bluuebunny/crossref_metadata_2025_split huggingface dataset:
P–148 Effect of additional laser assisted drilling (LAD) during trophectoderm (TE) biopsy on mosaicism rate
Authors: Tee Z Q , Chan C W , Lim A Y X , Lee C S S 
Published: August 2021

Abstract

Study question
Does applying additional LAD during TE biopsy cause higher mosaicism rate in blastocysts?

Summary answer
Applying LAD during TE biopsy to create additional zona opening will produce more mosaic blastocysts.

What is known already
In Alpha IVF, laser assisted hatching (LAH) was done on day3 after ICSI for all pre-implantation genetic testing for aneuploidy (PGT-A) cycles. TE biopsy techniques used were laser+pulling (L+P), laser+flicking (L+F) and flicking only (F). At the time of biopsy, an extra step of creating additinonal artificial opening by LAD may be required when (a) blastocyst has very little herniated cells; (b) inner cell mass (ICM) is at the hatching point or biopsy site. Our internal study showed that biopsy using different techniques (L+P, L+F and F) does not affect mosaicism rate.

Study design, size, duration
This prospective study was designed to evaluate the effect of additional LAD during TE biopsy on the mosaicism rate. This study was conducted between 11th March–19th August 2019. Four hundred forty-three (443) patients had undergone oocyte retrieval and blastocyst culture after intracytoplasmic injection (ICSI) was done. A total of 824 hatching blastocyst (BG5) and fully hatched blastocyst (BG6) with at least a Grade A or Grade B TE (Gardner’s grading) were included in this study.

Participants/materials, setting, methods
LAH was done on day3 post-ICSI while biopsy was done on day5 and/or day6 for PGT-A. Laser pulse length used during LAH and biopsy was fixed at 400ms. The biopsied blastocysts were classified into 3 groups: (A) BG6 (n = 79), (B) BG5 without additional LAD during biopsy (n = 713) and (C) BG5 with additional LAD (n = 32). The number of biopsied cells ranged from 5–10 cells. Biopsied cells were tested using Next Generation Sequencing (Ion Torrent, USA).

Main results and the role of chance
The mosaicism rates for Group A, B and C were 19.0% (15/79), 23.4% (167/713) and 39.5% (15/38) respectively. Mosaicism rates of Group A and B were comparable (p = 0.4807), whilst Group C had significant higher mosaicism rate compared to Group A and B (p = 0.0238 and p = 0.0319 respectively). The mean age of Group A, B and C were 31.1, 31.4 and 27.1 respectively. The mean age between these 3 groups were not statistically significant (A vs B, p = 0.0713; A vs C, p = 0.06727; and B vs C, p = 0.4408).

Limitations, reasons for caution
Additional LAD during TE biopsy maybe be a confounding variable which affects the mosaicism rate. Moreover, the increase in mosaicism rate could be due to other unknown factors. A larger sample size is needed to confirm the results.
Wider implications of the findings: Based on our study, additional LAD during TE biopsy is not recommended as this may increase mosaicism rate. Biopsy should be done when the blastocyst has more herniated cell or when the ICM leaves the hatching point/biopsy site.

Trial registration number
Not applicable
P–156 Automatic pronuclear detection based on deep learning technology has clinical utility
Authors: Takeda S , Fukunaga N , Sanami S , Tsuzuki Y , Kitasaka H , Takeda S , Watanabe H , Kida Y , Kondou F , Asada Y 
Published: August 2021

Abstract

Study question
Does the performance of an automatic pronuclear detection system based on deep learning technology have clinical utility?

Summary answer
Output results for 2PN detection using the automatic system powered by deep learning technology has clinical utility.

What is known already
In order to establish a more objective embryo evaluation system, we have been developing an automatic pronuclear detection system that utilizes deep learning technology based on Time- Lapse (TL) images. We have previously reported that the accuracy of pronuclei detection was improved by introducing an analysis method using 11 slices in the Z axis. In this study, we evaluated the potential clinical practicality of the automatic pronuclear detection system.

Study design, size, duration
Embryos clinically evaluated between May 2018 and December 2019 by embryologists were chosen for this study. We prepared for analysis TL videos of 995 embryos that had been evaluated as having 0, 1, 2, and 3PN.

Participants/materials, setting, methods
Part1:We compared the outputs of the automatic pronuclear detection system with these embryologists(three junior embryologists (1a), three intermediate embryologists (1b),and three senior embryologists (1c)) who had judged the pronuclei number from TL videos from 40 embryos each having 0,1,2,and 3PN.
Part2:The automatic pronuclear detection system determined the pronuclei number from the TL videos of 955 embryos scored as either 1,2,and 3PN,(different from those used in Part1),and the detection rate for 2PN was calculated.

Main results and the role of chance
Part1: The sensitivities for embryologist groups 1a),1b),1c) and the automatic pronuclear detection system were 80.0%,100%,100%,100% for 2PN, 60.0%,83.3%,86.7%,100% for 0PN, 46.7%,80.0%,86.7%,10.0% for 1PN, and 73.3%,96.7%,96.7%,10.0% for 3PN.
Part2: The precision for 2PN by the automatic pronuclear detection system was 99%.

Limitations, reasons for caution
In order to further improve the performance of the automatic pronuclear detection system, further adjustment of the algorithm and more training images will be utilised.
Wider implications of the findings: The detection of 2PN by the automatic pronuclear detection system was highly reliable, and the performance of the system was comparable to that of embryologists. These first results are reassuring and support the clinical use of the system as a further aid for embryologists, in routine laboratory practice.

Trial registration number
‘not applicable’
P–170 The secretomy of embryo sex
Authors: Orteiro M , Piccolomini M , Garcia C , Massaia I , Alvarenga A , Turco E. L , Duarte O , Yamakami L 
Published: August 2021

Abstract

Study question
Does the analysis of the metabolites of the embryonic culture medium can predict the sex of the embryo?

Summary answer
The presence and quantity of some metabolites in the culture medium can predict the sex of the human embryos.

What is known already
Advances in analytical techniques for metabolomics have brought the possibility of better tools for the characterization of molecules. Embryonic metabolism can be used as a good indicator of viability, regardless of the morphology of the blastocysts, since differences were observed in the metabolic activities between the days of embryo development and in the rates of live births.

Study design, size, duration
16 patients had their embryos biopsied between the months of January to July 2019 in a human reproduction laboratory. All cases had PGT-A indication and after the biopsy, the embryos were frozen. The culture medium samples were individually prepared for metabolites extraction according to the Bligh and Dyer protocol. Controlled ovarian stimulation and dose adjustments according to the response of each patient. The metabolomics analysis was performed by mass spectrometry.

Participants/materials, setting, methods
Follicular puncture were performed 35 hours after r-hCG. The eggs were kept in individual culture until the blastocyst stage. The blastocysts biopsy was performed (20). After the culture medium was sent to the 337 metabolites analysis by mass spectrometry. 14 molecules with the highest score on the PLS-Da was submitted to the ROC curves showing the power of metabolic analysis to predict the sex of euploid embryos. Besides, we performed the functional enrichment analysis.

Main results and the role of chance
After the genetic analysis by PGT-a, we obtain 20 euploid embryos, being 12 female embryos and 08 male embryos. Comparing the quantitative target metabolomic analysis of the 337 metabolites in the embryo culture medium, we observed the Asymmetric dimethylarginine, FAD, Malic Acid, Serotonin, increased in female embryos and Adenosine monophosphate, L-Alanine, L-Arginine, Cysteamine, DL-Dopa, Flavin Mononucleotide, Methionine sulfone, Nicotinic acid, L-Tyrosine, Uracil in male embryos. Through the ROC curve, we can verify AUC = 0.937. This result suggests that the metabolomic analysis of the culture medium is valid to be used as a complement of PGT-A to know embryo sex diagnostic. The functional enrichment analysis shows the Asymmetric dimethylarginine and Malic Sulfone metabolism as the principal function alter by female embryos.

Limitations, reasons for caution
Small number of samples
Wider implications of the findings: Further studies are needed to validate these findings for the diagnostic of sex embryos

Trial registration number
N/A
P–184 Assisted hatching does not improve live birth rates, a prospective double-blinded randomized study
Authors: Montalvo V , Masso J , Garcia-Faura A , Marques B , Lopez-Teijon M 
Published: August 2021

Abstract

Study question
Does Assisted hatching (AH) improve success rates when applied to frozen embryo transfers?

Summary answer
AH does not improve implantation, ongoing pregnancy or live birth rates when applied to thawed embryos.

What is known already
Vitrification has been proven to be the most efficient technique to preserve human embryos. However, vitrification has some consequences for the embryos, zona pellucida (ZP) hardening being one of them. Multiple studies suggest the need to apply laser Assisted hatching or ZP thinning to thawed embryos in order to improve success rates. Still, there is not enough evidence to ensure the utility of AH, and considering the great variation in design between studies more evidence is needed.

Study design, size, duration
Study performed from October 2019 and January 2020. Disregarding embryos with natural Hatching and PGT-A. Embryos that, immediately after thawing, were completely expanded (trophectoderm in contact with ZP) were also excluded from the study. We applied a randomization to choose in which embryos we had to perform AH. Neither the gynecologist nor the embryologist performing the embryo transfer knew whether the embryo had AH performed or not.

Participants/materials, setting, methods
353 frozen embryo transfers of one blastocist were considered for the study, 71 excluded for expansion after thawing, 65 excluded because of PGT-A, 103 in which we performed AH (AH+) and 114 without AH (AH-). In the AH+ group we performed laser-AH of 1/3 of the ZP, avoiding to damage the trophectoderm and performing the laser shots as far away to the ICM as possible. We used Chi-square testing to assess the effects of AH.

Main results and the role of chance
We assessed all relevant clinical data parameters. No statistical differences were found in egg age, maternal age, embryo quality, nor endometrial thickness between groups. Implantation and miscarriage rates were equivalent between AH+ group (40.9%; 20.5%) and AH- group (47.4%; 18.5%).
The main outcome of this study was live birth rates. No statistical differences were found between groups (AH-= 38.6%; AH + = 30.1%; p = 03221) proving that making it easier to get out of the ZP does not affect success rates.
Analyzing the data from the excluded embryos we found no improvement on live birth rates when embryos were expanded just after thawing (38.0%; p = 0.457). As expected, PGT-A embryos yielded higher live birth rates (52.3%; p &lt; 0,05)

Limitations, reasons for caution
Preliminary study with a small data set.
Wider implications of the findings: This study suggest that thawed embryos have the capacity to get out of the ZP regardless if AH was performed or not. Having no positive effects, AH seems to be unnecessary in this scenario.

Trial registration number
Not applicable
P–190 Can pregnancy be predicted according to mean infra-red attenuated total reflectance (MIR-ATR) spectrometry of incubation medium of embryos in IVF?: A proof of concept
Authors: Aslih N , Shalom-Paz E , Molenik D , Dekel B Z 
Published: August 2021

Abstract

Study question
Can MIR-ATR spectrometry of embryo incubation medium be used to predict embryo quality and IVF treatment results.

Summary answer
MIR-ATR spectrometry is able to distinguish between good and poor embryo quality and may improve the prediction of pregnancy based on better embryo selection.

What is known already
Infra Red (IR) spectra enable to determine if certain chemical functional components are present in a molecule. Attenuated total reflection (ATR) spectroscopy is a well established, simple and rapid technique for objective classification of biological samples. ATR spectrometry was investigated and confirmed to serve as an additional diagnostic tool in oral and gynecologic cancer. Currently, embryo quality is assessed based on morphokinetics parameters from Time-laps incubator and/or final embryo’s grading

Study design, size, duration
Culture media of 227 embryos on cleavage stage and blastocyst stage were collected and analysed between January 1st, 2018 and December 31st 2020.

Participants/materials, setting, methods
The incubation medium liquid of embryos cultured in Time-lapse incubator were collected after transferring the embryos to their final destination. Infra-Red (IR) absorbance spectra was measured. Nominal resolution was 4 cm–1, wavenumber range was from 600 cm–1 to 5000 cm–1 and each absorbance spectrum was normalized to the height of amide I band at ∼1650cm–1. Machine learning techniques utilized to build discrimination models for the absorbance data. Results were correlated with clinical and pregnancy results.

Main results and the role of chance
This preliminary study demonstrates that ATR spectrometry differs in media of : Day 3 embryos compared to day 5 embryos, top quality compared to poor quality embryos and implanting embryos comparing to non-implanting embryos. We found that MIR-ATR spectrometry might predict pregnancy in accuracy rates of 85%. MIR-ATR spectrometry of incubation medium can be used as an additional tool for selection of embryos for transfer.

Limitations, reasons for caution
Additional study and collection of larger number of culture Media is requested to validate MIR-ATR spectrometry as an additional tool in clinical set up in IVF units and laboratories.
Wider implications of the findings: MIR-ATR spectrometry can be used in the future as an additional tool for selection of embryos and prediction of cycles outcomes.

Trial registration number
NCT03317418
P–199 A case report to suggest that there must be other mutations than PATL2 or TUBB8 to cause oocyte maturation arrest
Authors: Molinari E , Yang M , Hu J , Zhang L , Albertini D F , Barad D H , Gleicher N 
Published: August 2021

Abstract

Study question
What causes our patient’s repeated almost complete oocyte maturation arrest (OMA)?

Summary answer
Since we did not detect PATL2 and TUBB8 mutations, both known to cause OMA, this case was likely caused by mutations in HUS1 and ITGB3

What is known already
OMA has been associated with loss-of-function in key genes, such as PATL2 and TUBB8. Such patients have, however, uniformly have been unable to conceive with IVF

Study design, size, duration
We here report the case of repeatedly presenting patient between 2009 until 2020 (age 30 at 1st and 41 at last visit).

Participants/materials, setting, methods
The couple underwent 7 IVF treatments under several ovarian stimulation protocols at different gonadotropin dosages and in different preparations to try to recruit mature eggs. She conceived in her 2nd IVF cycle in 2009 and delivered uneventfully in 2010. She then conceived spontaneously and delivered a healthy boy in 2014. The couple since then has been attempting another pregnancy. Remarkably, in all IVF cycles all eggs but one arrested at prophase.

Main results and the role of chance
The female demonstrates abnormally high ovarian reserve for age (AMH=5.9 ng/mL in 2019) (mean, 10.6 oocytes). In all cycles, all but one retrieved were immature. In vitro maturation rate for the GV oocytes was 28%. Resultant M2s, however, demonstrated morphological abnormalities, such as giant polar bodies. In vivo M2s, in contrast, were always morphologically unremarkable, and their fertilization rate was 85%. Embryo morphology deteriorated appreciatively with advancing age. Sanger sequencing for TUBB8 and PATL2 genes were unremarkable. Whole genome sequencing of her and her sister (who had no fertility problems) revealed mutations of genes belonging to the integrin family (ITGB3) and DNA repair checkpoint (HUS1), both of which could be determinants in the observed maturation arrest.

Limitations, reasons for caution
A functional study, coupled with imaging of the discarded material, will likely offer further information regarding the mechanisms leading to OMA in this female.
Wider implications of the findings: This case report represents a new phenotype of female infertility, characterized by almost complete maturation arrest which, however, still offers opportunity for pregnancy. Further isolation of underlying mutation(s) may offer additional insights about checkpoints required for the transition of prophase to metaphase in human oocytes.

Trial registration number
NA
P–219 mtDNA content in bovine cumulus cells does not predict oocytés developmental competence
Authors: Martíne. Moro Á , Lamas-Toranzo I , González-Brusi L , Pérez-Gómez A , Bermejo-Álvarez P 
Published: August 2021

Abstract

Study question
Does cumulus cell mtDNA content correlate with oocyte developmental potential in the bovine model?

Summary answer
The relative amount of mtDNA content did not vary significantly in oocytes showing different developmental outcomes following IVF

What is known already
Cumulus cells are closely connected to the oocyte through transzonal projections, serving essential metabolic functions during folliculogenesis. These oocyte-supporting cells are removed and discarded prior to ICSI, thereby constituting an interesting biological material on which to perform molecular analysis aimed to predict oocyte developmental competence. Previous studies have positively associated oocytés mtDNA content with developmental potential in both animal models and women. However, it remains debatable whether mtDNA content in cumulus cells could be used as a proxy to infer oocyte developmental potential.

Study design, size, duration
Bovine cumulus cells were allocated into three groups according to the developmental potential of the oocyte: 1) oocytes developing to blastocysts following IVF (Bl+Cl+), 2) oocytes cleaving following IVF but arresting their development prior to the blastocyst stage (Bl-Cl+), and 3) oocytes not cleaving following IVF (Bl-Cl-). Relative mtDNA content was analysed in 40 samples/group, each composed by the cumulus cells from one cumulus-oocyte complex (COC).

Participants/materials, setting, methods
Bovine cumulus-oocyte complexes were obtained from slaughtered cattle and individually matured in vitro (IVM). Following IVM, cumulus cells were removed by hyaluronidase treatment, pelleted, snap frozen in liquid nitrogen and stored at –80 ºC until analysis. Cumulus-free oocytes were fertilized and cultured in vitro individually and development was recorded for each oocyte. Relative mtDNA abundance was determined by qPCR, amplifying a mtDNA sequence (COX1) and a chromosomal sequence (PPIA). Statistical differences were tested by ANOVA.

Main results and the role of chance
Relative mtDNA abundance did not differ significantly (ANOVA p &gt; 0.05) between the three groups exhibiting different developmental potential (1±0.06 vs. 1.19±0.05 vs. 1.11±0.05, for Bl+Cl+ vs. Bl-Cl+ vs. Bl-Cl-, mean±s.e.m.).

Limitations, reasons for caution
Experiments were conducted in the bovine model. Although bovine folliculogenesis, monoovulatory ovulation and early embryo development exhibit considerable similarities with that of humans, caution should be taken when extrapolating these data to humans.
Wider implications of the findings: The use of molecular markers for oocyte developmental potential in cumulus cells could be used to enhance success rates following single-embryo transfer. Unfortunately, mtDNA in cumulus cells was not found to be a good proxy for oocyte quality.

Trial registration number
Not applicable
P–272 The aneuploid embryo secretome
Authors: Piccolomini M , Garcia C , Turco E L , Massaia I , Orteiro M , Duarte O , Yamakami L , Miyadahira E , Prado F 
Published: August 2021

Abstract

Study question
Does the metabolomic analysis of the embryonic culture medium predict the embryo aneuploidy?

Summary answer
The presence and quantity of some metabolites in the culture medium can select euploid embryos for transfer.

What is known already
Advances in analytical techniques for metabolomics have brought the possibility of better tools for the characterization of molecules. Embryonic metabolism can be used as a good indicator of viability, regardless of the morphology of the blastocysts, since differences were observed in the metabolic activities between the days of embryo development and in the rates of live births.

Study design, size, duration
17 patients had their embryos biopsied between January to July 2019 in a human reproduction laboratory. All cases had PGT-A indication and after the biopsy, the embryos were frozen. The culture medium samples were individually prepared for metabolites extraction according to the Bligh and Dyer protocol. Controlled ovarian stimulation and dose adjustments according to the response of each patient. The metabolomics analysis was performed by mass spectrometry.

Participants/materials, setting, methods
Ovum pick up will be performed 35 hours after r-hCG administration. The embryos were kept in individual 50ul drops until the blastocyst stage. The biopsy was performed in 26 blastocysts. The samples were sent to the 337 metabolites analysis by mass spectrometry. 15 molecules with the highest score on the PLS-Da was submitted the ROC curves to illustrate the power of the metabolic ploidy analysis. Besides, we performed the functional enrichment analysis for each group.

Main results and the role of chance
After the genetic analysis by PGT-a, 10 aneuploid embryos and 16 euploid embryos were found. Comparing the quantitative target metabolomic analysis of the 337 metabolites in the embryo culture medium, we observed the L-Alanine, Cytosine, Guanosine monophosphate, Homocysteine, Hypoxanthine, and Xanthine hiperrepresented in the aneuploid embryos, and the Citrulline, L-Glutamic acid, Kynurenine, L-Leucine, Methionine, Ornithine, L-Phenylalanine, L-Tyrosine, L-Valine were hiperrepresented in the euploid embryos. Through the ROC curve, we can verify AUC = 0.987. This result suggests that the analysis of euploid embryos through the metabolomic analysis of the culture medium is valid to be used as a noninvasive aneuploid diagnostic. The functional enrichment analysis shows the urea cycle and the glycine and serine metabolism as the principal function alter by aneuploid.

Limitations, reasons for caution
Small number of samples and not validate sample group.
Wider implications of the findings: Further studies are needed to validate these findings for the diagnostic of embryo euploidy.

Trial registration number
N/A