Glyphosate is the active ingredient of Roundup^®, which is one of the most popular herbicides worldwide. Although many studies have focused on the reproductive toxicity of glyphosate or glyphosate-based herbicides, the majority of them have concluded that the effect of the specific herbicide is negligible, while only a few studies indicate the male reproductive toxicity of glyphosate alone. The aim of the present study was to investigate the effect of 0.36 mg/L glyphosate on sperm motility and sperm DNA fragmentation (SDF). Thirty healthy men volunteered to undergo semen analysis for the purpose of the study. Sperm motility was calculated according to WHO 2010 guidelines at collection time (zero time) and 1 h post-treatment with glyphosate. Sperm DNA fragmentation was evaluated with Halosperm^® G2 kit for both the control and glyphosate-treated sperm samples. Sperm progressive motility of glyphosate-treated samples was significantly reduced after 1 h post-treatment in comparison to the respective controls, in contrast to the SDF of glyphosate-treated samples, which was comparable to the respective controls. Conclusively, under these in vitro conditions, at high concentrations that greatly exceed environmental exposures, glyphosate exerts toxic effects on sperm progressive motility but not on sperm DNA integrity, meaning that the toxic effect is limited only to motility, at least in the first hour.

Background: Only one study has reported the presence of extracellular vesicles (EVs) in COPD patients’ sputum. Thus, we aimed to isolate and characterize EVs from COPD and healthy individuals’ sputum. Methods: A total of 20 spontaneous sputum samples from COPD patients (m/f: 19/1) and induced sputum samples from healthy controls (m/f: 8/2) were used for EV isolation. The sputum supernatants were resuspended in PBS, precleared by centrifugation at 800× g for 10 min at 4 °C, and passed through a 0.22 μm filter (Millipore, Burlington, MA, USA). EVs were isolated by a standard membrane affinity spin column method (exoEasy maxi kit, Qiagen, Hilden, Germany). The EVs were then characterized by assessing their morphology and size using Transmission Electron Microscopy (TEM) and determining the CD9 and CD81 EV-markers with Western blot analysis. Results: The EVs had a spherical shape and their mean diameter in the COPD patients was significantly greater than in the controls. Enrichment of the EV markers, CD9 and CD81, were detected in both the healthy and COPD individuals. Total EV-associated protein was significantly increased in the COPD patients compared to the controls. ROC analysis showed that total EV-associated protein in the sputum could be used to differentiate between the controls and COPD patients, with a sensitivity of 80% and a specificity of 70% at a cut-off point of 55.59 μg/mL (AUC = 0.8150). Conclusions: EVs were detectable in both the COPD and healthy individuals’ sputum. The ratio of EVs in the 150–200 nm range was twice as high in the COPD patients than in the controls. The COPD patients’ sputum contained increased total EV-associated protein as compared to controls, highlighting their value as a new source of specific exoproteins.

Background: Chronic obstructive pulmonary disease (COPD) is a heterogeneous condition marked by airway inflammation, airflow limitation, and structural remodeling. Exosomal microRNAs (exo-miRNAs) are stable, cell-free biomarkers reflecting airway molecular changes. While serum and BALF exosomal miRNAs have been examined, sputum-derived profiles remain underexplored. Methods: Induced sputum was collected from 20 clinically stable COPD patients and 10 age-matched healthy controls. Exosomes were isolated by polymer-based precipitation and verified by transmission electron microscopy and Western blotting for CD9 and CD81. Nine candidate miRNAs (miR-21, miR-155, miR-34a, miR-126, miR-210, miR-146a, miR-199a-5p, miR-223, miR-1246) were quantified by RT-qPCR. Group comparisons used the Mann–Whitney U test, correlations Pearson’s r, and diagnostic accuracy ROC analysis. Results: Sputum-derived exosomes displayed characteristic morphology and canonical protein markers. COPD patients showed significant dysregulation of exosomal miRNAs, including upregulation of miR-21 (fold change = 3.4; 95% CI: 0.12–0.64 vs. 0.18–0.22; p < 0.001) and miR-223 (fold change = 2.1; 95% CI: 0.00–3.79 vs. 0.86–1.22; p = 0.004), and downregulation of miR-155 (fold change = 0.35; 95% CI: 0.43–0.67 vs. 0.86–1.22; p = 0.002), miR-126 (fold change = 0.42; 95% CI: 0.30–0.39 vs. 0.80–1.42; p = 0.009), and miR-146a (fold change = 0.28; 95% CI: 0.49–1.12 vs. 0.87–1.35; p = 0.006). miR-21 correlated with symptom burden (CAT; r = 0.445; p = 0.049). Among individual biomarkers, miR-155 exhibited the best diagnostic performance for COPD detection (AUC = 0.730; 95% CI: 0.53–0.93), which further improved when combined with miR-126 and miR-146a (AUC = 0.841; 95% CI: 0.69–0.98). For disease stratification, miR-126 most effectively discriminated mild from moderate-to-severe COPD (AUC = 0.728; 95% CI: 0.50–0.96). These results indicate that sputum-derived exosomal miRNAs—particularly miR-155, miR-126, and miR-146a—may serve as promising non-invasive biomarkers for COPD diagnosis and clinical phenotyping. Conclusions: Sputum exosomal miRNAs reveal a distinct COPD-specific signature reflecting inflammation, impaired repair, and immune dysregulation. Composite panels incorporating miR-155, miR-126, and miR-146a enhance diagnostic accuracy and could be integrated into non-invasive workflows for COPD detection and staging.

Background: N-terminal pro-B-type natriuretic peptide (NT-proBNP) is released in response to increased cardiac wall stress and is used as a biomarker for volume overload and heart failure (HF). It is also elevated in atrial fibrillation (AF) and inflammation. However, in hemodialysis (HD) patients, its interpretation is complicated by reduced renal clearance, large fluid shifts between dialysis sessions, and chronic inflammation. Methods: In 123 HD patients, we examined the relationship between NT-proBNP and interdialytic weight gain, HF with preserved ejection fraction (HFpEF), left ventricular systolic dysfunction (LVSD), and AF, as well as the impact of inflammation. Clinical characteristics, laboratory data, and echocardiography (within three months) were evaluated, while serum NT-proBNP and calprotectin levels were measured by ELISA. Results: NT-proBNP showed no association with interdialytic weight gain and did not identify HFpEF. Inflammatory markers (C-reactive protein and calprotectin) correlated positively with NT-proBNP. Multivariable analysis demonstrated that LVSD, AF, and inflammation remained independent predictors of NT-proBNP levels. Although NT-proBNP levels were higher in LVSD, its diagnostic performance was poor (AUC 0.627). In contrast, NT-proBNP was significantly elevated in patients with AF and showed good diagnostic performance (AUC 0.801). Conclusions: In HD patients, NT-proBNP is not correlated with interdialytic weight gain, performs poorly as a marker of LVSD, but may serve as a useful marker of AF.