Seminal Plasma Levels Research Articles

Cobalt's role in modulating antioxidant systems and semen quality in males.

The present study investigated associations between environmental exposure to cobalt (Co) and the levels of oxidative stress parameters and the antioxidant defense system in the seminal plasma of fertile males. The study population consisted of 117 healthy, non-smoking, fertile men from the southern region of Poland. The study was carried out in 2021–22. Based on the median cobalt levels in seminal plasma, subjects were divided into two groups: those with low (Co-L) and high (Co-H) cobalt concentrations. Semen parameters assessed according to WHO 2021 recommendations. After the analysis of spermiograms, observed reduction in progressive motility after 1hour was found in the Co-H group. Moreover, superoxide dismutase (SOD), glutathione S-transferase (GST) in the Co-H group had lower activity and GR higher activity. The OSI (Oxidative stress index) were higher in the group with high cobalt concentration in semen. The concentrations of redox balance parameters: TOS, TAC and OSI significantly were higher in the Co-high group as well as GR activity. Environmental exposure to cobalt decreases sperm motility in both normal and abnormal semen. The findings from this study affirm that cobalt can induce oxidative stress and alter oxidative stress markers in semen.

The association between levels of samarium, hafnium, tungsten and rhenium in seminal plasma and the risk of idiopathic oligo-astheno-teratozoospermia in men of childbearing age.

Oligo-astheno-teratozoospermia (OAT) is a global public health problem, which affects 30% men of childbearing age. Meanwhile, with the rapid development of industry and economy, the contents of rare earth elements (REEs) in the environment are increasing. However, little is known about the associations between REEs levels and OAT risk. To evaluate the associations between the levels of four REEs (samarium (Sm), hafnium (Hf), tungsten (W), rhenium (Re)) in seminal plasma and OAT risk, from October 2021 to November 2022, semen samples from 924 men of childbearing age (460 controls and 464 cases) were collected from the reproductive center of the First Affiliated Hospital of Anhui Medical University. Inductively coupled plasma-mass spectrometry (ICP-MS) was used to measure the levels of Sm, Hf, Re and W in seminal plasma. Bayesian kernel machine regression (BKMR) was conducted to explore the joint effects of levels of four REEs in seminal plasma on the risk of OAT and select the one exerting a major role; generalized linear regression models (GLM) with log link function were employed to investigate the association of every REE level in seminal plasma and OAT risk; sankey diagram and linear regression models were utilized to describe the associations between the levels of four REEs and the indexes of sperm quality. The levels of four REEs in seminal plasma were higher in the case group than levels in the control group (pSm = 0.011, pHf = 0.040, pW = 0.062, pRe = 0.001, respectively). In BKMR analysis, theOAT risk increased when the overall levels of four REEs were higher than their 55th percentile compared to all of them at their 50th percentile, and Re level played a major role in the association. Additionally, Re level in seminal plasma was positively associated with theOAT risk in the single element model after adjustment of covariates (medium vs. low: OR (95% CI) = 1.55 (1.10, 2.18); high vs. low: OR (95% CI) = 1.69 (1.18, 2.42)). Lastly, the sankey diagram and linear regression models revealed that Sm level was negatively associated with the PR%, total sperm count and total progressively motile sperm count; Hf level was negatively associated with the PR%; W and Re levels were negatively associated with the PR% and total motility, and Re level was positively associated with abnormal morphology rate. Men of childbearing age with OAT had higher levels of Sm, Hf and Re in seminal plasma than those in the control group. An increasing trend for theOAT risk was observed with an increase in mixture levels of Sm, Hf, W and Re, and Re exposure level played a major role in the association whether in BKMR model or single element model. Additionally, the levels of these four REEs were negatively associated with the indexes of sperm quality.

Phosphatidylserine Levels in Seminal Plasma of Iraqi Infertile Men Using Direct and Indirect Swim Up Sperm Preparation Techniques

Phosphatidylserine (PS), exposed on viable sperm, is required for fertilization as it is recognized by specific receptors located on the microvilli of the oocyte to promote sperm: egg fusion. The heads of viable and motile sperm exhibit PS. With (PS) exposure progressively increasing during sperm transit through the epididymis. PS asymmetry on the leaflets of plasma membrane is arranged in a complex way, via multiple proteins functioning in opposite directions; further, PS is also essential for many intracellular trafficking events. The aim of this study was to evaluate the phosphatidylserine PS level in seminal plasma of infertile men before and after activation using Direct Swim Up (DSU) and Indirect Swim Up (IDSU) activation method and make a comparison between them. Forty infertile men were included in this study during their attendance to the infertility clinic at High Institute for Infertility Diagnosis and Assisted Reproductive Technologies, Al-Nahrain University. Normozoospermic or mild asthenozoospermic semen samples were used to choose the cases, according to WHO manual standards (1999 -2010). The collected semen samples obtained were divided into three parts: The first part prepared was as an in vitro sperm characterization before activation while the second and third parts were after activation using direct and indirect swim up techniques respectively. The level of seminal plasma phosphatidylserine was increased after the activation techniques from the baseline level before activation. Comparison of Seminal plasma phosphatidylserine levels between direct and indirect sperm activation technique were significantly higher in direct swim up activation technique than indirect one. Phosphatidylserine (PS) in semen plasma of infertile men was significantly increased after activation using Direct Swim Up DSU preparation method.

The Association Between the Levels of Oxidative Stress Indicators (MDA, SOD, and GSH) in Seminal Plasma and the Risk of Idiopathic Oligo-asthenotera-tozoospermia: Does Cu or Se Level Alter the Association?

Epidemiological studies on the associations between the levels of oxidative stress (OS) indicators (MDA, SOD, and GSH) in seminal plasma and the risk of idiopathic oligo-asthenotera-tozoospermia (OAT) are still inconsistent. Additionally, whether the associations can be altered by the status of essential trace elements is still unknown. To investigate the relationship between MDA, SOD, and GSH levels in seminal plasma and the risk of idiopathic OAT, and further to examine whether levels of iron (Fe), copper (Cu), and selenium (Se) in seminal plasma can alter the associations. A total of 148 subjects (75 idiopathic OAT cases and 73 controls) were included in this study. Seminal plasma samples from all the participants were measured for levels of MDA, SOD, GSH, Fe, Cu, and Se. Unconditional logistic regression models were used to examine the associations between three oxidative stress indicators and the risk of idiopathic OAT. Bayesian kernel machine regression was performed to determine the joint effects of levels of three OS indicators on the risk of idiopathic OAT. Subgroup analyses were performed to explore whether the above associations can be different when Fe, Cu, and Se were in different levels. The level of MDA in seminal plasma was positively associated with the risk of idiopathic OAT, with adjusted odds ratio (OR) and 95% confidence interval (CI) of 2.38 (1.17, 4.83), and SOD and GSH levels were not associated with the risk of idiopathic OAT. In BKMR analyses, we found a significant positive association between the mixture of MDA, SOD, and GSH levels and the risk of idiopathic OAT at concentrations below the 65th percentile, while a negative association at concentrations above it. In subgroup analysis, a positive association was observed between MDA levels in seminal plasma and the risk of idiopathic OAT in the high-Cu group (adjusted OR = 3.66, 95%CI = 1.16, 11.57), while no significant association was found in the low-Cu group (adjusted OR = 1.43, 95%CI = 0.44, 4.58). Additionally, a negative association was found between GSH levels in seminal plasma and the risk of idiopathic OAT in the high-Se group (adjusted OR = 0.34, 95%CI = 0.11, 0.99), while no significant association was observed in the low-Se group (adjusted OR = 1.96, 95%CI = 0.46, 8.27). The levels of MDA, SOD, and GSH in seminal plasma were associated with the risk of idiopathic OAT, and the levels of Cu and Se in seminal plasma may alter the associations.

Metabonomic analysis of seminal plasma in necrozoospermia patients based on liquid chromatography-mass spectrometry.

In the pathological study of necrozoospermia-a form of sperm mortality-the underlying metabolic mechanism remains unclear. Thus, the aim of this study was to characterize metabolic alterations in the seminal plasma of necrozoospermic patients and to provide insights into the etiology of the disease. Seminal plasma samples were collected from patients diagnosed with necrozoospermia (n=28) as well as normozoospermia (n=37). The samples were analyzed using nontargeted metabolomics based on liquid chromatography-mass spectrometry (LC-MS). The raw data were subjected to multivariate analysis to identify metabolites correlated with necrozoospermia. Differential metabolites were subjected to pathway analysis using MetaboAnalyst. The results of the metabolomic analysis showed that there were 194 differential metabolites between the two groups; 129 metabolites were upregulated and 65 metabolites were downregulated. Among the differential metabolites, the top ten differential metabolites were choline, benzaldehyde, pyrazinamide, 5-aminoimidazole-4-carboxamide, and dihydrothymine. The following differential metabolite pathways were identified, and the top five metabolite pathways were arachidonic acid metabolism, steroid hormone biosynthesis, alanine aspartate and glutamate metabolism, bile secretion, and prostate cancer. The elevation of choline and 2-hydroxyglutarate levels in seminal plasma was an important finding, and the results also indicate that abnormalities in arachidonic acid metabolism and glutamate metabolism were an underlying pathological mechanism of necrozoospermia.

P-275 Oxidation reduction potential (ORP) levels in blood plasma could represent the ORP status in follicular fluid from oocyte donors and patients

Abstract Study question Does the evaluation of oxidation-reduction potential (ORP) levels in blood plasma from subfertile patients and oocyte donors represents the ORP status in follicular fluid? Summary answer ORP levels in blood plasma from oocyte donors and sub fertile patients could be an indicator of oxidative stress in follicular fluid What is known already Oxidation-reduction potential (ORP) measurement of seminal plasma is one method to diagnose oxidative stress in male patients, which can help physician to recommend antioxidants administration when it is elevated. Seminal plasma is easily accessible by masturbation. An empirical analogue for semen could be follicular fluid (FF). Unfortunately, it is very difficult to access to FF, unless a surgical procedure is performed. Hence, it is necessary to find indirect parameters to evaluate the ORP status in FF. Since blood plasma (BP) circulation provides antioxidants to the FF, we studied the BP ORP measurement to indirectly determine ORP levels in FF. Study design, size, duration Prospective study conducted at CITMER, Mexico from December 2022 to January 2023. We included under informed consent 15 oocyte donors (age 26.8 ± 4.06 years old) and 55 patients (age 35.2 ± 4.8 years old) undergoing IVF/ICSI. Participants/materials, setting, methods ORP levels in BP and FF from dominant follicles were measured at the same time of oocyte collection with MiOXSYS system. Oocytes were inseminated and zygotes were cultured until blastocysts stage. We calculated correlation of a) ORP levels in FF vs BP, b) ORP of FF and BP vs number of retrieved oocytes, c) ORP of FF and BP vs number of fertilized oocytes, d) ORP of FF and BP vs number of blastocysts Main results and the role of chance A total of 172 oocytes from oocyte donors were collected. After fertilization check, 135 zygotes (78%) were observed. A total number of 55 blastocysts (31%) were observed at day 5 + day 6. For patients, a total of 550 oocytes were collected, 360 zygotes were observed (65%) and 164 blastocysts at day 5 + day 6 were obtained (45%). The correlation between ORP levels from FF and BP for oocyte donors was 0.90 and 0.51 for patients. The overall ORP levels in FF and BP in patients were 136.2 ± 14.9 mV and 148.3 ± 15.97 mV, respectively. For oocyte donors, ORP levels in FF and BP were 160.3 ± 13.8 mV and 170.28 ± 17.67mV, respectively. There was a mild correlation of ORP levels in BP with numbers of oocytes retrieved (r = 0.25). The correlation of BP ORP with oocyte fertilization was 0.24 in patients with autologous oocyte utilization. The mean ORP levels of FF were 136.2 ± 14.9 mV, versus a mean ORP of 148.3 ± 15.97 mV in BP. The FF ORP levels between 100-150 mV [LRD1] were moderately correlated and predictive of blastocyst development [LRD2]. For donor oocytes there was a weak negative correlation [LRD3] of FF ORP with oocyte fertilization (-0.06) and a weak negative correlation for blastocyst formation was found (-0.18). Limitations, reasons for caution The MiOXSYS System ( Careous Biotechnology, Lituania) is a device calibrated to measure ORP levels in seminal plasma but not to measure ORP levels in FF nor BP. The number of analyzed data should be increased. Wider implications of the findings According to this study, the use of blood plasma to surrogate the measurements of ORP levels in follicular fluid could represent a further strategy to include female patients to be treated by antioxidants before an IVF treatment. Trial registration number None

Relationship among seminal antigenicity, antioxidant status and metabolically active sperm from Holstein-Friesian (Bos taurus) bulls

Sperm antigenicity has been implicated as a regulatory factor for acquiring fertilizing competence in the female reproductive tract. Overt immune response against the sperm proteins leads to idiopathic infertility. Hence, the aim of the study was to evaluate the influence of the auto-antigenic potential of sperm on the antioxidant status, metabolic activities and reactive oxygen species (ROS) in bovine. Semen from Holstein-Friesian bulls (n = 15) was collected and classified into higher (HA, n = 8) and lower (LA, n = 7) antigenic groups based on micro-titer agglutination assay. The neat semen was subjected to the evaluation of bacterial load, leukocyte count, 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay and lipid peroxidation (LPO) levels. Antioxidant activities in seminal plasma and intracellular ROS levels in the post-thawed sperm were estimated. The number of leukocytes was lower (p < .05) in the HA than the LA semen. The percentage of metabolically active sperm was higher (p < .05) in HA than the LA group. The activities of total non-enzymatic antioxidant, superoxide dismutase (SOD) and catalase (CAT) were higher (p < .05) while glutathione peroxidase activity was lower (p < .05) in the seminal plasma of LA group. The LPO levels of neat sperm and the percentage sperm positive for intracellular ROS in the cryopreserved sample were lower (p < .05) in the HA group. Auto-antigenic levels were positively correlated with the percentage of metabolically active sperm (r = 0.73, p < .01). However, the seminal auto-antigenicity was negatively (p < .05) correlated with the levels of SOD (r=–0.66), CAT (r=–0.72), LPO (r=–0.602) and intracellular ROS (r=–0.835). The findings were represented in graphical abstract. It is inferred that the higher auto-antigenic levels protect the quality of bovine semen by promoting sperm metabolism and lowering ROS and LPO levels.

Seminal plasma level of bisphenol A among males investigated for infertility in Benin City, Nigeria

Background: Evidence of an increasing rate of male infertility is replete in literature and has been attributed to several factors associated with environmental toxicants such as bisphenol A (BPA). It's presence in several products frequently used and the potential harmful effect on humans has raised serious concerns. Aims: The objective of this study was to determine the seminal plasma concentration of BPA among men investigated for infertility. Setting and Design: This was a cross-sectional study of men investigated for infertility in a tertiary hospital in Benin City, Nigeria. Materials and Methods: A total of 144 males (mean age: 40.55 ± 7.30) and 50 control subjects of proven fertility (mean age: 39.96 ± 5.68) were enrolled in the study. A structured questionnaire was used to obtain the sociodemographic data. Semen was collected after at least 3 days of sexual abstinence. Semen analysis was done according to the World Health Organization guidelines and seminal plasma BPA was determined using the enzyme-linked immunosorbent assay technique. Statistical Analysis Used: Chi-square, Student's t-test, and analysis of variance were used to compare measured variables. Pearson's correlation coefficient was used to determine the relationship between seminal plasma BPA and sperm indices. Results: Seminal plasma BPA was significantly higher (P &lt; 0.001) among infertile men than control subjects. The mean seminal plasma BPA concentrations increased with declining sperm count (P &lt; 0.001). Seminal plasma BPA levels strongly correlated inversely with sperm motility (r = −0.857, P &lt; 0.001) and sperm count (r = −0.821, P &lt; 0.001) and positively with abnormal morphology (r = 0.815, P &lt; 0.001). The likelihood ratios of elevated BPA concentration to cause harm to sperm count (14.562), motility (10.425), and morphology (27.838) were several folds higher among subjects being investigated for infertility. Conclusions: Seminal plasma BPA levels were significantly higher among infertile men than controls. The detection of high levels of BPA in the seminal plasma is an indication of human exposure to high concentrations of BPA, this information may be used to formulate environmental policy that may minimize human exposure and improve male reproduction.

Novel approaches on melatonin role: Presence of clock-hormone in fish seminal plasma

The study of melatonin is of great importance for the fundamental knowledge of any living system since it displays many different physiological roles, including being a potent natural antioxidant. To the best of our knowledge, there is no information regarding melatonin in fish seminal plasma. This study aimed to determine this clock-hormone levels in the seminal plasma of three aquaculture fish species: European seabass (Dicentrarchus labrax), gilthead seabream (Sparus aurata), and Senegalese sole (Solea senegalensis) (both wild and F1 breeders), and to explore melatonin potential role in fish reproduction and spermatozoa antioxidant status. Blood and seminal plasma were collected from fish during their reproductive season, at two different times of the day [mid-light (ML) and mid-dark (MD)], and melatonin concentration was determined by radioimmunoassay (RIA). Testosterone (T), 11-ketotestosterone (11KT), and total antioxidant status (TAS) were also determined, to investigate the putative role of seminal melatonin in fish reproduction, both at endocrine and antioxidant levels. For each species, Pearson's correlation analysis was performed between all possible factors. Blood plasma melatonin showed higher average values at night in the three species: gilthead seabream (808 ± 139 pg/mL), European seabass (364 ± 85 pg/mL), and Senegalese sole (248 ± 40 and 88 ± 11 pg/mL in F1 and wild males, respectively). However, melatonin levels in seminal plasma were species-specific: in European seabass, melatonin levels were not detectable at any time-point, whereas in gilthead seabream it was only found at MD (average of 21 pg/mL), and in Senegalese sole, different melatonin patterns were observed between F1 and wild males, but both had higher melatonin at MD (6.84 and 14.26 pg/mL, respectively). In gilthead seabream, at MD seminal melatonin levels correlated with the antioxidant status of seminal plasma. A relationship between blood melatonin and seminal TAS levels was observed in European seabass at ML: in this species, seminal melatonin could not be detected and the lowest seminal TAS levels were found. Regarding steroid analysis, opposite patterns in the seminal plasma of F1 and wild Senegalese sole were observed: at MD, wild Senegalese sole had substantially greater 11KT levels (2.53 ng/mL), whereas F1 males had higher T levels (1.92 ng/mL). In gilthead seabream, a positive correlation between T and ML blood melatonin and seminal TAS was observed. This study unraveled the species-specificity and daily changes of melatonin in fish seminal plasma.

Oxidative Stress in Seminal Plasma Negatively Influences Sperm Quality in Infertile Males

Objective: To investigate the association between malondialdehyde concentration in the seminal plasma of infertile men and sperm quality.Methods: This case-control study included 60 male participants ranging from 25-40 years old with half of them were fertile and the other half were infertile. Semen analysis was performed as per the WHO standards, and spectrophotometric measurement of seminal plasma malondialdehyde level was done.Results: Results showed that infertile men had significantly a higher mean level of malondialdehyde in their seminal plasma than fertile men (p&lt;0.001), which was inversely associated with sperm count and motility. Also, malondialdehyde was positively associated with abnormal sperm morphology.Conclusions: Elevated malondialdehyde levels in seminal plasma are associated with poor sperm quality. Malondialdehyde testing can, therefore, be used to diagnose and predict the outcome of male infertility. Antioxidants should also be administered to men with infertility to help counteract the effects of oxidative stress.

Determination of Parabens and Their Metabolites in Seminal Plasma from Chinese Men by Ultra High Performance Liquid Chromatography Tandem Mass Spectrometry (UPLC-MS/MS)

Parabens are endocrine-disrupting chemicals (EDCs) that have estrogen-like activities and may cause male reproductive disorders. Here, we developed a method for the simultaneous determination of four parabens (MeP, EtP, n-PrP, n-BuP) and two metabolites (4-HB and 3,4-DHB) in human seminal plasma by UPLC-MS/MS. The method was used to analyze 144 seminal plasma samples from Chinese males. MeP, EtP, n-PrP, and 4-HB were the dominant compounds. MeP, EtP, and n-PrP were significantly correlated to each other. In addition, 4-HB was significantly correlated to MeP, EtP, n-PrP, and 3,4-DHB, respectively. The results provide direct evidence that parabens and their metabolites are widely distributed in the male reproductive system. The study presents the paraben metabolites levels in human seminal plasma for the first time.

Nitric Oxide Level and Total Antioxidant Capacity in the Seminal Plasma of Infertile Smoking Men

Introduction&Aim of the work: to assess the effects of smoking on TAC and NO levels in seminal plasma of patients who suffer from oligoasthenozoospermia.

Seminal plasma protein profiles and testosterone levels as biomarker semen quality of candidate Madura bulls.

This study aims to determine the protein profile based on molecular weight (MW) and testosterone levels in seminal plasma (SP) that correlates to the semen quality of candidate Madura bulls. A total of 10 male candidate madura bulls underwent semen evaluation (motility, viability, membrane plasma integrity (MPI), and sperm concentration). The centrifuge was run at 1,200 rpm (4°C) for 20 min to collect SP. SP testosterone levels were measured using an Enzyme-linked immunosorbent assay. The characterization of SP proteins in Madura bulls was done using 1D sodium dodecyl-sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) analysis. All parameters were analyzed using Pearson correlation analysis. The results of the SDS-PAGE analysis found eight protein bands with the highest MW of 110 kDa and the lowest of 12 kDa. The mean and SD of SP testosterone levels were 20.58 ± 8.56 ng/ml, motility 59.32% ± 20.14%, viability 67.45% ± 20.22%, MPI 32.77% ± 16.52%, and sperm concentration 1,002.64 ± 429.33 106/mm3. Proteins with MWs of 110 and 91 kDa significantly correlated with MPI, and 110 kDa negatively correlated with sperm concentration (p < 0.05). Proteins with MWs of 73 and 36 kDa significantly correlated with SP testosterone levels, while proteins with MWs of 29 kDa significantly correlated with sperm viability (p < 0.05). The expressed protein fraction based on MW is closely related to the quality of semen, so it has the potential to be a biomarker of semen quality. Further research is needed to determine the specific proteins in certain fractions.

Influence of spirulina supplementation on growth performance, puberty traits, blood metabolites, testosterone concentrations, and semen quality in Barki male lambs.

The fertility and genetic value of the flock can be enhanced by selecting lambs with highly developed early puberty characteristics. Spirulina (SP) has been evaluated as a natural product supplement to boost lamb growth, immunity, and productivity. Study growth performance, blood metabolites, puberty development traits, semen quality, and seminal plasma biochemical concentrations of growing Barki lambs when supplemented with SP at different levels. in a 24 weeks study, 30 Barki male lambs weighing an average of 21.78 ± 2.56 kg, with a body condition score of 3.20 ± 0.55 and an age of about 16 ± 0.24 weeks were used. The lambs were randomly assigned to three groups (10 lambs each) of daily SP supplementation levels per lamb of 0 ml (control), 50 ml (SP1), and 100 ml (SP2). The SP powder was made into a water suspension using SP to water ratio of 1 g:10 ml. The growth characteristics, as well as the development of puberty, blood metabolites, and semen quality analysis of every lamb, were measured. The growth performance was greater (p < 0.05) in SP2 lambs compared with other lambs. While daily dry matter intake was not affected by SP treatment, feed efficiency had significantly improved in SP2 groups. Furthermore, the SP2 lambs have attained puberty at early ages than the control lambs. The testes volume of SP2 lambs was bigger (p < 0.05) than other groups throughout the pre-pubertal up to the puberty stage. The addition of SP had no effects on the total protein, glucose, and triglycerides concentrations. Meanwhile, the cholesterol concentration was lowest (p < 0.05) in the SP2 lambs. The blood and seminal plasma levels of alanine aminotransferase and aspartate aminotransferase decreased (p < 0.05) in the SP lambs more than their control counterparts. The levels of superoxide dismutase reduced glutathione, and total antioxidants had increased (p < 0.05) in the treated lambs compared with the control group. Further, the malondialdehyde levels decreased (p < 0.05) in the SP-treated lambs. Additionally, the SP2 lambs produced better semen quality than the control lambs. SP supplementation (100 ml/head/day) enhanced growth performance, feed efficiency, and antioxidative status, exerting a positive influence on the physiological parameters and sexual behavioral patterns at puberty in Barki lambs.

The Influence of blood, seminal plasma testosterone, growth hormone and cortisol levels on the sperm quality in merino rams

The research was conducted on 10 Merino rams to determine the effect of testosterone, growth hormone and cortisol levels on the sperm quality. A significant relationship was detected between blood and seminal plasma levels of testosterone (P ˂ 0.01). No GH on a measurable level was detected in the seminal plasma, and it was determined that plasma GH levels, just as testosterone levels began to increase with the approach of the reproductive season, and peaked at the start of the season. Plasma and seminal plasma cortisol levels, on the other hand, while increasing before the season, decreased significantly at the start of the season, and a remarkable correlation between plasma and seminal plasma levels was ascertained (P ˂ 0.01). An important negative relation which was observed between plasma GH levels and seminal plasma cortisol levels in Merino rams. It was found out that the hormones that were examined did not have a significant influence on sperm qualities, but there was an important negative relation only between the plasma GH levels and sperm volumes in the Merino (P ˂ 0.01). In conclusion, it may be stated that in rams, especially plasma testosterone and cortisol levels determine the plasma levels; both plasma and seminal plasma testosterone levels increase to a significant degree in the reproductive season.

Are There Associations between Seminal Plasma Advanced Oxidation Protein Products and Selected Redox-Associated Biochemical Parameters in Infertile Male Patients? A Preliminary Report.

Oxidative stress (OS) is one of the reasons for male infertility. Seminal plasma contains a multitude of enzymes and ions which influence OS and thus may affect male fertility. The aim of the study was to check for associations between seminal plasma advanced oxidation protein products (AOPP) concentrations and levels of selected biochemical parameters (total protein, iron, uric acid, magnesium, calcium) in infertile men, and establish whether they are associated with sperm disorders. Seminal plasma AOPP, as well as total protein, iron, uric acid, calcium, and magnesium concentrations, were determined for the following patient groups: normozoospermic (N; n = 33), teratozoospermic (T; n = 30), asthenoteratozoospermic (AT; n = 18), and oligoasthenoteratozoospermic (OAT; n = 28). AOPP concentrations were significantly higher in N and T groups in comparison to AT and OAT groups. Total protein concentrations were significantly lower in the T group in comparison to the AT and OAT groups, whereas iron concentrations significantly decreased in the OAT group in comparison to the T and N patients. AOPP differentiates AT patients from men with other sperm disorders. Our results suggest that asthenozoospermia may be connected with total protein levels. Insufficient iron levels may reflect a decrease in sperm count.

Curcumin Supplementation in Semen Extender Improves Post-Thaw Quality and Antioxidant Capacity of Gir Bull Spermatozoa

This investigation was carried out on semen of three Gir bulls at Cattle Breeding Farm, Junagadh (India) for a period of 6 months. The aim was to assess the effect of different concentrations of Curcumin, viz., 0 µM (Control), 5 µM, 10 µM, and 15 µM in Andromed extender on sperm quality along with oxidative stress parameters of cryopreserved semen. Semen ejaculates (n=30, 10/bull) with &gt;70% initial motility were included in the study. Soon after initial evaluation, the semen was extended in Andromed extender @ 80 million sperm per ml and was divided into four equal aliquots. The Curcumin stock solution (10 mM) was added in these aliquots to get a final concentration of 0, 5, 10, and 15 µM and sperm motility was checked. It was soon filled and sealed in medium French straws (0.5 ml), equilibrated at 4°C for 4 h and frozen in LN2 vapour using conventional wide mouth freezer. Thawing of straws was done next day in water bath at 37°C for 30 seconds, and various sperm quality parameters including oxidative markers were assessed. The results showed that supplementation of Curcumin to freezing extender improved semen quality and antioxidant capacity. The post-thaw sperm motility, sperm viability, HOST reactive sperm and acrosomal integrity in semen extended with 15 µM Curcumin concentration (57.33±0.67, 73.20±1.05, 64.77±1.56 and 72.43±1.38 %, respectively) were significantly (p&lt;0.05) higher and sperm abnormality (18.00±0.41%) was significantly (p&lt;0.05) lower as compared to those of control extender and the 5 µM Curcumin group. Mean malondialdehyde (MDA) level in post-thaw seminal plasma did not differ significantly (p&gt;0.05) among the groups, while total antioxidants activity (TAC) with 15 µM Curcumin (476.36±16.52 μmol/L) was significantly (p&lt;0.05) higher as compared to control and 5 µM Curcumin group. It is concluded that Curcumin @ 15 µM in semen extender improves post-thaw sperm quality and antioxidant capacity of cryopreserved Gir bull semen.

Cytokine profile in seminal fluid of infertile men with concomitant pathologies

The mechanisms of formation of male infertility are increasingly becoming immune dependent. This study aims to study the role of pro-inflammatory and anti-inflammatory cytokines in seminal fluid to clarify their role in spermatogenesis. This research is carried out at rheumatology and urology departments of Danylo Halytsky Lviv National Medical University (Ukraine). 45 infertile men aged 22–48 were examined. They were divided into 2 groups: first group – 22 men with a systemic autoimmune disease – rheumatoid arthritis; second group – 23 somatically healthy patients with idiopathic infertility. The control group included 27 fertile healthy men aged 22–48 years. Student's t-test was used to compare the significant difference in mean values between groups. Patients with autoimmune diseases (rheumatoid arthritis) had the highest rate of leukocytospermia, indicating a possible long-term inflammatory process. The TGF-β1 level was higher in men with idiopathic infertility, and in patients with autoimmune pathology was reduced compared to the control group, however these changes were not significant. At the same time in patient of both group the level a pro-inflammatory cytokine IL-18 increased approximately by 2.5 times. In patients with idiopathic infertility and patients with autoimmune pathology, a simultaneous increase in the IFN-γ level and a decrease in the IL-1β level were found. The ІL-6 level in seminal plasma of infertile men of both groups was increased. The ratio IL-10/TNF-α was decreased in seminal plasma of infertile men with rheumatoid arthritis. It was found that in autoimmune pathology, the increased level of IL-1β in blood serum compared to controls was associated with its reduced level in seminal fluid (r=-0.51, p&lt;0.05). In idiopathic infertility, a low level of IL-1β in seminal fluid was also associated with a low concentration of this cytokine in blood serum (r=0.62, p&lt;0.05). The functioning of the immune and reproductive systems of the male body is closely related and interdependent. Disorders of immune reactivity, which accompany the development of autoimmune pathology associated with disorders of reproductive function in men. The largest number of deviations of immune reactivity was found in infertile men with concomitant autoimmune diseases. Keywords: cytokines, male infertility, idiopathic infertility, rheumatoid arthritis.

Estimation of Serum and Seminal Plasma Levels of Glactin-1 in Non-Obstructive Azoospermia Cases and Their Correlations with the Rate of Sperm Retrieval: A Comparative Prospective Study

Remarkably, the current study is one of the first to deploy galectin-1 (Gal-1) in determining the degree of impairment of spermatogenesis among cases with non-obstructive azoospermia (NOA) as well as utilizing it as a biomarker to predict the rate of sperm retrieval in these patients. The purpose of the study was to evaluate the seminal plasma and serum levels of Gal-1 in NOA patients as well as their correlations with Johnsen's tubular biopsy scoring (JTBS). The current case control study included totally 48 patients with NOA whose ages ranged from 24 to 46 years old and 50 age matched healthy controls. Gal-1 levels were measured in both seminal plasma and serum of all subjects by the enzyme-linked immunosorbent assay (ELISA). A significant negative correlation between seminal plasma levels of Gal-1 and JTBS was detected (r= -0.281, p=0.048) in the NOA cases. Interestingly, the receiver operating characteristic (ROC) curve had demonstrated that the cutoff value of seminal plasma levels of Gal-1 in determining azoospermia was >0.735 ng/ml and the area under the curve (AUC) was 0.858. The sensitivity, specificity, positive predictive, and negative predictive values for seminal plasma levels of Gal-1 were 76, 92, 90.5, and 79.3, respectively. In addition, sensitivity, specificity, positive predictive, and negative predictive values for serum levels of Gal-1 were 38, 66, 52.8, and 51.6, respectively. Seminal plasma levels of Gal-1 are higher in NOA men versus healthy controls. Interestingly, negative correlation of seminal plasma levels of Gal-1 with JTBS was determined. Thus, it can be used as a good predictor for NOA cases.

An evidence of Humanin-like peptide and Humanin mediated cryosurvival of spermatozoa in buffalo bulls

Buffalo spermatozoa are vulnerable to cryo-injuries due to inherent deficiency of endogenous antioxidants, high polyunsaturated fatty acids (PUFA) content in plasma membrane and low cholesterol/phospholipid (C/P) ratio. Humanin is a potent cytoprotective agent that protects the cells against oxidative stress and apoptosis. The present study was designed to establish the presence of Humanin in buffalo and effect of Humanin supplementation on freezability of buffalo spermatozoa. Indirect immunofluorescence test revealed presence of Humanin in ejaculated and epididymal spermatozoa, and, elongated spermatids and interstitial space in the testicular tissue section. Humanin levels in seminal plasma were significantly and positively correlated with sperm concentration and individual progressive motility (IPM) in good (n = 22; IPM >70%) and poor (n = 10; IPM <50%) quality ejaculates. For supplementation studies, a total of 24 ejaculates (IPM ≥70%) were collected and each ejaculate was then divided into four aliquots. First aliquot was diluted with egg yolk-tris-glycerol (EYTG) extender without Humanin and served as control group (Group I). Rest three aliquots were diluted with extender containing 2 (Group II), 5 (Group III) and 10 μM Humanin (Group IV), respectively. Semen was cryopreserved using standard protocol and evaluated at pre-freeze for lipid peroxidation (LPO) and post-thaw stages for spermatozoa kinematics, LPO, mitochondrial membrane potential (MMP), capacitation, apoptotic status and DNA integrity. The treatment group that showed best results (5 μM) was compared with control group for in vitro fertility assessment by homologous zona binding assay. The LPO levels were lower (p < 0.05) in 5 and 10 μM Humanin supplemented group. The MMP and DNA integrity were higher (p < 0.05) in 5 μM group than other groups. F-pattern was higher (p < 0.05) and B-pattern was lower (p < 0.05) in 5 and 10 μM Humanin supplemented groups. Lower apoptotic and higher viable spermatozoa (p < 0.05) were observed in 5 μM Humanin group. The mean number of spermatozoa bound to zona pellucida was higher (p < 0.05) in 5 μM Humanin treated group than the control group. The study established the presence of Humanin in buffalo spermatozoa and seminal plasma for very first time and concluded that Humanin supplementation at 5 μM concentration improves the freezability and in vitro fertility of buffalo spermatozoa.