bluuebunny/crossref_metadata_2025 huggingface dataset:
O-055 Why we should be testing embryos for polygenic traits
Authors: Savulescu J
Published: August 2021

Abstract text

The past few years have brought significant breakthroughs in understanding human genetics. This knowledge has been used to develop ‘polygenic scores’ (or ‘polygenic risk scores’) which provide probabilistic information about the development of polygenic conditions such as diabetes or schizophrenia. They are already being used in reproduction to select for embryos at lower risk of developing disease. The world’s first baby, Aurea, has been born after being selected on the basis of a Genomic Health Index score provided by Genomic Prediction (https://www.youtube.com/watch?v=HE5ADe7BgdM&ab_channel= GenomicPrediction).

I argue for the use of this technology not only for the selection of embryos at lower risk of disease but for the selection of advantageous non-disease traits.

Currently, the use of polygenic scores for embryo selection is subject to existing regulations concerning embryo testing and selection. Existing regulatory approaches include ‘disease-based’ models which limit embryo selection to avoiding disease characteristics (employed in various formats in Australia, the UK, Italy, Switzerland and France, among others), and ‘laissez-faire’ or ‘libertarian’ models, under which embryo testing and selection remain unregulated (as in the USA). With Sarah Munday, I have introduced a novel ‘Welfarist Model’ which limits embryo selection according to the impact of the predicted trait on well-being. We compare the strengths and weaknesses of each model as a way of regulating polygenic scores. Polygenic scores create the potential for existing embryo selection technologies to be used to select for a wider range of predicted genetically influenced characteristics including continuous traits. Indeed, polygenic scores exist to predict future intelligence, and there have been suggestions that they will be used to make predictions within the normal range in the USA in embryo selection. We examine how these three models would apply to the prediction of non-disease traits such as intelligence. The genetics of intelligence remains controversial both scientifically and ethically. This paper does not attempt to resolve these issues. However, as with many biomedical advances, an effective regulatory regime must be in place as soon as the technology is available. If there is no regulation in place, then the market effectively decides ethical issues.
O-141 Mapping of SARS-CoV-2-associated receptors and proteases mRNA in human endometrium during natural and stimulated cycles
Authors: Haouzi D , Entezami F , Brouillet S , Barry F , Gala A , Ferrieres-Hoa A , Tal A , Hamamah S
Published: August 2021

Study question

Covid-19 pandemic has significantly affected the assisted reproductive technology (ART) practice. Understanding whether SARS-CoV-2 could infect endometrial tissues during ART is crucial for risk mitigation

Summary answer

Analyses of gene expression profiles of SARS-CoV-2 host entry candidates from microarray data suggest that endometrium should be considered as potential target for SARS-CoV-2 infection.

What is known already

Very few studies analyzed the gene expression profiles of SARS-CoV-2-associated receptors and proteases, mainly focusing on ACE2 and TMPRSS2 expression, resulting incomplete knowledge in different specimens from female genital tract. However, no studies have analyzed the potential impact of controlled ovarian stimulation (COS) protocols during ART procedure on the endometrial gene expression profiles of SARS-CoV-2-associated receptors and proteases

Study design, size, duration

To address this question, we retrospectively examined the gene expression profile of SARS-CoV-2-associated receptors and proteases in endometrial biopsies of a cohort of ART candidates using Affymetrix microarray data

Participants/materials, setting, methods

Human endometrial tissue under natural (n = 62) and COS cycles (n = 42) were analyzed. A focus was particularly made on the renin-angiotensin system relates genes with a prominent role in the virus infection, and gene expression levels of receptors and proteases closely related to SARS-CoV-2 infectionwas also studied.

Main results and the role of chance

Using our large cohort of endometrial samples, we reported a high prevalence of genes related to the ACE2 pathway, including AGT, AGTR1, ANPEP, CTSA, ENPEP, LNPEP, MME, NLN, THOP1, BSG and CTSL during both phases(early- and mid-secretory phase), and mainly during the mid-secretory phase for ACE2. The highest signal intensities were found for CTSA, LNPEP, MME, NLN, BSG and CTSL. The most representative of dual coexpression of SARS-CoV-2-associated receptor and protease in endometrium was BSG-CSTL and BSG-CTSA. It s also important to note high variation of SARS-CoV-2 receptors inter-patients under natural cycle.Globally, the impact of COS on endometrial gene expression profile of SARS-CoV-2-associated receptors and proteases of non Covid-19 patients is low, suggesting no additional potential risks of SARS-CoV-2 infection during stimulated ART procedure compared with natural cycles.

Limitations, reasons for caution

Analyses of Affymetrix microarray gene expression data were performed in non-COVID-19 patients. Whether the SARS-CoV-2 infection changes the endometrial gene expression profile of SARS-CoV-2-associated receptors and proteases is under investigation

Wider implications of the findings

Specimens from female genital tract may be considered as potential targets for SARS-CoV-2.

Trial registration number

not applicable
O-148 Sperm Aminopeptidase N as a predictive biomarker of blastocyst development and embryo viability
Authors: Subiran N , Urizar-Arenaza I , Muñoa-Hoyos I , Irazusta J , Larreategui Z , Garrido N , Gianzo M
Published: August 2021

Study question

To evaluate human sperm APN as a prognostic factor for determining high-quality embryos.

Summary answer

The human sperm APN has the potential to become new molecular prognostic biomarker for having high-quality and viable embryos.

What is known already

Prognosis and diagnosis of male fertility is one of the major concerns in reproductive medicine. Approximately 30%-40% of men with otherwise normal fertility parameters are still unable to achieve pregnancy. The predictive clinical value of a semen analysis to identify fertile or infertile males is limited; therefore, new sperm diagnostic or prognostic methodologies are urgently required. Sperm Aminopeptidase N (APN) may be a relevant molecular marker due to its high concentration in sperm cells and its role in sperm physiology, such as motility, acrosome reaction, and embryo development.

Study design, size, duration

A prospective study that involves a total of 81 couples and 611 embryos who underwent oocyte-donation cycles at the Clínica IVI Bilbao (Spain) between September 2014 and July 2015.

Participants/materials, setting, methods

This study was set in an assisted reproduction unit and in an academic research laboratory. All semen samples were examined and classified following WHO guidelines. Spermatozoa were isolated from semen on discontinuous colloidal silica gradient (45%-90%) technique. Embryo quality and development were determined according to the Spanish Association of Reproduction Biology Studies (ASEBIR) criteria. Flow cytometry analyses of quantitative and semi-quantitative sperm human APN levels.

Main results and the role of chance

The obtained results proved that the most evolved and viable blastocysts were associated with low sperm APN levels. Expanding, expanded, hatching/hatched and viable blastocysts come from semen samples which showed lower APN levels than early blastocysts, blocked and non viable blastocyst. The cumulative probability of having more evolved blastyocysts increased 1.38-fold at day 5 and 1.98-fold at day 6 of embryo development as well as the likelihood of having viable embryo increased 1.48-fold when semen samples with low APN levels are used during the ICSI technique.

Limitations, reasons for caution

Data obtained from a single Fertility Clinic. A multi-centrum study will be required.

Wider implications of the findings

The human sperm APN has the potential to become new molecular prognostic biomarker for having high-quality embryos that could help to diagnose male infertility, especially when seminal parameters are close to the threshold values.

Trial registration number

Not applicable
O-158 The effect of circadian rhythm disruption due to constant light on ovarian and oocyte aging through possible relationship between PER2 and mTOR signaling pathway
Authors: Bora G , Onel T , Yildirim E , Yaba A
Published: August 2021

Study question

Does circadian rhythm disruption by constant light affect the ovarian morphology and function, and cause ovarian and oocyte aging through possible relationship between PER2 and mTOR?

Summary answer

We demonstrated that circadian rhythm disruption by light may cause ovarian and oocyte aging.

What is known already

Circadian rhythm regulates multiple physiological processes and PER2 is one of the core circadian rhythm components. Changes in light conditions may cause circadian rhythm disruptions. Light exposure at night may cause attenuation in PER2 mRNA and protein levels. Circadian rhythm disruptions are thought to be associated with reproductive diseases. mTOR signaling pathway functions in folliculogenesis and oocyte maturation in ovary. Also, it is associated with ovarian and oocyte aging.

Study design, size, duration

A total of 32 female Balb/c mice which enter estrous cycle were used in the study. Mice were randomly assigned to one of two groups as 12:12h L:D and 12:12h L:L. During the experiment, 12:12h L:D (control group) was housed in a 12:12h light:dark cycle and 12:12h L:L (experiment group) was housed in a constant light conditions 12:12h light:light for 1 week.

Participants/materials, setting, methods

We housed 12:12h L:D group in standard lightening conditions and 12:12h L:L group in constant light for one week. We performed food intake and body weight change analysis. We evaluated ovarian morphology, follicle counting analysis. We evaluated ZP3 and nitrotyrosine (NTY) expression for oocyte aging markers. We performed western blot for PER2, mTOR, p-mTOR, p70 S6K, p-p70 S6K, and Caspase-3 protein levels.

Main results and the role of chance

We demonstrated that circadian rhythm disruption caused alteration in their food intake and decrease in primordial follicle numbers and increase in atretic follicles (p < 0.05). It caused increase in oxidative stress and decrease in ZP3 expression in oocytes (p < 0.05). We showed decreased protein levels of PER2, mTOR, p-mTOR and p70 S6K (p < 0.05). 

Limitations, reasons for caution

The explanation of molecular mechanism underlying the relationship between circadian rhythm disruptions by light and ovarian function may lead the usage of circadian rhythm-based or light-based therapies currently using to treat some diseases on female reproductive system related diseases.

Wider implications of the findings

We conclude that constant light may reduce follicle reserve, cause follicles to go rapidly atresia and disrupt the oocyte quality, thus it may be a risk factor for female reproductive diseases such as premature ovarian insufficiency and early menopause. 

Trial registration number

not applicable
O-058 The impact of endometriosis on fertility treatment
Authors: Somigliana E
Published: August 2021

Abstract text

Endometriosis can affect natural fertility by distorting pelvic anatomy and by causing local pelvic inflammation that affects the quality of the oocytes, the function of the tubes and the sperm capacity to fertilize. Two approaches can be foreseen, either removing the disease with the aim of restoring normality (surgery) or overcoming the disease by retrieving oocytes in the ovary and allowing their fertilization in in vitro conditions (IVF). Both therapeutic approaches seem rational and wise. However, despite this apparently simple clinical scenario, management of endometriosis-related infertility has engendered long-lasting and burning debates over the last two decades and, to date, no consensus has been reached. Robust scientific evidence is scant, actually limited to the demonstration that laparoscopy modestly increases the chance of natural conception.

In this debate, it is noteworthy that chances of pregnancy do not represent the unique stake. Other factors can play a role and deserves consideration. Considering the choice of fertility treatment, he most relevant are the following: 1) the impact of ovarian endometriomas and their removal on ovarian responsiveness, 2) the impact of endometriomas and endometriosis in general on oocytes quality, 3) the additional risks of oocytes retrieval in the presence of endometriosis (infections in particular) 4) the detrimental effects of endometriosis-related hydrosalpinx, 5) the impact of endometriosis on uterine motility and endometrial receptivity, 5) the confounding effect of endometriosis-associated adenomyosis. Unfortunately, for the vast majority of these concerns, evidence is not definite and physicians have to take decisions based on their clinical commonsense.
O-060 Ethical implications of the direct to consumer genetic testing. What should donors and recipients know?
Authors: Frith L
Published: August 2021

Abstract text

The growth in the use of direct-to-consumer-genetic testing (DTCGT) is having a major impact on sperm, egg and embryo donor conception (hereafter donor conception). DTCGT services include family history sites, e.g. Ancestry.com, and medical testing sites, e.g.23andme. Despite the many different motivations people have for using these services, it is now easier to search and find donor relatives, with donor-conceived people, recipients of donor gametes and embryos, and donors all using these services to make hitherto unlikely connections. Some individuals have found large numbers of donor-siblings, while donors have been traced by their adult donor offspring. DTCGT can also reveal unexpected origins with the numbers of people finding out they are donor-conceived through DTCGT rapidly increasing. For example, one woman discovered she was donor conceived after using 23andme to assess her risk of breast cancer, an eventuality she had never anticipated when she decided to take that test.

The increasing use of DTCGT has led to claims that donor anonymity is dead and we are entering a new era where the possibilities of finding our genetic relatives and extended family have dramatically expanded. These developments will produce new landscapes where different systems collide and interact, creating new ways of locating and finding donor relatives. In the UK for example, information on genetic relations, donors and donor siblings, is located within two very different systems: ‘official’ regulatory systems, such as central registers of information held by government bodies such as the Human Fertilisation and Embryology Authority’s (HFEA) registers; and emerging digital online systems, of DTCGT.

This paper will explore how these new developments interact with existing ways of finding out information about donor relatives and consider the ethical and legal issues and challenges for fertility practice.
O-176 Secondary ovarian insufficiency (SOI) - a new infertility diagnosis
Authors: Gleicher N , Darmon S K , Albertini D F , Barad D H
Published: August 2021

Study question

Can, in contrast to primary ovarian insufficiency (POI), ovarian insufficiency also be caused as a secondary event (SOI)?

Summary answer

Adrenal hypo-androgenism, caused by insufficiency in androgen production by the zona reticularis, may mimic POI in clinical presentation.

What is known already

A variety of small and large animal models have, especially at small growing follicle stages, conclusively demonstrated the dependence of normal follicle maturation on adequate testosterone levels, a process mediated via the androgen receptor on granulosa cells. Increasing clinical evidence, moreover, has demonstrated clinical improvements in oocyte numbers and oocyte quality following androgen supplementation at these stages in hypo-androgenic women with premature ovarian aging (POA). Whether adrenal hypo-androgenism in extreme cases can, however, also lead to complete ovarian insufficiency mimicking POI, is unknown.

Study design, size, duration

Based on FSH levels >40.0 mIU/mL and amenorrhea, we searched our center’s electronic research data bank for patients who between 2016-2018 presented with a diagnosis of presumed POI.

Participants/materials, setting, methods

Among 78 POIs, 13 demonstrated low DHEAS (< 100 ug/dL), i.e., adrenal hypo-androgenism, 6 rejected egg donation and received DHEA (Fertinatal®, 25mg TID, Fertility Nutraceuticals, LLC, New York, N.Y., USA) and Coq10 (Ovoenergen®, 333mg TID,same ­manufacturer) before stimulation with 450IU FSH and 150 IU hMG (different ­manufacturers). All women were < age 41, demonstrated menopausal FSH (>40.0 mIU/mL), undetectable AMH, amenorrhea, > prior cycle cancellation, elevated SHBG, low total testosterone and low DHEAS (high DHEA/DHEAS).

Main results and the role of chance

Out of the 6 patients so stimulated, 5 demonstrated follicular responses following DHEA supplementation and 2/6 conceived spontaneously and delivered healthy offspring. One of these two is currently in treatment for another child.

Limitations, reasons for caution

To better understand adrenal control of ovaries via androgen production, further elucidation of endocrine signaling between adrenals and ovaries is required, including detection of unknown, though increasingly likely, feedback loops, considering that adrenals and ovaries share a primordium.

Wider implications of the findings

We confirmed existence of SOI due to adrenal hypoandrogenism, defined its phenotype, demonstrating that some patients with alleged POI actually exhibit SOI. Better pregnancy chances with adrenal SOI than POI, reemphasize the importance of correct differential diagnoses to avoid premature referrals of women into third-party egg donations

Trial registration number

N/A
O-068 Training of clinicians: How to do it and how to best evaluate it
Authors: Feki A , Tatjana M , Roy F
Published: August 2021

Training of clinicians: How to do it and how to best evaluate it?

Anis Feki, Tatjana Motrenko, Roy Farquharson

Many countries within Europe and EU do not have a specific national agency that provides the necessary structure or governance for appropriate training in Reproductive Medicine (RM). Therefore, The European Society of Human Reproduction and Embryology (ESHRE) has been the responsible agency for accreditation of RM training alongside its sister organization, the European Board and College of Obstetrics and Gynaecology (EBCOG). Both of these organizations are ultimately responsible to and approved by the Union of European Medical Specialties (UEMS), which represents the European Union (EU) governing body for medical practitioners’ education.

The Reproductive Medicine subspecialist is a specialist in basic obstetrics and gynecology who has undergone theoretical and practical training in the medical and surgical management of infertility, including assisted reproductive techniques (ART). In general, the comprehensive management of these problems includes both diagnostic and therapeutic procedures allied to continuous audit of outcome.

The global aim is to improve the care of patients with disorders of reproductive function. To reach this target, both ESHRE and EBCOG put in place a program that starts with accreditation of training centers in obstetrics and gynecology and more specifically, specialist training centers in reproductive medicine. Fellows must have a structured program of a minimum of 2 years after completing their basic obstetrics and gynecology syllabus and/or obtaining EFOG European Fellow of Obstetrics and Gynecology) diploma for non-European basic training. When a training fellow in RM has completed their logbook, the fellow will finish by assessing an individual’s knowledge and skills by passing an exit-training exam called the EFRM (ESHRE-EBCOG Fellow in Reproductive Medicine) diploma.

The success of and the need for such certification has led ESHRE to structure the training also for reproductive surgeons, embryologists, as well as nurses and midwives. In addition, several countries within the EU already have a national compulsory specialist accreditation system for RM training.

Recent proposals by the EU Commission to build European reference networks across all specialties, including RM may require the recognition of ESHRE/EBCOG–accreditation as an appropriate entry point for consideration of diagnostic and other interventions. Encouraging subspecialists and centers to apply for training accreditation may well prove to be an increasingly attractive option as educational needs across Europe become integrated and formalized through UEMS. ESHRE holds the view that accredited centers/subspecialists for training are beacons of excellence that often go hand in hand with high-quality research.
O-199 Proof of concept: implantation window must be wider than proposed. Report of seven twins after asynchronous double embryo transfer
Authors: Vega A. Gosálvez , Mazaira M. Rodriguez , Fernandez N. Martin , Nuñez M. Iglesias , Brandt M , Juan L. Vidal , Vargas P. Pastor , Rodriguez Z. Gonzalez , Bermudez S. Corral , Seivane R. García-Abadillo , Abbad R. Sainz de la Cuesta
Published: August 2021

Study question

Can simultaneous transfer of two embryos that were cryopreserved at different stages (D3 and Blastocyst) be appropriate to enhance success in women with more than three failed embryo transfers?

Summary answer

Double asynchronous embryo transfer offered excellent results in RIF. Unexpectedly high twin rate suggests that embryo-endometrium synchrony is overemphasized. Implantation window must be wider.

What is known already

Transcriptomic signature of the endometrium has been investigated in the last few years trying to understand the best moment for embryo implantation. Nevertheless, the optimal period has not been well established yet in humans. Simultaneous transfer of two human embryos at different developmental stages (D3 and Blastocyst) on Day 4 was proposed to help couples who have had RIF.

Study design, size, duration

Observational case-control study. From April 2016 to January 2021, we offered double asynchronous embryo transfer only after Recurrent Implantation Failure (RIF). Two requirements were necessary: 1) Double embryo transfer was acceptable by the couple due to poor reproductive outcome. 2) Availability of two embryos cryopreserved at different stage (D3 and Blastocyst). Results were compared with good prognosis patients (all patients under 35 years in that period who had elected to transfer two day 3 cryopreserved embryos).

Participants/materials, setting, methods

Forty-five patients accepted to participate in the study. Results were compared with all patients (237) under 35 years where two D3 thawed embryos were transferred. All cases received same protocol (oral estradiol 6mg/d or vaginal estradiol 4mg/d until ultrasound showed endometrial growth) LH, P4 and E2 were monitored in all patients to detect spontaneous LH surge. All cases received transvaginal micronized progesterone 800 mg/d. Embryo transfers were ultrasound guided and Wallace Embryosure catheter was employed.

Main results and the role of chance

Limitations, reasons for caution

Multiple pregnancy rate was unacceptably high. Therefore, it should not be suggested for good prognosis couples where single embryo transfer is clearly advidsed. Our main limitation was the combination of D3 embryos with blastocysts. The retrospective design make the results to be considered as a proof of concept.

Wider implications of the findings

Double asynchronic embryo transfer can offer new insights in the understanding of human implantation. The concept of implantation window is clearly challenged. Aiming to the center of the window is fine, but we still dońt know how wide is that center.

Trial registration number

not applicable
P–010 Timing of TESE does not affect laboratory outcomes
Authors: Goktas C , Basar M , Fetahovic M , Spahovic H , Goktas E , Goktolga U
Published: August 2021

Study question

What is the outcome of intracytoplasmic sperm injection (ICSI) using testicular spermatozoa obtained on the day of oocyte pick-up (OPU) or the day before OPU.

Summary answer

Testicular spermatozoa were obtaining the one day before OPU does not affect fertilization rate, top quality embryo on day 3, and blastocyst utilization rate.

What is known already

Usually, TESE is performed just before OPU. OPU is generally cancelled if no sperm is retrieved. The use of fresh testicular spermatozoa, obtained the day before OPU could offer the couple and the caring team both medical and practical advantages. The benefits of this approach, however, has not been evaluated in detail. An uncontrolled preliminary study has revealed that regular fertilization and pregnancy rates could be achieved with sperm extraction performed one day before OPU.

Study design, size, duration

This was a single-center retrospective study in Bahceci BIH IVF center. Sixty-six patients suffering from azoospermia from January 2015 to December 2020 were evaluated. TESE was performed either on the OPU day (43 patients; group A) or one day before OPU (23 patients; group B).

Participants/materials, setting, methods

In this study, primary outcomes were motile spermatozoa at ICSI, fertilization, top quality embryo on day 3, and blastocyst utilization rate. Statistical analyses were performed with chi-squared tests.

Main results and the role of chance

There is no statistical difference fertilization rate (72.3% vs. 72.2, p > 0.05), top quality embryo rate on day 3 (58.3% vs 58.3%, p > 0.05) and blast utilization rate (43.98% vs 49.58%, p > 0.05) between group A and B, respectively.

Limitations, reasons for caution

The retrospective nature of this study may not eliminate potential bias. On the contrary, the strength of our study is that all procedures were performed by the same operators, so there are no operator-dependent differences. More research is needed to prove our findings.

Wider implications of the findings: TESE procedure can be performed one day before OPU without compromising success.

Trial registration number

1
P–020 AZFc partial microdeletions of the Y chromosome is associated with severe oligozoospermia among Iranian men
Authors: AzarAfshar Z , Sadigh. Gilani M A , Ghaheri A , Zamanian M R
Published: August 2021

Study question

Are AZFc partial deletions correlated with severe oligozoospermia in Iranian men? Can we consider them as risk factors for infertility?

Summary answer

The frequency of total partial AZFc microdeletions was significantly higher in the oligozoospermia group compared to control group (8% vs. 3%, P = 0.028).

What is known already

Among many factors affecting male infertility, the second most common genetic factor is Y chromosome microdeletion. Some studies on partial AZFc microdeletions (especially on three major types; gr/gr, b1/b3 and b2/b3) have associated them with impaired spermatogenesis (azoospermia and oligozoospermia) in infertile men from different ethnicities. This finding is attributed to differences in alterations in pattern of DAZ/CDY1 copy numbers as spermatogenesis related genes.

Study design, size, duration

200 oligozoospermic (sperm count <5 mil./mL) and 200 fertile men were included as case and control groups, respectively. Individuals with karyotype abnormalities, complete microdeletions in AZF regions, infections, hypogonadism, history of chemotherapy and radiation, cryptorchidism or history of orchiopexy were not included. The study was approved by the Royan Institute Ethics Committee. Written informed consents were obtained from each participant.

Participants/materials, setting, methods

Total DNA from peripheral blood was used to amplify six sequence-tagged sites (STS) markers through multiplex PCR to detect AZFc partial deletions according to previous studies. Patterns of deletion in DAZ and CDY1 copies were determined through PCR- RFLP.

Main results and the role of chance

The frequency of AZFc partial microdeletions was 8% in oligozoospermic men (16/200) which was significantly higher compared to 3% in control group (6/200) (P = 0.028). Hence, partial deletions may be considered as a risk factor for the male infertility in Iranian population. Also, gr/gr showed a higher frequency in oligozoospermic group (4%) compared to controls (1.5%) (P = 0.126). The combination of DAZ1/2+CDY1b was the most observed deletion pattern in 8 oligozoospermic men with gr/gr deletion (75%), while among 3 controls with gr/gr, DAZ3/4+CDY1a (2 out of 3) and DAZ3/4+CDY1b (1 out of 3) were detected. Therefore, DAZ1/2+CDY1b can be correlated to oligozoospermia.

Limitations, reasons for caution

In order to achieve stronger statistical results, a larger sample size is of more help.

Wider implications of the findings: Risk of vertical transmission to male offspring and expansion in the size of deletions should be considered when providing ART services to infertile men. Genetic counseling is suggested in oligozoospermic men.

Trial registration number

-
P–039 Sperm chromatin integrity in relation to clinical pregnancy rate in an egg donation program
Authors: Vargas-Tominaga L , Suarez A , Medina A , Alarcon F , Gomez M
Published: August 2021

Study question

Is DNA Fragmentation Index (DFI) determinant in clinical pregnancy rate (CPR) after ICSI?

Summary answer

DFI is determinant in CPR after ICSI

What is known already

Male fertility evaluation often leaves the clinician in uncertainty. Semen analysis is a basic examination, but insufficient. Sperm DNA is in a compact state, and its integrity is observed to be related to reproductive capacity

Study design, size, duration

A retrospective, single center study, in 65 couples underwent egg donation, ICSI and blastocyst transfer, evaluating DFI and its effect on CPR. This study was carried out since September 2017 to March 2020

Participants/materials, setting, methods

DFI was evaluated using Sperm Chromatin Dispersion test (SCD) and considering 20% or below as normal. We performed ICSI in donated eggs, cultured until blastocyst stage and transferred 1 to 3 embryos. CPR was defined as the number of patient with fetal heart beats presents in relation with the number of patients with embryo transfer. We determined CPR in both groups with normal and abnormal DFI. Fisher exact test was used to analyze the differences.

Main results and the role of chance

From these 65 couples, in 29 male partners had normal DFI and 36 abnormal. CPR was 68.97% in the first group and 41.67% in the second (p = 0.0448)

Limitations, reasons for caution

The small number of patients is a limitation

Wider implications of the findings: The results permit us to know more male patients, to consider DFI as an important variable and to prepare better our patients for procedures.

Trial registration number

00012
P–062 Annexin-V MACS method could be a good choice for sperm selection with high PLCZ1 expression and high blastocyst rate in high DFI male factor
Authors: Saleh. Novin M , Zandie Z , Bakhtiari M , Aflatoonian R
Published: August 2021

Study question

What effect does Annexin-V MACS method have on sperm selection by pLCZ1 expression and blastocyst rate in male DNA fragmentation?

Summary answer

AnnexinV-MACS method could be a good choice for sperm selection with high pLCZ1 expression and high blastocyst rate in high DNA fragmentation male factor.

What is known already

Sperm selection based on morphology and motility in ART techniques, is not enough for choosing the best sperm especially in male factor patients. In Annexin-V magnetic activated cell sorting (MACS) technique, apoptotic sperm are separated from non-apoptotic one by negative selection. So, this method can improved quality of compaction rate in embryo. PLCζ is oocyte activating factors that it starts oscillations of calcium ca2+ in oocyte and it has a significant effect on fertilization and implantation.

Study design, size, duration

Semen samples from 30 male factor infertile couples with high DFI (DFI>30%) were selected and divided into two group (control and experimental) in each patient.

Participants/materials, setting, methods

Control was washed with DGC and experimental one was selected by MACS-DGC. Retrieved eggs in each patient, were divided in 2. Both group were injected by DGC and MACS respectively. Semen parameters and DFI (SCD test) were analyzed before and after processing. After ICSI, rate of fertilization, embryo development and blastocyst formation were evaluated. Real time PCR evaluate expression of PLCζ. Comparison between results of two groups was assessed by SPSS analysis.

Main results and the role of chance

Results showed that, sperm motility and morphology after MACS method (45%, 1.7%) was significantly higher than DGC method (40%, 1.1%) and before washing (35%, 0.9%). Percent of DFI in MACS group (36%) was significantly decreased compared to DGC (45%) and primitive group (55%). The number of oocytes were injected in DGC group was 93 and in MACS group was 111. Fertilization rate in both groups was almost the same (72.07% in MACS vs 73.11 in DGC). Rate of day 3 embryo with good grade in MACS group (72. 5%) was significantly higher than DGC (51.47%) (P < 0.05). The blastocyst rate in MACS-DGC group (69.69%) was significantly increased compared to DGC group (48%). PLCζ gene expression in MACS-DGC was significantly higher than DGC group (p-value=0.046).

Limitations, reasons for caution

This experiment was performed invivo.

Wider implications of the findings: Sperm selection by MACS-DGC method can improve sperm motility, morphology and reduce sperm DFI. No significant difference was observed in fertilization rate, but percent of high-quality embryo on days 3 and 5 was significantly higher by this method,also it can be suggested as a good choice for patients with highDF.

Trial registration number

Not applicable
P–067 Utility of evaluating semen samples from adolescents with Klinefelter Syndrome for cryopreservation: A multi-institution evaluation
Authors: Chu K , Punjani N , Nassau D , Kashanian J , Ramasamy R
Published: August 2021

Study question

Should physicians continue to evaluate semen analysis from adolescents with Klinefelter Syndrome for fertility preservation?

Summary answer

In the largest multi-institutional retrospective database to-date for this patient population, no sperm was found in ejaculate for cryopreservation amongst adolescent males with Klinefelter Syndrome.

What is known already

Klinefelter Syndrome is the most common genetic condition leading to male infertility and non-obstructive azoospermia. The condition causes decreased testicular growth, leading to lower production of testosterone and resulting deficiencies in secondary sexual characteristics. While testosterone therapy may be required for hypogonadism, there may be impact on future fertility potential. Current practice is to have KS adolescent patients provide semen analyses to identify potential sperm for cryopreservation. While the incidence is low, current epidemiological studies have been with limited sample size.

Study design, size, duration

This was a retrospective study of all adolescent Klinefelter Syndrome patients seen at the male infertility clinics of two large academic institutions between the years of 2015 to 2020. Adolescence was defined as the ages of 10 – 19 years old, as per the World Health Organization.

Participants/materials, setting, methods

A total of 116 patients were identified for the retrospective study database. Demographic information including weight, height, comorbidities, concurrent medications were collected. Hormone levels such as FSH, LH, testosterone, and estrogen were included for 77 patients. Additionally, semen analyses were available for 49 patients. Main results and the role of chance: Of the 49 patients with semen analyses, only 3 patients had rare sperm in ejaculate not sufficient for cryopreservation while the remaining had azoospermia. The average ejaculate volume of the provided semen samples was 0.9 cc. The average serum total testosterone level of adolescent Klinefelter Syndrome patients was 236 ng/dL. As expected, gonadotropin levels were found to be elevated (mean: 18.47 IU/L for FSH and 9.12 IU/L for LH).

Limitations, reasons for caution

The main limitation for this study was the sample size.

Wider implications of the findings: The findings from the largest retrospective study of this patient population imply a need to revisit counseling regarding the need for semen analyses in adolescent Klinefelter Syndrome patients.

Trial registration number

Not applicable
P–071 Human Papillomavirus Infection and impact on men fertility
Authors: Madzunkov N , Madjunkova K
Published: August 2021

Study question

Is there any correlation between men infertility and HPV infection and its impairment on sperm quality?

Summary answer

There is a significantly higher prevalence of high-risk HPV in infertile men than fertile men. HPV infection does not impair sperm quality.

What is known already

Many factors may cause the infertility in males and females. Human papillomavirus are the most frequently sexually transmitted DNA viruses and etiological agents of cervical cancers. There is association between HPV infection in females and adverse pregnancy outcomes such as spontaneous abortion and spontaneous preterm delivery. Previous studies have reported the detection of HPV DNA in semen and in different sites of the male reproductive tract, such as glans penis and scrotum. Recent reports suggested that HPV may affect sperm parameters and lead to male infertility. The impact of HPV infection upon male fertility abnormality has received far less attention.

Study design, size, duration

In this study case control study we examined 38 fertile and 36 men from infertle couples.

Participants/materials, setting, methods

we examined the swabs of the entire penile surface and semen samples for HPV detection and genotyping from 38 fertile men and 36 from men from infertile couples.HPV were detected with PCR method. Sperm was also examined for its motility, sperm quantity and morphology .

Main results and the role of chance

Among 36 confirmed infertile males, only 8 (22.22%) cases were tested positive for HPV of semen samples and 2 among fertile men were HPV-positive (5.26%) of semen samples. Among infertile males 14 (38.88%) had HPV positive penile swabs, and only 1 (2,63%) had positive HPV swab among fertile men. The most prevalent HPV types in the male external genitalia were HPV–16. The most prevalent HPV types in semen were HPV–53. This data revealed a significant association between high-risk HPV and male infertility (P = 0.03). Sperm quality (morphology and motility) did not differ significantly between men with seminal HPV infection and uninfected men.

Limitations, reasons for caution

There were some limitations in the study such as differences in age. sample sizes and the number of HPV genotypes detected.

Wider implications of the findings: We need larger studies and more further investigations to confirm the impact of HPV on male infertility.

Trial registration number

2
P–077 Performance of the postwash total motile sperm count as a predictor of pregnancy at the time of intrauterine insemination
Authors: Kasturiraj A , Reddy S , Daniel M , Namboor. Srinivasan S , Raja N , Reddy E
Published: August 2021

Study question

Is the performance of the postwash total motile sperm count a predictor of pregnancy at the time of intrauterine insemination?

Summary answer

The number of motile spermatozoa inseminated (NMSI) in IUI can be used to define clear range of pre /postwash sperm parameters.

What is known already

There is no consensus about the optimal number of motile spermatozoa inseminated (NMSI) required for a reasonable chance of pregnancy after IUI. A meta-analysis of 16 studies assessing NMSI and IUI outcomes, concluded that at cut-off levels between 0.8 and 5 million, defined as the ability to predict failure to become pregnant. The purpose of this study is to determine the range of NMSI as a predictor of success in IUI.

Study design, size, duration

This prospective study includes 60 patients who underwent semen analysis at an academic infertility centre (SRIHER) during the month of December 2020 and January 2021.

Participants/materials, setting, methods

A total of 60 infertile couples who underwent IUI at our academic centre were enrolled in our study. A detailed history and infertility work up was done before proceeding for IUI, as per the department protocol. The semen was prepared by discontinuous 2 layered density gradient method. The results were analyzed by patient factors including age, BMI, semen parameters, NMSI. The NMSI were divided into 4 groups: A (<1), B (1–4), C (5–9), D (>10).

Main results and the role of chance

The mean age of the infertile couples who underwent IUI was (28.2 ± 3.8) in females and (31.8 ± 3.8) in males respectively. The sperm parameters such as concentration (21.8 ± 14.8), motility (53.15 ± 13.22), morphology (2.43 ± 1.33) respectively. When the NMSI was group C (5–9 x 10 6) the pregnancy rate was 38.5% whereas the pregnancy rate was 37.5% in group D ( >10 x 10 6). In the other sub groups such as group A(<1 x 10 6) and group C (1–4 x 10 6) the pregnancy rate was 14.2% and 12.5% respectively.

Limitations, reasons for caution

Infertile men with Azoospermia, Men with Retrograde ejaculation, Testicular samples, Epididymal samples, Infected samples. All of the above samples were avoided. It cannot be used for counselling during the initial infertility workup, but only during/after the IUI procedure.

Wider implications of the findings: The results suggest that NMSI can be a predictor of success in IUI in patients who are < 30 years of age & ≥35 years, NMSI does not appear to be a useful. The effect of NMSI on pregnancy rate needs to evaluated on a larger scale .

Trial registration number

Not applicable
P–102 GM-CSF (granulocyte-macrophage colony-stimulating factor) as a sperm medium supplement improves sperm quality in Oligoastenoteratospermia (OAT) men by activating the PI3K/Akt pathway
Authors: Tanhay. Kalat. Sabz F , Ashrafi M , Amjadi F , Zandieh Z , Hosseini E , Aflatoonian R
Published: August 2021

Study question

Can GM-CSF as a sperm medium supplement improve sperm quality in OAT men?

Summary answer

GM-CSF can be used to improve sperm quality in OAT men by activating the PI3K/Akt pathway.

What is known already

OAT patients have very low sperm parameters, including morphology, count, and motility, which can adversely affect the results of assisted reproduction technologies. It seems the development of sperm media is necessary to improve the sperm parameters of these patients. GM-CSF is a natural growth factor produced by the reproductive organs, culture media supplemented with GM-CSF is widely commercially available and enhances the embryo development and implantation rate. Studies show that this growth factor in the semen of infertile men is lower than that of fertile men. However, there is no study to assess the effect of GM-CSF on sperm quality.

Study design, size, duration

In the present study, Semen specimens were collected from 20 OAT patients who have male infertility factors, according to WHO criteria. After the swim-up washing procedure, each of the samples is divided into two groups; experiment, and control. In the experimental group, samples are incubated with a medium containing 2 ng/ml GM-CSF for one hour, yet, in the control group, the sperms are incubated without GM-CSF for the same time.

Participants/materials, setting, methods

The sperm motility was examined with phase-contrast microscopy, Eosin-nigrosin staining method was used to assess sperm viability. The expression of sperm glucose transporters (GLUT 1, 3) was determined using immunofluorescent staining, the ratio of pAkt to total Akt was assessed by the Western blotting method. Image J software was used to quantify results; the data was analyzed by SPSS software. P-value<0.05 was considered statistically significant.

Main results and the role of chance

As compared to the control group, supplementation with GM-CSF improved sperm progressive motility, enhanced GLUT 1 and 3, and p-AKT/AKT expression (P < 0.05). There was no significant difference between the viability of the control and experimental groups.

Limitations, reasons for caution

This study needs further investigations, including Real-time PCR for the genes of this signaling pathway which is ongoing.

Wider implications of the findings: We showed for the first time that GM-CSF can improve sperm quality by influencing motility and energy metabolism in spermatozoa which can be affected by increasing the phosphorylation of AKT. This growth factor could be an appropriate supplement in sperm media for OAT patients.

Trial registration number

IRCT20200519047508N1
P–103 Novel sperm preparation techniques compared with conventional preparation method
Authors: Vahidi N , Amjadi F S , Kalat. sabz F , Zandie Z , Narimani N
Published: August 2021

Study question

Which sperm preparation technique separate the best quality sperm?

Summary answer

Microfluidic method improved the sperm parameters and decreased sperm DNA damage.

What is known already

About 40% of infertility issues are due to male factor. One of the known causes of male infertility is associated with low sperm parameters and high level DNA fragmentation. Sperm preparation techniques in ICSI procedures is used in order to obtain the best-quality sperm.

Study design, size, duration

The present study was designed to compare Microfluidic, Zeta potential, Magnetic Activated Cell Sorting (MACS) and Swim-up methods for sperm preparation and the effect of these methods on semen parameters and sperm DNA integrity in infertile men (n = 25) with a mean age of 38.

Participants/materials, setting, methods

In this study, each sample was divided into 4 groups, one part for preparing by Microfluidic method, one of them for preparing by Swim -up method, the other one was prepared by MACS and the last one was prepared by zeta potential. Then sperm count, viability, motility and morphology were assessed according to WHO 2010. DNA damage were assessed by Sperm DNA Fragmentation assay and sperm chromatin packaging assessed by CMA3 staining test

Main results and the role of chance

Sperm parameters including viability, motility, and morphology in the Microfluidic method were significantly improved and sperm DNA damage were significantly lower than three other methods (P-value <0.05). The sperm parameters and sperm DNA damage after preparation by MACS and Zeta potential methods were not significantly different however in the Swim-up method sperm parameters were lower than three other methods (P-value <0.05).

Limitations, reasons for caution

The fertilization and pregnancy rate of the resulting embryos are not available.

Wider implications of the findings: Our results showed that Microfluidic can be an effective way to improve sperm quality of infertile male compared to conventional preparation methods. We also found instead of the MACS method, we can use the Zeta potential method according to their costs, for sperm preparation during ART cycle.

Trial registration number

*