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Micro-organisms are tightly integrated into host-microbiota ecosystem. Microbiota of human semen has been studied so far mostly in case of infertility or prostatitis. We aimed to reveal possible impact of sexual debut on seminal microbiota in healthy young men. The study group included 68 young healthy men, of them 12 men without sexual experience, 11 men with single lifetime sexual partner and 45 men with multiple lifetime sexual partners. Basic semen parameters were similar for all subgroups, and no correlation between sexual experience and WBC counts in semen was found. A man could harbour one to nine different bacteria in his semen; the total concentration of bacteria ranged from 2.3 to 7.3 log10 CFU/mL of semen. Lower total bacterial concentration and lower bacterial diversity was observed in men without sexual experience than in sexually experienced men (p < 0.05), with significant positive correlation between these two parameters (r = 0.54; p < 0.0001). In conclusion, the sexual debut is associated with the enrichment of seminal microbiota but not with the influx of WBC or changes in basic seminal parameters.

To compare markers of semen quality and related reproductive indicators in middle-aged males in relation to serum prostate-specific antigen (PSA) levels.A total of 384 voluntary middle-aged men who underwent screening for prostate health were recruited. Reproductive function, semen quality, hormonal parameters, prostate-related pathologies, and PSA levels were measured.Total sperm count and sperm concentration were decreased in men with elevated serum PSA compared with age-matched controls. Serum FSH levels differed significantly in subjects with non-malignant, pre-malignant, and malignant prostate conditions. PSA levels in men with three normal semen variables (semen volume ≥ 1.5 mL, sperm density ≥ 15 × 10(6)/mL, and A + B motility ≥ 40%) were significantly lower compared with subjects with altered parameters (1.51 ng/ml vs. 2.47 ng/ml, respectively, p = 0.002). PSA showed a negative correlation with semen volume and total sperm count, and a positive correlation with semen interleukin-6.Our data demonstrate that serum PSA levels are related to impaired reproductive parameters in middle-aged subjects. While there is no consensus about the potential link between male factor infertility, impaired reproductive indicators, and prostate pathologies, this topic needs additional research focusing on the interactions between male aging, reproductive function, and the prostate.

We investigated the prevalence of oxidative stress in male partners of infertile couples displaying different inflammation patterns in their genital tract and/or oligospermia. The levels of acknowledged oxidative stress markers (8-isoprostanes [8-EPI], diene conjugates, reactive oxygen species-total antioxidant capacity [ROS-TAC] score) were elevated in both leukocytospermic men and subjects whose inflammation was limited only to expressed prostatic secretion and/or post-massage urine. Oligospermic men with severe inflammation were different from oligospermic men who had no inflammation at all - the former had elevated 8-EPI, diene conjugates and ROS-TAC score when compared to the latter indicating that inflammation has substantially more significant impact on oxidative stress markers than oligospermia status. At the same time nitric oxide (NO) and hydrogen peroxide (H2 O2 ) levels were significantly increased not only in the men with severe inflammation but also in men with borderline inflammation in their genital tact and in men having non-inflammatory oligospermia. NO, H2 O2 and 8-EPI were negatively correlated with per cent of normal sperms, and NO and H2 O2 showed negative correlation also with sperm count. We can conclude that in men presenting with couple infertility the acknowledged oxidative stress markers are substantially associated with markers of inflammation in genital tract while NO and H2 O2 display high levels also in patients with mild inflammation and non-inflammatory oligospermia.

Purpose We aimed to compare the associations between semen quality, associated reproductive indicators and the main prostate-related parameters in middle-aged men. Materials and Methods: This is a prospective study on 422 middle-aged men who underwent the screening for prostate health. Their reproductive function, semen quality and prostate-related pathologies were investigated. Results Significant associations between semen quality and prostate-related parameters could be seen. Total sperm count and sperm density decreased along with the increase of the I-PSS score and total prostate volume. Also, the related lower urinary tract characteristics showed a negative correlation with main semen parameters for all investigated subjects. No significant differences in age, testicular size, and hormonal parameters were found between the subjects with or without lower urinary tract symptoms and prostate enlargement. Conclusions Our study suggests that altered seminal parameters in middle-aged men are associated with LUTS, prostate enlargement and/or bladder outlet obstruction. Although the assessments of prostate and lower urinary tract symptoms may not replace the semen parameters evaluating the male reproductive status, there is a need for further and more detailed investigations about the pathways behind these associations as well as possible related conditions.

Objectives To profile the seminal microbiome applying next generation sequencing. Methods Semen samples of 67 men were involved in the study (21 men with and 46 men without prostatitis). Seminal microbiomes were profiled applying the method that uses combinatorial sequence tags attached to polymerase chain reaction primers that amplify the ribosomal ribonucleic acid V6 region. Amplified polymerase chain reaction products were sequenced using an Illumina paired-end protocol on HiSeq2000 platform. Results The most abundant phylum in semen was Firmicutes, comprising nearly half of the sequences found (median 41.7%, quartiles 28.5–47.2%) followed by Bacteroidetes, Proteobacteria and Actinobacteria. The counts of lactobacilli were higher in healthy men than prostatitis patients (27% [20.2–34.6%] vs 20.2% [4.9–25.0%]; P = 0.05), especially for Lactobacillus iners. Proteobacteria comprised higher proportions in prostatitis patients than healthy men. The species richness was higher in prostatitis patients than healthy men (inverted Simpson index 13.5 ± 5.8 vs 10.3 ± 4.0). Conclusions The semen of chronic prostatitis patients contains fewer health-supporting lactobacilli, and has higher species diversity than that of healthy men. Firmicutes (especially lactobacilli), Bacteroidetes, Proteobacteria and Actinobacteria comprise the highest proportion of seminal microbiome.

Context: The detailed role of FSH in contributing to male testicular function and fertility has been debated. We have previously identified the association between the T-allele of the FSHB promoter polymorphism (rs10835638; G/T, −211 bp from the mRNA start) and significantly reduced male serum FSH. Objective: In the current study, the T-allele carriers of the FSHB −211 G/T single nucleotide polymorphism represented a natural model for documenting downstream phenotypic consequences of insufficient FSH action. Design and Subjects: We genotyped rs10835638 in the population-based Baltic cohort of young men (n = 1054; GG carriers, n = 796; GT carriers, n = 244; TT carriers, n = 14) recruited by Andrology Centres in Tartu, Estonia; Riga, Latvia; and Kaunas, Lithuania. Marker-trait association testing was performed using linear regression (additive, recessive models) adjusted by age, body mass index, smoking, and recruitment center. Results: Serum hormones directly correlated with the T-allele dosage of rs10835638 included FSH (additive model, P = 1.11 × 10−6; T-allele effect, −0.41 IU/liter), inhibin-B (P = 2.16 × 10−3; T-allele effect, −14.67 pg/ml), and total testosterone (P = 9.30 × 10−3; T-allele effect, −1.46 nmol/liter). Parameters altered only among TT homozygotes were reduced testicular volume (recessive model, P = 1.19 × 10−4; TT genotype effect, −9.47 ml) and increased serum LH (P = 2.25 × 10−2; TT genotype effect, 1.07 IU/liter). The carrier status of rs10835638 alternative genotypes did not affect sperm motility and morphology, calculated free testosterone, serum SHBG, and estradiol concentrations. Conclusion: We showed for the first time that genetically determined low FSH may have wider downstream effects on the male reproductive system, including impaired testes development, altered testicular hormone levels (inhibin-B, total testosterone, LH), and affected male reproductive potential.

The purpose of the current study was to evaluate the importance of androgen receptor (AR) gene haplotypes and polymorphic CAG/GGN microsatellites in the aetiology of male infertility. We genotyped six haplotype-tagging single nucleotide polymorphisms and CAG/GGN microsatellites of the AR gene in 112 infertile and 212 control Estonian men. A total of 13 AR haplotypes (HAP1-13) were identified, among which HAP4 was found to confer increased risk for male infertility (OR = 5.15, 95% CI = 1.75-15.15, p = 0.003). However, infertile patients and controls had similar lengths and distributions of both AR CAG (mean +/- SD number of repeats 21.1 +/- 2.5 vs. 21.2 +/- 2.3, respectively) and GGN (mean +/- SD number of repeats 22.5 +/- 1.5 vs. 22.4 +/- 1.9, respectively) repeats. In addition, HAP2 was associated with more CAG repeats (r = 1.17, p = 0.033) and HAP3 with fewer CAG repeats (r = -2.93, p < 0.001) than the major haplotype HAP1. HAP3 and HAP4 were associated with more GGN repeats (r = 1.35, p = 0.001 and r = 1.36, p = 0.002, respectively) than HAP1. In conclusion, our results implicated the AR-HAP4 gene haplotype in increased risk for male infertility, while no association was found between AR CAG/GGN microsatellites and impaired spermatogenesis.

BACKGROUND No polymorphisms affecting serum FSH levels have been described in the human FSHB gene. We have identified a potential regulatory single nucleotide polymorphism (SNP, rs10835638; G/T) 211 bp upstream from the FSHB mRNA transcription start-site, located within a highly conserved region among placental mammals. We aimed to determine the correlation of carrier status of rs10835638 alternative alleles with serum FSH level in men, and testicular and hormonal parameters. METHODS A quantitative genetic association study using a cohort of healthy men (n = 554; age 19.2 ± 1.7 years) visiting the Centre of Andrology, Tartu University Hospital, Estonia. RESULTS Rs10835638 (allele frequencies: G 87.6%, T 12.4%) was significantly associated with serum FSH level (analysis of variance: F = 13.0, P = 0.0016, df = 1; regression testing for a linear trend: P = 0.0003). Subjects with the GG genotype exhibited higher FSH levels (3.37 ± 1.79 IU/l, n = 423) compared with heterozygotes (2.84 ± 1.54 IU/l, n = 125) (P = 0.0005), the group of T-allele carriers (GT+TT, 2.78 ± 1.51 IU/l, n = 131) (P = 0.0005) and TT-homozygotes (2.02 ± 0.81 IU/L, n = 6) (P = 0.031). Rs10835638 was also associated with significant (P < 0.05) reduction in free testosterone index and testes volume, but increased semen volume, sex hormone-binding globulin, serum testosterone and estradiol. LH and inhibin-B levels did not differ significantly between groups. CONCLUSIONS The identification of a regulatory SNP in FSHB promoter paves the way to study the effect of constitutively low FSH on male health and fertility. As FSH contributes to follicular development and sex steroid production in women, the role of this FSHB variant in female reproductive success is still to be addressed.

Purpose The purpose of this research was to evaluate the effect of a specific fermented whey product on lower urinary tract symptoms, main prostate related indices and oxidative stress/inflammatory markers in urine and seminal plasma in men with moderate dysuric symptoms. An additional purpose was to clarify associations between different parameters with special emphasis on pain. Methods This was a prospective randomized double-blind 4-weeks study on men with moderate lower urinary tract symptoms who underwent the evaluation for quality of life at the baseline and at the end of the study. The symptoms were characterized by International Prostate Symptom Score (I-PSS) and National Institutes of Health Chronic Prostatitis Symptom Index (NIH-PSI), the maximum urinary flow and the main prostate-related indices. In order to obtain more comprehensive information about the effects of fermented whey product on systemic oxidative stress marker 8-EPI and seminal plasma inflammatory markers (interleukin-6 and interleukin-8) were also measured. Results After 4 weeks consumption of fermented whey product there was a statistically significant decrease of prostate-specific antigen level in serum and systemic stress marker 8-EPI in urine compared to control group. Maximum urinary flow and NIH-PSI all studied scores and sub-scores had also significant improvement. In addition, seminal plasma interleukin-8 level substantially decreased. Conclusions The consumption of special fermented whey product improved urinary function, reduced lower urinary tract symptoms, systemic oxidative stress marker and seminal plasma inflammatory status. Thus it contributed to an improvement of the quality of life in men with moderate lower urinary tract symptoms.

To compare the associations between non-malignant prostate conditions, genital tract inflammation, and reproductive function in middle-aged men.Three-hundred and eighty-two voluntary male subjects who underwent the screening for prostate health were recruited for the study. Semen quality and associated reproductive indicators, seminal inflammation, and prostate-related pathologies were evaluated.Sperm motility and prostate-related parameters were significantly impaired in patients with chronic prostatitis syndromes and lower urinary tract symptoms in comparison with controls. Elevated seminal markers of inflammation were in positive association with body mass index, prostate-specific antigen, and estradiol level in serum while in negative association with semen volume, total sperm count, and sperm motility. According to WHO reference limits, speculative cutoff values for WBC and IL-6 in semen to detect reduced sperm parameters were 0.342 M/mL and 56.8 ng/L, respectively.According to our data, one of the possible pathways for impaired reproductive quality in male subjects45 years could be related to infection and inflammation in the genital tract with subsequent (partial) obstruction and damage of prostate and other male accessory glands.

The aim of this study was to compare the reproductive parameters and the health-related, lifestyle and educational factors in middle-aged healthy men and male partners of infertile couples. Our patient group included 164 male partners of infertile couples with a preceding period of infertility of at least 12 months and 61 men attending a prostate health screening and considering themselves healthy. Significant differences between the groups were found in testicular volume, total sperm output, density and morphology as well as oestradiol levels in serum. The analysis showed significant positive correlations between testicular volume and semen quality, while negative correlations were observed between gonadotrophin levels and sperm parameters in both groups. Physical activity and sexual capability were higher in healthy men, while coital frequency and a history of sexually transmitted diseases were higher in male partners of infertile couples. The impact of physical activity and sexual capability on semen quality for all subjects was revealed. We can conclude that impaired reproductive function, that is, semen quality, in men >45 years is related not only with general male ageing but obvious differences between subjects of infertile couples and healthy middle-aged men can be seen. Their relations with lifestyle, environmental or physiological factors need further study.

Klinefelter’s syndrome (KS) is the most common human sex chromosome disorder, with a prevalence of 1 in 660 men. The phenotype is variable, but the most constant findings are small and firm testes, decreased testosterone level, infertility, eunuchoid body proportion, increased height, and learning disabilities, due to the presence of extra X chromosome(s) (Smyth and Bremner 1998). KS patients have frequently other diseases of lung, skin, liver and kidney (Morales et al. 1992; Swerdlow et al. 2005). Restrictive lung defects have been attributed to chest wall abnormalities, decreased respiratory muscle strength and increased chest wall compliance (Huseby and Petersen 1981). The likely cause is, decrease of lung compliance due to diminished elasticity of the lung matrix, but its biochemical cause is unknown (Morales et al. 1992). One of the causes could be the mutations in the alpha-1 antitrypsin (AAT) gene (SERPINE1), which is highly polymorphic, with more than 100 alleles identified so far. The alleles are categorized into normal, deficient and null variants on the basis of the plasma level and function of AAT. The protein phenotype is classified according to the ‘Pi’ (protease inhibitor) system, as defined by plasma isoelectric focusing. Normal Pi M types account for 95% of alleles in Caucasian individuals and are characterized by normal plasma levels. Pi X is a rare normal allele variant. Pi Z, S and null types are the most frequent AAT deficiency variants described in lung pathology, but Pi ZZ is associated with both pulmonary and liver diseases (Kaczor et al. 2007; Greene et al. 2008). AAT gene is located on chromosome 14q32.1 (Schroeder et al. 1985).

Several studies have suggested the association of disturbed genital tract microbiota with infertility. Our aim was to clarify the influence of sexual intercourse on partner’s genital tract microbiota in infertile couples. Seventeen couples were studied, and in 5 men inflammatory prostatitis (IP) was diagnosed. Semen samples were collected during menstruation of the female counterpart, two self-collected vaginal samples were taken 3–5 days later – before intercourse and 8–12 h after intercourse. Ureaplasma parvum was found in 59% of women, its prevalence was higher in women whose partner had IP, as well as in half of their male partners. Sexual intercourse caused significant shifts in vaginal microbiota – increase of Nugent score and shifts in cultured microbiota (emergence and disappearance of several species). These changes were less expressed in the presence of normal vaginal microbiota but more prominent in the partners of IP men. These changes may interfere with fertilization.

Objectives To determine the possible predictive values of seminal interleukin-6 (IL-6) and serum prostate-specific antigen (PSA), as well as their combined values, in differentiating between subjects with or without asymptomatic inflammatory prostatitis. Methods The study group consisted of 490 men (mean age 18.9 ± 1.8 years, range 16-25). Cytologic examination of all ejaculates (using Bryan-Leishman-stained slides) and clinical examination for possible pathologic findings in the genital region were performed. The subjects with any clinical symptoms of inflammation were excluded. The levels of PSA in the blood serum and IL-6 in the seminal plasma were also determined. The IL-6 and PSA levels for different leukocytospermia status were statistically compared, and receiver operating characteristic curves were designed to determine the sensitivity versus specificity and the positive and negative predictive values of IL-6 and PSA levels against different thresholds of leukocytospermia (0.2, 0.5, and >1.0 × 10 6 leukocytes/mL). Results The levels of both IL-6 in the seminal plasma and PSA in the blood serum were significantly greater in National Institutes of Health prostatitis IV than in the controls. The receiver operating characteristic curves for seminal IL-6 and serum PSA showed high negative prognostic values for all 3 leukocytospermic subgroups, and positive prognostic values were seen only with IL-6 in the lower leukocytospermic range. Conclusions Both seminal IL-6 and serum PSA are excellent negative predictive markers for asymptomatic inflammatory prostatitis in young men, although positive predictive values of these biomarkers remain less indicative in this age group.

Context: The human FSHB promoter polymorphism (rs10835638; −211 G/T) has been associated with serum FSH in a cohort of young Estonian men. The minor allele carriers had reduced serum FSH (15.7% in GT heterozygotes; 40% in TT homozygotes) compared with GG homozygotes. Objective: Because FSH is essential for normal spermatogenesis and fertility, we speculated that abnormalities in FSH action could contribute to male infertility. We sought to study whether genetically inherited constitutively reduced FSH levels may affect male reproduction and replicate the association between rs10835638 and serum FSH among infertile male patients. Design: Genotyping of rs10835638 in a cohort of infertile men (n = 1029; Andrology Center of the Tartu University Clinics, Estonia), including idiopathic infertility cases (IIFC; n = 750). Patients: Patients included male partners (sperm concentration

Objectives To determine the prevalence of asymptomatic inflammatory (National Institutes of Health category IV) prostatitis in a cross-sectional community sample of young men. Methods The study group consisted of 565 men aged 18.9 ± 1.8 years (mean ± standard deviation). Cytologic examination of all ejaculates (using Bryan-Leishman stained slides), as well as analysis for basic semen parameters (volume, concentration, and motility) and clinical examination for possible pathologies in the genital region, were performed. Subjects with any clinical symptoms of inflammation were excluded. Levels of prostate-specific antigen in blood serum and interleukin-6 in seminal plasma were determined as well. Results The prevalence of asymptomatic inflammatory prostatitis (greater than 1 × 10 6 white blood cells [WBC]/mL in semen, according to World Health Organization guidelines) was 6.0%, but when we used a lower threshold suggested by our previous studies (greater than 0.2 × 10 6 WBC/mL), the prevalence was 19.0%. In this study the analysis did not show any significant effect of leukocytospermia on sperm quality, except higher sperm concentration in subjects with moderate leukocytospermia (0.2–1 × 10 6 WBC/mL). We did not detect any seasonal differences in the prevalence of asymptomatic inflammatory prostatitis. Interleukin-6 and prostate-specific antigen levels were significantly higher in leukocytospermic subjects than in those without leukocytospermia. Conclusions Asymptomatic inflammatory prostatitis has a notable prevalence among healthy young men, suggesting the need for further studies to investigate pathogenetic mechanisms of the disease. Lack of negative effect of leukocytospermia on basic semen parameters may indicate higher functional reserve of accessory sex glands in this age group.