DNA damage assessment by comet assay of human lymphocytes exposed to jet propulsion fuels

2002 ◽  
Vol 40 (1) ◽  
pp. 18-23 ◽  
Author(s):  
Shawna M. Jackman ◽  
Geraldine M. Grant ◽  
Christopher J. Kolanko ◽  
David A. Stenger ◽  
Joginder Nath
Keyword(s):  
Dna Damage ◽  
Comet Assay ◽  
Damage Assessment ◽  
Human Lymphocytes ◽  
Jet Propulsion

Imatinib (STI571) Induces DNA Damage in BCR/ABL-Expressing Leukemic Cells but Not in Normal Lymphocytes.

Blood ◽  
2004 ◽  
Vol 104 (11) ◽  
pp. 4353-4353
Author(s):  
Janusz Blasiak ◽  
Jozef Drzewoski ◽  
Tomasz Poplawski ◽  
Agnieszka Czechowska
Keyword(s):  
Dna Damage ◽  
Comet Assay ◽  
Human Lymphocytes ◽  
Direct Interaction ◽  
K562 Cells ◽  
Dna Lesions ◽  
Dna Glycosylase ◽  
Leukemic Cells ◽  
Drug Induced ◽  
Strand Breaks

Abstract Imatinib (STI571) is a 2-phenylaminopyrimidine derivative used mostly in the treatment of chronic myeloid leukaemia. It targets specifically the BCR/ABL oncogenic tyrosine kinase, inhibiting its activity. Using the alkaline comet assay we showed that STI571 at concentrations ranging from 0.05 to 2 μM induced DNA damage in human leukemic K562 cells expressing the BCR/ABL oncogene, whereas it had no effect in normal human lymphocytes. Because the extent of DNA damage observed in the neutral and pH 12.1 versions of the comet assay was much lesser than in the alkaline version, we concluded that the drug induced DNA alkali-labile sites rather than strand breaks. Imatinib did not induce DNA strand breaks in the direct interaction with DNA as examined by the plasmid relaxation assay. K562 cells were unable to repair H2O2-induced DNA damage during a 120-min incubation, if they had been preincubated with STI571, whereas normal lymphocytes did so within 60 min. Pre-treatment of K562 cells with vitamins A, C and E reduced the extent of DNA damage evoked by STI571. Similar results brought experiments with the nitrone spin traps POBN and PBN, suggesting that free radicals may be involved in the formation of DNA lesions induced by STI571 in K562 cells. These cells exposed to imatinib and treated with endonuclease III, formamidopyrimidine-DNA glycosylase and 3-methyladenine-DNA glycosylase II, the enzymes recognizing oxidized and alkylated bases, displayed greater extent of DNA damage than those not treated with these enzymes.

scholarly journals Effects of photopolymerisation on genotoxicity of composite adhesives in the comet assay

Genetika ◽  
2016 ◽  
Vol 48 (2) ◽  
pp. 617-627
Author(s):  
Stefan Dacic ◽  
Ninoslav Djelic ◽  
Milena Radakovic ◽  
Nada Lakic ◽  
Aleksandar Veselinovic ◽  
...  
Keyword(s):  
Dna Damage ◽  
Comet Assay ◽  
Human Lymphocytes ◽  
Negative Control ◽  
In Vivo Studies ◽  
Halogen Lamp ◽  
Genotoxic Effects ◽  
Significant Difference ◽  
Isolated Human Lymphocytes

Certain in vivo studies have shown that the application of adhesives directly onto the open pulp or on a thin layer of dentin causes inflammation and pulpal abscesses. This reaction is related to toxic effects of monomers from adhesives. It has been confirmed that after proper illumination the adhesives become less toxic. The aim of the study was to examine genotoxicity of non-polymerised, partly polymerised and polymerised adhesives on isolated human lymphocytes using the alkaline Comet assay. Adper Single bond2 and Adper Easy One/3M ESPE adhesive photopolymerisation was performed by Elipar Highlight 3M ESPE halogen lamp for 0, 10 and 40 sec, at final concentrations of 100, 200, 500 and 1000 ?g/mL. With both adhesives, photopolymerisation at 0 and 10 seconds showed statistically significant increase in DNA damage in comparision to the negative control (solvent). On the other hand, after 40 seconds of photopolymerisation of both adhesives in all tested concentrations, the degree of DNA damage in Comet assay had no significant difference (P>0.05, ?2 test) compared to the negative control. Therefore, only the 40 seconds of photopolymerisation prevented genotoxic effects of both adhesives in the Comet assay.

scholarly journals EVALUATION OF CYTOTOXIC AND GENOTOXIC EFFECTS OF ZERUMBONE ON COLON ADENOCARCINOMA COLO205 CELLS AND HUMAN LYMPHOCYTES

2017 ◽  
Vol 9 (10) ◽  
pp. 92
Author(s):  
Rima Thiyam ◽  
Mangamoori Lakshmi Narasu
Keyword(s):  
Dna Damage ◽  
Cell Death ◽  
Comet Assay ◽  
Apoptotic Cell ◽  
Morphological Changes ◽  
Human Lymphocytes ◽  
Genotoxic Effect ◽  
Colorectal Cancer Cell Line ◽  
Dependent Manner ◽  
Ic50 Values

Objective: The objective of the present study was to investigate the growth inhibitory effect, apoptosis initiation and genotoxic effect of zerumbone (ZER), a phytochemical and cisplatin, a chemotherapeutic drug on human colorectal cancer cell line COLO205 and normal human lymphocytes.Methods: The antiproliferative activity of ZER and cisplatin (positive control) on COLO205 cells and lymphocytes was analysed by 3( 4, 5-dimethylthiazol-2-yl)-2-5-diphenyltetrazolium bromide (MTT) assay. Morphological analysis of the cells was studied by using inverted phase contrast microscope. Propidium iodide staining method was used to observe the apoptotic morphological changes in the treated cells. Finally comet assay was conducted to observe the extent of DNA damage induced by ZER and cisplatin on COLO205 and lymphocytes.Results: ZER and cisplatin exhibited growth inhibition in a dose and time dependent manner against COLO205 with no considerable effect on lymphocytes. The IC50 values of ZER on COLO205 for 24h, 48h and 72h were 19 µg/ml, 10 µg/ml and 5 µg/ml. Comparatively the IC50 values of cisplatin on COLO205 for 24h, 48h and 72h were 38 µg/ml, 24 µg/ml and 15 µg/ml.  Morphological changes such as cell shrinkage, membrane blebbing and nuclear condensation were observed in COLO205 while no significant change was observed in lymphocytes. Fluorescence imaging studies confirmed apoptotic cell death in treated COLO205 cells while no significant cell death was observed in treated lymphocytes. Comet assay revealed significant DNA damage in treated COLO205 cells.Conclusion: The present study demonstrated the cytotoxic and genotoxic effect of ZER and cisplatin on COLO205 cells. These drugs showed no significant effect on lymphocytes.

Chaga mushroom extract inhibits oxidative DNA damage in human lymphocytes as assessed by comet assay

BioFactors ◽  
2004 ◽  
Vol 21 (1-4) ◽  
pp. 109-112 ◽  
Author(s):  
Yoo Kyoung Park ◽  
Hyang Burm Lee ◽  
Eun-Jae Jeon ◽  
Hack Sung Jung ◽  
Myung-Hee Kang
Keyword(s):  
Dna Damage ◽  
Comet Assay ◽  
Oxidative Dna Damage ◽  
Human Lymphocytes ◽  
Mushroom Extract

scholarly journals Genotoxic effects of diazinon on human peripheral blood lymphocytes / Genotoksično djelovanje diazinona na limfocite ljudske periferne krvi

2015 ◽  
Vol 66 (2) ◽  
pp. 153-158 ◽  
Author(s):  
Fulya Dilek Gökalp Muranli ◽  
Martin Kanev ◽  
Kezban Ozdemir
Keyword(s):  
Dna Damage ◽  
Comet Assay ◽  
Peripheral Blood ◽  
Human Peripheral Blood ◽  
Human Lymphocytes ◽  
Peripheral Blood Lymphocytes ◽  
Blood Lymphocytes ◽  
Human Peripheral Blood Lymphocytes ◽  
Current Assessment ◽  
High Level

Abstract The aim of this study was to evaluate the genetic damage in human peripheral blood lymphocytes following 24 and 48- hour exposure to a commercial diazinon formulation Basudin 60EM® at concentrations between 0.01 and 40 μg mL-1. For this purpose we used the micronucleus (MN), fluorescence in situ hybridization (FISH), and alkaline single cell gel electrophoresis (comet) assay. Diazinon significantly increased the frequency of micronucleated cells compared to control. Forty-eight-hour exposure increased this frequency even at lower concentrations (0.01-10 μg mL-1). The FISH results revealed aneugenic effects at 10 μg mL-1. The comet assay also confirmed DNA damage at concentrations between 10 and 40 μg mL-1. Our findings have confirmed the genotoxic potential of diazinon and its cytotoxic effect on human lymphocytes. The increased DNA damage in our study raises concern about the current assessment of the health risk posed by this pesticide and calls for a high level of caution in agricultural and household use.

scholarly journals A Silver Staining Method for Single-cell Gel Assay

2001 ◽  
Vol 49 (9) ◽  
pp. 1183-1186 ◽  
Author(s):  
Silvina B. Nadin ◽  
Laura M. Vargas–Roig ◽  
Daniel R. Ciocca
Keyword(s):  
Dna Damage ◽  
Comet Assay ◽  
Single Cell ◽  
Silver Staining ◽  
Staining Method ◽  
Human Lymphocytes ◽  
Fluorescence Staining ◽  
High Quality ◽  
Dna Damage And Repair ◽  
Silver Staining Method

The single-cell gel assay (comet assay) is a very useful microelectrophoretic technique for evaluation of DNA damage and repair in individual cells. Usually, the comets are visualized and evaluated with fluorescent DNA stains. This staining requires specific equipment (e.g., a high-quality fluorescence microscope), the slides must be analyzed immediately, and they cannot be stored for long periods of time. Here we describe, using human lymphocytes, some modifications of the silver staining for comets that significantly increase the sensitivity/reproducibility of the assay. This silver staining was compared with fluorescence staining and commercial silver stains. (J Histochem Cytochem 49:1183–1186, 2001)

Measurement of Menadione-Mediated DNA Damage in Human Lymphocytes Using the Comet Assay

1997 ◽  
Vol 26 (2) ◽  
pp. 113-124 ◽  
Author(s):  
J. A. Woods ◽  
A. J. Young ◽  
I. T. Gilmore ◽  
A. Morris ◽  
R. F. Bilton
Keyword(s):  
Dna Damage ◽  
Comet Assay ◽  
Human Lymphocytes
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