bluuebunny/crossref_metadata_2025 huggingface dataset:
P–007 Effect of swim-up on sperm morphology
Authors: Ganeva R , Parvanov D , Handzhiyska M , Stamenov G
Published: August 2021

Study question

To evaluate the effect of swim-up on the percentage of certain morphological defects in the semen population

Summary answer

Swim-up preparation led to significantly lower percentage of spermatozoa with cytoplasmic droplets, thick neck and also multiple defects.

What is known already

Swim-up is routinely used sperm preparation technique in ART practice. It is widely known that swim-up enhances sperm quality in terms of motility and sperm morphology. However, the effect of swim-up on the frequency of occurrence of the specific sperm morphological abnormalities is still missing.

Study design, size, duration

This observational study involved 30 teratozoospermic patients of Nadezhda Women’s Health hospital between December 2020 and January 2021. Sperm morphology was evaluated before and after swim-up preparation.

Participants/materials, setting, methods

Native semen was liquefied and was subjected to swim up. Semen analysis performed according to WHO 2010. Native semen and swim up samples from the same men were subjected to Kruger strict morphological evaluation.

The analyzed sperm morphological defects included: head defects (large, small, tapered, pyriform, round, amorphous and double heads); midpiece defects (bent, asymmetrical, thin, thick, presence of cytoplasmic droplet); tail defects (short, hairpin, bent, coiled tail and terminal droplet) and multiple defects.

Main results and the role of chance

Wilcoxon paired test showed that the percentage of morphologically normal spermatozoa was significantly higher in the swim-up samples in comparison to the native semen (8.5±4.2% vs 4.9±3.2%, p < 0.05). In addition, the percentage of spermatozoa bearing multiple defects was found to be significantly lower in the swim-up samples than in the native semen (25.8±11.6% vs 37.0±15.0%, p < 0.05). Two specific sperm morphological defects were found to be significantly lower after swim-up preparation: the presence of cytoplasmic droplets (6.0±1.0% vs 8.6±1.5%, p < 0.05) and the thick neck (9.7±5.5% vs 12.8±5.8%, p < 0.05). No significant different were observed in the other morphological defects between swim up samples and native semen (p > 0.05).

Limitations, reasons for caution

Results obtained from this study need to be confirmed by larger group of samples.

Wider implications of the findings: Our study showed a significant reduction of certain midpiece defects after swim-up. The observed selection of spermatozoa without thick necks and cytoplasmic droplets explains the effectiveness of swim-up on ART. In addition, the obtained results can serve as a guide for future validation of new sperm preparation techniques.

Trial registration number

Not applicable
P–011 Automated sperm morphology assessment using artificial intelligence technology
Authors: Agarwal A , Panne. Selvam M K
Published: August 2021

Study question

Can LensHooke X1 PRO semen analyzer be used to evaluate sperm morphology in men with infertility?

Summary answer

Morphology results generated by X1 PRO are highly reliable when normal sperm forms are ≥4% and therefore they can be reported in such cases .

What is known already

Most laboratories rely on manual evaluation of sperm morphology smears, which is a time-consuming procedure and its results are subjected to a relatively high variability. However, in recent years the computer-assisted semen analyzers are being increasingly used to evaluate sperm morphology. The X1 PRO semen quality analyzer was designed for in vitro diagnostic use to analyze sperm concentration, total, progressive and non-progressive motility as well as sperm morphology based on WHO 5th edition criteria. Evaluation of sperm morphology using X1 PRO based on AIOM (Artificial Intelligence Optical Microscopic)-based technology requires no fixation steps or staining unlike the manual method.

Study design, size, duration

This cross-sectional study used 31 semen samples from 8 normozoospermic healthy volunteers and 5 infertile men with a minimum abstinence period between 2 - 3 days. While the 8 healthy semen donors produced a total of 26 ejaculates, which were split into 88 aliquots, the 5 infertile patients produced 5 ejaculates that were split into 13 aliquots.

Participants/materials, setting, methods

A total of 101 aliquots were prepared from the native semen samples either by dilution or concentration using seminal plasma of the respective donors. Automated semen analysis was performed by the X1 PRO semen analyzer and the results of sperm morphology were compared with manual morphology results using Diff-Quik staining. Statistical analysis was carried out to calculate the positive predictive value (PPV) and negative predictive value (NPV) of X1 PRO semen analyzer.

Main results and the role of chance

The X1 PRO sperm morphology results show a weak non-significant (P = 0.2441) correlation (r = 0.119) with the manual results. However, X1 PRO demonstrated a high PPV (97.7%) and a low NPV (9.1%) for correct assessment of sperm morphology (≥4%) when compared to manual results. Due to its high PPV, laboratories can report the morphology results generated by X1 PRO in all such cases when normal sperm forms are ≥4%. However, a manual evaluation is necessary in patients with abnormal morphology (<4%).

Limitations, reasons for caution

One of the limitation of this study is that X1 PRO morphology values did not correlate with manual results. The low NPV seen in our study is due to the inclusion of very few samples with abnormal sperm forms (<4%) in the analysis.

Wider implications of the findings: The X1 PRO’s combination of speed, ease of use, accuracy and portability makes it a good choice of device for small medical offices to large IVF centers. High PPV of X1 PRO allows it to correctly identify normal sperm forms for diagnostic use.

Trial registration number

18–771
P–027 Does air pollution impact on semen parameters? Findings from a real-life cross-sectional study in white-European infertile men
Authors: Belladelli F , Pozzi E , Boeri L , Fallara G , Capogrosso P , Cignoli D , Candela L , Schifano N , Raffo M , Colandrea A , Montorsi F , Salonia A
Published: August 2021

Study question

We aimed to investigate the association between air pollutants levels and semen parameters in a cohort of white-European men seeking medical attention for couple’s infertility.

Summary answer

We found that Pm10, Pm 2.5, and NO2 levels were negatively associated with sperm morphology.

What is known already

Air pollutants levels have been monitored closely for environmental and research issues in industrialized countries.

Study design, size, duration

Data from 156 infertile men consecutively assessed between 01/2019 and 12/2020 were analysed.

Participants/materials, setting, methods

Semen analyses were based on 2010 WHO reference criteria. We analysed the annual average level of the main markers of air pollution (Pm10, Pm2.5, and NO2) between 2014–2018 (Legambiente, 2020, Annual dossier series on air quality in Italy) relative to patients’ addresses of the last 5 years. Health-significant comorbidities were scored with the Charlson Comorbidity Index (CCI). Descriptive statistics and linear regression analyses were used to test the association between air pollutants and semen parameters.

Main results and the role of chance

Overall, median (IQR) age and BMI were 37 (33–41) years and 25.1 (23.4–27.3) kg/m2, respectively. A CCI≥1 was found in 12 (7.7%) men, and 47 (30.1%) participants were smokers. As for sperm parameters, semen volume, sperm concentration, sperm progressive motility and normal sperm morphology were 3 (2–4) ml, 18 (5–45.5) x106/ml, 32 (22–45)% and 2 (1–3)%, respectively. Pm10 was negatively associated with sperm morphology (ß: –0.580, p = 0.001). Furthermore, Pm10 was found to be an independent predictor for sperm morphology worsening (ß: –0.729, p = 0.006) (Fig.1A), after adjusting for age, BMI, CCI and smoking status. Similarly, Pm2.5 levels were negatively associated with sperm morphology (ß: –0.595, p = 0.009) (Fig.1B). After adjusting for the same variables, the association between Pm2 and sperm morphology remained significant (ß: –0.724, p = 0.015) (Fig.1C). NO2 levels were found to be associated with sperm morphology worsening after adjusting for age, BMI, CCI, and smoking status (ß: –0.471, p = 0.006).

Limitations, reasons for caution

It is a retrospective analysis at a single, tertiary-referral academic centre, thus raising the possibility of selection biases. Moreover, markers of air pollutions divided by residence may not completely evaluated the single patient exposure.

Wider implications of the findings: In this cross-sectional study we found that Pm10, Pm 2.5, and NO2 levels were negatively associated with sperm morphology, while they were not consistently associated with an increased risk of other abnormal sperm parameters in infertile men. Further studies are needed to characterize air pollution effects on sperm parameters.

Trial registration number

Not applicable
P–044 A non-classical splice site variant in ANOS1 gene leading to Kallmann syndrome
Authors: Guo H , Xia Y , Cui C
Published: August 2021

Study question

Genetic risk of the non-classical splice site variant in ANOS1 gene

Summary answer

A non-classical ANOS1 splice site variant, c.1062 + 4T>C, causes Kallmann syndrome.

What is known already

Genetic abnormalities play a key role in the development of Kallmann syndrome. Although an overwhelming majority of missense and nonsense mutations occur in the exons of a gene, intron mutations can also be pathogenic.

Study design, size, duration

The research object is a family. Eight patients of the family were recruited in this study, three of them were diagnosed with Kallmann syndrome.

Participants/materials, setting, methods

Genomic DNA was extracted from peripheral blood and whole-exome sequencing (WES) was performed to identify the genetic abnormalities. PCR was performed to verify the WES results. The functional splicing reporter mini gene assay was performed to assess the impact of sequence variants on splicing.

Main results and the role of chance

The proband and other two patients exhibited the typical clinical features of KS. A non-classical splice site variant, c.1062 + 4T>C in ANOS1 gene was identified, whereas the other unaffected family members did not have this mutation. This mutation caused the disappearance of the splicing site of intron 7 and the splicing position became the 156th base of exon 7, which caused a frame-shift mutation, leading to a premature termination of translation.

Limitations, reasons for caution

Since the ANOS1 gene is almost not expressed in the blood, in order to uncover the effect of this splice site variant of ANOS1, we carried out a functional splicing reporter mini gene assay in the mini gene vector pEGFP-N1.

Wider implications of the findings: This study shows that mutations in non-classical splicing regions are also pathogenic. Therefore, it is recommended that the detection and analysis of this gene should pay attention to the non-classical splice site variant.

Trial registration number

Not applicable
P–069 microfluidic sperm sorting vs density gradient to yield sperm with reduced DFI for patients undergoing IVF-ICSI
Authors: Makwana D P , Makwana S , Sen T
Published: August 2021

Study question

To compare the effect of sperm preparation methods on the DFI of semen sample for patients undergoing ICSI.

Summary answer

On comparing the results, microfluidic sperm sorting yielded sperms with significantly less DFI as compared to density gradient method of sperm preparation.

What is known already

The DNA integrity of the sperm plays an important role to ensure formation of good quality embryos with increased potential of fertilization, growth and ultimately implantation.. Centrifugation has shown to add stress to the sperm and leading to DNA damage, therefore there is a need to develop techniques of sperm preparation which help in retrieving as many sperms with intact DNA from the unprocessed sample as possible. Microfludic is fluid dynamic based technique of sperm preparation. in this study, we evaluated if microfluidic sperm sorter can recover motile sperm with better DNA integrity compared to density gradient preparation method.

Study design, size, duration

Prospective randomized study conducted in 80 patients undergoing IVF-ICSI with normal semen parameters (based WHO criteria 2010). DFI was done using Sperm Chromatin Dispersion (SCD) test in split semen samples prepared by microfluidic sperm sorter and density gradient method. Sperm morphology and motility were also recorded and evaluated based on the WHO 2010 criteria.

Participants/materials, setting, methods

Semen parameters of the sample were assessed by microscopic examination. DFI of each unprocessed sample was carried out using SCD test, following that the sample was split and sperm preparation was done using microfluidic sperm sorter and density gradient. the recovered sperm were tested for DFI and the results were compared.

Main results and the role of chance

Mean DFI in unprocessed semen samples was 23%. the analysis of split semen samples post preparation showed that the DFI was significantly reduced with the use of microfluidic sperm sorter (mean DFI 0.6%) as compared to density gradient (mean DFI 9%).

Limitations, reasons for caution

A major limitation of the microfluidic sperm sorter is the use sperm concentration and motility of the semen sample. In oligospermic and asthenospermic samples, density gradient is the preferred method of preparation. Lack of data showing improvement in clinical outcomes with reduced DFI is also a major limitation.

Wider implications of the findings: Microfluidics has shown to significantly reduce the DFI of the semen sample, it requires no extra equipment and cost and is relatively easy to pick up. Density gradient method of sperm preparation continues to be the preferred method due to its versatility and recovery of good quality sperm.

Trial registration number

Not applicable
P–070 Evaluation of SARS-CoV–2 in human semen and effect on total sperm number: A prospective observational study
Authors: Best J , Kuchakulla M , Khodamoradi K , Lima T , Frech F , Achua J , Rosete O , Mora B , Arora H , Ibrahim E , Ramasamy R
Published: August 2021

Study question

Is the SARS-CoV–2 virus present in human semen and what is the impact on semen parameters following an infection?

Summary answer

SARS-CoV–2 infection, though not detected in semen of recovered men, can affect TSN in ejaculate in the acute setting.

What is known already

Early epidemiological data has suggested that the primary mode of transmission is through respiratory droplets, but the presence of SARS-CoV–2 has been identified in other bodily fluids such as feces, urine, and semen.

Study design, size, duration

We prospectively recruited thirty men diagnosed with acute SARS-CoV–2 infection using real-time reverse transcriptase-polymerase chain reaction (RT-PCR) of pharyngeal swab specimens. Thirty semen samples from recovered men were obtained 11–64 days after testing positive for SAR-CoV–2 infection. The median duration between positive SAR-CoV–2 test and semen collection was 37 days (IQR=23).

Participants/materials, setting, methods

Semen samples were collected from each individual using mailed kits. Follow-up semen samples were done with mailed kits or in-person in office setting. Semen analysis and PCR was performed after samples were received.

Main results and the role of chance

The median total sperm number (TSN) in ejaculate was 12.5 million (IQR=53.1). When compared with age-matched SARS-CoV–2(-) men, TSN was lower among SARS-CoV–2(+) men (p = 0.0024). Five men completed a follow-up sperm analysis (median 3 months) and had a median TSN of 18 million (IQR=21.6). No RNA was detected by means of RT-PCR in the semen in 16 samples tested.

Limitations, reasons for caution

First, most of the semen samples came from non-severe men of whom were in the recovery stage and lacked symptoms. Additionally, our sample size was relatively small and overnight mail-in semen analysis kits were used during the acute phase of infection to minimize contact with positive subjects.

Wider implications of the findings: Our findings suggest extremely low risk of viral transmission during sexual contact and assisted reproductive techniques, although further data need to be obtained. The impact on TSC in recovered men from SARS-CoV–2 infection is concerning, nevertheless long-term follow-up of these men is critical to determine the nadir of TSC.

Trial registration number

20200401
P–082 Effect of semen hyper viscosity (SHV) on blastocyst formation rate and implantation rate
Authors: Suthar A , Sharma N , Mishra V , Aggarwal R , Sheth H , Patel K
Published: August 2021

Study question

Does semen hyper viscosity effects blastocyst formation rate

Summary answer

Hyper viscosity of semen sample later results in poor blastocyst formation rate and lower implantation rate.

What is known already

Normal range of semen hyper viscosity ranges between 12–29%.Highly viscous semen samples impairs the physical and chemical characteristics of seminal fluid and due to which seminal oxidative damage increases which further increases the ROS and reduces the sperm motility there are some factors that can affect the seminal viscosity out of which one is Male accessory gland infection, Hypo function of prostate seminal vesicles and varicoceles. SHV create hindrance in semen preparation.

Study design, size, duration

Retrospective study was conducted from June 2019 to Oct 2020 at IVF unit IKDRC hospital.

Participants/materials, setting, methods

142 patients were enrolled from June 2019 to Oct 2020 in IVF unit IKDRC hospital and divided into two groups. Group A (n = 83) patients with hyper semen viscosity and Group B (n = 69) patients with normal semen viscosity, inclusion and exclusion criteria’s were same for both the groups, only patient with normozoospermia were taken. Semen analysis was done by using WHO manual 2010.

Main results and the role of chance

In group A with hyper semen viscosity fertilization rate was (49.2% vs. 70% p = <0.001) vs in group B with normal semen viscosity which is significantly higher in group B, Blastocyst formation rate ( 18.4% vs 35% p = <0.01) and implantation rate (9.4% vs 20% p = <0.005) both are significantly higher in group B . Which implies fertilization rate , blastocyst formation rate and implantation rate is significantly lower in patients with semen hyper viscosity.

Limitations, reasons for caution

Larger randomized control studies are needed to strengthen these results.

Wider implications of the findings: Our study demonstrates that patients having higher semen viscosity have poor blastocyst formation rate and implantation rate due to oxidative stress.

Trial registration number

Not applicable
P–085 Sperm retrieval after microdissection testicular sperm extraction (micro-TESE) and pregnancy outcomes of intracytoplasmic sperm injection (ICSI) in men with Klinefelter syndrome
Authors: Dinh V H , Nguye T H H , H. Nguye B , T. Nguye A , C. Trin K , Q. H D , D. Nguye M , H. L T , H. Pha M
Published: August 2021

Study question

What are the sperm retrieval rate and ICSI outcomes in azoospermic men with Klinefelter syndrome (KS)?

Summary answer

In men with KS, a sperm retrieval rate of 51.3% after the first attempt micro-TESE, and 4 live births after ICSI were observed. What is known already: Klinefelter syndrome (KS) is encountered in 10% of men with azoospermia. Micro-TESE is presently used to treat infertility for KS patients with nonobstructive azoospermia. The retrieved sperms can be used for ICSI. Study design, size, duration: From June 2019 to July 2020, 39 azoospermic patients with KS were examined for the presence of testicular spermatozoa. Participants/materials, setting, methods: Participants were recruited from couples attending the Andrology and Fertility Hospital of Hanoi Vietnam, for infertility treatment. Micro-TESE was performed to extract testicular tissue. After retrieval, ICSI was used with fresh sperm. Main results and the role of chance: The sperm retrieval rate of first attempt micro-TESE in KS men was 51.3% (20/39). Logistic regression analysis showed patient age did not affect the sperm retrieval rate of micro-TESE (OR 0.99, 95% CI 0.88 - 1.11). Similarly, no association was observed between serum FSH, LH, testosterone level, and testicular volume with the success of sperm retrieval. The fertilization rate after ISCI in patients with retrieved sperm was 60% (12/20). Clinical pregnancy and ongoing pregnancy rates were 50% (10/20) and 40% (8/20). There were 4 live births. No sufficient data were available to test the effect of clinical or biological parameters on ICSI outcomes.

Limitations, reasons for caution

Our data rely on a cohort of KS patients attending a single fertility clinic. The sample size did not allow regression analysis for any ICSI outcome.

Wider implications of the findings: Micro-TESE is helpful in retrieving sperm in azoospermia KS patients with sperm retrieval rate reaching 50%. ICSI following micro-TESE can lead to an ongoing pregnancy rate of 40% with some result in live births. The outcome of micro-TESE is independent of any clinical or biochemical parameters tested.

Trial registration number

Not applicable
P–089 Paternal age affects the performance of EAU guidelines for genetic testing in infertile men: implications for candidate selection
Authors: Ventimiglia E , Boeri L , Pozzi E , Belladelli F , Fallara G , Candela L , Capogrosso P , Schifano N , Cignoli D , Colandrea G , Cornelius J , Mattei A , Abbate C , Montorsi F , Salonia A
Published: August 2021

Study question

We aimed at challenge EAU Guidelines for genetic testing performance in infertile men according to normal vs. advanced paternal age (APA).

Summary answer

EAU Guidelines perform better in karyotype analysis (KA) and Y-chromosome microdeletions (YCM) investigation in men above 35 years of age.

What is known already

EAU Guidelines for genetic testing in infertile men recommend specific threshold for candidate selection for both KA (<10 million spermatozoa/ml) and YCM (<5 million spermatozoa/ml). However, paternal age is not taken into account for candidate selection in this setting.

Study design, size, duration

Data from 2188 infertile men (according to WHO definition) consecutively evaluated at a single academic centre were analysed.

Participants/materials, setting, methods

Demographic, clinical and laboratory data were analysed. Semen analyses were based on 2010 WHO criteria. Advanced age was defined as above 35. All men underwent KA and YCM testing. EAU Guidelines were validated in our cohort and according to APA. Specificity, sensitivity, and AUC were estimated for all scenarios. A Wald-type test compared AUC according to APA for KA and YCM. Decision curve analysis (DCA) estimated the benefit of using EAU Guidelines according to APA.

Main results and the role of chance

Median (IQR) paternal age was 37 (34–41) years. Advanced paternal age was found in 1306 (60%) of included men. Prevalence of KA and YCM was 4% (48 men) and 1% (13 men), respectively. EAU Guidelines sensitivity, specificity and AUC in the overall population were 85%, 47% and 66% for KA, whereas they were 100%, 57% and 80% for YCM. When stratifying according to APA, EAU Guidelines performed better in men over 35 both in terms of KA and YCM detection. Specifically, AUC for KA detection in men with APA was 70% vs. 63% in younger men (p = 0.04). AUC for YCM detection in men with APA was 82% vs. 79% in younger men (p = 0.03). DCA confirmed higher net benefit in using EAU Guidelines in old vs. young men for the detection of both KA and YCM.

Limitations, reasons for caution

It is a retrospective analysis at a single, tertiary-referral academic centre, thus raising the possibility of selection biases.

Wider implications of the findings: EAU Guidelines for genetic testing in infertile men perform differentially according to APA. KA and YCM are better detected in older men, likely due to a wider pool of confounding etiological factors in young men. These results suggest the implementation of more accurate predictive models in younger men.

Trial registration number

Not applicable
P–099 MYD88 dependent pathway through TLR 1,2 and 6 activation play a role in interaction of high DNA fragmented human sperm with fallopian tube epithelial cells
Authors: Govahi A , Zahra Z , Fatemehsadat A , Azin A , Reza A
Published: August 2021

Study question

How does sperm with high DFI disrupt the female reproductive system?

Summary answer

Sperm with damaged DNA could disrupt the female reproductive system by production of high inflammatory factors through activation of TLRs and MyD88-dependent pathway .

What is known already

Previous studies have also shown that the interaction between sperm and fallopian tube is a complex process and activates a number of immune mechanisms. Many TLRs, including TLR1, TLR2, and TLR6 are expressed in fallopian tube epithelial cells. TLRs have an important role in the immune interaction between fallopian tube and sperm. They are also involved in sperm capacitation, fertilization and pregnancy. Examining this interaction may lead us to valuable data.

Study design, size, duration

In this study, 10 Recurrent Implantation failure couples with high DFI ( >30% by SCSA evaluation) and 10 healthy donors men with low DFI (<30%) were considered as high DFI group and low DFI group respectively. After fresh semen preparation, sperm were co-cultured with human fallopian tube epithelial cell line( OE-E6/E7) for 24 hours.

Participants/materials, setting, methods

RNA were extracted from the cell line and PCR array for human innate and adaptive immune responses was performed by RT2 profiler PCR array.

Main results and the role of chance

Analysis of PCR array data showed that the expression of TLR–1, TLR–2, TLR–6, MYD88, TIRAP, IRAKS,TRAF6, MAPKS, NF-KB, G-CSF, GM-CSF, CXCL8, CXCL10, CCL2, IL–6, IL–1, TNFa in high DFI group was significantly higher than control group. All these factors are involved in the MyD88-dependent pathway. Our research suggests that the MyD88-dependent pathway through TLR1,2 and 6 activation may be the main inflammatory pathway activated by sperm with high DFI. The released DAMPs through damaged sperms were recognized by TLRs on the epithelial cells. Following the TLR1,2 and 6 signaling, and MYD88 activation, epithelial cells were producing the inflammatory cytokines which result in the neutrophils infiltration. In the lumen of the fallopian tube, the neutrophils were activated via TLR1/TLR2, complexes that lead to the neutrophils activation, phagocytosis, NET formation, and apoptosis.

Limitations, reasons for caution

Studying with a larger sample size and examining the final factors at the protein level will be better to detect the effect of sperm DNA fragmentation on fallopian tube.

Wider implications of the findings: MyD88-dependent pathway could be one of the mechanisms that involved in interaction of high DNA fragmented sperm with female reproductive tract.

Trial registration number

Not applicable
P–112 Association between dietary total antioxidant capacity and semen quality parameters in male partners of couples attempting fertility
Authors: Eslamian G , Rohani S , Shoaibinobarian N
Published: August 2021

Study question

Is there any association between dietary total antioxidant capacity (TAC) and semen quality parameters in male partners of couples attempting fertility?

Summary answer

Greater adherence to diets high in TAC was significantly associated with higher total sperm count, sperm concentration, and sperm motility.

What is known already

Among multiple factors that affect the etiology of poor semen quality and male infertility, dietary factors have an important contribution. Also, chronic oxidative stress negatively effects semen quality. Whether adherence to the diet rich in antioxidants is associated with better semen quality remains largely unexplored.The concentration of single antioxidant cannot show the total antioxidant power of the diet, therefore the concept of dietary total antioxidant capacity (TAC) was invented.Dietary scores are useful approach to evaluate the degree of adherence to specific dietary pattern and its benefits in regard to health

Study design, size, duration

This was a cross-sectional study of 350 men from couples attending a fertility center in Tehran, Iran, recruited between June 2015 and September 2019. Men aged 25–50 years with complete dietary data were analyzed.

Participants/materials, setting, methods

Diet was assessed via a reproducible and valid 168-item semi-quantitative food frequency questionnaire to determine the entire antioxidants of the usual diet in order to calculate dietary TAC. Semen quality was assessed according to World Health Organization 2010 guidelines. The association between dietary TAC and semen parameters were assessed adjusting for potential confounders through multiple logistic regression analysis.

Main results and the role of chance

The average age of study participants was 34.9 ± 7.6 years and their BMI was 28.5 ± 4.3 kg/m2. Participants in the lower tertile of the dietary TAC were lower educated (p < 0.01), more physically active (p < 0.05), and predominantly had abnormal sperm progressive motility (p < 0.01). In the multivariable adjusted models, men in the lowest tertile of the dietary TAC Score had 2.9 times higher likelihood of having abnormal sperm motility, total sperm count and concentration, compared to men in the highest tertile of the TAC score (p < 0.05).

Limitations, reasons for caution

The main limitation of our study was its cross-sectional design, limiting our ability to derive causal association.

Wider implications of the findings: Our study suggests that dietary pattern comprising mainly of antioxidant nutrients may help improve semen quality. Our results are consistent with previous studies showing that plant-based diet contains higher levels of antioxidants are associated with better measures of semen quality.

Trial registration number

Not applicable
P–114 Correlation of GSTM1 polymorphism and oxidative stress with male infertility
Authors: Mavrogianni D , Voitse A , Evgeni L , Stavros S , Drakakis P
Published: August 2021

Study question

Is GSTM1 polymorphism a putative biomarker of male infertility. Is there a possible correlation between GSTM1 presence, oxidative stress and male infertility?

Summary answer

A possible correlation may be established between GSTM1 polymorphism, and sperm mobility and morphology. Additionally oxidative potential may also be associated with fertility.

What is known already

Approximately 7% of men worldwide are affected by male infertility, which contributes to 40–50% of all infertility cases. Basic sperm analysis remains the main procedure of diagnosing male infertility, although there is still doubt whether it provides accurate clinical outcomes More accurate tests are essential for the diagnosis of male infertility. Oxidative stress is involved in the etiology of male infertility, with 30% to 80% of infertile men having increased levels of sperm free radicals. Recent research has shown that oxidative stress when combined with GSTM1-null genotype negatively affected the sperm quality of infertility group compared to the control group.

Study design, size, duration

Ninety semen samples were collected and divided into 2 groups. The study group consisted of sperm samples from 51 men identified as infertile according to WHO guidelines(case group).Sperm samples from 39 men with normal sperm count parameters (control group) were used for the control group.

Participants/materials, setting, methods

For all samples a sperm diagram was performed. and DNA was extracted. Polymerase chain reaction with specific for GSTM1 gene primers followed by agarose electrophoresis was applied to detect the presence of polymorphism. The MiOXSYS method was used to detect the oxidative potential.

Main results and the role of chance

This study shows that in the control group the presence of polymorphism was associated with a reduced number of immobile sperm cells (p = 0,035) while it appears to affect the normal morphology of the sperm(p = 0,042). In the infertility group the presence of the gene was significantly correlated with age(p = 0.046). No statistically significant difference was observed for the presence of the polymorphism between the 2 groups.In addition, we investigated the effect of oxidative potential with the MiOXSYS system and its relationship with sperm parameters. It was found that the two groups differed significantly when measuring oxidative potential, and that oxidoreduction potential affects sperm concentration/ml, total sperm count, type B motility and viscosity in the infertile male group.

Limitations, reasons for caution

A larger sample size could increase the accuracy of the results.

Wider implications of the findings: Studying the relation of the oxidative stress with sperm parameters may lead to the establishment of a genetic profile of increased risk of infertility, which would be of major importance especially in cases of idiopathic infertility.

Trial registration number

Not applicable
P–134 Zygote morphokinetic parameters (ZMP) differs between fertilized and non-fertilized (in vitro maturation) oocytes
Authors: Zhao M , Li H , Wang S , Alqawasmeh O , Xu M , Chung J , Chan D Y L
Published: August 2021

Study question

Is there any difference on the ZMP between the fertilized and the non-fertilized oocytes in an IVM cohort?

Summary answer

The zona pellucida (zp_g) , cytoplasm greyscale（cm_g),the cytoplasm size (cm_size), radius (cm_r) and deviation (cm_d) showed different patterns from the two groups.

What is known already

We developed a convolutional neural network (CNN) based algorithm that provides instant and bias-free analytical outcomes of oocyte morphological segmentation. The mature but not-fertilized oocytes tend to be static while the fertilized oocytes are more dynamic for preparing its following biological events.

Study design, size, duration

This was a case-control study on oocytes including 631 normal fertilized oocytes and 100 IVM oocytes from 01/08/2017 to 31/12/2019 conducted in the Prince of Wales Hospital, The Chinese University of Hong Kong.

Participants/materials, setting, methods

We used the convolutional neural network (CNN) algorithm to segment the ZMPs of the cytoplasm and zona pellucida of the oocytes. The ZMPs include cm_g, cm_size, cm_r, cm_d, zp_g, thickness of zona pellucida and the area of perivitelline space. For the ZMPs that did not change with time, we used t-test to test the significance and for the parameters changed with time we used dynamic warp timing and similarity test to find the difference.

Main results and the role of chance

The IVM group had a higher intensity of zp_g of 142.03 (128.52–158.70) compared with the fertilized group of 137.04 (121.69–154.37). The cm_g of IVM group was higher than fertilized group [122.55 (114.87–137.62) vs 119.37(108.88–132.87)]. The cm_size, cm_r and cm_d as parameters changed with time and showed a different pattern in two groups. The IVM group decreased the cm_size faster than the fertilized group but the fertilized group had a more dynamic change in the shape of cytoplasm (cm_d) during the development. The cm_r changed with the same pattern of cm_size provided evidence supporting the finding above.

Limitations, reasons for caution

The ZMPs in the IVM group was captured for 21 hours from the first polar body extrusion. Though the duration was similar to the one from fertilized to the first cleavage. The morphology change during that period may not represent the holistic one of IVM oocytes.

Wider implications of the findings: The IVM oocytes have different morphokinetic performance from fertilized oocytes. We used a novel method based on CNN to confirm the differences between the two groups showing that our algorithm was able to describe the morphokinetic changes in a quantitative way and corresponded with embryologist’s experience.

Trial registration number

The Hong Kong Obstetrical & Gynaecological Trust Fund
P–139 The role of hormones and LH receptor expression of granulosa cells collected from large and small follicles in natural/minimal stimulation cycle IVF
Authors: Okubo T , Teruaki H , Noriyuki O , Kenji O , Tomoya S
Published: August 2021

Study question

Do different follicle sizes influence gonadotropins (LH, FSH) and sex steroid (estradiol) in follicular fluids and LH receptor expression (LHCGR) in cumulus oocyte complexes (COCs)?

Summary answer

It was found that differences in levels of FSH, estradiol values and LHCGR mRNA expression level in COCs between small and large follicles.

What is known already

The maturity rate in oocytes of small follicle is significantly lower compared to that of large follicles.

Study design, size, duration

After obtaining written consents from 78 infertile patients, we aspirated the large (>15 mm) and small (<5 mm) follicles, and collected follicular fluids at oocyte retrieval.

Participants/materials, setting, methods

We measured levels of LH, FSH and estradiol by enzyme immunoassay from large and small follicular fluids after oocytes retrievals. All collected oocytes were distinguished from large and small follicles, we confirmed the maturity of retrieved oocytes by the presence of first polar body. Then we extracted total RNA from granulosa cells and measured mRNA expression of LHCGR, encoding the human LH receptor, by quantitative real-time PCR. Each value was normalized to ACTB mRNA levels.

Main results and the role of chance

LH levels were nearly equal between small and large follicles (P = 0.8356). Whereas FSH and estradiol levels were significantly lower in small follicles (P < 0.0001). The expression levels of LHCGR mRNA were significantly lower in small follicles than in large follicles during natural cycles. The maturity rate in oocytes of small follicle was significantly lower compared to that of large follicles (96.0% vs. 21.7%, P < 0001).

Limitations, reasons for caution

The main limitation of the present study was collected by 42 natural cycles and 36 mild stimulation cycles with letrozole following low-dose clomiphene.

Wider implications of the findings: In spite of almost the same LH levels between two groups, the reason why the significantly lower maturation rates of oocytes collected from small follicles is poor LHCGR mRNA expression due to insufficient granulosa cells glowth because of low FSH and estradiol levels.

Trial registration number

Not applicable
P–148 Effect of additional laser assisted drilling (LAD) during trophectoderm (TE) biopsy on mosaicism rate
Authors: Tee Z Q , Chan C W , Lim A Y X , Lee C S S
Published: August 2021

Study question

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

Summary answer

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

What is known already

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

Study design, size, duration

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

Participants/materials, setting, methods

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

Main results and the role of chance

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

Limitations, reasons for caution

Additional LAD during TE biopsy maybe be a confounding variable which affects the mosaicism rate. Moreover, the increase in mosaicism rate could be due to other unknown factors. A larger sample size is needed to confirm the results.

Wider implications of the findings: Based on our study, additional LAD during TE biopsy is not recommended as this may increase mosaicism rate. Biopsy should be done when the blastocyst has more herniated cell or when the ICM leaves the hatching point/biopsy site.

Trial registration number

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

Study question

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

Summary answer

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

What is known already

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

Study design, size, duration

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

Participants/materials, setting, methods

Part1:We compared the outputs of the automatic pronuclear detection system with these embryologists(three junior embryologists (1a), three intermediate embryologists (1b),and three senior embryologists (1c)) who had judged the pronuclei number from TL videos from 40 embryos each having 0,1,2,and 3PN.

Part2:The automatic pronuclear detection system determined the pronuclei number from the TL videos of 955 embryos scored as either 1,2,and 3PN,(different from those used in Part1),and the detection rate for 2PN was calculated.

Main results and the role of chance

Part1: The sensitivities for embryologist groups 1a),1b),1c) and the automatic pronuclear detection system were 80.0%,100%,100%,100% for 2PN, 60.0%,83.3%,86.7%,100% for 0PN, 46.7%,80.0%,86.7%,10.0% for 1PN, and 73.3%,96.7%,96.7%,10.0% for 3PN.

Part2: The precision for 2PN by the automatic pronuclear detection system was 99%.

Limitations, reasons for caution

In order to further improve the performance of the automatic pronuclear detection system, further adjustment of the algorithm and more training images will be utilised.

Wider implications of the findings: The detection of 2PN by the automatic pronuclear detection system was highly reliable, and the performance of the system was comparable to that of embryologists. These first results are reassuring and support the clinical use of the system as a further aid for embryologists, in routine laboratory practice.

Trial registration number

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

Study question

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

Summary answer

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

What is known already

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

Study design, size, duration

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

Participants/materials, setting, methods

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

Main results and the role of chance

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

Limitations, reasons for caution

Small number of samples

Wider implications of the findings: Further studies are needed to validate these findings for the diagnostic of sex embryos

Trial registration number

N/A