کد مقاله | کد نشریه | سال انتشار | مقاله انگلیسی | نسخه تمام متن |
---|---|---|---|---|
2149254 | 1089609 | 2007 | 15 صفحه PDF | دانلود رایگان |
عنوان انگلیسی مقاله ISI
Validation of a high-throughput in vitro alkaline elution/rat hepatocyte assay for DNA damage
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کلمات کلیدی
MDAdimethylnitrosaminessDNADETCAAFDMNBAPH3PO4BNFCASKH2PO4SLSdsDNAUS National Toxicology ProgramSDSNTP4,4′-MethylenedianilineN-Lauroyl sarcosine2-acetylaminofluorene - 2-استامینامین فلوئورنDMSO - DMSOSingle-stranded DNA - DNA تک رشته ایdouble-stranded DNA - DNA رشته ایβ-naphthoflavone - β-naftoflavoneAdenosine Triphosphate - آدنوزین تری فسفاتATP - آدنوزین تری فسفات یا ATPAutomation - اتوماسیونAlkaline elution - اسیدهای قلیاییValidation - اعتبار سنجیBenzo[a]pyrene - بنزو [a] پییرنGray - خاکستریChemical Abstracts Service - خدمات خلاصه شیمیDaba - دافاdiethyldithiocarbamate - دیاتیدیدیتیو کرباماتDimethyl sulfoxide - دیمتیل سولفواکسیدsodium dodecyl sulfate - سدیم دودسیل سولفاتSodium Phenobarbital - سدیم فنوباربیتالCytotoxicity - سمیت سلولیGenotoxicity - سمیت ژنتیکیPhosphoric acid - فسفریک اسیدHepatocyte - هپاتوسیتpotassium phosphate - پتاسیم فسفات
موضوعات مرتبط
علوم زیستی و بیوفناوری
بیوشیمی، ژنتیک و زیست شناسی مولکولی
تحقیقات سرطان
پیش نمایش صفحه اول مقاله
چکیده انگلیسی
In vitro alkaline elution is a sensitive and specific short term assay which measures DNA strand breakage in a mammalian test system (primary rat hepatocytes). This lab has previously demonstrated the performance of the assay with known genotoxic and non-genotoxic compounds. The methodology employed has relatively low sample throughput and is labor-intensive, requiring a great deal of manual processing of samples in a format that is not amenable to automation. Here, we present an automated version of the assay. This high-throughput alkaline elution assay (HT-AE) was made possible through 3 key developments: (1) DNA quantitation using PicoGreen® and OliGreen® fluorescent DNA binding dyes; (2) design and implementation of a custom automation system; and (3) reducing the assay to a 96-well plate format. The assay can now be run with 5-50Â mg of test compound. HT-AE was validated in a similar manner as the original assay, including assessment of non-genotoxic and non-carcinogenic compounds and evaluation of cytotoxicity to avoid confounding effects of toxicity-associated DNA degradation. The validation test results from compounds of known genotoxic potential were used to set appropriate criteria to classify alkaline elution results for genotoxicity.
ناشر
Database: Elsevier - ScienceDirect (ساینس دایرکت)
Journal: Mutation Research/Genetic Toxicology and Environmental Mutagenesis - Volume 629, Issue 1, 20 April 2007, Pages 49-63
Journal: Mutation Research/Genetic Toxicology and Environmental Mutagenesis - Volume 629, Issue 1, 20 April 2007, Pages 49-63
نویسندگان
Robert Gealy, Jennifer L. Wright-Bourque, Andrew R. Kraynak, Troy W. McKelvey, John E. Barnum, Richard D. Storer,