Seminal Plasma Levels Research Articles

Anandamide Level in Men with Oligoasthenoteratozoospermia

Background: Male infertility is a relatively common medical condition. Male partners are found to be solely responsible for 20–30% of infertility cases. Oligoasthenoteratozoospermia (OAT) is the most common laboratory finding in infertile men. Endocannabinoids are endogenously produced substances, that primarily act at cannabinoid receptors (CBRs), thus reproducing some of the biological actions of the natural cannabis sativa components (the “cannabinoids”). The endocannabinoid system is involved in several physiological processes, including fertility and body weight control. Objective: The aim of this study was to detect the level of anandamide (AEA) in patients with OAT and its relation to BMI. Patients and methods: This study was a case control study. It was hold between June 2018 to April 2019 in the Dermatology, Venereology and Andrology Department at Aswan University Hospital. The study included 20 patients with OAT and 9 as controls. Semen analysis using CASA was done. AEA level in seminal plasma was measured using ELISA kits. BMI was calculated. Results: AEA level in seminal plasma significantly decreased in men with ligoasthenoterat- ozoospermia than control (P < 0.009). AEA concentration on seminal fluid was significantly increased with an increase in BMI (P= 0.023). On the other hand, a significant negative relationship was present between BMI and semen volume (P= 0.03). Conclusion: AEA concentration in seminal plasma was significantly decreased in patients with OAT. AEA concentration in seminal fluid was significantly increased with an increase in BMI. On the other hand, a significant negative relationship was present between BMI and semen volume.

Association of Copper-to Zinc Ratio with Sperm Concentration among Males Investigated for Infertility

Background: The importance of copper and zinc in the fertility potential of males are well understood. The close relationship and the antagonistic interactions between copper and zinc requires further investigation especially because of conflicting reports in literature on the association of the trace elements with sperm quantity and quality. This study evaluates seminal plasma copper and zinc concentrations, copper-to-zinc ratio and their associations with sperm concentration in infertile males. Methods: This is a cross-sectional study of 400 males investigated for infertility and 100 male control subjects. Seminal plasma copper and zinc were determined using atomic absorption spectrophotometry technique (Buck Scientific Model VGP-210, Germany). The subjects were grouped based on sperm count; normozoospermia(&gt;15 x 106cells/mL), oligozoospermia (&lt;15x106cells/mL) and azoospermia (no sperm cell). The levels of copper, zinc and Cu/Zn ratio were compared using unpaired Students-t-test and analysis of variance while Pearson correlation coefficient was used to assess the correlation between measured variables and sperm count. Results: Seminal plasma zinc was significantly lower (p&lt;0.001) while copper and Cu/Zn ratio were significantly higher (p&lt;0.001) in infertile than fertile males. The Cu/Zn ratio was significantly higher (p&lt;0.001) among infertile males than controls. Seminal plasma levels of copper and Cu/Zn ratio increased with decreasing concentrations of sperm cells while zinc levels increase with increasing concentration of sperm cells.

Leptin is upregulated in epididymitis and promotes apoptosis and IL-1β production in epididymal epithelial cells by activating the NLRP3 inflammasome

Epididymitis, one of the most common urological disease, is a significant cause of male infertility. Leptin is capable of modulating both reproduction and immune response. We analyzed the serum and seminal plasma levels of leptin in infertile patients with or without chronic epididymitis. Experimental epididymitis models were generated by administrating 200 μg Lipopolysaccharide (LPS) to Sprague-Dawley rats. The expression of leptin in epididymis were detected using qPCR, Western blots 6–72 h after injection, and using immunohistochemistry 72 h after injection. Besides, rat epididymal epithelial cells were isolated as an in vitro model and were treated with leptin (5–40 ng/ml, 6–48 h), LPS (1ug/ml, 6 h), and NLRP3 inflammasome inhibitor MCC950 (10 μM, 2 h). Cell Counting Kits-8 assay and Annexin V/PE assay were used to evaluate cell viability and apoptosis. Quantitive PCR and ELISA assay were used to detected inflammatory cytokines interleukin-1beta (IL-1β) production. Western Blots were used to detect molecular related to cell apoptosis, IL-1β maturation, and NLRP3 inflammasome. We found that patients with chronic epididymitis presented a significantly higher level of seminal plasma leptin and correlated declined sperm progressive motility. Leptin and leptin receptor expression in epididymis was significantly upregulated 24 h after LPS administration both in mRNA and protein level, and highly expressed in the epididymis epithelium 72 h after LPS administration. In epididymal epithelial cells, leptin reduced cell viability and promoted apoptosis in a concentration-dependent manner via cleavage of caspase-9, caspase-3, and PARP. Leptin enhanced the LPS-induced production of IL-1β, which was associated with increased IL-1β maturation and caspase-1 activation. Furthermore, NLRP3 inhibitor MCC950 attenuated the effects of leptin or co-treatment with LPS on NLRP3, ASC expression, IL-1β maturation, and caspase-1 activation, which indicated that leptin promotes IL-1β production via activating the NLRP3 inflammasome. These data suggested that leptin may act as a potential evaluation and treatment target for epididymitis and male subfertility.

The role of rabbit seminal plasma natural antioxidants in the realization of spermatozoid activity after ionizing irradiation

Aim. To investigate the effects of X-rays on the content of rabbit seminal plasma L-carnitine and α-tocopherol, as well as physiological parameters of spermatozoid in 10, 30 and 90 days after whole body irradiation of animals in the dose range of 0.1–2.0 Gy. Methods. White rabbits-males Soviet Shinshila were irradiated by X-rays. Physiological parameters of sperm were evaluated by light microscopy, and the content of natural antioxidants was determined using liquid chromatograph “Agilent 1200”. Results. It was shown that low doses of ionizing radiation (0.1 Gy and 0.5 Gy) in 10 days after irradiation lead to the increase of the L-carnitine level in seminal plasma, whereas at 1.0 Gy and 2.0 Gy its concentration decreases. In additional, there is a decrease in the seminal plasma α-tocopherol content with a minimum of 2.0 Gy, although a dose of 0.1 Gy dose not cause changes in the index. The L-carnitine level is no longer different from the control for a dose 0.1 Gy in one month after rabbit irradiation, while the α-tocopherol concentration continues to decrease. Conclusions. The increasing of L-carnitine level in the rabbit’s seminal plasma at 0.1 Gy and 0.5 Gy contributes to increased motility and a radio-stimulating effect on sperm. Radiation-induced depletion of the α-tocopherol pool in semen disappears at a later period.&#x0D; Keywords: ionizing radiation, rabbits, seminal plasma, natural antioxidants

Effect of bioactive peptide on ram semen cryopreservation

This present study investigated the effect of bioactive peptide (BAPT) (BAPT) on the quality of ram semen during cryopreservation. Ram ejaculates were extended with Tris buffer supplemented with no antioxidants (as control group), 20 μg/mL BAPT (as BAPT20 group), 40 μg/mL BAPT (as BAPT40 group) and 60 μg/mL BAPT (as BAPT60 group). After cryopreservation, sperm quality including motility, vitality, the percentage of hypoosmotic swelling test (HOST)-positive spermatozoa and the percentage of intact acrosomes was assessed. Furthermore, the malondialdehyde (MDA) in seminal plasma and spermatozoa were analyzed, followed by the measurement of superoxide dismutase (SOD), catalase (CAT) and glutathione-peroxidase (GSH-Px) levels in seminal plasma. After in vitro fertilization, the embryonic cleavage rates and development rates of different groups were analyzed to compare the developmental abilities of spermatozoa. The results showed that the post-thaw sperm motility was significantly higher in the BAPT60 group compared to those in the BAPT20, BAPT40 and control groups (P < 0.05). The percentage of live sperms significantly increased from 48.12 ± 2.35% for the BAPT20 group, 55.43 ± 2.16% for the BAPT40 group to 57.53 ± 3.15% for the BAPT60 group. The percentage of HOST-positive spermatozoa was significantly higher in the BAPT60 group than those in BAPT20, BAPT40 and control groups (P < 0.05). The MDA levels in seminal plasma and spermatozoa were significantly reduced with BAPT supplement (P < 0.05). Additionally, the SOD, CAT and GSH-Px levels in the BAPT experimental groups were significantly higher than those of the control group, which further indicated that BAPT significantly inhibit the reactive oxygen species (ROS) production during the cryopreservation of ram semen. Furthermore, the embryonic cleavage rates and development rates of the BAPT40 and BAPT60 groups were significantly increased in comparison with the BAPT20 and control groups (P < 0.05).In conclusion, BAPT improved the ram sperm quality via inhibiting the ROS production during cryopreservation, and could be applied as a promising supplement for ram semen cryopreservation.

Associations of toxic and essential trace elements in serum, follicular fluid, and seminal plasma with In vitro fertilization outcomes

Toxic and essential trace elements are associated with human fertility. However, limited studies have been conducted on the relationship between trace elements and in vitro fertilization (IVF) outcomes in the Chinese population. In this exploratory study, 103 couples who underwent IVF/intracytoplasmic sperm injection (ICSI) treatment in our reproductive center were recruited. Serum and follicular fluid (FF) samples and seminal plasma samples were collected from the female and male partners, respectively, on the day of oocyte retrieval. Concentrations of chromium (Cr), nickel (Ni), arsenic (As), selenium (Se), cadmium (Cd), and lead (Pb) in the specimens were measured by inductively coupled plasma-mass spectrometry. Generalized linear models were used to evaluate the associations of toxic and essential trace element levels with the intermediate parameters and outcomes of IVF at both the sex-specific and couple levels. The Cr level in the serum of the female partners was inversely associated with the count of mature oocytes retrieved (p for trend = 0.033). In terms of embryo development, As concentrations in female serum and FF were inversely associated with the probabilities to obtain good-quality cleavage embryos (p for trend < 0.01). There were significant correlations between follicular Se concentrations and seminal As concentrations and higher quality cleavage embryos (p for trend < 0.01). Moreover, a positive correlation was observed between the female serum Se concentrations and blastocyst formation (p for trend = 0.031). In contrast, a higher follicular Se level was negatively correlated with embryo development at the blastocyst stage (p for trend < 0.01). Regarding the clinical outcomes, a positive association of Se levels in seminal plasma and a negative association of follicular Cd levels was observed with the probabilities of pregnancy (p for trend = 0.006 and 0.035, respectively) and live birth (p for trend = 0.014 and 0.027, respectively). The results indicate that exposure to toxic elements (Cr, As and Cd) is associated with intermediate parameters and outcomes of IVF. Furthermore, male Se exposure may be related to better pregnancy outcomes.

Biochemical Assessments of Seminal Plasma Zinc, Testis-Expressed Sequence 101 and Free Amino Acids and Their Correlations with Reproductive Hormones in Male Infertility

The role of the male factors in the couple's infertility has been significantly increased in recent years due to a sententious assessment of male reproductive functions and enhanced diagnostic tools. We investigated the correlations among the seminal plasma (SP) levels of each of zinc, testis-expressed sequence 101 (TEX101), and free amino acids levels with reproductive hormones in adult fertile and infertile men. The study included 100 infertile men categorized into 50 non-obstructive azoospermic patients and 50 patients with idiopathic oligoasthenoteratozoospermia (iOAT), in addition to 50 fertile controls. Semen analyses, serum ELISA assays for male reproductive hormones (follicle-stimulating hormone (FSH), luteinizing hormone (LH), total testosterone, and prolactin), colorimetric assays of SP zinc and total proteins, SP free amino acids using high-performance liquid chromatography (HPLC), and ELISA assays of SP TEX101 were performed for all subjects. Infertile men with azoospermia had significantly lower SP median levels of zinc, TEX101, and many SP free amino acids compared to both men with iOAT and fertile controls (P˂0.05 for all). There were lower SP levels of zinc and some free amino acids among men with iOAT compared to the fertile controls (P˂0.05 for all) with non-significant difference regarding to SP TEX101 (P˃0.05). Azoospermic men exhibited negative correlations between FSH, LH, and prolactin with some SP free amino acids (P˂0.05 for all), and a positive correlation between glycine with total testosterone (P˂0.05). Among iOAT patients, LH and FSH were positively correlated with SP zinc, TEX101, and some measured free amino acids (P˂0.05 for all). Total testosterone was positively correlated with some amino acids, while prolactin was negatively correlated with glycine (P˂0.05 for all). iOAT and azoospermic men exhibited low SP zinc and some free amino acids levels that were more pronounced in azoospermic men and were significantly associated with the reproductive hormones. TEX101 could be a helpful confirmatory test for azoospermia.

Biochemical changes of cryopreserved seminal plasma and spermatozoa of the giant grouper Epinephelus lanceolatus after preservation and transportation using dry-ice

The present study aims to investigate the effects of exposure of the seminal plasma and spermatozoa of the giant grouper Epinephelus lanceolatus to dry ice (−79 °C) during transport on their quality. In all, 15 amino acid compounds were determined. The quantification of total proteins were measured using the Bradford method, and amino acid concentration were measured using the HPLC method. The cryopreserved seminal plasma was transferred from a liquid nitrogen tank to a styrofoam box filled with dry ice. Total protein and amino acids were measured after 24, 48, and 72 h. For comparative purposes, total protein and fifteen compound of amino acid were also measured. Both parameters were also measured after the cryopreserved seminal plasma were immersed in liquid nitrogen after 24 and 48 h exposed to dry ice. The results showed that the exposure of seminal plasma to dry ice for 24, 48 and 72 h during transportation or immersion back into the liquid nitrogen after 24 and 48 h does not change the total protein levels either in seminal plasma or spermatozoa. However, the level of each amino acid compound in the seminal plasma had significantly decreased.

Malondialdehyde Level in Seminal Plasma of Cryopreserved Holstein Bull Semen after Addition of Zinc, Cysteine, Prostaglandin F2α and their Combination in vitro

The study was conducted to know the level of Malondialdehyde (MDA) in seminal plasma of cryopreserved semen of Holstein bulls after addition of zinc sulphate, cysteine, PGF2α and their combination in vitro. Semen was collected from 7 Holstein bulls, presented in Artificial insemination Center which belonged to the Directorate of Animal Resources/ Ministry of Agriculture at Abu-Graib at the west of Baghdad. Pooled semen were diluted with Tris- based extender and divided into five parts. The first part (T1) serve as a control (without addition). The 2nd part (T2) added to it zinc sulphate (0,576 mmol/ ml). The 3rd part (T3) added to it cysteine (5 mmol/ ml). The 4th part (T4) added to it PGF2α (37.5 pg/ ml). while the 5th part added to it a combination of previous substances at the same concentration. They packed in straws and cryopreserved in a liquid nitrogen and after 30, 60 and 90 days. Seminal plasma when isolated to measure the level of MDA. The results showed a significant decrease (P&gt;0.01) in MDA level in the combination treated group (zinc, cysteine and PGF2α) 0.450 ± 0.11 (mmol/ ml) as compared with control group 1.025 ± 0.38 (mmol/ ml), zinc 0.867 ± 0.12 (mmol/ ml), cysteine 1.06 ± 0.12 (mmol/ ml) and PGF2α group 0.968 ± 0.17 (mmol/ ml) respectively. It was concluded from this study that addition of a combination of zinc, cysteine and PGF2α to the Holstein bull semen could decrease the level of MDA which might be due to the synergistic effect of these substances.

Oxidative stress markers in seminal plasma of idiopathic infertile men may be associated with glutathione S-transferase M1 and T1 null genotypes.

This study aimed to investigate the association between glutathione S-transferase (GST) M1 and T1 null genotypes and thiobarbituric acid reactive substances (TBARS), total antioxidant capacity (TAC) and nitric oxide (NO) levels in male infertility. For this purpose, semen samples were collected from fertile and infertile subjects, and then they were genotyped for GSTT1 and GSTM1 genes using multiplex-PCR. The TBARS, TAC and NO levels in seminal plasma were then measured via the ferric-reducing ability of plasma (FRAP). A significant association was observed between GSTT1 null genotype and oligozoospermia, asthenozoospermia and teratozoospermia. But, the GSTM1 null genotype was merely associated with teratozoospermia. Moreover, the GSTT1-/GSTM1+ combined genotype was associated with all subgroups of male infertility. Besides, an association was observed between GSTT1-/GSTM1- genotype and asthenozoospermia and teratozoospermia. Further analysis showed that the GSTT1 null genotype was associated with increased NO in asthenozoospermia. Also, the GSTT1 null genotype was associated with increased TBARS in oligozoospermia and asthenozoospermia. As well, GSTM1 null genotype was associated with decreased TAC and increased NO in asthenozoospermia respectively. As a preliminary conclusion, the GSTM1 and GSTT1 null genotypes could be considered as genetic risk factors for male infertility, interfering with some oxidative stress markers in infertile men.

Men with idiopathic oligoasthenoteratozoospermia exhibit lower serum and seminal plasma melatonin levels: Comparative effect of night-light exposure with fertile males.

Melatonin is a darkness hormone secreted by the pineal gland, which serves a role in idiopathic oligoasthenoteratozoospermia (iOAT). The present study aimed to evaluate the seminal plasma and serum melatonin levels of 50 patients with iOAT and 50 normal fertile controls and the effects of exposure to light at night on semen parameters. Semen analyses were performed according to the World Health Organization 2010 guidelines. Measurements of serum and seminal plasma melatonin, serum TSH, FT3, FT4, free testosterone, prolactin, FSH and LH were performed using ELISA. The overall results revealed that the serum and seminal plasma levels of melatonin were lower in patients with iOAT compared with the control subjects (P=0.0004 and 0.01, respectively). Patients with iOAT who were exposed to light at night exhibited lower serum and seminal plasma melatonin levels compared with those who were not exposed to light at night (P<0.0001 and 0.02, respectively). Additionally, similar significant differences were identified in control subjects exposed to light at night when compared to non-exposed controls. There was a significantly positive correlation between serum melatonin levels and sperm motility in the entire iOAT patient cohort (r=0.614; P<0.0001) and a significantly positive correlation between the serum and seminal plasma melatonin levels in the non-exposed iOAT patient subgroup (r=0.753; P<0.001). Thus, darkness and sleep at night may improve the semen parameters of patients with iOAT, as evidenced by the effects of light exposure at night on the serum and seminal plasma levels of melatonin and, consequently, on semen parameters.

Heterogenic Origin of Micro RNAs in Atlantic Salmon (Salmo salar) Seminal Plasma.

The origin and contribution of seminal plasma RNAs into the whole semen RNA repertoire are poorly known, frequently being overlooked or neglected. In this study, we used high-throughput sequencing and RT-qPCR to profile microRNA (miRNA) constituents in the whole semen, as well as in fractionated spermatozoa and seminal plasma of Atlantic salmon (Salmo salar). We found 85 differentially accumulated miRNAs between spermatozoa and the seminal plasma. We identified a number of seminal plasma-enriched and spermatozoa-enriched miRNAs. We localized the expression of some miRNAs in juvenile and mature testes. Two abundant miRNAs, miR-92a-3p and miR-202-5p, localized to both spermatogonia and somatic supporting cells in immature testis, and they were also highly abundant in somatic cells in mature testis. miR-15c-5p, miR-30d-5p, miR-93a-5p, and miR-730-5p were detected only in mature testis. miRs 92a-3p, 202-5p, 15c-5p, and 30d-5p were also detected in a juvenile ovary. The RT-qPCR experiment demonstrated lack of correlation in miRNA transcript levels in seminal plasma versus blood plasma. Our results indicate that salmon semen is rich in miRNAs, which are present in both spermatozoa and seminal plasma. Testicular-supporting somatic cells are likely the source of seminal plasma enrichment, whereas blood plasma is unlikely to contribute to the seminal plasma miRNA repertoire.

Coenzyme Q10 effect on semen parameters: Profound or meagre?

Coenzyme Q10 has shown promise in treating male infertility; however, there are inconsistencies across the published data. We undertook a quantitative meta-analysis by pooling data from three placebo-controlled randomised clinical trials (RCTs) in order to evaluate the efficacy of CoQ10 in improving semen parameters. Sperm count, sperm motility, sperm forward motility, sperm morphology and CoQ10 level in the seminal plasma were measured and quantitatively correlated with CoQ10 oral administration. Pooled analysis showed a significant impact of CoQ10 in improving sperm motility and forward motility, without a significant impact on sperm count, sperm morphology, ejaculate volume or seminal plasma level of CoQ10. Efficacy assessment suggested that CoQ10 shows better results at higher doses and when administered for a period of more than 3 months but not longer than 6 months. We conclude that CoQ10 has a profound effect on sperm motility and a meagre effect on all other parameters. Therefore, CoQ10 can be used for treating asthenozoospermic infertility with the dosage and duration depending upon the severity of the disorder and the patient's response to the treatment.

CRIPTO and miR-371a-3p Are Serum Biomarkers of Testicular Germ Cell Tumors and Are Detected in Seminal Plasma from Azoospermic Males.

miR-371a-3p is currently the most informative reported biomarker for germ cell tumors (GCTs). Another developmental-related biomarker, CRIPTO, is involved in the regulation of pluripotency and germ cell fate commitment. We aimed to assess the value of CRIPTO as a diagnostic and prognostic biomarker of testicular GCTs (TGCTs) and also to assess its presence in seminal plasma samples, compared with miR-371a-3p. In total, 217 and 94 serum/seminal plasma samples were analyzed. CRIPTO was quantified using ELISA and miR-371a-3p using bead-based isolation followed by RT-qPCR. Methylation profiling (EPIC array) for the CRIPTO promoter region was undertaken in 35 TGCT tissues plus four (T)GCT cell lines. Significantly higher CRIPTO concentration was found in sera of non-seminomas compared to controls (p = 0.0297), and in stage II/III disease compared to stage I (p = 0.0052, p = 0.0097). CRIPTO concentration was significantly positively correlated with miR-371a-3p levels in serum (r = 0.16) and seminal plasma (r = 0.40). CRIPTO/miR-371a-3p levels were significantly higher in seminal plasma controls when compared to serum controls (p = 0.0001, p < 0.0001). CRIPTO/miR-371a-3p were detected both in normospermic and azoospermic males, and levels were higher in TGCTs compared to GCNIS-only. We have provided the largest dataset of evaluation of CRIPTO in serum and seminal plasma of GCTs, showing its potential value as a biomarker of the disease.

Comparative Analysis of PFOS and PFOA Toxicity on Sertoli Cells.

Perfluoroalkyl chemicals induce male reproductive toxicity. Current evidence showed the effects of the chemical exposure on the deterioration of testicular functions, and reduction in epididymal sperm counts. Previous studies showed that PFOA and PFOS displayed a high correlation with each other in seminal plasma levels, but induced different effects on semen variables. In this study, we focused on the comparative toxicity analysis of PFOA and PFOS, using a rat primary Sertoli cell model. Our transcriptomic data showed that PFOA and PFOS treatments (40 μM) perturbed global gene expression. While PFOS induced higher toxicity in affecting cytoskeleton signaling, Sertoli cell-cell junction, and inflammation, underlined by Ingenuity pathway analysis. Immunocytochemical staining revealed that PFOS treatment (40 and 80 μM) induced truncated actin filament and disorganized bundled configuration in the cell cytoplasm. Moreover, disorganized distribution of N-cadherin (N-cad) and β-catenin (β-cat), and defragmentation of ZO-1 at the Sertoli cell-cell interface was evident. At 80 μM of PFOS, cytoplasmic distribution of N-cad, β-cat, and ZO-1 were observed. We then examined whether resveratrol, a polyphenol antioxidant, was able to protect the cells from PFOS toxicity. The pretreatment of Sertoli cells with 10 μM resveratrol prevented the formation of truncated actin filament and dis-localization of β-cat. Western blot analysis showed that Res pretreatment increased the levels of basal ES proteins (N-cad and β-cat), tight junction proteins (ZO-1 and occludin), and gap junction protein, versus control.

Seminal Plasma Clusterin as a Biomarker for Spermatogenesis in Varicocele Patients before and after Varicocelectomy

Abstract Introduction a varicocele is an abnormally dilated pampiniform plexus of veins. Hyperthermia, oxidative stress, and elevated apoptotic index have been described in men with variocele and infertility. Clusterin (Clu) is a chaperone-like glycoprotein which is synthesized by Sertoli cells in high concentrations and is deposited onto the membranes of and mature spermatozoa, giving rise to the possibility that it has a role in sperm development. Secreted form of clusterin is a powerful anti-apoptotic. Aim To explore clusterin level in seminal plasma of infertile men with varicocele in comparison to normal healthy fertile subjects and to compare the seminal plasma clusterin level before and after varicocelectomy Methods The study included 25 patients suffering from varicocele with infertility (whether primary or secondary) for 12 months, and 10 healthy volunteers of matched age and sex as control. For all the patients Sub-inguinal varicocelectomy was done. Semen analysis, and seminal plasma clusterin assay were carried out before and three months after varicocelectomy. Results there was a statistically significant difference between patients and controls as regards sperm count, progressive motility, and seminal plasma clusterin (P &amp;lt; 0.01). Three months after varicocelectomy, the sperm count had a highly significant improvement in the patient’s group (Mean= 17.90 + 8.86) compared to a preoperative count (Mean= 11.43 + 7.56). There was a highly significant improvement in post-operative progressive motility in the patient’s group (Mean= 34.5 + 20.2) compared to a preoperative progressive motility (Mean= 22.05 + 15.5). Post-operative clusterin showed a highly significant elevation in the patient’s group postoperative (Mean= 10.73 + 4.5) compared to a preoperative clusterin (Mean= 6.68 + 3.8). Conclusion Based on our study we could suggest another way of contribution of varicocele to male infertility by affecting seminal plasma clusterin level.

Determination of 27 amino acids’ levels in seminal plasma of asthenospermia and oligospermia patients and diagnostic value analysis

•Amino acids (AAs) are abundant in seminal fluid and play an important role in the reproduction process. However, the functional significance of AAs in seminal is not clear. This study was designed to profile the AAs in seminal fluid of asthenozoospermia (AS) and oligospermia (OL) patients and evaluate its potential diagnostic value. To determine the true function of AAs in seminal fluid, an ultra high performance liquid chromatography-mass spectrometry (UPLC-MS/MS) was developed and validated. The seminal fluid samples collected from 76 AS patients, 58 OL patients, and 76 healthy subjects were determined by UPLC-MS/MS and analyzed by fisher discriminant and receiver operating characteristic (ROC). The results showed that, the AAs levels were dramatically decreased in AS and OL patients compared with healthy subjects. What’s more there were significant variation in the AAs profile of AS and OL patients. Specifically, some polar AAs decreased, while nonpolar AAs increased in OL patients. Based on the level of AAs, 93.4 % of original and 90.9 % of cross-validated grouped cases were correctly classified between AS patients, OL patients, and healthy subjects. The further ROC analysis shows that glutamic acid, tyrosine, aspartic acid, proline, and tryptophan have the diagnostic value (P < 0.01) in the three groups. Moreover, aspartic acid and glutamic acid exhibit a high correlation (Correlation Coefficient >0.9) in AS, OL, and healthy subjects. In conclusion, there are significant variation in AAs profile in seminal plasma of AS and OL patients with high accuracy in fisher classification. The associations between the levels of AAs in seminal fluid of AS, OL, and healthy subjects may have clinical applications in providing valuable diagnostic indicator.

Human spermatozoa of male patients with subfertility express the interleukin-6 receptor.

Male subfertility has been associated with bacterial infections and chronic inflammation. In this context, several studies investigated cytokine levels in seminal plasma, whereas interleukin-6 (IL-6) appears to be crucial. However, little is known about its receptor, the IL-6R expression on human spermatozoa. Thus, the aim of the present study was to screen spermatozoa for IL-6R expression and to identify its localisation. Semen samples of 137 patients (median age 37.69, SD±7.82) with subfertility were analysed. Sperm analysis including determination of IL-6 was performed following the World Health Organization criteria. Also, flow cytometry was performed for sperm IL-6R expression. IL-6R+ cells were used for immunofluorescence staining to identify receptor localisation. The results showed positive staining for IL-6R in the midpiece of spermatozoa. Furthermore, a significant correlation between sperm IL-6R expression, seminal plasma IL-6 and total sperm count could be demonstrated, whereas a negative correlation was observed in sperm IL-6R expression and motility. However, no statistical significance could be observed between IL-6R expression, vitality and morphology. Moreover, incubation of spermatozoa with IL-6 led to a slight but significant decrease in motility after 24hr. These data suggest that IL-6R expression may play a role in impaired sperm function during inflammation.

Effect of Dietary Copper and Zinc Supplementation on Semen Quality of Murrah Bulls

The effect of dietary supplementation of Cu and Zn on semen quality parameters and certain bio-chemicals parameters were evaluated in Murrah bulls. Twelve mature Murrah bulls (4-6 years of age) were divided into three groups (n=4) T1, T2, and T3 based on semen volume and concentration and were fed as per ICAR standard (2013). However, the animals were supplemented with 0%, 25%, and 50% Cu and Zn above the basal diet in T1, T2 and T3 groups, respectively for 180 days. Quantitative and qualitative characteristics of semen, blood and seminal plasma antioxidant status, blood and semen minerals (Cu, Zn, Ca and Mn) were determined in experimental Murrah bulls. Semen ejaculate volume (mL) increased in T2 and T3 while sperm concentration (million/mL), intact acrosome (%), HOST reacted spermatozoa (%) increased in T3 group compared to T1 and T2 groups. No difference was observed in mass motility, pH, live spermatozoa (%) in semen sexual behaviour except dismounting time which was decreased in T3 than T1 and T2 (plessthan0.05). SOD activity in blood plasma and LPO activity decreased in seminal plasma in both T2 and T3 groups than T1, whereas catalase activity did not show any significant difference. Cu and Zn supplementation in T2 group improved plessthan0.05) Zn level in blood and seminal plasma and Cu level in blood plasma only, but not in seminal plasma. Therefore, it can be concluded that supplementation of Zn and Cu at 50% above the recommended levels of ICAR (2013) improved the qualitative and quantitative attributes of semen in Murrah bulls.

Relationships between metabolic status, seminal adipokines, and reproductive functions in men from infertile couples.

Adipokines could be a link between metabolic syndrome (MS) and infertility. While the association between circulating adipokines and fertility has been extensively studied in females, this relationship in males was less investigated, although some adipokines are detectable in seminal plasma (SP). The aim of this study was to determine adipokine levels in blood and SP and to assess the relationships between adipokines, MS and semen parameters in men from infertile couples. Male partners of infertile couples referred to four medical French centers were enrolled in years 2013-2016. Subjects (n = 160) aged 18-45 years were assessed for anthropometric, biochemical, sperm, and circulating hormonal parameters. Leptin, adiponectin, resistin, chemerin, visfatin, and IL-6 were measured in serum and SP. Infertility duration was higher in men with than without MS. Adipokine concentrations were higher in blood than in SP, except for IL-6 and visfatin. The most striking result was the significant correlation observed between seminal IL-6 and spermatozoid concentration, progressive motility, and sperm vitality. Moreover, while men with MS exhibited an expected lower adiponectinemia, they displayed 2.1-fold higher adiponectin levels in SP than men without MS. Finally, logistic regression analysis showed that BMI, infertility duration, and adiponectin serum/SP ratio were independently associated with MS. These results suggest an involvement of seminal adipokines to modulate fertility in men with MS and that seminal IL-6 could play a beneficial role on sperm functionality. Further mechanistic studies are necessary to investigate the precise roles of these adipokines in male reproduction.