Sperm DNA Quality Research Articles

Genetic Variations in the NRF2 Microsatellite Contribute to the Regulation of Bovine Sperm-Borne Antioxidant Capacity.

Nuclear factor (erythroid-derived 2)-like 2 (NRF2) is a transcription factor protein-coding gene, considered a master regulator of the cellular stress response. The genetic variations of the NRF2 could influence its transcriptional profile and, subsequently, the stress resilience in all cell types, including sperm cells. Therefore, the sperm-borne antioxidants abundance in association with the genetic variation of a GCC microsatellite located at the 5' upstream region of the NRF2 gene was investigated in young (n = 8) and old (n = 8) Holstein bulls' sperm cells at different seasons. The sperm DNA was sequenced using Sanger sequencing, while- the sperm-borne mRNA analysis was carried out using the synthesized cDNA and qPCR. The data were statistically analyzed using GraphPad Prism 10.0.2 software. The results showed that two bulls had a heterozygous genotype of eight and nine GCC repeats, while biallelic of eight, nine, and fifteen repeats were identified in two, ten, and two bulls, respectively. The computational in silico analysis revealed that the NRF2 upstream sequence with 15, 9, and 8 GCC repeats bound with 725, 709, and 707 DNA-binding transcription factor proteins, respectively. Lower quality of sperm DNA was detected in the spring season compared to other seasons and in young bulls compared to old ones, particularly in the summer and autumn seasons. The mRNA expression analysis revealed that the PRDX1 gene was the abundant transcript among the studied sperm-borne antioxidants and was significantly determined in old bulls' spermatozoa. Moreover, two transcripts of the NRF2 gene and antioxidant (SOD1, CAT, GPX1, TXN1, NQO1) genes displayed differential expression patterns between the age groups across seasons in an antioxidant-dependent manner. The bulls with a heterozygous GCC sequence exhibited elevated sperm-borne mRNA levels of NRF2 and PRDX1 transcripts. Taken together, the findings suggest that the NRF2-GCC microsatellite may contribute to the transcription regulation of NRF2 transcripts and their subsequent downstream antioxidants in bovine sperm cells.

Role of DNase Activity in Human Sperm DNA Fragmentation.

In this clinical era of intracytoplasmic sperm injection (ICSI), where a single spermatozoon is chosen for fertilization, the diagnostic functionality of the classical parameters typically associated with fertilization, such as sperm concentration, sperm motility, acrosome integrity, and mitochondria, is perhaps becoming less critical. In contrast, the contribution of sperm DNA quality to our understanding of the impact of male fertility within the context of ICSI is gaining increasing interest and importance. Even with respect to natural conception, high levels of sperm DNA fragmentation (SDF) in the ejaculate can adversely affect reproductive outcomes. However, the precise origin of SDF pathology in sperm cells is often ambiguous and most likely to be multifactorial. Hence, the genetic makeup of an individual, unbalanced REDOX processes, enzymatic activity, environmental and lifestyle factors, and even damage during sperm handling in the laboratory all operate in a unique and often synergistic manner to produce or induce sperm DNA damage. Surprisingly, the contribution of active enzymes as potential agents of SDF has received much less attention and, therefore, is likely to be underrated. This review highlights the roles of different enzymes related to the degradation of sperm DNA as possible effectors of DNA molecules in spermatozoa.

Human circBOULE RNAs as potential biomarkers for sperm quality and male infertility.

Reliable molecular biomarkers to predict fertility remain scarce. The current study explored the potential of testis-specific circBOULE RNAs as biomarkers for male infertility and sperm quality. Using RT-PCR and RT-qPCR assays, we identified seven circular RNAs from the human BOULE gene in human sperm. We found that sperm circEx3-6 RNA exhibited a significantly decreased expression in asthenozoospermia while circEx2-6 and circEx2-7 expression decreased in teratozoospermia, compared with the controls. Furthermore, circEx2-6 expression exhibited a negative correlation with sperm DNA Fragmentation Index (DFI), and circEx2-7 levels were correlated with both fertilization and cleavage rates involving assisted reproductive technologies. Further functional analyses in a transgenic fly model lent support for the roles of circBOULE RNAs in sperm development and human fertility. Collectively, our findings support that sperm circBOULE RNAs may serve as diagnostic biomarkers for assessing sperm motility and DNA quality. Hence clinical application and significance of sperm circular RNAs in assisted reproductive technologies warrant further investigation.

#302 : Shorter Centrifugation Time Leads to Better Sperm Quality in Swim-Up Processing Technique

Background and Aims: Centrifugation is one of the many factors that cause sperm DNA damage in IVF semen preparation. This can occur due to the formation of reactive oxygen species (ROS) that are uncontrollable, so that the quality of sperm DNA can be damaged. This study aims to compare sperm quality results with swim-up technique under centrifugation times of 5 and 10 minutes. Method: This study involved 50 patients at the Halim Fertility Center clinic from June 2020 to October 2020. The samples obtained were processed using the swim-up method. Samples were divided into 3 groups, namely control, 5 and 10 minutes of centrifugation. Sperm quality recorded was concentration, total motility, progressive motility and DNA fragmentation index (DFI). Results: The sperm concentration after 5- and 10-min centrifugation before swim-up (27.78-39.79 and 35.36–51.09, respectively; p [Formula: see text] 0.05) was significantly higher compared to control (24.85–32.33). The total motility before and after 5- and 10-min centrifugation were 43.78–51.08, 97.66–98.20, and 97.86–98.20, respectively. The progressive motility after 5- and 10-min centrifugation (0–41 and 0–54, respectively) was significantly higher than control (0–24; p < 0.05). The DFI was significantly better after 5 min centrifugation (3.82–6.98) compared to control and after 10 min centrifugation (13.48–19.04 and 1–25, respectively; p < 0.05). Conclusion: Shorter centrifugation times indicate lower progressive motility than long centrifugation times, but considering the lower DFI level, shorter centrifugation times must be used in sperm preparation for IVF to produce better sperm quality.

The GGC Medium Reduces the DNA Fragmentation of Human Spermatozoa via in vitro Activation.

In vitro sperm stimulation by the addition of certain chemicals has become one of the most significant techniques for dealing with sperm DNA fragmentation, which is one of the leading causes of male infertility. The GGC medium is an invented triple antioxidant-containing medium (10 mM/ml green tea extract, 10 mM/ml glutathione, 60 mM/ml vitamin C, sodium pyruvate 0.01g/L, and 10% human serum albumin applied to 1L Ringer solution) for in vitro human sperm activation. This study aimed to evaluate the quality of the human sperm DNA after in vitro activation using a GGC medium. For this purpose, 200 semen samples were employed in this study. Before activation by swim-up technique, the samples were split into three groups, consisting of a control group (G1) with no activation medium, and G2 and G3 groups activated with Ferticult flushing medium and GGC medium, respectively. Afterward, the sperm DNA fragmentation index (DFI) was assessed pre- and post-swim-up activation. Based on the comparison of pre- and post-activation stages, the findings of DNA fragmentation revealed a significant increase at the pre-activation stage. Furthermore, relative to the other treatment groups, there was a strong significant (P<0.05) reduction of DFI in samples activated with GGC medium. The G2 and G3 showed a substantial reduction of DFI at the post-activation stage, compared to the pre-activation stage (P<0.05). According to the findings, both mediums can reduce DNA fragmentation; however, the GGC medium showed the most significant outcomes, compared to the Ferticult medium used for the in vitro activation of spermatozoa.

MP43-14 THE IMPACT OF VARICOCELE EMBOLISATION ON SPERM DNA FRAGMENTATION &amp; ASSISTED REPRODUCTIVE TECHNOLOGY (ART) OUTCOMES

You have accessJournal of UrologyCME1 Apr 2023MP43-14 THE IMPACT OF VARICOCELE EMBOLISATION ON SPERM DNA FRAGMENTATION & ASSISTED REPRODUCTIVE TECHNOLOGY (ART) OUTCOMES Adwoa Nyantakyi, Sheena Lewis, Yau Thum, Timothy Bracewell-Milnes, James Nicopoullos, Mohamed Al-Hussini, Jonathan Ramsay, Suks Minhas, and Tet Yap Adwoa NyantakyiAdwoa Nyantakyi More articles by this author , Sheena LewisSheena Lewis More articles by this author , Yau ThumYau Thum More articles by this author , Timothy Bracewell-MilnesTimothy Bracewell-Milnes More articles by this author , James NicopoullosJames Nicopoullos More articles by this author , Mohamed Al-HussiniMohamed Al-Hussini More articles by this author , Jonathan RamsayJonathan Ramsay More articles by this author , Suks MinhasSuks Minhas More articles by this author , and Tet YapTet Yap More articles by this author View All Author Informationhttps://doi.org/10.1097/JU.0000000000003289.14AboutPDF ToolsAdd to favoritesDownload CitationsTrack CitationsPermissionsReprints ShareFacebookLinked InTwitterEmail Abstract INTRODUCTION AND OBJECTIVE: A varicocele is a common and treatable cause of male infertility. Current guidelines advocate for treatment of varicoceles based on abnormal semenalysis rather than abnormal sperm DNA fragmentation (SDF). High SDF has been associated with poor pregnancy outcomes following ART. The aim of the study was to determine the outcomes of varicocele embolisation in an infertile male population on SDF and subsequent ART outcomes. METHODS: A retrospective study of 130 consecutive male partners of infertile couples with a clinical varicocele between 2015 and 2022 were included. 119 of these men underwent varicocele embolisation by a team of 2 radiologists, whilst 11 declined. SDF was assessed by the alkaline Comet test pre-intervention and at least 3 months after. 84 treated and the 11 untreated men proceeded to ICSI. Baseline couple parameters such as age, and pregnancy and live birth rate outcomes in the ART cycle following this intervention between groups were analysed. T & chi-square tests were used on continuous and categorical data respectively (significance rate p<0.01). RESULTS: Varicocele embolisation significantly reduced the proportion of sperm with DNA damage across all three Comet parameters (p<0.01) (Table 1). Only 18% of these men had normal semenalysis. Baseline parameters for men and women were similar with an average male age of 39 and female age of 37 for both groups. Couples who underwent embolisation had a significantly better pregnancy rate (65% vs 18%, p<0.01) and higher live birth rate which did not reach significance due to low numbers (32% vs 9%, p=0.11). CONCLUSIONS: There was a significant reduction in SDF following varicocele embolisation in this population of men of couples with infertility, with a significant difference in pregnancy rate outcomes in the treated group. Repair of clinical varicoceles improves sperm DNA quality, and can optimize fertility treatment success, even in patient with normal semenalysis. Limitations of the study include small numbers in the ‘untreated’ arm and lack of randomisation, a future avenue for research. Source of Funding: None © 2023 by American Urological Association Education and Research, Inc.FiguresReferencesRelatedDetails Volume 209Issue Supplement 4April 2023Page: e606 Advertisement Copyright & Permissions© 2023 by American Urological Association Education and Research, Inc.MetricsAuthor Information Adwoa Nyantakyi More articles by this author Sheena Lewis More articles by this author Yau Thum More articles by this author Timothy Bracewell-Milnes More articles by this author James Nicopoullos More articles by this author Mohamed Al-Hussini More articles by this author Jonathan Ramsay More articles by this author Suks Minhas More articles by this author Tet Yap More articles by this author Expand All Advertisement PDF downloadLoading ...

Decrease of air pollution during lockdown in Tuscany (Italy): An effect on sperm DNA fragmentation?

In March 2020, the Italian government imposed a national lockdown which was almost completely removed in June 2020. Due to the abrupt stop of human activities, emissions of air pollutants decreased. Air pollution is an environmental risk factor for noncommunicable disease and mortality. Emerging evidence also suggests a role in male infertility. In this study, we compared sperm DNA fragmentation (sDF) levels and conventional semen parameters between subjects undergoing sDF determination and routine semen analysis in a single Italian centre, during about 6 months before (N = 119) and after lockdown (N = 105). After lockdown, we found an improvement of sperm progressive motility (48.00[38.50-58.00]% vs. 42.00[33.00-53.00]%) and sDF levels (as total: 24.79[18.33-33.97]% vs. 35.02[25.04-45.73]%, p < .001; brighter: 14.02[10.69-17.93]% vs 18.54[13.58-25.82]%, p < .001 and dimmer sDF: 9.24[5.64-15.78]% vs. 12.24[8.08-19.10]%, p < .01), mirrored by a decrease of leukocyte semen concentration (p < .01). The improvement of sperm motility and DNA quality was maintained after adjusting for leukocyte concentration and several conditions known to affect sperm motility and/or sDF levels. With a significant decrease in air pollution observed in Tuscany during and after lockdown, associated improvement in sperm motility and DNA quality in patients referred to the infertility clinic is suggestive of the potential role of air pollution in male infertility.

High-throughput sperm DNA analysis at the single-cell and population levels.

Clinical semen quality assessment is critical to the treatment of infertility. Sperm DNA integrity testing provides critical information that can steer treatment and influence outcomes and offspring health. Flow cytometry is the gold standard approach to assess DNA integrity, but it is not commonly applied at the clinical level. The sperm chromatin dispersion (SCD) assay provides a simpler and cheaper alternative. However, SCD is low-throughput and non-quantitative - sperm assessment is serial, manual and suffers inter- and intra-observer variations. Here, an automated SCD analysis method is presented that enables quantitative sperm DNA quality assessment at the single-cell and population levels. Levering automated optical microscopy and a chromatin diffusion-based analysis, a sample of thousands of sperm that would otherwise require 5 hours is assessed in under 10 minutes - a clinically viable workflow. The sperm DNA diffusion coefficient (DDNA) measurement correlates (R2 = 0.96) with DNA fragmentation index (DFI) from the cytometry-based sperm chromatin structure assay (SCSA). The automated measurement of population-level sperm DNA fragmentation (% sDF) prevents inter-observer variations and shows a good agreement with the SCSA % DFI (R2 = 0.98). This automated approach standardizes and accelerates SCD-based sperm DNA analysis, enabling the clinical application of sperm DNA integrity assessment.

PROTECTIVE EFFECT OF L-PROLINE ON HUMAN SPERM CHARACTERISTICS DURING CRYOPRESERVATION

To explore the effectiveness of L-proline inclusion in freezing medium on sperm parameters, sperm DNA and chromatin quality, as well as the nitro-oxidative status of sperm culture medium in normozoospermic men who received antioxidants following the freezing-thawing process.

Sperm Selection With Hyaluronic Acid Improved Live Birth Outcomes Among Older Couples and Was Connected to Sperm DNA Quality, Potentially Affecting All Treatment Outcomes

(Abstracted from Hum Reprod 2022;37:1106–1125) With intracytoplasmic sperm injection (ICSI), many natural barriers that would normally prevent the entry of abnormal sperm are lost, increasing the importance of sperm DNA integrity for successful in vitro fertilization. There is currently no consensus on the optimal assay for measuring sperm DNA quality (DNAq), defined as any structural aspect of sperm chromatin that can compromise function if disrupted.

Revisiting the impact of varicocele and its treatments on male fertility

Varicocele is one of the most common, yet treatable, causes of male infertility. Varicoceles are present in more than 40% of infertile men with primary infertility, a figure that increases with age. Varicoceles impair semen parameters and sperm DNA and are linked with lower pregnancy and live birth rates. Until recently, men had seldom been examined in male fertility workups. This is changing as urologists have become recognized as team members in infertility. Hence identification and treatment are available as never before. Furthermore, as men become aware that they are as likely as their female partners to be infertile, they want equal ‘couple care’, requesting urological referrals as they realize that they can improve their semen quality and chances of fatherhood without or before fertility treatment. There is now a greater understanding of the mechanisms of varicocele-induced damage by oxidative stress, using sperm DNA as a sensitive biomarker of sperm quality. There is a current consensus that varicocele is linked to poor semen and repair improves semen and sperm DNA quality. Evidence is strengthening to indicate that varicocele repair increases pregnancy and live birth rates in natural conception and following fertility treatment.

P-099 The effect of sperm DNA quality on blastocyst and euploid rates

Abstract Study question Do sperm DNA quality measurements influence the blastocyst rate and/or euploid rates? Summary answer Sperm DNA fragmentation has a negative influence on the rate of blastulation. It does not impact on euploidy rates assessed by pre-implantation genetic testing (PGT-A). What is known already Some studies show that sperm DNA has no influence on embryo development and that its negative influence begins following implantation. Other studies show that every step of the fertility process from fertilization onwards is impacted by the male gamete as well as the female gamete. To date, data has been confounded by different sperm DNA tests and different clinical practices and parameters measured. Study design, size, duration This was a retrospective study using the sensitive Comet assay to analyse whether it’s three sperm DNA fragmentation scores influence embryo development or euploidy. Participants/materials, setting, methods Data from 110 cycles of intracytoplasmic sperm injection (ICSI) where the blastocyst rate (number of blastocysts/number of mature oocytes injected) was obtained from the patient database of a UK single centre. Inclusion criteria: Only outcomes from the first assisted reproductive cycle after a Comet assay had been performed were included. Included cycles were within one year of the test. Main results and the role of chance Blastocysts were ranked in 4 groups- 0.25-0.49, 0.5-0.74, 0.75-1.00. Three Comet scores were analysed: Average Comet Score (damage across the semen sample; ACS), Low Comet Score (proportion of sperm with good DNA; LCS) and High Comet Score ( proportion of sperm with high DNA damage; HCS). Clinical thresholds were calculated by comparison with fertile donor sperm. High ACS and LCS and low HCS were all associated with higher blastocyst rates (ANOVA, p = 0.013; Kruskal Wallis, p = 0.027, Post Hoc pairwise comparisons between blastocyst groups (p = 0.016-0.024). No relationship was found between euploid rate (number of euploid embryos/total embryo number) and sperm DNA damage p = 0.761). Limitations, reasons for caution Retrospective design requiring more data to clarify these associations coupled with the lack of randomisation and low power analysis. Wider implications of the findings This study reports the negative impact of sperm DNA fragmentation on blastocyst formation rates. Male partner diagnostic assessments such as sperm DNA quality testing are useful adjuncts to semen analysis. Further research to determine if there is a relationship between sperm DNA damage and euploid embryo outcomes is ongoing. Trial registration number IRB-001C06-01-22

P-083 The incidence of sperm DNA damage in men with chronic reproductive health issues

Abstract Study question What is the incidence of sperm DNA damage in men attending tertiary (urological) healthcare with long histories of infertility? Summary answer The majority of these men (95%), all with normal semen analyses had high levels of sperm DNA damage. What is known already Conventional semen analysis assesses sperm concentration, motility, and morphology but with a high degree of biological variability. Around 25% of men are given a diagnosis of unexplained infertility by normal semen analysis. The World Health Organisation (WHO) stated in 2021 that ‘clinically there is a growing awareness that chromosomal anomalies and gene mutations underlie a diverse spectrum of male infertility,’ so now recommends that sperm DNA is determined as an extended analysis. Sperm DNA damage occurs more often in infertile men and DNA damage is associated with recurrent pregnancy loss and decreased live birth rates following fertility treatment. Study design, size, duration A retrospective study of men attending a tertiary centre with productive failure had been offered a standard semen analysis and sperm DNA fragmentation using SpermComet technology. Participants/materials, setting, methods Results from 148 men tested between April 2017 and September 2021 were recruited from a tertiary urological database in the UK. Alongside a semen analysis, they also had a sperm DNA fragmentation test. Sperm DNA quality for the participants was assessed and reported as average, low and high DNA damage as a percentage compared with clinical thresholds used by Examen from their sperm data bank of 74 fertile sperm donors with recently achieved clinical pregnancies. Main results and the role of chance The average comet score was 37.8% +/-0.5, low comet score was 35.3% +/- 1.5 and high comet score was 20.2% +/-1.2. These values were all significantly outside ( p &amp;lt; 0.001, Mann Whitney test)) fertile parameters. 95% of these men presenting for infertility investigations had a significantly raised DNA damage across their semen sample with low proportions of healthy sperm and high levels of sperm with genomic damage. Limitations, reasons for caution Retrospective design, small sample size, lack of randomisation and low power analysis. Wider implications of the findings Sperm DNA damage was found in most of these men who, over years of investigation hadn't been offered tests other than semen analysis or therapy to improve their sperm quality. Thus, this tool could be a useful adjunct to semen analysis to guide the men to reproductive health. Trial registration number not applicable

P-033 A third of men with normospermia attending for initial fertility assessment could have DNA damage

Abstract Study question What is the incidence of sperm DNA damage in men with normal semen analyses attending for initial fertility investigation? Summary answer Half of men (54%) attending for infertilty investigation had a normal semen analysis of which a third (32%) of these had abnormal sperm DNA. What is known already Conventional semen analysis assesses sperm concentration, motility, and morphology but with a high degree of biological variability. Around 25% of men are given a diagnosis of unexplained infertility by normal semen analysis. The World Health Organisation (WHO) stated in 2021 that ‘clinically there is a growing awareness that chromosomal anomalies and gene mutations underlie a diverse spectrum of male infertility,’ so now recommends that sperm DNA is determined as an extended analysis. Sperm DNA damage occurs more often in infertile men and DNA damage is associated with recurrent pregnancy loss and decreased live birth rates following fertility treatment. Study design, size, duration A prospective cohort study of men attending a secondary fertility clinic were offered a standard semen analysis plus sperm DNA fragmentation using SpermComet technology. UK IRAS ethical approval was obtained. Men attending were given a patient information leaflet at their first appointment. The study duration was 6 months. In total, 142 men were recruited out of a possible maximum of 409 attendees. Participants/materials, setting, methods Participants included men who had been referred as a couple to a secondary level infertility clinic for an initial assessment. Information leaflets were either posted to them prior to their first appointment or given to them at the initial nurse-led consultation. Those who subsequently contacted our research team re-attended for a sperm analysis and an aliquot was cryopreserved and sent in weekly batches to Examen (Northern Ireland) for SpermComet DNA fragmentation assessment. Main results and the role of chance Between May and November 2021, there were 409 attendees of whom 142 consented to the study. Sperm DNA quality for the participants was assessed and reported as average, low and high DNA damage as a percentage compared with clinical thresholds used by Examen from their sperm bank of 63 fertile sperm donors who had recently achieved clinical pregnancies. Of the 142 recruited, 77 men (54%) attending for initial investigations had normospermia assessed against the WHO criteria, so were initially classified as unexplained. Of these, 25 (32%) had abnormal sperm DNA values. The average comet score was 39.7% +/-1.3, low comet score was 37.8% +/- 4.4 and high comet score was 17.0% +/-2.0. These values were all significantly outside ( p &amp;lt; 0.001) of the fertile parameters from 63 fertile donors used by Examen (fertile range: average &amp;lt;26%%, low &amp;gt;74% and high &amp;lt;4%). Our results suggest that up to a third of men reported to have normospermia will have DNA damaged sperm. This is important to consider before labelling a couple as 'unexplained'. Knowing the male's sperm DNA fragmentation assessment would allow for further discussion and exploration of lifestyle and dietary advice. Limitations, reasons for caution During the study period, not all attendees agreed to participate as it required an additional hospital visit. In this single site study, men have not yet been followed up for future fertility outcome such as natural conception and need or success of assisted conception. Wider implications of the findings Sperm DNA damage was found in a third of men who would previously have been classified as fertile and offered no further investigations or treatment to improve their sperm quality. Thus, this tool could be a useful adjunct to semen analysis to guide these couples’ future treatment pathways. Trial registration number not applicable

Relationship between sperm quality and total fertilization failure in intracytoplasmic sperm injection and in vitro fertilization cycles: A cross-sectional study.

BackgroundTotal fertilization failure (TFF) is associated with essential mechanistic and cellular events.ObjectiveThe present study is a comprehensive examination of detrimental effects with well-known assays for predicting TFF in conventional in vitro fertilization (IVF) and intracytoplasmic sperm injection (ICSI) cycles.Materials and MethodsSemen parameters of 90 men, including 60 cases who had experienced IVF/ICSI failure and a control group of 30 individuals, were evaluated. Sperm chromatin/DNA quality assessments were done by aniline blue, toluidine blue, chromomycin A3, and terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL) assays. A lipid hydroperoxide (LPO) kit was used to measure the LPO, and JC1 staining was used to evaluate mitochondrial membrane potential (MMP).ResultsThere were statistically significant differences found between the IVF, ICSI and control groups by the toluidine blue (p = 0.01), TUNEL (p = 0.02), and chromomycin A3 (p 0.001) tests, but not by the aniline blue staining. Furthermore, there was a significant difference regarding LPO concentration and high MMP in cases of IVF fertilization failure compared to the control group (p = 0.04, p = 0.02, respectively). The logistic regression model showed that sperm viability was predictive for fertilization failure in the ICSI group. Sperm chromatin and DNA quality assays were not predictors for TFF in either group.ConclusionCellular events such as high DNA fragmentation damage, high levels of reactive oxygen species, and low MMP levels can cause TFF in IVF and ICSI programs. Diagnostic tests, especially in cases with previous fertilization failure, showed significant differences in sperm chromatin and DNA quality between groups but could not predict the risk of TFF.

Impact of heavy alcohol consumption and cigarette smoking on sperm DNAintegrity.

The purposes of the presents study were to investigate the impact of alcohol consumption and cigarette smoking on semen parameters and sperm DNA quality, as well as to determine whether tobacco smoking, or alcohol consumption causes more deterioration of sperm quality. Two hundred and eleven semen samples of men were included in this study. Four groups were studied: heavy smokers (N=48), heavy drinkers (N=52), non-smokers (n=70), and non-drinkers (n=41). Semen parameters were determined according to WHO guidelines, protamine deficiency assessed by chromomycin (CMA3) staining, and sperm DNA fragmentation (sDF) evaluated by TUNEL assay. Sperm parameters were significantly higher in non-smokers versus smokers and in non-drinkers versus drinkers (p < 0.005). However, protamine deficiency and sDF were significantly lower in non-smokers versus smokers and in non-drinkers versus drinkers (p < 0.0001). No significant difference in the semen analysis parameters was observed between heavy smokers and heavy drinkers (semen volume: 3.20 ± 1.43 vs. 2.81 ± 1.56 ml, semen count: 65.75 ± 31.32 vs. 53.51 ± 32.67 mill/ml, total motility: 24.27 ± 8.18 vs. 23.75 ± 1.75%, sperm vitality: 36.15 ± 18.57 vs. 34.62 ± 16.65%, functional integrity: 41.56 ± 18.57 vs. 45.96 ± 17.98% and the morphologically normal spermatozoa: 28.77 ± 11.82 vs. 27.06 ± 13.13%, respectively). However, protamine deficiency was significantly higher among drinkers than smokers (37.03 ± 9.75 vs. 33.27 ± 8.56%, p=0.020). The sDF was also significantly higher among drinkers than smokers (22.37 ± 7.60 vs. 15.55 ± 3.33%, p < 0.0001). Thus, cigarette smoking, and heavy alcohol intake can deteriorate sperm quality. However, alcohol consumption deteriorates sperm maturity and damages DNA integrity at significantly higher rates than cigarette smoking.

Sperm selection with hyaluronic acid improved live birth outcomes among older couples and was connected to sperm DNA quality, potentially affecting all treatment outcomes.

Sperm selection with hyaluronic acid improved live birth outcomes among older couples and was connected to sperm DNA quality, potentially affecting all treatment outcomes.

Investigation of pathology associated with Chlamydia pecorum infection in the male reproductive tract, and the effect on spermatogenesis and semen quality in the koala (Phascolarctos cinereus)

There is growing evidence that Chlamydia pecorum infection of the male koala reproductive tract causes inflammation and pathology of the urogenital tract. Previous studies have revealed that male koalas exhibiting severe clinical signs of urogenital chlamydial disease had an increased incidence of sperm DNA fragmentation and abnormal sperm morphology, suggestive of chronic exposure to C. pecorum infection and/or inflammation in the testis and epididymis, with residual pathology and lesions disrupting spermatogenesis and maturation of spermatozoa. This study specifically aimed to determine whether pathology associated with chlamydial infection in different regions of the male koala reproductive tract had an adverse effect on classical seminal parameters, sperm DNA quality and endocrine function (testosterone secretion) of naturally infected males. Semen from 58 sexually mature male koalas deemed not suitable for rehabilitation or treatment was assessed, in addition to undertaking a GnRH challenge to determine the androgenic capacity of the testis. Following euthanasia, tissue samples from testes, epididymis and prostate were evaluated for histopathology and real time polymerase chain reaction (qPCR). A significant difference in sperm concentration was observed between males with unilateral and bilateral testicular atrophy and C. pecorum infection (P = 0.011); and between males with unilateral atrophy and C. pecorum infection in one testis and bilateral normal testes with no C. pecorum infection (P = 0.008). No significant association was found for any other semen parameters when categorised by histopathology and C. pecorum tissue presence within the testes, epididymis and prostate. Plasma testosterone concentrations did not significantly differ between testicular histopathology diagnosis and/or C. pecorum infection status. This study suggests Chlamydia infection and inflammation may not be the predominant reason of disruption to spermatogenesis in the wild koala but rather testicular degeneration and atrophy, irrespective of Chlamydia infection, appears to be the primary reason of decreased sperm concentration.

Beneficial effects of hypotaurine supplementation in preparation and freezing media on human sperm cryo-capacitation and DNA quality

BackgroundAlthough widely used, slow freezing considerably modifies the functions of human spermatozoa. Cryopreservation induces nuclear sperm alterations and cryo-capacitation, reducing the chances of pregnancy. Hypotaurine is naturally present in the male and female genital tracts and has capacitating, osmolytic and anti-oxidant properties. The analysis were performed on surplus semen of men with normal (n = 19) or abnormal (n = 14) sperm parameters. Spermatozoa were selected by density gradient centrifugation before slow freezing. For each sample, these steps were performed in parallel with (“H+” arm) or without (“H-” arm) hypotaurine supplementation. After thawing, we measured total and progressive mobility, vitality, acrosome integrity, markers of capacitation signaling pathway and nuclear quality. For the latter, we focused on sperm chromatin packaging, DNA fragmentation and the presence of vacuoles in the sperm nucleus.ResultsPost-thaw spermatozoa selected and frozen in the presence of hypotaurine had a higher vitality (+ 16.7%, p < 0.001), progressive and total motility (+ 39.9% and + 21.6% respectively, p < 0.005) than spermatozoa from the control “H-” arm. Hypotaurine also reduced the non-specific phosphorylation of the capacitation protein markers P110 and P80 (p < 0.01), indicating a decrease in cryo-capacitation. Hypotaurine supplementation reduced chromatin decondensation, measured by chromomycin A3 (− 16.1%, p < 0.05), DNA fragmentation (− 18.7%, p < 0.05) and nuclear vacuolization (− 20.8%, p < 0.05).ConclusionOur study is the first to demonstrate beneficial effects of hypotaurine supplementation in preparation and freezing procedures on human spermatozoa sperm fertilization capacity and nucleus quality. Hypotaurine supplementation limited cryo-capacitation, increased the proportion of live and progressively motile spermatozoa and reduces the percentage of spermatozoa showing chromatin decondensation, DNA fragmentation and nuclear vacuolation.Trial registrationClinical Trial, NCT04011813. Registered 19 May 2019 - Retrospectively registered.

Sperm DNA Fragmentation.

Sperm DNA fragmentation refers to the accumulation of adducts as well as single- or double-strand DNA breaks and reflects the sperm DNA quality. Current data suggest that there are differences in sperm DNA quality among individuals with high or low fertility, and this observation has led to the idea that testing sperm DNA fragmentation could be a useful test of male fertility. However, sperm DNA fragmentation has become one of the most frequently debated topics in reproductive medicine, as there is no agreement about the optimal way to test for DNA fragmentation, the clinically significant level of sperm DNA fragmentation, as well as the best therapeutic options for infertile men. This article presents current evidence related to sperm DNA fragmentation and its role in managing male infertility.