{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,6]],"date-time":"2026-10-06T17:54:34Z","timestamp":1791309274073,"version":"4.3.1"},"reference-count":0,"publisher":"GN1 Sistemas e Publicacoes Ltd.","content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["JBRA"],"published-print":{"date-parts":[[2026]]},"abstract":"<jats:p>Objective: Density gradient centrifugation is widely used for sperm preparation, although centrifugation-induced oxidative stress may cause DNA damage. This study compared the rates of \u03b3H2AX-positive spermatozoa (a surrogate marker associated with DNA double-strand break\u2013related damage signaling) among sperm prepared by density gradient centrifugation, ZyM\u014dt, and Felix, and evaluated sperm recovery, total motility, and processing time.\nMethods: Fifteen fresh semen samples collected from January to June 2025 were processed in parallel using density gradient centrifugation, ZyM\u014dt, and Felix. Following sperm preparation, \u03b3H2AX immunostaining was performed, and at least 200 spermatozoa per sample were scored to determine the \u03b3H2AX-positive rate. Sperm recovery, total motility, and workflow-defined processing time were also recorded. Statistical analyses were performed using the Friedman test followed by Wilcoxon signed-rank tests with Bonferroni correction, and data were expressed as medians with interquartile ranges.\nResults: The \u03b3H2AX-positive rate was significantly lower in the ZyM\u014dt (11.8%) and Felix (10.0%) groups compared with the density gradient centrifugation group (16.0%; p&lt;0.01). Density gradient centrifugation yielded the highest sperm recovery, ZyM\u014dt achieved the highest total motility, and Felix required the shortest processing time, indicating that the three methods exhibit distinct performance characteristics.\nConclusion: The non-centrifugal systems ZyM\u014dt and Felix reduced the proportion of \u03b3H2AX-positive spermatozoa compared with density gradient centrifugation under our laboratory conditions while maintaining comparable overall performance. Whether these laboratory differences translate into improved assisted reproductive technology outcomes should be evaluated in future prospective studies.<\/jats:p>","DOI":"10.5935\/1518-0557.20260078","type":"journal-article","created":{"date-parts":[[2026,10,6]],"date-time":"2026-10-06T17:33:15Z","timestamp":1791307995000},"page":"1-9","source":"Crossref","is-referenced-by-count":0,"title":["Evaluation of DNA Double-Strand Breaks in Human Sperm Following Selection by Density Gradient Centrifugation, ZyM\u014dt, and Felix Techniques"],"prefix":"10.5935","volume":"30","author":[{"given":"Mitsuru","family":"Nago","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Akari","family":"Saito","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yuria","family":"Takahashi","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Eri","family":"Kamioka","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ami","family":"Fujisawa","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Mayuri","family":"Chino","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Manon","family":"Koyata","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yuta","family":"Kawagoe","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Bunpei","family":"Ishizuka","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"3948","published-online":{"date-parts":[[2026]]},"container-title":["JBRA Assisted Reproduction"],"original-title":[],"link":[{"URL":"https:\/\/www.jbra.com.br\/trab\/pub\/download_trabalho.php?fileSource=\/var\/www\/vhosts\/jbra.com.br\/media\/trab\/arq_8826&fileName=3545.pdf&id_trabalho=3323","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,10,6]],"date-time":"2026-10-06T17:33:23Z","timestamp":1791308003000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.jbra.com.br\/trab\/pub\/download_trabalho.php?fileSource=\/var\/www\/vhosts\/jbra.com.br\/media\/trab\/arq_8826&fileName=3545.pdf&id_trabalho=3323"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2026]]},"references-count":0,"URL":"https:\/\/doi.org\/10.5935\/1518-0557.20260078","relation":{},"ISSN":["1518-0557"],"issn-type":[{"value":"1518-0557","type":"print"}],"subject":[],"published":{"date-parts":[[2026]]}}}