Semen Reactive Oxygen Species Levels Research Articles

P-041 Impact of cigarette smoking on level of sperm reactive oxygen species in normozoospermic men

Abstract Study question Does cigarette smoking influence the levels of sperm reactive oxygen species (ROS) in men with normal semen routine parameters? Summary answer Cigarettes impact the sperm ROS of normozoospermic smokers, whose sperm ROS level is elevated when they smoke equal to or more than 10 cigarettes daily. What is known already Increasing evidence indicates that oxidative stress (OS) plays an independent role in the etiology of male infertility, with 30% to 80% of infertile men having elevated ROS levels in semen. Cigarette smoking leads to an imbalance between ROS and antioxidants in the semen of smokers, which may negatively affect semen quality and male fertility. However, few studies concern whether cigarette smoking would influence the sperm ROS levels in normozoospermic men. Study design, size, duration Data of semen analysis from January 2021 to December 2023 were involved. Men with normal semen routine parameters, including sperm concentration, total sperm number, sperm progressive motility, total motility and normal forms of sperm morphology, were included in the study. Sperm ROS levels of normozoospermic men were retrospectively analyzed. Men with varicocele and leucocytospermia were excluded. Participants/materials, setting, methods Sperm count and motility were analyzed by the computer-assisted sperm analysis (CASA) system, while sperm morphology was examined after the Diff-Quik rapid staining procedure. Sperm ROS was measured by flow cytometry using the probe DCFH-DA. Sperm ROS levels of smokers and non-smokers were compared by the Mann-Whitney U test. The Kruskal-Wallis H test was used for the comparison of sperm ROS levels among the control group and smoking subgroups. Main results and the role of chance A total of 2946 subjects were involved in the present study. The included subjects were divided into smokers (n = 730) and non-smokers (n = 2216) according to the clinical records. And the included smokers were subsequently divided into three subgroups according to the daily cigarettes taken, which were group A (1∼9 cigarettes/day, n = 184), group B (1∼19 cigarettes/day, n = 315), and group C (>20 cigarettes/day, n = 231). No statistically significant differences in age or BMI were found between smokers and non-smokers with normozoospermia. The sperm ROS level of the smoker was higher than that of the non-smoker [14.53% (14.84%) vs. 13.07% (12.09%), p = 0.001]. Compared to the control group (non-smoker), the sperm ROS levels of group B [13.07% (12.09%) vs. 15.44% (14.28%), p = 0.002] and group C [13.07% (12.09%) vs. 14.80% (14.31%)., p = 0.042] were significantly elevated. ROS levels in the control group and group A were comparable [13.07% (12.09%) vs. 13.46% (15.74%)., p = 0.391]. Limitations, reasons for caution It is a single-center study and the above results were from a retrospective analysis. Wider implications of the findings The above results indicate that cigarette smoking, although it would not deteriorate semen routine parameters, may increase the ROS level in sperm. Elevated ROS in sperm may lead to an imbalance of redox in sperm, which may contribute to male infertility. Trial registration number not applicable

P-099 The development of a novel sperm washing medium, enriched with serotonin as the active constituent, with the objective of enhancing sperm motility and viability

Abstract Study question Is it possible to develop a novel product that increases the success rate of treating male infertility through assisted reproductive techniques? Summary answer Sperm washing media, aiming to improve sperm quality, enhance sperm motility, and generally support male reproductive health in the field of assisted reproductive techniques. What is known already Infertility impacts 15% of all couples globally, with 50% of infertility cases attributed solely to male factors. Reduced semen motility is identified as a primary factor directly associated with infertility. The utilization of supplementation to enhance overall sperm quality has gained increasing popularity worldwide. The sperm preparation process in Assisted Reproductive Technology (ART) is crucial for successful fertilization, with sperm washing mediums playing a critical role in enhancing efficiency. Sperm-washing media is a specialized solution utilized in assisted reproductive technologies such as intrauterine insemination and in vitro fertilization to remove the seminal fluid and debris for sperm to undergo capacitation. Study design, size, duration In this study, 200 normozoospermic men were included, and their semen samples were analyzed over an 18-month period following the guidelines outlined by the World Health Organization (2010) to assess the efficacy of serotonin, selenium, zinc, L-Carnitine, and Coenzyme Q10 supplementation on sperm motility, viability, ROS and DNA damage. Participants/materials, setting, methods Sperm samples obtained from 200 volunteer men undergoing infertility assessments were subdivided into three samples.The sperm samples underwent preparation using the swim-up technique. The study involved the examination of samples in three distinct washing media: (1) Origio, (2) Irvine Scientific, (3) a prototype (serotonin, selenium, zinc, L-carnitine, and Coenzyme Q10). Each medium was analyzed for motility, apoptosis, reactive oxygen species (ROS), and DNA damage. Main results and the role of chance It is found that mean basal total sperm counts are 38 million. After sperm washing, mean total sperm counts of Origio®, Irvine® and Prototype are 15 million respectively (p > 0.05). As total progressive motility is % 40 before washing, it is increased to 62%,56% and 64% after washing with Origio®, Irvine® and Prototype, respectively (p > 0.05). Annexin V staining is used for detection of apoptosis in sperm samples. On average, early apoptosis over the total sperm count(TSC) per subgroups are 34.67% for Prototype, 33.17% for Origio® and 30.96% for Irvine®, whereas late apoptosis ratios are found significantly lower in Prototype compared to Origio and Irvine; 8.7%, 28.26% and 27.74%, respectively. Ratio of necrosis over TSC per subgroups is 3.15% for Prototype, 13.49% for Origio® and 18.78% for Irvine® whereas viability ratios are as 53.48%, 25.08%, 22.52%, respectively.(p < 0.05). ROS levels in semen after washing are significantly lower for Prototype (5.9%, 10.29%, and 18.22%, Prototype, Origio® and Irvine®, respectively) (p < 0.05). Limitations, reasons for caution Semen samples from males aged 28 to 45, who are unable to conceive, do not smoke, and have not undergone any surgical interventions, are collected from patients after a sexual abstinence period of 3-4 days. Wider implications of the findings We propose that our serotonin-activated washing media for sperm preparation can serve as a suitable choice for selecting high-quality sperm in assisted reproductive technology programs. These molecules may enhance the success of assisted reproduction techniques when present in sperm preparation media. Trial registration number SerotoneX

PMON259 Weight Loss using a Formula Low Energy Diet Reduces Seminal Oxidative Stress in Men with Obesity and Oligospermia: a Randomised Controlled Study

Abstract Background Poor sperm quality (male infertility) is present in 50% of couples requiring assisted reproductive technologies (ART). Few couples worldwide have access to ART, and there are currently no available drugs to improve sperm function. Obesity is a major cause of male infertility. Semen reactive oxygen species (ROS), is a recently identified mechanism for sperm DNA damage resulting in reduced fertilisation and pregnancy outcomes. Semen levels of ROS are positively correlated with body mass index. However, the acute energy restriction following bariatric surgery may paradoxically reduce sperm function, making it an unsuitable treatment for male infertility. No previous randomised controlled study has been performed investigating the effects of weight loss on sperm function and semen ROS in men with obesity and infertility. We hypothesized that milder weight loss through energy restriction could be a novel treatment to reduce semen ROS and improve sperm function in men with obesity and oligospermia. Objective Determine effects of a formula low energy diet (LED) on semen parameters and ROS in men with obesity and oligospermia. Methods Thirty-six men with obesity and oligospermia participated in a 16-week randomised, controlled study. After 2-weeks of baseline observation, all men were randomised to either LED (∼800kcal/day) or a control intervention of generalised healthy eating advice lasting 16-weeks; WHO semen analysis, semen ROS, metabolic parameters and reproductive hormones were monitored throughout the study. Semen ROS was measured in relative light years/sec/million sperm (RLU/s/mill) using an in-house, validated chemiluminescent luminol-based assay. Results At baseline, men with obesity and oligospermia had significantly elevated semen ROS levels when compared with the reference range generated from healthy men with normal BMI (median (inter-quartile range) ROS in RLU/s/mill: <3.8, healthy men; 60.3(113.1), control at baseline; 82.1(100.6) LED). Participants on the LED lost 9-fold more weight compared with control intervention (weight ± SD in kg: -1.5±4.3, control; -14.3±4.5, LED, p<0.0001). After 16 weeks, median semen ROS was reduced 12-fold greater in the LED versus control group (change in ROS vs. baseline in RLU/s/mill: -0.2(61.9), control; -6.0(137.5), LED; p<0.05). Total and progressive motility, and total motile count increased in the LED group versus the control intervention, but these changes were non-significant. Conclusions This is the first randomised controlled study investigating effects of dietary weight loss on spermatogenesis in men with obesity and oligospermia. Our study provides proof-of-concept, and that weight loss could be used therapeutically in men with obesity and infertility, to reduce seminal ROS which is associated with increased sperm DNA damage. These data have important potential implications for management of couples with male infertility. Larger studies are needed to investigate whether weight loss is sufficient to improve pregnancy outcomes in couples affected by male infertility due to obesity. Presentation: Monday, June 13, 2022 12:30 p.m. - 2:30 p.m.

Exploring the Role of Oxidative Stress in Sperm Motility: A Proteomic Network Approach.

Significance: Infertility is a major global health problem, with nearly half of the cases being associated with male factors. Although reactive oxygen species (ROS) are crucial for sperm cell normal physiological processes, an imbalance between ROS production and antioxidants can lead to oxidative stress that can impair sperm function. Indeed, high semen ROS levels are reported in 30%-80% of infertile men. Recent Advances: Male oxidative stress infertility is an uprising classification for idiopathic infertility. Proteomic approaches, including quantitative mass spectrometry (MS)-based proteomics, are being utilized to explore the molecular mechanisms associated with oxidative stress in male infertility. Critical Issues: In this review, proteome data were collected from articles available on PubMed centered on MS-based proteomic studies, performed in seminal plasma and sperm cell samples, and enrolling men with impaired semen parameters. The bioinformatic analysis of proteome data with Cytoscape (ClueGO+CluePedia) and STRING tools allowed the identification of the biological processes more prevalent in asthenozoospermia, with focus on the ones related to oxidative stress. Future Directions: The identification of the antioxidant proteins in seminal plasma and sperm cells that can protect sperm cells from oxidative stress is crucial not only for a better understanding of the molecular mechanisms associated with male infertility but specially to guide new therapeutic possibilities. Antioxid. Redox Signal. 37, 501-520.

Chlamydia trachomatis Infection: its relation to semen parameters and sperm DNA integrity

The present study was conducted to determine the effect of Chlamydia trachomatis (CT) infection on semen quality and sperm DNA integrity. The study included 60 infertile male patients with CT infection and 25 age matched controls. Diagnosis of patients was based on detection of CT IgA by ELISA in seminal plasma and CT plasmid DNA in the semen sediment. All patients and controls were subjected to the following investigations: history taking, conventional semen analysis, detection of CT IgA, Plasmid DNA in semen samples, reactive oxygen species (ROS) and percentage of DNA fragmentation. There was significant increase in semen ROS levels and the percentage of sperm DNA fragmentation in the CT patient group when compared to the control group (P<0.05) and in those with leukocytospermia when compared to those without leukocytospermia (P<0.05). In the patient group with CT infection there was a positive correlation between the percentage of DNA fragmentation, ROS (r = 0.82 with P<0.0001) and pus cell count. (r = 0.7 with P<0.0001). In patients with leukocytospermia, there was a positive correlation between the percentage of DNA fragmentation, ROS (r = 0.9 with P<0.0001) and pus cell count (r = 0.83 with P<0.0001). In conclusion, sperm concentration, mobility, and viability, are significantly decreased in patients with CT compared to controls. ROS levels and the percentage of sperm DNA fragmentation significantly increased in CT patients especially patients with leukocytospermia.

Semen reactive oxygen species levels are correlated with erectile function among chronic prostatitis/chronic pelvic pain syndrome patients.

Chronic Prostatitis/Chronic Pelvic Pain Syndrome (CP/CPPS) is often associated with erectile dysfunction (ED). However, the underlying pathophysiological mechanisms of ED occurrence are still unclear in patients with CP/CPPS. The aim of the study was to investigate superoxide anion (O2•-) and total reactive oxygen species (ROS) production in semen of men with category IIIA CP/CPPS and their association with ED. This prospective study included 33 men with category IIIA CP/CPPS. Control group consisted of 13 healthy men. Total ROS and O2•- production were assayed by luminol and lucigenin-dependent chemiluminescence (CL) methods, respectively. ED was evaluated using the IIEF-5 questionnaire. Patients with CP/CPPS had significantly higher seminal total ROS and O2•- levels than healthy control subjects (2.9 ± 0.5 relative light unit (RLU) vs. 2.4 ± 0.2 RLU, p < 0.001; luminol-dependent CL and 2.5 ± 0.4 RLU vs. 2.3 ± 0.2 RLU, p = 0.02; lucigenin-dependent CL, respectively). Seminal O2•- and ROS levels were negatively correlated with IIEF-5 scores (r = -0.556, r = -0.536; p < 0.001, respectively). These results may suggest O2•-/ROS overproduction could be one of the important mechanisms in the etiology of ED development in CP/CPPS patients.

MP15-02 SEMEN REACTIVE OXYGEN SPECIES LEVELS IN PATIENTS WITH CHRONIC PROSTATITIS/CHRONIC PELVIC PAIN SYNDROME CATEGORY IIIA: RELATIONSHIP WITH THE NATIONAL INSTITUTES OF HEALTH-CHRONIC PROSTATITIS SYMPTOM INDEX

You have accessJournal of UrologyInfections/Inflammation/Cystic Disease of the Genitourinary Tract: Prostate & Genitalia I1 Apr 2018MP15-02 SEMEN REACTIVE OXYGEN SPECIES LEVELS IN PATIENTS WITH CHRONIC PROSTATITIS/CHRONIC PELVIC PAIN SYNDROME CATEGORY IIIA: RELATIONSHIP WITH THE NATIONAL INSTITUTES OF HEALTH-CHRONIC PROSTATITIS SYMPTOM INDEX Ilter Alkan, Meral Yüksel, Anil Atalay, Lütfi Canat, çigdem Arabaci, Muammer Bozkurt, and Murad Basar Ilter AlkanIlter Alkan More articles by this author , Meral YükselMeral Yüksel More articles by this author , Anil AtalayAnil Atalay More articles by this author , Lütfi CanatLütfi Canat More articles by this author , çigdem Arabaciçigdem Arabaci More articles by this author , Muammer BozkurtMuammer Bozkurt More articles by this author , and Murad BasarMurad Basar More articles by this author View All Author Informationhttps://doi.org/10.1016/j.juro.2018.02.515AboutPDF ToolsAdd to favoritesDownload CitationsTrack CitationsPermissionsReprints ShareFacebookTwitterLinked InEmail INTRODUCTION AND OBJECTIVES The etiology of CP/CPPS is still unclear despite many studies suggests the possible role of infectious, inflammatory, immunologic and neuronal causes. Reactive oxygen species (ROS) induced oxidative stress (OS) could be an etiologic factor in CP/CPPS. We would like to determine if there is any difference between seminal ROS levels in patients with CP/CPPS National Institutes of Health (NIH) category IIIA and age-matched healthy men and also determine any positive relationship between seminal ROS levels and disease symptom severity scored by The Turkish version of NIH-Chronic Prostatitis Symptom Index (NIH-CPSI). METHODS We prospectively evaluated 63 patients for prostatitis-like symptoms. Thirty-three patients (age, mean ± SD; 41.1 ± 9.75) provided study inclusion criteria for CP/CPPS category IIIA and enrolled into the study. Eight age-matched (age: 40.7 ± 1.98) healthy men consist control group. The Turkish version of NIH-CPSI form was completed by the study participants. Semen characteristics were examined according to the WHO 2010 criteria. No patients had leukocytospermia (≥ 1.0 x106 white blood cells/mL). Swim-up method was used for the sperm preparation. ROS levels were measured by chemiluminescence method via luminol enhancer. Results are given as relative light unit (rlu) /106 spermatozoa /mL. The log of (ROS+1) was used in calculations. RESULTS The mean sperm concentrations (106/mL) and motilities (%) of patient group and control subjects were 121 ± 116 x 106/mL, 35.9 ± 9.94 % vs. 110.8 ± 77.37 x 106/mL, 39.6 ± 7.19 %, respectively. No statistically significant differences were found in age, semen sperm concentrations and motility in between CP/CPPS patients and the control subjects. (p = 0.903, 0.755 and 0.2001, respectively) CP/CPPS patients had higher luminol enhanced ROS levels 2.9 ± 0.45 than healthy control subjects 2.5 ± 0.20 (Mann-Whitney test, p = 0.002). We found that there was a moderately positive correlation between ROS levels and NIH-CPSI scores.(Spearman Correlation Test, r = 0.291, p=0.048) CONCLUSIONS Semen ROS levels in CP/CPPS category IIIA patients were significantly higher than healthy control subjects. Overproduction of ROS or OS could be an etiologic factor in this group of patients. Semen ROS levels were positively correlated with NIH-CPSI scores, it could be another tool for assessing disease severity in CP/CPPS category IIIA patients. © 2018FiguresReferencesRelatedDetails Volume 199Issue 4SApril 2018Page: e189 Advertisement Copyright & Permissions© 2018MetricsAuthor Information Ilter Alkan More articles by this author Meral Yüksel More articles by this author Anil Atalay More articles by this author Lütfi Canat More articles by this author çigdem Arabaci More articles by this author Muammer Bozkurt More articles by this author Murad Basar More articles by this author Expand All Advertisement Advertisement PDF downloadLoading ...

New Approaches to Boar Semen Evaluation, Processing and Improvement.

The improvement of boar reproductive performance may be the next frontier in reproductive management of swine herd in Unites States, facilitated by better understanding of boar sperm function and by the introduction of new advanced instrumentation in the andrology field. Objective single ejaculate evaluation and individual boar fertility prediction may be possible by introducing automated flow cytometric semen analysis with vital stains (e.g. acrosomal integrity and mito-potential), DNA fragmentation analysis and biomarkers (ubiquitin, PAWP, ALOX15, aggresome) associated with normal or defective sperm phenotypes. Measurement of sperm-produced reactive oxygen species (ROS) is a helpful indicator of normal semen sample. Semen ROS levels could be managed by the addition of ROS-scavenging antioxidants. Alternative energy regeneration substrates and sperm stimulants such as inorganic pyrophosphate and caffeine could increase sperm lifespan in extended semen and within the female reproductive system. Such technology could be combined with timed sperm release in the female reproductive system after artificial insemination. Sperm phenotype analysis by the image-based flow cytometry will go hand in hand with the advancement of swine genomics, linking aberrant sperm phenotype to the fertility influencing gene polymorphisms. Finally, poor-quality ejaculates could be rescued and acceptable ejaculates improved by semen purification methods such as the nanoparticle-based semen purification and magnetic-activated sperm sorting. Altogether, these scientific and technological advances could benefit swine industry, provided that the challenges of new technology adoption, dissemination and cost reduction are met.

Evaluation of the impact of leucocytospermia on semen oxidative status by chemiluminescence technique in infertile men

The presence of high reactive oxygen species (ROS) levels in semen is a major factor involved in the decline of male fertility. In seminal plasma, ROS are mainly produced by activated leucocytes. Spermatozoa were the first cell type reported to show a potential susceptibility to oxidative damage. The aim of our study was to evaluate the impact of leucocytospermia on basal and FMLP (Formyl-Methionyl-Leucyl-Phenylalanine) induced oxidative status in semen of infertile men. We also analyzed the correlations of the spermatic parameters with amounts of ROS in semen. Our study included 50 semen samples of infertile men. Sperm analysis was performed using WHO standardized method. Seminal leucocytes were quantified using peroxidase technique. The measurement of ROS levels in semen was made by chemiluminescence assay. We measure respectively ROS amounts in neat semen and in washed sperm cells suspension from the same ejaculate. We also applied the test of provocation of leucocytes by FMLP on neat and washed samples to assess the spermatic oxidative status after leucocyte stimulation. Our results showed significant correlations between ROS levels in neat semen and many sperm parameters: motility, sperm concentration, leucocytes concentration and the rate of sperm cytoplasmic droplets. The studied samples were divided into 2 groups: (G1) composed of 36 samples without leucocytospermia and (G2) composed of 14 leucospermic samples. ROS levels were significantly lower in G1 than in G2 (p = 0.002). ROS production was significantly increased after application of FMLP in washed leucospermic samples (p = 0.001). The measurement of ROS in neat semen is a considerable contribution to explore the impairment of semen quality in infertile men. ROS levels in washed semen reflect the oxidative status generated by sperm preparation techniques used in assisted reproductive procedures. Levels of ROS are highly influenced by the presence of leucocytes and associated with decreased seminal parameters.

Effect of Smoking on Sperm Vitality, DNA Integrity, Seminal Oxidative Stress, Zinc in Fertile Men

To assess the effect of smoking on sperm vitality, sperm DNA integrity, semen reactive oxygen species, and zinc levels in fertile men. One-hundred sixty men were investigated. They were divided into 2 equal groups: healthy fertile nonsmokers and healthy fertile smokers. They were subjected to history taking, clinical examination, and semen analysis. In their semen, sperm hypo-osmotic swelling test, sperm DNA fragmentation test, seminal reactive oxygen species, and zinc were assessed. Compared with fertile nonsmokers, fertile smokers were significantly associated with lower hypo-osmotic swelling test and seminal zinc levels and significantly associated with higher sperm DNA fragmentation percent and seminal reactive oxygen species levels. Smoking (cigarettes/day and duration) has detrimental effects on sperm motility, viability, DNA fragmentation, seminal zinc levels, and semen reactive oxygen species levels, even in fertile men, and it is directly correlated with cigarette quantity and smoking duration.

Sperm DNA damage in patients with chronic viral C hepatitis

Introduction The aim of this study was to evaluate the conventional and biofunctional parameters of sperm in young infertile patients with Hepatitis C (HCV) infection. Methods Forty HCV patients with primary infertility, aged 27 to 42 years (mean 36.4 years) and twenty HCV patients with secondary infertility aged 28 to 45 years (mean 35.0 ± 2.8 years), underwent hormonal and sperm analysis in addition to the determination of reactive oxygen species (ROS) concentrations in the sperm and flow-cytometric evaluation. The following biofunctional sperm parameters were evaluated by flow cytometry: DNA fragmentation, mitochondrial membrane potential, chromatin condensation, and the rate of early apoptosis. Results Overall, patients with HCV showed significantly worse median values of conventional and biofunctional sperm parameters than control subjects, including sperm density (31.7 vs. 80.4 million/ml), forward motility (9.4 vs. 25%), normal forms (15.4 vs. 24.8%), DNA fragmentation (6.6 vs. 2.2%), low MMP (45.5 vs. 8%), an early apoptosis rate (5 vs. 2.7%), and abnormal chromatin (18.9 vs. 13.9%). Finally, HCV patients had significantly higher basal (250 vs. 75 × 10 3/cpm) and stimulated (550 vs. 120 × 10 3/cpm) ROS levels in semen compared to control subjects. None of the examined parameters (sperm, hormonal, biofunctional and assessment of oxidative status in the semen) was significantly different between HCV patients with primary and secondary infertilities. Discussion These results confirm that HCV infection has a negative impact on sperm parameters. The overlap of the results observed in the two groups of HCV patients supports the hypothesis that HCV infection may cause to alterations in sperm parameters.

Sperm DNA damage vs. reactive oxygen species (ROS) – is there evidence of a causal relationship?

Assessments of sperm DNA damage are thought to measure DNA decondensation that is in large part due to peroxidative damage. ROS levels are checked often; studies have shown that treatment of patients with high ROS levels in semen can improve overall semen parameters. However, there exists little evidence that ROS activity actually correlates to degree of DNA damage. In order to define this relationship, we examined our database of patients with male infertility.