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Male reproductive problems may derive from many reasons including the environmental toxicants which may either intaken by occupational exposure, nutrition or bad air quality. The increased exposure to these substances due to rapid industrial development and technology has raised the questions: Is there a relationship between sperm parameters and these substances, and if so, in what extent? Results of studies on the subject reported conflicting results, many of which were not investigated in the seminal plasma. The aim of the current study was to evaluate the relationship between 23 metals and trace elements in human seminal plasma and semen parameters many of which were not investigated before. Levels of 23 metals in human seminal plasma were assessed by inductively coupled plasma mass spectrometry (ICP‐MS). We examined the differences between subjects with normal ejaculate (normozoospermia) and pathologic ejaculate (with at least one abnormal semen parameter) according to the WHO criteria. The only significant difference was detected for Se while the other element's difference was not statistically significant. Se was statistically significantly increased in normal semen group suggesting the positive effect of this element on semen parameters. [ABSTRACT FROM AUTHOR]

The incidence of sub‐fertility is higher in crossbred bulls compared to zebu bulls. In the present study, we analysed the metabolomic profile of seminal plasma from crossbred and zebu bulls and uncovered differentially expressed metabolites between these two breeds. Using a high‐throughput LC‐MS/MS‐based approach, we identified 990 and 1,002 metabolites in crossbred and zebu bull seminal plasma respectively. After excluding the exogenous metabolites, we found that 50 and 68 putative metabolites were unique to crossbred and zebu bull seminal plasma, respectively, whilst 87 metabolites were common to both. After data normalisation, 63 metabolites were found to be dysregulated between crossbred and zebu bull seminal plasma. Observed pathways included Linoleic acid metabolism (observed metabolite was phosphatidylcholine) in crossbred bull seminal plasma whereas inositol phosphate metabolism (observed metabolites were phosphatidylinositol‐3,4,5‐trisphosphate/inositol 1,3,4,5,6‐pentakisphosphate/myo‐inositol hexakisphosphate) was observed in zebu bull seminal plasma. Abundance of Tetradecanoyl‐CoA was significantly higher, whilst abundance of Taurine was significantly lower in crossbred bull seminal plasma. In conclusion, the present study established the seminal plasma metabolomic profile in crossbred and zebu bulls and suggest that increased lipid peroxidation coupled with low concentrations of antioxidants in seminal plasma might be associated with high incidence of sub‐fertility in crossbred bulls. [ABSTRACT FROM AUTHOR]

Summary: The present study aimed to compensate dilution effect using additional seminal plasma (SP) in conventional (80 million (M) spermatozoa/ml) dose and low spermatozoa/dose (8M spermatozoa/ml). We also attempted to confirm whether removal of SP before the extension of ejaculates affects post‐thaw sperm quality of buffalo semen. For this, semen ejaculates (N = 15) were divided into four groups: control (CON), removal of SP by centrifugation (NSP), resuspension of the centrifuged semen pellet into SP (CEN) and extra supplementation of SP (ESP). All groups were diluted into two different semen doses to 20 and 2M spermatozoa/0.25 ml using tris egg yolk extender and subsequently cryopreserved. We found that neither addition nor removal of SP affected sperm motility, kinematics, longevity, mitochondrial superoxide production and high mitochondrial membrane potential (MMP). Further, the addition or removal of SP was not able to compensate dilution effect in 2M groups resulting in a significantly (p <.05) reduction in sperm motility, kinematics, sperm longevity, membrane integrity, MMP, and an increase production of mitochondrial superoxide. In conclusion, it appears that role of SP in the sperm cryopreservation process is insignificant. [ABSTRACT FROM AUTHOR]

The aim of the study was to investigate the efficiency of ram seminal plasma and fetal calf serum on freezing of buck semen. Twenty ejaculates were collected using an electro‐ejaculator and split into six groups. While FCS additive was not used in A1, A2 and A3 groups, 10% FCS was added to B1, B2 and B3 groups. These groups were then edited according to whether the buck or ram SP was involved. The design of the groups was done as follows: Group A1 (control 1), group A2 without buck SP, group A3 containing ram SP instead of buck SP. Groups B1 (control 2), B2 and B3 were the FCS added forms of these groups. Progressive sperm motility percentages in Group A1 and Group B2 were found to be higher when compared to the lowest Group B3. There were no significant differences between the groups in neither the levels of reactive oxygen species nor the enzyme and glutathione activities. In conclusion, the lack of statistical difference between the groups suggested that despite the supplements used but only when the buck spermatozoa structure was healthy, the cell could preserve acrosome, DNA and the integrity of membrane. [ABSTRACT FROM AUTHOR]

Oxidative stress (OS) has been shown to have a key role in male infertility. Recently, a new measurement method has been developed to measure the overall oxidation–reduction potential (ORP) in a semen sample known as the MiOXSYS system. The aim of this study was to investigate the correlation of sperm parameters with oxidative stress levels determined by ORP and to evaluate whether the current limit is able to distinguish abnormal sperm parameters from normal ones in Turkish population. Semen samples of 121 patients who applied for infertility investigation were divided into two groups as (OS +; n:39) and (OS −; n:82). Semen parameters were compared between groups. Sperm concentration, total motility and progressive motility were found to be significantly lower in OS (+) patients compared to those OS (−), while immotile sperm count was significantly higher in OS (+) patients. Oxidative stress determined by MiOXSYS system was found to be related to reduced sperm parameters in Turkish population, which may be used as an indicator of poor sperm parameters and a support to routine semen analysis. In addition, recommended reference value was found to be reliable in distinguishing normal from impaired semen parameters. [ABSTRACT FROM AUTHOR]

Prostate‐specific membrane antigen (PSMA) is a transmembrane glycoprotein with glutamate carboxypeptidase activity. However, its precise function in the prostate, prostasomes and seminal plasma with regard to male fertility remains unknown. This study was conducted to investigate the seminal plasma PSMA levels in fertile men and patients with oligoasthenoteratozoospermia (OAT) and to analyse its association with sperm parameters. Twenty fertile men and twenty patients admitted at the urology clinic of our institution with the diagnosis of OAT were included in the study. Following semen analysis, seminal plasma was isolated from semen ejaculates. PSMA concentrations in the seminal plasma were determined by ELISA. The correlations between seminal PSMA concentrations and semen parameters were statistically analysed. Seminal plasma PSMA concentration was significantly lower in OAT patients compared to fertile controls (p <.01). In fertile men, PSMA concentration was significantly correlated with the sperm concentration (r = −.481, p <.05), whereas in the patient group no statistically significant correlation was found between the sperm parameters and seminal PSMA level. This is the first study in the literature to investigate PSMA levels in the seminal plasma from infertile men. Decreased levels of seminal plasma PSMA might suggest a role for compromised prostasome function in the pathogenesis of OAT syndrome. [ABSTRACT FROM AUTHOR]

Varicocele is one of the major causes of male infertility and has a negative impact on spermatogenesis. The conventional semen analysis does not reveal the underlying subcellular mechanisms associated with defects in spermatozoa. Proteomics and bioinformatics analysis can be used to identify the molecular aetiologies associated with poor semen quality in varicocele patients. Mitochondrial dysfunction has been identified as the main factor affecting normal physiological functions of spermatozoa. This article discusses the proteomic studies of spermatozoa and seminal plasma in varicocele patients. Proteomics can identify potential spermatozoa and seminal plasma biomarkers in varicocele‐mediated male infertility. In future, these protein biomarkers can be useful in the development of noninvasive diagnostic and therapeutic strategies for varicocele patients. [ABSTRACT FROM AUTHOR]

Seminal plasma is intimately connected to sperm physiology and particularly in South American Camelids, has demonstrated to be involved in multiple physiological reproductive events. Different percentages of seminal plasma (0%, 10% and 50%) were added to thawed llama semen samples with the objective of evaluating the interaction with cryopreserved sperm over time (0, 1.5 and 3 hr at 37°C). A total of 20 ejaculates from five adult llama males (n = 5; r = 4) were evaluated. A significant decrease in sperm motility, membrane function and live sperm was observed in all thawed samples (0%, 10% and 50%) at 0 hr when compared to raw semen. Neither morphology nor chromatin condensation was altered in all thawed samples (p >.05), but a significant increase in the percentage of spermatozoa with fragmented DNA was observed after thawing all samples versus raw semen. When evaluating thawed samples over time, a significant decrease of motility and membrane function was observed, while the percentages of total live sperm were preserved over the 3 hr of incubation in all final concentrations evaluated. To conclude, the addition of 10% or 50% of seminal plasma was incapable of preserving motility or membrane function of frozen‐thawed llama sperm during 3 hr of incubation. [ABSTRACT FROM AUTHOR]

There is growing evidence that vitamin B6 has a valuable contribution in maintaining normal sperm parameters; however, this contribution has not yet well‐identified. Here, we aimed to measure the level of seminal plasma vitamin B6 in men with asthenozoospermia compared to men with normal sperm motility. Ninety‐seven human males with asthenozoospermia and eighty‐eight human males with normal sperm motility (control) were recruited in this study. Collected semen samples were assessed for sperm motility, sperm count and semen volume. Liquid chromatography with tandem mass spectrometry was used to measure seminal plasma vitamin B6 concentrations. A highly significant difference (p <.0001) in concentrations of seminal plasma vitamin B6 was found between asthenozoospermic and control groups. Besides, no statistical correlations were found between seminal plasma vitamin B6 level and sperm motility, sperm count, semen volume and men age in both tested groups. In conclusion, men with asthenozoospermia have lower seminal plasma vitamin B6 level compared to men with normal sperm motility. Also, seminal plasma vitamin B6 was found not to be correlated with sperm motility and count, semen volume and men age in both tested groups. These results may provide new contribution in the management of male infertility. [ABSTRACT FROM AUTHOR]

The objective of this study was to determine the optimum concentrations of rainbow trout seminal plasma (RTS) supplemented extenders for goat semen quality at post‐thaw and after incubation. Five sexually mature Saanen goat (Capra aegagrus hircus) were used for semen collection. Pooled semen was diluted with soybean lecithin‐based extender without RTS (control) or supplemented with different concentrations of RTS (1%, 2%, 4% or 8%), at a final concentration of 150 × 106 spermatozoon/ml. Sperm motility, plasma membrane functional integrity (HOST), damaged acrosome (PSA‐FITC), mitochondrial activity (rhodamine123) and DNA integrity (TUNEL) were evaluated. Spermatological parameters were evaluated at post‐thaw and after 6 hr incubation. RTS8 group preserved sperm motility, acrosomal integrity, plasma membrane functional integrity and mitochondrial function better than the control group (p <.05). The study demonstrated that RTS supplemented lecithin‐based extenders have useful effects on goat spermatozoa. In addition, the results of the current study represented the positive effect of using 8% RTS supplemented extender. [ABSTRACT FROM AUTHOR]

MicroRNAs (miRNAs) have recently received a significant amount of attention due to their remarkable influence on post‐transcriptional gene regulation. In this study, we aim to provide a catalogue of miRNAs present in spermatozoa, seminal plasma and testicular tissue. Expression profiles of miRNA in spermatozoa and seminal plasma of 16 proven fertile men and testicular tissue of eight men with morphologically and/or histologically confirmed obstructive azoospermia were determined by microarray and RT‐qPCR in combination with bioinformatics analyses. A total of 123, 156 and 133 miRNAs were consistently detected in spermatozoa, seminal plasma and testicular tissue respectively. Sixty‐four miRNAs were shared across all sample types. Based on miRNAs expression level present in each group, correlation analysis showed moderate‐to‐strong correlations within the spermatozoa and seminal plasma samples and a wider range of correlations within the testicular tissue samples. The target genes of known miRNAs appeared to be involved in a wide range of biological processes related to reproduction, development and differentiation of germ cells. Our results suggest that there is a certain similarity between spermatozoa and seminal plasma for the relative miRNA expression changes with respect to testicular tissue and provide an overview of the miRNAs present in each sample type. [ABSTRACT FROM AUTHOR]

Cell‐free seminal mRNA (cfs‐mRNA) exists in the human ejaculate that is proposed as a potential noninvasive procedure to prognosis pathophysiological conditions. This study applied cfs‐mRNA of ESX1, ZMYND15 and its target haploid genes (TNP1 and PRM1) to identify the presence of spermatozoa in men with azoospermia. This study included 35 semen samples from 16 normozoospermic and 19 nonobstructive azoospermic individuals. Expression levels of target genes were determined by real‐time reverse transcription–polymerase chain reaction using ΔΔCt method. The expression level of these genes (ZMYND15, TNP1 and PRM1) was significantly decreased in semen samples of nonobstructive azoospermia compared to normozoospermia. Similarly, the expression level of TNP1 and PRM1 was significantly decreased in the sample with negative sperm (SR−) versus positive sperm retrieval (SR+). The expression level of these genes may have the potential for prediction of successful sperm retrieval with high sensitivity and specificity according to the receiver operating characteristics (ROC) curve analysis. [ABSTRACT FROM AUTHOR]

Abstract: Growth hormone (GH) and insulin‐like growth factor 1 (IGF‐1) have been proposed to play a pivotal role in male infertility due to their anabolic effects. The aim of this study was to investigate possible associations between seminal plasma levels of GH and IGF‐1 and sperm parameters. Fifty men participated in this study. Semen analysis was performed, while cell‐free seminal plasma was collected following sperm centrifugation. Seminal plasma concentrations of IGF‐1 and GH were determined by enzyme‐linked immunosorbent assay (ELISA). Due to the presence of asthenozoospermia in all participants who presented with abnormal sperm parameters, the participants were further subdivided into normal (group A), asthenozoospermic (group B) and asthenozoospermic plus at least one additional abnormal parameter (group C). A marginally nonsignificant statistical difference (p = 0.063) was revealed between the GH levels corresponding to the asthenozoospermic and the normal group with the latter presenting with higher GH levels. A statistically significant positive correlation (p < 0.05) was noted between levels of GH and IGF‐1 in group C. The above relationship has also been observed in men with low sperm concentration, vitality, volume and abnormal morphology. These novel findings require further investigation in order for the biological significance of those associations to be clarified. [ABSTRACT FROM AUTHOR]

To achieve the targeted artificial insemination coverage with the current rate of semen production, without affecting the conception rate, it needs to reduce the number of spermatozoa per insemination dose in India as per international practice. Therefore, this study was planned to perform different levels of semen dilution, compare in vitro post-thaw semen quality and develop a modified low-dose semen packaging method in French mini straw to minimise semen dilution effect. Sixteen ejaculates were collected from Karan Fries bulls (n = 4). The mean percentage postthaw motility, viability, membrane integrity, acrosome integrity, lipid peroxidation and capacitation status were estimated as post-thaw sperm function assays in semen sample diluted to 20, 15, 10 and 5 million spermatozoa per 0.25 ml and filled in the French mini straw by conventional packaging. No significant (p > .05) difference in post-thaw sperm quality was observed between 15 and 20 million doses; however, below 15 million sperm quality get reduced. There was no significant difference in post-thaw semen quality traits between 20 million conventional packaging and 5 million spermatozoa/dose in modified packaging. In conclusions, the modified packaging is a very effective method for low-dose cryopreservation with acceptable post-thaw semen quality. [ABSTRACT FROM AUTHOR]

Summary: The aim of this study was to determine the effects of both the removal of seminal plasma (SP) and the pre‐freezing addition of seminal plasma collected during the breeding or nonbreeding season on goat sperm survival after thawing. Semen samples were pooled. One aliquot of pooled semen was used as a control group. Four aliquots were then centrifuged, and the SP was removed in Group I, pipetted but not removed in Group II, removed and then pooled for animals collected in the breeding season in Group III and removed and pooled for animals collected in the nonbreeding season in Group IV. Group samples were frozen and then were assessed for rates of sperm motility, plasma membrane functional integrity hypo‐osmotic swelling test (HOST), defective acrosomes (FITC‐PSA), DNA fragmentation (TUNEL) and mitochondrial membrane damage (Rhodamine 123). The results showed that pre‐freezing addition of SP collected in breeding season maintained post‐thaw sperm characteristics at 0 hr better than SP removal group, but removing seminal plasma showed positive effects on spermatozoa, as incubation time increased to 5 hr. In conclusion, the pre‐freezing addition of seminal plasma did not maintain post‐thaw goat sperm characteristics as successfully as in the groups with seminal plasma removed after an incubation period. [ABSTRACT FROM AUTHOR]

The mechanism of asthenozoospermia remains unclear. The knowledge of the metabolism of fatty acids in seminal plasma is important and meaningful for the pathological study of asthenozoospermia. We present an optimised assay of extraction and derivatisation followed by GC/ MS to analyse metabolites, especially fatty acids, in seminal plasma from healthy and asthenozoospermic men. Eighty-nine peaks including 17 kinds of fatty acids were analysed and identified in the chromatogram. The GC/ MS data were analysed using t test, fold change and partial least squares discriminant analysis to explore the potential biomarkers of asthenozoospermia. Seven metabolites in asthenozoospermic group were found to be significantly different from those in the normal group (with p < .05, fold change >1.2 and variable importance for projection >1). Of which, high levels of oleic acid and palmitic acid in seminal plasma from asthenozoospermic men may indicate a membrane metabolism disorder in spermatozoa and the lack of valine in the asthenozoospermic group may contribute to poor sperm motility. The results may facilitate the understanding of the role of fatty acids and amino acids in asthenozoospermia and provide solid foundation for further pathological study of asthenozoospermia. [ABSTRACT FROM AUTHOR]

The aims of this study were to determine concentrations of total homocysteine, cysteine, cysteinylglycine and glutathione in spermatozoa, seminal fluid and blood plasma and to analyse their relationships with sperm parameters. For this reason, a new highly effective method of spermatozoa lysis was developed, using methanol, freezing and subsequent thawing in ultrasonic bath. An HPLC- FD assay was conducted on thiols concentrations in lysed spermatozoa, seminal fluid and blood plasma. Concentrations of thiols in spermatozoa were significantly lower in men with normozoospermia than in samples with pathological semen parameters. Statistical analysis found significant correlations between thiol concentrations in spermatozoa and semen parameters, while the same analysis with thiol concentrations in seminal fluid was substantially less powerful. Only cysteinylglycine concentrations in seminal fluid significantly correlated with pathological semen parameters. No significant differences or correlations were found with blood plasma concentrations. [ABSTRACT FROM AUTHOR]

The effects of seminal plasma ( SP), derived from autologous, homologous and heterologous species (stallion, boar and dog) on chilled Asian elephant sperm quality, were determined. Semen was collected from eight males and samples with ≥30% motile spermatozoa were used in the study. Semen was diluted with Tris-glucose-egg yolk extender, supplemented with different SP types and preserved at 4°C for 48 hr. Experiment 1 ( n = 31), showed that the presence of SP (autologous) helped to preserve sperm quality in terms of sperm motility and acrosome integrity ( p < .05). Homologous SP did not result in better sperm quality than autologous SP. Heterologous SP from stallion provided higher sperm motility and velocities compared to autologous SP ( p < .05). Experiment 2 ( n = 14) determined the effect of different SP from four stallions. All stallion SP gave higher ( p < .05) results for motile spermatozoa and sperm velocities than autologous SP. In conclusion, the presence of SP helps preserve Asian elephant sperm quality and stallion SP supports the motility of Asian elephant spermatozoa during cold storage. [ABSTRACT FROM AUTHOR]

Seminal plasma ( SP) of South American Camelids could interfere with the interaction of spermatozoa with the extenders; therefore it becomes necessary to improve semen management using enzymatic treatment. Our objective was to compare two cooling protocols for llama semen. Twelve ejaculates were incubated in 0.1% collagenase and then were divided into two aliquots. One was extended in lactose and egg yolk ( LEY) (Protocol A: collagenase and SP present). The other aliquot was centrifuged, and the pellet was resuspended in LEY (Protocol B: collagenase and SP absent). Both samples were maintained at 5°C during 24 hr. Routine and DNA evaluations were carried out in raw and cooled semen. Both cooling protocols maintained sperm viability, membrane function and DNA fragmentation, with Protocol A showing a significantly lowered total and progressive motility ( p < .05) and Protocol B showing a significant increase in chromatin decondensation ( p < .05). Protocol A avoids centrifugation, reducing processing times and making application in the field simpler. However, as neither protocol showed a significant superiority over the other, studies should be carried out in vivo to evaluate the effect on pregnancy rates of the presence of collagenase and SP in semen samples prior to either cooling or freeze-thawing. [ABSTRACT FROM AUTHOR]

The objective of this study was to assess the impact of seminal clusterin level on spermatogenesis in infertile men. This study included 89 men who visited our clinic due to infertility, consisting of 28, 33, and 28 diagnosed with normospermia, oligozoospermia and nonobstructive azoospermia (NOA) respectively. The seminal clusterin concentrations measured by enzyme-linked immunosorbent assay were 47.9, 28.2 and 18.4 ng ml−1 in men with normospermia, oligozoospermia and NOA, respectively, with significant differences among these three groups ( P < 0.01). Microdissection testicular sperm extraction (MD-TESE) was performed in the 28 men with NOA, and spermatozoon was successfully retrieved from 9. There was a significant correlation between seminal clusterin level and testicular clusterin protein expression evaluated by immunohistochemical staining in these men with NOA ( P = 0.026). Of several parameters available before MD-TESE, the univariate analysis identified serum follicle-stimulating hormone (FSH) level <10 IU ml−1 and seminal clusterin level ≥18 ng ml−1 as significant predictors of sperm retrieval, and of these, only serum FSH level <10 IU ml−1 was shown to be independently associated with sperm retrieval in the multivariate analysis. Accordingly, it might be worthy to further evaluate the significance of seminal clusterin level as a biomarker for the assessment of spermatogenic status in infertile men. [ABSTRACT FROM AUTHOR]

Fatty acids are important in male reproductive function because they are associated with membrane fluidity, acrosome reaction, sperm motility and viability, but limited information exists about the fatty acid profile of ram semen. Our aim was to determine the fatty acid composition in ram spermatozoa and seminal plasma. Sixty ejaculates were obtained from three ram (20 ejaculates/ram) using artificial vagina. Ram spermatozoa ( RS) and seminal plasma ( SP) were separated using centrifugation, and the fatty acids were analysed by gas chromatography. Total lipids obtained in ram spermatozoa were 1.8% and 1.6% in seminal plasma. Saturated fatty acid ( SFA) was proportionally major in SP (66.6%) that RS (49.9%). The highest proportions of SFA corresponded to C4:0 ( RS = 16.3% and SP = 28.8%) and C16:0 ( RS = 16.3% and PS = 20%). The most important unsaturated fatty acid ( UFA) was docosahexaenoic acid ( DHA), 44.9% in RS and 31.5% in SP. The profile of fatty acid and their proportions showed differences between spermatozoa and seminal plasma. [ABSTRACT FROM AUTHOR]

Macrophage metalloelastase-12 ( MMP-12), a protein of the matrix metalloproteinase family, is involved in the breakdown of extracellular matrix in normal physiological processes as well as in disease processes. MMP-12 is almost exclusively produced by macrophages and is associated with inflammatory disorders. Giving the fact that inflammation negatively influences ejaculate parameters, we investigated a possible presence and correlation of MMP-12 in seminal plasma with parameters of the ejaculate, especially in leucocytospermic ejaculates. Forty-two patients who presented for semen analysis were assigned into four groups depending on the result of semen analysis according to the WHO guidelines 2010: normozoospermia ( n = 11), OAT ( n = 10), azoospermia ( n = 10) and leucocytospermia (>1 mio. peroxidase-positive cells per ml) ( n = 11). MMP-12 was detected by ELISA and was measurable in nearly all seminal plasma samples. Generally, MMP-12 concentrations were significantly higher in leucocytospermic samples than in nonleucocytospermic ones ( P = 0.001). The MMP-12 levels between the latter nonleucocytospermic groups did not differ. Moreover, MMP-12 levels correlated with the presence of peroxidase-positive leucocytes. No correlation with CD 14 positive monocytes/macrophages was detected. In this study, we demonstrate that MMP-12 is present in seminal plasma and is correlated with inflammatory conditions in human semen and therefore may serve as predictor of ongoing inflammatory processes. [ABSTRACT FROM AUTHOR]

Oxidative stress has been proposed as one of the potential causes for infertility in men. Retinol and α-tocopherol have an important role in the spermatozoa defences against oxidative stress. A method is described here for the simultaneous determination of retinol and α-tocopherol in human seminal plasma with a suitable sample preparation procedure to prevent retinol and α-tocopherol degradation. After adequate sample preparation, the samples were determined by reversed-phase column chromatography with UV detection. The analytical performance of this method was satisfactory. The intra-assay and inter-assay coefficients of variation were below 10%. The recoveries were as follows: 90.7% (CV 8.1%) for retinol and 98.2% (CV 4.8%) for α-tocopherol. No significant differences in both retinol and α-tocopherol concentration between the smokers and nonsmokers (15 ± 7 n m and 1.86 ± 0.29 μ m versus 15 ± 6 n m and 1.93 ± 0.45 μ m) were found. A selective high-performance liquid chromatography method for the determination of retinol and α-tocopherol in human seminal plasma was developed. [ABSTRACT FROM AUTHOR]

The aim of this study was to elucidate the role of llama seminal plasma in the formation of oviductal sperm reservoirs. Female llamas with follicles in the mature phase were mated with a bulbourethral glands-removed male. Females mated with nonbulbourethral glands-removed males were used as control. Oviducts were obtained by surgery 24 h after mating. The uterotubal junction and isthmus were examined by scanning electron microscopy, and mucopolysaccharides were identified by Alcian blue staining. To know the proteins probably involved in sperm reservoir formation, SDS- PAGE of seminal plasma (8% and 18% resolving gel) was made. Spermatozoa only adhered to the oviductal mucosa surface of uterotubal junction of females mated with nonbulbourethral glands-removed males confirming that seminal plasma and, in particular, bulbourethral secretions are related with the oviductal sperm reservoir formation. Histological sections showed sperm in the lumen, immersed in substance, positive for acid mucopolysaccharides. Alcian blue staining of seminal plasma proteins SDS- PAGE showed a band of high molecular weight containing mucopolysaccharides, only present in nonbulbourethral glands-removed males. Bulbourethral glands would secrete at least eight different proteins that most likely participate in the process of sperm storage in the oviduct. [ABSTRACT FROM AUTHOR]

Semen is a heterogenous and complex fluid with different functions, some of them well known, others still obscure. The aim of this study was to investigate the presence of cathepsins B, S and L in human seminal plasma and their possible associations with other semen variables. Cathepsin B, L and S concentrations were measured in seminal plasma from 99 men utilising commercial ELISA kits. Seminal plasma cathepsin B was approximately 70 times higher, while the cathepsin L values were approximately 500 times higher and the cathepsin S values approximately 40 times higher in seminal plasma than in a group of serum samples. The study shows that seminal plasma contains high levels of cathepsins B, L and S. All three cathepsins were also bound to the surface of prostasomes. [ABSTRACT FROM AUTHOR]

Seminal plasma is a potential source of biomarkers for many disorders of the male reproductive system including male infertility. The identification and characterisation of differentially expressed proteins in seminal plasma of man with normal and impaired spermatogenesis can help in the elucidation of the molecular basis of male infertility. We compared the protein expression profiles of seminal plasma from four different groups of men as follows: normozoospermic, asthenozoospermic, oligozoospermic and azoospermic groups, using two-dimensional differential gel electrophoresis (2- D DIGE). We found eight proteins with statistically significant increased expression in azoospermia compared with at least one of the other studied groups. The differentially expressed spots were fibronectin, prostatic acid phosphatase ( PAP), proteasome subunit alpha type-3, beta-2-microglobulin, galectin-3-binding protein, prolactin-inducible protein and cytosolic nonspecific dipeptidase. Notably, PAP was increased in patients with azoospermia compared with that of all other groups. We have observed no statistically significant differences in protein expression between three of the groups: normozoospermic, oligozoospermic and asthenozoospermic. We suggest that the identified panel of proteins in our study especially PAP have a strong potential to be used as azoospermia markers. However, further investigations will be necessary to validate these markers in samples of larger and independent patient cohorts and to clarify their role in the pathogenesis of male infertility. [ABSTRACT FROM AUTHOR]

The active transport of sperm through the female genital tract is a very important factor in the fertilisation process. There is conflicting evidence as to whether seminal plasma has a positive or a negative effect on reproductive performance regarding the sperm transport in the female genital tract. In this study, we investigated if the perfused swine uterus is an appropriate model to study the effect of human seminal plasma on uterine contractility. It was found that repeated application of human seminal plasma caused a significantly higher rise in pressure and frequency of contractions in perfused swine uteri compared to buffer [a 1.8-fold and 1.5-fold pressure rise (Δ t1SP, P = 0.025; Δ t2SP, P = 0.004)] after the first and second application of seminal plasma respectively, and even a 2-fold pressure rise after the third application with a statistical significance of P = 0.007. The pressure rise caused by the seminal plasma over the entire period of application was calculated using a mathematical programme and was represented by the integral of the pressure curve over time. This perfusion model as well as the use of pressure-time integrals is suitable for patho-physiological studies of the uterus. [ABSTRACT FROM AUTHOR]

This work aimed to assess seminal plasma heme oxygenase (HO) enzyme activity in oligoasthenoteratozoospermia (OAT) males with varicocele. Ninety-three men were divided according to their sperm count and clinical examination into: healthy fertile controls ( n = 34), OAT without varicocele ( n = 37) and OAT associated with varicocele ( n = 22). They were subjected to semen analysis and estimation of seminal plasma HO enzyme activity in the form of bilirubin concentration. Seminal plasma HO enzyme activity decreased significantly in OAT cases compared with controls. Seminal plasma HO in OAT cases associated with varicocele decreased significantly compared with OAT cases without varicocele and healthy controls (mean ± SD; 109.2 ± 29.5, 283.6 ± 88.4, 669.5 ± 236.1 nMol bilirubin/mg ptn/min, P < 0.001). There was positive correlation between seminal plasma HO enzyme activity and sperm concentration, per cent of motile spermatozoa, number of motile spermatozoas ml−1 and significant negative correlation with sperm abnormal forms per cent. It is concluded that varicocele has a negative impact on seminal HO enzyme activity. Therefore, improved seminal picture after correcting varicocele repair might be related, in part, to improved HO action(s). [ABSTRACT FROM AUTHOR]

The aim of this study was to evaluate the correlation between the secretory function of the male accessory glands and sperm parameters in normospermic controls and infertile patients. One hundred and fifty-nine men were investigated: they were composed of two groups: normospermic ( n = 37) and infertile ( n = 122) men with altered sperm characteristics. These infertile men were divided into the following groups: asthenozoospermia ( n = 38), teratozoospermia ( n = 40) and asthenoteratozoospermia ( n = 44). The patients underwent semen analysis and measurements of fructose, neutral alpha-glucosidase and citric acid. The level of fructose was significantly decreased in asthenozoospermic and increased in asthenoteratozoospermic men. It was significantly correlated with semen volume, sperm count, motility and morphology. The seminal alpha-glucosidase levels were significantly correlated with semen volume and pH and citric acid was significantly correlated with pH. Thus, alpha-glucosidase and citric acid levels were associated with semen pH. The significant correlation between semen parameters, accessory glands and epididymal functions highlights the relationship between semen and normal genital tract function. [ABSTRACT FROM AUTHOR]

We examined the effects of normal and leucocyte-positive semen on cytokine expression in endometrial epithelial cell cultures. Cytokines in pooled seminal plasma (SP) samples of men with normal semen parameters without ( n = 9) and with leucocytospermia (LCS) ( n = 9) were determined. Cultures of epithelial endometrial cells of women ( n = 7) in the secretory phase were incubated with 10% SP for 24 h. Cytokines in culture supernatants and mRNA concentrations in the cultured cells were determined. Mean concentrations of interleukin (IL)-1β and transforming growth factor (TGF)-β1 were significantly higher ( P < 0.05) in culture media with SP of men with normal semen parameters and LCS compared with control. Accordingly, a significant increase ( P < 0.05) in mRNA concentrations (amol ml−1) of IL-1β and TGF-β1 could be detected. The mean TGF-β1 and IL-1β concentrations in the culture supernatant with 10% SP from patients with normal semen parameters were 24-fold ( P < 0.05) and 2-fold higher respectively when compared with control. The mean protein concentration of TGF-β1 measured in the supernatant with SP of men with LCS was not significantly reduced as compared with the supernatant with normal SP. In conclusion, our experiments support the concept of an endometrial sensitation effect by semen. [ABSTRACT FROM AUTHOR]

This work aimed to assess heme oxygenase (HO) enzyme activity relationship with different human semen parameters. One hundred and twenty men were divided according to their sperm count and clinical examination into: obstructive azoospermia ( n = 20), nonobstructive azoospermia (NOA) ( n = 25), oligozoospermia ( n = 35) and normozoospermia ( n = 40). Semen analysis, western blot for HO-1 and HO-2, and estimation of seminal plasma HO enzyme activity chemically in the form of bilirubin concentration were carried out. Seminal plasma HO enzyme activity was very low in OA specimens, low in NOA, moderate in oligozoospermia while higher in normozoospermia (mean ± SD; 6.26 ± 2.2, 81.4 ± 35.5, 283.8 ± 90.1, 657.4 ± 227.6 pmol ml−1 min−1) with significant differences. Western blot analysis demonstrated HO-2 expression in all studied groups whereas HO-1 was highly expressed in fertile normozoospermic group compared with other groups. There was positive correlation between seminal plasma HO enzyme activity and sperm concentration, sperm motility percentage, motile spermatozoa ml−1 and sperm normal morphology per cent. It is concluded that HO enzyme activity in the human seminal plasma is related to spermatogenesis and sperm-motility processes. [ABSTRACT FROM AUTHOR]

Leptin is an adipocyte-secreted protein that participates in the regulation of energy homeostasis. Eighty men were investigated; fertile normozoospermia as a control (n = 30) and infertile oligozoospermia (n = 50). The patients underwent estimation of body weight (kg), height (cm), calculation of body mass index (BMI), semen analysis, serum leptin and testosterone hormones. Mean body weight was significantly higher in infertile oligozoospermia compared with controls. Mean height, BMI and serum testosterone levels showed nonsignificant differences between the two groups. Infertile oligozoospermia had significantly higher mean serum leptin level than controls (mean ± SD; 6.88 ± 8.65, 16.3 ± 13.98 ng ml-1, P < 0.01). Serum leptin demonstrated significant positive correlation with age, body weight, BMI and significant inverse correlation with serum testosterone. It had nonsignificant correlation with the height and sperm concentration. These results are suggestive of a link between the adipocyte derived hormone, leptin and male reproduction. [ABSTRACT FROM AUTHOR]

This work aimed to assess seminal α-1,4-glucosidase activity in infertile oligoasthenozoospermic men associated with and without scrotal varicocele. Eighty men were investigated. They were divided into three groups: group 1 ( n = 20), fertile normozoospermic men; group 2 ( n = 30), oligoasthenozoospermia with varicocele; and group 3 ( n = 30), oligoasthenozoospermia without varicocele. The patients underwent medical history, clinical examination, conventional semen analysis and estimation of seminal plasma α-1,4-glucosidase activity by double-beam spectrophotometer method and serum testosterone by radioimmunoassay method. There was a significant decrease in the mean seminal α-1,4-glucosidase activity levels in infertile men versus controls (mean ± SD; 7.66 ± 0.433, 2.088 ± 0.565, 5.384 ± 0.85 mU ml−1 respectively). Mean serum testosterone levels demonstrated nonsignificant differences between studied groups. Seminal α-1,4-glucosidase activity levels demonstrated significant correlation with sperm count, sperm motility percentage and serum testosterone in oligoasthenozoospermia with varicocele group and demonstrated nonsignificant correlation in other groups. It is concluded that varicocele-induced hypoxia is the adverse effect that causes both oligoasthenozoospermia and decreased seminal α-1,4-glucosidase levels. [ABSTRACT FROM AUTHOR]

This work aimed to assess the relationship of seminal ascorbic acid levels with smoking in infertile males. One hundred and seventy men were divided into four groups: nonobstructive azoospermia [NOA: smokers ( n = 20), nonsmokers ( n = 20)]; oligoasthenozoospermia [smokers ( n = 30), nonsmokers ( n = 20)]; asthenozoospermia [smokers ( n = 20), nonsmokers ( n = 20)] and normozoospermic fertile men [smokers ( n = 20), nonsmokers ( n = 20)]. The patients underwent medical history, clinical examination, conventional semen analysis and estimation of ascorbic acid in the seminal plasma calorimetrically. There was a significant decrease in the mean seminal plasma ascorbic acid levels in smokers versus nonsmokers in all groups (mean ± SD; 6.03 ± 2.18 versus 6.62 ± 1.29, 7.81 ± 1.98 versus 9.44 ± 2.15, 8.09 ± 1.98 versus 9.95 ± 2.03, 11.32 ± 2.15 versus 12.98 ± 12.19 mg dl−1 respectively). Fertile subjects, smokers or not, demonstrated significant higher seminal ascorbic acid levels than any infertile group. Seminal plasma ascorbic acid in smokers and nonsmokers was correlated significantly with sperm concentration ( r = 0.59, 0.60, P < 0.001), sperm motility ( r = 0.65, 0.55, P < 0.001) and negatively with sperm abnormal forms per cent ( r = −0.53, −0.50, P < 0.001). Nonsignificant correlations were elicited with semen volume ( r = 0.2, 0.09) or liquefaction time ( r = 0.03, 0.06). It is concluded that seminal plasma ascorbic acid decreased significantly in smokers and infertile men versus nonsmokers and fertile men, and is significantly correlated with the main sperm parameters: count, motility and normal morphology. Also, cigarette smoking is associated with reduced semen main parameters that could worsen the male fertilizing potential, especially in borderline cases. [ABSTRACT FROM AUTHOR]

Numerous investigations have been conducted on the relationship between cigarette smoking and male infertility, however, the exact molecular mechanisms are not well understood in most of the cases. Few studies have indicated the direct effect of seminal plasma (SP) [in different dilutions with phosphate buffer solution (PBS)] from smokers (SM) on the sperm functional parameters from nonsmokers (non-SM). The aim of this study was to provide evidence that cigarette smoking affects male fertility via altering the sperm quality. Our results indicated that exposure of spermatozoa from the non-SM to the SP from the SM yielded a significant reduction in the sperm motility and acrosome reaction and an elevation in the amount of malondialdehyde (MDA), in a certain time course. Exposure of spermatozoa from the SM to the SP from the non-SM or with PBS resulted in the nonsignificant improvement in the altered sperm functional parameters indicating removal of SM's SP and then subsequent reconstitution with physiological media could be of clinical significance in the various assisted reproductive technologies applied for SM. However, the detrimental effect of SM's SP on non-SM's spermatozoa was prominent. In addition, as spermatozoa in SM's SP are susceptible to peroxidative damages, men with such cells who wish to have children should especially benefit from quitting smoking. [ABSTRACT FROM AUTHOR]

Summary. Leucocytospermia has been associated with reduced sperm motility and decreased capacity for sperm–egg interaction. This effect could be mediated through reactive oxygen species (ROS), which, at high concentrations, induce lipid peroxidation and cellular death. The high impact on sperm capacitation reported in other mammalians should be more accurately assessed in the human because premature activation could affect sperm fertilizing capacity. The aim of this study was to evaluate both the effect of ROS on sperm capacitation and the protective role of seminal plasma. Spermatozoa selected by Percoll gradient were incubated with polymorphonuclear (PMN) granulocytes isolated from blood and activated by phorbol-12-myristate 13-acetate (PMA). Different seminal plasma concentrations were added immediately or after 3-h incubation. Afterwards, ROS production was evaluated by luminescence and sperm capacitation by chlortetracycline stain. In PMN granulocytes and sperm suspensions, the basal ROS production was <32 × 10[sup 3] relative luminescence units (RLU). After stimulation with PMA, the rate of ROS production by PMN increased to 1287 × 10[sup 3] RLU. Incubation of sperm with activated PMN resulted in an increase of sperm capacitation (37% versus 19% in the control). Immediate addition of seminal plasma caused a significant reduction in ROS (P < 0.01) and prevented sperm from capacitating. A higher effect in inhibition of sperm capacitation was observed when seminal plasma had been added after 3-h incubation. The results suggest that human sperm capacitation can prematurely be induced by exogenous ROS and this effect can be reversed by seminal plasma. Thus, human sperm capacitation is another functional parameter that may be affected by nonphysiological ROS production. [ABSTRACT FROM AUTHOR]

Summary. Leptin is a 167-amino acid protein that stimulates gonadotrophin-releasing hormone secretion and exerts indirect effects on the gonads via neuropeptide Y, NPY. Recent research has suggested that leptin may also have an effect on testosterone secretion. To investigate the role of leptin in reproduction, leptin in testicular tissue and seminal plasma was examined in relation to leptin in serum, semen sample qualities and vasectomy. Seminal plasma and serum of 64 infertility patients, and 15 individuals after vasectomy, were assayed for leptin using a competitive ‘in house’ radioimmunoassay. The concentration of leptin in seminal plasma was significantly lower in the ‘normal’ semen sample group than in the ‘pathological’ group (Mean ± SEM; 1.45 ± 0.18 vs. 3.19 ± 0.57 ng ml-1 ; P < 0.05), and showed a significantly negative correlation with percentage of motile spermatozoa (r =-0.46; P =0.0005) and with the velocity straight line, VSL, (r =-0.30; P = 0.029). In contrast, leptin concentration in serum did not show any relationship with the spermiogram parameters. In testicular tissue, leptin was preferentially found within the tubuli seminiferi using anti-leptin polyclonal antibody, Ob A-20 Sc 842. The amount of leptin per ejaculate did not significantly change after vasectomy, and was not correlated to fructose, zinc or neutral alpha glucosidase in seminal plasma (P > 0.05). These results suggest that the amount of leptin in the genital tract, including the tubuli seminiferi, may influence the mechanisms involved in the motility development of spermatozoa. [ABSTRACT FROM AUTHOR]

High-performance liquid chromatography was used for the simultaneous determination of α-tocopherol and retinol in serum and semen of 40 subfertile men. The serum levels of the two vitamins were significantly higher in serum than in semen (3- to 10-fold) (P<0.001). The mean α-tocopherol levels were higher in the serum and semen of men with normal sperm parameters (20±5 and 5±4 μmol L-1) than those with oligozoospermia (10±4 and 3±2 μmol L-1), azoospermia (8±3 and 3±1 μmol L-1) and asthenozoospermia (9±6 and 3±2 μmol L-1) (P<0.002). Mean retinol levels in serum were similar in men with normal sperm parameters (2.4±2 μmol L-1) as in those with defective sperm parameters such as oligozoospermia (2.5±2 μmol L-1) and asthenozoospermia (2.1±1.0 μmol L-1) (P=0.15), but significantly lower in men with azoospermia (1.3±0.3 μmol L-1) (P<0.05). The α-tocopherol:retinol ratio was significantly higher in semen than in serum of men with normal sperm parameters (11.5) and those with asthenozoospermia (10.3) compared with ratios in those with oligozoospermia (8.3) and azoospermia (6.3). This may be related to reduced antioxidant activity in sperm dysfunction as a result of lipid peroxidation, from increased generation of reactive oxygen species. [ABSTRACT FROM AUTHOR]

The determination of Chlamydia antibodies in serum is unsuitable for diagnosing infertility in men. The determination of secretory IgA in seminal plasma would seem to be of greater relevance. Since antibodies against C. trachomalis and C. pneumoniae can he detected, a species-specific test for the diagnosis of inkctions of the male genital tract should he used. Only at relatively high titers (≥1:400) do genus-specific tests agree with the species-specific micro-immunofluorescence test. Whether such values can be taken as a true indicator of a local, chronic infection remains to be investigated. [ABSTRACT FROM AUTHOR]

Keeping the self‐renewal and differentiation of spermatogonial stem cell (SSC) in balance is essential for maintaining spermatogenesis. However, whether the cell death of SSC also plays a vital role in the human being remains unknown. To explore the necroptosis of SSC, the activation marker of necroptosis, phosphorylated mixed lineage kinase domain‐like protein (pMLKL) in testes was evaluated by immunofluorescence staining and Western blot. Meanwhile, a total of 81 semen samples were divided based on the sperm concentration (>15, 10–15, 5–10 and 0–5 million/ml) to study the relationships between pMLKL levels and sperm counts. We found that the pMLKL was increased in the ageing human testes (p < 0.05). Moreover, the seminal pMLKL expression was decreased in groups with sperm concentration 0–5 and 5–10 million/ml when compared with normal sperm concentration in young men (p < 0.05). Further analysis revealed that pMLKL showed an age‐related increased expression in men aged 22–60 years with normal sperm concentration. These data demonstrated that the necroptosis of SSC was important for the spermatogenic function and would raise in advance on the point of testicular hypofunction. In conclusion, the pMLKL may serve as a potential biologically seminal indicator for the spermatogenic function in men. [ABSTRACT FROM AUTHOR]

In South American camelids, sperm survival is low after thawing and poor results are obtained when artificial insemination is performed with cryopreserved semen. The aim of this study was to evaluate the effect of different percentages (10% and 50%) of seminal plasma added prior to the process of cryopreservation and also to evaluate the absence of seminal plasma on llama sperm survival after freezing and thawing. A total of 15 ejaculates from five adult llama males (n = 5; r = 3) were evaluated. A significant decrease in sperm motility, viability, membrane function and intact acrosomes was observed in thawed samples (0%, 10% and 50%) when compared to raw semen. Neither morphology nor chromatin condensation was altered in all thawed samples (p > 0.05), but a significant increase (p < 0.05) in the percentage of spermatozoa with fragmented DNA was observed after thawing all samples compared to raw semen. Higher percentages of total and progressive sperm motility were observed when 0% and 10% of seminal plasma were used compared to 50%. However, no statistical differences were established for sperm viability, membrane function, morphology, acrosome status and DNA quality between thawed treatments. To conclude, neither of the percentages of seminal plasma used showed superiority nor cryoprotective effect on llama sperm survival. [ABSTRACT FROM AUTHOR]

The identification of biomarkers associated with seminal traits could aid in the selection of higher quality ejaculates and benefit the swine industry. The objective of this study was to identify boar seminal plasma proteins associated with sperm motility and morphology. Twenty ejaculates from fifteen adult boars from a commercial boar stud were used for this work. After routine semen collection and analysis, ejaculates were classified into two groups: high‐quality semen (HQS) and low‐quality semen (LQS), based on sperm motility and morphology. Semen samples were processed for seminal plasma separation and analysis by 2D SDS‐PAGE. Total and progressive sperm motility differed between groups (p < 0.001), as well sperm morphology (p < 0.05). The intensity of spots identified as Major seminal plasma PSP‐I (PSP‐I) and cathepsin B (CTSB) was higher in LQS as compared to HQS samples (p < 0.05). Also, PSP‐I was positively associated with major and sperm cauda defects. Sperm motility was negatively correlated with both PSP‐I and cathepsin B. We conclude that high concentrations of Major seminal plasma PSP‐I and cathepsin B in boar seminal plasma are associated with reduced total and progressive sperm motility and low sperm morphology and might be used as biomarkers for semen quality. [ABSTRACT FROM AUTHOR]

Unexplained male infertility (UMI) is a condition in which routine semen analysis fails to detect subcellular sperm dysfunctions. In the present research, a comparative proteomics study of seminal plasma (SP) was conducted in men with unexplained infertility whose female partners had undergone in vitro fertilisation (IVF) treatment to find differences in the SP protein profile. Five UMI men with successful and eight with unsuccessful IVF outcome enrolled in this study. Two‐dimensional polyacrylamide gel electrophoresis (2D‐PAGE) technique was used for protein separation. The differentially expressed proteins were identified using mass spectrometry. Results indicated that at least two different protein spots, including clusterin and epididymal secretory protein E1, were over‐expressed (1.5‐ and 2‐fold change, respectively, p < 0.05) while prostate‐specific antigen was downregulated (0.3‐fold change, p < 0.05) in the successful group as compared with the unsuccessful group. Considering the role of all three identified proteins in the sperm quality, the results of the present study introduced these proteins as new candidate biomarkers for success of IVF in UMI couples. [ABSTRACT FROM AUTHOR]

Microscopically abnormal ( n = 26) semen showed significantly higher levels of malondialdehyde (MDA), protein carbonyl (PC) and protein-bound sialic acid (SA) in seminal plasma as compared with normal semen ( n = 24). The percentage of nonmotile spermatozoa showed significant ( P < 0.01) positive correlation with MDA ( r = 0.5) and PC ( r = 0.49). Sperm counts showed a significant negative correlation with MDA ( r = −0.63, P < 0.001) level of seminal plasma. SA correlated ( r = 0.56, P < 0.01) with MDA. The receiver operating characteristic (ROC) curve of MDA and SA showed that MDA of 3.15 μmol l−1 and SA of 3.85 μmol l−1 were optimum cut-off limits to discriminate abnormal semen from normal. In conclusion, high SA might be a protective response against prevailed oxidative stress in abnormal semen. Seminal plasma MDA and SA may act as potential markers of abnormal semen. [ABSTRACT FROM AUTHOR]