This study explores the endocrine-disrupting effects of climbazole (CBZ), an environmental and lifestyle stressor, on male fertility. The impact of CBZ on sperm vitality, motility, and molecular pathways related to hormone receptors and apoptosis was evaluated, in non-capacitated and capacitated conditions. Gene expression of key components, including hormone receptors (ESR1, ESR2, FSHR, AR), apoptosis-related genes (BAX, BCL2), and COX4l1 (involved in mitochondrial function), was analyzed. Protein tyrosine phosphorylation, a marker of capacitation, was also examined using immunofluorescence and Western blot analysis. We demonstrated that CBZ significantly reduced sperm vitality at concentrations above 25 µM and motility at 1 and 10 µM in non-capacitated and capacitated conditions. Changes in tyrosine phosphorylation patterns were also observed. Gene expression analysis revealed an upregulation of ESR1, ESR2, FSHR, and BAX, while AR and COX4l1 expression were downregulated. These findings offer new insights into the potential endocrine-disrupting and cytotoxic effects of CBZ, highlighting its potential role in compromising male reproductive health.

Melamine, a nitrogen-rich industrial chemical, has raised increasing concern as an emerging environmental contaminant with potential reproductive toxicity. While its nephrotoxic effects are well established, the direct impact of melamine on human sperm remains poorly defined. In this study, we investigated the in vitro effects of melamine on human sperm, under both capacitating and non-capacitating conditions. Functional analyses revealed that the exposure to 0.8 mM melamine, the highest non-cytotoxic concentration in vitro, significantly compromised sperm motility and disrupted key capacitation processes, including tyrosine phosphorylation patterns, cholesterol efflux, and the acrosome reaction. Molecular assessments demonstrated melamine-induced mitochondrial dysfunction, characterized by COX4I1 downregulation, reduced mitochondrial membrane potential, and altered reactive oxygen species production. In parallel, gene expression analyses revealed the activation of apoptotic pathways, with the upregulation of BAX and downregulation of BCL2, changes that were more pronounced during capacitation. Furthermore, melamine exposure significantly increased sperm DNA fragmentation and denaturation, indicating genotoxic stress. Collectively, these findings demonstrate that even low, non-cytotoxic concentrations of melamine compromise sperm function by disrupting capacitation, mitochondrial activity, and genomic integrity. This study identifies capacitation as a critical window of vulnerability and underscores the need to consider melamine as a potential environmental risk factor for male reproductive health.

Indigenous Saccharomyces cerevisiae strains persist within individual cellars and contribute to the character of spontaneously fermented wines, yet the molecular basis of their oenological differences remains largely unknown. In this work, eight predominant strains isolated from spontaneous fermentations in four Tuscan wineries were characterized by technological traits of oenological relevance and laboratory-scale fermentations of both a synthetic must and Sangiovese grape must. In each must, the strains reached similar final ethanol concentrations, although lag phase and maximum fermentation rate differed significantly, whereas the resulting wines differed significantly in glycerol, acetic acid and phenolic alcohol content and in the scores assigned by a trained sensory panel. Four strains spanning the observed range of behaviors were hence analyzed by two-dimensional polyacrylamide gel electrophoresis (2D-PAGE) and Matrix-Assisted Laser Desorption/Ionization Time-Of-Flight Mass Spectrometry (MALDI-ToF MS) at an early and a mid-fermentation stage, yielding 30–46 differentially abundant proteins identified per strain. Strain-specific abundance of proteins involved in glycerol formation, acetaldehyde oxidation and sulfur amino acid metabolism paralleled the corresponding differences in glycerol, acetic acid and hydrogen sulfide production, whereas glycolytic enzyme abundance varied without any corresponding variation in ethanol. Proteomic analysis thus identified candidate molecular correlates of the traits that distinguish strains, suggesting that it could complement traditional screening in the selection of indigenous starter cultures.