Subfertile Men Research Articles

High prevalence of hypogonadism and associated impaired metabolic and bone mineral status in subfertile men.

It is yet unknown to which degree young subfertile men present with signs of hypogonadism and whether low testosterone concentration, like in older men, is associated with risk of osteoporosis and metabolic derangements in those subjects. The objective was therefore to investigate the prevalence of hypogonadism and its association with metabolic and bone parameters in young subfertile men. Cross-sectional case-control study. Men from infertile couples (n = 192); 18-50 years; sperm concentration <20 × 10(6) /ml and population-based age-matched controls (n = 199). Blood sampling, anthropometrics, blood pressure, ankle-brachial index and assessment by dual-energy X-ray absorptiometry were undertaken. Odds ratios of biochemical hypogonadism (total testosterone <8·0 nmol/l and/or luteinizing hormone ≥8·6 IU/l and/or ongoing androgen replacement therapy) were calculated. Serum concentrations of sex hormones, lipids, glucose, insulin and HbA1c were assessed and bone mineral density (BMD) evaluated. Compared to controls, the risk of hypogonadism was increased among subfertile men (OR 10; 95% CI, 5·1, 22), being highest in those with nonobstructive azoospermia. Hypogonadal men had higher HbA1c concentration (mean diff. 2·8 mmol/mol; 95% CI, 0·64, 4·9; P = 0·011) and lower lumbar spine BMD (mean diff. 0·05 g/cm(2) ; 95% CI, 0·01, 0·10; P = 0·032) compared to eugonadal subfertile men, even more pronounced in subfertile men with subnormal testosterone levels. Young subfertile men have 10 times increased OR of hypogonadism, which is linked to increased HbA1c and decreased bone mineralization. Endocrine assessment and, if needed, measures to prevent metabolic sequelae and osteoporosis should be included in the routine management of men from infertile couples.

The Sperm Chromatin Structure Assay (SCSA®) and other sperm DNA fragmentation tests for evaluation of sperm nuclear DNA integrity as related to fertility

Thirty-five years ago the pioneering paper in Science (240:1131) on the relationship between sperm DNA integrity and pregnancy outcome was featured as the cover issue showing a fluorescence photomicrograph of red and green stained sperm. The flow cytometry data showed a very significant difference in sperm DNA integrity between fertile and subfertile bulls and men. This study utilized heat (100°C, 5min) to denature DNA at sites of DNA strand breaks followed by staining with acridine orange (AO) and measurements of 5000 individual sperm of green double strand (ds) DNA and red single strand (ss) DNA fluorescence. Later, the heat protocol was changed to a low pH protocol to denature the DNA at sites of strand breaks; the heat and acid procedures produced the same results.SCSA data are very advantageously dual parameter with 1024 channels (degrees) of both red and green fluorescence. Hundreds of publications on the use of the SCSA test in animals and humans have validated the SCSA as a highly useful test for determining male breeding soundness. The SCSA test is a rapid, non-biased flow cytometer machine measurement providing robust statistical data with exceptional precision and repeatability. Many genotoxic experiments showed excellent dose response data with very low coefficient of variation that further validated the SCSA as being a highly powerful assay for sperm DNA integrity.Twelve years following the introduction of the SCSA test, the terminal deoxynucleotidyl transferase-mediated fluorescein-dUTP nick end labelling (TUNEL) test (1993) for sperm was introduced as the only other flow cytometric assay for sperm DNA fragmentation. However, the TUNEL test can also be done by light microscopy with much less statistical robustness.The COMET (1998) and Sperm Chromatin Dispersion (SCD; HALO) (2003) tests were introduced as light microscope tests that don’t require a flow cytometer. Since these tests measure only 50–200 sperm per sample, they suffer from the lack of the statistical robustness of flow cytometric measurements. Only the SCSA test has an exact standardization of a fixed protocol. The many variations of the other tests make it very difficult to compare data and thresholds for risk of male factor infertility. Data from these four sperm DNA fragmentation tests plus the light microscope acridine orange test (AOT) are correlated to various degrees.

Relationship between Lipids Levels of Serum and Seminal Plasma and Semen Parameters in 631 Chinese Subfertile Men.

ObjectiveThis prospective study was designed to investigate the relationship between lipids levels in both serum and seminal plasma and semen parameters.Methods631 subfertile men were enrolled. Their obesity-associated markers were measured, and semen parameters were analyzed. Also, seminal plasma and serum TC, TG, HDL and LDL and serum FFA, FSH, LH, total testosterone (TT), estradiol (E2) and SHBG levels were detected.ResultsSeminal plasma and serum TG, TC and LDL levels were positively related to age. Serum TC, TG and LDL were positively related to obesity-associated markers (P < 0.001), while only seminal plasma TG was positively related to them (P < 0.05). For lipids levels in serum and seminal plasma, only TG level had slightly positive correlation between them (r = 0.081, P = 0.042). There was no significant correlation between serum lipids levels and semen parameters. However, seminal plasma TG, TC, LDL and HDL levels were negatively related to one or several semen parameters, including semen volume (SV), sperm concentration (SC), total sperm count (TSC), sperm motility, progressive motility (PR) and total normal-progressively motile sperm counts (TNPMS). Moreover, seminal plasma TG, TC, LDL and HDL levels in patients with oligospermatism, asthenospermia and teratozoospermia were higher than those with normal sperm concentration, motility or morphology. After adjusting age and serum LH, FSH, TT, E2 and SHBG levels, linear regression analysis showed that SV was still significantly correlated with seminal plasma LDL (P = 0.012), both of SC and TSC with seminal plasma HDL (P = 0.028 and 0.002), and both of PR and sperm motility with seminal plasma TC (P = 0.012 and 0.051).ConclusionThe abnormal metabolism of lipids in male reproductive system may contribute to male factor infertility.

Summary evidence on the effects of varicocele treatment to improve natural fertility in subfertile men.

The objective of this review was to summarize the evidence concerning the benefit of varicocele treatment to improve natural fertility in subfertile males. We also analyzed the effect of varicocele treatment on conventional semen parameters and sperm functional tests. An electronic search to collect the data was performed using the PubMed/MEDLINE databases until July 2015. Data pooled from a variety of study designs indicate that varicocelectomy improves semen parameters in the majority of the treated men with clinical varicocele and abnormal semen parameters regardless of the chosen surgical method. Surgical varicocele repair was beneficial not only for alleviating oxidative stress-associated infertility but also to improve sperm nuclear DNA integrity. However, given the low magnitude of the effect size in sperm DNA integrity, further research is needed to elucidate its clinical significance. Conflicting results on the effect of varicocele treatment on natural fertility seem to be due to heterogeneous study designs and, more importantly, patient selection criteria. When these issues are controlled, current evidence indicates that treatment of subclinical varicocele is not warranted, as it does not seem to improve fertility. On the contrary, fair evidence indicates that varicocele treatment should be offered to infertile patients with palpable varicocele and abnormal semen parameters. This evidence supports the current guidelines issued by the American Urological Association and European Association of Urology, which state that varicocele treatment should be offered to male partners of infertile couples presenting for evaluation with clinical varicocele and semen parameters alterations.

Pattern of Changes in Semen Characteristics in Subfertile Males of South Asian Subcontinent: Analysis of 1,000 Semen Samples

ABSTRACT Aim To investigate and compare whether semen quality has changed in South Asian subfertile males attending three different infertility units. Objectives To characterize the semen parameters of subfertile men in South Asian region and to study the prevalence of abnormal semen parameters of subfertile males of South Asian subcontinent. Materials and methods Study period — January 2014 to December 2014. Setting: Centre for Assisted Reproduction, BIRDEM, Dhaka, Bangladesh Malhotra Nursing Home, Agra, Uttar Pradesh, India Lahore Institute of Fertility and Endocrinology, Lahore, Punjab, Pakistan The possible decline in semen parameters related to male fertility was evaluated by determining the volume, sperm count, motility, viability, and morphology in individuals attending infertility clinics. WHO 2004 initially and then WHO 2010 criteria were used to analyze the semen sample. Results There was no significant age difference among the participants from three different countries. There was significant increase rate of azoopsermia in all the population groups especially in Bangladeshi population. Necrozospermia (100%) immotile sperm is higher in Pakistani populations compare to Bangladeshis and Indians. Normal sperm parameters (Normozoospermia) rate is compatible in Pakistani and Indian male but lower rate is noted in Bangladeshi population. No significant changes were noted in oligozospermia group but significant changes in asthenozospermia noted in Bangladeshi population compare two other two countries Statistical analysis Statistical analysis were performed by using Statistical Software Package (SAS) version 9.1.3 (SAS Institute Inc., Cary, NC, USA). Conclusion Our finding suggested that the values of sperm parameters were in agreement with WHO criteria, significantly different in subfertile men of this region. A further study with good quality collaborative long term research should be carried out, including aspects such as semen quality, reproductive hormones and xenobiotics as well as a strict definition of fecundity in this region is needed. How to cite this article Mahmud N, Malhotra N, Malhotra J, Khan HL. Pattern of Changes in Semen Characteristics in Subfertile Males of South Asian Subcontinent: Analysis of 1,000 Semen Samples. J South Asian Feder Obst Gynae 2016;8(2):145-148.

Semen characteristics in consecutive ejaculates with short abstinence in subfertile males

This study reports the favourable semen characteristics of 73 subfertile oligozoospermic men with short abstinence periods up to 40 min. Semen characteristics were compared between initial and consecutive ejaculate showing improved semen parameters: progressive grade A spermatozoa, morphology and sperm concentration. Median concentrations in initial and consecutive ejaculates were 10 million/ml and 17 million/ml, respectively. The second sample had a higher median normal morphology (7% versus 6%, P < 0.001). The median of non-progressive spermatozoa (Grade C) was significantly lower in the consecutive sample than the initial sample (0% versus 5%, P < 0.01). Medians for slow progression spermatozoa (B grade) and immotile spermatozoa (D grade) were lower in the consecutive samples (20% versus 13%, P < 0.01 and 60% versus 50%, P < 0.001, respectively). The median for rapid motility (Grade A) was significantly higher in the consecutive sample than the first (30% versus 5%, P < 0.001). Overall median progressive motility as benchmarked by the WHO 2010 criteria was significantly higher in the consecutive sample (43% versus 25%, P < 0.001). Semen analyses of consecutive semen samples collected 30 min (mean) apart in oligozoospemic men should be checked routinely for diagnostic purposes and for managing potential subfertility treatment.

The Sertoli cell: Novel clinical potentiality.

The Sertoli cell is important for endocrine and paracrine control of spermatogenesis. Functions attributed to Sertoli cells are: (1) supportive and trophic functions for the cells of the seminiferous epithelium, (2) transport of mature spermatids towards the lumen of seminiferous tubules, (3) secretion of androgen binding protein, (4) production of substances with endocrine or paracrine action for spermatogenesis control and (5) interaction with intertubular endocrine Leydig cells. Inhibin B and anti-Müllerian hormone (AMH) are glycoproteins belonging to the transforming growth factor β (TGF-β) superfamily; they are produced almost exclusively by the Sertoli cells and have been proposed as direct markers of their function and indirect markers of spermatogenesis. Serum inhibin B and AMH concentrations seem to constitute additional diagnostic parameters in male subfertility as they reflect Sertoli cell function. Stimulated concentrations of serum inhibin B and AMH do not add clinically relevant information in subfertile men compared to basal concentrations of these hormones. Serum inhibin B and AMH concentrations correlate with testicular histology/cytology but are not superior to FSH as predictors of the presence of sperm in testicular sperm extraction (TESE)/fine needle aspiration (FNA) biopsy in men with azoospermia.

Is Varicocelectomy Useful for Subfertile Men with Isolated Teratozoospermia?

To evaluate the effect of microsurgical varicocelectomy on sperm morphology in subfertile men with isolated teratozoospermia and a clinically palpable varicocele. A retrospective review was performed of men with isolated teratozoospermia who had undergone microsurgical varicocelectomy. Semen analyses following varicocelectomy were performed at 2- to 3-month intervals. Responders to the varicocelectomy were defined as those who fulfilled the following criteria in semen analysis: (1) an improvement in percentage of normal forms to ≥4% and (2) normal results for all other semen parameters. A total of 80 patients underwent unilateral varicocelectomy (n = 49) or bilateral varicocelectomy (n = 31) (mean age of 36 ± 0.4 years, range 24-44). After a mean postoperative follow-up period of 6.8 months, the mean percentage of normal sperm forms increased from 0.9% to 3.5% (P < .001), and the mean sperm motility improved from 50.5% to 56.6% (P = .004). The number of responders to the surgery was 16 (20%), and their mean percentage of normal forms increased from 1.3% to 9.1%. The mean preoperative and postoperative semen volume and sperm concentration were not different (P > .05). No differences were observed in age, current smoking status, body mass index, or grade of varicocele between responders and nonresponders. Varicocelectomy might be an option for treating subfertile patients with isolated teratozoospermia and a clinically apparent varicocele. However, patients should be advised that the surgery helps in limited patients only because more patients did not benefit from the surgery.

The safety and efficacy of clomiphene citrate in hypoandrogenic and subfertile men.

Our objective was to evaluate the safety and efficacy of clomiphene citrate (CC) in infertile and hypoandrogenic men through a retrospective study between September 2013 and May 2014. We identified 47 men between 18 and 55 years placed on 50 mg CC every other day. We evaluated the effect of CC on testosterone after 2 weeks, rates of adverse effects and predictors of CC response. Mean baseline testosterone, bioavailable testosterone and estradiol were 246.8 ng dl(-1), 125.5 ng dl(-1) and 20.8 pg dl(-1), respectively. At 2 weeks, mean testosterone, bioavailable testosterone and estradiol increased to 527.6 ng dl(-1), 281.8 ng dl(-1) and 32.0 pg dl(-1) (all P<0.001). Two patients at 2 weeks and one patient at 3 months had a paradoxical decrease in testosterone. Mean total motile count (TMC) and concentration increased from 59.7 million (s.e.m.: 16.5) and 50.7 millions ml(-1) (s.e.m.: 11.1) at baseline to 90.9 million (s.e.m.: 25.9) and 72.5 millions ml(-1) (s.e.m.: 17.5), respectively, at 3 months, although this was nonsignificant (P=0.09, 0.09). No patient on CC experienced a paradoxical decrease in TMC or sperm concentration. On age-adjusted regression analysis, age, BMI, longitudinal testis axis, baseline follicle-stimulating hormone, LH and estradiol did not correlate with improvement in bioavailable testosterone at 2 weeks. CC improves testosterone and may improve semen parameters, although a small percentage of men may not demonstrate improvement in testosterone.

A single-arm pilot study on effects of acupuncture treatment on semen parameters of subfertile Singaporean men

Abstract Objective To evaluate the effect of acupuncture treatment on semen parameters and the proportion of mature and functional sperm as measured by using sperm-hyaluronan binding assay (HBA) in subfertile Singapore men. Methods Singaporean male subjects undergoing subfertility investigation were enrolled prospectively in this pilot study. Each subject received acupuncture treatment at fixed acupoints and two acupuncture sessions per week for 10 weeks. A total of five semen samples were obtained from each subject at 5 weekly intervals up to 20 weeks of the study; at Pre-treatment, Treatment 1 (5th week), Treatment 2 (10th week), Post-treatment 1 (15th week) and Post-treatment 2 (20th week). Semen analysis was performed to assess semen parameters and HBA score (percentage of motile sperm bound to hyaluronan) to determine the proportion of mature and functional sperm. Results Mean HBA scores improved significantly from 12.0% at Pre-treatment, to 18.4% (P = 0.03) at 5th week and was 16.8% (P = 0.07) at 10th week of acupuncture treatment. After completion of acupuncture treatment, HBA scores continued to increase significantly to 25.8% (P Conclusions HBA scores increased significantly during the first 5 weeks of acupuncture treatment (Treatment 1) and 5 weeks after the completion of treatment (Post-treatment 1). Hence acupuncture treatment was effective in increasing the proportion of mature and functional sperm but had no effect on other semen parameters.

No association between body mass index and sperm DNA integrity.

Is overweight associated with impaired sperm DNA integrity? High body mass index (BMI) is not associated with impaired sperm DNA integrity as assessed by the DNA Fragmentation Index (DFI). Previous studies, based on fewer subjects and including mainly subfertile men, have shown conflicting results regarding the influence of overweight and obesity on sperm DNA integrity. This cross-sectional study was based on semen samples from 1503 men from the general population. We included two cohorts (cohort A and B) of military recruits (n = 275, n = 304, respectively), one group (cohort C) of fertile men and men without known fertility problems (n = 724), and one group (cohort D) of men between 19 and 40 years without known fertility problems (n = 200). In all cohorts, data were available on BMI, DFI as measured by the sperm chromatin structure assay (SCSA), standard semen characteristics, and potential confounders (age, abstinence time, smoking habits). The subjects were categorized according to BMI into four groups: underweight (<18.5 kg/m(2)), normal weight (18.5-24.9 kg/m(2)), overweight (25.0-29.9 kg/m(2)) and obese (≥30.0 kg/m(2)). Using a linear regression model, the inter-group differences in DFI were calculated. Furthermore with the normal-weight group as the reference, the odds ratios (ORs) for DFI > 20% and DFI > 30%, were calculated for the other groups. Calculations were made for the material as a whole and after exclusion of cohort C which included proven fertile men. We found that normal-weight men had significantly higher DFI than overweight men, with a mean difference of 1.13% (95% CI: 1.05-1.22%); P = 0.001). Overweight men had a reduced risk of having DFI ≥ 20% and DFI ≥ 30%, compared with normal-weight men; adjusted odds ratio (OR) = 0.61 (95% CI: 0.42-0.88; P < 0.01) and adjusted OR = 0.48 (95% CI: 0.28-0.84; P < 0.01), respectively. When excluding cohort C, the statistical significance was lost. Regarding standard semen parameters, we found that obese men had a higher percentage of progressive motile spermatozoa than normal-weight men; mean difference 1.15% (95% CI: 1.02-1.30%, P < 0.05) but the significance was lost when excluding cohort C. All other standard semen parameters were unaffected by BMI. A main limitation might be the cross-sectional nature of the data. Furthermore our study included a significant proportion of men with proven fertility (75% of cohort C, n = 550), and could therefore be biased toward fertility. Our study indicates that overweight per se is not associated with a higher level of sperm DNA damage. This research has been given grants from the following: EU 5th and 7th framework program (Inuendo and Clear projects, [Contracts no. QLK4-CT-2001-00202 and FP7-ENV-2008-1-226217)]), the Swedish Research Council (Grants No. 2007-2590, 521-2004-6072 and 521-2002-3907); the Swedish Governmental Funding for Clinical Research, Skåne county council's research and development foundation, MAS Funds, University Hospital MAS Foundation in Malmö, Crafoordska Fund, Ove Tulefjords Fund, Foundation for Urological Research, Fundacion Federico SA, and Gunnar Nilssons Cancer Fund. The authors declare that there are no conflicts of interest.

Methylation analysis of histone H4K12ac-associated promoters in sperm of healthy donors and subfertile patients.

BackgroundHistone to protamine exchange and the hyperacetylation of the remaining histones are hallmarks of spermiogenesis. Acetylation of histone H4 at lysine 12 (H4K12ac) was observed prior to full decondensation of sperm chromatin after fertilization suggesting an important role for the regulation of gene expression in early embryogenesis. Similarly, DNA methylation may contribute to gene silencing of several developmentally important genes. Following the identification of H4K12ac-binding promoters in sperm of fertile and subfertile patients, we aimed to investigate whether the depletion of histone-binding is associated with aberrant DNA methylation in sperm of subfertile men. Furthermore, we monitored the transmission of H4K12ac, 5-methylcytosine (5mC) and 5-hydroxymethylcytosine (5hmC) from the paternal chromatin to the embryo applying mouse in vitro fertilization and immunofluorescence.ResultsChromatin immunoprecipitation (ChIP) with anti-H4K12ac antibody was performed with chromatin isolated from spermatozoa of subfertile patients with impaired sperm chromatin condensation assessed by aniline blue staining. Fertile donors were used as control. DNA methylation analysis of selected H4K12ac-interacting promoters in spermatozoa was performed by pyrosequencing.Depletion of binding sites for H4K12ac was observed within the following developmentally important promoters: AFF4, EP300, LRP5, RUVBL1, USP9X, NCOA6, NSD1, and POU2F1. We found 5% to 10% hypomethylation within CpG islands of selected promoters in the sperm of fertile donors, and it was not significantly altered in the subfertile group. Our results demonstrate that the H4K12ac depletion in selected developmentally important promoters of subfertile patients was not accompanied by a change of DNA methylation.Using a murine model, immunofluorescence revealed that H4K12ac co-localize with 5mC in the sperm nucleus. During fertilization, when the pronuclei are formed, the paternal pronucleus exhibits a strong acetylation signal on H4K12, while in the maternal pronucleus, there is a permanent increase of H4K12ac until pronuclei fusion. Simultaneously, there is an increase of the 5hmC signal and a decrease of the 5mC signal.ConclusionsWe suggest that aberrant histone acetylation within developmentally important gene promoters in subfertile men, but not DNA methylation, may reflect insufficient sperm chromatin compaction affecting the transfer of epigenetic marks to the oocyte.Electronic supplementary materialThe online version of this article (doi:10.1186/s13148-015-0058-4) contains supplementary material, which is available to authorized users.

Meiotic and sperm aneuploidy studies in three carriers of Robertsonian translocations and small supernumerary marker chromosomes

To study the meiotic behaviour of one carrier of a small supernumerary marker chromosome (sSMC): 47,XY,+mar; one carrier of a Robertsonian translocation (ROB): 45,XY,rob(13;21) (q10;q10); and one carrier of both a sSMC and a ROB: 46,XY,rob(13;21) (q11.1;q11.1),+mar. Case-control study. University-affiliated research center and hospital. Subfertile men with ROB and sSMC. None. The chromosomal origin of the sSMC was assessed by multiplex fluorescence in situ hybridization. The segregation of the ROB and sSMC in sperm and possible interchromosomal effects were examined by fluorescence in situ hybridization. Synapsis, meiotic recombination, and meiotic inactivation were investigated in ejaculate spermatocytes of the 47,XY,+mar and 45,XY,rob(13;21) carriers using immunostaining. In the 47,XY,+mar and 46,XY,rob(13;21),+mar carriers, the sSMC was found in 13.5% and 11.5 % of sperm, respectively. Analysis of meiotic segregation of chromosome 13 and 21 showed that 91.2% of sperm were normal/balanced in the 46,XY,rob(13;21),+mar case, whereas 88.4% of sperm were normal/balanced in the 45,XY,rob(13;21) case. Interchromosomal effects involving the sex chromosomes were found in both sSMC carriers. Both 47,XY,+mar and 45,XY,rob(13;21) carriers showed decreased global recombination, impaired synapsis, and an association of abnormal chromosomes with the XY body. Carriers of marker chromosomes produce sperm with markers at frequencies lower than theoretically expected. Carriers of ROB and sSMC showed decreased recombination, impaired synapsis, and association of abnormal chromosomes with the XY body, which may contribute to an interchromosomal effect. Using immunofluorescence techniques to analyze ejaculate-derived spermatocytes from subfertile men provides a novel technique for examining meiosis without the need for a testicular biopsy.

Serum miR-155 as a potential biomarker of male fertility.

Are serum levels of micro-RNAs miR-155 and miR-146a associated with male fertility, low-grade systemic inflammation (LGSI) and androgens? miR-155 was associated with male subfertility independent of LGSI or androgens while miR-146a was only weakly associated with subfertility and LGSI. Male subfertility has been associated with LGSI as well as with androgen deficiency. miR-155 and miR-146a are central regulators of inflammation and their level in cells and in the serum has been associated with several inflammatory conditions. In this case-control study, two independent groups of 60 subjects each (exploratory and confirmatory cohort) were randomly selected from an ongoing study on subfertile men (in total: hypogonadal; n = 40, eugonadal; n = 40 and control group n = 39) at a University Hospital Reproductive Medicine Centre. Individuals were matched for age. Total RNA was isolated from cell-free serum. As internal control a synthetic miRNA, UniSp6, was added to each sample prior to extraction. miRNA expression levels were measured by real-time RT-PCR and presented as fold difference (arbitrary units, U) from control. Sera from these individuals had been previously analyzed for hormone and cytokine levels. Serum levels of miR-155 were associated with levels of miR-146a (P < 0.0001), but only miR-146a was associated with inflammatory markers. miR-155 was strongly associated with subfertility (for subfertile group 1.88 U, 95% confidence interval (CI) 1.6-2.1 U versus 1.15, 95% CI 1.0-1.2 U in controls; P = 0.001). Receiver operating characteristic curve analysis indicated that miR-155 but not miR-146a can be used as a marker of subfertility. MiR-155 with a cutoff value of 1.77 had 47% sensitivity and 95% specificity for identifying subfertility and a positive predictive value (PPV) and negative predictive value (NPV) of 95 and 47%, respectively. When used in combination with FSH, sensitivity and specificity were 80 and 100%, respectively, while PPV and NPV were 100 and 71%, respectively, those values being higher than for the FSH alone. Repeating the results obtained in the exploratory cohort in an independent confirmatory cohort reduced the risk of a chance finding. Although the results from the exploratory cohort were confirmed in the confirmatory cohort, studies from other centers are needed to establish the role of miR-155 as a new biomarker of male fertility. Furthermore, the role of this marker in distinguishing between different groups of male subfertility is to be elucidated. Association of the inflammatory miRNA miR-155 with male fertility contributes to our understanding of the pathophysiology of subfertility and suggests a novel biomarker. Serum miR-155 in combination with FSH has higher diagnostic specificity and sensitivity compared with FSH alone. This work was supported by grants from Swedish Governmental Grant (ALF), Skane county council research and development foundation, Skane University Hospital Fonds and by the EU and Greek funds under the action 'Education and lifelong learning' program THALIS-FAT-VESSEL (No 379527). The authors have no competing interests to disclose.

Conventional semen parameters and DNA fragmentation in relation to fertility status in a Greek population

ObjectivesOur study aimed to investigate the possible correlations between conventional seminal parameters and DNA fragmentation in specific groups of Greek men, selected in relation to their fertility history and to verify the validity of the recent WHO reference values for the basic semen analysis in this population. Study designA total of 770 subject data were evaluated in three distinct groups: fertile men with children naturally conceived within one year of unprotected intercourse (n=78), subfertile men, having achieved pregnancies either naturally or by Assisted Reproduction Techniques, not resulting in live births (n=153) and infertile men, failing to produce either pregnancies or children (n=539). Semen volume, sperm concentration, total count, rapid and total progressive motility and morphology were evaluated following the World Health Organization (2010) methods. DNA fragmentation was assessed by the Sperm Chromatin Dispersion assay. The 5th percentile, as well as the 95% confidence intervals (CI) for each parameter, were calculated by the method of bootstrapping. Statistical correlations between the examined parameters were sought using the Spearman R test (p<0.05). ResultsA significant inverse correlation was established between DNA fragmentation and sperm concentration, total count, progressive motility (rapid and total) and normal morphology in subfertile and infertile men (p<0.05). No statistically significant correlations were remarked between the conventional semen parameters and the levels of DNA fragmentation in our group of fertile men. Concordance was established between the reference limits issued by the WHO 2010 for the basic semen parameters and semen quality of fertile men in the studied population. ConclusionsThe variability of correlations established between DNA fragmentation and the conventional seminal profile in relation to fertility status indicates that they are independent attributes of semen quality, justifying the assessment of both during a comprehensive evaluation of male infertility. Moreover, the WHO 2010 reference limits were found adequately descriptive of seminal normality in Greek men.

Seminal parameters of chronic male genital inflammation are associated with disturbed sperm DNA integrity.

Definition of chronic male genital tract inflammation and its impact on male infertility is still a matter of debate. In particular, DNA integrity has been reported to be disturbed in subfertile men. Thus, the aim of this study was to investigate an association of DNA integrity to altered standard semen parameters as well as inflammatory parameters such as peroxidase-positive cells, macrophages and seminal interleukin-6 concentration. Macrophages were detected by CD18/HLA-Dr staining, and DNA integrity was analysed by acridine orange staining using flow cytometry. Interleukin-6 was detected by ELISA. Normal DNA integrity showed a significant correlation to sperm number and progressive motility. Moreover, a significant inverse correlation of DNA integrity to Interleukin-6 and macrophages could be demonstrated. Further on, seminal interleukin-6 also significantly correlated to macrophages. No association has been observed between the number of peroxidase-positive cells and normal DNA integrity. As disturbed DNA integrity has been reported to negatively influence spermatozoon-egg interaction and even fertilisation rates following ICSI, and as early miscarriages have been associated with sperm DNA damage, it should be screened very carefully for male genital tract inflammations in couples undergoing infertility treatment. Measuring Interleukin-6 seems superior to assessment of the number of leucocytes alone and additional assessment of DNA integrity into the diagnostic work-up should be considered.

Treatment of infertile and subfertile men with varicocelle by minimally invasive surgery

Treatment of infertile and subfertile men with varicocelle by minimally invasive surgery

Translocator Protein in Mitochondrial Cholesterol Transport and the Pharmacology of Steroidogenesis

Steroidogenesis begins with the transfer of cholesterol from intracellular stores into mitochondria through a complex formed of cytosolic proteins, such as the steroidogenesis acute regulatory protein and 14-3-3 adaptor, and outer mitochondrial membrane proteins Translocator Protein (TSPO) and Voltage Dependent Anion Channel (VDAC). TSPO is an evolutionary conserved 18-kDa high affinity drug- and cholesterol- binding protein expressed in high levels in steroid synthesizing cells. Aberrant expression of TSPO has been linked to cancer, neurodegeneration, neuropsychiatric disorders and primary hypogonadism. TSPO drug ligands have been proposed as therapeutic and monitoring agents for these diseases. In the brain, steroids are local regulators of neural development and excitability. Changes in neurosteroid levels are linked to neuropsychiatric and neurological disorders such as depression, anxiety and neurodegeneration. Local administration of neurosteroids is unfeasible, and treatment of patients with large amounts of neuroactive steroids unsafe. In the testis, reduced serum testosterone (T) is common among subfertile and infertile young men as well as aging men and is often associated with depression, metabolic syndrome, and reduced sexual function. Although T-replacement therapy has been the treatment of choice, there are numerous side-effects. Thus, there is a clear need for developing repair therapies that restore the brain's and testis’ abilities to make steroids. In vitro and in vivo studies, using animal models of disease, demonstrated that TSPO drug ligands increased neurosteroid production in neuropsychiatric disorders and T formation in hypogonadism. Moreover, peptides conjugated to 14-3-3e site of interaction with VDAC1 blocked 14-3-3e-VDAC1 and increased VDAC1-TSPO interactions in testis inducing T formation. In contrast, in constitutively steroid producing Leydig and adrenal cell tumors blocking TSPO function inhibits excessive steroid synthesis. TSPO and VDAC were identified as valid drug targets that control steroid formation both in vitro and in vivo.

Antioxidants for male subfertility.

Antioxidants for male subfertility.

Body mass index, waist-to-hip ratio, waist circumference and waist-to-height ratio cannot predict male semen quality: a report of 1231 subfertile Chinese men.

There were controversial results between obesity-associated markers and semen quality. In this study, we investigated the correlations between age, obesity-associated markers including body mass index (BMI), waist-to-hip ratio (WHR), waist-to-height ratio (WHtR) and waist circumference (WC), the combination of age and obesity-associated markers, semen parameters and serum reproductive hormone levels in 1231 subfertile men. The results showed that BMI, WC, WHR and WHtR were positively related to age, and there were also positive relations between BMI, WHR, WC and WHtR and between sperm concentration (SC), total sperm count (TSC), progressive motility (PR), sperm motility and per cent of normal sperm morphology (NSM). However, age, each of obesity-associated markers and the combination of obesity-associated markers and age were unrelated to any of semen parameters including total normal-progressively motile sperm count (TNPMS). Age, BMI, WHR, WC and WHtR were negatively related to serum testosterone and SHBG levels. However, only serum LH and FSH levels were negatively related to sperm concentration, NSM and sperm motility. In a conclusion, although age and obesity have significant impacts on reproductive hormones such as testosterone, SHBG and oestradiol, semen parameters related to FSH and LH could not be influenced, indicating that obesity-associated markers could not predict male semen quality.