Seminal oxidation-reduction potential levels are not influenced by the presence of leucocytospermia.

Oxidative stress (OS) is characterised by an excessive amount of reactive oxygen species (ROS) which negatively affect sperm functions. In this study, the influence of leucocytes on seminal oxidation-reduction potential (ORP) and sperm DNA fragmentation (SDF) was investigated in 1,068 men. Seminal leucocyte concentration did not correlate with SDF, unadjusted ORP, ORP normalised for sperm concentration (sORP), ORP normalised for total motile sperm concentration (motORP) or total motile sperm count (TMSC-ORP). Although receiver operator characteristic (ROC) curve analyses show that leucocytospermia does not predict high sORP values (>1.34 mV/106 spermatozoa/ml), the motORP (AUC: 0.666) and TMSC-ORP (AUC: 0.683) predict the rate of leucocytospermia significantly (p = .0195 and p = .0085 respectively). Moreover, SDF can significantly predict leucocytospermia (AUC: 0.679; p = .011) and vice versa (AUC: 0.657, p = .0298). Our data confirm the association between OS and SDF. In conclusion, motORP and TMSC-ORP may be better predictive factors of leucocytospermia, probably because sperm motility, included in motORP and TMSC-ORP calculation, is the first seminal parameter to be affected by OS. Although all these parameters are indicative of OS, ORP values, SDF and leucocytospermia should be considered independently for the evaluation of redox seminal status, as they probe distinct seminal features.

Cytosolic phospholipase A2 and F2 isoprostanes are involved in semen quality and human infertility-A study on leucocytospermia, varicocele and idiopathic infertility.

Phospholipase A2 (PLA2 ) is involved in eicosanoid release, and F2 -isoprostanes (F2 -IsoPs), as free radical-generated eicosanoids released by PLA2 , are indicators of oxidative stress in different human conditions. This study investigated the interplay between cytosolic PLA2 (cPLA2 ), F2 -IsoPs and sperm features in male infertility, when the involvement of oxidative stress has been reported. Semen evaluation was performed following WHO guidelines, sperm ultrastructure was detected by transmission electron microscopy indicating a fertility index, and the percentages of sperm immaturity, apoptosis and necrosis. In sperm cells and seminal plasma, cPLA2 levels were determined by immunological method, whereas F2 -IsoPs by mass spectrometry. Sperm concentration, morphology, vitality and fertility index values were significantly lower in infertile groups compared with fertile men. An increase in sperm apoptosis and necrosis (p < .01), apoptosis (p < .01) and immaturity (p < .001) was detected in leucocytospermia, idiopathic infertility and varicocele, respectively. Seminal cPLA2 showed the highest value in varicocele group (p < .05), whereas seminal F2 -IsoPs increased in varicocele (p < .001) and leucocytospermia (p < .05) groups. In the whole population, F2 -IsoP and cPLA2 levels were positively correlated (p < .05). On the contrary, F2 -IsoPs and cPLA2 were not significantly different when investigated in sperm cells. Our data indicate that fatty acid oxidation/metabolism plays a role in different male reproductive pathological conditions.

Increase of leucocyte-derived extracellular traps (ETs) in semen samples from human acute epididymitis patients-a pilot study.

PURPOSE: To study the effector mechanism against pathogens of polymorphonuclear neutrophils (PMN) and macrophages, called ETosis, involving the release of extracellular traps (ETs) in patients with acute epididymitis. To assess the different ET phenotypes present in semen samples and to identify correlations between ETosis and clinical parameters. MATERIALS AND METHODS: Samples from patients diagnosed with acute epididymitis were examined and compared with samples from uninfected controls. Biochemical analyses of seminal fluid included determination of peroxidase, α-glucosidase, fructose, and elastase levels. ETosis in semen was determined through presence of citrullinated histones, global histones, and extracellular DNA. Different ETosis phenotypes such as spread ETs, aggregated ETs, and diffuse ETs were identified by co-localisation of extruded DNA with myeloperoxidase and global histones. Anti-CD15+ and anti-CD68+ antibodies were used to identify different cell lines. RESULTS: Revealed a high number of ETs compared with the control group. The mean number of CD15+PMN and CD68+ macrophages was higher in the acute epididymitis group. ETosis increase in ejaculates correlated with clinical parameters such as enhancement of elastase concentrations and diminution of fructose in the semen. CONCLUSIONS: This work shows for the first time the presence of ETs and their components in semen from patients with acute epididymitis. The presence of infections is an important factor for induction of ETs in semen. Furthermore, the presence of ETosis in ejaculates is suggestive of developing infectious processes and might possibly have a diagnostic value.

Therapeutic options in male genital tract inflammation.

Chronic inflammatory conditions of the genital tract are still unsatisfactorily recognised in the workup of male infertility due to inappropriate definitions and inconsistent diagnostic criteria. The most popular term used for description of both, infections and inflammation in the genital tract is MAGI (male accessory gland infection). In asymptomatic patients, the diagnosis is primarily based on leucocytospermia (i.e., more than 1 million peroxidase-positive leucocytes per ml ejaculate), although ongoing infections should be identified and distinguished from post-infectious or non-infectious inflammatory disease. In addition to alterations of the basic semen parameters, sperm functions -and DNA integrity may be affected by chronic inflammation of the male genital tract. Despite considerable diagnostic drawbacks and a rather limited database concerning evidence-based therapy, adequate management of affected patients appears mandatory. Antibiotic treatment aims at the eradication or reduction of pathogenic bacteria in the ejaculate. Available studies suggest, that NSAID are effective in chronic inflammatory conditions. Moreover, low-dose corticosteroids, mast cell blockers, and other immune-modulatory compounds as well as a sequential adjuvant treatment with antioxidants can be considered as therapeutic options.

Sexually transmitted agents and their association with leucocytospermia in infertility clinic patients.

In this study, the presence of Chlamydia trachomatis, Neisseria gonorrhoeae, Trichomonas vaginalis, Mycoplasma hominis, Mycoplasma genitalium, Ureaplasma urealyticum and Ureaplasma parvumDNA were investigated using real-time multiplex PCR detection kit in the urine samples of 50 patients who presented to the infertility polyclinic. Patients were classified into two groups in accordance with the WHO leucocytospermia criteria as over 1 × 106 /ml (n = 25) and below 1 × 106 /ml (controls; n = 25). In addition, the semen sample results of the patients were evaluated. The mean leucocyte count in the leucocytospermia group was detected as 3.4 × 106 /ml. Agent positivity was found in 12 of 25 patients in the leucocytospermia group (U. parvum 10, U. urealyticum 3, M. hominis 3) and 9 of 25 patients in the control group (U. parvum 7, U. urealyticum 2, M. hominis 1). A chi-square test evaluation revealed no statistically significant difference between groups. (p = 0.390). The statistical evaluation using the Mann-Whitney U and chi-square tests revealed no statistically significant negative effect of agent positivity on semen analysis parameters in either group (p > 0.05). Although no significant difference was detected between the groups, this study provides data about the prevalence of asymptomatic sexually transmitted diseases in patients presenting to the infertility polyclinic.

In vitro antioxidant effect of curcumin on human sperm quality in leucocytospermia.

Decreased sperm quality was caused by oxidative stress in semen from patients with leucocytospermia. Curcumin is a traditional Chinese herbal monomer extracted from Zingiberaceae turmeric and zedoary turmeric and has antioxidative and anti-inflammatory effects. This study aimed to investigate the effects and specific molecular mechanisms of curcumin on sperm quality in patients diagnosed with leucocytospermia. Forty cases of semen samples were collected from patients with leucocytospermia and 35 cases from normal fertile male. Computer-assisted semen analysis (CASA) was used to detect sperm motility after curcumin incubation. ELISA was used to measure the changes in H2 O2 , sperm mitochondrial DNA (mtDNA), cytochrome B (Cyt B) and NADH dehydrogenase 5 (NADH5) contents before and after curcumin treatment. The results indicate that curcumin can significantly improve sperm motility from the patients with leucocytospermia. After curcumin treatment, the level of the H2 O2 was significantly decreased in the supernatant of curcumin-incubated spermatozoa from leucocytospermic patients. The content of mtDNA was significantly decreased, while the content of Cyt B and NADH5 in spermatozoa was significantly increased. In conclusion, curcumin can significantly improve sperm motility of leucocytospermic patients, against the oxidative damage induced by H2 O2 . Therefore, curcumin may play a role in mitigating the H2 O2 -induced injury to sperm.

Seminal Corynebacterium strains in infertile men with and without leucocytospermia.

This study aimed to identify seminal Corynebacterium strains in infertile men with and without leucocytospermia. Semen samples from 60 infertile men were allocated into two equal groups: semen samples with leucocytospermia and semen samples without leucocytospermia. Semen culture for Corynebacterium species was carried out on Columbia agar medium confirmed by Gram-stained film and biochemical tests followed by analytical profile index biotyping and antibiotic susceptibility. Bacterial isolates were detected in 20/60 semen cultures (33.3%) as Corynebacteria, Staphylococci, Alpha haemolytic streptococci and E. coli. In all, 12/60 (20%) had Corynebacterium positive semen culture, whereas C. seminal was the major isolated species followed by C. amycolatum, C. jekium and C. urealyticum. There was nonsignificant difference between patients with/without Corynebacterium positive culture regarding sperm concentration and normal sperm morphology; however, in positive cultures sperm motility was significantly lower compared with negative cultures. Antimicrobial sensitivity among Corynebacteria strains was highest for vancomycin, rifampicin then imipenem, ampicillin + sulbactam, ciprofloxacin. It is concluded that positive semen cultures for different Corynebacteria species were demonstrated in infertile men, whereas Corynebacterium seminale was the most common isolated species. Vancomycin, rifampicin then imipenem and ampicillin + sulbactam are recommended as sensitive antibiotics.

Seminal plasma leptin and spermatozoon apoptosis in patients with varicocele and leucocytospermia.

Excessive apoptotic spermatozoon death is associated with male infertility. Leptin regulates apoptosis in several cell types. We prospectively investigated if seminal plasma leptin mediates spermatozoon apoptosis in 74 varicocele (VC) patients and 70 leucocytospermia patients. Spermatozoa from 40 normospermic men were used as controls. Routine semen analysis, spermatozoon apoptosis rate, seminal plasma leptin, reactive oxygen species (ROS) and tumour necrosis factor-alpha (TNF-α) levels were measured. In VC and leucocytospermia patients, seminal plasma leptin levels and spermatozoon apoptosis rates were significantly higher compared with controls. In the VC group, seminal plasma ROS levels were significantly higher compared with controls; there were no significant differences in TNF-α levels. In the leucocytospermia group, both ROS and TNF-α levels were significantly higher compared with controls. In both the VC and leucocytospermia groups, there was a significant positive correlation between the spermatozoon apoptosis rate and leptin levels and ROS and leptin levels. There was a significant correlation between leptin and TNF-α levels in the leucocytospermia group. Seminal plasma leptin levels correlate significantly with spermatozoon apoptosis rate, and leptin may be a spermatozoon pro-apoptotic factors. The generation of ROS is a possible mechanism. Leptin may induce apoptosis via TNF-α in leucocytospermia patients.

Macrophage metalloelastase-12 is detectable in human seminal plasma and represents a predictor for inflammatory processes in the male genital tract.

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

Association of IgA secretory component sialylation with leucocytospermia of infertile men - a pilot study.

The aim of our pilot study was to check whether the differences in IgA secretory component (SC) sialylation are associated with leucocytospermia. In normozoospermic and leucocytospermic seminal plasmas, 78-kDa and 63-kDa SC immunoreactive bands were observed. The SC sialylation was analysed by lectin blotting, using sialo-specific lectins MAA (Maackia amurensis agglutinin) and SNA (Sambucus nigra agglutinin). Specific reactivity of 63-kDa SC with MAA and SNA was higher than 78-kDa SC in both analysed seminal groups. The analysis of seminal SC sialylation might be a valuable diagnosis tools for the evaluation of fertility problems related with leucocytospermia.

A Successful Pregnancy Following Intracytoplasmic Sperm Injection in a Breast Cancer Survivor: A Case Report.

This report documents the case of a 36-year-old female diagnosed with stage I invasive ductal carcinoma of the left breast who, alongside her 39-year-old husband, sought fertility assistance at our center due to primary infertility. Having survived cancer twice in the span of their seven-year marriage, the couple faced the challenge of overcoming both the repercussions of cancer treatment and difficulties in conceiving. Initial attempts through three intrauterine insemination (IUI) cycles proved unsuccessful, leading the couple to opt for in vitro fertilization (IVF). The fertility assessment of the husband revealed the presence of several pus cells and a high sperm DNA fragmentation index (DFI). To address this, a medication regimen was administered to improve sperm quality. Concurrently, the female underwent controlled ovarian stimulation (COS) with the anti-estrogen agent letrozole to mitigate the risk of estrogen surges that could compromise her health. Subsequently, oocytes were retrieved from the female, and intracytoplasmic sperm injection (ICSI) was used to facilitate fertilization with her husband's sperm. Following successful embryo development, the patient underwent embryo transfer (ET), resulting in a positive beta-human chorionic gonadotropin (beta-hCG) result, signifying a successful conception. This case report highlights the intricate challenges faced by individuals with a history of breast cancer, emphasizing the delicate balance required in managing infertility in such circumstances. The described approach, involving personalized treatments and meticulous care, underscores the possibility of achieving successful conception for females struggling with fertility issues post-cancer survival. The documented journey serves as a testament to the resilience of individuals facing the dual challenges of cancer survival and infertility, offering insights into the complexities of their reproductive healthcare.

Papanicolaou-Stained Cytosmear Preparations in the Evaluation of Leucocytospermia: A Tertiary Centre Experience and Assessment of Utility.

Objectives Leucocytospermia is a rare cause of infertility with a variable incidence in infertile men. In many andrology laboratories, semen analysis is primarily centered on analyzing basic parameters (sperm count, motility, and viability). We examined the role of cytomorphological analysis on Papanicolaou (PAP)-stained smears in the work up of male infertility, with special reference to leukocytospermia and assessed the morphological features of sperms in these cases. Materials and Methods All cases signed out as "leukocytospermia" between November 2017 and January 2018 were evaluated. Cases showing pus cells (≥ 1/5hpf) on cytosmear evaluation were also analyzed. Parameters obtained on modified Neubauer's chamber, wet mount preparations, and autoanalyzer SQA-IIC-P (Medical Electronic Systems, Israel) were tabulated. PAP-stained smears were examined to assess morphological defects and other findings, if any. Results Out of 348 semen samples, 6 (1.72%) were diagnosed as leukocytospermia, all displaying pus cells on cytomorphological evaluation. Five cases having white blood cells < 1 × 10 9 /L were analyzed, which displayed the presence of pus cells (≥ 1/5 hpf) on cytosmear preparations (1.43%). Nine cases (81.8%) showed reduced motile sperm concentration, functional sperm concentration, and sperm motility index. Morphological evaluation of the PAP-stained smears showed mid-piece defects in 18.1% cases. Conclusion Diligent cytomorphological evaluation of semen is essential to diagnose leukocytospermia, assess morphological defects, and serve as a quality control measure.