Phosphatidylcholine promotes sperm motility by regulating sperm membrane phospholipid homeostasis.

Blood-testis barrier-crossing extracellular vesicles for asthenozoospermia therapy via synergistic ATP replenishment and ferroptosis suppression.

To assess the potential utility of leukocytes and spermatozoa telomere length (LTL and STL) as reproductive biomarkers, this study measured both LTL and STL and investigated their possible correlations with oxidative status and semen quality parameters in healthy breeding male dogs. Twenty-two ejaculates and blood samples were collected from dogs of various breeds. Semen was evaluated for volume, concentration, sperm motility, and kinetic parameters using a Sperm Class Analyzer. LTL and STL were quantified using Quantitative PCR (qPCR) and expressed as the relative ratio of telomere repeat copy number (T) to a single-copy gene (S), T/S ratio. Serum oxidative stress markers were assessed using d-ROMs and Biological Antioxidant Potential (BAP) tests. Correlations between STL and LTL, as well as their association with age, semen parameters and oxidative stress levels, were evaluated using Spearman analysis. LTL and STL were 0.63±0.25 and 0.85±0.31 and showed a strong positive correlation (P<0.001; rs=0.70). Both LTL and STL were also negatively correlated with age (P<0.05 and rs=-0.50; P<0.001 and rs=-0.68). The cut-off age for the difference in STL and LTL was identified at 6 and 7.5 years, respectively. STL was also positively correlated with semen volume (P<0.05; rs=0.63) and concentration (P<0.05; rs=0.41) and negatively correlated with semen chromatin decondensation (P<0.01; rs=-0.71); while the LTL showed a positive correlation with sperm concentration (P<0.05; rs=0.53). No correlations with oxidative markers were found. These findings support the potential use of TL as a biomarker for reproductive aging and semen quality in dogs.

Effects of intraperitoneally administered gadolinium on male reproductive tissues and functions in wistar rat.

Advances in sperm transcription and translation activities: From traditional views to new discoveries.

Co-exposure to polystyrene nanoplastics and hexachlorocyclohexane induces enhanced human sperm toxicity in vitro.

Male immunological infertility (MII), driven by anti-sperm antibody (AsAb), is a complex autoimmune disorder. The herbal formulation TuominJian (TMJ) has shown clinical efficacy against MII, but its immunomodulatory mechanisms involving Tfh cells, B cells, and cytokines remain unclear. This study aimed to elucidate the immunoregulatory effects and underlying mechanism of TMJ in MII mice. Active compounds in TMJ were identified using UPLC-Q/TOF-MS. An MII mouse model was established using sperm immunization. After verifying the model, testicular and epididymal tissue integrity, sperm parameters, and blood-testis barrier function were assessed via H&E staining, TEM, CASA, and western blotting. Serum AsAb, cytokines, and immune cell populations were analyzed by ELISA and flow cytometry. Thirteen active compounds were identified in TMJ. TMJ treatment significantly reduced testicular cell apoptosis and decreased serum AsAb and proinflammatory cytokines (IL-1, IL-6). It also restored testicular and epididymal tissue architecture, improved sperm concentration and motility, and enhanced blood-testis barrier integrity by upregulating occludin. Furthermore, TMJ downregulated Tfh and B cell levels. TMJ mitigates MII in mice by promoting structural recovery of reproductive tissues, reinforcing the blood-testis barrier, and modulating immune responses through suppression of Tfh cells, B cells, and proinflammatory cytokines. These findings highlight immunoregulation as a promising therapeutic strategy for MII.

This study investigates the effects of graphene oxide (GO) on the quality of frozen-thawed ram semen. Each of 30 ejaculates from five rams were cryopreserved with a control extender (Optidux®, Reprodux Inc., Brazil) or extender containing GO concentrations at 5, 50, or 100 µg/mL. Post-thaw data were analyzed using Tukey's or Dunn's t test (p < 0.05). Graphene oxide at 5 and 50 µg/mL increased frozen-thawed total (50.4 ± 3.15; 52.0 ± 2.76 %), and progressive sperm motility (44.2 ± 3.06; 45.7 ± 2.68 %) vs control (42.0 ± 2.75; 36.7 ± 2.58 %), and sperm viability (56.3 ± 2.17; 57.8 ± 2.82 %) vs control (48.8 ± 2.60 %); and decreased immotile spermatozoa (49.6 ± 3.15; 48.0 ± 2.76 %) vs control (58.0 ± 2.75 %). Both 5 and 100 µg/mL treatments improved DNA integrity (90 ± 0.34; 90.5 ± 0.44 %) vs control (89.1 ± 0.37 %). There were no GO effects on sperm morphology, VCL, VAP. Local motility, an undesirable parameter, increased significantly from 5.42 ± 0.52 % (control) to 6.77 ± 0.43 % (100 µg/mL GO). Supplementation with GO improved post-thaw sperm progressive motility (by up to 24.5 %) and viability (up to 18.4 %), reducing immotile sperm (up to 17.2 %), compared to extender alone. This potentially supports the practical use of GO as an additive to semen extenders.

The capercaillie (Tetrao urogallus), a critically endangered species, faces significant challenges due to habitat loss and population decline. Assisted reproductive technologies, including semen preservation and artificial insemination (AI), are vital for conservation efforts. Although the results are controversial, it has been reported that antioxidants, such as catalase, play an important role in protecting against oxidative damage caused by reactive oxygen species during short- and long-term sperm storage. This study evaluated the protective effects of catalase (200 IU/mL) on stored/frozen-thawed capercaillie sperm. The possible protective or harmful effect of catalase on fertilizing ability was assessed after AI with fresh and stored semen. Results indicated that catalase had no significant effect on sperm motility, viability, or DNA integrity after liquid storage or freezing/thawing. Fertility rates decreased sharply after semen storage for 6 h at 5°C even in the presence of catalase. Catalase-treated samples maintained similar fertility rates to controls. The study also provided the first evidence of sperm storage duration within female capercaillies, which lasted up to 21 days. These findings underscore the challenges of semen preservation in wild species and highlight the need for further exploration of alternative antioxidants to optimize storage media.

Paclitaxel (PTX) is broadly prescribed to treat various malignancies. However, it induces negative impacts on many organs, including testes. This study explored the beneficial role of sitagliptin (SIT) in PTX-provoked testicular damage and the underlying mechanisms. Rats were allocated into four groups: (I) control, (II) PTX, (III) PTX + SIT5, and (IV) PTX + SIT10. Histopathological and ultrastructural analyses were conducted along with sperm analysis. Immunohistochemical examinations of NOD-like receptor protein 3 (NLRP3), cleaved caspase-3, caspase-3, cytochrome c (Cyt.c), and interleukin-1 beta (IL-1β) were assessed. Serum testosterone and testicular 17β-hydroxy steroid dehydrogenase (17β-HSD), sestrin2, phosphorylated protein kinase R-like ER kinase (pPERK), and C/EBP homologous protein (CHOP) were determined. SIT induced a remarkable increase in sperm count, motility, and viability, with a pronounced decline in sperm abnormality compared to PTX group. SIT increased testosterone and 17β HSD levels. SIT elevates sestrin2, reduced glutathione (GSH), and catalase, and reduces malondialdehyde (MDA), reflecting its antioxidant action. SIT mitigates ER stress via diminishing pPERK and CHOP. SIT reduces NLRP3 and IL-1β levels, clarifying its anti-inflammatory action. SIT decreases cleaved caspase-3, caspase-3, and Cyt.c levels, verifying its anti-apoptotic features. Overall, SIT ameliorated PTX-provoked testicular dysfunction via mediating PERK/CHOP/NLRP3/Sestrin2 signaling pathway.

Mass motility assessment is widely used to evaluate semen quality in the sheep breeding industry, yet the biological factors influencing motility duration remain poorly characterised. At ejaculation, spermatozoa are combined with seminal plasma (SP), which initially supports motility and function but may compromise long-term metabolic endurance. This study aimed to investigate the effects of SP on mass motility duration and pH regulation of epididymal ram spermatozoa. Epididymal sperm were incubated with whole SP, saline, or left undiluted to assess motility duration. Subsequently, sperm were incubated with varying SP concentrations and molecular weight-separated fractions (protein-free <3kDa; protein-enriched >3kDa) and assessed for motility and pH over 24h. Metabolomic and lipidomic profiling via targeted mass spectrometry characterised the biochemical composition of each SP fraction. SP exposure significantly reduced mass motility duration (195±11min) compared with undiluted sperm (1797±185min; P=0.0123) and saline dilution (2667±134min; P<0.0001). All SP-treated groups exhibited a progressive pH decline, consistent with increased glycolytic flux, whereas SP-naïve sperm showed stable or increasing pH, indicating a more regulated metabolic state and greater metabolic flexibility. The protein-free SP fraction was most detrimental to motility, which was likely to be the result of the depletion of key proteins, lipids, and metabolites associated with energy production. Prolonged exposure to SP impairs ram sperm motility longevity by promoting a glycolysis-biased metabolic state and reducing metabolic endurance. Understanding how seminal plasma composition influences sperm metabolic regulation may inform strategies to better preserve sperm motility during semen handling and storage.

Vasovasostomy (VV) is often pursued by men seeking natural conception after vasectomy. Microsurgical VV is associated with high patency rates (~95%) and moderate pregnancy rates (~40%), according to existing literature. However, long-term outcome data remain limited. This review evaluates patency and pregnancy rates at ≥12 months following microsurgical VV. A comprehensive literature search was conducted via PubMed using the terms "vasovasostomy," "patency rate," and "pregnancy rate." Studies were included if they reported patency or pregnancy outcomes ≥12 months post-VV. Data were categorized and analyzed using MedCalc, applying the Freeman-Tukey transformation for normalization. Only four clinical studies reported long-term patency data, and six studies reported long-term pregnancy rates following microsurgical VV. Patency rates ranged from 77% to 99%, with a mean follow-up of 25 months. Patency was defined as the presence of any sperm in the ejaculate in five studies, and as >1 million non-immotile sperm/mL in one study. Using the >1 million sperm definition, the patency rate was 77% at 12 months, but decreased to 33% when defined as ≥30% motile sperm in the ejaculate. Pregnancy rates across the six studies ranged from 28% to 54%, with a mean follow-up of 21 months. Long-term patency and pregnancy rates following VV vary widely due to inconsistent definitions and limited follow-up data. These findings underscore the need for standardized outcome measures and longitudinal follow-up to better understand factors influencing long-term success after VV.

The aim of this study was to compare outcomes of microsurgical varicocelectomy (MV) versus percutaneous angioembolization (AE) in men with varicoceles causing infertility or scrotal pain. Endpoints included postoperative changes in semen parameters, complication rates, pain relief, need for additional interventions, and the influence of body mass index (BMI). We retrospectively reviewed 264 male patients treated for varicocele from July 2015 to July 2024 (MV: 214 patients, and AE: 50 patients). Only patients older than 18 years were included. Indications were infertility or chronic varicocele-related scrotal pain. Pre- and post-treatment semen analyses were compared in infertile men, and pain outcomes were assessed in those treated for pain. Complications and secondary interventions were recorded, and outcomes were stratified by BMI. MV significantly improved sperm concentration, motility, and total motile count within 3-6 months, while AE showed no significant change. Complications occurred only in the MV group (8.9%), including hydrocele, hematoma, and surgical site infection, while no complications were reported in the AE group. Persistent pain was observed in one-third of patients in both groups, and AE was associated with a higher rate of secondary intervention (14.0%) compared to MV (6.6%). BMI was not linked to complications or operative time but was associated with poorer baseline semen quality. Within 3-6 months post-treatment, varicocelectomy yielded greater improvement in semen quality, while AE offered a favorable safety profile but less reproductive benefit and a higher retreatment rate. Both approaches were safe across BMI categories, though longer-term studies are needed to clarify patient selection and delayed outcomes.

Despite advances in reproductive medicine, the role of microbes in infertility remains underexplored. Our earlier research demonstrated an Escherichia coli strain, capable of causing 100% sperm immobilization via agglutination in-vitro, and infertility in-vivo. Also, sperm agglutination factor (SAF), extracted and purified from bacterial culture, depicted similar results and, showed homology to glutamate decarboxylase. However, it remained unclear whether such bacterial protein signatures are produced within the vaginal milieu post-colonization of E. coli in host environment. Addressing this, discovery-phase proteomics approach was used to investigate vaginal lavage fluid (VLf) of female mice following intravaginal administration of E. coli. VLf from E. coli-administered and PBS-treated groups revealed distinct protein banding patterns (SDS-PAGE) and chromatographic profiles (RP-HPLC). Proteomic-profiling of pooled VLf using nano-LC-MS/MS depicted a ~ 20-kDa bacterial protein, annotated as Uncharacterized protein YfgI, only in test group. Gel-filtration chromatography of VLf revealed a purified protein of ~ 20-kDa that aligned with the proteomic analysis and hence, designated as VLf-protein. Functionally, VLf-protein impaired sperm motility, viability, induced ultrastructural abnormalities, demonstrated binding to sperm surface, and infertility in-vivo. Structural modeling and comparative analysis revealed only suggestive similarity between VLf-protein and SAF.

Male accessory gland inflammation (MAGI) can impair male fertility through inflammation-driven oxidative stress and direct sperm damage; nutraceutical approaches may be useful when antibiotics are not indicated. Here, we evaluated a 3-month treatment with a Graminex™-based dietary supplement (Deprox-HP) in twenty MAGI patients integrating conventional semen analysis and oxidative stress assessment with sperm proteomics before and after therapy. After treatment, total and progressive sperm motility increased significantly, whereas sperm concentration and sperm morphology showed a non-significant upward trend. Sperm lipid peroxidation decreased markedly, while the antioxidant capacity showed a non-significant increase. Analysis of the sperm proteome demonstrated a clear PRE-POST clustering, consistent with treatment-associated remodeling. POST samples showed upregulation of proteins linked to sperm motility, redox homeostasis, mitochondrial metabolism and membrane remodeling. Two pregnancies occurred during the treatment period; in both cases, lipid peroxidation decreased along with an increase of morphologically typical spermatozoa, and sperm proteomics showed a concordant post-treatment shift enriched in flagellar and mitochondrial respiratory/redox compartments. Moreover, we found a selective enrichment POST treatment in these two patients of TEX50, a crucial protein involved in acrosome/head-stability during epididymal transit. Overall, Deprox-HP was associated with reduced oxidative membrane damage and a coordinated sperm proteomic shift consistent with improved motility.

ABSTRACT Silica nanoparticles (SiO 2 NPs) are widely used in biomedical and industrial applications, raising concerns about chronic exposure effects. This study investigated the impact of 90‐day oral SiO 2 NP exposure on male reproductive function in Wistar rats ( n = 6). In accordance with OECD 408 guidelines and our 14‐day range‐finding study, doses of 500, 1000, and 2000 mg/kg were selected, with 2000 mg/kg identified as the highest nontoxic level for sub‐chronic exposure, and reproductive endpoints including hormone levels, sperm quality, and testicular histopathology were subsequently evaluated. Exposure resulted in significant dose‐dependent reductions in body weight, food intake, reproductive organ weights, and serum hormone levels such as testosterone (T), follicle‐stimulating hormone (FSH), and luteinizing hormone (LH) ( p &lt; 0.001), accompanied by histopathological damage to testicular tissue. Sperm count, vitality, motility, and morphology were significantly and dose‐dependently impaired. Enzymatic activities of testicular biomarkers‐including acid phosphatase (ACP), glucose‐6‐phosphate dehydrogenase (G6PD), γ‐glutamyl transferase (γ‐GT), and succinate dehydrogenase (SDH) were significantly decreased (&lt; 0.001), suggesting metabolic dysfunction. Real‐time PCR analyses revealed downregulation of SF‐1 and associated steroidogenic genes (HSD3B1, HSD17B10, StAR), which was associated with altered testosterone biosynthesis. The observed SF‐1 downregulation was associated with altered steroidogenic regulation, possibly mediated through oxidative and endocrine stress pathways. This study highlights the importance of assessing the long‐term safety of SiO 2 NPs and their impact on reproductive health, particularly in the context of SF‐1‐mediated mechanisms.

The objective of the present study was to evaluate the effect of dietary supplementation with the prebiotic Saccharomyces cerevisiae yeast on testicular width and semen quality in rabbit bucks. Twenty-four male rabbits were randomly assigned to three groups and their semen was sampled for 8 weeks: a control group (C) fed with a conventional diet without S. cerevisiae, a group supplemented with 0.3 g of S. cerevisiae/kg of feed (T1) and a group supplemented with 1 g of S. cerevisiae/kg of feed (T2).Testicular width was determined using a caliper. Semen was collected from each buck and evaluated for sperm volume, motility, percentage of live sperm and morphological characteristics. Theresults showed that both right and left testicles were greater (P < 0.01) in width in group T1. Regarding semen parameters, progressive sperm motility was higher (P < 0.01) in the T2 group compared to the C and T1 groups. A significant decrease (P < 0.01) in the percentage of spermatozoa with abnormal mid-piece and flagellum was observed in the T2 group compared to the other groups. Furthermore, the percentage of undeveloped sperm decreased (P < 0.01) in both T1 and T2 groups. However, semen volume and percentage of live sperm did not vary between groups (P > 0.05). Theseresults suggest that S. cerevisiae improved the progressive sperm motility and morphological characteristics at the rate of 1 g kg-1 of feed.

Oligo-astheno-teratozoospermia (OAT), a recurrent cause of male infertility, is the most frequent disorder of spermatogenesis with a predominantly genetic origin. Patients and mice bearing mutations in the ARMC2 gene exhibit reduced sperm concentration, multiple morphological defects, and impaired motility, defining a canonical OAT phenotype. Intracytoplasmic sperm injection (ICSI) is required to treat this condition; however, it is associated with a slightly increased risk of birth defects compared with natural conception, highlighting the need for novel targeted therapies. Here, in vivo testicular injection followed by electroporation of capped, polyadenylated naked messenger RNA (mRNA) was evaluated as a strategy to treat ARMC2 -related infertility in mice. mRNAs encoding reporter proteins were used to assess expression efficiency and kinetics using in vivo and in vitro 2D and 3D imaging. Reporter proteins were detected in germ cells for up to three weeks, demonstrating the feasibility of mRNA-based approaches. These results were compared with a non-integrative plasmid Enhanced Episomal Vector, which induced weak and transient expression in spermatogenic cells. Delivery of Armc2 mRNA restored morphologically normal and motile sperm in deficient males, capable of producing embryos via in vitro fertilization and ICSI. These findings provide proof-of-concept that mRNA electroporation can restore sperm motility and fertilizing potential, offering a novel strategy to correct monogenic male infertility.

Oligo-astheno-teratozoospermia (OAT), a recurrent cause of male infertility, is the most frequent disorder of spermatogenesis with a predominantly genetic origin. Patients and mice bearing mutations in the ARMC2 gene exhibit reduced sperm concentration, multiple morphological defects, and impaired motility, defining a canonical OAT phenotype. Intracytoplasmic sperm injection (ICSI) is required to treat this condition; however, it is associated with a slightly increased risk of birth defects compared with natural conception, highlighting the need for novel targeted therapies. Here, in vivo testicular injection followed by electroporation of capped, polyadenylated naked messenger RNA (mRNA) was evaluated as a strategy to treat ARMC2-related infertility in mice. mRNAs encoding reporter proteins were used to assess expression efficiency and kinetics using in vivo and in vitro 2D and 3D imaging. Reporter proteins were detected in germ cells for up to three weeks, demonstrating the feasibility of mRNA-based approaches. These results were compared with a non-integrative plasmid Enhanced Episomal Vector, which induced weak and transient expression in spermatogenic cells. Delivery of Armc2 mRNA restored morphologically normal and motile sperm in deficient males, capable of producing embryos via in vitro fertilization and ICSI. These findings provide proof-of-concept that mRNA electroporation can restore sperm motility and fertilizing potential, offering a novel strategy to correct monogenic male infertility.

Spermatogenesis is worsened by reactive oxygen species (ROS) and antioxidants could reduce ROS induced sperm damage. This prospective cohort study evaluated the potential therapeutic effects of a combination of vitamins, minerals and antioxidants on the sperm quality parameters of infertile men with idiopathic low spermatogenesis. Seminal fluid analysis tests were performed before treatment, 3 and 6 months after treatment. The treatment resulted in a significant improvement in the rate (%) of sperm motility from 16.95±6.93 to 23.11±8.87, after 3 months and reached 23.68±8.73 after 6 months (p=0.0006) whereas a non-significant increase in sperm count (from 13.05±8.1 to 15.79±7.9 after 3 months and 15.26±10.3 million/ml after 6 months (p=0.1650). Morphology and agglutination showed little changes. A positive correlation between sperm count and motility was observed after 3 months of treatment (r = 0.594; p=0.007). The combination of vitamins and antioxidants improved the sperm motility significantly and to a lesser extent the sperm count, however, sperm morphology and agglutination remained relatively unchanged. Using antioxidants is safe and can improve semen parameters.