{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2025,2,21]],"date-time":"2025-02-21T10:53:30Z","timestamp":1740135210904,"version":"3.37.3"},"reference-count":16,"publisher":"Springer Science and Business Media LLC","issue":"1","license":[{"start":{"date-parts":[[2022,11,25]],"date-time":"2022-11-25T00:00:00Z","timestamp":1669334400000},"content-version":"tdm","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0"},{"start":{"date-parts":[[2022,11,25]],"date-time":"2022-11-25T00:00:00Z","timestamp":1669334400000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0"}],"funder":[{"DOI":"10.13039\/100009633","name":"Eunice Kennedy Shriver National Institute of Child Health and Human Development","doi-asserted-by":"publisher","award":["[R01HD096209]","[R01HD096209]","[R01HD096209]"],"award-info":[{"award-number":["[R01HD096209]","[R01HD096209]","[R01HD096209]"]}],"id":[{"id":"10.13039\/100009633","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["link.springer.com"],"crossmark-restriction":false},"short-container-title":["BMC Biotechnol"],"abstract":"<jats:title>Abstract<\/jats:title><jats:sec>\n                <jats:title>Background<\/jats:title>\n                <jats:p>A mixture of phenol and guanidine isothiocyanate (\u201cP\/GI\u201d, the principal components of TRIzol\u2122 and similar products) is routinely used to isolate RNA, DNA, and proteins from a single specimen. In time-course experiments of cells grown in tissue culture, replicate wells are often harvested sequentially and compared, with the assumption that in-well lysis and complete aspiration of P\/GI has no effect on continuing cultures in nearby wells.<\/jats:p>\n              <\/jats:sec><jats:sec>\n                <jats:title>Methods<\/jats:title>\n                <jats:p>To test this assumption, we investigated morphology and function of RAW 264.7 cells (an immortalized mouse macrophage cell line) cultured in covered 96-well plates for 4, 8, or 24\u00a0h at varying distances from a single control well or a well into which P\/GI had been deposited and immediately aspirated completely.<\/jats:p>\n              <\/jats:sec><jats:sec>\n                <jats:title>Results<\/jats:title>\n                <jats:p>Time- and distance-dependent disruptions resulting from proximity to a single well containing trace residual P\/GI were seen in cell morphology (blebbing, cytoplasmic disruption, and accumulation of intracellular vesicles), cell function (pH of culture medium), and expression of genes related to inflammation (<jats:italic>Tnf\u03b1<\/jats:italic>) and autophagy (<jats:italic>Lc3b<\/jats:italic>). There was no transcriptional change in the anti-apoptotic gene <jats:italic>Mcl1<\/jats:italic>, nor the pro-apoptotic gene <jats:italic>Hrk<\/jats:italic>, nor in P\/GI-unexposed control cultures. LPS-stimulated cells incubated near P\/GI had lower expression of the cytokine <jats:italic>Il6<\/jats:italic>. These effects were seen as early as 4\u00a0h of exposure and at a distance of up to 3 well units from the P\/GI-exposed well.<\/jats:p>\n              <\/jats:sec><jats:sec>\n                <jats:title>Conclusions<\/jats:title>\n                <jats:p>Exposure to trace residual quantities of P\/GI in covered tissue culture plates leads to substantial disruption of cell morphology and function in as little as 4\u00a0h, possibly through induction of autophagy but not apoptosis. This phenomenon should be considered when planning time-course experiments in multi-well covered tissue culture plates.<\/jats:p>\n              <\/jats:sec>","DOI":"10.1186\/s12896-022-00766-2","type":"journal-article","created":{"date-parts":[[2022,11,25]],"date-time":"2022-11-25T21:47:36Z","timestamp":1669412856000},"update-policy":"https:\/\/doi.org\/10.1007\/springer_crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["Toxic effects of trace phenol\/guanidine isothiocyanate (P\/GI) on cells cultured nearby in covered 96-well plates"],"prefix":"10.1186","volume":"22","author":[{"given":"Madeline","family":"Snedden","sequence":"first","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Lavisha","family":"Singh","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Chandrashekara","family":"Kyathanahalli","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Emmet","family":"Hirsch","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"297","published-online":{"date-parts":[[2022,11,25]]},"reference":[{"key":"766_CR1","doi-asserted-by":"publisher","first-page":"pdb.prot5439","DOI":"10.1101\/pdb.prot5439","volume":"2010","author":"DC Rio","year":"2010","unstructured":"Rio DC, Ares M, Hannon GJ, Nilsen TW. 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