Reactive Oxygen Species In Semen Research Articles

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.

O-252 Next generation sequencing analysis of the seminal microbiome in male partners of women with idiopathic recurrent pregnancy loss: results of a prospective cohort study

Abstract Study question Is there an association between the semen microbiome, seminal reactive oxygen species (ROS) and DNA fragmentation in men with recurrent pregnancy loss (RPL)? Summary answer This pilot study outlines the subtle role that microbiota play in influencing ROS and sperm DNA damage for male partners of women with RPL What is known already RPL is defined as the loss of two or more consecutive pregnancies. This devastating condition impacts approximately 1% of couples. Paternal causes are not routinely screened for and an underlying cause is not found in up to 50% of cases. Recent studies have reported an association between elevated seminal ROS and sperm DNA fragmentation in the male partner, and RPL. We hypothesised that seminal microbiota contribute to increased ROS and sperm DNA damage. To test this, we investigated the relationship between seminal bacterial composition and ROS levels in men with proven fertility versus men with a history of RPL. Study design, size, duration We conducted a prospective, case-control study and recruited participants between November 2018 and March 2020 at Imperial College Healthcare NHS Trust. A total of 109 men participated in the study; 46 men with RPL and 63 men with proven fertility and no history of RPL. Each participant attended for a single study visit which consisted of a full medical history, assessment of testicular volume, height, weight, blood samples and production of a semen sample. Participants/materials, setting, methods Routine semen analysis (WHO) and endocrine and lipid profiles were performed for all patients. Semen ROS and DNA fragmentation were performed (luminol and TUNEL methodologies, respectively). ROS were classified as high (>3.77 RLU/s) or low (<3.77 RLU/s). Metataxonomic profiling of samples was performed using Illumina Miseq-based sequencing of the V1-V2 hypervariable regions of bacterial 16S rRNA gene amplicons. Multivariate and univariate modelling was performed to explore associations between metataxonomic profiles, ROS levels and clinical metadata. Main results and the role of chance Men with RPL had higher mean semen volume (p = 0.02) and increased prevalence of high ROS (p = 0.02, Fisher's exact) compared with controls; but other clinical characteristics were similar between groups. A total of 3,700,136 high quality sequence reads were generated for the dataset with an average of 33,946 reads/sample. Hierarchical clustering of bacterial genera relative abundances identified 4 distinct microbial signatures characterised by high relative abundance of 1. Streptococcus, 2. Lactobacillus and Gardnerella, 3. polymicrobial (including Prevotella), and 4. Corynebacterium and Finegoldia. Prevalences of these groups were similar in control and RPL groups (p = 0.11). Additionally, no association between the bacterial genera groups and elevated ROS, DNA fragmentation, or clinical factors such as age, ethnicity, or semen volume were observed (chi-square tests). At species taxonomy level relative abundance of L. crispatus was higher in controls compared to RPL, but did not withstand false discovery rate correction for multiple testing (p = 0.006, q = 0.67). Higher relative abundance of Microbacterium was detected in semen samples with high DNA fragmentation (p = 8.7E-4, q = 0.08). This relationship was even stronger within the RPL cohort (p = 2.8E-5, q = 0.002). No significant enrichment of specific taxa was observed between high or low ROS samples however, low ROS was associated with Corynebacterium relative abundance >20%. Limitations, reasons for caution More patients are required to enhance statistical power. Duplicate sample collection may establish the robustness of seminal compositions observed. Time since last sexual intercourse samples may affect the analysis. Concomitant analysis of the vaginal microbiome of female partners may improve our understanding of how partners may affect each other’s fertility. Wider implications of the findings Our data suggests interactions between microbiota composition, ROS and sperm DNA damage which may be implicated in the pathogenesis of recurrent miscarriage. Further studies are needed to determine if seminal microbiota play causal roles in RPL, and whether interventions modifying the seminal microbiome may modify pregnancy outcomes in affected couples Trial registration number not applicable

Measuring Reactive Oxygen Species in Semen for Male Preconception Care: A Scientist Perspective.

Oxidative stress and elevated levels of seminal and sperm reactive oxygen species (ROS) may contribute to up to 80% of male infertility diagnosis, with sperm ROS concentrations at fertilization important in the development of a healthy fetus and child. The evaluation of ROS in semen seems promising as a potential diagnostic tool for male infertility and male preconception care with a number of clinically available tests on the market (MiOXSYS, luminol chemiluminescence and OxiSperm). While some of these tests show promise for clinical use, discrepancies in documented decision limits and lack of cohort studies/clinical trials assessing their benefits on fertilization rates, embryo development, pregnancy and live birth rates limit their current clinical utility. In this review, we provide an update on the current techniques used for analyzing semen ROS concentrations clinically, the potential to use of ROS research tools for improving clinical ROS detection in sperm and describe why we believe we are likely still a long way away before semen ROS concentrations might become a mainstream preconception diagnostic test in men.

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.

Relevance of Leukocytospermia and Semen Culture and Its True Place in Diagnosing and Treating Male Infertility.

The current WHO 2010 manual for human semen analysis defines leukocytospermia as the presence of peroxidase-positive leukocytes at a concentration >1×106/mL of semen. Granular leukocytes when activated are capable of generating high levels of reactive oxygen species in semen resulting in oxidative stress. Oxidative stress has been correlated with poor sperm quality, increased level of sperm DNA fragmentation and low fertility potential. The presence of leukocytes and pathogens in the semen may be a sign of infection and/or localized inflammatory response in the male genital tract and the accessory glands. Common uro-pathogens including Chlamydia trachomatis, Ureaplasma urealyticum, Neisseria gonorrhoeae, Mycoplasma hominis, and Escherichia coli can cause epididymitis, epididymo-orchitis, or prostatitis. The relationship between leukocytospermia and infection is unclear. Therefore, we describe the pathogens responsible for male genital tract infections and their association with leukocytospermia. The review also examines the diagnostic tests available to identify seminal leukocytes. The role of leukocytospermia in male infertility and its management is also discussed.

Reactive Oxygen Species Secreted by Leukocytes in Semen Induce Self-Expression of Interleukin-6 and Affect Sperm Quality.

Reproductive tract inflammation is considered an important cause of male infertility. Increased leukocytes in semen can produce many reactive oxygen species (ROS), which affect sperm function. The aim of this study is to identify the main source of ROS in seminal plasma and to assess the effect of ROS on leukocytes. Semen samples (n = 20) with leukocyte concentration >1 × 106 were collected from a male infertility clinic. This study mainly compares the sperm function parameters of the normal group and the semen white blood cell group >1 × 106. The results identified that ROS in semen was closely related to sperm function parameters, and CD45+ leucocytes were the main source of ROS. Compared with the control group, the concentration of IL-2, IL-4, IL-6, IFN-γ, and TNF-α was higher in the experimental group. Leukocytes in semen may regulate the secretion of ROS through the mammalian target of rapamycin (mTOR) pathway. A considerable amount of ROS can upregulate the expression of IL-6 in leukocytes via the nuclear factor kappa-B (NF-kB) pathway.

Early and late paternal effects of reactive oxygen species in semen on embryo development after intracytoplasmic sperm injection

ABSTRACTAlthough reactive oxygen species in semen are associated with unfavorable results with respect to assisted reproductive technology, their effects based on the detailed stages of embryo development are unclear. We investigated the relationship between reactive oxygen species in semen and the oocyte fertilization rate, cleavage rate, and blastulation rate of intracytoplasmic sperm injections. This retrospective study enrolled 77 couples who underwent intracytoplasmic sperm injection and analyzed 887 eggs from 141 cycles of intracytoplasmic sperm injection. The reactive oxygen species level in semen was compared between the fertilized and nonfertilized groups, between the good-cleavage-embryo and non-developed-embryo groups, and between the good-quality-blastocyst and poor-quality-blastocyst groups. The cut-off level of reactive oxygen species was calculated to predict good-cleavage-embryo and good-quality-blastocyst development. The fertilization rate was 65.4%, and the mean reactive oxygen species levels were not significantly different between the fertilized and nonfertilized groups. The reactive oxygen species level was significantly higher in the non-developed-embryo group than in the good-cleavage-embryo group (P = 0.0026) and was significantly lower in the good-quality-blastocyst group than in the poor-quality-embryo group (P = 0.015). Cleavage embryos and blastocysts were divided into high- and low-reactive-oxygen-species groups using a cut-off value of 6601 and 4926 relative light units, as calculated from the receiver operating characteristic curve. The rates of good-cleavage embryos and good-quality blastocysts were lower in the high-reactive-oxygen-species group than in the low-reactive-oxygen-species group, which were both statistically significant. To conclude, reactive oxygen species in semen is considered to have an adverse effect on both the early and late stages of embryo development in intracytoplasmic sperm injection.Abbreviations: GnRH, gonadotropin-releasing hormone; ICSI, intracytoplasmic sperm injection; IVF, in vitro fertilization; LPO, lipid peroxidation; NADPH, nicotinamide adenine dinucleotide phosphate; RLU, relative light units; ROC, receiver operating characteristic; ROS, reactive oxygen species

Cancer Chemotherapy and Chemiluminescence Detection of Reactive Oxygen Species in Human Semen.

Advanced treatments have improved the prognosis of cancer survivors. Anticancer drugs generate large amounts of cellular reactive oxygen species (ROS), but their direct effects on sperm ROS production are unclear. We examined 64 semen samples of men who had received cancer chemotherapy, 467 semen samples of men consulting for idiopathic infertility, and 402 semen samples of partners of female patients as a control group. ROS production was calculated as the integrated chemiluminescence between 0 and 200 seconds after the addition of luminol to unwashed semen. We found that their ROS-positive rate of semen samples in the chemotherapy group was significantly higher than that in the control group. We compared the sperm parameters (concentration and motility) and the ROS production levels between chemotherapy subgroups and one of the remaining subgroups with positive ROS, and we found that only sperm motility was significantly lower in the samples in the postchemotherapy subgroup than in the idiopathic infertility subgroup, and that both sperm parameters were significantly lower in those from postchemotherapy subgroup than in the control subgroup. The ROS production level per million spermatozoa in the postchemotherapy subgroup was significantly higher than that in the control subgroup. Additionally, we compared variables, such as age, sperm features, and the duration from the end of the treatment to the first consultation between ROS-positive and ROS-negative subgroups in samples from men in the postchemotherapy group, but we found no significant differences. Of the men in the postchemotherapy group, three underwent a long-term antioxidant therapy, and all of them had low ROS semen levels after that. In conclusion, the production of ROS in semen detected by chemiluminescence from men who undergo cancer chemotherapy is similar to that of men with idiopathic infertility, and long-term oral antioxidant therapy may reduce the amount of ROS in the semen.

The efficacy of oxidation reduction potential(ORP) in male infertility and its relationship with seminal leukocyte concentration

Reactive oxygen species (ROS) in semen has been reported to have negative effect to male fertile capacity, and recent studies reported the efficacy of oxidation-reduction potential (ORP) which reflects the balance of oxidants and antioxidants in semen. The source of ROS in semen is considered as immature spermatozoa and seminal leukocytes, but the detail is still unknown. The aim of this study is to evaluate the relationship between the concentration of seminal leukocytes and oxidative stress level using ROS, ORP. Retrospective study. Between April 2018 and March 2019, 29 infertile males who visited Reproduction Centre of Yokohama City University Medical Center were enrolled. All patients underwent semen analysis and measurement of ROS and ORP levels. The ROS level in semen was measured using Monolight 3010TM Luminometer and the ORP level was measured using MiOXSYS SystemTM. The concentration of peroxidase-positive leukocytes were evaluated using myeloperoxidase staining (Endtz test). The relationship between ROS levels, ORP levels, the concentration of leukocytes and semen parameters were evaluated using correlation analysis. The sperm concentration and motility were 39.6±36.6×106/ml, 30.0±18.9 %, respectively. The total ROS level was 9328.9 (±18851) Relative Light Units, and the ORP level was 46.1(±38.7) mV. The ROS level was significantly correlated with ORP level (r=0.79, p<0.01). The concentration of leukocytes measured by Endtz test was positively correlated with both ORP level (r=0.46, p=0.023) and ROS level (r=0.82, p<0.01). ORP level was negatively correlated with sperm concentration (r= -0.41, p=0.026), while ROS level didn’t show significant correlation with every semen parameters. To our knowledge, this is the first study that showed the significant correlation between ORP levels and seminal leukocytes concentration. Our study suggested that peroxidase-positive leukocyte is one of the main source of ROS in semen. Although ROS was strongly correlated with leukocyte concentration, ORP level is considered to have better potential to reflect the sperm quality.

EFFECT OF NATURAL ORIGIN MEDICINES ON PROOXIDANT-ANTIOXIDANT SYSTEM IN THE PROSTATE RATS AND SPERMATOGENESIS UNDER CONDITIONS OF THE EXPERIMENTAL PROSTATITIS

One of the pathogenic factors in all forms of man's infertility, including the chronic prostatitis as a result, leading to a violation of spermatogenesis, and to the overproduction of reactive oxygen species in semen, which is defined as oxidative stress. The accumulation of reactive oxygen species in sperm leads to damage to sperm membranes, their DNA fragmentation and decrease in their motility. In addition, there is an imbalance of the hormonal profile, which promote the development of man infertility with oxidative stress. The purpose of this work was investigation of prooxidant-antioxidant processes state in rats’ spermogram indicators under conditions of experimental prostatitis and its pharmacological correction by Chondroitin sulfate and its combination with Tribestan and placenta. Non-bacterial prostatitis have been caused by double rectal administration to male rats of 1 ml of the mixture based on a 10 % solution of dimexide in water and turpentine in a 4:1 ratio (model of turpentine prostatitis). Chondroitin sulfate, Chondroitin sulfate + Tribestan, Tribestan, Bioglobin-B, Prostaplant were administered for 14 days. The morphofunctional state of sperm cells and indicators represent of prooxidant-antioxidant processes state in the prostate homogenate have been studied: the content of diene conjugates, lipid hydroperoxides, products which reacts with thiobarbituric acid (TBA products) and reduced glutathione. Turpentine prostatitis is characterized by an intensification of lipid peroxidation processes and decrease of antioxidant protection processes, which negatively affects on the functional state of spermatozoa have been established. All studied medicines have a normalizing effect on the processes of lipid peroxidation and the functional state of spermatozoa. The study of the products of the prooxidant-antioxidant system in homogenates of the prostate gland and rats’ spermogram indicators showed the advantage of using Chondroitin sulfate in compared to other studied medicines.

OR18-5 Elevated Semen Oxidative Stress in Male Partners as Novel Marker of Recurrent Pregnancy Loss

Background: Idiopathic recurrent pregnancy loss (RPL) is the loss of three or more consecutive pregnancies prior to 20 weeks of gestation. Female factors in RPL are well known, but the influence of sperm function on RPL has been poorly understood. Recent studies demonstrate that male partners of women with RPL have higher risk of DNA fragmentation, but the underlying reasons have not been previously studied. Determining the mechanisms of sperm damage has potential to open new therapeutic approaches for couples. Objective: To investigate the role of seminal reactive oxygen species (ROS) as a novel marker of sperm function in male partners of women with RPL. Methods: Serum levels of reproductive hormones and sperm quality of male partners with RPL (N=49) were compared with a group of unaffected controls (n=34). All participants completed a questionnaire for conditions impairing sperm quality such as testicular surgery, chronic use of medication, sexually transmitted infections, smoking and alcohol intake. Semen ROS was measured using a validated in-house chemiluminescent luminol assay. DNA fragmentation was measured using the validated Halosperm G2 kit. Results: Sperm morphology was significantly lower in the RPL group when compared with controls (% sperm with normal morphology: 4.5 ± 0.4, control; 3.4± 0.3, RPL, P<0.001). Mean seminal ROS levels were more than 4-fold higher in the RPL group when compared with controls (mean semen ROS in RLU/sec/106: 2.3 ± 0.9, control; 9.3 ± 4.2, RPL, P<0.05). Mean levels of sperm DNA fragmentation index (DFI) were more than 2-fold higher in the RPL group when compared with controls (mean DFI: 7.4 +1.2, control; 16.3+1.5, RPL, P<0.0001). Levels of serum morning testosterone (T) were non- significantly lower in the RPL group when compared with the control group (mean T in nmol/L: 17.9 ± 1.4, control; 15.8 ± 0.8, RPL, P>0.05). Conclusions: Our data suggest that male partners of women with RPL had elevated levels of semen ROS, elevated sperm DNA fragmentation and lower percentage of normal morphology when compared with controls. Our data have important implications for the management of couples with RPL as male partners of women with RPL may benefit from routine assessment of reproductive endocrine and sperm function.

The association between iron, calcium, and oxidative stress in seminal plasma and sperm quality.

The present study aimed to determine the level of iron and calcium in the seminal plasma of men with different fertility potentials and to examine its relationship with oxidative stress. Seventy-nine sub-fertile patients with asthenoteratozoospermia (AT), n 27; teratoleucozoospermia (TL), n 20; teratozoospermia (Terato), n 32; and 29 healthy donors were included. The ability of spermatozoa to produce reactive oxygen species (ROS) was evaluated by using nitroblue tetrazolium (NBT) staining. The lipid peroxidation end product, malondialdehyde (MDA), and the trace element levels (iron and calcium) were measured spectrophotometrically. Iron and calcium concentrations in seminal plasma of the patient groups were significantly more elevated than the normal group. Nevertheless, both calcium and iron showed strong negative correlations with the total sperm motility and normal sperm morphology, but only iron was positively and significantly associated with multiple anomalies index and seminal leucocyte concentration. On the other hand, the rates of MDA and ROS production in semen were significantly higher in the three abnormal groups than in controls. These two oxidative stress biomarkers were significantly associated with the percentage of atypical forms in semen. However, only semen ROS level was significantly associated with the decreased sperm motility and the sperm leucocytes concentration. Meanwhile, there are positive correlations between seminal iron and calcium content and the studied oxidative stress biomarkers. Oxidative stress and trace element excess are implicated in low sperm quality. Iron and calcium might be the mediators of the effects of oxidative damage and induces lipid peroxidation.

Effects of N-acetyl-cysteine supplementation on sperm quality, chromatin integrity and level of oxidative stress in infertile men

BackgroundInfertile men have higher levels of semen reactive oxygen species (ROS) than fertile men. High levels of semen ROS can cause sperm dysfunction, sperm DNA damage and reduced male reproductive potential. This study investigated the effects of supplementation with N-acetyl-cysteine (NAC) on the sperm quality, chromatin integrity and levels of oxidative stress in infertile men.MethodsThe study was carried out in the unit of ACECR Infertility Research Center, Qom, Iran. The patients consisted of 50 infertile men with asthenoteratozoospermia who received NAC (600 mg/d) orally for 3 months, after which they were compared with pre-treatment status. Semen was analyzed according to WHO (2010), followed by the assessment of protamine content [chromomycin A3 (CMA3)] and DNA integrity [terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling (TUNEL)]. Oxidative stress markers, i.e. total antioxidant capacity (TAC) and malondialdehyde (MDA), as well as hormonal profile (LH, FSH, Testosterone and Prolactin) were determined by ELISA kit.ResultsAfter NAC treatment, patients’ sperm count and motility increased significantly whereas abnormal morphology, DNA fragmentation and protamine deficiency showed significant decreases compared to pre-treatment levels (P < 0.05). Hormonal profile improvement was associated with lowered FSH and LH levels and increased amount of testosterone (P < 0.05). TAC significantly increased and MDA decreased with an inverse significant correlation between TAC and MDA (P < 0.05).ConclusionNAC oral supplementation may improve sperm parameters and oxidative/antioxidant status in infertile males.

Reduced Testicular Steroidogenesis and Increased Semen Oxidative Stress in Male Partners as Novel Markers of Recurrent Miscarriage.

Recurrent pregnancy loss, (RPL) affecting 1%-2% of couples, is defined as ≥3 consecutive pregnancy losses before 20-week' gestation. Women with RPL are routinely screened for etiological factors, but routine screening of male partners is not currently recommended. Recently it has been suggested that sperm quality is reduced in male partners of women with RPL, but the reasons underlying this lower quality are unclear. We hypothesized that these men may have underlying impairments of reproductive endocrine and metabolic function that cause reductions in sperm quality. After ethical approval, reproductive parameters were compared between healthy controls and male partners of women with RPL. Semen reactive oxygen species (ROS) were measured with a validated inhouse chemiluminescent assay. DNA fragmentation was measured with the validated Halosperm method. Total sperm motility, progressive sperm motility, and normal morphology were all reduced in the RPL group vs controls. Mean ±SE morning serum testosterone (nmol/L) was 15% lower in RPL than in controls (controls, 19.0 ± 1.0; RPL, 16.0 ± 0.8; P < 0.05). Mean ±SE serum estradiol (pmol/L) was 16% lower in RPL than in controls (controls, 103.1 ± 5.7; RPL, 86.5 ± 3.4; P < 0.01). Serum luteinizing hormone and follicle-stimulating hormone were similar between groups. Mean ±SE ROS (RLU/sec/106 sperm) were 4-fold higher in RPL than in controls (controls, 2.0 ± 0.6; RPL, 9.1 ± 4.1; P < 0.01). Mean ±SE sperm DNA fragmentation (%) was 2-fold higher in RPL than in controls (controls, 7.3 ± 1.0; RPL, 16.4 ± 1.5; P < 0.0001). Our data suggest that male partners of women with RPL have impaired reproductive endocrine function, increased levels of semen ROS, and sperm DNA fragmentation. Routine reproductive assessment of the male partners may be beneficial in RPL.

Seminal plasma cell-free mitochondrial DNA copy number is associated with human semen quality

ObjectivePrevious studies have shown that mitochondrial DNA (mtDNA) copy number in human sperm is associated with semen quality. This study examines whether there is association between seminal cell-free mtDNA copy number and human semen parameters. Study designSeminal cell-free mtDNA copy number was measured in 205 men. Semen reactive oxygen species (ROS) level was also measured. Sperm quality was assessed using World Health Organization criteria, while sperm motility parameters were determined by computer-aided sperm analysis. ResultsComparing with normozoospermia, patients with asthenozoospermia and oligoasthenozoospermia had a significant decrease in cell-free mtDNA copy number and an increase in ROS level in seminal plasma (P < 0.05 for all). Seminal cell-free mtDNA copy number positively correlated with sperm concentration, motility, morphology and motion characteristics (r for 0.247 to 0.673, P < 0.05 for all). Semen ROS level negatively correlated with sperm concentration, motility, and motion characteristics (r for −0.261 to −0.676, P < 0.05 for all). There is a negative relationship between seminal cell-free mtDNA copy number and ROS level (r= -0.573, P < 0.05). ConclusionThe results suggested that seminal cell-free mtDNA copy number is associated with semen parameters, which may serve as a novel diagnostic marker of semen quality.

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 ...

Negative correlation between presence of reactive oxygen species and Sperm Motility Index in whole semen samples of infertile males

ObjectiveTo investigate the relationship between the Sperm Motility Index and the presence of reactive oxygen species in semen. MethodsWe retrospectively analyzed Sperm Motility Index measured by the Sperm Quality Analyzer and reactive oxygen species levels in 92 semen samples of infertile male patients who visited the Ebina Ladies’ Clinic between September 2011 and June 2012. Using the same semen samples, we also analyzed 8 sperm parameters measured by computer-assisted semen analysis and validated the relationship with the Sperm Motility Index. ResultsThe presence of reactive oxygen species in semen was positive in 19 samples and negative in 73 samples. In the reactive oxygen species-positive group, there was a significant negative correlation between the logarithm of reactive oxygen species level and Sperm Motility Index (p = 0.039). ConclusionsThis is the first study to find a significant negative correlation between Sperm Motility Index and reactive oxygen species level. This result indicates that the presence of reactive oxygen species in semen may inhibit the fertilization ability of spermatazoa as measured by the Sperm Motility Index.

Inflammatory and anti-inflammatory cytokines in the seminal plasma of infertile men suffering from varicocele.

Varicocele is one of causes of the declined sperm quality and low sperm production, which can lead to infertility in males. There are several experimental and epidemiological findings which support the idea that inflammatory mechanisms play an essential role in varicocele pathogenesis. Besides, in this pathological state, interleukin-37 (IL-37) as an anti-inflammatory cytokine is able to bind interleukin-18-binding protein (IL-18BP), and subsequently binds IL-18 receptor β, inhibiting the pro-inflammatory activity of IL-18. To explore the interaction between IL-37 and IL-18 in infertility, we measured the amount of these cytokines in the seminal fluid of infertile men affected by varicocele. Theseminal plasma levels of IL-37 and IL-18 were measured in 75 infertile men with varicocele and 75 healthy fertile controls (age range, 30-48years) using enzyme-linked immunosorbent assay. The seminal levels of IL-37 and IL-18 were significantly increased in infertile men with varicocele when compared to fertile controls (p<.0001). Because of the essential role(s) of cytokines in inflammatory response of cell systems, it could be possible that sperm motility is reduced following increased IL-18, activated neutrophils and reactive oxygen species in semen of infertile patients with varicocele. Moreover, the results of this study indicated that interaction between IL-37 and IL-18Rβ can lead to reduced inflammatory responses. It seems that IL-37 might be a potential biomarker and therapeutic target for male infertility.