{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,1,2]],"date-time":"2026-01-02T17:20:52Z","timestamp":1767374452479},"reference-count":30,"publisher":"Wiley","issue":"3","license":[{"start":{"date-parts":[[2006,7,14]],"date-time":"2006-07-14T00:00:00Z","timestamp":1152835200000},"content-version":"vor","delay-in-days":6403,"URL":"http:\/\/onlinelibrary.wiley.com\/termsAndConditions#vor"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Environ and Mol Mutagen"],"published-print":{"date-parts":[[1989,1]]},"abstract":"<jats:title>Abstract<\/jats:title><jats:p>Alkaline elution is a sensitive and commonly used technique to detect cellular DNA damage in the form of DNA strand breaks and DNA cross\u2010links. Conventional alkaline elution procedures have extensive equipment requirements and are tedious to perform. Our laboratory recently presented a rapid, simplified, and sensitive modification of the alkaline elution technique to detect carcinogen\u2010induced DNA strand breaks. In the present study, we have further modified this technique to enable the rapid characterization of chemically induced DNA\u2010interstrand and DNA\u2010protein associated cross\u2010links in cultured epithelial cells. Cells were exposed to three known DNA cross\u2010linking agents, nitrogen mustard (HN2), mitomycin C (MMC), or ultraviolet irradiation (UV). One hour exposures of HN2 at 0.25, 1.0, and 4.0 \u03bcM or of MMC at 20, 40, and 60 \u03bcM produced a dose\u2010dependent induction of total DNA cross\u2010links by these agents. Digestion with proteinase K revealed that HN2 and MMC induced both DNA\u2010protein cross\u2010links and DNA\u2010interstrand cross\u2010links. Ultraviolet irradiation induced both DNA cross\u2010links and DNA strand breaks, the latter of which were either protein and nonprotein associated. The results demonstrate that gravity\u2010flow alkaline elution is a sensitive and accurate method to characterize the molecular events of DNA cross\u2010linking. Using this procedure, elution of DNA from treated cells is completed in 1 hr, and only three fractions per sample are analyzed. This method may be useful as a rapid screening assay for genotoxicity and\/or as an adjunct to other predictive assays for potential mutagenic or carcinogenic agents.<\/jats:p>","DOI":"10.1002\/em.2850130304","type":"journal-article","created":{"date-parts":[[2007,2,21]],"date-time":"2007-02-21T16:39:59Z","timestamp":1172075999000},"page":"211-217","source":"Crossref","is-referenced-by-count":13,"title":["Rapid detection of DNA\u2010interstrand and DNA\u2010protein cross\u2010links in mammalian cells by gravity\u2010flow alkaline elution"],"prefix":"10.1002","volume":"13","author":[{"given":"Jeffrey R.","family":"Hincks","sequence":"first","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"suffix":"Jr.","given":"Roger A.","family":"Coulombe","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"311","published-online":{"date-parts":[[2006,7,14]]},"reference":[{"key":"e_1_2_1_2_1","first-page":"3510","article-title":"Interactions of mitomycin C with mammalian DNA detected by alkaline elution","volume":"45","author":"Dorr RT","year":"1985","journal-title":"Cancer Res"},{"key":"e_1_2_1_3_1","volume-title":"Statistics For Research","author":"Dowdy S","year":"1983"},{"key":"e_1_2_1_4_1","doi-asserted-by":"publisher","DOI":"10.1111\/j.1751-1097.1978.tb07669.x"},{"key":"e_1_2_1_5_1","first-page":"2114","article-title":"DNA damage and repair in mouse leukemia L1210 cells treated with nitrogen mustard, 1, 3\u2010Bis(2\u2010chloroethyl)\u20101\u2010nitrosourea, and other nitrosoureas","volume":"37","author":"Ewig RAG","year":"1977","journal-title":"Cancer Res"},{"key":"e_1_2_1_6_1","doi-asserted-by":"publisher","DOI":"10.1016\/0005-2787(76)90196-9"},{"key":"e_1_2_1_7_1","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.73.1.39"},{"key":"e_1_2_1_8_1","doi-asserted-by":"publisher","DOI":"10.1016\/0005-2787(77)90253-2"},{"key":"e_1_2_1_9_1","first-page":"704","article-title":"DNA\u2010protein cross\u2010linking by chemical carcinogens in mammalian cells","volume":"39","author":"Fornace AJ","year":"1979","journal-title":"Cancer Res"},{"key":"e_1_2_1_10_1","first-page":"145","article-title":"Detection of DNA single\u2010strand breaks produced during the repair of damage by DNA\u2010protein cross\u2010linking agents","volume":"42","author":"Fornace AJ","year":"1982","journal-title":"Cancer Res"},{"key":"e_1_2_1_11_1","doi-asserted-by":"publisher","DOI":"10.1016\/0006-291X(86)90325-6"},{"key":"e_1_2_1_12_1","unstructured":"ICRU(1969):Report 14. 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