Background: Male infertility is increasingly becoming a health and demographic problem. While it may originate from congenital or acquired diseases, it can also result from environmental exposure. Hence, the complexity of involved molecular mechanisms often requires a multiparametric approach. This study aimed to associate semen parameters with sperm DNA fragmentation, chromatin maturity and seminal plasma protein N-glycosylation. Methods: The study was conducted with 166 participants, 20–55 y old, 82 normozoospermic and 84 with pathological diagnosis. Sperm was analyzed by Halosperm assay and aniline blue staining, while seminal plasma total protein N-glycans were analyzed by ultra-high-performance liquid chromatography. Results: Sperm DNA fragmentation was significantly increased in the pathological group and was inversely correlated with sperm motility and viability. Seminal plasma total protein N-glycans were chromatographically separated in 37 individual peaks. The pattern of seminal plasma N-glycan peaks (SPGP) showed that SPGP14 significantly differs between men with normal and pathological semen parameters (p < 0.001). The multivariate analysis showed that when sperm chromatin maturity increases by 10%, SPGP17 decreases by 14% while SPGP25 increases by 25%. Conclusion: DNA integrity and seminal plasma N-glycans are associated with pathological sperm parameters. Specific N-glycans are also associated with sperm chromatin maturity and have a potential in future fertility research and clinical diagnostics.
Symbiosis is an evolutionary strategy that provides biological advantages. The most complex cooperation between symbiotic organisms is endosymbiosis, as it is the symbiosis between green hydra (Hydra viridissima Pallas, 1766) and unicellular photoautotrophic alga. We aimed to evaluate whether this symbiotic cooperation bestows higher resistance to stressors, in terms of preservation of DNA integrity, compared to free-living brown hydra (Hydra oligactis Pallas, 1766). Hydras were exposed to herbicide norflurazon at 2×10− 7 or 2×10− 6 mol/L and UV-B light of 254 nm, 0.023 mW/cm separately or simultaneously. By alkaline comet assay the level of primary DNA damage and by fluorescent staining apoptosis and necrosis were determined. Norflurazon at 2×10− 6 mol/L significantly increased comet assay descriptors in brown hydra compared to negative control (6.17 ± 0.6µm and 5.2 ± 1.7% vs. 2.9 ± 0.2µm and 1.2 ± 0.2%). The frequency of apoptotic and necrotic cells was significantly elevated either, being higher in brown hydra (25.7 ± 3.5% and 8.2 ± 0.2%) than green hydra (20.3 ± 2.5% and 2.8 ± 0.2%). UV-B irradiation induced significant DNA damage in brown hydra only (13.5 ± 1.0µm, 4.1 ± 1.0%). Simultaneous exposure to UV-B light and norflurazon led to synergistic increase in intensity of adverse effects. Norflurazon at 2×10− 7 mol/L combined with UV-B light induced significantly increased DNA lesion-level in brown hydra (15.8 ± 3.1%) and at concentration of 2×10− 6 mol/L in both species (brown: 15.0 ± 2.6%, green: 22.0 ± 3.0%). Cytotoxicity and increased hedgehog nucleoids frequency were recorded, significantly more pronounced in brown (78.3 ± 9.4% and 56.4 ± 6.0%) compared to green hydra (34.7 ± 2.5% and 24.2 ± 0.6%). We proved that evolutionary established symbiotic cooperation may contribute to higher resistance to cyto/genotoxic stressors.
Symbiosis is an evolutionary strategy that may provide biological advantages. The most complex and tight cooperation between symbiotic organisms is achieved in endosymbiosis, as it is the symbiosis between green hydra (Hydra viridissima Pallas, 1766) and unicellular photoautotrophic alga. We aimed to evaluate whether this symbiotic cooperation bestows higher resistance to environmental stressors of different nature, in terms of higher preservation of DNA integrity, compared to free-living brown hydra (Hydra oligactis Pallas, 1766). Hydras were exposed to herbicide norflurazon at of 2×10 or 2×10 mol/L and UV-B light of 254 nm, 0.023 mW/cm separately or simultaneously to detect additive or synergistic effect. We used alkaline comet assay to determine the level of primary DNA damage and fluorescent staining to detect apoptosis and necrosis. Norflurazon at 2×10-6 mol/L significantly increased both comet assay descriptors in brown hydra compared to negative control (6.17±0.6µm and 5.2±1.7% vs. 2.9 ± 0.2µm and 1.2 ± 0.2 %). The frequency of apoptotic and necrotic cells was significantly elevated either, being higher in brown hydra (25.7±3.5% and 8.2±0.2%) than green hydra (20.3±2.5% and 2.8±0.2%). UV-B irradiation induced significant DNA damage in brown hydra only (13.5±1.0µm, 4.1±1.0%). Simultaneous exposure to UV-B light and norflurazon led to synergistic increase in intensity of adverse effects. A concentration of 2×10 mol norflurazon/L combined with UV-B light induced significantly increased DNA lesion-level in brown hydra (15.8 ± 3.1%) and at concentration of 2×10 mol/L in both species (brown: 15.0±2.6%, green: 22.0±3.0%). Significant cytotoxicity and increased hedgehog nucleoids frequency were recorded, significantly more pronounced in brown (78.3±9.4% and 56.4±6.0%) compared to green hydra (34.7±2.5% and 24.2±0.6%). Besides genotoxicity, comet assay descriptors were significantly affected by and reflected cytotoxicity of norflurazon and/or UV-B light. We proved that evolutionary established symbiotic cooperation may contribute to higher resistance to cyto/genotoxic stressors.
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