Sperm Vitality Research Articles

Does folate deficiency alter sperm parameters in infertile males?

Introduction: There is not enough evidence regarding the influence of folate deficiency in the etiology of idiopathic male infertility. Objectives: The objective of the current study was to examine whether there is an association between serum levels of folate and sperm parameters among infertile males. Patients and Methods: This cross-sectional study was undertaken on a sample of infertile males referring to an infertility treatment clinic in Isfahan province, Iran. Semen parameters, comprising sperm concentration, progressive/total motility, morphology, semen leukocyte, agglutination, and vitality were assessed based on standard protocols. The sperm DNA fragmentation assay (SDFA) and sperm chromatin maturity assay (SCMA) were also assessed by the sperm chromatin dispersion and aniline-blue tests respectively. Results: In general, 70 infertile males including 46 with sufficient serum folate level and 24 with insufficient serum folate level were examined. Subjects with sufficient serum folate level had a significantly higher mean of sperm concentration, vitality, progressive motility, total motility, SDFA, and SCMA (P<0.05). Higher serum folate level was also associated with lower risk of abnormal sperm agglutination (OR: 0.09, 95% CI: 0.01-0.65), and morphology (OR: 0.12, 95% CI: (0.02-0.60) after adjustment for potential confounders. Conclusion: The present study found a significant association between serum folate level and infertility among Iranian infertile males. However, prospective large-scale population-based research is required to confirm the finding.

Effect of obstructive sleep apnea on semen quality.

Obstructive sleep apnea (OSA) has several notable complications such as hypertension and diabetes. Studies have also shown that OSA is associated with erectile dysfunction and reduced androgen levels. However, the effect of OSA on semen quality remains poorly studied. Men attending a tertiary reproductive center for semen analysis were tested with a portable sleep breathing monitor. Patients were divided into four groups based on their apnea hypopnea index: none, mild, moderate, and severe obstructive sleep apnea. Differences between groups were assessed using χ2, and associations were tested with multiple regression analysis. We included a total of 175 male subjects with a mean age of 32.2 ± 3.6years. There were significant differences between groups in progressive sperm motility (%) (43 ± 16, 42 ± 17, 36 ± 18, 29 ± 18, respectively; p = 0.002), total motility (%) (59 ± 19, 59 ± 20, 49 ± 21, 42 ± 20, respectively; p = 0.010), and vitality (%) (80 ± 10, 81 ± 11, 79 ± 8, 72 ± 19, respectively; p = 0.039). Asthenospermia (progressive motility < 35%) was significantly more common in subjects with OSA (χ2 = 5.195, p = 0.023). In multiple regression models, after adjusting for age and body mass index, apnea hypopnea index remained negatively and significantly associated with progressive motility, total motility, and vitality. OSA is an independent risk factor for sperm motility and vitality, and further investigation is now needed to determine if continuous positive pressure ventilation or other therapies can improve semen quality in these patients.

Eosin Nigrosin staining technique in assessment of sperm vitality in medical laboratories – A snippet from our experience on implementing the staining, interpretation and quality control procedures

Eosin Nigrosin staining for assessment of sperm vitality is an essential component of basic semen analysis as it helps differentiate between dead and immotile sperms, and has clinical implications in terms of patient treatment and follow up. This staining technique involves minimal use of reagents and simple procedural steps. Standardization of the same is pertinent to warrant accurate and reproducible results in medical laboratories, even those not specialized in infertility care. We wish to share our hands on experience through various stages in implementing this staining technique from challenges faced to putting quality control processes in place as reference for peers.

Effect of systemic isotretinoin therapy on semen parameters

Purpose Vitamin A has multiple functions in the human body, being involved in growth, epithelial differentiation, vision, immune function and reproduction. While normal spermatogenesis is influenced by several factors, it requires vitamin A. Systemic isotretinoin is a vitamin A derivative that is used in the treatment of many dermatological diseases, especially acne vulgaris (AV). There is limited research on the changes in semen parameters after systemic isotretinoin therapy in humans. Our study investigates the presence of varicoceles in patients undergoing systemic isotretinoin therapy for AV and examines whether there were any changes in the semen parameters before and after treatment. Methods Included in the study were 46 men patients who were scheduled for systemic isotretinoin therapy for AV. Before treatment, the patients underwent a physical examination and ultrasonography for varicoceles assessment. The patients underwent spermiogram before treatment and after 6 months of treatment. The spermiogram assessments included semen volume, sperm concentration, total sperm count, progressive motility, viability and sperm morphology. Results After treatment, there was an increase in semen volume, sperm concentration, total sperm count, progressive motility and vitality from the pre-treatment values, but a deterioration in the sperm morphology (p < .05). Comparing patients with and without varicoceles revealed more changes in semen parameters after treatment in those with varicoceles. There was a statistically significant difference in sperm concentration (p < .001). Conclusions Systemic isotretinoin therapy negatively affects sperm morphology, but has positive effect on other semen parameters, and these changes in semen parameters occur more frequently in patients with varicoceles. KEY MESSAGES Acne vulgaris is a very common disease and systemic isotretinoin is used as the most effective agent in its treatment. Systemic isotretinoin positively affects semen parameters except sperm morphology. Changes in semen parameters are more common in patients with varicocele.

Effects of melatonin on a d-galactose-induced male reproductive aging mouse model

Understanding the aging mechanism of the male reproductive system and developing anti-aging interventions are essential for preventing age-related male infertility. The pineal hormone melatonin has been effectively used as an antioxidant and anti-apoptotic molecule in various cells and tissues. However, the effects of melatonin on d-galactose (D-gal)-induced aging have not been studied with regards to testicular function. Thus, we investigated whether melatonin suppresses the dysfunction of male reproductive function induced by D-gal treatment. The mice were divided into the following four groups receiving treatments for six weeks: phosphate-buffered saline (PBS) group, d-galactose (200 mg/kg) group, melatonin (20 mg/kg) group, and d-galactose (200 mg/kg)+ melatonin (20 mg/kg) group. At six weeks of treatments, sperm parameters, body and testes weight, gene and protein expression of germ cell and spermatozoa marker were analyzed. Our results showed that melatonin suppressed the decrease in body weight, sperm vitality, motility, and gene expression levels of spermatozoa markers such as Protamine 1, PGK2, Camk4, TP1, and Crem in the testis of D-gal-induced aging models. However, the gene expression levels of the pre-meiotic and meiotic markers in the testes did not change in the D-gal-injected model. The injection of D-gal impaired the decreased expression of steroidogenic enzyme genes, such as HSD3b1, Cyp17a1, and Cyp11a1, but melatonin inhibited the decrease in the expression of these genes. In addition, protein levels of spermatozoa and germ cell markers were evaluated by immunostaining and immunoblotting. Consistent with the qPCR results, PGK2 protein levels were decreased by d-galactose treatment. A decrease in PGK2 protein levels by D-gal was inhibited by melatonin treatment. In conclusion, melatonin administration improves testicular function with age.

Melatonin alleviates cisplatin-induced mice spermatogenesis defects

Cisplatin (CDDP) is a chemotherapeutic drug that is used to treat many different types of tumors. However, it also has significant adverse effects on male reproduction, which are partially mediated oxidative damage. Melatonin (MLT) is a promising antioxidant that can be used for reproductive protection. In this paper, we investigated the effect of CDDP on spermatogenesis, as well as MLT’s potential role in reproductive protection. CDDP (5 mg/kg BW) significantly reduced male mice testosterone levels and decreased sperm vitality and progressive motility. Additionally, a lower percentage of stage VII and VIII seminiferous tubules were observed in CDDP-treated mice. MLT administration significantly alleviated CDDP-induced testicular damages, CDDP-induced lowered male fertility in vivo, and enhanced in vitro embryonic development of two cells and blastocysts. These changes may be due to CDDP-mediated spermatogenesis defects in germ cell and Leydig cell proliferation, which are reflected in abnormal PCNA, SYCP3, and CYP11A1 expression levels and can be improved by MLT. CDDP treatment significantly decreased the total antioxidant capacity (TAC), as well as SOD and GSH levels, and increased MDA levels in mice testis, leading to increased apoptosis of germ cells and increased BAX/BCL2 ratios in mice testis. MLT treatment may reduce germ cell apoptosis by reducing oxidative damage in mice testis. This study demonstrated that CDDP affects sperm fertility by altering germ cell and Leydig cell proliferation via increased oxidative damage and that MLT can attenuate these damages. Our work provides potential information for further research on the toxic effects of CDDP and the protective effects of MLT on male reproduction.

Mitochondria-Stimulating and Antioxidant Effects of Slovak Propolis Varieties on Bovine Spermatozoa

Propolis is a natural honeybee product, which is known for its beneficial pharmacological effects and use in traditional medicine. This study aspired to investigate the chemical and antioxidant properties of five propolis specimens, followed by the analysis of their in vitro impact on bovine spermatozoa vitality and oxidative profile. Semen samples from 10 breeding bulls were incubated in the absence or presence of 50 µg/mL propolis extracts, and selected sperm quality parameters including motility, mitochondrial activity, cAMP concentration, NAD+/NADH ratio, reactive oxygen species (ROS) and superoxide production were assessed at 2 h and 24 h. Sperm motion behavior and mitochondrial function were stabilized particularly in the presence of propolis collected from Hrabské and Litpovský Hrádok, Slovakia. At the same time, all propolis specimens acted as significant ROS quenchers and stabilized the oxidative milieu of the sperm cultures. Our data suggest that propolis may stabilize the mitochondrial function of spermatozoa while preventing excessive oxidative insults, thereby showing the potential to sustain the sperm motility of extended semen.

The Effects of Medium-Term Intake of Hydrogen-Rich Water on Sperm Quality Biomarkers in Normospermic and Oligospermic Men: A Randomized Controlled Pilot Trial

The main aim of this randomized controlled pilot trial was to evaluate the effects of hydrogen-rich water on spermiogram parameters in normospermic and oligospermic men. As many as 12 healthy young men (age 29.1 ± 5.9 years; n = 6 normospermic; n = 6 oligospermic) volunteered. Participants were allocated in a double-blind manner to receive 1 L of hydrogen-rich water per day or 1 L of tap water fortified with magnesium for 8 weeks. Following hydrogen-rich water supplementation, sperm concentration and morphology tended to increase by 12.4 million per milliliter (95% CI; –31.8 to 56.6), and live cells increased by 3.8% (95% CI; –12.5 to 20.1), respectively (P ≤ 0.30). A significant difference between the two interventions was found in sperm vitality (P = 0.03), with hydrogen-rich water being superior to the placebo in terms of an increase in the number of live sperm cells. Subgroup analyses revealed no significant differences between interventions in sperm viability outcomes in oligospermic men (P &gt; 0.05). However, sperm count increased by over 20 million in two of the three oligospermic men (66.7%) receiving hydrogen-rich water, while no oligospermic men in the placebo group increased sperm concentration to euspermic levels (P &lt; 0.01). These preliminary findings suggest hydrogen-rich water is an effective intervention for improving some aspects of male subfertility; further large-sample trials are warranted to corroborate our results.

Levels of DNA, Protein, Lipid Oxidation and Apoptosis Biomarkers in Semen of Men with Hyperviscous Semen: A Cross-Sectional Study.

Semen hyperviscosity is a threatening cause of abnormal spermatozoa and infertility in men. We aimed to evaluate oxidative stress, antioxidants depletion and sperm apoptosis as main reasons for poor quality of spermatozoa in men with hyperviscous semen. In this cross-sectional study, ejaculate specimens were collected from fertile (n=102) and infertile men with hyperviscous semen (n=123) and without semen hyperviscosity (n=143). Total antioxidant capacity (TAC), glutathione (GSH), malondialdehyde (MDA), protein carbonyl (PC), 8-hydroxydeoxyguanosine (8-OHdG), and were measured in semen samples to estimate oxidative stress status. Gene expression pattern of BAX, CASPASE-9, CASPASE-3, and BCL2 was assessed to estimate sperm apoptosis. The average of sperm count, normal morphology, normal motility, and sperm vitality in men with hyperviscous semen was significantly lower than infertile subjects without hyperviscous semen (P<0.01). Men with hyperviscous semen exhibited higher levels of PC (8.34 ± 1.03 nmol/mg vs. 6.01 ± 0.93 nmol/mg, P=0.008), MDA (1.14 ± 0.27 nmol/ ml vs. 0.89 ± 0.22 nmol/ml, P=0.031), 8-OHdG (259.71 ± 24.59 ng/ml vs. 197.13 ± 18.47 ng/ml, P=0.009), but lower TAC contents (1250.44 ± 66.23 μM/L vs. 1784.31 ± 89.87 μM/L, P=0.018) and GSH (3.82 ± 1.05 μM vs. 5.89 ± 0.87 μM, P=0.021) than men with non-viscous semen. The expression of BAX, CASPASE-3 and CASPASE-9 genes in men with hyperviscous semen was significantly increased by 1.39-fold (P=0.041), 1.47-fold (P=0.046), 1.29-fold (P=0.048), respectively, as compared with the infertile subjects without hyperviscous semen. However, BCL2 expression in infertile men without hyperviscous semen was higher compared to those with hyperviscous semen (1.36-fold, P=0.044). Hyperviscous semen is markedly associated with depletion of seminal plasma antioxidants, sperm membrane lipid peroxidation, DNA and protein oxidation, and sperm apoptosis. Antioxidant therapy might be considered as a valuable strategy to protect sperm cells against oxidative damage in cases with seminal fluid hyperviscosity.

MP30-01 TEMPORAL TRENDS IN SPERM COUNT OF FERTILE PATIENTS: A SYSTEMATIC REVIEW AND META-REGRESSION ANALYSIS

You have accessJournal of UrologyCME1 Apr 2023MP30-01 TEMPORAL TRENDS IN SPERM COUNT OF FERTILE PATIENTS: A SYSTEMATIC REVIEW AND META-REGRESSION ANALYSIS Rossella Cannarella, Raneen Sawaid Kaiyal, Shinnosuke Kuroda, Kieran Lewis, Sarah Vij, and Scott Lundy Rossella CannarellaRossella Cannarella More articles by this author , Raneen Sawaid KaiyalRaneen Sawaid Kaiyal More articles by this author , Shinnosuke KurodaShinnosuke Kuroda More articles by this author , Kieran LewisKieran Lewis More articles by this author , Sarah VijSarah Vij More articles by this author , and Scott LundyScott Lundy More articles by this author View All Author Informationhttps://doi.org/10.1097/JU.0000000000003258.01AboutPDF ToolsAdd to favoritesDownload CitationsTrack CitationsPermissionsReprints ShareFacebookLinked InTwitterEmail Abstract INTRODUCTION AND OBJECTIVE: While current evidence suggests the existence of a downward trend of sperm concentration and count in unselected men from both industrialized and underdeveloped countries, these data have been controversial due to the heterogeneity of the study populations and the differences in World Organization Health (WHO) manual edition used. In response, we performed a focused meta-analysis specifically in men with proven paternity and restricted our analysis to studies using the 5th edition of the WHO manual (2011-2020). METHODS: A literature search was performed using Scopus, PubMed, Ovid, Embase and Cochrane databases according to PRISMA guidelines. Men were defined as fertile if demonstrating proven paternity with variable interval between conception and semen analysis. The primary outcomes of this study was sperm concentration, and the secondary outcomes included semen volume, progressive motility, total sperm count, and morphology. A meta-regression analysis was performed to determine the change in the selected outcomes over time. RESULTS: 4286 abstracts were initially reviewed, 139 full text articles were fully assessed for eligibility, and a total of 8 articles were included in the final cohort. Sperm concentration (mean 86.2M/ml, 95% CI 49.0, 123.4, I2= 99.1%; p<0.001) did not change from 2011 to 2020 (p= 0.4105). Only three studies reported total sperm count (322.7M/ejaculate, 95% CI 52.2, 593.2, I2= 99.9%; p<0.001), and the data were insufficient to run a meta-regression analysis. Semen volume (3.6 ml, 95% CI 2.5, 4.7, I2= 99.3%; p<0.001) did increase from 2011 to 2020 (+0.303 mL/year; p<0.01). Interestingly, both sperm motility (59.7%, 95% CI 53.5, 66.0, I2= 97.8%; p<0.001) and progressive motility (48.5 %, 95% CI 47.6, 49.3, I2= 95.4%; p<0.001) significantly reduced over the study time period (-2.2%/year, p<0.0001 and -3.1%/year, p<0.01). Sperm morphology (71.8 %, 95% CI 71.0, 71.6, I2= 99.7%; p<0.001) and vitality (30.4 %, 95% CI 19.6, 41.2, I2= 97.1%; p<0.001) did not change from 2011 to 2020. CONCLUSIONS: The results of this study suggest that sperm motility, but not total count, may be falling for unknown reasons. Further work is needed to better understand the mechanisms and clinical implications for this finding. Source of Funding: None © 2023 by American Urological Association Education and Research, Inc.FiguresReferencesRelatedDetails Volume 209Issue Supplement 4April 2023Page: e391 Advertisement Copyright & Permissions© 2023 by American Urological Association Education and Research, Inc.MetricsAuthor Information Rossella Cannarella More articles by this author Raneen Sawaid Kaiyal More articles by this author Shinnosuke Kuroda More articles by this author Kieran Lewis More articles by this author Sarah Vij More articles by this author Scott Lundy More articles by this author Expand All Advertisement PDF downloadLoading ...

Analysis of sperm separation protocols for isolating cryopreserved human spermatozoa.

Sperm cryopreservation is a valuable tool for the long-term preservation of male fertility. Thus, determining the optimal technique for isolating spermatozoa post-thaw is vital to ensure recovery of the highest quality spermatozoa with minimal iatrogenic damage. This not only enhances the chances of successful conception but also reduces the risk of genetic damage in the embryo. To address this issue, human semen samples were cryopreserved using a slow freezing protocol and Quinn's Advantage™ Sperm Freeze medium. The samples were subsequently thawed and subjected to three types of sperm isolation procedures: direct swim-up, density gradient centrifugation, and electrophoretic separation using the Felix™ device. Cryopreservation led to the anticipated loss of sperm motility and vitality in association with increases in lipid peroxidation and DNA damage. Following sperm selection, all three isolation techniques resulted in an increase in sperm motility which was particularly evident with the swim-up and Felix™ procedures. The latter also significantly improved sperm vitality. There were no differences between sperm separation techniques with respect to morphology, and mitochondrial reactive oxygen species generation remained essentially unchanged when cell vitality was taken into account. By contrast, major differences were observed in DNA integrity and lipid aldehyde formation, where Felix™ isolated cells exhibiting significantly less DNA damage than the other isolation procedures as well as lower levels of 4-hydroxynonenal formation. Electrophoretic sperm isolation, therefore, offers significant advantages over alternative separation strategies, in terms of the quality of the gametes isolated and the time taken to achieve the isolation. Long-term storage of sperm is vital to assisted reproductive technology because it permits the preservation of fertility that might be compromised as a result of factors such as chemotherapy or vasectomy. This goal can be achieved via cryopreservation - the freezing of cells to -196°C. When the sperm are subsequently required for conception, they must be carefully separated from the cryopreservation medium in a manner that maximizes the chances of successful conception and minimizes the risk of genetic defects in the offspring. In this paper, three isolation techniques were compared for their ability to separate ideal sperm from semen and media following cryopreservation. It was found that cryopreservation led to lower levels of motility and vitality and created higher levels of DNA and cell membrane damage. Of the three techniques compared, only cells separated on the basis of their size and electric charge (electrophoretic isolation) exhibited significantly lower levels of DNA fragmentation.

Antichlamydia antibodies and sperm quality among male partners of infertile couples ins Nigeria.

The impact of Chlamydia trachomatis on semen quality has been studied with varied results. To determine the prevalence of antichlamydial antibodies and their relationship with sperm quality among male partners of infertile couples in Enugu, South-East Nigeria. It was a cross-sectional study of infertile male partners of couples attending infertility clinics at the University of Nigeria Teaching Hospital (UNTH) Ituku-Ozalla, Enugu, Nigeria. Their sera were assayed for antichlamydial antibodies, and semen analysis and culture were done for each participant. Two hundred and eighty-two (282) male partners of infertile couples were studied. Infertility was commoner among participants aged 40 years or more (45.1%) and was mainly of the "primary type" (62.1%). Antichlamydia antibody was detected in 156 (55.3%) participants and was significantly associated with sperm quality (P = 002; OR = 2.294; 95% CI = 1.36-3.88). Overall, 81 (28.7%) had abnormal sperm quality. The sperm count, progressive motility, and vitality were significantly lower in participants with abnormal sperm quality than those with normal sperm quality (P < 0.001) while morphology, volume, and liquefaction time did not differ significantly (P > 0.05). Staphylococcus aureus was the predominant organism isolated from culture (122/282, 43.3%) while Streptococcus species were the least (4/262, 1.4%). There was significantly more Staphylococcus aureus isolated from the semen of participants that were seropositive to antichlamydial antibodies than those that were seronegative (80/156, 51.3% vs. 42/126, 33.3%; OR = 2.105; 95% CI = 1.30-3.42; P = 0.003). The prevalence of antichlamydial antibodies among male partners of infertile couples in Enugu, Nigeria is high and there is a significant association with sperm quality, sperm count, and bacterial isolates in seminal culture. Male partners of infertile couples in Enugu should be screened for antichlamydial antibodies and appropriate treatment offered wherever indicated. There is a need for increased public awareness and advocacy campaigns on the impact of Chlamydia infection on male factor infertility. This primary preventive measure may help in reducing the burden of Chlamydia infection and male factor infertility.

Dietary patterns and fertility status in men: Mediterranean diet does make a difference in ameliorating the rise in male infertility problems due to changing lifestyle

Dietary patterns and fertility status in men: Mediterranean diet does make a difference in ameliorating the rise in male infertility problems due to changing lifestyle

Spermatophore development in drones indicates the metabolite support for sperm storage in honey bees (Apis cerana).

Developing effective long-term sperm storage strategies to maintain activity requires an understanding of the underlying spermatophore developmental phase in drones. Here we compared the developmental processes and metabolites about seminal vesicles of drones from different parentages (0-24d)in honeybee colonies, including mated queens, virgin queens, and worker bees. The results showed a similar developmental trend of seminal vesicles in thethree groups of drones on the whole, although there were significant differences in developmental levels, as well as in other indicators. Correlation analysis showed significant positive correlations between seminal vesicle width and sperm viability. The metabolomics of the seminal vesicles in drones from mated queens showed differences of the metabolites in each stage. Particularly, squalene identified among them was validated a protective effect on sperm vitality in vitro experiments. Together the results of these assays support that there were significant differences in the developmental levels of seminal vesicles among the three groups of drones in honeybees, wherein a significant correlation between sperm viability and the developmental levels of seminal vesicles were dissected. The metabolomics analysis and semen storage experiments in vitro display signatures of squalene that may act as an effective protective agent in maintaining sperm viability. Collectively, our findings indicate that spermatophore development in drones provides metabolite support, which contributes to research on the differences of sperm viability among drones in the future.

Modified Palomo Procedure Is an Effective Intervention for Improving Serum Testosterone Levels and Semen Parameters in Patients With Varicocele: A Prospective Study.

A varicocele can be defined as an abnormal venous dilatation and/or tortuosity of the pampiniform plexus. It is generally reported that varicoceles are present in 15% of the general male population and 35% of men as a cause of primary infertilityand in up to 80% of men as a cause of secondary infertility. Differences in venous drainage anatomy between the left and right internal spermatic veins, venous valve incompetence resulting in venous blood reflux, and increased hydrostatic pressure are the most commonly cited theories. Various surgical and non-surgical techniques are in use for treating patients with varicocele. Here we used a modified Palamoprocedure to treat the patients and observed the outcome. A total of 40 patients with varicocele were recruited for the study. A preoperative evaluation, along with serum testosterone levels and semen analysis, was done. A modified Palomotechnique was used to treat varicocele. A postoperative follow-up with serum testosterone levels and semen analysiswas done to observe improvement. The mean (±SD) left testis size, right testis size, testosterone, sperm concentration, sperm vitality and sperm progressive motility were found statistically significantly higher in patients after surgery as compared to patients before surgery (p<0.05). However, there was statistically insignificant mean difference in semen volume between patients before surgery and patients after surgery (p>0.05). Modified Palomoprocedure can be used to treat varicocele with good improvement in serum testosterone levels and semen parameters.

Spermiotoxicity of Nano-TiO2 Compounds in the Sea Urchin Paracentrotus lividus (Lamarck, 1816): Considerations on Water Remediation

Despite the great utility of nanoparticles (NPs) in water remediation, their effects on marine ecosystems are unknown and unpredictable. The toxicity of the most used nanoparticles, such as ZnO, Ag, and TiO2 on the purple sea urchin, Paracentrotus lividus (Lamarck, 1816), has been demonstrated by several authors. The aim of this study was to evaluate the effects of TiO2 sol-gel and TiO2-rGO nanocompounds on both vitality and motility of spermatozoa of P. lividus. The spermatozoa were exposed at different times (30 and 60 min) and concentrations (10, 20, 40 µg/mL) of both nano-TiO2 compounds. The results clearly showed a decrease in both vitality and motility of P. lividus spermatozoa exposed. In particular, vitality and motility were inversely related to both exposure time and concentration of TiO2 sol-gel and TiO2-rGO nanocompounds.

Epicatechin Prevents Cryocapacitation of Bovine Spermatozoa through Antioxidant Activity and Stabilization of Transmembrane Ion Channels.

Epicatechin (EPC) is a flavonoid belonging to the family of catechins; it has been described as a powerful scavenger of a wide spectrum of reactive oxygen species (ROS) and a modulator of ex vivo sperm vitality. In this study, we assessed the potential protective abilities of EPC on cryopreserved bovine spermatozoa. We focused on conventional quality parameters, as well as the oxidative profile of spermatozoa alongside capacitation patterns, and expression profiles of proteins involved in the process of capacitation. Semen samples were cryopreserved in the presence of 25, 50 or 100 μmol/L EPC and compared to native semen (negative control) as well as ejaculates frozen in the absence of EPC (positive control). A dose-dependent improvement of conventional sperm quality parameters was observed following EPC administration, particularly in case of the sperm motility, membrane, acrosome and DNA integrity in comparison to the positive control. Experimental groups exposed to all EPC doses presented with a significantly lower proportion of capacitated spermatozoa as opposed to the positive control. While no significant effects of EPC were observed in cases of superoxide production, a significant decrease in the levels of hydrogen peroxide and hydroxyl radical were recorded particularly in the experimental groups supplemented with 50 and 100 μmol/L EPC. Western blot analysis revealed that supplementation of particularly 100 μmol/L EPC to the semen extender prevented the loss of the cation channel of sperm (CatSper) isoforms 1 and 2, sodium bicarbonate cotransporter (NBC) and protein kinase A (PKA), which play important roles in the process of sperm capacitation. In summary, we may hypothesize that EPC is particularly effective in the stabilization of the sperm membrane during the freeze-thaw process through its ability to quench ROS involved in damage to the membrane lipids and to prevent the loss of membrane channels crucial to initiate the process of sperm capacitation. These attributes of EPC provide an additional layer of protection to spermatozoa exposed to low temperatures, which may be translated into a higher post-thaw structural integrity and functional activity of male gametes.

Qiangjing tablets ameliorate asthenozoospermia via mitochondrial ubiquitination and mitophagy mediated by LKB1/AMPK/ULK1 signaling

Context Therapeutic effects of Qiangjing tablets (QJT) on sperm vitality and asthenozoospermia (AZS) have been confirmed. However, the mechanism of action remains unclear. Objective This study investigates the effects of QJT on AZS and the underlying mechanism of action. Materials and methods Sixty Sprague–Dawley rats were randomly divided into six groups: Control, ORN (ornidazole; 200 mg/kg), ORN + QJT-low (0.17 g/mL), ORN + QJT-middle (0.33 g/mL), ORN + QJT-high (0.67 g/mL), and ORN + QJT + Radicicol (0.67 g/mL QJT and 20 mg/kg radicicol) groups. Pathological evaluation and analysis of mitophagy were conducted by H&E staining and transmission electron microscopy, respectively. Reactive oxygen species were detected by flow cytometry. Protein expression was determined by Western blotting. Results QJT significantly improved ORN-treated sperm motility and kinematic parameters, as well as the pathological symptoms of testicular and epididymal tissues. In particular, QJT mitigated impaired mitochondrial morphology, and increased the PHB, Beclin-1, LC3-II protein, and ROS levels (p < 0.05), and reduced the protein expression levels of LC3-I and p62 (p < 0.05). Mechanistically, QJT antagonized the downregulation of SCF and Parkin protein levels (p < 0.05). Furthermore, QJT significantly increased the protein expressions levels of LKB1, AMPKα, p-AMPKα, ULK1 and p-ULK1 (p < 0.05). The ameliorative effect of QJT on pathological manifestations, mitochondrial morphology, and the expressions of mitophagy and mitochondrial ubiquitination-related proteins was counteracted by radicicol. Discussion and conclusions QJT improved AZS via mitochondrial ubiquitination and mitophagy mediated by the LKB1/AMPK/ULK1 signaling pathway. Our study provides a theoretical basis for the treatment of AZS and male infertility.

What Does Intracytoplasmic Sperm Injection Change in Embryonic Development? The Spermatozoon Contribution.

The intracytoplasmic sperm injection (ICSI) technique was invented to solve severe male infertility due to altered sperm parameters. Nowadays, it is applied worldwide for the treatment of couple infertility. ICSI is performed with any available spermatozoon from surgery or ejaculated samples, whatever are the sperm motility, morphology or quantity. The aim of the present review was to study if embryo development and kinetics would be modified by (1) ICSI under the technical aspects, (2) the micro-injected spermatozoa in connection with male infertility. From published data, it can be seen that ICSI anticipates the zygote kinetics Furthermore, because fertilization rate is higher in ICSI compared to conventional in vitro fertilization (IVF), more blastocysts are obtained for clinical use in ICSI. Sperm and spermatozoa characteristics, such as sperm parameters, morphology and vitality, DNA content (levels of sperm DNA fragmentation, microdeletions, and chromosomal abnormalities), RNA content, epigenetics, and sperm recovery site (testicular, epididymis, and ejaculated), have an impact on fertilization and blastocyst rates and embryo kinetics in different ways. Even though ICSI is the most common solution to solve couples' infertility, the causes of male infertility are crucial in building a competent spermatozoa that will contribute to normal embryonic development and healthy offspring.

Donkey semen cryopreservation: Alternatives with permeable, non-permeable cryoprotectants and seminal plasma.

Cryopreservation of semen is an important technique to preserve genetic material. Yet, pregnancy rates in jennies after artificial insemination with frozen-thawed donkey semen are poor. This condition has been attributed to the impact of permeable cryoprotectants, that could cause high post-breeding endometritis. Removal of seminal plasma (SP) prior to semen freezing process is another contributing factor. SP is involved in a multitude of sperm functions and events preceding fertilization and has a mediating effect of sperm capacitation and postcoital uterine inflammatory response. The aim of this study was to evaluate different alternatives in donkey semen cryopreservation with permeable, non-permeable cryoprotectants, BSA and SP. Thirty ejaculates from 10 donkeys were cryopreserved with different combinations of dimethylformamide (DMF, 5%), sucrose (SUC, 200 mM) and homologous SP (10%): DMF (T1), DMF/SP (T2), SUC/BSA (T3), SUC/BSA/SP (T4), DMF/SUC/BSA (T5), DMF/SUC/BSA/SP (T6), DMF/BSA (T7) and DMF/BSA/SP (T8). After thawing, sperm motility and kinetics were assessed by computerized semen analysis. Sperm vitality (SV) was evaluated by fluorescence microscopy, functional membrane integrity (FMI) by the HOST test, abnormal morphology by eosin-nigrosin staining and sperm membrane stability by flow cytometry. For statistical analysis, sperm quality indexes (SQi) were obtained, general linear models were carried out and mean comparisons were made by the Tukey test. T1, T2, T5, T6, and T7 had higher and equivalent results for motility, most kinetic parameters and function membrane integrity. Cryopreservation of donkey semen without permeable cryoprotectant (T3 and T4) showed a reduction in motility, kinetics, SV, FMI and SQi. T5 showed a reduction in progressive motility, sperm velocities, IMF and SQi compared to other DMF treatments. T6 and T8 achieved higher SQi values compared to T1, but they were not different compared to T2 and T7. T1 had a smaller sperm population with low-M540 compared to T3. It is concluded that the use of permeable cryoprotectant is essential to achieve higher post-thaw quality of donkey semen. In addition, the combined use of BSA, SUC and/or PS may provide additional sperm protection compared to the individual use of DMF.