bluuebunny/crossref_metadata_2025 huggingface dataset:
P–109 Comparison between the outcome of sperm vitrification protocol and conventional slow freezing protocol for semen cryopreservation
Authors: Patel K , Sharma N , Mishra V , Aggarwal R , Suthar A , Sheth H
Published: August 2021

Study question

Does sperm vitrification technique helps in increasing sperm survival and low DNA fragmentation index post warming.

Summary answer

Sperm vitrification protocol results in better motility, high progression and low DNA fragmentation index as compared to slow freezing.

What is known already

Cryopreservation is ceasing and resuming the cell metabolism, which can be achieved by different techniques like slow freezing and vitrification .Vitrification allows solidification of the cells and extracellular milieu into a glass like state without formation of ice which protects intracellular and extracellular ice formation, and further helps in avoiding different types of cryo-injuries and cellular damage. Study design, size, duration: Comparative study from July 2019 to Oct 2020 in IVF unit of IKDRC Hospital. Two hundred and ten patients were randomized by computer generated list and divided into two groups. Group 1 (n = 110) samples cryopreserved by vitrification and Group 2 (n = 100) samples cryopreserved by conventional slow freezing.

Participants/materials, setting, methods

Semen sample were analyzed by WHO 2010 laboratory manual, including all normozoospermic samples , other abnormal samples were excluded from the study . Method of semen preparation before cryopreservation is similar for both the groups, double density gradient method of preparation was used . Semen sample with high viscosity, hypo and hyper-spermia were also excluded. Similar cryovials of 2ml volume were used for both groups.

Main results and the role of chance

In group 1 where samples were cryopreserved by vitrification sperm motility was (54.3% vs 49.2%)vs in group 2 where samples were cryopreserved by slow freezing , non- significant difference were observed , but progressive motility was significantly higher in group 1 as compared to group 2 (36.8%vs17.9%) and DNA fragmentation index is significantly lower in group 1 vitrification than in group 2slow freezing ( 9.7% vs 20%).

Limitations, reasons for caution

Technical proficiency of the operator to avoid human errors and still larger randomized control studies are needed to strengthen these results

Wider implications of the findings: Our study demonstrates that vitrification is better than slow freezing of human sperm, improved survival rates with high progression were found with vitrification and low DNA fragmentation index were also observed in samples cryopreserved with vitrification protocol.

Trial registration number

Not applicable
P–150 Does trophectoderm biopsy performed on different blastocyst stages affect the clinical outcome?
Authors: Tan S H , Chan A Q Y , Lim A Y X , Lim M W
Published: August 2021

Study question

The objective of this study is to evaluate the effect of trophectoderm (TE) biopsy on different blastocyst stages and its clinical outcome.

Summary answer

Our results showed that TE biopsy significantly reduced the clinical outcome of fully hatched blastocyst. What is known already: TE biopsy is a method widely practiced to harvest cells to determine the chromosomal constitution of a blastocyst, ensuring higher implantation and healthy pregnancies. The effect on clinical outcome after transferring blastocysts biopsied at different blastocysts stages has not been extensively studied.

Study design, size, duration

This retrospective study was conducted from January 2017 until July 2019 at Alpha IVF & Women’s Specialists. Following laser assisted hatching on day 3, TE biopsy was performed on unhatched, hatching and fully hatched day–5 blastocysts. A total of 1,020 single euploid blastocysts transfer (SBT) were performed. The average maternal age was 31.7. Implantation rates (IR) were evaluated for all stages of hatching (Unhatched: BG3 & 4; hatching: BG5; fully hatched: BG6).

Participants/materials, setting, methods

Laser assisted hatching (Hamilton Thorne Bioscience, USA) was performed on day–3 and subsequently cultured to blastocyst-stage. Different hatching stages were observed using embryoscope time-lapse system (Vitrolife, Sweden) and were recorded. Day–5 blastocysts with at least BG3BB grade (Gardner’s System) were selected for TE biopsy and the biopsied cells were sent for preimplantation genetic testing for aneuploidy (PGT-A) using Next-Generation Sequencing (Life Technologies, USA). All blastocysts were vitrified and warmed using the Cryotec Method (Cryotech, Japan). Main results and the role of chance: All 1,020 blastocysts survived post-warmed (post-warm survival rate= 100%) and were transferred in frozen transfer cycles. TE biopsy performed on unhatched blastocysts showed a comparable IR to hatching blastocysts (60.0% [15/25] and 65.2% [627/961]). While fully hatched blastocysts (44.12% [15/34]) show a significantly lower IR when compared to hatching blastocysts (65.2% [627/961]), no significant difference was seen when comparing unhatched blastocysts to fully hatched blastocysts (60.0% [15/25] and 44.12% [15/34]; p = 0.2949).

Limitations, reasons for caution

The sample size was comparatively smaller in the unhatched and fully hatched group than the hatching group. Further studies with a larger sample size is recommended to ascertain the clinical outcome. Since this is a retrospective study and biopsy was done by different embryologists, the biopsy technique was not controlled. Wider implications of the findings: To achieve higher clinical pregnancy, it is recommended to perform TE biopsy before the blastocysts is fully hatched.

Trial registration number

Not applicable
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

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 >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

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

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 < 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 < 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 < 0.001) and the inner cell mass (ICM, C1=35.0±2.4, C2=35.7±3.5, NEFA=26.0±2.2, P < 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 < 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

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

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

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

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

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
P–285 Controlling semi-invasive activity of human endometriotic stromal cells by inhibiting NF-kB signaling pathway using aloe-emodin or aspirin
Authors: Nasiri N , Babaei S , Moini A , Eftekhari-Yazdi P
Published: August 2021

Study question

Does inhibiting nuclear factor-kB (NF-kB) signaling by aloe-emodin (AE) or aspirin (Asp), as anti-inflammatory compounds, suppress the invasive activity of stage IV human endometriotic stromal cells?

Summary answer

Eutopic endometriotic stromal cells (EuESCs) seem to have a semi-invasive activity which is largely suppressed by AE or Asp.

What is known already

Inflammation and its master regulator, NF-kB, have been implicated in the development of endometriosis. Inhibition of NF-kB pathway using small molecules ameliorated disease progression and reduced the lesion size; nevertheless, underlying mechanism is not fully understood.

Study design, size, duration

In this cross-sectional study, a total of 8 infertile patients with proven endometriosis and 8 women without endometriosis (Control group) undergoing infertility treatment cycles, were enrolled between October 2018 and December 2019. The invasiveness of collected endometriotic stromal cells before and after treatment with AE or Asp, was analyzed and compared with the control group.

Participants/materials, setting, methods

The eutopic endometriotic and healthy endometrial biopsies were digested and the single cells were cultured. Gene and protein expression of proliferation, adhesion, and invasion markers of eutopic endometriotic stromal cells (EuESCs) with and without treatment with AE or Asp, as well as control endometrial stromal cells (CESCs) were analyzed using q-PCR and immunofluorescence staining, respectively. Cell migration capacity was assessed by wound closure assay.

Main results and the role of chance

We observed an association between NF-kB overexpression and higher proliferation/adhesion capacity in EuESCs. TNF-α, as a known NF-kB inducer, further potentiated this association. EuESCs at stage IV, displayed silent invasive and migratory behaviors. Pretreatment of EuESCs with AE or Asp significantly attenuated NF-kB expression and reduced proliferative, adhesive, invasive and migratory activity.

Limitations, reasons for caution

Due to some adverse effects observed following treatment with AE or Asp on the normal activity of EuESCs, more investigations on possible toxicity of the treatment, must be considered.

Wider implications of the findings: We suggest that both Asp and AE (as potent NF-kB inhibitors) may be useful as a supplement to conventional endometriosis treatments.

Trial registration number

Not applicable
P–350 Utilizing indocyanine green (ICG)-enhanced fluorescence to localize the ureter during robotic surgery for DIE and a concomitant crossed renal ectopia
Authors: Joukhadar R , Woeckel A , Altides A , Balafoutas D
Published: August 2021

Study question

Feasibility of (ICG)-enhanced fluorescence in visualizing the atypical course of the ureter during surgery for (DIE) of pelvic sidewall with a concomitant crossed renal ectopia.

Summary answer

Near-infrared fluorescence after transurethral injection of ICG enables localization of the ureter during surgery, thus facilitating complete excision of the lesions while enhancing patient’s safety.

What is known already

Existing case series refer to the transurethral injection of ICG and visualization under near-infrared (NIR) light during robotic surgery for real-time delineation of the ureter, which helps to prevent iatrogenic ureteral injury during complex surgery. The ICG reversibly stains the inside lining of the ureter by binding to proteins on urothelial layer. The consequent green fluorescence allows its identification throughout the entire case.

The presented case of a DIE of pelvic side wall along with an ipsilateral concomitant crossed renal ectopia (residual function 27%) resembles an utmost challenge for surgery. To our knowledge no similar case has been reported in literature.

Study design, size, duration

Demonstration of the Robotic technique by means of a step-by-step tutorial

Participants/materials, setting, methods

29-year-old patient referred after preceding laparoscopic surgery for DIE of left pelvic sidewall and abortion of surgery due to lack of accessibility/ severeness of the case. We performed renal scintigraphy, pelvic MRI and urological consultation.

Surgery was performed using an XI-da-Vinci robotic system. After cystoscopic placement of mono-Js we injected 4 ml. of ICG-solution (2,5 mg/ml). Visualization of the pelvic kidney was achieved 4 minutes after injection and of the complete ureter after 7–8 Minutes.

Main results and the role of chance

The robotic surgery could be completed safely and achieve a complete resection of the DIE of the pelvic sidewall including adhesiolysis of a broadly adherent bowel, opening of the rectovaginal space, ureterolysis of the distal portion of the ureter, partial excision of the left sacrouterine ligament and deperitonealization of the pelvic sidewall.

Postoperative controle revealed normal renal function and an adequate postoperative course.

Limitations, reasons for caution

ICG cannot be used in patients with iodine allergy.

Wider implications of the findings: Our report underlines the possibility to utilize indocyanine green (ICG)-enhanced fluorescence to localize the ureter during complex surgery for DIE, even in cases with atypical anatomy of the lower urinary tract.

Trial registration number

Not applicable
P–378 Using a machine learning tool (72% accuracy with 64% PPV) to predict multiple live birth when transferring multiple embryos, based on embryo specific data
Authors: Pena C A , Chambost J , Hickman C , Jacques C , Wiemer K , Kelley K
Published: August 2021

Study question

Can Machine Learning predict multiple pregnancy based on data specific to the embryos and the patient?

Summary answer

Embryo data are useful in determining which embryos are likely to lead to multiple pregnancy. Patient age has low predictive value compared to embryo data.

What is known already

Our previous assessment of the HFEA data demonstrated that single embryo transfer (SET) in the UK occurred in a minority (45%) of fresh cycles, with a marginal increase in live birth rate (LBR) in some patient cohorts in favor of multiple embryo transfer (MET). Current policies on determining number of embryos for transfer tend to be generic and do not account for detailed embryology data. Generic policies may compromise LBR for some patients that would benefit from MET. Artificial Intelligence has the potential to assist in this decision process.

Study design, size, duration

Retrospective cohort analysis from 2013 to 2020 of 193 cycles with 386 embryos used in double ETs on day 5 at POMA fertility clinic with positive live birth outcome. ML model, xgboost, was trained to predict multiple live birth (N = 54) versus single live birth (N = 139). Detailed embryology data from day 1 to day 5 were used as input.

Participants/materials, setting, methods

Input of the machine learning model included patient age and 18 morphological parameters collected on days 1, 2, 3 and 5 (symmetry, number of cells, blastocyst status, fragmentation, ICM and troph grades) from the two transferred embryos. An xgboost algorithm was trained on 80% of the data (n = 154) and tested on 20% of blind data (n = 39).

Main results and the role of chance

Xgboost machine learning algorithm predicted multiple live birth on the blind dataset with an accuracy of 72%, with an AUC of 0.60, showing better results than random. PPV (true prediction of multiple births) was 64% and NPV (true prediction of single birth) was 75%.

The following parameters ranked high in the predictive power of the machine learning (in order of predictive power): blastocyst status on day 5 of both embryos, symmetry on day 3, number of cells on day 2, scores on day 2 and 3. Limitations, reasons for caution: The dataset was derived from a single clinic with manual annotations and may not be transferable to other clinics. The risk of bias is important as the model was trained only àon embryos that were transferred and led to at least one birth

Wider implications of the findings: A tool to help identify which patients are at increased risk of MP with MET would be clinically useful to help patients and clinical team make the best personalised decision for a specific embryo, finding the balance between maximising success rate whilst minimising multiple pregnancy rate and its associated risks.

Trial registration number

Not applicable
P–390 The intrauterus administration of peripheral blood mononuclear cells increase the pregnancy rates for the patients with advanced maternal age after single euploid embryo transfer
Authors: Zhylkova E , Feskov O , Feskov V , Somova O , Zin Y , Yegunkova O
Published: August 2021

Study question

Does the intrauterine administration of peripheral blood mononuclear cells (PBMCs) effect the outcome of IVF for patients with advanced maternal age when the euploid embryos after PGT-A are transferred?

Summary answer

The implantation rates were significantly higher after the intrauterine application of PBMCs in patients with advanced maternal age (AMA) before transfer of the euploid embryo.

What is known already

The aneuploidy rates of blastocysts in IVF is in range 45–70% depending on different factors. Besides that, the endometrium plays an important role in achieving optimal outcomes of assisted reproductive technologies. It has been proposed that intrauterine administration of peripheral blood mononuclear cells modulates maternal immune response to favor implantation.

Study design, size, duration

The effect of the intrauterine application of PBMCs to improve the implantation rates in the group of patients with advanced maternal age was studied. Two group of patients (PBMCs-group and non-PBMCs-group_ were formed. Single euploid embryo was transferred for each patient. Participants/materials, setting, methods: The ploidy status of 373 blastocysts from 82 AMA-patients was analyzed by the method of next generation sequencing (NGS). PBMCs were applied for 39 women with the mean age 39.2±3.2 y.o. before embryo transfer (Group 1). For 43 patients with the mean age 38.2±2.1 y.o. single euploid embryo transfers were performed without PBMCs administration (Group 2). Chi-squared test was used for data analysis. The study’s protocol was approved by the Center’s IRB.

Main results and the role of chance

Totally the rate of euploid embryos was 27.1% (101 blastocysts). In the mentioned study 55.0% of examined blastocysts were aneuploid (205 embryos) and 17.9% of blastocysts were detected as mosaic (67 embryos). Single euploid embryo was transferred in each case in the patients of both experimental groups. The implantation rate was significantly higher in Group 1 with PBMCs application comparing with non-PBMCs experimental Group 2 (38.5% (15 pregnancies) vs. 23.3% (10 pregnancies), df = 1, χ2 = 5.487, χ2critical = 3.841, P = 0.020).

Limitations, reasons for caution

The embryo biopsy was performed only for blastocyst with top-quality morphology.

Wider implications of the findings: The implantation rates were significantly higher when the intrauterine application of PBMCs in patients with advanced maternal age before the transfer of the euploid embryo (P = 0.020). The randomized studies to improve our knowledge in immunogenic therapy in reproductive medicine should be performed.

Trial registration number

-
P–404 Low serum progesterone on the day of frozen blastocyst transfer is associated with a diminished ongoing pregnancy rate in hormonal replacement therapy cycles
Authors: Maignien C , Mathilde B , Valérie B , Ahmed C , Charles C , Pietro S
Published: August 2021

Study question

Is there a relationship between progesterone levels on the day of frozen blastocyst transfer and ongoing pregnancy rate (OPR), in hormonal replacement therapy (HRT) cycles?

Summary answer

Women undergoing HRT-frozen embryo transfer with progesterone levels≤9.76ng/ml on the day of blastocyst transfer had a significantly lower OPR than those with progesterone levels>9.76 ng/ml.

What is known already

The importance of serum progesterone levels around the time of frozen embryo transfer (FET) is a burning issue, in view of the growing number of FET worldwide. However, the optimal range of serum progesterone levels is not clearly determined and discrepancies arise from the current literature. Study design, size, duration: Observational cohort study with 915 patients undergoing HRT-FET at a tertiary care university hospital, between January 2019 and March 2020.

Participants/materials, setting, methods

Patients undergoing single autologous blastocyst FET under HRT using exogenous estradiol and vaginal micronized progesterone for endometrial preparation. Women were only included once during the study period. The serum progesterone level was measured in the morning of the FET, in a single laboratory. The primary endpoint was OPR beyond pregnancy week 12. Statistical analysis was conducted using univariate and multivariate logistic regression models.

Main results and the role of chance

Mean serum progesterone level on the day of FET was 12.90 ± 4.89 ng/ml). The OPR was 35.5% (325/915) in the overall population. Patients with a progesterone level ≤ 25th percentile (≤9.76ng/ml) had a significantly lower OPR and a higher miscarriage rate (MR) compared with women with progesterone level over Centile 25 (29.6% versus 37.4%; p = 0.033 and 34.8% versus 21.3%; p = 0.008, respectively). After adjustment for the potential confounders in a multivariate analysis, a serum progesterone level ≤ 9.76 ng/ml on the day of FETand FET of a Day 6-blastocyst (versus Day 5-blastocyst) were found as independent risks factor of lower OPR.

Limitations, reasons for caution

The main limitation of our study is linked to its observational design. Extrapolation of our results to other laboratories, or other routes and/or doses of administering progesterone also needs to be validated.

Wider implications of the findings: This study suggests that a minimum serum progesterone level is needed to optimize reproductive outcomes in autologous blastocyst FET, in HRT-cycles. Further studies are needed to evaluate if modifications of progesterone routes and/or doses may improve pregnancy chances, in an approach to individualize the management of ART patients.

Trial registration number

NA
P–448 Clinical outcome of social oocyte cryopreservation at advanced age
Authors: Tsafrir A , Ben-Ami I , Eldar-Geva T , Gal M , Weintraub A , Goldberg D , Dekel N , Levi H , Schonbeger O , Srebnik N , Nabulsi R , Buhbut I , Hyman J
Published: August 2021

Study question

What are the success rates of social oocyte cryopreservation (SOC) at advanced age?

Summary answer

In this study, one in four women who underwent SOC above age 35 had a delivery.

What is known already

While SOC is gaining popularity, reports on delivery rates are limited due to low utilization rates.

Study design, size, duration

Retrospective data collection of all woman who underwent SOC between 2011–2018, and presented for treatment using cryopreserved oocytes until January 2021. Participants/materials, setting, methods: Review of patient records (including both IVF and antenatal/postnatal) and laboratory data in a university affiliated hospital-based IVF unit. Main results and the role of chance: A total of 448 women underwent SOC during 2011–2018. 50 (11.2%) women returned to use these oocytes until the end of January 2021. Women who returned to use their oocytes underwent cryopreservation at mean age of 38.2±2.2. 46 (92%) of participants were above 35 at time of cryopreservation. Number of oocytes cryopreserved was 11.3±9.7. Mean time from cryopreservation to thawing was 5.5±1.8years (range 1–9 years). and age at thawing was 43.4±2.1 (range 40–49). Nearly half of patients initially attempted to conceive before using their cryopreserved oocytes, mostly by ART using fresh oocytes. Mean number of oocytes thawed and oocytes survived per women was 9.7±6.2 and 6.1±4.9 respectively (post thawing survival rate 65.4±35%).

Mean number of embryos transferred, at one or more attempts was 2.6±2.1 per women. Eleven women gave birth or had an ongoing pregnancy > 20 weeks at time of analysis. All deliveries resulted from cryopreservation at age 36 and older (delivery rate 23.9% per women). Limitations, reasons for caution: We report our initial experience of women who underwent SOC at a single center. Most women who returned to use their oocytes had undergone SOC at advanced age, therefore not necessarily reflecting outcome for younger patients attempting to preserve fertility using this technology. Wider implications of the findings: Considering modest success rates of SOC in our cohort, women considering SOC are advised to do so at an earlier age.

Trial registration number

Not applicable
P–518 Assessment of mitochondrial DNA viability ratio in day–4 biopsied embryos as an add-in to select euploid embryos for single embryo transfer
Authors: Metwalley A M , Hellani A , Esteves S , El-Damen A , Abde. Razik A , Dawood A A , Hamshary M E , Khamiss O
Published: August 2021

Study question

Is mitochondrial DNA viability ratio of day–4 biopsied embryos associated with embryo implantation potential?

Summary answer

The mitochondrial DNA viability ratio is significantly higher in embryos that implant. The score might help to select euploid embryos for single embryo transfer.

What is known already

Embryo euploidy is a critical factor for successful pregnancy outcomes. However, transfer of euploid embryos does not invariably result in implantation, thus indicating that other factors may play a role. Metabolic rates and adenosine triphosphate content vary significantly in oocytes and embryos and might affect embryo viability. Embryo function, indirectly measured by mitochondrial DNA viability ratio (mtV) has emerged as a potential quantitative biomarker for embryonic selection before the transfer, but clinical data remains limited. The purpose of this study is to characterize and compare mtV in euploidy day 4 embryos.

Study design, size, duration

Retrospective cohort study carried out between Jan. 2017 to Jan. 2020, involving 75 infertile couples undergoing IVF-ICSI with PGT-A and single embryo transfer (SET) of day 4 euploid embryos.

Participants/materials, setting, methods

We compared the mtV ratios of 34 non-pregnant patients with those of 41 patients who achieved clinical pregnancy after SET. The mtV ratio was obtained from a cohort of 75 euploidy embryos. The embryos were biopsied 80–85 hours post–ICSI and subjected to next-generation sequencing (NGS). The mtV was determined using Multiple of Mean (MoM) values, obtained by dividing the mtV ratio of individual embryos by the mean mtV value of all implanted embryos.

Main results and the role of chance

The mean mtV ratio (1.51; 95% confidence interval [CI] 1.25–1.77) of non-pregnant patients was significantly lower than those of pregnancy counterparts (2.5; 95% CI 1.82–2.68; p < 0.01). At a 0.5 MoM cutoff, the sensitivity and specificity of mtV ratio to discriminate between implanted embryos versus non-implanted embryos were 35.3% and 78.2%, respectively., with a positive predictive value (PPV) of 41.4%.

Limitations, reasons for caution

Our study is limited by the small sample size and lack of stratification by causes of female/male infertility. Endometrial receptivity issues, which could have contributed to implantation failure, was not evaluated.

Wider implications of the findings: Assessment of mtV ratio could provide additional prognostic information for selecting euploid embryos for transfer in SET programs. Further research is warranted to establish the clinical utility of routine application of mtV evaluation in PGT programs.

Trial registration number

N/A