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Poor male fertility significantly affects dairy production, primarily due to low conception rates (CR) in bulls, even when cows are inseminated with morphologically normal sperm. Seminal plasma is a key factor in evaluating the fertilizing ability of bull semen. The extracellular vesicles (EVs) in seminal plasma contain fertility-associated proteins like SPAM1, ADAM7, and SP10, which influence sperm function and fertilizing potential. This study aimed to assess EV-associated fertility proteins in bulls with varying fertility levels and to investigate the effects of seminal plasma EVs (SPEVs) from high-fertility (HF) bulls on the spermatozoa of low-fertility (LF) Sahiwal bulls. Seminal plasma was isolated from the fresh semen of Sahiwal bulls, with four bulls classified as high fertility and three as low fertility. Fertility-associated proteins such as SP10, ADAM7, and SPAM 1 were highly expressed in SPEVs of high-fertility (HF) bulls. PKH26 dye-labeled SPEVs demonstrated significant uptake by spermatozoa at pH 6.8 with ≥ 4 h of co-incubation. Exposure of HF SPEVs to low-fertility (LF) spermatozoa reduced acrosome response, capacitation, and reactive oxygen species (ROS) formation. Our findings suggest that protein repertoires in SPEVs influence sperm activities such as motility, acrosome response, and ROS production. Supplementing LF spermatozoa with HF SPEVs could enhance their functional characteristics, highlighting these proteins as potential resources to modulate cattle bull sperm fertilizing ability. [ABSTRACT FROM AUTHOR]

Seminal plasma (SP) is the main vector of C. trachomatis (CT) during heterosexual transmission from male to female. It has immunomodulatory properties and impacts the susceptibility to HIV-1 infection, but its role has not been explored during CT infection. In the female reproductive tract (FRT), CT infection induces cytokine production and neutrophil recruitment. The role of neutrophils during CT infection is partially described, they could be at the origin of the pathology observed during CT infection. During this study, we developed an experimental in vitro model to characterize the impact of CT infection and SP on endocervical epithelial cell immune response in the FRT. We also studied the impact of the epithelial cell response on neutrophil phenotype and functions. We showed that the production by epithelial cells of pro-inflammatory cytokines increased during CT infection. Moreover, the pool of SP as well as individuals SP inhibited CT infection in a dose-dependent manner. The pool of SP inhibited cytokine production in a dose-dependent manner. The pool of SP altered gene expression profiles of infected cells. The culture supernatants of cells infected or not with CT, in presence or not of the pool of SP, had an impact on neutrophil phenotype and functions: they affected markers of neutrophil maturation, activation and adhesion capacity, as well as the survival, ROS production and phagocytosis ability. This study proposes a novel approach to study the impact of the environment on the phenotype and functions of neutrophils in the FRT. It highlights the impact of the factors of the FRT environment, in particular SP and CT infection, on the mucosal inflammation and the need to take into account the SP component while studying sexually transmitted infections during heterosexual transmission from male to female. [ABSTRACT FROM AUTHOR]

Seminal plasma (SP) is rich in extracellular vesicles (EVs), which are still poorly studied, especially in livestock species. To better understand their functional role in both spermatozoa and endometrial epithelial cells, proper characterization of EVs is an essential step. The objective was to phenotypically characterize porcine seminal EVs (sEVs) using cryogenic electron microscopy (cryo-EM), which allows visualization of EVs in their native state. Porcine ejaculates are released in fractions, each containing SP from different source. This allows characterization sEVs released from various male reproductive tissues. Two experiments were performed, the first with SP from the entire ejaculate (n:6) and the second with SP from three ejaculate fractions (n:15): the first 10 mL of the sperm-rich ejaculate fraction (SRF-P1) with SP mainly from the epididymis, the remainder of the SRF (SRF-P2) with SP mainly from the prostate, and the post-SRF with SP mainly from the seminal vesicles. The sEVs were isolated by size exclusion chromatography and 1840 cryo-EM sEV images were acquired using a Jeol-JEM-2200FS/CR-EM. The size, electron density, complexity, and peripheral corona layer were measured in each sEV using the ImageJ software. The first experiment showed that sEVs were structurally and morphologically heterogeneous, although most (83.1%) were small (less than 200 nm), rounded, and poorly electrodense, and some have a peripheral coronal layer. There were also larger sEVs (16.9%) that were irregularly shaped, more electrodense, and few with a peripheral coronal layer. The second experiment showed that small sEVs were more common in SRF-P1 and SRF-P2, indicating that they originated mainly from the epididymis and prostate. Large sEVs were more abundant in post-SRF, indicating that they originated mainly from seminal vesicles. Porcine sEVs are structurally and morphologically heterogeneous. This would be explained by the diversity of reproductive organs of origin. [ABSTRACT FROM AUTHOR]

Background: Despite the clear clinical diagnostic criteria for necrozoospermia in andrology, the fundamental mechanisms underlying it remain elusive. This study aims to profile the lipid composition in seminal plasma systematically and to ascertain the potential of lipid biomarkers in the accurate diagnosis of necrozoospermia. It also evaluates the efficacy of a lipidomics-based random forest algorithm model in identifying necrozoospermia. Methods: Seminal plasma samples were collected from patients diagnosed with necrozoospermia (n = 28) and normozoospermia (n = 28). Liquid chromatography–mass spectrometry (LC–MS) was used to perform lipidomic analysis and identify the underlying biomarkers. A lipid functional enrichment analysis was conducted using the LION lipid ontology database. The top 100 differentially significant lipids were subjected to lipid biomarker examination through random forest machine learning model. Results: Lipidomic analysis identified 46 lipid classes comprising 1267 lipid metabolites in seminal plasma. The top five enriched lipid functions as follows: fatty acid (FA) with ≤ 18 carbons, FA with 16–18 carbons, monounsaturated FA, FA with 18 carbons, and FA with 16 carbons. The top 100 differentially significant lipids were subjected to machine learning analysis and identified 20 feature lipids. The random forest model identified lipids with an area under the curve > 0.8, including LPE(20:4) and TG(4:0_14:1_16:0). Conclusions: LPE(20:4) and TG(4:0_14:1_16:0), were identified as differential lipids for necrozoospermia. Seminal plasma lipidomic analysis could provide valuable biochemical information for the diagnosis of necrozoospermia, and its combination with conventional sperm analysis may improve the accuracy and reliability of the diagnosis. [ABSTRACT FROM AUTHOR]

This study aimed to determine whether male stress is related to seminal stress biomarkers and pregnancy achievement in women exposed to their partner's seminal plasma (SP) in the intracytoplasmic sperm injection (ICSI) cycle. In this pilot prospective study, 20 couples undergoing ICSI, as well as 5 fertile sperm donors and 10 saliva donors, were investigated. Women were exposed to their partner's SP via unprotected sexual intercourse during the ICSI cycle and intravaginal application on the day of ovum pick-up (Day-OPU). Semen samples were collected from male partners by masturbation on the Day-OPU. Saliva and serum samples were collected prior to masturbation. Body fluids were frozen at − 80 °C until assayed. Biomarkers of activity of the sympathetic adrenomedullary axis (salivary alpha-amylase and adrenaline), sympathetic neural axis (noradrenaline and dopamine), hypothalamic–pituitary–adrenal (HPA) system (cortisol), and immune system (C-reactive protein and interleukin (IL)-18) were estimated to examine their association with SP composition and clinical pregnancy achievement. The clinical pregnancy rate was 45.0%. In the unsuccessful ICSI group, blunted levels of salivary and serum cortisol were found compared to the successful ICSI group and the fertile sperm donors. With regard to seminal markers, decreased cortisol level and elevated noradrenaline, noradrenaline/cortisol ratio, and lL-18 levels were strongly associated with ICSI failure (areas under the ROC curves were, 0.813, 0.848, 0.899, and 0.828, respectively). These findings confirm that stress response systems activity affects SP composition, which in turn is associated with ICSI outcomes in women exposed to their partner's SP during an ICSI cycle. [ABSTRACT FROM AUTHOR]

Male infertility is a global public health issue, but studies on the correlation between the dietary components and sperm quality showed inconclusive results due to the heterogeneous population with different dietary habits and environmental stimuli. Herein, the correlation of dietary macro- and micro-mineral intake was evaluated with quality/quantity and oxidant/antioxidant status of seminal fluid in infertile compared to the healthy men. One hundred twenty men attending to the infertility clinic of Ayatollah Mousavi Hospital in Zanjan City were enrolled. Seminal fluid was extracted, and groups were categorized into the infertile (non-standard) and normal (standard) groups based on the WHO, 2020 criteria. Food frequency questionnaire was completed. Seminal malondialdehyde (MDA) and total antioxidant capacity (TAC) were measured by ELISA kit based on the manufacture's instruction. An independent sample t-test was used to determine differences between the two groups, and linear regression model was used to determine the effect of each dietary macro/micro mineral intake on these parameters. Adjusting for all parameters, dietary selenium increased 3.7-folds the seminal TAC level (p=0.04) and decreased sperm with non-progressive motility by 2.4-folds (p=0.04). Higher manganese intake increased the sperm count by 7.8-folds (p=0.005). Dietary copper decreased sperm vitality and increased sperm with slow motility (OR= −1.7, 95% CI= −59.8, −9.9; p=0.007). Dietary zinc (OR=1.24, p=0.01) and iron (OR=1.5, p=0.02) showed a positive effect on sperm vitality. None of macro and micro minerals showed a significant effect on the seminal MDA level. Daily intake of adequate amounts of micro and macro minerals improves sperm quality and increases the antioxidant capacity of the seminal fluid; however, copper showed a negative correlation that must be evaluated in future studies. [ABSTRACT FROM AUTHOR]

A new anodic stripping voltammetry technique has been proposed for direct determination of zinc in seminal plasma of bulls. Two types of electrodes were used as working electrodes—a stationary glassy carbon electrode (GCE) and a thick-film-modified graphite electrode (TMGE) for comparison. The method is rapid, simple, and highly accurate, has high selectivity and low limits of detection (LODs) and limits of quantification (LOQs), and no sample pretreatment steps are also required. The data obtained in both cases agree well with each other and with the data obtained by the independent method of inductively coupled plasma optical emission spectrometry. The relative error of the zinc determination by anodic stripping voltammetry does not exceed 10%. The LOD and LOQ values obtained under optimized conditions were 3.0 and 8.0 µg L−1 for GCE and 2.0 and 5.0 µg L−1 for TMGE, respectively. In contrast to the GCE application, the TMGE application did not include the addition of toxic soluble mercury salts (II) in the electrolyte solution. The zinc content in seminal plasma of bulls is 6.1 ÷ 13.0 mg L−1. The mean values of zinc concentrations in the samples were 8.4, 11.9, and 9.0 mg L−1 for Red, Simmental, and Black-motley animals, respectively. All of these values are within physiological limits and agree well with literature data. [ABSTRACT FROM AUTHOR]

Identifying the metabolome of human seminal plasma (HSP) is a new research area to screen putative biomarkers of infertility. This case–control study was performed on HSP specimens of 15 infertile patients with teratozoospermia (defined as normal sperm morphology < 4%) and 12 confirmed fertile normozoospermic men as the control group to investigate the seminal metabolic signature and whether there are differences in the metabolome between two groups. HSPs were subjected to LC–MS-MS analysis. MetaboAnalyst5.0 software was utilized for statistical analysis. Different univariate and multivariate analyses were used, including T-tests, fold change analysis, random forest (RF), and metabolite set enrichment analysis (MSEA). Teratozoospermic samples contained seventeen significantly different amino acids. Upregulated metabolites include glutamine, asparagine, and glycylproline, whereas downregulated metabolites include cysteine, γ-aminobutyric acid, histidine, hydroxylysine, hydroxyproline, glycine, proline, methionine, ornithine, tryptophan, aspartic acid, argininosuccinic acid, α-aminoadipic acid, and β-aminoisobutyric acid. RF algorithm defined a set of 15 metabolites that constitute the significant features of teratozoospermia. In particular, increased glutamine, asparagine, and decreased cysteine, tryptophan, glycine, and valine were strong predictors of teratozoospemia. The most affected metabolic pathways in teratozoospermic men are the aminoacyl-tRNA, arginine, valine-leucine, and isoleucine biosynthesis. Altered metabolites detected in teratozoospermia were responsible for various roles in sperm functions that classified into four subgroups as follows: related metabolites to antioxidant function, energy production, sperm function, and spermatogenesis. The altered amino acid metabolome identified in this study may be related to the etiology of teratozoospermia, and may provide novel insight into potential biomarkers of male infertility for therapeutic targets. [ABSTRACT FROM AUTHOR]

The aim of this study was to examine the biochemical components and the parameters of antioxidant protection in the seminal plasma (SP) of boars, as well as their relationship with semen quality parameters. Thirty-six boars were included in the study, whose ejaculates were divided into two groups: Group I (good quality semen, > 70% progressively motile sperm, < 20% spermatozoa with abnormal morphology, n = 16), and Group II (poor quality semen, < 70% progressively motile sperm, > 20% spermatozoa with abnormal morphology, n = 20). Significantly higher concentrations of ionized calcium (iCa), total cholesterol (TC), lactate-dehydrogenase (LDH) activity, as well as significantly higher values ​​of antioxidant protection parameters: thiol groups (-SH), paraoxonase 1 (PON1), and total antioxidant capacity (TAC)) were found in the good quality semen, while higher phosphorus (P) concentrations and increased alkaline-phosphatase (ALP) activity were found in the semen of poor quality. A negative correlation of total and progressive sperm motility with P and ALP was found in all examined semen samples, while a positive correlation was found with PON1 and TAC. The percentage of fast sperm cells positively correlated with iCa, chlorine (Cl), lactate, LDH and TAC, while a negative correlation was found with P, magnesium (Mg) and the enzyme creatine-kinase (CK). The percentage of immobile sperm positively correlated with P and ALP, and negatively correlated with TC, CK, PON1 and TAC. Elevated values ​​of PON1 and TAC in SP and a positive correlation with sperm motility indicate the possible use of these parameters as sensitive biomarkers of boar semen quality. To the best of our knowledge there are no published data on association between PON1 activity and boar semen quality. [ABSTRACT FROM AUTHOR]

The metabolomic approach has recently been used in the assessment of semen quality and male fertility. Additionally, the crucial roles of branched-chain amino acids (BCAAs) and aromatic amino acids (AAAs) in metabolic syndrome (MetS) were reported. However, little information exists about the association between BCAAs and AAAs with semen parameters, particularly in men with and without MetS. Our objective was to explore the association between BCAAs and AAAs in blood and seminal plasma and sperm parameters in men with MetS (MetS +) and without MetS (MetS-). In a cross-sectional study between January-July 2022, we investigated 98 men (age: 25–42 years; MetS + : n = 28 and MetS-: n = 70) at Royan Institute, Tehran, Iran. All participants underwent anthropometric indices measurements using standard protocols. From each participant, a single fasting blood sample was collected on the same day that the semen sample was collected. The BCAAs and AAAs in blood and seminal plasma were measured using high-performance liquid chromatography (HPLC). The aromatase activity (total testosterone/ estradiol ratio) was significantly lower in MetS + (0.16) than MetS- (0.35) (p = 0.016). Semen parameters were similar between the MetS + and the MetS- groups. All BCAA and AAA family members, except tryptophan, were higher in the blood plasma of men with metabolic syndrome. Meanwhile, the seminal plasma of BCAA and AAAs were similar. Intriguingly, Valine in blood (r = -0.329; p < 0.001) and seminal (r = -0.237; p < 0.05) plasma were correlated with abnormal sperm morphology in patients without metabolic syndrome (MetS-). Further research is necessary to validate these findings and to explore the underlying mechanisms and interactions between the plasma BCAAs and AAAs and sperm parameters. [ABSTRACT FROM AUTHOR]

Seminal plasma proteins are the major extrinsic factors that can modulate the sperm quality and functions. The present study was carried out to compare the proteomic profiles of seminal plasma from breeding bulls producing good and poor quality semen in an effort to understand the possible proteins associated with semen quality. A total of 910 and 715 proteins were detected in the seminal plasma of poor and good quality semen producing bulls, respectively. A total of 705 proteins were common to both the groups, in which 380 proteins were upregulated and 89 proteins were downregulated in the seminal plasma of poor quality semen, while 236 proteins were co-expressed. The proteins negatively influencing sperm functions such as CCL2, UQCRC2, and SAA1 were among the top ten upregulated proteins in the seminal plasma of poor quality semen. Proteins having a positive role in sperm functions (NGF, EEF1A2, COL1A2, IZUMO4, PRSS1, COL1A1, WFDC2) were among the top ten downregulated proteins in the seminal plasma of poor quality semen. The upregulation of oxidation–reduction process-related proteins, histone proteins (HIST3H2A, H2AFJ, H2AFZ, H2AFX, HIST2H2AB, H2AFV, HIST1H2AC, HIST2H2AC, LOC104975684, LOC524236, LOC614970, LOC529277), and ubiquinol–cytochrome-c reductase proteins (UQCRB, UQCRFS1, UQCRQ, UQCRC1, UQCRC2) indicate deranged oxidation–reduction equilibrium, chromatin condensation and spermatogenesis in poor quality semen producing bulls. The expression of proteins essential for motile cilium (CCDC114, CFAP206, TEKT4), chromatin integrity (PRM2), gamete fusion (IZUMO4, EQTN), hyperactivation, tyrosine phosphorylation, and capacitation [PI3K–Akt signalling pathway-related proteins (COL1A1, COL2A1, COL1A2, SPP1, PDGFA, NGF)] were down regulated in poor quality semen producing bulls. [ABSTRACT FROM AUTHOR]

The purpose of this study was to find out whether the seminal testosterone and/or estradiol levels could serve as prognostic criteria for normal spermatogenesis and whether they are able to characterize the sperm pathology. The study involved healthy young male volunteers (n=269); serum and seminal steroid hormones were measured; the sperm concentration, mobility, and morphology were evaluated. The results indicate that the seminal testosterone concentration is lower (p<0.05) and the seminal estradiol is higher than the corresponding parameters in the serum (p<0.05). The seminal testosterone and estradiol concentrations negatively correlated with the sperm concentration, and the seminal estradiol concentration was higher in pathozoospermic than in normospermic men (p<0.05). It is assumed that the seminal estradiol level can be an indicator of sperm quality and serve as a biological predictor of normal spermatogenesis; in addition, this parameter can be used for diagnostic purposes in patients with impaired spermatogenesis induced by excess of estrogens. [ABSTRACT FROM AUTHOR]

We studied the content of neuron-specific enolase (NSE) in 69 paired samples of blood serum and seminal plasma from men with azoospermia (n=11) and oligoastenozoospermia (n=10) and from men with fertile ejaculate (n=48). NSE concentration was determined by ELISA (Vector-Best kit). The median concentration and the interquartile range of the NSE content in seminal plasma were 65.7 (47.9; 83.4) ng/ml and 24.33 times (р<0.000001) exceeded those for blood serum 2.7 (1.45; 4.0) ng/ml. There were no differences in the content of NSE between the groups for both seminal plasma and blood serum. The content of NSE in seminal plasma did not correlate with the content of NSE in blood serum, and also did not depend on the content of spermatozoa. A weak negative correlation (r=-0.341; p=0.0057) was found between the age of the examinees and the level of NSE in seminal plasma, but not in blood serum. [ABSTRACT FROM AUTHOR]

The cause of endometriosis, which is characterized by the existence of functional endometrial tissue outside the uterine cavity, is poorly understood. Seminal plasma (SP) is rich in multiple cytokines that may promote endometrial tissue survival. Here, we evaluated the effect of SP on growth of endometrial mesenchymal stem cells (MSCs) from women with endometriosis (E-MSCs) and women without endometriosis (NE-MSCs). Proliferation, cell foci formation, cell cycle progression, and growth marker expression of E- and NE-MSCs were promoted by SP. These effects may be mediated through activation of transforming growth factor beta 1 (TGF-β1), Akt, and p42/44 signaling, which enhances CDK2 and CDK6 expression and accelerates cell cycle progression. Xenografts exposed to SP exhibited a three-fold increase in volume and four-fold increase in weight after 14 days. Our findings demonstrate that TGF-β1 in SP may promote endometrial tissue survival which will allow us to understand the pathogenesis and develop novel approaches for prevention and therapies of endometriosis. [ABSTRACT FROM AUTHOR]

Purpose: Previous studies have suggested the involvement of serum insulin-like growth factors (IGFs) and IGF binding proteins (IGFBPs) in the regulation of the female reproductive system. Little is known of these peptides in the seminal plasma (SP) of men and their potential effects on fertility. We assessed SP levels of these peptides in infertile men with low sperm motility (asthenozoospermic; AZ) and low sperm counts (oligozoospermic; OZ), its effects on in vivo sperm motility, and whether there is a correlation with aging. Methods: Twenty-eight infertile men (AZ; n = 18 and OZ; n = 10) and 20 fertile normozoospermic (NZ) men were studied. Seminal plasma IGF-I, IGF-II, IGFBP-2, IGFBP-3, and prostate-specific antigen (PSA) levels were measured, and spermatozoa mRNA transcript patterns were examined. Results: Asthenozoospermic men had higher SP IGF-I, IGF-II, IGFBP-2, and PSA levels than NZ and OZ men, whereas SP IGFBP-3 levels were similar between the three groups. Sperm count positively correlated with SP IGF-I, IGF-II, and IGFBP-2; sperm motility negatively correlated with SP IGF-II and IGFBP-2; and age correlated positively with SP IGF-II. The expression of IGF-I and IGF-II mRNA and mRNA receptors was detectable, but no variations in transcript levels were noted. Conclusion: Decreased sperm motility, but not sperm count, in infertile AZ men is associated with increased SP IGF-I, IGF-II, IGFBP-2, and PSA levels. Changes in SP IGFs and their interactions with IGFBPs and IGF receptors, and PSA levels suggest a role of these SP peptides in modulating sperm motility and possibly prostate disease development in aging men. [ABSTRACT FROM AUTHOR]

The plasma glycoprotein afamin has been previously identified as an alternative carrier protein for vitamin E in extravascular fluids such as plasma and cerebrospinal, ovarian follicular, and seminal fluids. However, to date, no study has established a relationship between afamin levels and infertility in women or men. The purposes of our study were (i) to assess the level of afamin in serum and seminal fluids in infertile men compared to healthy controls and (ii) to study the association between polymorphisms in afamin genes and male infertility. This observational, prospective study evaluated the afamin levels in serum and seminal fluids from infertile men (n = 39) and compared them to those in healthy controls (n = 30). We studied the association between single-nucleotide polymorphisms (SNPs) in the 5'-untranslated region (5'-UTR) of the afamin gene and infertility and analyzed a total of 1000 base pairs from the untranslated region of the afamin gene. Subjects with low sperm motility and low sperm concentration had higher median seminal afamin (18.9 ± 2.9 ng/mg of proteins) and serum afamin concentrations (24.1 ± 4.0 ng/mg of proteins) than subjects with normal sperm parameters (10.6 ± 1.4 ng/mg of proteins) (p < 0.02) (15.6 ± 1.4 ng/mg of proteins) (p < 0.002). A total of five different polymorphisms were found, including one deletion and four single-nucleotide polymorphisms (SNPs). A new transversion (A/T) (position 4:73481093) was identified in an oligoasthenoteratozoospermic patient and was associated with high levels of afamin in plasma and seminal fluids. The prevalence of this variant in our study in the case homozygous for TT is 0.985 (98.5%), and in the case heterozygous for TA is 0.015 (1.5%). Our results suggest that genetic variations in afamin might be associated with male infertility. These findings could significantly enhance our understanding of the molecular genetic causes of infertility. [ABSTRACT FROM AUTHOR]

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. [ABSTRACT FROM AUTHOR]

We have previously shown that high level of seminal interleukin (IL)-18 is positively associated with a greater risk of pregnancy failure in women exposed to their partners' seminal plasma (SP) during the in vitro fertilization (IVF)/intracytoplasmic sperm injection (ICSI) cycle. Since IL-18 and IL-1β considered to be the key immune markers of stress, here we ask whether their increase in SP may be due to the stress experienced by men engaged in the IVF programs. Therefore, we correlated seminal IL-18 with IL-1β and both cytokines with the seminal steroids, whose increase indicates the activation of neuroendocrine stress response systems. Retrospective analysis of stored seminal samples was performed. Based on previously identified cutoff level for content of IL-18 per ejaculate, samples with high IL-18 content from IVF failure group (n = 9), as well as samples with low IL-18 content from IVF success group (n = 7), were included in the study. Seminal cytokines were evaluated using FlowCytomix™ technology. A set of 16 biologically active steroids in SP was quantified by liquid chromatography coupled with mass spectrometry. Concentrations and total amounts per ejaculate of cytokines and steroids were determined. A positive significant correlation was found between the levels of IL-18 and IL-1β. There was also a positive correlation between IL-18 or IL-1β and 17-α-hydroxypregnenolone, 17-α-hydroxyprogesterone, dehydroepiandrosterone (DHEA), DHEA sulfate (DHEAS), androstenedione, testosterone, dihydrotestosterone, progesterone, corticosterone, 11-deoxycorticosterone, and the ratio of DHEAS/cortisol. We suggested that stress-related overexpression of immune and hormonal factors in SP may be the key link between male stress and embryo implantation failure. [ABSTRACT FROM AUTHOR]

Testicular heat stress affects sperm quality and fertility. However, the chronology of these effects is not yet fully understood. This study aimed to establish the early sequential effects of heat stress in bull sperm quality. Semen and blood samples of Nellore breed bulls were collected and distributed into control and testicular heat stress (scrotal bags/96 h) groups. Semen samples were evaluated for sperm motility, abnormalities, plasma membrane integrity, acrosomal membrane integrity, mitochondrial membrane potential, sperm lipid peroxidation, seminal plasma lipid peroxidation, and DNA fragmentation. Blood plasma was also evaluated for lipid peroxidation. An increase in sperm abnormalities was observed 7 days following heat stress. After 14 days, sperm lipid peroxidation increased and mitochondrial membrane function, sperm motility, and plasma membrane integrity decreased. Heat stress effects were still observed after 21 days following heat stress. An increase in sperm DNA fragmentation was observed as a late effect after 28 days. Thus, the initial effects of heat stress (i.e., increasing sperm abnormalities and lipid peroxidation) suggest the presence of oxidative stress in the semen that alters mitochondrial function, sperm motility, plasma membrane integrity, and belatedly, DNA fragmentation. Although sperm abnormalities persisted and increased over time, sperm lipid peroxidation, in turn, increased only until 21 days after heat stress. In this regard, these findings provide a greater understanding of the chronological effects of experimentally induced heat stress on bovine sperm, providing valuable insights about spermatogenesis during the first 28 days following heat stress. [ABSTRACT FROM AUTHOR]

This study aimed to investigate the effect of fractionated seminal plasma on characteristics of common carp Cyprinus carpio cryopreserved sperm. Nanosep® centrifugal devices yielded four seminal plasma fractions with different total protein content ranging in molecular weight from less than 17 to almost 74 kDa. Each protein fraction was added to semen extender medium prior to freezing. Spermatozoon motility characteristics and DNA integrity were analyzed in supplemented and non-supplemented cryopreserved samples. The cryopreservation process strongly affected the swim-up sperm quality. Treatment with fractions 1, 2, 3, and 4 was associated with significantly higher spermatozoon motility rate and curvilinear velocity than seen in extender only, with highest values obtained with fraction 4 (78.21 ± 2.41% and 168.05 ± 4.46 μm/s, respectively). Significantly less DNA damage, expressed as percent tail DNA (12.23 ± 1.27) and olive tail moment (0.68 ± 0.12), was recorded in fraction 4. The findings indicated that addition of fractionated seminal plasma to cryopreservation medium can preserve the quality of common carp sperm. The protective effect of each fraction varied, suggesting the presence of distinct components exerting different effects on cryopreserved sperm function. [ABSTRACT FROM AUTHOR]

MicroRNAs are small, non-coding, single-strand oligonucleotides which regulate gene expression. There is little evidence in the literature about their role in azoospermia and no studies have investigated their presence in the seminal plasma of men with Klinefelter syndrome. This retrospective study investigated if there were any differences in microRNA expression (miR-509-5p, miR-122-5p, miR-34b-3p, miR-34c-5p) in the seminal plasma of patients with obstructive azoospermia, non-obstructive azoospermia and Klinefelter syndrome. Hormone levels were also investigated to identify any correlations with microRNA expression. We analysed 200 subjects (40 Klinefelter syndrome, 60 non-obstructive azoospermia with a normal karyotype, 60 obstructive azoospermia and 40 who were normozoospermic). All subjects underwent semen examination. Total RNA was obtained from seminal plasma and microRNA expression was analysed by RT-qPCR. There was a significant reduction in the expression of all investigated miRNAs in the seminal plasma of all patient categories in comparison with controls. There was a weak negative correlation between FSH values and miR-509-5p expression in non-obstructive azoospermic patients (r = − 0.391; p = 0.014). We hypothesize that in non-obstructive azoospermia and Klinefelter syndrome patients, the downregulation of microRNAs may be caused by damage to the germ cells and aberrant spermatogenesis. In our opinion the identification of seminal plasma microRNAs deriving almost exclusively from the testes could be essential for the development of specific biomarkers for male infertility. The expression of such microRNAs, in combination with hormone values, could comprise testicular markers of abnormal spermatogenesis and failed mature sperm production. [ABSTRACT FROM AUTHOR]

In this study, the concentration/activity of the immune components in the turbot (Scophthalmus maximus) seminal plasma at four time points with the intervals of 2 weeks has been summarized. The results showed that the concentrations of transferrin (TF), transforming growth factor-βs (TGF-βs), complement 3 (C3), IgM and interferon-1β (IL-1β) varied as experiment went on; it was the same with the activities of protease, lactate dehydrogenase and lysozyme. The correlation analysis showed that C3 and IgM were most positively related to motility and straight-line velocity (VSL)/curvilinear velocity (VCL), whereas protease was most negatively correlated with motility and VSL/VCL. This study highlighted the importance in the necessity and possibility in improving the sperm quality during the actual production. [ABSTRACT FROM AUTHOR]

A number of viruses causing sexually transmissible diseases are transmitted via mammalian seminal plasma. Several components of seminal plasma have been shown to influence those viruses and their physiological impact. To unravel whether components of seminal plasma could affect viruses transmitted via other pathways, it was investigated here whether the bovine seminal plasma protein PDC-109, belonging to the Fn-type 2 protein family, influences the activity of influenza A viruses, used as a model for enveloped viruses. We found that PDC-109 inhibits the fusion of influenza virus with human erythrocyte membranes and leads to a decreased viral infection in MDCK cells. In the presence of the head group of the phospholipid phosphatidylcholine, phosphorylcholine, the inhibitory effect of PDC-109 was attenuated. This indicates that the impact of the protein is mainly caused by its binding to viral and to erythrocyte membranes thereby interfering with virus-cell binding. Our study underlines that Fn-type 2 proteins have to be considered as new antiviral components present in mammalian seminal plasma. [ABSTRACT FROM AUTHOR]

Seminal plasma (SP) is thought to be a crucial factor which affects the expansion of regulatory T cells (Tregs) in female reproductive tract during embryo implantation. We propose that seminal transforming growth factor (TGF) β1 is responsible for local accumulation of circulating Tregs, which manifests as changes in Treg frequency in peripheral blood, whereas seminal interleukin (IL) 18 interferes with TGF-β1-dependent cellular reactions. The purpose of the present study is to determine whether the frequency of circulating Tregs is associated with the levels of seminal cytokines and pregnancy establishment in women exposed to partner's SP during in vitro fertilization (IVF)/intracytoplasmic sperm injection (ICSI) cycle. Twenty-nine women were exposed to SP via timed intercourse before the day of ovum pickup (day-OPU) and also subjected to intravaginal SP application just after OPU. Measurements of seminal TGF-β1 and IL-18 were made by FlowCytomix technology. The percentage of CD4+CD25+CD127low+/– Tregs among total circulating CD4+ T cells was determined by flow cytometry and the difference between Treg values on the day of embryo transfer and day-OPU was calculated. The percentage of Tregs on the day-OPU, identified as a predictive factor of clinical pregnancy after IVF/ICSI, showed a positive correlation with IL-18 concentration and content of this cytokine per ejaculate (P <.001 and P <.004, respectively) and negative correlation with the TGF-β1/IL-18 ratio (P <.014).These findings indicate that the adverse effect of seminal IL-18 excess on implantation may be realized by the prevention of postcoital TGF-β1-related migration of circulating Tregs, which clearly manifests as elevated level of Treg frequency in peripheral blood. [ABSTRACT FROM AUTHOR]

In mammals, small RNAs enclosed in exosomes have been identified as appropriate signatures for disease diagnosis. However, there is limited information on exosomes derived from seminal plasma, and few studies have reported analyzed the composition of exosomes and enclosed small RNAs in fish. The half-smooth tongue sole (Cynoglossus semilaevis) is an economically important fish for aquaculture, and it exhibits sexual dimorphism: the female gender show higher growth rates and larger body sizes than males. Standard karyotype analysis and artificial gynogenesis tests have revealed that this species uses heterogametic sex determination (ZW/ZZ), and so-called sex-reversed pseudo-males exist. In this study, we successfully identified exosomes in the seminal plasma of C. semilaevis; to the best of our knowledge, this is the first report of exosomes in fish seminal plasma. Analysis of the nucleotide composition showed that miRNAs were dominant in the exosomes, and the miRNAs were sequenced and compared to identify signature miRNAs as sexual biomarkers. Moreover, target genes of the signature miRNAs were predicted by sequence matching and annotation. Finally, four miRNAs (dre-miR-141-3P, dre-miR-10d-5p, ssa-miR-27b-3p, and ssa-miR-23a-3p) with significant differential expression in the males and pseudo-males were selected from the signature candidate miRNAs as markers for sex identification, and their expression profiles were verified using real-time quantitative PCR. Our findings could provide an effective detection method for sex differentiation in fish. [ABSTRACT FROM AUTHOR]

Element concentrations in serum and seminal plasma were studied in Duroc boars with different semen quality characteristics. Based on the utilization rate of 2174 ejaculates from June to August in 2016, a total of 166 Duroc boars were allocated into three groups: low utilization rate group (LG, 0 to 60% utilization rate), medium utilization rate group (MG, 60 to 80%), and high utilization rate group (HG, 80 to 100%). Serum and seminal plasma samples were collected, and element levels were analyzed using inductively coupled plasma mass spectrometry. The results showed that LG boars had higher concentrations of serum copper and seminal plasma cadmium compared with MG and HG boars (P < 0.05), and serum copper and seminal plasma cadmium were negatively correlated with sperm motility, while positively correlated with the abnormal sperm rate. We observed the abnormal sperm rate increased by approximately 4.53% with serum copper increasing from 1.63 to 2.44 mg/L, while sperm motility decreased by approximately 2.85% with seminal plasma cadmium increasing from 0 to 0.82 μg/L. Moreover, serum iron and manganese levels in the LG group were significantly reduced compared with the HG boars (P < 0.05), and the two elements were negatively correlated with the abnormal sperm rate (P < 0.05). In conclusion, excessive copper and absence of iron and manganese in serum as well as higher seminal plasma cadmium may reduce the utilization rate of semen by impairing sperm motility and morphology, indicating the importance of adding and monitoring microelements in boar diet. [ABSTRACT FROM AUTHOR]

We analyzed cobalt (Co), chromium (Cr), and lead (Pb) concentrations in human semen and catalase CAT activity in seminal plasma and the effects of their relations on the sperm quality. We obtained semen samples from men (n = 168) undergoing routine infertility evaluation. Studies included two groups based on the ejaculate parameters: I (n = 39; normal ejaculate; normozoospermia); II (n = 129; pathological spermiogram). We examined relationships and differences between Co, Cr, and Pb concentrations in seminal plasma, CAT activity, and semen parameters. We did not establish differences in Co, Cr, and Pb concentrations and CAT activity from men between normozoospermic and those with pathological spermiogram. We found a significantly lower Co concentration and CAT activity in males with normal sperm motility than in asthenozoospermic males. We found significantly lower Co and a higher Pb concentration in males with normal morphology of spermatozoa than in teratozoospermic males. We found a significantly higher Pb concentration in the individuals with consumption of alcohol than in those without consumption. There were significant correlations between Co and Pb concentrations, sperm progressive motility (A + B, i.e., fast and slow progressive motility; Co—negatively; Pb—positively), and normal morphology of spermatozoa (Co—negatively; Pb—positively). We found a significant negative correlation between Cr concentration and slow progressive motility, and between CAT activity and volume of ejaculate. Co, Cr, and Pb levels and CAT activity were related to sperm characteristics and male fertility. The impact of alcohol may be manifested by a disturbance in Pb equilibrium in the body. Co and Pb influence progressive motility and normal morphology of human spermatozoa. Thus, Co and Pb levels in semen may be a useful diagnostic in male infertility. Most of the results of this study are in contrast to expectations. Namely, Pb is a toxic element and its harmful effects (poor semen quality) may be expected already at relatively low level of Pb exposure and are particularly visible with increasing of Pb. Co and Cr(III) are essential elements and harmful effects may be expected at their deficiency and/or overexposure. [ABSTRACT FROM AUTHOR]

Purpose: To study if short-term exposure (2 h and 6 h) of endometrial/endometriotic tissues and cells to 10% seminal plasma (SP) can induce EMT/metaplasia.Methods: Basic research experimental study was carried out in a University hospital-based fertility center. Semen samples, peritoneal fluid (PF) from endometriosis patients, endometrial biopsy from premenopausal women, immortalized endometriotic epithelial cell line (12Z), and immortalized endometrial stromal cell line (St-T1b) were studied. Rapid stain identification test (RSID), TGFβ1 immunofluorescence of washed sperms, TGFβ1-ELISA of SP and PF, in vitro study (2 h and 6 h incubation) and real-time PCR of endometrial tissue and cell lines to analyze gene expression of EMT/metaplasia markers and mediators were done.Results: SP is still detectable in washed semen. TGFβ1 was expressed on the plasma membrane of the sperms and was significantly more concentrated in SP (88.17 ng/ml) than PF. 10% SP induced an up-regulation of alpha smooth muscle actin expression in endometrial tissue (p = 0.008) and in 12Z cells (p = 0.05), mostly TGFβ1-independent. TWIST expression was persistently significantly down-regulated while Snail1 and 2 were up-regulated, though insignificant.Conclusion: Our results provide novel evidence to support that even in semen washed twice, SP is still detectable. The changes in EMT/metaplasia markers and mediators give a new insight into a possible effect of SP on the pathogenesis of endometriosis. [ABSTRACT FROM AUTHOR]

Aim: The purpose of the study was to evaluate the impact of seminal plasma in human ejaculates on the sperm DNA quality and DNA longevity.Methods: Semen samples for this study were obtained from 20 donors with a normal spermiogram. Following centrifugation, the sperm pellet was resuspended either in the seminal plasma proceeding from its respective donor, or in an equal amount of PBS, adjusting the concentration to 50 × 106/ml. Each set of samples was incubated at 37 °C for 24 h and the sperm DNA damage was assessed using the chromatin-dispersion test following 0, 2, 6, and 24 h of incubation.Results: Sperm DNA fragmentation (SDF) did not differ between the two experimental conditions at T0; however, Kaplan-Meier estimates to test for survival showed an statistical significant increase of SDF in the seminal plasma group when compared to the PBS group following all timeframes (p 0.000). With respect to sperm DNA longevity, the most dramatic loss of sperm DNA quality occurred during the first 2 h of incubation, with the rate of SDF (rSDF) in the seminal plasma being 2.1 more intense than in PBS. Interestingly, the rSDF was found to vary significantly between individuals, which was confirmed with significant correlations in all rSDF timeframes (rSDF T0-2, p 0.049; rSDF T2-6, p 0.000; rSDF T6-24: p 0.000).Conclusions: Co-incubation of semen samples in seminal plasma after ejaculation increases iatrogenic sperm damage. This effect is statistically significant after 2 h of co-incubation. Subsequently, for ART purposes seminal plasma must be quickly removed after ejaculation-liquefaction, to avoid a higher susceptibility of sperm DNA towards fragmentation. [ABSTRACT FROM AUTHOR]

Purpose: The plasminogen/plasmin system is an important extracellular protease system whose function has been implicated in male reproductive function. However, its clinical relevance to fertility in human assisted reproduction technologies has not been systematically investigated. Here, we examined whether total and active populations of urokinase-type plasminogen activator (uPA) in human seminal plasma and spermatozoa are predictive of pregnancy outcome in couples undergoing insemination or intracytoplasmic sperm injection (ICSI).Methods: Seminal samples from 182 men, 5 donors, 21 patients attending the clinic for infertility screening, and 156 for assisted reproduction technology (ART) treatment (insemination and ICSI), were evaluated. Total uPA in seminal plasma and spermatozoa as well as active uPA in seminal plasma were measured by ELISA. Sperm quality parameters and fertility outcomes following insemination or ICSI were correlated with the uPA values.Results: Active uPA in seminal plasma was positively correlated to the volume of the ejaculate, total number of spermatozoa in the ejaculate, and total motility. However, these values were not prognostic of fertility outcomes. Total uPA in spermatozoa was inversely related to sperm concentration, total sperm in ejaculate, morphology, and total and progressive motility, and this measure was not related to fertility. Importantly, however, higher values of total uPA in seminal plasma were detected in cases that resulted in pregnancy compared to those that did not follow insemination and ICSI treatment.Conclusions: Taken together, these findings lay the foundation for further understanding the mechanism by which total uPA in seminal plasma affects fertility and how this marker can be used as a predictor of ART outcomes. [ABSTRACT FROM AUTHOR]

Purpose: The aim of this study is to evaluate which cryopreservation protocol, freezing before or after swim-up, optimizes cryopreservation outcomes in terms of motile sperm count, motility, morphology, and viability, and also to establish whether sperm viability could be assessed based on sperm motility.Methods: Fifty-three fresh and 53 swim-up prepared samples were considered for the first experiment. In parallel, total motility evaluation by CASA system (computer-assisted sperm analyzer) and hypoosmotic swelling test (HOS-test) was performed in each sample to compare the viability results of both methods. In the second experiment, 21 normozoospermic semen samples and 20 semen samples from male factor patients were included. After fresh ejaculate evaluation, the semen sample of each patient was divided into two aliquots, one of them was frozen before swim-up and the other was frozen after swim-up. Motility, sperm count, morphology, and viability were evaluated after thawing.Results: A linear regression model allows prediction of HOS-test viability results based on total motility: HOS = 1.38 + 0.97 · TM (R2 = 99.10, residual mean squares = 9.51). Freezing before sperm selection leads to higher total and progressive motility, total motile sperm count, and viability rates than when sperm selection is performed before freezing (P < 0.005 in all cases). In fact, sperm selection prior to freezing reaches critical values when subfertile patients are considered.Conclusions: To conclude, total motility evaluation can predict HOS-test viability results, resulting in a more objective and less time-consuming method to assess viability. In addition, sperm freezing prior to swim-up selection must be considered in order to achieve better outcomes after thawing, especially in patients presenting poor sperm baseline. [ABSTRACT FROM AUTHOR]

The caprine arthrite encephalite (CAE) is a disease that affects especially dairy goat. The virus shows compartmentalization features, that allows it to hide at certain times during the course of the disease, making it difficult to control. The present study was conducted to identify the major seminal plasma protein profile of goats infected by CAE and its associations with seroconversion using Western blotting. Two groups containing five males each, were used in this experiment. The first group was composed by seropositive animals and the control by seronegative confirmed by Western blotting and PCR. The semen was collected through artificial vagina and after that, two-dimensional electrophoresis and MALDI-TOF MS were used. Seventy-five spots were identified in the goat seminal plasma gels, equivalent to 13 different proteins with more expression. The similar proteins found in both groups and related to reproduction were spermadhesin Z13-like, bodhesin and bodhesin-2, Lipocalin, protein PDC-109-like, and albumin. In infected goats, proteases such as arisulfatase A have been identified, whose function probably is related to metabolism control of sulfatides, involved to virus control. The other ones were bifunctional ATP-dependent dihydroxyacetone kinase/FAD-AMP lyase, cathepsin F isoform X1, disintegrin and metalloproteinase domain-containing protein 2-like isoform X1, clusterin, carbonic anhydrase 2, electron transfer flavoprotein subunit beta, and epididymal secretory glutathione peroxidase. The results of this study show the reaction of the innate immune system against chronic infection of goats by CAE. [ABSTRACT FROM AUTHOR]

Various microRNAs from the miR-371-3 and miR-302a-d clusters have recently been proposed as markers for testicular germ cell tumours. Upregulation of these miRNAs has been found in both the tissue and serum of testicular cancer patients, but they have never been studied in human seminal plasma. The aim of this study was, therefore, to assess the differences in the expression of miR-371-3 and miR-302a-d between the seminal plasma and serum of testicular cancer patients, and to identify new potential testicular cancer markers in seminal plasma. We investigated the serum and seminal plasma of 28 pre-orchiectomy patients subsequently diagnosed with testicular cancer, the seminal plasma of another 20 patients 30 days post-orchiectomy and a control group consisting of 28 cancer-free subjects attending our centre for an andrological check-up. Serum microRNA expression was analysed using RT-qPCR. TaqMan Array Card 3.0 platform was used for microRNA profiling in the seminal plasma of cancer patients. Results for both miR-371-3 and the miR-302 cluster in the serum of testicular cancer patients were in line with literature reports, while miR-371and miR-372 expression in seminal plasma showed the opposite trend to serum. On array analysis, 37 miRNAs were differentially expressed in the seminal plasma of cancer patients, and the upregulated miR-142 and the downregulated miR-34b were validated using RT-qPCR. Our study investigated the expression of miRNAs in the seminal plasma of patients with testicular cancer for the first time. Unlike in serum, miR-371-3 cannot be considered as markers in seminal plasma, whereas miR-142 levels in seminal plasma may be a potential marker for testicular cancer. [ABSTRACT FROM AUTHOR]

Purpose: To evaluate levels of sperm DNA fragmentation and enzymatic antioxidant status in seminal plasma of Tunisian fertile and infertile men in order to assess the effects of seminal oxidative stress on sperm DNA integrity and semen quality. Methods: Semen samples from 100 infertile patients (40 oligoasthenoteratozoospermics, 31 teratozoospermics and 29 asthenozoospermics) and 50 fertile men (controls) were analyzed for DNA fragmentation by TUNEL assay and biochemical parameters. Seminal antioxidant activities (Superoxide dismutase, Glutathione peroxidase and Catalase) and malondialdehyde concentrations were measured spectrophotometrically. Results: Sperm DNA fragmentation and malondialdehyde levels in infertile groups were more elevated than controls. Nevertheless, the activities of the antioxidant enzymes were significantly lower in abnormal groups compared to normozoospermics. Sperm DNA fragmentation was closely and positively correlated to malondialdehyde levels ( r = 0.37, P = 0.008); meanwhile, reduced seminal antioxidant profile was negatively associated to sperm DNA fragmentation. Interestingly, we noted also that sperm DNA fragmentation was negatively correlated to sperm motility ( r = −0.54, P < 0.001) and positively associated to the abnormal sperm morphology ( r = 0.57, P = 0.002). Conclusions: This report revealed that increased sperm DNA fragmentation can be due to the impaired seminal enzymatic antioxidant profile and increased Lipid peroxidation. Our results sustain that the evaluation of sperm DNA fragmentation and seminal oxidative biomarkers in infertile men is recommended as a consistent prognostic tool for male infertility assessment. [ABSTRACT FROM AUTHOR]

Studies suggest a relationship between semen quality and the concentration of trace elements in serum or seminal plasma. However, trace elements may be linked to DNA and capable of altering the gene expression patterns. Thus, trace element interactions with DNA may contribute to the mechanisms for a trans-generational reproductive effect. We developed an analytical method to determine the amount of trace elements bound to the sperm DNA, and to estimate their affinity for the sperm DNA by the ratio: R = Log [ metal concentration in the sperm DNA/ metal concentration in seminal plasma]. We then analyzed the concentrations of 15 trace elements (Al, Cd, Cr, Cu, Hg, Mn, Mo, Ni, Pb, Ti, V, Zn, As, Sb, and Se) in the seminal plasma and the sperm DNA in 64 normal and 30 abnormal semen specimens with Inductively Coupled Plasma/Mass Spectrometry (ICP-MS). This study showed all trace elements were detected in the seminal plasma and only metals were detected in the sperm DNA. There was no correlation between the metals' concentrations in the seminal plasma and the sperm DNA. Al had the highest affinity for DNA followed by Pb and Cd. This strong affinity is consistent with the known mutagenic effects of these metals. The lowest affinity was observed for Zn and Ti. We observed a significant increase of Al linked to the sperm DNA of patients with oligozoospermia and teratozoospermia. Al's reproductive toxicity might be due to Al linked to DNA, by altering spermatogenesis and expression patterns of genes involved in the function of reproduction. [ABSTRACT FROM AUTHOR]

Purpose: To verify if the presence of varicocele (grades II and III) with and without seminal alterations, using the 5 centile cutoff values in table A1.1 of the World Health Organization (WHO, 2010) manual, alters the seminal plasma levels of proteins DNASE1 (deoxyribonuclease-1) and IGFBP7 (Insulin-like growth factor-binding protein 7), which are related to apoptosis regulation and cell proliferation, respectively, demonstrating that these proteins are important for correct spermatogenesis. Methods: This cross sectional study was performed at the Sao Paulo Federal University Paulo between May 2014 and April 2016. A total of 61 male adolescents were included in this study, of which 20 controls without varicocele (C), 22 with varicocele and normal semen analysis (VNS) and 19 with varicocele and altered semen analysis (VAS). Seminal plasma from each patient was used for Western blotting analysis of individual protein levels. Values of each protein were normalized to a testicular housekeeping protein (PARK7-protein deglycase DJ-1). Results: Levels of IGFBP7 protein are increased in varicocele. Levels of DNASE1 are progressively decreased in varicocele (lower in varicocele and normal semen analysis, lowest in varicocele and altered semen analysis) when compared to adolescents without varicocele. DNASE1 levels are positively correlated with sperm concentration and morphology (correlation values of 0.400 and 0.404, respectively; p values of 0.001 and 0.001, respectively). Conclusion: In conclusion, in adolescents, seminal plasma levels of IGFBP7, responsible for proliferative activity, are increased in varicocele grades II and III, and DNASE1, responsible for apoptosis regulation, are lower in varicocele, lowest in varicocele and low semen quality. These proteins demonstrate molecular alterations brought upon by varicocele. Moreover, DNASE1 is capable of discriminating a varicocele that causes alterations to semen quality from one that does not. We propose that the initial response of varicocele is to increase proliferative activity which, if followed by regulation of apoptosis, may lead to the ejaculation of a population of sperm that are in accordance with WHO cutoff values but, in the presence of dysregulated apoptosis, leads to lower sperm concentration and morphology. [ABSTRACT FROM AUTHOR]

The present study evaluated the effects of chilled storage and cryopreservation on sperm motion characteristics, antioxidant enzyme activities, and lipid peroxidation in the Pacific cod Gadus macrocephalus. Sperm motility and the activities of superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GPx), glutathione reductase (Gr), and lipid peroxidation (measured via malondialdehyde (MDA) content) were determined after the milt was stored at 4°C for 12 h, cryopreserved without cryoprotectant in 12% propylene glycol (PG), cryopreserved in 12% PG+0.1 mol/L trehalose, or cryopreserved in 12% PG spermatozoa but centrifuged to decant the supernatant prior to cryopreservation (only sperm cells were cryopreserved). After chilled storage or cryopreservation, the SOD, CAT and GPx activities were reduced in sperm cells and increased in seminal plasma in almost all treatments; sperm motility parameters were also decreased. However, the addition of trehalose into the cryoprotectant could significantly improve the postthaw sperm quality as revealed by the sperm average path velocity. This improvement might be attributed to the function of trehalose in scavenging reactive oxygen species. Chilled storage and cryopreservation had significant effects on sperm motion characteristics, antioxidant enzyme activities, and lipid peroxidation in the Pacific cod. [ABSTRACT FROM AUTHOR]

Purpose: This study was undertaken in order to establish a new reference value for the total antioxidant capacity (TAC) in seminal plasma as a predictor of fertility. This study also aims to propose a detailed protocol for the TAC assay including calculation of assay results and assessment of sensitivity and specificity over possible cutoff values in infertile men and controls with proven and unproven fertility. Methods: Seminal plasma from 279 infertile patients and 46 normal healthy men referred to a male infertility testing laboratory were tested to measure TAC by a colorimetric assay kit. Receiver operating characteristics (ROC) curves were generated to establish cutoff values, sensitivity, and specificity, and the distribution of cutoff values in controls and infertile patients was calculated. Results: Infertile patients showed significantly lower levels (mean ± SEM) of total antioxidants (micromolar Trolox equivalents) in their seminal plasma (1863.84 ± 27.16 μM) compared to those from fertile men (2013 ± 56.04 μM, P = 0.019). A preferred cutoff TAC value of 1947 μM could facilitate better diagnosis of oxidative stress (OS) in men with male factor infertility. At this threshold, the specificity of TAC assay was 63.0 % and the sensitivity 59.5 % with a positive predictive value of 90.7 % and a negative predictive value of 20.4 %. Conclusions: Our results establish a new diagnostic cutoff TAC value of 1947 μM in seminal plasma to distinguish prevalence of OS in infertile patients compared to healthy men. This study provides a robust reference value of seminal plasma TAC that may provide an important diagnostic tool to the physicians for managing OS and male factor infertility in such patients. [ABSTRACT FROM AUTHOR]

In this research, the concentration and activity of oxidants and anti-oxidants in turbot semen, and their effects on sperm quality were studied. The results showed that superoxide dismutase (SOD), catalase, glutathione reductase (GR), uric acid, vitamin E (V) and vitamin C (V) were more abundant in seminal plasma than in spermatozoa. The variation for each of them was specific. In seminal plasma, the activity of SOD and GR increased from November 15, November 30 to December 15, and then decreased on December 30. The concentrations of both VC and uric acid decreased during the first 3 sampling times and increased on December 30. The oxidants in seminal plasma accumulated to the highest on December 30. Lactic acid (LA) and ATP levels decreased to the lowest on December 30. The correlation analysis showed that GR had the significant positive relevance to sperm motility and VSL/VCL, while ·OH had negative relevance to them. [ABSTRACT FROM AUTHOR]

Background: Male infertility has largely idiopathic, multifactorial origin. Oxidative stress is a major factor that affects spermatogenesis, in particular in aging. Cerium dioxide nanoparticles (CNPs) due to their antioxidative properties are promising to impact on the development of male infertility. The aims of this study were to investigate the effects of CNPs on fertility parameters in 24-month male rats and to overview relevant literature in the field of personalized treatments, predictive diagnosis, and preventive measures for male health and fertility. Methods: We included 30 24-month-old male rats. After a week of adaptation to the standard diet, the rats were randomly divided into three groups with ten rats in each. Group 1 (controls) received only a standard diet. The rats of group 2 and 3 in adjunct to the standard diet during 10 days received intragastrically 10 % sodium citrate and citrate-coated CNPs in dose 1 mg/kg, respectively. We assessed sex hormones, epididymal sperm parameters and spermatogenesis, ultrasound, and morphological data of rat reproductive organs. Results: After a 10-day administration of CNPs, we revealed significant decrease of lipid peroxidation product levels in serum and increase of catalase and SOD activity, associated with increase of sperm count ( p < 0.001) and improvement in quantitative sperm parameters (motility, viability, and percentage of spermatozoa). We found no significant changes between sperm quantitative parameters in citrate-treated rats and controls and observed age-related decrease of activated Leydig cell number and focal atrophy of the seminiferous tubules. In CNP group, we observed regeneration of seminiferous tubules, increase number and activation of Leydig cells, and 2.5-fold significant increase of serum testosterone. Ultrasound data showed the slight increase of linear measurement and volume of rat testes in CNP group. Review highlights the benefits for predictive diagnosis, preventive measures, and personalized approaches to manage male infertility in the general concept of male health also related to aging. Conclusion: Citrate-coated 2-5-nm CNPs lead to increase in sex hormones levels, sperm count, and quality, as well as the activation of spermatogenesis in 24-month-old male rats. Nanoceria demonstrated the perspectives to be an effective infertility treatment via reduction of oxidative stress in male reproductive organs, in particular in aging. [ABSTRACT FROM AUTHOR]

Purpose: The aim of this study was to analyze the seminal plasma of patients with idiopathic/male factor infertility and healthy controls with proven fertility by NMR spectroscopy, with a hope of establishing difference in biomarker profiles, if any, between the groups. Methods: A total of 103 subjects visiting the infertility clinic of Manipal University with normozoospermic parameters, oligozoospermia, asthenozoospermia, azoospermia and teratozoospermia were included. Semen characteristics were analysed by standard criteria. Seminal plasma was subjected to NMR spectroscopy at a 700 MHz H frequency. The resultant data was analyzed by appropriate software. Results: The analysis revealed significant differences between the fertile control group and other forms of male infertility. Interestingly, seminal plasma profile of the idiopathic infertility group showed distinct segregation from the control population as well as other infertile groups. The difference in biomarker profiles between the idiopathic infertility and the rest of the groups combined could originate from either the up-regulation or down regulation of a several compounds, including lysine, arginine, tyrosine, citrate, proline and fructose. Conclusion: Our data suggests the presence of a metabolic reason behind the origin of idiopathic infertility. H NMR based metabonomic profiling based on concentration of biomarker lysine has the potential to aid in the detection and diagnosis of idiopathic infertility in an efficient manner. [ABSTRACT FROM AUTHOR]

Purpose: To study whether intravaginal application of seminal plasma after follicle aspiration has the potential to increase implantation and clinical pregnancy rates after IVF-ET. Methods: We conducted a prospective, double-blind, placebo-controlled randomized study of 230 patients undergoing IVF-ET cycles. 500 μL of Fresh seminal plasma from the patient's partner or culture medium (placebo) were injected in the vaginal vault just after follicle aspiration. The main outcome measured was ongoing clinical-pregnancy rate. Results: After ET cancellation in ten patients due to lack of fertilization or embryo cleavage, 220 embryo transfers (103 and 117 in the study and control groups) resulted in a clinical pregnancy rate of 36.9 % and 29.1 % for the study and control groups, corresponding to a relative increase of 26.8 %. After an early pregnancy loss of 13.1 % (5/38) and 23.5 % (8/34) in the study and control groups respectively an ongoing pregnancy rate of 32.0 % (33/103) and 22.2 % (26/117) was achieved corresponding to a relative increase of 44.1 %. Multivariate logistic regression analysis adjusted for study group, age, infertility, and cycle characteristics did not demonstrate any parameter that could predict occurrence of clinical pregnancy rates after IVF-ET. Conclusions: Patients who underwent SP intravaginal insemination after oocyte pick-up reached higher implantation and clinical pregnancy rates following ET compared to controls, although the difference did not reach statistical significance. More studies and variable methodologies may clarify the potential clinical effect of SP in improving live birth rates after ART. [ABSTRACT FROM AUTHOR]

Diamine oxidase (DAO) was purified to homogeneity from human seminal plasma by consecutive chromatographic fractionation on heparin-sepharose, phenyl-sepharose, CIM-QA, and Superdex 200. Human seminal plasma DAO behaves electrophoretically similar to DAO proteins from other human tissues and has very similar enzymatic properties with histamine and aliphatic diamines being the preferred substrates as well as significant conversion of polyamines. The cellular source and functional importance of DAO in human semen remain to be determined. [ABSTRACT FROM AUTHOR]

Purpose: In IVF procedures, endometrial function is a limiting factor of the pregnancy rate and the aims of this study is to determine whether seminal plasma insemination at ovum pick-up in IVF has any effect on pregnancy rate. Methods: We designed a single center, 2 parallel groups, randomized pilot study. All couples undergoing an IVF procedure in our clinic between January 2010 and December 2011 were considered for enrollment in the study. The patients who met the inclusion criteria were randomized into two groups by simple randomization: the seminal plasma group (SP group) and the reference group (noSP group). We applied seminal plasma into the cervix and vaginal vault at the time of the OPU in the patients of the SP group. The primary outcome was the clinical pregnancy rate in the SP group compared with the noSP group and the secondary outcome measure was the implantation rate. Results: 400 patients met the inclusion criteria and were randomized. 54 patients were excluded from the study mainly because they didn't undergo the embryo transfer. Finally, 164 patients were included in the SP group and 182 patients in the noSP group and analyzed. We found a statistically non-significant increase in the pregnancy rate in the SP group (55.5 % versus 44 %, p = 0.09) and a statistically significant increase in the implantation rate (34.7 % versus 27.5 %, p = 0.026). Conclusions: Our results support the idea that SP insemination may have the potential to increase pregnancy rate in IVF procedures but further studies must be carried out. [ABSTRACT FROM AUTHOR]

Purpose: There has been an increasing interest in the evaluation of metal ion concentration, present in different body fluids. It is known that metal ions, especially zinc play vital role in the fertility of human semen. Objective: The main objective of the study is to evaluate the Zn concentration in Normospermia samples by Atomic absorption spectroscopy (AAS) and to predict the same by artificial neural network (ANN). Materials and methods: Normospermia semen samples were collected from the patients who came to attend semen analysis at Bangalore assisted conception centre, Bangalore, India. Semen analysis was done according to World Health Organization (WHO) guidance. Atomic absorption spectroscopy was used to estimate the total Zn in these samples, while the Back propagation neural network algorithm (BPNN) was used to predict the Zn levels in these samples. Results: Zinc concentration obtained by AAS and BPNN indicated that there was a good correlation between the estimated and predicted values and was also found to be statistically significant. Conclusion: The BPNN algorithm developed in this study could be used for the prediction of Zn concentration in human Normospermia samples. Future perspective: The algorithm could be further developed to predict the concentration of all the trace elements present in human seminal plasma of different infertile categories. [ABSTRACT FROM AUTHOR]

In the present review, sperm morphology, acrosome reaction, motility, short-term storage and cryopreservation are summarized and discussed in sturgeon (Chondrostei, Acipenseriformes). The elongated head of spermatozoon comprises an acrosome with 8-12 posterolateral projections. Usually three endonuclear canals are observed in the nucleus. Proximal and distal centrioles and 3-6 mitochondria are located in the midpiece region. The flagellum consists of an axoneme with a typical '9 + 2' structure of microtubules and presents a ribon-like structure due to two lateral membranous fins. Egg water, Ca and Mg can trigger acrosome reaction. Trypsin- and chymotrypsin-like activities are reported in sturgeon sperm. These physiological properties of sturgeon sperm are identified as serine activity with 33 kDa molecular mass and can be inhibited by their respective inhibitors. The K prevents sperm activation in seminal plasma, and hypo-osmolality or decrease of extracellular K triggers sperm activation. Extracellular Ca is involved in flagellar beating pattern and sperm velocity. After activation, sperm motility, velocity, and flagellar beating frequency, wavelength and amplitude decrease, while number of waves and curvature increase. Sturgeon sperm can be stored for several days at 4 °C; however it is better to add K into the immobilizing medium because it prevents sperm activation during incubation. Regarding sperm cryopreservation, methanol is a better cryoprotectant than DMSO. Either short-term storage or cryopreservation of sperm generates damage to spermatozoa that lead to reduction of sperm motility performance. Some studies suggest using an activation medium containing Ca for enhancing sperm motility performance of incubated or frozen-thawed sperm. [ABSTRACT FROM AUTHOR]

Biochemical parameters of seminal and blood plasma in fish are species specific and related together in other aspects. Characterization of these biochemical parameters is essential when evaluating physiological conditions and in determining the effective factors on seminal plasma composition influencing seminal physiology. This study aimed to specify and contrast some seminal and blood plasma biochemical parameters in the wild common carp ( Cyprinus carpio). Total protein, cholesterol, glucose, and ionic parameters (Mg, Ca) of seminal and blood plasma were measured in 15 specimens of the wild common carp ( C. carpio). Results indicated that blood plasma parameters of glucose, cholesterol, and Ca were significantly higher ( P < 0.01) than those of seminal plasma, while no significant differences ( P > 0.05) were observed between the seminal and blood plasma levels for total protein and Mg ion ( P > 0.05). Results of the correlation between biochemical parameters of seminal and blood plasma revealed a significant relationship between Mg ion and glucose levels ( P < 0.01). There were no significant correlations ( P > 0.05) between seminal and blood plasma levels for cholesterol, total protein, and Ca ion. [ABSTRACT FROM AUTHOR]

This study was conducted to investigate the effects of vitamin E, vitamin C, probiotics, dietary protein level, and their combination on semen traits and seminal plasma macro- and microminerals in 65-week-old male broiler breeders after zinc-induced molting. One hundred eighty birds were induced to molt by mixing zinc oxide (3,000 mg/kg) in the diet. The birds were divided into six groups (five replicates) by completely randomized design. One group was kept as control (16% CP), while the other five were supplemented with vitamin E (100 IU/kg feed), vitamin C (500 IU/kg feed) probiotics (50 mg/L), protein level (14% CP), and their combination. Semen samples were weekly collected for determination of semen volume, sperm concentration, motility, and dead sperm percentage. Analyses of Na, K, Ca and Mg, Zn, Fe, Mn, and Cu in seminal plasma were also performed. Overall, mean semen volume was significantly high in vitamin E and C supplemented groups compared to control. Overall mean sperm motility was significantly higher in vitamin E supplemented group, whereas dead sperm percentage was significantly lower in the vitamin C group compared to control. Mineral analyses revealed that overall mean seminal plasma Mg increased significantly in vitamin E and C supplemented groups compared to control. Similarly, significantly high overall mean seminal plasma Cu concentration was observed in vitamins E and C and combination groups. It can be concluded that vitamins have a vital role in improving semen quality and bioavailability of Mg and Cu in seminal plasma of the post-molt cockerels. [ABSTRACT FROM AUTHOR]