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Determination of double-and single-stranded DNA breaks in bovine sperm is predictive of their fertilizing capacity

Journal of Animal Science and Biotechnology 2023年第1期14卷 134-151页

作者： Jordi ribas‑Maynou Ariadna Delgado‑Bermúdez Yentel Mateo‑Otero Estel Vinolas Carlos O.Hidalgo WSteven Ward marc Yeste Biotechnology of Animal and Human Reproduction(TechnoSperm) Faculty of SciencesInstitute of Food and Agricultural TechnologyUniversity of GironaC/Maria Aurèlia Campany69ES‑17003 GironaSpain. Unit of Cell Biology Department of BiologyFaculty of SciencesUniversity of GironaES‑17003 GironaSpain. Institute for Biogenesis Research Department of AnatomyBiochemistry&PhysiologyJohn A.Burns School of MedicineUniversity of Hawaii at ManoaHonoluluHI 96822USA. Department of Animal Selection and Reproduction The Regional Agri-Food Research and Development Service of Asturias(SERIDA)ES‑33394 GijónSpain. Catalan Institution for Research and Advanced Studies(ICREA) ES‑08010 BarcelonaSpain.

Background:The analysis of chromatin integrity has become an important determinant of sperm *** frozenthawed bovine sperm,neither the sequence of post-thaw injury events nor the dynamics of different types of sperm DNA breaks are well *** aim of the present work was to describe such sperm degradation aftermath focusing on DNA damage dynamics,and to assess if this parameter can predict pregnancy rates in ***:A total of 75 cryopreserved ejaculates from 25 Holstein bulls were evaluated at two post-thawing periods(0-2 h and 2-4 h),analyzing global and double-stranded DNA damage through alkaline and neutral Comet assays,chromatin deprotamination and decondensation,sperm motility,viability,acros.mal status,and intracellular levels of total ros.superoxides and *** of 59,605 females was conducted using sperm from the same bulls,thus obtaining the non-return to estrus rates after 90 d(NRR).Results showed an increased rate of double-stranded breaks in the first period(0-2 h:1.29±1.01%/h vs.2-4 h:0.13±1.37%/h;P0.05).Finally,whereas no correlations between bull fertility and any dynamic parameter were found,there were correlations between the NRR and the basal percentage of highly-damaged sperm assessed with the alkaline Comet(Rs=-0.563,P=0.003),between NRR and basal progressive motility(Rs=0.511,P=0.009),and between NRR and sperm with high ros.at 4 h post-thaw(Rs=0.564,P=0.003).Conclusion:The statistically significant correlations found between intracellular ros.sperm viability,sperm motility,DNA damage

关键词： Cattle Chromatin Comet test DNA damage Fertility Sperm Sperm quality

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Red Onion (Allium Cepa L.) Methanolic Extract Increases Extracellular Nucleotide Hydrolysis in Rat Serum

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Open Journal of Applied Sciences 2020年第12期10卷 864-876页

作者： Janayne Luihan Silva Bruno Quintanilha Faria Vinícius marques Arruda Fernanda Cardoso da Silva Joyce Ferreira da Costa Guerra Cristina ribas Fürstenau Instituto de Ciências Biológicas Universidade Federal de Minas Gerais Belo Horizonte Brazil Curso de Biotecnologia Universidade Federal de Uberlandia Patos de Minas Brazil Programa de Pós-graduaç o em Biotecnologia Instituto de Biotecnologia Universidade Federal de Uberlandia Patos de Minas Brazil Centro de Ciências Naturais e Humanas Universidade Federal do ABC Santo André Brazil

Ectonucleotidases are enzymes involved in nucleotides metabolism. The amount of circulating nucleotides may modulate distinct pathophysiological processes in the vasculature, including platelet aggregation and vascular tone. Onion (Allium cepa L.) phenolic compounds modulate enzymatic activity. The present study evaluated the total phenolic content of red onion methanolic extract, its antioxidant capacity, and its ability to interfere in nucleotides hydrolyses in rat serum. Total phenolic content was determined with the Folin-Ciocalteau reagent using gallic acid as a standard, while total flavonoid content was obtained through the aluminum chloride colorimetric method with quercetin as a standard. Antioxidant capacity was evaluated from the ability of the extract to scavenge ABTS·+ and DPPH· radicals. ATP, ADP, AMP, and p-Nph-5'-TMP hydrolyses were colorimetrically determined in response to different onion extract concentrations (0, 125, 250, 500, or 1000 μg/mL). Phenolic content of the extract was 54.35 mg GAE 100 g-1 sample, while flavonoid content was 7.22 mg quercetin g-1 sample. The IC50 value for ABTS·+ was 374.13 ± 7.52 μg/mL, while it was 440.29 ± 15.17 μg/mL for DPPH·. Red onion extract increased ADP and p-Nph-5'-TMP hydrolysis. The results confirmed that red onion contains high content of antioxidant, mainly flavonoids, and high antioxidant capacity. Additionally, biochemical studies suggest that the increased ADP breakdown may be important to regulate vascular processes. As it occurs for other enzymes, the antioxidant capacity of onion extract may neutralize reactive oxygen species (ros. formation and favor ectonucleotidase activity and the hydrolysis of ADP, a major platelet agonist.

关键词： Red Onion Flavonoids Reactive Oxygen Species Ectonucleotidases Vascular Homeostasis

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