bluuebunny/crossref_metadata_2025_split huggingface dataset:
P–155 Oocyte recovery 39 hours (from 39h to 41h) after administration of follicular maturation trigger does not affect clinical results
Authors: Michitaka K , Kitasaka H , Fukunaga N , Asada Y 
Published: August 2021

Abstract

Study question
What is the clinical outcome of oocytes recovered after 39 hours from ovulation inducing drug administration?

Summary answer
Oocytes obtained after 39 hours from follicular maturation triggering are equally viable to those obtained at the standard time of 36 hrs.

What is known already
In the clinical setting of ART, ovum pick-up (OPU) is generally performed around 36 hours after the administration of ovulation inducing drugs (OID). However, there are cases where OPU cannot be performed at this time often due to long operating lists. As the time elapsed between the administration of ovulation inducing drugs and OPU becomes longer, there is a concern about time-related oocyte aging. Nevertheless, there are few reports of clinical results of OPU after 36 hours from OID.

Study design, size, duration
We conducted a review of 1187 cycles and 1951 patients in which OPU and embryo transfer was performed in 2017–2018. All cycles underwent a ‘freeze-all’ of embryos and the transfer cycle was in the thawed embryo transfer cycle for all cases.

Participants/materials, setting, methods
The time from the administration of OID to the end of OPU was divided into 36h group and over 39h group and the MII and normal fertilization rate of oocytes obtained from OPU after ovarian stimulation were compared. After confirmation of fertilization, the D3 good-quality embryo and the D5 and 6 good-quality blastocyst rates of embryos that continued to be cultured and the pregnancy and miscarriage rates of cleavage-stage embryos and blastocyst transfers were compared.

Main results and the role of chance
The MII rate in the 36h and &gt;39h groups was 78.1% vs. 80.0%, and the normal fertilization rate was 77.9% vs. 78.1% (ICSI) and 65.4% vs. 67.6% (Conventional-IVF). The D3 good-quality embryo rate (good-quality embryos are embryos with less than 5% fragmentation in 7–9 cells and compaction with more than 50% adhesion between split spheres) was 21.8% vs. 25.3%, the D5 good-quality blastocyst rate (at least 3BB according to Gardner classification) was 33.6% vs. 40.1%, and the D6 good-quality blastocyst rate was 31.1% vs. 37.5%, all of which were not significantly different. The pregnancy rate for cleavage-stage embryo transfer was 26.6% vs. 6.7%, and the miscarriage rate was 25.3% vs. 42.9%, both of which were not significantly different. The pregnancy rate for blastocyst transfer was 45.4% vs. 50.0%, and the miscarriage rate was 22.2% vs. 20.0%, both of which were not significantly different. (The significance difference test was a χ-square test)

Limitations, reasons for caution
The study was a retrospective study.
Wider implications of the findings: Even if OPU is conducted after 36h of the administration of OID, to the extreme range of 39h–41h, oocyte aging does not seem apparent and pregnancy outcomes are similar to the standard time interval of 36 hours.

Trial registration number
‘not applicable’
P–164 Mulitcentre derived time lapse algorithms developed using 6228 transferred embryos with known birth outcome incorporating novel morphological and morphokinetic markers
Authors: Smith R , Petersen B , Barrie A , Montgomery S , Duffy S , Best L , Thirlby-Moore S , Wachter A , Kellam L , Campbell A 
Published: August 2021

Abstract

Study question
Can incorporation of novel markers of morphology with known temporal events successfully rank embryos to enable prediction of propensity for live birth?

Summary answer
Incorporation of variables for trophectoderm and morula grading demonstrably enhanced the model to rank embryos in order of potential for live birth.

What is known already
Models built using morphokinetic markers of development are widely used to rank embryos within a cohort. Such models include defined temporal parameters which are closely related to morphological grade. However, morphological grading by an embryologist is subjective and is not strongly correlated to outcome. Combining with defined kinetic events has been suggested to improve prediction of outcome.

Study design, size, duration
Data from 6228 known live birth outcome embryos from 8 UK clinics between 2011 – 2018 were investigated using an exploratory approach to identify novel markers of development.

Participants/materials, setting, methods
Five significant variables were defined, a derivative of time to start of blastulation; a derivative of trophectoderm grade; a kinetic variable utilising t3, t4, t5 and t8; an interval variable of tB-tSB and a variable based on novel morula classification. To maximise the output, a proxy value was derived for missing datapoints. The model was built using logistical regression and validated using fivefold cross validation with the data split as 80% training and 20% test.

Main results and the role of chance
An algorithm was developed including the five significant variables identified with an AUC of 0.685 demonstrating reliable prediction of live birth. Without morphological variables, the AUC was 0.674 demonstrating the improvement in the prediction value by including the derivative of the trophectoderm and morula grade. This resulted in ten classes of algorithm scores, 1–10, giving a live birth rate from 2% to 46%, irrespective of patient variables, for chance of live birth.

Limitations, reasons for caution
Successful application of the algorithm is reliant on stringent quality assurance for maintenance of accurate annotation and grading, and may not be transferable between laboratories with different SOPs.
Wider implications of the findings: The addition of a trophectoderm and morula grade in combination with morphokinetic parameters, increases the predictive value of the algorithm in relation to live birth outcome. Using proxy values allows maximization of data for model generation, and allows the model to be applied when missing values are present.

Trial registration number
Not applicable
P–176 Effects of high non-esterified fatty acids exclusively during bovine in vitro fertilization on cell lineage allocation of blastocysts
Authors: Idriss A , Okello E , Roger S , Velazquez M 
Published: August 2021

Abstract

Study question
Are there effects of high non-esterified fatty acids exclusively during bovine in vitro fertilization on cell lineage allocation of blastocysts?

Summary answer
Under the conditions of the present study, high exposure to NEFA during bovine IVF significantly decreases embryo production and alters cell allocation of resultant blastocysts.

What is known already
Cattle models have shown that a high exposure to non-esterified fatty acids (NEFA) such as steric acid (SA), palmitic acid (PA]) and oleic acid (OA) during in vitro oocyte maturation and embryo development can disrupt both embryo formation and quality. However, the fertilization process per se have been less studied, which is needed to identify developmental stages were potential therapies could be develop to ameliorate NEFA toxicity during the periconceptional period

Study design, size, duration
Day–8 blastocysts were immunostained for CDX2, a transcription factor involved in trophectoderm differentiation, to examine cell allocation of blastocysts derived from oocytes fertilised under high NEFA levels. VF (19 h) was carried with different NEFA levels(4 replicates) representing physiological (Control–1[C1], 28μM SA, 23μM PA, 21μM OA) and pathophysiological (NEFA, 280μM SA, 230μM PA, 210μM OA) relevant concentrations. A second control (C2) group contained solvent. Blastocysts (C1; n = 14, C2; n = 12, NEFA; n = 8)

Participants/materials, setting, methods
All blastocysts were examined by confocal microscopy and cell counting was done with the Imaris software. Data were analysed by ANOVA (mean±SEM) with percentage data arcsine transformed before analysis.

Main results and the role of chance
Blastocyst formation was decreased by high NEFA levels (C1=25.6±2.7%, C2=26.0±2.3%, NEFA=9.4±0.4%, P &lt; 0.001) which was associated with a decreased cleavage rate (C1=70.1±6.5%, C2=71.5±3.1%, NEFA=42.5±4.1%, P = 0.006) and an increase in embryo degeneration (C1=47.6±3.5%, C2=47.7±5.8%, NEFA=63.0±4.9%, P = 0.05). A lower total cell (TC) the number was observed in high NEFA-derived blastocyst (C1=125.2±6.6, C2=132.3±8.4, NEFA=67.3±5.6, P &lt; 0.001) associated with a low cell number in both the trophectoderm (CDX2 positive cells, C1=90.2±5.9, C2=96.7±6.4, NEFA=41.3±4.1, P &lt; 0.001) and the inner cell mass (ICM, C1=35.0±2.4, C2=35.7±3.5, NEFA=26.0±2.2, P &lt; 0.001). Furthermore, high NEFA-derived blastocyst showed an increased allocation of cells towards the ICM (ICM/TC proportion, C1=28.2±1.7%, C2=26.9±1.8%, NEFA=39.1±2.2%, P &lt; 0.001)

Limitations, reasons for caution
It will be better if the number of blastocysts reached increases.
Wider implications of the findings: such research can be widely applied to the human model due to the similarities between both specie.

Trial registration number
Royal Embassy of Saudi Arabia Cultural Bureau
P–180 Bisphenols are present in culture media used for ART and cell culture
Authors: Vignault C , Togola A , Desmarchais A , Téteau O , Maillard V , Bristeau S , Binet A , Guérif F , Elis S 
Published: August 2021

Abstract

Study question
Do plastic laboratory consumables and cell culture media used in human ART contain bisphenols?

Summary answer
Human embryo development media contained bisphenols close to the nanomolar concentration range while no release of bisphenols by plastic consumables was detected under routine conditions.

What is known already
The deleterious effect of the endocrine disruptor bisphenol A (BPA) on female fertility raised concerns regarding ART outcome. BPA was detected neither in media nor in the majority of plastic consumables used in ART, however it might have already been replaced by its structural analogs, including bisphenol S (BPS).

Study design, size, duration
Seventeen plastic consumables and 18 cell culture and ART media were assessed for the presence of bisphenols.

Participants/materials, setting, methods
Ten different bisphenols (bisphenol A, S, AF, AP, B, C, E, F, P, and Z) were measured using an isotopic dilution according to an on-line solid phase extraction / liquid chromatography/mass spectrometry method.

Main results and the role of chance
While all the plastic consumables of this study did contain bisphenols, none of them did release bisphenols under routine conditions. Moreover, 16 of the 18 cell culture and ART media assessed contained bisphenols, including 8 among the 10 media used in human ART. Five human ART media exhibited bisphenol concentrations higher than 0.8 nM and reached up to 3.2 nM (799 ng/L).

Limitations, reasons for caution
Further studies are required to investigate a greater number of ART media to identify less potentially harmful ones, in terms of bisphenol content.
Wider implications of the findings: As BPS has already been reported to impair oocyte quality at nanomolar concentrations, its presence in ART media, at a similar concentration range, could contribute to a decrease in the ART success rate. Thus far, there has been no regulation of these compounds in the ART context.

Trial registration number
Not applicable
P–196 Bacterial influence on oocyte quality - the secret of a successful fertilization
Authors: Schenk M , Voroshilina E , Boldyreva M , Koranda M , Reinschissler N , Weiss G 
Published: August 2021

Abstract

Study question
Is there a difference in bacterial composition of follicular fluid between oocytes developing a good quality blastocyst and oocytes that fail fertilization?

Summary answer
Follicular fluids of oocytes failing fertilization show a different bacterial profile compared to follicular fluids of oocytes that were successfully fertilized.

What is known already
The presence of pathogens in the female reproductive tract has been intensively investigated. Lactobacillus species are mainly associated with a healthy genital tract and good prognosis for a successful pregnancy. Studies of the bacterial composition of follicular fluids have been mainly undertaken in women participating in reproductive medicine treatment because of the nature to obtain the specimen. In most studies follicular fluids have been pooled for analysis. Information on separately collected follicular fluids is still rare. We hypothesized that the composition of bacteria within follicular fluids is responsible for the success of the fertilization process.

Study design, size, duration
The study was designed and conducted at the Kinderwunsch Institut Schenk GmbH (Dobl, Austria) together with DNA-Technology. Follicular fluids from 46 patients undergoing IVF (in vitro fertilization) and ICSI (intracytoplasmic sperm injection) treatment were included and analyzed.

Participants/materials, setting, methods
Follicular fluids from 46 patients were collected separately. 2 follicular fluids from each patient were screened for common bacteria of the genital tract. One from an oocyte developing a good quality blastocyst and one displaying fertilization failures. Samples were analyzed for bacterial composition using the Femoflor16 (DNA-Technology).

Main results and the role of chance
Quantitative analysis revealed a higher total bacteria mass in follicles from oocytes that failed fertilization. Furthermore, Lactobacillus were not present in those follicles compared to good blastocyst follicles. In addition, Chlamydia trachomatis was found mainly in follicular fluid of not fertilized oocytes together with Eubacterium, Gardnarella and Trichomonas species. Interestingly, a trend of elevated levels of Ureaplasma species in follicular fluids of oocytes developing good quality blastocysts was observed.

Limitations, reasons for caution
Contamination of follicular fluids due to the procedure of oocyte pick up and follicular fluid retrieval cannot be completely excluded. Results should be confirmed with a higher sample size.
Wider implications of the findings: We assume that different bacterial compositions in follicular fluids are responsible for the destiny of the oocyte. It is tempting to speculate that bacterial analysis of follicular fluids may be beneficial to select to best oocytes in future IVF/ICSI treatments.

Trial registration number
Not applicable
P–205 Epothilone D as an actin cytoskeleton stabilizer improved mitochondria bioenergenesis and blastocyst formation of mouse preimplantation embryo
Authors: Cho M J , Kim Y J , Kim M J , Kim Y S , Park E , Choi K H , Kang J Y , Kim H O , Koong M K , Kim Y S , Yoon T K , Ko J J , Lee J H 
Published: August 2021

Abstract

Study question
What is primary factor of bioenergetics product activity between microtubule instability and the functional activity of mitochondria in embryo?

Summary answer
The actin cytoskeleton instability is presumably the primary cause for the bioenergenesis of mitochondrial function to the preimplantation embryo development.

What is known already
Mitochondria are cellular organelles dynamically moving and morphological changes. It provides for homeostatic energy to the cell. The dynamic property of the mitochondria is associated with the microtubule network in the cell. However, the stability of the microtubule was clearly identified for preimplantation embryo development.

Study design, size, duration
This study is designed to assess the ATP productivity of the mitochondria, and specifically to observe what its primary factor is in terms of providing microtubule stability in mammalian cells. Additionally, we investigated the relationship between blastocyst formation and actin cytoskeleton stabilization by EpD with 2-cell mice.

Participants/materials, setting, methods
We prepared the microtubule stability regulation model with the HEK293 cell line by using the microtubule stabilizer as an Epothilone D (EpD). Then we analyzed the metabolic activity of the cells through oxidative phosphorylation (OXP) ratios analysis. Also, we performed confocal live imaging to observe mitochondria morphology depending on the cells’ microtubule. Next, we treated EpD to 2-cell culture media for the analysis of blastocyst development ratios.

Main results and the role of chance
EpD significantly increased fusion form. Also, EpD enhance bioenergy ratios like OXP in the mitochondria and functional activity related marker, like mTOR compared with the control. These results suggest that microtubule stabilization enhances mitochondrial metabolism by increasing oxygen consumption. Also, EpD in 2-cell culture media led to a significant increase in the speed of development and 50% higher hatched out blastocyst formation ratios compared to the control group.

Limitations, reasons for caution
This study had limited animal experiments. For the next study, we are planning with an aim to improve the quality and development ratios of human embryos by EpD.
Wider implications of the findings: Microtubule stabilizer has a possibility to recover the mitochondria’s functional activity in the preimplantation embryo development. Mitochondrial functional activity along the actin cytoskeleton may play a pivotal role in determining the embryo quality and development ratios for archive pregnancy.

Trial registration number
non-clinical trials
P–222 Can we optimise the time that we perform the fertilisation check in the lab? Lessons learnt from time-lapse incubation
Authors: Barrie A , Smith R , Best L , Davis N , Duffy S , Krokos S , Lodge Y , Montgomery S , O’Boyle S , Thirlby-Moore S , Whitten B , Campbell A 
Published: August 2021

Abstract

Study question
Can time-lapse data be used to identify the optimum time to perform the fertilisation check for oocytes cultured in standard incubation?

Summary answer
The optimum time to perform fertilisation checks for oocytes cultured in standard incubation is 16.5hpi+/–0.5h.

What is known already
Time-lapse incubation allows the embryologist to retrospectively review collated images of oocytes and embryos to capture important embryological observations that may have otherwise been missed. This is a luxury not available to embryologists when oocytes or embryos are cultured in standard incubation. Traditionally, the optimum time to perform the fertilisation check is 17 hours post insemination (hpi) +/- 1 hour. It was hypothesised that this could be fine-tuned ensuring the maximum number of fertilised oocytes were observed, thereby increasing the number of usable embryos for the patient.

Study design, size, duration
This was a retrospective, multicentre analysis including data from 27,022 ICSI derived embryos cultured in time-lapse incubation between January 2011 to November 2019.

Participants/materials, setting, methods
The time of pronuclei appearance and disappearance was recorded using the time-lapse incubation software. The number of oocytes exhibiting normal fertilisation (defined as the presence of two pronuclei) during 30 minute intervals from 15hpi to 20hpi was determined.

Main results and the role of chance
Between 15–17.5hpi the average number of oocytes exhibiting normal fertilisation was 98.19% with most oocytes having visible pronuclei at 16–16.5hpi (98.32%). At 18–18.5hpi the number of visible pronuclei falls to 95.53% and continues to fall to 87.02% at 19.5–20hpi meaning that over 3000 (11%) normally fertilised oocytes, within this cohort, would not be identified.

Limitations, reasons for caution
The conclusions of this investigation cannot be effectively extrapolated to IVF embryos as only ICSI embryos were used for the determination of the results.
Wider implications of the findings: The optimum time to perform fertilisation checks for oocytes cultured in standard incubation is 16.5hpi+/–0.5h. However, without the use of time-lapse incubation, the fertilisation of at least 2% of embryos that create a fetal heart will be missed, even if the fertilisation check is performed in the optimum window (16.5hpi+/–0.5h).

Trial registration number
Not applicable
P–236 The evaluation of apoptosis and luteinization process in cumulus cell culture of IVF patients in terms of embryo development and pregnancy
Authors: Dünda. Çiftlik G , Ergüven M , İrez T 
Published: August 2021

Abstract

Study question
Is there any relationship between apoptosis rate and progesterone levels in terms of the degree of embryo quality and clinical pregnancy success?

Summary answer
The cumulus cell apoptosis and progesterone levels measured at the hCG day were associated with embryo quality and clinical pregnancy success.

What is known already
The high rate of apoptosis at cumulus cells led to poor embryo development with the clinical pregnancy failure.

Study design, size, duration
It is prospective study carried out with 40 healthy women diagnosed as the male factor between September 2017 and April 2018.

Participants/materials, setting, methods
40 healthy women aged between 27–38 and diagnosed as the male factor who were undergoing IVF participated the study. The cumulus cells were taken after denudation and cultured. The rate of the apoptosis were measured using with TUNEL method every 24 hours for 96 hours. In concominant with the apoptosis evaluation, the embryo progession and clinical pregnancy rate were also monitored. Progesterone levels taken at the hCG day were also measured.

Main results and the role of chance
In this study, it was shown that cumulus cell apoptosis was lower in the pregnant group compared to the non-pregnant group (p = 0.023), and progesterone value on the hCG day was higher in non-pregnant group (p = 0.021). From the data obtained at the end of the 4th day of culture showed that the apoptosis rate showed a higher tendency in non-pregnant women and this was statistically significant (p = 0.009). In our study, it was found that the ratio of apoptotic cells up to the 4th day showed a positive correlation with the progesterone levels on the hCG day (p = 0.001).
Briefly, these results were obtainedfrom this current study that 1) The increase in the rate of apoptotic cells of cumulus cell culture has a negative effect on pregnancy rates, 2) The progesterone levels measured on the day of hCG has a significant negative effect on pregnancy, 3) The increase in progesterone levels acts as a messenger about the apoptosis rate of cumulus cells, 4) The luteinization is strictly associated with the apoptosis of cumulus cells. Limitations, reasons for caution: This was a single center study. Results need to be validated across different centers and high numbered different study population.
Wider implications of the findings: The increase in progesterone value acts as a messenger about the rate of apoptosis of cumulus cells may be the prominent indicator to obtain high grade embryo with high clinical pregnancy success.

Trial registration number
2017/5–8