , Burke C , Pennings G
Published: August 2021

Study question

What is the opinion of gamete donors on extended carrier screening in Denmark and in US?

Summary answer

This study showed that the overwhelming majority of the donors were very positive towards genetic testing in general and the expanded carrier screening.

What is known already

There is a lack of empirical studies on the experiences of and attitudes of donors towards expanded carrier screening (ECS) (Amor et al. 2018). Oocyte donors in a Spanish clinic were surprised by the information on testing and the possibility of being carriers (Abuli et al., 2016). After adequate genetic counselling before and after the test, the results of testing did not seem to have a meaningful emotional or psychological impact on the donors.

Study design, size, duration

A questionnaire was emailed to all active sperm donors in a sperm bank in Denmark and in a sperm bank in US.

Participants/materials, setting, methods

A total of 393 donors received the email of which 233 donors completed the questionnaire, 196 in Denmark (response rate of 60.7%) and 37 in the United States (response rate of 52.9%).

Main results and the role of chance

We systematically compared the US and DK donors and ID-release versus non-ID-release donors for all characteristics and items. ID-release donors with a partner significantly more informed their partner about their donor status than non-ID-release donors (90.5% vs. 72.4%, p 0.027). ID-release donors significantly more intended to tell their own children (57.5% vs. 21.2%, p 0.001). ID-release donors significantly more thought about their potential donor children (56.2% vs. 35.0%; p 0.002) and significantly more likely to want information on whether a pregnancy occurred from their donation (70.6% vs. 55.0%, p 0.017). In addition, they also significantly more wished to obtain information about the children conceived with their sperm: the number of children (64.0% vs. 50.0%, p 0.048), their gender (40% vs. 26.2%, p 0.042), and their health (59.3% vs. 42.5%, p 0.018). The ID-release donors were much less likely than the other type not to want any information (19.3% vs. 32.5%, p 0.034). This general attitude is extended to the wish to be informed if a child with a hereditary disease is born after using their sperm. The ID-release donors were more likely to desire contact if their genetic child was diagnosed with a genetic disease (73.9% vs. 55.0%, p 0.003).

Limitations, reasons for caution

The main element that makes the study unique (i.e., the choice between ID-release and non-ID-release) also renders the generalization of the findings more difficult. Wider implications of the findings: A better understanding of the attitude among gamete doners will possibly guide future legislation and ensure the safety and welfare of the donors too.

Trial registration number

none
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

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

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

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

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 > 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

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

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

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 < 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 < 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 < 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

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
P–598 Further evidence for a functional hormonal adrenal-ovarian axis affecting female infertility
Authors: Benor A , Molinari E , Barad D H , Gleicher N
Published: August 2021

Study question

How does here presented case offer further evidence for existence of a functional hormonal adrenal-ovarian axis?

Summary answer

This is the first case of iatrogenic Cushing syndrome leading to severe adrenal and ovarian insufficiency, as evidenced by undetectable estrogen and low androgen levels.

What is known already

Animal models and human data have convincingly demonstrated that hypo-androgenism affects follicle recruitment and growth, especially at small growing follicle stages, in most severe cases even mimicking primary ovarian insufficiency (POI). In milder forms, hypoandrogenism reduces follicle number, egg numbers as well as egg quality, unless reconstituted in timely fashion before IVF cycle start.

Study design, size, duration

We here report a 34-year-old G1P1, who presented for a second opinion with a diagnosis of secondary “unexplained” infertility after two IVF cycles at another fertility center.

Participants/materials, setting, methods

Since our center considers a diagnosis of “unexplained” infertility as subjective, the patient underwent a thorough diagnostic evaluation. She was using oral contraceptive pills for one week at the time her laboratory results were drawn. Main results and the role of chance: Her free (FT) and total testosterone (TT) (0.4 pg/ml and 5.0 ng/dL, respectively), DHEA and DHEAS (103.0 ng/dL and 92.0 µg/dL, respectively) were low and her estradiol was undetectable (<25 pg/mL), reflecting significant adrenal as well as ovarian suppression. Morning ACTH was undetectable at < 5 pg/mL but cortisol was abnormally elevated (17.7mcg/dL), leading to diagnoses of secondary adrenal insufficiency as well as secondary ovarian insufficiency (SOI) due to adrenal hypo-androgenism from lack of ACTH production. She, in addition, revealed a positive ANA titer (1:160). Because of eczema, she for over a year had been on a super-potent topical steroid ointment. Upon termination of this steroid, adrenal as well as ovarian function, as evidenced by her hormonal values, normalized.

Limitations, reasons for caution

This is the first case in the literature where iatrogenic-induced insufficiency of adrenal androgen production resulted in secondary ovarian insufficiency (SOI), characterized by undetectable estradiol, reversible by withdrawal of topical steroid treatment.

Wider implications of the findings: This case offers further evidence that the traditional hypothalamic-pituitary-ovarian axis (HPAA) extends downstream to ovaries (HPAOA), reaffirming the ability of adrenals to control ovarian function.

Trial registration number

n/a
P–600 Ovarian function following intraovarian injection of autologous platelet rich plasma (APRP) in women with low functional ovarian reserve
Authors: Barad D , Darmon S K , Benor A , Gleicher N
Published: August 2021

Study question

Does exposure of ovaries to autologous growth factors in platelet rich plasma (APRP) affect the pituitary ovarian axis?

Summary answer

Within 60 days after injection, growing follicle numbers and estrogen levels increased, though FSH did not change, with effects most pronounced in still menstruating women.

What is known already

APRP is extracted from a patient’s autologous blood and delivers growth factors. It is widely used in several medical specialties and has in infertility practice been reported to increase folicle/egg numbers if injected into ovaries and improve endometrial thickness/implantation if used for perfusion of the endometrium.

Study design, size, duration

Prospective observational cohort study of women with low functional ovarian reserve, followed for 60 days after subcortical injection of ovaries.

Participants/materials, setting, methods

44 women with prior poor response to ovulation induction, FSH > 12 mIU/mL and AMH < 1.0 ng/mL. APRP was prepared using Regen Lab PRP Kit which is approved by the US-FDA. 1.0–1.5 ml of PRP was injected into the cortex of each ovary divided among 7 to 10 injection sites. Participants were followed every three days with monitoring for estradiol, FSH and follicle growth for the first two weeks after PRP and then weekly.

Main results and the role of chance

21/43 patients still regularly menstruated (subgroup A, age 43.9 ± 5.1 years); 23/43 (subgroup B, age 42.6 ± 6.2 years) were amenorrheic for a median of 6 months. In A, AMH, FSH and estradiol were 0.18 ± 0.20 ng/mL, 37.5 ± 47.6 mIU/mL, and 100.2 ± 73.4 pg/mL, while in B they were 0.06 ± 0.11 ng/mL, 73.0 ± 44.8 mIU/mL and 66.7 ± 57.6 pg/mL. Following APRP, A-patients demonstrated increased estradiol to 211 ± 193.7 pg/mL (P = 0.029) while B-patients only demonstrated a trend to 98.1 ± 86.5 (P = 0.09). Among A patients, 14/21 (66.7%) entered IVF cycles and 5/21 (23.8%) reached retrieval. So-far 1 patients established an ongoing clinical pregnancy. Among B patients 8/23 (34.8%) entered IVF cycles and only 2/23 (8.7%) reached retrieval and none achieved pregnancy.

Limitations, reasons for caution

This observational study was only carried out to estimate possible effects of APRP treatments. Based on these observations, we are now conducting a randomized controlled trial, limited to cycling women under age 45 years [registration # NCT04278313].

Wider implications of the findings: PRP appears to have limited ability to affect ovarian reserve of older, and especially amenorrheic women. It may, however, exert more favorable effects on still menstruating women. Promotion of APRP treatment as “ovarian rejuvenation,” however, appears to be an inappropriate choice of words.

Trial registration number

N/A
P–645 Abnormal thyroid hormone levels are associated with biochemical primary ovarian insufficiency and low ovarian reserve: a cross-sectional study
Authors: Feng C
Published: August 2021

Study question

To address whether there was a correlation between thyroid parameters and primary ovarian insufficiency.

Summary answer

Thyroid dysfunction was related with bPOI and LOR.

What is known already

In our daily clinical work, there seems to be a link between thyroid disease and POI. A recent study reported that subclinical hypothyroidism was associated with lower ovarian reserve during later reproductive age. Animal studies demonstrated that thyroid hormones played important roles in ovarian functions, and both maternal hypothyroidism and hyperthyroidism were related with reduced primordial, primary and secondary follicle number in rats. However, others reported that thyroid autoimmunity (TAI) and hypothyroidism were not associated with DOR. In all, accumulative animal and epidemiological studies indicated the connection between thyroid function and ovarian reserve, but the results were still inconsistent.

Study design, size, duration

This is a cross-sectional study with consecutive women performed ovarian reserve assessment and thyroid test in the Second Hospital of Zhejiang University School of Medicine from April 2016 to March 2019. A total of 2109 women were included, with 111 with bPOI and 1771 without bPOI. To exclude the influence of age the participants were categorized into low ovarian reserve (LOR) and non-LOR groups based on age-specific AMH, including 78 LOR and 2031 non-LOR.

Participants/materials, setting, methods

At the outpatient, a doctor carried out an interview, recorded age, body mass index (BMI), past history, and current treatment, and made a diagnosis. Serum AMH, FSH, TPOAb, TgAb, Tg, TT3, FT3, TT4, FT4, and TSH levels were measured with electrochemiluminescence method.

Main results and the role of chance

TT3, FT3, FT4 was significantly positively correlated with serum AMH level. Further logistic regression analysis found that abnormal TT3, FT3 and TT4 levels were related to increased risks of bPOI and LOR. Chi-square analysis also proved that the incidence of abnormal TT3, FT3 and TT4 increased significantly in women with bPOI or LOR. The incidence of bPOI and LOR increased significantly in women with 2 or 3 abnormal thyroid hormones. The above analyses demonstrate in multiple aspects that thyroid dysfunction is related with decreased ovarian reserve.

Limitations, reasons for caution

Since this is a retrospective cross-sectional study, we only got the correlation between factors, and we could not achieve causal relationship. Further prospective cohort or randomized controlled trial (RCT) studies are required to make the results more robust.

Wider implications of the findings: The present study demonstrated that thyroid dysfunction was related with bPOI and LOR. It might be thyroid hormones, not TSH or thyroid antibodies, played the major role in ovarian reserve impairment. The treatment of euthyroxine may improve the ovarian reserve function.

Trial registration number

Not applicable
P–647 How do migraine attacks change during puberty?
Authors: Boettcher B , Kyprianou A , Wildt L , Lechner C , Kößler M , Neururer S , Bettina T , Matthias B , Rostasy K , Rauchenzauner M
Published: August 2021

Study question

How do the stage of puberty and the menstrual cycle influence characteristics of migraine?

Summary answer

During puberty, the frequency of migraine attacks increases, especially during the follicular phase. The pattern of migraine changes to a typical adult pattern of migraine.

What is known already

Up to puberty, headaches are as common in girls as in boys. After the onset of puberty migraine is more prevalent in adolescent girls suggesting an association with sex hormones. Attacks of menstrual migraine are characterized by a longer duration, tend to be more severe, and are less responsive to acute medication compared to migraine attacks which are independent from the menstrual cycle phase.

Study design, size, duration

For this prospective cohort study 47 girls were recruited from two Departments of Pediatrics and Adolescent Medicine between 01/2016 and 12/2018.

Participants/materials, setting, methods

Girls between 7 and 18 years old, diagnosed with migraine without aura according to the “International Classification of Headache Disorders II” diagnostic criteria, took part. Three groups (pre-, peri-, and postpubertal) were formed according to the Tanner stage and the onset of a regular menstruation. Girls kept a daily headache and menstrual cycle diary over 8 weeks. Ovulatory cycles were analyzed by weekly progesterone saliva tests.

Main results and the role of chance

Three groups according to Tanner stage and onset of regular menstruation were compared: pre- (n = 16), peri- (n = 19) and post-pubertal (n = 12) girls. A significant difference in migraine frequency was found between pre- and post- pubertal girls (p = 0.005). Headache characteristics did not differ significantly between the three groups. Interestingly, a higher frequency of attacks in follicular phase occurred compared to luteal phase (p = 0.030).

Limitations, reasons for caution

Repeated blood sampling would have been a more reliable technique compared to saliva assays. The sample size is small.

Wider implications of the findings: During puberty, the number of migraine attacks but not the specific headache characteristics changes in adolescent girls which should be taken into consideration regarding the management of these patients.

Trial registration number

AN2013–0027
P–658 Lovastatin promotes the expression of LDL receptor and enhances E2 production in the cumulus cells
Authors: Kim Y J , Choi K H , Kang K Y , Park E A , Kim Y S , Kim M J , Kim H O , Koong M K , Kim Y S , Yoon T K , Ko J J , Lee J H
Published: August 2021

Study question

Lovastatin enhanced E2 productive ratios in the cumulus cells through promoted expression of Low-density lipoprotein receptor (LDLR).

Summary answer

Lovastatin up-regulated gene expression of LDLR in the CCs. And the high expression of LDLR promoted E2 productive ratios from CCs.

What is known already

We already reported that the up-regulation of LDLR correlated with clinical pregnancy. Therefore, we found lovastatin as an up-regulator of LDLR expression of clinical pregnancy.

Study design, size, duration

This is an expended study of LDLR to enhance steroidogenesis regarding the effect of lovastatin in the CCs. The collection of human cumulus cells was approved by the Institutional Research and Ethical Committees of CHA University (approval number: 1044308–201611-BR–027–04) from January to December 2019. The CCs were collected from 12 patients with normal ovarian response after oocyte denudation for ICSI.

Participants/materials, setting, methods

We studied whether lovastatin has up-regulated LDLR expression in human CCs. Cumulus cells were collected from patients with young (∼ 36) and old aged patients (37 ∼). After culturing human CCs, they were treated lovastatin for one day. The concentration of E2 in culture medium was measured using Chemiluminescence immunoassay. The mRNA isolated from CCs was analyzed gene expression level through real time-PCR.

Main results and the role of chance

The concentration of E2 was significantly increased in the culture medium treated with lovastatin. The CCs treated with lovastatin increased the expression of LDLR and StAR which are components of the steroidogenesis pathway.

Limitations, reasons for caution

We have found that the role of lovastatin promotes the E2 production by increasing the ldlr gene of CCs. Therefore, further investigations aimed at lovastatin effect on human oocytes embryo whether enhanced quality of oocytes or not.

Wider implications of the findings: Previous data show that high activation of LDLR and StAR was associated with embryo quality and clinical pregnancy in infertile women. Our data suggest that lovastatin is stimulated LDLR expression to enhanced pregnancy ratios of IVF patients.

Trial registration number

none
P–791 Comparison of the live birth rate after single embryo transfer in fresh versus frozen cycle: no evidence to support routine freeze-all strategy
Authors: Toikkanen R , Terho A , Pelkonen S , Martikainen H
Published: August 2021

Study question

Is the treatment outcome compromised after superovulation for fresh IVF/ICSI in comparison to frozen cycle with spontaneous ovulation and luteal support with progesterone?

Summary answer

Live birth rate (LBR) is dependent on embryo quality both in the fresh and frozen cycles with no sign of harmful effect of the superovulation.

What is known already

Freeze-all strategy has been advocated in recent years based on the assumption that luteal phase after superovulation is not optimal for embryo implantation. The effects of variable hormonal treatments, given in association with ART, on the endometrium, are still largely unknown. Therefore, more data is needed in order to optimize the treatment policies.

Study design, size, duration

This is an observational retrospective single-center cohort study. Data were collected from Oulu University Hospital’s ART-database including a total of 5647 single embryo transfer cycles from years 1995–2020. Patients stimulated with long agonist protocol for IVF/ICSI and day 2–3 transfer were included. Frozen embryo transfer was performed in a natural cycle with an ovulation test used for timing of transfer. Luteal support with progesterone was given for two weeks in all cycles.

Participants/materials, setting, methods

There were 3053 IVF/ICSI fresh cycles (2237 top and 816 N-top) and 2594 frozen cycles (806 top and 1788 N-top). The main outcome measure was LBR compared between fresh and frozen cycles when either a top or a N-top embryo was transferred. As a secondary outcome, clinical pregnancy rate was investigated. Data on the age and body mass index (BMI) of the patients was available. Student’s T-test was used to compare continuous variables.

Main results and the role of chance

The groups did not differ regarding the age and BMI of the patients. After the transfer of a top quality embryo the clinical pregnancy rate (35.4 vs. 30.8%; p < 0.05) and LBR (29.4 vs. 25.5%; p not significant) was slightly higher in the fresh cycle. After the transfer of a N-top quality embryo the clinical pregnancy rate (19.9 vs. 19.4%) and LBR (15.9 vs. 15.4%) were similar both in the fresh and frozen cycles.

Limitations, reasons for caution

This study only represents cleavage stage embryo transfers, and all FETs were performed in a natural cycle. In a retrospective study there may also be residual confounding that could not be excluded.

Wider implications of the findings: This study provides further evidence that treatment outcome regarding LBR is not affected by superovulation therapy. Hence, the use of freeze-all strategy is warranted only in cases with a risk of ovarian hyperstimulation syndrome.

Trial registration number

Not applicable
P–794 Prevalence of positivity for SARS-CoV–2 RNA in follicular fluid in infertile patients
Authors: Porcu E , Cipriani L , Dirodi M , Calza N , Ciotti P M , Tranquillo M L , Notarangelo L , Zuffa S , Labriola F S , Vocale C , Damiano G
Published: August 2021

Study question

Is Sars-Cov–2 present in the follicular fluid of infertile patients?

Summary answer

In the experience of the Infertility and IVF Unit, University Of Bologna, Italy, RNA of SARS-Cov–2 was not detected in the follicular fluid.

What is known already

Data on the risk of virus presence in reproductive cells and transmissibility in IVF procedures are very limited. In literature only one study reports the detection of SARS-Cov–2 viral RNA in oocytes of PCR positive women. Research of RNA in follicular fluid could be a marker able to indicate whether to continue IVF treatments in the case of swab-positive patients.

Study design, size, duration

Prospective study performed at Infertility and IVF Unit, Sant’Orsola University Hospital, University of Bologna, Italy, from March 2020 to January 2021. 451 IVF cycles were performed on 902 patients. In addition 59 cycles of oocyte cryopreservation were also performed to fertility preservation in oncological patients. In all positive swab patients was analyzed the follicular fluid for RNA virus detection.

Participants/materials, setting, methods

961 patients underwent telephone triage before going to the IVF Center to identify subjects with suspected or confirmed infection. Body temperature was measured on all patients before entering the IVF Center.

All patients were subjected to real-time analysis (RT PCR) of pharyngeal swab samples 48 hours before transvaginal ultrasound-guided oocyte retrieval.

In case of positive swab, PCR was performed on follicular fluid.

Main results and the role of chance

In our population of infertile patients the incidence of SARS COV–2 infection positivity was 0.4% (4/961).

No IVF treatments were suspended. The oocytes of the 4 women with positive swab were cryopreserved using closed devices stored in a special dedicated cryogenic container. No viral RNA was detected in the follicular fluid.

Limitations, reasons for caution

there are no limitations to the study.

Wider implications of the findings: The absence of SARS-COV–2 RNA in the follicular fluid is a reassuring result in the storage and future use of oocytes.

Trial registration number

Not applicable
P–797 A novel method for establishing human embryonic stem cells independent of feeder cells
Authors: Cai B
Published: August 2021

Study question

Is there a efficient establishing method of human embryonic stem cells directly from the human blastocysts independent of feeder cells?

Summary answer

We established a novel method of generating human embryonic stem cells directly from human blastocysts independent of feeder layer cells.

What is known already

Establishing embryonic stem cells lines mainly needed to coculture ICM clumps with feeder cells (like mouse or human fibroblasts) ,this brought in potential heterogeneous pollution.Although there had be some reports about generating human ESCs independent of feeder cells,but the efficiency was low and conditioned medium were unstable and also had the biological contamination.

Study design, size, duration

We used ten day5/6 donated human blastocysts from our reproductive center ,most of them were genetically diseased embryos with abnormal PGT diagnosis.After establishing ESCs procedure , all the cell lines were identified with pluripotency and differentiation potential tests.The success rate of system was calculated and compared with the conventional methods.

Participants/materials, setting, methods

In brief, ICM clumps were separated mechanically by using a micromanipulation system,and then transferred to a 30ul mTESR plus culture media drop pretreated with the geltrex (1:100 dilution) matrix and oxygen concentration was 5%. When cells attached and migrated,we also used laser to destroy the remaining trophoblast cells.About 10 days,the typical ES clone can be mechanically passaged and cells can be cultured in normal oxygen concentrations after passage 2. .

Main results and the role of chance

Using this method we had successfully established nine embryonic stem cell lines from donated human blastocysts ,the success rate was 90% (9/10). Each cell lines had passed the evaluation test of embryonic stem cell. When compared with the conventional feeder cells dependent method,our novol methods not only eliminated the pollution from heterogeneous cells,but also had higher success rate (90% vs 25%).

Limitations, reasons for caution

Due to the scarcity of donated human blastocysts, this experiment was a single-center experiment with small samples.

Wider implications of the findings: We speculated that the batch differences of culture dishes, matrix and culture medium might affect the establish efficiency , and how to carry out a high level of quality control work might be the key factor to keep the system stable.

Trial registration number

basic research
P2T2: Protein Panoramic annoTation Tool for the interpretation of protein coding genetic variants
Authors: DeVoe Elias , Oliver Gavin R , Zenka Roman , Blackburn Patrick R , Cousin Margot A , Boczek Nicole J , Kocher Jean-Pierre A , Urrutia Raul , Klee Eric W , Zimmermann Michael T
Published: August 2021

Motivation

Genomic data are prevalent, leading to frequent encounters with uninterpreted variants or mutations with unknown mechanisms of effect. Researchers must manually aggregate data from multiple sources and across related proteins, mentally translating effects between the genome and proteome, to attempt to understand mechanisms.

Materials and methods

P2T2 presents diverse data and annotation types in a unified protein-centric view, facilitating the interpretation of coding variants and hypothesis generation. Information from primary sequence, domain, motif, and structural levels are presented and also organized into the first Paralog Annotation Analysis across the human proteome.

Results

Our tool assists research efforts to interpret genomic variation by aggregating diverse, relevant, and proteome-wide information into a unified interactive web-based interface. Additionally, we provide a REST API enabling automated data queries, or repurposing data for other studies.

Conclusion

The unified protein-centric interface presented in P2T2 will help researchers interpret novel variants identified through next-generation sequencing. Code and server link available at github.com/GenomicInterpretation/p2t2.