{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,5,4]],"date-time":"2026-05-04T05:21:16Z","timestamp":1777872076105,"version":"3.51.4"},"reference-count":36,"publisher":"Wiley","issue":"5","license":[{"start":{"date-parts":[[2014,7,18]],"date-time":"2014-07-18T00:00:00Z","timestamp":1405641600000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/onlinelibrary.wiley.com\/termsAndConditions#vor"}],"funder":[{"name":"Bijzonder OnderzoeksFonds Uhasselt","award":["BOF08G01"],"award-info":[{"award-number":["BOF08G01"]}]},{"DOI":"10.13039\/501100001659","name":"Deutsche Forschungsgemeinschaft","doi-asserted-by":"crossref","award":["TH345\/11-1"],"award-info":[{"award-number":["TH345\/11-1"]}],"id":[{"id":"10.13039\/501100001659","id-type":"DOI","asserted-by":"crossref"}]},{"name":"Methusalem","award":["08M03VGRJ"],"award-info":[{"award-number":["08M03VGRJ"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["J of Applied Toxicology"],"published-print":{"date-parts":[[2015,5]]},"abstract":"<jats:title>Abstract<\/jats:title><jats:p>Cadmium (Cd) is a toxic metal with a long half\u2010life in biological systems. This half\u2010life is partly as a result of metallothioneins (MTs), metal\u2010binding proteins with a high affinity for Cd. The high retention properties of the kidneys reside in proximal tubular cells that possess transport mechanisms for Cd\u2010MT uptake, ultimately leading to more Cd accumulation. Researchers have studied MT\u2013metal interactions using various techniques including quantitative real\u2010time PCR (qPCR), an efficient tool for quantifying gene expression. Often a poor choice of reference genes, which is represented by their instability and condition dependency, leads to inefficient normalization of gene expression data and misinterpretations. This study demonstrates the importance of an efficient normalization strategy in toxicological research. A selection of stable reference genes was proposed in order to acquire reliable and reproducible gene quantification under metal stress using MT expression as an example. Moreover, <jats:italic>in vitro<\/jats:italic> and <jats:italic>in vivo<\/jats:italic> setups were compared to identify the influence of toxicological compounds in function of the experimental design. This study shows that glyceraldehyde\u20103\u2010phosphate dehydrogenase (Gapdh), tyrosine monooxygenase\/tryptophan5\u2010monooxygenase activation\u2010protein, zeta polypeptide (Ywhaz) and beta\u2010actin (Actb) are the most stable reference genes in a kidney proximal tubular cell line exposed to moderate and high Cd concentrations, applied as CdCl<jats:sub>2<\/jats:sub>. A slightly different sequence in reference gene stability was found in renal cells isolated from rats <jats:italic>in vivo<\/jats:italic> exposed to Cd. It was further shown that three reference genes are required for efficient normalization in this experimental setup. This study demonstrates the importance of an efficient normalization strategy in toxicological research. Copyright \u00a9 2014 John Wiley &amp; Sons, Ltd.<\/jats:p>","DOI":"10.1002\/jat.3047","type":"journal-article","created":{"date-parts":[[2014,7,18]],"date-time":"2014-07-18T05:46:52Z","timestamp":1405662412000},"page":"478-484","source":"Crossref","is-referenced-by-count":16,"title":["Renal cells exposed to cadmium <i>in vitro<\/i> and <i>in vivo<\/i>: normalizing gene expression data"],"prefix":"10.1002","volume":"35","author":[{"given":"A. R.","family":"Nair","sequence":"first","affiliation":[{"name":"Centre for Environmental Sciences Hasselt University  Agoralaan Building D 3590 Diepenbeek Belgium"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"K.","family":"Smeets","sequence":"additional","affiliation":[{"name":"Centre for Environmental Sciences Hasselt University  Agoralaan Building D 3590 Diepenbeek Belgium"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"E.","family":"Keunen","sequence":"additional","affiliation":[{"name":"Centre for Environmental Sciences Hasselt University  Agoralaan Building D 3590 Diepenbeek Belgium"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"W.\u2010K.","family":"Lee","sequence":"additional","affiliation":[{"name":"Pathophysiology &amp; Toxicology, Centre for Biomedical Education and Research Witten\/Herdecke University  Germany"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"F.","family":"Th\u00e9venod","sequence":"additional","affiliation":[{"name":"Pathophysiology &amp; Toxicology, Centre for Biomedical Education and Research Witten\/Herdecke University  Germany"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"E.","family":"Van Kerkhove","sequence":"additional","affiliation":[{"name":"Centre for Environmental Sciences Hasselt University  Agoralaan Building D 3590 Diepenbeek Belgium"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"A.","family":"Cuypers","sequence":"additional","affiliation":[{"name":"Centre for Environmental Sciences Hasselt University  Agoralaan Building D 3590 Diepenbeek Belgium"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"311","published-online":{"date-parts":[[2014,7,18]]},"reference":[{"key":"e_1_2_7_2_1","doi-asserted-by":"publisher","DOI":"10.1158\/0008-5472.CAN-04-0496"},{"key":"e_1_2_7_3_1","volume-title":"Cadmium toxicity\u2010Case Studies in Environmental Medicine","author":"ATSDR","year":"2008"},{"key":"e_1_2_7_4_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.taap.2004.09.007"},{"key":"e_1_2_7_5_1","doi-asserted-by":"publisher","DOI":"10.1373\/clinchem.2008.112797"},{"key":"e_1_2_7_6_1","doi-asserted-by":"publisher","DOI":"10.1038\/nmeth.2697"},{"key":"e_1_2_7_7_1","doi-asserted-by":"publisher","DOI":"10.1007\/s10534-010-9329-x"},{"key":"e_1_2_7_8_1","doi-asserted-by":"publisher","DOI":"10.1186\/1471-2199-10-54"},{"key":"e_1_2_7_9_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.tiv.2010.08.007"},{"key":"e_1_2_7_10_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.envres.2006.05.009"},{"key":"e_1_2_7_11_1","volume-title":"Beryllium, Cadmium, Mercury, and Exposures in the Glass Manufacturing Industry","author":"IARC","year":"1993"},{"key":"e_1_2_7_12_1","doi-asserted-by":"publisher","DOI":"10.1007\/s10529-011-0552-5"},{"issue":"1","key":"e_1_2_7_13_1","doi-asserted-by":"crossref","first-page":"1","DOI":"10.5271\/sjweh.270","volume":"24","author":"J\u00e4rup L","year":"1998","journal-title":"Scand. J. Work Environ. Health"},{"key":"e_1_2_7_14_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.taap.2009.01.011"},{"key":"e_1_2_7_15_1","doi-asserted-by":"publisher","DOI":"10.1146\/annurev.pharmtox.39.1.267"},{"key":"e_1_2_7_16_1","doi-asserted-by":"publisher","DOI":"10.1179\/1743132812Y.0000000060"},{"key":"e_1_2_7_17_1","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.090091397"},{"key":"e_1_2_7_18_1","doi-asserted-by":"publisher","DOI":"10.1152\/ajpcell.00197.2007"},{"key":"e_1_2_7_19_1","doi-asserted-by":"publisher","DOI":"10.1379\/1466-1268(2001)006<0097:MGEIPL>2.0.CO;2"},{"key":"e_1_2_7_20_1","doi-asserted-by":"publisher","DOI":"10.3390\/ijms14036116"},{"key":"e_1_2_7_21_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.jneumeth.2009.12.018"},{"key":"e_1_2_7_22_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.taap.2009.03.015"},{"key":"e_1_2_7_23_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.biochi.2009.06.014"},{"key":"e_1_2_7_24_1","doi-asserted-by":"publisher","DOI":"10.1007\/978-3-0348-6784-9_4"},{"key":"e_1_2_7_25_1","doi-asserted-by":"publisher","DOI":"10.1007\/s10646-011-0809-8"},{"key":"e_1_2_7_26_1","doi-asserted-by":"publisher","DOI":"10.1124\/jpet.110.166769"},{"key":"e_1_2_7_27_1","doi-asserted-by":"publisher","DOI":"10.1007\/s00425-008-0706-4"},{"key":"e_1_2_7_28_1","doi-asserted-by":"publisher","DOI":"10.1097\/01.HP.0000338421.07312.ed"},{"key":"e_1_2_7_29_1","doi-asserted-by":"publisher","DOI":"10.1006\/mcpr.2001.0376"},{"key":"e_1_2_7_30_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.cbpb.2008.07.010"},{"key":"e_1_2_7_31_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.taap.2009.01.013"},{"key":"e_1_2_7_32_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.tox.2007.03.022"},{"key":"e_1_2_7_33_1","doi-asserted-by":"publisher","DOI":"10.1186\/gb-2002-3-7-research0034"},{"key":"e_1_2_7_34_1","doi-asserted-by":"publisher","DOI":"10.1007\/s11033-011-0897-9"},{"key":"e_1_2_7_35_1","doi-asserted-by":"publisher","DOI":"10.1038\/ki.1996.295"},{"key":"e_1_2_7_36_1","doi-asserted-by":"publisher","DOI":"10.1006\/bbrc.1999.0815"},{"key":"e_1_2_7_37_1","doi-asserted-by":"publisher","DOI":"10.1093\/toxsci\/kfm260"}],"container-title":["Journal of Applied Toxicology"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/api.wiley.com\/onlinelibrary\/tdm\/v1\/articles\/10.1002%2Fjat.3047","content-type":"unspecified","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/analyticalsciencejournals.onlinelibrary.wiley.com\/doi\/pdf\/10.1002\/jat.3047","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,15]],"date-time":"2025-10-15T15:34:03Z","timestamp":1760542443000},"score":1,"resource":{"primary":{"URL":"https:\/\/analyticalsciencejournals.onlinelibrary.wiley.com\/doi\/10.1002\/jat.3047"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2014,7,18]]},"references-count":36,"journal-issue":{"issue":"5","published-print":{"date-parts":[[2015,5]]}},"alternative-id":["10.1002\/jat.3047"],"URL":"https:\/\/doi.org\/10.1002\/jat.3047","archive":["Portico"],"relation":{},"ISSN":["0260-437X","1099-1263"],"issn-type":[{"value":"0260-437X","type":"print"},{"value":"1099-1263","type":"electronic"}],"subject":[],"published":{"date-parts":[[2014,7,18]]}}}