Comet assay for nuclear DNA damage

Piperakis, Stylianos M.; Visvardis, Evangelos-E.; Tassiou, Aspasia

doi:10.1016/s0076-6879(99)00125-1

Cited by 72 publications

(44 citation statements)

References 24 publications

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“…There is a variety of possible modifi cations of the assay which facilitate the detection of single-strand DNA breaks, alkali-labile sites, double-strand DNA breaks, incomplete excision repair sites, and interstrand crosslinks (33). The comet assay can also be used to assess DNA fragmentation associated with cell death or apoptosis (34,35).…”

Section: Discussionmentioning

confidence: 99%

Evaluation of the Cytogenetic Status of Human Lymphocytes After Exposure to a High Concentration of Bee Venom In Vitro

Garaj-Vrhovac

Gajski

2009

Archives of Industrial Hygiene and Toxicology

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Several studies have reported radioprotective, antimutagenic, anti-infl ammatory, antinociceptive, and anticancer effects of bee venom both in the cell and the whole organism. The aim of this study was to assess the effects of a single high dose of 100 µg mL -1 of whole bee venom in human lymphocytes in vitro over a variety of time spans (from 10 min to 24 h). After the treatment, we used the comet assay and micronucleus test to see the effect of bee venom on the cell. The comet assay confi rmed that the venom damaged the DNA molecule. Tail length, tail intensity, tail moment showed a signifi cant increase (P<0.05). The percentage of long-tailed nuclei (LTN) with the tail length exceeding the 95 th percentile also increased in a time-dependent manner. The micronucleus parameters (number of micronuclei, nucleoplasmic bridges, and nuclear buds) also showed a signifi cant time-dependent increase (P<0.05). This research indicates that high concentrations of bee venom can lead to cellular instability. Further research is needed to understand the mechanism of action of bee venom and its components in human cells and to see if this natural product may fi nd application in medicine.

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Section: Discussionmentioning

confidence: 99%

Evaluation of the Cytogenetic Status of Human Lymphocytes After Exposure to a High Concentration of Bee Venom In Vitro

Garaj-Vrhovac

Gajski

2009

Archives of Industrial Hygiene and Toxicology

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“…Therefore, chemical alterations of nucleic acids, such as breaks of hydrogen bounds, breaks of base-sugar binding, sugar oxidation, break of nucleotide strand and release of terminal phosphates caused by reaction of free radicals can be sensitively detected by means of the technique used. 2,18) The exact mechanism of action in protecting mice from lethal effects of acute WBI by propolis and related flavonoids is not known. These results indicate that preventive treatment of irradiated mice with WSDP and related flavonoids significantly decreased the levels of primary DNA damage in their white blood cells and increased life span of mice, 19) as compared to untreated animals.…”

Section: Discussionmentioning

confidence: 99%

Evaluation of the Radioprotective Effects of Propolis and Flavonoids in Gamma-Irradiated Mice: The Alkaline Comet Assay Study

Benković

Oršolić

Knežević

et al. 2008

Biological & Pharmaceutical Bulletin

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Ionizing radiation is classified as a potent carcinogen. When interacting with living cells, it causes a variety of changes depending on exposed and absorbed dose, duration of exposure and interval after exposure, and susceptibility of tissues.1) It is well established that ionizing radiation by direct ionization and through generation of toxic free radicals causes single strand breaks, double strand breaks, oxidative damage to sugar and base residues, chromosomal aberration and mutation. 2)Since the radiation effects are not discriminate, transient and/or permanent injury to normal tissues and cells are unavoidable.3) Efforts to reduce toxicity to normal tissue cells and organs have led to the search for cytoprotective agents. Unfortunately, most of chemical radioprotectors (AET, WR 2721, WR 1065) have shown toxic side effects that limit their use in medical practice. 4) Investigations for effective and non toxic compounds with radioprotection capability led to increasing interest in naturally occurring antioxidants.Propolis is adhesive resinous substance manufactured by honey bees from leaf, bud and sap of trees and flower blossoms. Major constituents of propolis are flavonoids, organic acids, phenols, various kinds of enzymes, vitamins and minerals.5,6) Because of its broad spectrum of biological properties the interest in propolis as harmless medicine has been increased.7) Propolis and its active substances flavonoids showed antibacterial, 7) analgesic/anti-inflammatory, 7) antioxidant, 6) prooxidant, 8) immunoenchancement, 9) antiproliferative activity in cultured human tumor cells 10) and antitumor activity in mice.11) Antioxidant activity of flavonoids is based on ability of direct free radicals scavenging or stabilizing the reactive oxygen species (ROS) by interacting with the reactive compound of the radical. Because of the high reactivity of the hydroxyl substituents of the flavonoids, radicals are made inactive. 7) ROS in the presence of iron resulted in lipid peroxidation. Some flavonoids are known to chelate iron. 7)Flavonoids can also increase the function of the endogenous antioxidant enzyme systems; superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GPx), glutathione reductase (GR) and gluthation.12) Furthermore, antioxidant effects may be a result of a combination of radical scavenging and an interaction with enzyme functions.Genotoxicity of ionizing radiation was studied with different approaches using sensitive methods to measure a single and double strand breaks (SSB, DSB) of DNA; mostly used tests are comet assay for SSB and micronucleus test for DSB.The aim of this study was to evaluate radioprotective effects of propolis and single flavonoids, caffeic acid, chrysin and naringin in the whole-body irradiated (WBI) mice with two doses of g-irradiation using an alkaline comet assay. The efficiency of tested compounds was evaluated on the white blood cells of mice treated preventively and therapeutically. The possible genotoxic effects of all compounds were also assessed on non...

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“…Moreover, it enables the study of different DNA repair pathways such as base excision and nucleotide excision repair. In addition to the above, DNA fragmentation associated with cell death or related to apoptosis can be evaluated [10,[17][18][19].…”

Section: Introductionmentioning

confidence: 99%

Assessment of chemotherapy‐induced DNA damage in peripheral blood leukocytes of cancer patients using the alkaline comet assay

Kopjar

Garaj‐Vrhovac

Milas

2001

Teratog Carcinog Mutagen

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The alkaline comet assay was employed to assess the pre-and post-treatment levels of in vivo DNA damage in peripheral blood leukocytes of cancer patients. During the study all patients were given antineoplastic drugs, mainly as polychemotherapy. To quantify the DNA damage, two different comet parameters were evaluated: the tail length and the tail moment. Our results indicate marked interindividual variations between baseline DNA damage in peripheral blood leukocytes recorded among cancer patients prior to the chemotherapy. After intravenous administration of various antineoplastic drugs, a significantly increased level of DNA damage in all cancer patients compared to their pre-treatment values was recorded The highest level of DNA damage was seen following administration of 5-fluorouracil, adriamycin, and cisplatin (FAP protocol). The results indicate that administration of antineoplastic drugs in standard protocols is accompanied by significant DNA damage in peripheral blood leukocytes. In order to diminish the potential risks of developing second neoplasms, a continuous biomonitoring of cancer patients after the ending of chemotherapy becomes important. Despite their limitations, present results confirm the usefulness of the alkaline comet assay as a sensitive biomarker of exposure that enables rapid and simple detection of primary DNA damage in peripheral blood leukocytes of cancer patients. Together with standard cytogenetic endpoints, the comet assay provides a powerful technique for the routine detection of critical DNA lesions produced after administration of antineoplastic drugs in the clinical settings. Teratogenesis Carcinog. Mutagen. 22:13-30, 2002.

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Comet assay for nuclear DNA damage

Cited by 72 publications

References 24 publications

Evaluation of the Cytogenetic Status of Human Lymphocytes After Exposure to a High Concentration of Bee Venom In Vitro

Evaluation of the Cytogenetic Status of Human Lymphocytes After Exposure to a High Concentration of Bee Venom In Vitro

Evaluation of the Radioprotective Effects of Propolis and Flavonoids in Gamma-Irradiated Mice: The Alkaline Comet Assay Study

Assessment of chemotherapy‐induced DNA damage in peripheral blood leukocytes of cancer patients using the alkaline comet assay

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