A microgel electrophoresis technique for the direct quantitation of DNA damage and repair in individual fibroblasts cultured on microscope slides

Singh, Narendra P.; Tice, Raymond R.; Stephens, R. E.; Schneider, Edward L.

doi:10.1016/0165-1161(91)90008-v

Cited by 233 publications

(76 citation statements)

References 11 publications

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“…Presumably it is undetected by nick translation because the assay Table I DNA requires clean 3'-OH ends for polymerase activity. Hydrogen peroxide is a potent DNA strand-breaking agent, at very low doses (C 100 PM) in the comet assay [30], which should detect any type of nicking of supercoiled domains in the residual nuclear structure of the lysed cell [31]. "Dirty' strand breaks may indeed be detected more readily in the comet assay because they are less rapidly repaired.…”

Section: Discussionmentioning

confidence: 99%

Endogenous nitric oxide induced by interleukin‐1β in rat islets of Langerhans and HIT‐T15 cells causes significant DNA damage as measured by the ‘comet’ assay

et al. 1993

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We have used the comet assay (single cell gel electrophoresis) to measure nitric oxide-induced DNA damage in rat islets of Langerhans and insulin-containing HIT-T15 cells. Damage was induced following treatment with the nitric oxide donor SIN-l, which also releases superoxide, but was not reduced by exogenous superoxide dismutase, suggesting that nitric oxide itself, rather than superoxide or peroxynitrite may be the active species. The DNA damaging effect of nitric oxide was easily detectable at the earliest time point tested (15 min). Damage also resulted following induction of nitric oxide synthase by the cytokine interleukin-I/? in both islets and HIT-T15 cells and was prevented by replacing the substrate, arginine, with nitromonomethyl arginine. Thus intracellular levels of nitric oxide generated by interleukin-l/l-induced nitric oxide synthase were sufficient to cause DNA damage in islet cells and HIT-T15 cells.

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Section: Discussionmentioning

confidence: 99%

Endogenous nitric oxide induced by interleukin‐1β in rat islets of Langerhans and HIT‐T15 cells causes significant DNA damage as measured by the ‘comet’ assay

et al. 1993

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“…When DNA is exposed to UV light, in the presence of H 2 O 2 , this situation is leading to produce free hydroxyl radicals, and change native formation of DNA (scDNA), produced ocDNA and linear DNA (linDNA). [22][23][24][25][26][27] The addition of the water and ethanol extracts of HP at 25µg/ml-200µg/ml concentration to the reaction mixture prevent to change the formation of linDNA, and help to protect the native formation of DNA ( Figure.1). Figure 1 shows the electrophoretic band pattern of pBR322 plasmid DNA after exposed to UV/H 2 O 2 in the absence and presence of the extracts of HP in a dose dependent manner (concentration of the extracts ranging from 25µg/ml to 200µg/ml): first four lines are control samples, UV irritation of DNA in the absence of …”

Section: Antioxidant Activities Of Hpmentioning

confidence: 99%

“…The DNA protective activity of the water and ethanol extracts from stem, leaves and flowers of HP was analyzed using pBR322 plasmid DNA (Vivantis) in the presence of UV/H 2 O 2 described previously 21,22 with some modifications. For analysis, different experimental groups were used, including control (untreated pBR322 plasmid DNA, treated with H 2 O 2 and UV pBR322 plasmid DNA, treated with only H 2 O 2 pBR322 plasmid DNA and treated with only UV pBR322 plasmid DNA) and treated groups with different concentration of extracts (ranging from 25µg/ml to 200 µg/ml) and H 2 O 2 and UV.…”

Section: Dna Protective Potentialmentioning

confidence: 99%

Variation in Total Polyphenolic Contents, DNA Protective Potential and Antioxidant Capacity from Aqueous and Ethanol Extracts in Different Plant Parts of Hypericum perforatum L.

Şekeroğlu¹,

Urlu²,

Kulak³

et al. 2017

IJPER

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Background and Purpose: Hypericum perforatum belonging to the family Hypericaceae is a reputed medicinal plant including a wide ranges of important phytochemical components. Chlorogenic acid, rutin, hyperoside, quercitrin, quercetin, pseudohypericin, hypericin and hyperforin are of the major components. Crude extract and individual compounds of H. perforatum have been reported to exert antidepressant, antibiotic, and antitumoral activities. It is worthy to note that the quantity and efficacies of the crude extracts or individual compound are not constant, which are strongly influenced by different climatic conditions, harvesting times, harvested plant organs and post-harvest practices. Hence, numerous studies on H. perforatum collected from different parts of the World are carried out for their desired quality and biological efficacy. Methods: Wild collected plant materials were dried and preserved with a voucher specimen number and were extracted using maceration at room temperature for 24 h in dark. Subsequently, extracts were screened for their phenolic and flavonoid contents, plausible antioxidant activities using two methods namely DPPH radical scavenging and ferric-reducing antioxidant power (FRAP) assays and DNA protective activities. Results: The highlights of the study were are listed as 1) the highest total phenolic content in ethanol extracts of leaf, ii) the highest total flavonoid content in flower, iii) DPPH scavenging activity in leaf (80.51 %), flower (63.42 %) and stem (48.20 %), iv) highest ferric reduction capacity in ethanol extracts of stem were determined. Also, potent DNA protection activity was observed even at the lowest concentration value (25µg/ml) of the extracts. Conclusion: The phenolic content and strong antioxidant activities of ethanol extracts of different parts of the plant are reported. All the extracts exhibited strong DNA protective activities in response to the UV radiation in the presence of hydrogen peroxide.

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“…The comet assay was carried out according to [1]. In brief, 100 l of cell suspension was mixed with 100 l of 2% low-melting temperature agarose (Sigma) and spread on a slide pre-coated with 500 l of 0.5 % agarose.…”

Section: Comet Assaymentioning

confidence: 99%

“…The single-cell gel electrophoresis (SCGE), also known as the comet assay because damaged cells form a comet-shaped pattern after electrophoresis, is a sensitive method to measure genotoxicity and cytotoxicity of chemical and physical agents [1][2][3]. The comet assay has also been used to analyse the capacity of cellular DNA repair [4][5][6].…”

Section: Introductionmentioning

confidence: 99%

A cross-platform public domain PC image-analysis program for the comet assay

Końca

Lankoff

Banasik

et al. 2003

Mutation Research/Genetic Toxicology and Environmental Mutagene

654

289

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A microgel electrophoresis technique for the direct quantitation of DNA damage and repair in individual fibroblasts cultured on microscope slides

Cited by 233 publications

References 11 publications

Endogenous nitric oxide induced by interleukin‐1β in rat islets of Langerhans and HIT‐T15 cells causes significant DNA damage as measured by the ‘comet’ assay

Endogenous nitric oxide induced by interleukin‐1β in rat islets of Langerhans and HIT‐T15 cells causes significant DNA damage as measured by the ‘comet’ assay

Variation in Total Polyphenolic Contents, DNA Protective Potential and Antioxidant Capacity from Aqueous and Ethanol Extracts in Different Plant Parts of Hypericum perforatum L.

A cross-platform public domain PC image-analysis program for the comet assay

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