bluuebunny/crossref_metadata_2025_split huggingface dataset:
P-291 Positive effects of inactivated blood serum in stabilizing the activity of antioxidants in embryo-free culture media
Authors: Ramirez-Dominguez L.B , Jimenez Medina I , Maldonado-Rosas I , Villar-Muñoz L.G , Saleh R , Agarwal A 
Published: June 2022

Abstract

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

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

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

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

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

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

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

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

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

Abstract

Study question
Which are the euploid results using niPGT?

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

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

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

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

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

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

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

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

Abstract

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

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

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

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

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

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

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

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

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

Abstract

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

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

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

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

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

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

Limitations, reasons for caution
Cell mortality in vitro and animal survival after cell injection were among the limitations of the study.

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

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

Abstract

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

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

What is known already
A number of factors have been reported as predictors for Misoprostol treatment failure, including assisted reproductive treatment, primigravity, and vaginal bleeding prior to treatment.
Increased rates of failure with medical management were also reported with delaying treatment after diagnosis.

Study design, size, duration
This was a retrospective cohort study.
Overall, 778 patients who underwent medical treatment with Misoprostol for missed abortion at our institute, between 07/2015 and 12/2020 were included.

Participants/materials, setting, methods
Inclusion criteria were either anembryonic gestation or embryonic death, as long as pregnancy age, as measured by transvaginal sonography , correlated with 12 weeks gestation or less.
Exclusion criteria included incomplete or septic abortions, multiple pregnancies, and cases that had either irregular period, missing data, or were lost to follow up.
The primary outcome was treatment failure, defined as any need for surgical intervention (elective or emergency Dilation & curettage, or operative hysteroscopy).

Main results and the role of chance
Five hundred and eighty two patients (74.9%) had successful medical treatment, while 196 (25.1%) required surgical intervention.
The gestational age based on menstrual dating and on ultrasound difference (in weeks) was 2.6±1.4 in the success group and 3.1±1.6 in the failure group (p &lt; 0.001).
On multivariant regression analysis, gestational age based on menstrual dating and on ultrasound difference was found to be independently correlated with treatment failure (aOR=1.24, CI 95% (1.01-1.51), p = 0.03).

Limitations, reasons for caution
We did not analyze long term consequences, such as the amount of bleeding, and time to future fertility.
Moreover, patients who were treated at our institute may have been diagnosed and treated for treatment failure in neighboring hospitals and hence lost to follow-up.

Wider implications of the findings
The current study has shown that the difference between gestational age based on menstrual dating and on ultrasound is directly corelated with medical treatment failure.
This data should be taken into account when counseling patients regarding the optimal treatment for first trimester missed abortion.

Trial registration number
not applicable
P-436 EndoTime: Non-categorical timing estimates for luteal endometrium
Authors: Mitchell A , Lipecki J , Muter J , Lucas E , Makwana K , Fishwick K , Odendaal J , Hawkes A , Vrljicak P , Brosens J , Ott S 
Published: June 2022

Abstract

Study question
Can expression measurements of a small panel of genes be used to develop a continuous, non-categorical model for the improvement of endometrial biopsy timing accuracy?

Summary answer
Measuring expression levels of six genes (IL2RB, IGFBP1, CXCL14, DPP4, GPX3, and SLC15A2) is sufficient to obtain and assess substantially more accurate timing estimates.

What is known already
Commercially available endometrial timing approaches based on gene expression require much larger gene sets and use a categorical approach that classifies samples as pre-receptive, receptive, or post-receptive.

Study design, size, duration
Gene expression was measured by RT-qPCR in different sample sets, comprising a total of 664 endometrial biopsies obtained 4 to 12 days after a self-reported positive home ovulation test. A further 36 endometrial samples were profiled by RT-qPCR as well as RNA-sequencing.

Participants/materials, setting, methods
A computational procedure, named ‘EndoTime’, was established that models the temporal profile of each gene and estimates the timing of each sample. Iterating these steps, temporal profiles are gradually refined as sample timings are being updated, and confidence in timing estimates is increased. After convergence, the method reports updated timing estimates for each sample while preserving the overall distribution of time points.

Main results and the role of chance
The Wilcoxon rank-sum test was used to confirm that ordering samples by EndoTime estimates yields sharper temporal expression profiles for held-out genes (not used when determining sample timings) than ordering the same expression values by patient-reported times (GPX3 : p &lt; 0.005; CXCL14 : p &lt; 2.7e-6; DPP4 : p &lt; 3.7e-13). Pearson correlation between EndoTime estimates for the same sample set but based on RT-qPCR or RNA-sequencing data showed high degree of congruency between the two (p = 8.6e-10, R-squared = 0.687). Estimated timings did not differ significantly between control subjects and patients with recurrent pregnancy loss or recurrent implantation failure (p &gt; 0.05).

Limitations, reasons for caution
Timing estimates are informed by glandular gene expression and will only represent the temporal state of other endometrial cell types if in synchrony with the epithelium. Additionally, methods that estimate the day of ovulation are still required as these data are essential inputs in our method.

Wider implications of the findings
Our method is the first to assay the temporal state of endometrial samples on a continuous domain, enabling accurate temporal profiling of any gene in luteal phase biopsies for a wide range of research applications and, potentially, clinical use. It is freely available, open-source software including supporting data sets.

Trial registration number
Not applicable
P-561 Long-read amplicon guided haplotype imputation enabling comprehensive preimplantation genetic testing in families with de novo pathogenic variants
Authors: Ayeb Y , Tsuiko O , Jatsenko T , Allemeersch J , Melotte C , Ding J , Denayer E , Legius E , Brems H , Vermeesch J , Dimitriadou E 
Published: June 2022

Abstract

Study question
Can we use comprehensive genome-wide haplotyping PGT for couples carrying de novo mutations, given the absence of phasing references to establish the disease-associated haplotype?

Summary answer
Targeted amplicon long-read sequencing represents a valuable approach for PGT workup that leverages comprehensive PGT application in families with de novo mutations.

What is known already
Preimplantation genetic testing (PGT) aims to select embryos devoid of inherited pathogenic variants. Current comprehensive genome-wide haplotyping PGT methods cannot be applied for couples where one of the partners carries a de novo mutation, as absence of affected close relatives restricts variant phasing to establish the disease-associated haplotype. For such families targeted approaches are currently used instead, thus missing genome-wide embryo analysis.

Study design, size, duration
To overcome the forementioned limitation, we developed a long-read amplicon guided haplotype imputation method. From January 2017 till June 2021 thirty-two couples were enrolled into the PGT program at the Centre for Human Genetics, UZ Leuven, in the context of a de novo variant causing a known monogenic disorder in one of the two partners.

Participants/materials, setting, methods
Genomic DNA from the partner carrying the variant (proband) and his/her parents was used for trio analysis via long-range PCR and long-read amplicon sequencing using the PacBio RSII and/or Oxford Nanopore platforms. Targeted haplotype phasing was then performed to impute the disease-associated allele.

Main results and the role of chance
The parental origin of the mutant allele was identified in 20 patients, resulting in the current 62.5% success rate. Most de novo mutations occurred on the paternal allele. In the remaining families, the region of interest either had no SNPs or had insufficient number of informative SNPs linked to mutation. From the successfully analyzed couples, eight have proceeded to PGT cycle and so far, three disease-free children have been born.

Limitations, reasons for caution
A costly family-specific work-up is required, allowing nevertheless the implementation of a generic genome-wide method at the embryo analysis stage. The sequencing cost itself is anticipated to decrease in time. To avoid non-informative results, the analysis of multiple amlpicons is recommended.

Wider implications of the findings
Current practices for the handling of couples with de novo variants are mostly targeted, require the combination of direct and indirect approaches and often require the multiple biopsies. Targeted amplicon long-read sequencing represents a valuable approach for PGT workup that leverages comprehensive PGT application in families with de novo mutations.

Trial registration number
not applicable
P-580 Cumulative live birth rate of oocyte in-vitro maturation with a pre-maturation step in women with polycystic ovary syndrome or high antral follicle count
Authors: Pham H , Le A , Nguyen A , Ha U , Nguyen T , Pham T , Ho T , Vuong L 
Published: June 2022

Abstract

Study question
What is the cumulative live birth rate (CLBR) at 24 months of women undergoing oocyte in vitro maturation (IVM) a with pre-maturation step (CAPA-IVM)?

Summary answer
The CLBR at 24 months with CAPA-IVM was 38.2%.

What is known already
IVM with a pre-maturation step, known as capacitation IVM (CAPA-IVM) improves the competence of oocytes matured in-vitro by sustaining meiotic arrest to allow the synchronization of cytoplasmic and genetic maturation of the oocytes. Results from a randomized controlled trial showed that CAPA-IVM resulted in a live birth rate after the first transfer of 35.2% which was non-inferior to IVF. There is a lack of data on cumulative live birth following CAPA-IVM.

Study design, size, duration
A multi-center, retrospective study, performed at IVFMD, My Duc Hospital and IVFMD Phu Nhuan, My Duc Phu Nhuan Hospital from 1 January 2017 to 31 December 2019.

Participants/materials, setting, methods
All women with polycystic ovary syndrome (PCOS) or high antral follicle count (AFC) treated with a CAPA-IVM cycle were recruited to the study. All embryos were frozen at day 3. Cumulative live birth was difined as at least one live birth resulting from initiated CAPA-IVM cycle. If the women did not return for embryo transfer, outcomes were followed up until 24 months from aspiration day. Logistic regression was performed to assess which factors predicted CLBR.

Main results and the role of chance
Between 1 January 2019 and 31 December 2019, there were 374 eligible women included in the study, among them, 368 patients had embryos for transfer (98.4%) and six patients had no embryo for transfer (1.6%). A total of 496 frozen embryo transfer (FET) cycles from 368 patients were performed. Mean age and body max index (BMI) were 29.5±3.21 years and 22.3±3.09, respectively. The maturation rate was 63.2%. The median number of embryos was 4.0 [2.00; 6.00]. The cumulative clinical pregnancy rate was 60.4%, cumulative ongoing pregnancy rate was 51.6%. At 24 months after starting the treatment, the CLBR of CAPA-IVM was 38.2%. Multivariate analysis showed that patient age and number of metaphase II oocytes were the predictive factors for cumulative live birth after CAPA-IVM.

Limitations, reasons for caution
The study limitation derives from its retrospective nature. The generalizability of the study may be limited due to the single ethnicity group.

Wider implications of the findings
In patients with PCOS or high AFC, CAPA-IVM was viable and resulted in acceptable CLBR. CAPA-IVM may be the first-line treatment for women with PCOS or high AFC requiring assisted reproduction.

Trial registration number
not applicable