Search Results (93)

Seminal plasma regulates sperm function; oxidative stress (OS) and inflammation can impair semen biochemical homeostasis. This study investigated seminal biochemical/enzymatic compounds and Trolox Equivalent Antioxidant Capacity (TEAC) in seminal plasma and spermatozoa from normozoospermic men (>5th percentile) assessing their relationship with sperm quality. The strongest significant (p < 0.001 ***) negative correlations (r) were found for normal sperm morphology and albumin (ALB, r = −0.515 ***), cholesterol (CHO, r = −0.386 ***), lactate dehydrogenase (LDH, r = −0.514 ***), sperm TEAC (r = −0.438 ***) and homocysteine (Hcy; r = −0.381 ***); sperm progressive motility with ALB (r = −0.495 ***), CHO (r = −0.349 ***), LDH (r = −0.355 ***) and sperm TEAC (r = −0.295 ***); sperm vitality with creatine kinase (CK, r = −0.486 ***) and LDH (r = −0.380 ***). Total prostate-specific antigen (TPSA) was positively correlated with sperm quality. After stratification, subjects with low-normal sperm parameters showed significantly higher levels of ALB, CHO, CK, LDH, interleukin 6 (IL-6), Hcy and sperm TEAC vs. high-normal parameter subjects. ALB, Hcy and LDH were increased in individuals with progressive motility, normal morphology and vitality < 25th percentile indicating their involvement in OS, inflammation and accessory gland activity. These findings suggest that World Health Organization lower reference limits are not a strict cutoff between normal and abnormal semen quality but rather define a baseline of biochemical efficiency and sperm function, highlighting the synergy between spermatozoa and seminal plasma. Full article

Non-obstructive azoospermia (NOA) is characterized by focal and quantitatively limited spermatogenesis, making preoperative prediction of sperm retrieval difficult. Seminal plasma is a biologically plausible liquid-biopsy compartment because it contains testicular, epididymal and accessory-gland secretions enriched with extracellular vesicles, cell-free nucleic acids, proteins and metabolites. This narrative molecular review examines the mechanisms by which germ-cell-derived molecular cargo reaches the ejaculate and organizes seminal-plasma biomarkers by cargo class and spermatogenic stage. Particular attention is given to extracellular-vesicle non-coding RNAs, cell-free seminal mRNAs, germ-cell-enriched proteins including TEX101 and ECM1, and metabolomic and lipidomic signatures. Although several markers show promising discrimination, most remain discovery-stage, single-center and insufficiently validated. The central argument is that the field should move from isolated biomarker nomination toward locked, stage-mapped multi-analyte panels integrated with clinical and genetic predictors under modern prediction-model standards. Seminal plasma is best viewed not as a ready clinical test, but as a biologically coherent platform for future calibrated, externally validated and artificial-intelligence (AI)-ready sperm-retrieval decision support. Full article

The predatory bug Arma custos (Hemiptera: Pentatomidae) is a natural enemy insect capable of preying on over 40 types of agricultural and forestry pests. Here, we describe the characteristics of the morphology and ultrastructure of its venom apparatus visualized using light and electron microscopy. Light microscopy revealed that the venom apparatus of A. custos consists of a pair of main gland and tubular accessory gland. The main gland consist of two lobes, the anterior main gland (AMG) and posterior main gland (PMG). Between the two lobes of the main gland, there is a strong constriction, characterizing a hilum (Hi) where two separate ducts, the venom duct of the main gland (VD) and the duct connecting the accessory gland to the main gland (AMD), are inserted. The VD extends toward the head and connects to the venom pump (VP), while the AMD extends toward the thorax and connects to the accessory gland (AG). Ultrastructural examination of the venom glands reveals that the AMG and PMG consist of a layer of cubic or spherical glandular cells forming a large circular lumen, while the AG exhibits two narrow lumens. The secretory cytoplasm of AMG, PMG, and AG contains a well-developed rough endoplasmic reticulum, along with mitochondria, nuclei, secretory vesicles, autophagosomes, and secretory granules. However, significant differences exist in the ultrastructural characteristics among the three glands. Unlike glandular secretory cells in the venom glands, the ultrastructure of VD, and AMD reveals only well-developed nuclei, mitochondria, and elaborate plasma membrane folds. These results indicate that venom proteins are synthesized and stored by the AMG, PMG, and AG, while the VD and AMD ducts are responsible for transporting the venom. Full article

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. Full article

Typically, most omics analysis (proteomic and transcriptomic) of snakes are focused on the dominant enzymatic proteins used for evolutionary analysis or those engaged in envenoming symptoms. This study presents a comprehensive multi-assembler transcriptomic analysis focused on the non-dominant and enzymatic or non-enzymatic putative proteins of the venom glands of three medically significant Colombian snake species. Together, these results highlight how continued improvements in modern omics workflows, coupled with extensive manual curation, enable more complete putative protein variants discovery when multiple assemblers are integrated. Here, we reconstructed the toxinomes of the viperids Bothrops asper and Crotalus durissus cumanensis, and the elapid Micrurus mipartitus, by comparing four assemblers (Trinity, SPAdes, SOAPdenovo-Trans k = 31 and k = 97) and integrating them into a non-redundant meta-assembly. Protein-candidate alignments were extensively inspected, and validation of conserved domains and functional motifs are discussed. The curated toxinomes revealed substantial diversity across major and accessory families, and assembler choice strongly affected transcript variant recovery. Together, these results provide a more comprehensive view of venom-gland transcriptome analysis and diversity, expanding the set of candidate venom components for future functional and proteomic validation, with potential implications for venom composition studies and antivenom development. Full article

The vomeronasal system (VNS) is directly linked to the various behavior and ecology of all animal species, and understanding it might help to prevent deer damage. We therefore histochemically analyzed the accessory olfactory bulb (AOB) and the vomeronasal organ (VNO) that, respectively, function as a primary integrative center and a peripheral receptor organ, in Hokkaido sika deer (Cervus nippon ssp. yesoensis). The AOB consisted of the vomeronasal nerve, glomerular, plexiform, and granule cell layers. We found that G protein α subunit i2 (Gαi2) and o (Gαo) that are, respectively, coupled with vomeronasal receptor type 1 and 2 families were strongly and weakly expressed in the glomerular layer, respectively. These properties of the AOB of sika deer were similar to those of other artiodactyl species, including wapiti. We then explored the sika deer VNO using 21 lectins that bind to each glycan structure. Although various cell types in the VNO had unique lectin binding profiles, all 21 lectins bound to the free border of the sensory epithelium, suggesting that various glycoconjugates are involved in pheromone detection in sika deer via the VNO. Furthermore, the reactivity of some lectins in the sensory epithelium and vomeronasal gland differed from those of roe deer and wapiti. Our findings suggest that the composition of glycoconjugates in the VNO differs among deer species. Full article

Background: Congenital cheek sinuses are exceedingly rare craniofacial anomalies, typically present at birth and often misdiagnosed due to their resemblance to dermal pits or dimples. Only a few cases have been reported in the literature. This case is notable for its histopathological confirmation as a type II first branchial cleft anomaly containing an accessory tragus, highlighting the embryologic and clinical overlap between congenital cheek sinuses and branchial cleft anomalies. Case presentation: We report a three-year-old boy presenting with a congenital dimple along the right nasolabial fold. The lesion was present since birth and occasionally discharged clear fluid but had no history of infection or inflammation. Imaging demonstrated a subcutaneous tract without communication to the oral cavity or parotid gland. Under general anesthesia, surgical excision was performed, and the sinus tract was dissected in continuity to a cartilaginous remnant. Histopathologic analysis confirmed the presence of an accessory tragus consistent with a type II first branchial cleft anomaly. The patient’s postoperative recovery was uneventful, and at three months’ follow-up, there was no recurrence and an excellent cosmetic outcome. Conclusions: This case emphasizes the importance of considering branchial cleft anomalies in the differential diagnosis of congenital cheek sinuses. Although these lesions may appear benign and superficial, histopathological confirmation is critical for accurate classification. Complete surgical excision remains the definitive treatment, ensuring both diagnostic clarity and favorable aesthetic results. Increased awareness of such rare anomalies can improve diagnostic accuracy and surgical planning, particularly with respect to their potential proximity to critical facial structures. Full article

Alpha-lipoic acid (ALA) is well-known for its antioxidant and anti-inflammatory functions in various disease models. Here, we tested whether pre-exposure to ALA can protect the testes from cellular damage caused by scrotal heat shock (HS) in mice. Methods: Thirty-six Swiss albino mice were divided into control (CTRL, n = 6), HS (n = 10), and two ALA dose (HS + ALA 200 mg/kg, n = 10; and HS + ALA 400 mg/kg, n = 10) groups. ALA supplementation was administered orally for 30 days. Subsequently, the animals, except the controls, were subjected to an HS water bath at 43 °C for 20 min. Two days later, they were euthanized, and biometric data from gonads and accessory sexual glands, testicular samples for histomorphometric and immunohistochemical analyses, and sperm from the epididymis cauda were obtained for evaluation. Results: Animals submitted to HS had a lower body weight, decreased relative mass of testes and prostate, reduced seminiferous epithelium height and tubular diameter, and increased degeneration in seminiferous tubules. Additionally, sperm analysis showed a reduced linear progressive velocity (VSL) and straightness (STR), increased midpiece defects, and fewer sperm with functional membranes. Immunohistochemical evaluation revealed a reduced superoxide dismutase (SOD1) staining intensity in the testes. Preventive exposure to ALA at 200 mg/kg did not normalize the relative testicular mass, but it reduced the number of giant cells, decreased midpiece defects, normalized the number of sperm with functional membranes, and partially preserved SOD1 expression. Although animals treated with ALA 400 mg/kg showed an improvement in relative testicular mass, this dose was less efficient in other parameters. Conclusions: This study showed that while 30 days of oral ingestion of ALA before the induction of acute degenerative injury did not fully protect male mouse gonads at the tissue level, some parameters related to testicular function and sperm quality showed a partial improvement. Full article

Cathepsins represent a crucial group of protein enzymes involved in insect metabolism. Within the Hemiptera order, comprising a diverse array of predatory, blood-feeding, and herbivorous species, the understanding of cathepsin types and their roles as venom components in predatory bugs remains limited. This investigation systematically identified cathepsin genes present in Hemiptera genomes, highlighting a prevalence of cathepsin B and L, with cathepsin D exhibiting a higher gene count in the Heteroptera suborder. Examining the predatory assassin bug Sycanus bifidus, eight cathepsin genes were notably expressed in its venom glands, with the SbCAB2 gene from the cathepsin B subfamily demonstrating the highest expression in the posterior main gland, indicating its significance as a venom component. Subsequent expression and purification of the recombinant SbCAB2 protein revealed heightened hydrolytic activity (0.91 U/mg protein) compared to extracts from the anterior main gland, accessory gland, and gut. Functional assays demonstrated that SbCAB2, at lower doses (0.625–2.5 μg), can impede phenoloxidase activity in Tenebrio molitor pupal hemolymph, with a 2.5 μg dose inhibiting 86.5% of this activity, thereby preventing hemolymph melanization. Conversely, a higher dose of 10 μg led to effects akin to human placental cathepsin B, promoting melanization in T. molitor pupal hemolymph. These findings lay the foundation for further exploration of the adaptive evolution of cathepsin genes in Hemiptera and offer crucial insights into the functional role of venomous cathepsins in predatory bugs. Full article

In most mosquito species, reproduction requires mating between the female and the male, followed by the female blood-feeding, completing oogenesis, and laying eggs. Warmer environmental temperature and aging both reduce mosquito fecundity and fertility, and warmer temperature accelerates the aging-dependent decline in reproduction such that reproductive impairment manifests earlier in life. To shed light on how this warming-based acceleration of reproductive senescence occurs, we investigated how temperature (27 °C, 30 °C, and 32 °C) and aging interactively shape female and male reproductive tissue size in the African malaria mosquito, Anopheles gambiae. In blood-fed females, we discovered that warmer temperature accelerates the aging-dependent decrease in the size of the ovaries but not the spermatheca. In males, we discovered that warmer temperature lessens and delays the aging-dependent increase in the size of the male accessory glands but not the testes. Next, we measured the expression of reproductive genes in females and males. In female reproductive tissues, warmer temperature accelerates the aging-dependent decrease in the expression of vitellogenin and the aging-dependent increase in the expression of MISO and HPX15. In male reproductive tissues, warmer temperature accelerates an aging-dependent decrease in the expression of Plugin, TGase3, phLP, and CYP315A1. Altogether, these data shed light on how physical and transcriptional changes underpin the warming-based acceleration of an aging-dependent decline in mosquito fecundity and fertility. Full article

The prostate gland, a male accessory reproductive organ, is regulated by hormonal inputs and autonomic innervation from the major pelvic ganglion. This study examined the effects of major pelvic ganglion denervation on prostate histology, immune cell infiltration, and systemic levels of prolactin, testosterone, and cytokines in rats. Male Wistar rats (300–350 g) were divided into groups receiving bilateral axotomy of the hypogastric nerve, the pelvic nerve, or both, alongside with a sham-operated control. After 15 days, the animals were killed, and prostate tissue was dissociated in DMEM medium containing DNase I and collagenase. The dissociated cells were stained with fluorochrome-conjugated antibodies, and cell characterization was performed using a flow cytometer. Hematoxylin and eosin (H&E) staining was used to analyze histological characteristics, while testosterone, prolactin, and interleukin levels were measured via ELISA. Histological analysis revealed inflammatory atypical hypertrophy e hiperplasia. Immunological assessments demonstrated increased leukocytes, T lymphocytes (CD4⁺ and CD8⁺), B lymphocytes, and macrophages following double nerve axotomy. Serum analyses showed elevated pro-inflammatory cytokines IL-1β, IL-6, and IFN-γ, as well as anti-inflammatory IL-10, in denervated animals. Hormonal assessments revealed significant increases in serum prolactin and testosterone levels after double axotomy. Loss of neural control may promote pathological prostate changes via inflammation and hormonal dysregulation, offering insights into neuroimmune and neuroendocrine mechanisms underlying prostate pathologies. Full article

Grapholita molesta is a globally significant fruit pest. Females achieve maximal reproductive output through efficient sperm utilization following a single copulation. Post-mating maturation of eupyrene sperm is a critical step in reproductive success. Here, we report that a male accessory gland-derived serine protease (named GmAGSP1) is essential for this process. GmAGSP1 was only distantly related to other identified sperm-activating SPs, and its transcript was highly expressed in the AG at 48 h after emergence. RNAi-mediated knockdown of GmAGSP1 in males did not affect courtship rate, copulation duration, or mating frequency, whereas male fertility decreased significantly. Mating with GmAGSP1-knockdown males markedly impaired eupyrene sperm maturation in the spermatophores, with phenotypes including failure of eupyrene sperm bundles to dissociate normally and marked reduction in viability of the dissociated eupyrene sperm. Finally, untargeted metabolomic analysis preliminarily demonstrated marked alterations in multiple metabolic pathways within the spermatophore following mating with GmAGSP1-knockdown males. This study advances our understanding of the regulatory mechanism of “sperm activation in the spermatophore’s metabolic microenvironment mediated by male AG-derived SP” while providing critical insights for the development of novel genetic control strategies targeting G. molesta. Full article

(1) Background: The insect male accessory gland (MAG) produces seminal fluid components crucial for male reproduction, analogous to the mammalian prostate. While some insect MAGs exhibit binucleate epithelial cells for luminal volume plasticity, the diversity of cellular arrangements and their functional implications across insects remain largely unknown. (2) Methods: We investigated the cellular architecture of MAG epithelia in various shore bug species (infraorder Leptopodomorpha, Hemiptera) and their mechanisms of multinucleation and potential MAG volume regulation. (3) Results: The MAG epithelia of shore bugs comprise a small number of large, plastic syncytial cells with varying nuclear numbers. We hypothesize that these syncytia facilitate effective MAG volume expansion post-eclosion. Uniquely, MAG shrinkage involves the localized contraction of limited muscle fibers, unlike the systematic contraction of circular muscles in most other insects. We further describe sequential cell fusion during the nymphal stage as the mechanism of multinucleation. (4) Conclusions: The unique syncytial organization of Leptopodomorpha MAG epithelia represents an evolutionary divergence from typical binucleate or mononucleate structures in other insects; it is likely that this enables distinct mechanisms for reproductive fluid storage and evacuation. This study highlights the evolutionary diversity of male reproductive organ morphology and function within insects. Full article

Male tract infections (MTIs) are a common clinical condition, often presenting without any signs nor symptoms of disease. As advised by the European Urology Guidelines dealing with this topic, patients are typically treated with antibiotics alone. Nevertheless, in between 40% and 50% of cases, antibiotic therapy is not effective in eradicating the semen infection. Therefore, persistent semen infection is frequently found upon semen culture evaluation following antibiotic therapy. In this study, we aimed to analyze the fecal microbiota of male infertile patients with persistent MTI in order to verify the prevalence of gut dysbiosis in these patients. We therefore enrolled 20 infertile patients with persistent MTIs after a proper cycle of antibiotic treatment. All patients performed the study for gut microbiota analysis after about 30 days after the last dose of antibiotic treatment. Gut microbiota analysis revealed that 50% of patients with persistent MTI presented a reduction in microbial biodiversity. Indeed, a situation of gut dysbiosis was reported in 75% of patients. In details, the Firmicutes–Bacteroidetes ratio was reduced in 70% of such patients, including 40% of patients where a severe reduction was observed due to an elevated abundance of Bacteroidetes (putrefactive dysbiosis). The most frequent enterotype was Prevotella-dominant (43%). We demonstrated for the first time that patients with recurrent MTIs have enterotypes associated with increased gut permeability and systemic inflammation. Further studies are required to analyze the molecular machinery by which gut dysbiosis exerts its role in patients with MTIs, in particular persistent MTIs, and how supplementation with probiotics might impact in terms of restoring eubiosis, in terms of eradicating the infection, and reducing prostate inflammation and eventually in terms of improving semen evaluation in male infertile patients. Full article

The brown planthopper Nilaparvata lugens, a major rice pest, threatens global food security through rapid reproduction. This study investigates the role of the tramtrack (ttk) gene in male reproductive development and spermatogenesis using RNA interference (RNAi). Gene expression analysis revealed higher ttk levels in testes. RNAi-mediated knockdown of ttk in fourth-instar male nymphs reduced its expression by up to 80%, leading to severely impaired gonad development. Testes, vas deferens, and accessory glands in treated males exhibited 8–89% volume reductions compared to controls, accompanied by a 51–69% decline in sperm count and 60–85% reduction in sperm motility. Consequently, eggs fertilized by treated males showed a 73% decrease in hatching rates, with arrested embryonic development. These findings demonstrate ttk’s critical role in spermatogenesis and gonad maturation in N. lugens, highlighting its potential as an RNAi target for sustainable pest control strategies. Full article