scholarly journals Evaluation of DNA Damage in Common Carp (Cyprinus carpioL.) by Comet Assay for Determination of Possible Pollution in Lake Mogan (Ankara)

2011 ◽  
Vol 11 ◽  
pp. 1455-1461 ◽  
Author(s):  
İsmet Çok ◽  
Onur Kenan Ulutas ◽  
Öncü Okusluk ◽  
Emre Durmaz ◽  
Nilsun Demir
Keyword(s):  
Dna Damage ◽  
Comet Assay ◽  
Tail Length ◽  
Aquatic Organisms ◽  
Sewer System ◽  
Strand Breaks ◽  
Common Causes ◽  
The Common ◽  
Dna Strand

Contamination of the aquatic environment with various concentrations of pollutants results in unexpected threats to humans and wildlife. The consequences of exposure and metabolism of pollutants/xenobiotics, especially carcinogens and mutagens, can be suitably assessed by investigating severe events, such as DNA damage; for example, DNA adducts and DNA strand breaks. One of the commonly used techniques to detect DNA damage in aquatic organisms is single-cell gel electrophoresis (comet assay). This study was carried out usingCyprinus carpioin order to identify the possible pollution in Lake Mogan, near Ankara, Turkey, where the city's sewer system and pesticides used in agriculture are believed to be the common causes of pollution. From the comet assay, the tail length (μm), tail intensity (%), and tail moment values of fish caught from Lake Mogan were found to be 31.10 ± 10.39, 7.77 ± 4.51, 1.50 ± 1.48, respectively, whereas for clean reference sites they were found to be 22.80 ± 1.08, 3.47 ± 1.59, 0.40 ± 0.51, respectively. The values are statistically different from each other (p< 0.0001,p< 0.0001, andp< 0.0013, respectively). These results indicate that Lake Mogan may be polluted with substances that have genotoxic effects and constitute an early warning for the lake system. Further detailed research is needed to establish the source of the pollution and the chemicals responsible.

scholarly journals Assessment of genotoxicity and depuration of anthracene in the juvenile coastal fish Trachinotus carolinus using the comet assay

2013 ◽  
Vol 61 (4) ◽  
pp. 215-222 ◽  
Author(s):  
Fabio Matsu Hasue ◽  
Maria José de Arruda Campos Rocha Passos ◽  
Thaís da Cruz Alves dos Santos ◽  
Arthur José da Silva Rocha ◽  
Caroline Patrício Vignardi ◽  
...  
Keyword(s):  
Dna Damage ◽  
Comet Assay ◽  
Aquatic Organisms ◽  
Dna Strand Breaks ◽  
Time Dependent ◽  
Clean Water ◽  
Oxygen Species ◽  
Coastal Fish ◽  
Strand Breaks ◽  
Dna Strand

In the environment, anthracene is characterized as being persistent, bioaccumulative and toxic to aquatic organisms. Biotransformation of xenobiotic substances, such as anthracene, produces reactive oxygen species that may induce DNA strand breaks. The aim of the present study was to evaluate the DNA damage in juvenile T. carolinus exposed to different concentrations (8, 16 and 32 µg.L-1) of anthracene for 24 h in the dark then subsequently allowed to depurate in clean water for different periods of time (48, 96 or 144 h) using the comet assay. Our results show that anthracene is genotoxic to T. carolinus and that DNA damage was dose- and depuration/time- dependent. Anthracenegenotoxicity was observed in all experimental concentrations. Depuration seemed to be more efficient in fish exposed to thelowest anthracene concentration and maintained in clean water for 96 h.

DNA strand breaks and poly (ADP-ribose) polymerase activation induced by crystalline nickel subsulfide in MRC-5 lung fibroblast cells

1996 ◽  
Vol 15 (11) ◽  
pp. 891-897 ◽  
Author(s):  
ZX Zhuang ◽  
Y. Shen ◽  
HM Shen ◽  
V. Ng ◽  
CN Ong
Keyword(s):  
Dna Damage ◽  
Comet Assay ◽  
Tail Length ◽  
Dna Strand Breaks ◽  
Lung Fibroblast ◽  
Strand Breaks ◽  
Nickel Compounds ◽  
Dna Strand ◽  
Nickel Subsulfide

Nickel compounds are potent carcinogens. Their carcino genicity is believed to be associated with their solubility and cellular uptake. In the present study, we assessed the in vitro genotoxic effect of a water-insoluble nickel compound, crystalline nickel subsulfide (α-Ni 3S2) on human embryo lung fibroblast cell line (MRC-5 cells). DNA strand breaks was evaluated using single cell gel electrophoresis, or comet assay. The α-Ni3S2 induction of poly (ADP-ribose) polymerase (PADPRP), a nuclear enzyme associated with DNA damage and repair was also studied. Hydrogen peroxide (H2O2) was used as a reference compound. A dose-response relationship was found between α-Ni 3S2 concentrations (2.5 μg/cm2 to 20 μg/cm 2) and the comet tail length. The increase of PADPRP activity of α-Ni 3S2 treated MRC-5 cells was also significant and dose-dependent within the concentration range of 2.5 μg/ cm2 to 10 μg/cm 2. Close associations have been found between the comet length and PADPRP level for H2O2 (r=0.98) and α-Ni3S 2 (r=0.97). These results clearly suggest that α-Ni3S 2 is a potent agent in inducing DNA strand breaks, which may be closely related to its carcinogenic effects. Data from the present study also suggest that both comet assay and PADPRP determination are sensitive techniques for quantitative evaluation of DNA damage induced by nickel compounds.

Detection of DNA strand breaks by comet assay in sputum leucocytes of bitumen-exposed workers: A pilot study

2010 ◽  
Vol 29 (9) ◽  
pp. 721-729 ◽  
Author(s):  
B. Marczynski ◽  
M. Raulf-Heimsoth ◽  
B. Pesch ◽  
B. Kendzia ◽  
HU Käfferlein ◽  
...  
Keyword(s):  
Dna Damage ◽  
Comet Assay ◽  
Rank Correlation ◽  
Dna Strand Breaks ◽  
Strand Breaks ◽  
Blood Lymphocytes ◽  
Exposed Workers ◽  
Before And After ◽  
Dna Strand ◽  
Post Shift

DNA strand breaks were determined in leucocytes of induced sputum (IS) and compared with DNA strand breaks in blood lymphocytes from 42 bitumen-exposed workers pre and post shift. Comet assay results were expressed in arbitrary units based on visual scoring (sputum leucocytes) and Olive tail moment (OTM, blood lymphocytes). DNA damage in IS leucocytes was overall high but did not change during shift. Level of DNA strand breaks in IS samples correlated with total cell count and neutrophil content (Spearman rank correlation coefficient rs = 0.47, p = 0.001, rs= 0.48, p = 0.001, respectively) and with IL-8 concentration before and after shift (rs = 0.31, P = 0.048, and rs = 0.43, P = 0.005). DNA damage in IS was not associated with DNA strand breaks in blood lymphocytes (rs = —0.04, p = 0.802 before shift, rs = 0.27, p = 0.088 after shift). A higher level of DNA strand breaks was measured in blood lymphocytes before shift (median OTM 1.7 before and 1.3 after shift, p = 0.023). A strong correlation was found between the number of neutrophils and IL-8 concentration in IS before and after shift (rs = 0.77 and rs= 0.75, p < 0.001). This study showed an association between genotoxic and inflammatory effects in the lower airways and compared simultaneously DNA strand breaks in IS and blood of bitumen-exposed workers.

Use of comet assay for the risk assessment of oil- and chemical-industry workers

2014 ◽  
Vol 155 (47) ◽  
pp. 1872-1875 ◽  
Author(s):  
János Megyesi ◽  
Anna Biró ◽  
László Wigmond ◽  
Jenő Major ◽  
Anna Tompa
Keyword(s):  
Dna Damage ◽  
Occupational Exposure ◽  
Comet Assay ◽  
Oxidative Dna Damage ◽  
Microscopic Method ◽  
Monitoring Method ◽  
Strand Breaks ◽  
Cell Counts ◽  
Polycyclic Aromatic ◽  
Dna Strand

Introduction: The comet assay is a fluorescent microscopic method that is able to detect DNA strand-breaks even in non-proliferative cells in samples with low cell counts. Aim: The aim of the authors was to measure genotoxic DNA damage and assess oxidative DNA damage caused by occupational exposure in groups exposed to benzene, polycyclic aromatic carbohydrates and styrene at the workplace in order to clarify whether the comet assay can be used as an effect marker tool in genotoxicology monitoring. Method: In addition to the basic steps of the comet assay, one sample was treated with formamido-pirimidine-DNA-glycolase restriction-enzyme that measures oxidative DNA damage. Results: An increase was observed in tail moments in each group of untreated and Fpg-treated samples compared to the control. Conclusions: It can be concluded that occupational exposure can be detected with the method. The comet assay may prove to be an excellent effect marker and a supplementary technique for monitoring the presence or absence of genotoxic effects. Orv. Hetil., 2014, 155(47), 1872–1875.

scholarly journals DNA damage in human whole blood caused by radiopharmaceuticals evaluated by the comet assay

Mutagenesis ◽  
2019 ◽  
Vol 34 (3) ◽  
pp. 239-244 ◽  
Author(s):  
Heinz H Schmeiser ◽  
Karl-Rudolf Muehlbauer ◽  
Walter Mier ◽  
Ann-Christin Baranski ◽  
Oliver Neels ◽  
...  
Keyword(s):  
Dna Damage ◽  
Comet Assay ◽  
Dna Strand Breaks ◽  
Strand Breaks ◽  
Human Whole Blood ◽  
Time Points ◽  
Dna Migration ◽  
Activity Concentrations ◽  
Positron Emitters ◽  
Dna Strand

Abstract Radiopharmaceuticals used for diagnosis or therapy induce DNA strand breaks, which may be detectable by single-cell gel electrophoresis (called comet assay). Blood was taken from patients before and at different time points after treatment with radiopharmaceuticals; blood cells were investigated by the comet assay using the percentage of DNA in the tail as the critical parameter. Whereas [225Ac]Ac-prostate-specific membrane antigen (PSMA)-617 alpha therapy showed no difference relative to the blood sample taken before treatment, beta therapy with [177Lu]Lu-PSMA-617 3 h post-injection revealed a small but significant increase in DNA strand breaks. In blood of patients who underwent positron emission tomography (PET) with either [18F]2-fluor-2-deoxy-D-glucose (FDG) or [68Ga]Ga-PSMA-11, an increase of DNA migration determined by the comet assay was not found when analysed at different time points (2–70 min) after intravenous tracer injection. Human whole blood was incubated with the targeted clinically relevant therapeutic radiopharmaceuticals [225Ac]Ac-PSMA-617, [177Lu]Lu-PSMA-617 and [90Y]Y-DOTA(0)-Phe(1)-Tyr(3)-octreotide (DOTA-TOC) at different activity concentrations (kBq/ml) for 5 days and then analysed by the comet assay. DNA damage increased with higher concentrations of all radiolabeled compounds tested. [177Lu]Lu-PSMA-617 caused higher blood cell radiotoxicity than equal activity concentrations of [90Y]Y-DOTA-TOC. Likewise, whole human blood was exposed to the positron emitters [18F]FDG and [68Ga]Ga-PSMA-11 in vitro for 24 h with activity concentrations ranging between 5 and 40 MBq/ml. The same activity concentration dependent elevated DNA migration was observed for both compounds although decay energies are different. This study demonstrated that the amount of DNA damage detected by the comet assay in whole human blood is similar among different positron emitters and divergent by a factor of 200 between alpha particles and beta radiation.

scholarly journals Single-Dose Toxicity of Individual and Combined Sterigmatocystin and 5-Methoxysterigmatocistin in Rat Lungs

Toxins ◽  
2020 ◽  
Vol 12 (11) ◽  
pp. 734
Author(s):  
Daniela Jakšić ◽  
Ida Ćurtović ◽  
Domagoj Kifer ◽  
Dubravka Rašić ◽  
Nevenka Kopjar ◽  
...  
Keyword(s):  
Dna Damage ◽  
Comet Assay ◽  
Lavage Fluid ◽  
Lung Weight ◽  
Lactate Dehydrogenase Activity ◽  
Strand Breaks ◽  
Rat Lungs ◽  
Male Wistar Rats ◽  
Dna Strand ◽  
Factor Α

Sterigmatocystin (STC) and 5-methoxysterigmatocystin (5-M-STC) are mycotoxins produced by common damp indoor Aspergilli series Versicolores. Since both STC and 5-M-STC were found in the dust of indoor occupational and living areas, their occupants may be exposed to these mycotoxins, primarily by inhalation. Thus, STC and 5-M-STC were intratracheally instilled in male Wistar rats using doses (0.3 mg STC/kg of lung weight (l.w.); 3.6 mg 5-M-STC/kg l.w.; toxin combination 0.3 + 3.6 mg/kg l.w.) that corresponded to concentrations detected in the dust of damp indoor areas in order to explore cytotoxicity, vascular permeability, immunomodulation and genotoxicity. Single mycotoxins and their combinations insignificantly altered lactate-dehydrogenase activity, albumin, interleukin-6, tumor necrosis factor-α and chemokine macrophage inflammatory protein-1α concentrations, as measured by ELISA in bronchioalveolar lavage fluid upon 24 h of treatment. In an alkaline comet assay, both mycotoxins provoked a similar intensity of DNA damage in rat lungs, while in a neutral comet assay, only 5-M-STC evoked significant DNA damage. Hence, naturally occurring concentrations of individual STC may induce DNA damage in rat lungs, in which single DNA strand breaks prevail, while 5-M-STC was more responsible for double-strand breaks. In both versions of the comet assay treatment with STC + 5-M-STC, less DNA damage intensity occurred compared to single mycotoxin treatment, suggesting an antagonistic genotoxic action.

Detection of Oxidised Purines and UV-induced Photoproducts in DNA of Single Cells, by Inclusion of Lesion-specific Enzymes in the Comet Assay

1996 ◽  
Vol 24 (3) ◽  
pp. 405-411 ◽  
Author(s):  
Mária Dušinská ◽  
Andrew Collins
Keyword(s):  
Dna Damage ◽  
Comet Assay ◽  
Single Cells ◽  
Dna Strand Breaks ◽  
Human Populations ◽  
Dna Damage And Repair ◽  
Strand Breaks ◽  
Dna Lesion ◽  
Dna Strand ◽  
Uv Photoproducts

The comet assay (single cell gel electrophoresis) is a rapid, very sensitive method for the detection of DNA strand breaks at the level of single cells, which is now being applied in genotojricity testing. We modified this method for the detection of a variety of kinds of DNA lesion, by treating nucleoid DNA in the gel with either formamidopyrimidine-DNA glycosylase (which recognises ring opened purines, 8-hydroxyguanine and apurinic/apyrimidinic sites), or uvrABC excinuclease (uvrABC; which has a rather broad specificity, including bulky lesions and UV photoproducts). By using this modified assay, we demonstrate the removal of DNA strand breaks and oxidised purines upon incubating cells after treatment with hydrogen peroxide. This modification clearly increases the usefulness of the assay for the analysis of DNA damage and repair, for screening human populations for DNA damage, and for testing novel chemicals for genotoxicity.

scholarly journals Assessment of Cellular Damage by Comet Assay After Photodynamic Therapy in vitro

2004 ◽  
Vol 47 (4) ◽  
pp. 327-329 ◽  
Author(s):  
Jaroslav Maceček ◽  
Hana Kolářová ◽  
Jitka Psotová ◽  
Robert Bajgar ◽  
Martin Huf ◽  
...  
Keyword(s):  
Dna Damage ◽  
Photodynamic Therapy ◽  
Comet Assay ◽  
Single Cells ◽  
Cellular Damage ◽  
Cytotoxic Action ◽  
Dna Breaks ◽  
Strand Breaks ◽  
Dna Strand

The aim of this study was analysis of DNA damage in the cell line of the human melanoma G361 after photodynamic therapy (PDT) by comet assay. Photodynamic therapy is based on cytotoxic action of sensitizers (10 µM ZnTPPS4 fixed into 1 mM cyclodextrin hpβCD) and light with a suitable wavelength. Single-cell gel electrophoresis (SCGE, comet assay) is a rapid and sensitive method for detecting DNA strand breaks at the level of single cells. Great amount of DNA damage was detected with the dose of irradiation of 0.1; 0.5 J and 2.5 J.cm-2. Only radiation dose of visible light in the presence of sensitizers can induce DNA breaks of tumour cells. Cells with DNA damage appear as fluorescent comets with tails of DNA fragmentation. In contrast, cells with undamage DNA appear as round spots, because their intact DNA does not migrate out of the cell.

DNA damage in lymphocytes after irradiation with 211At and 188Re

2009 ◽  
Vol 48 (06) ◽  
pp. 221-226 ◽  
Author(s):  
M. Wendisch ◽  
G. Wunderlich ◽  
R. Freudenberg ◽  
J. Kotzerke ◽  
R. Runge
Keyword(s):  
Dna Damage ◽  
Comet Assay ◽  
Mononuclear Cells ◽  
Dna Lesions ◽  
Alpha Particles ◽  
Alkaline Comet Assay ◽  
Peripheral Blood Mononuclear ◽  
Strand Breaks ◽  
Neutral Comet Assay ◽  
Dna Strand

Summary Aim: Ionising radiation produces many types of DNA lesions of different complexity. High linear energy transfer (LET) types of radiation are biological more effective than low LET radiation. In the present work we applied the single cell gel electrophoreses (comet assay) to study the induction of initial DNA damage, efficiency of repair and residual DNA damage in lymphocytes after treatment with 211At and 188Re. Methods: Peripheral blood mononuclear cells (PBMC) were isolated from heparinized blood of healthy donors and irradiated with 211At and 188Re at different doses. The comet assay was performed under alkaline and neutral conditions in order to detect the initial DNA damage and its repair. The measure of damage was % tail DNA (percentage of DNA in the tail). Results: After treatment of cells with 188Re the initial DNA damage (% tail DNA) detected with the alkaline comet assay was higher than the damage measured for 211At. The neutral comet assay estimated higher tail intensities for 211At in contrast to 188Re. Compared with the complete repair (10%) after irradiation with 188Re, the radiotoxicity of alpha particles indicated reduced rejoining of DNA strand breaks (60–80% residual damage). Rejoining of DNA damage measured by the neutral comet method detected about 70% unrepaired strand breaks for 211At and 188Re. Conclusions: There are major differences between the repair of strand breaks caused by 188Re and 211At detected by the alkaline comet assay. The DNA-damage induced by the high LET Emitter 211At remains nearly unrepaired detected by both alkaline and neutral comet assay. Represented data following irradiation of lymphocytes with alpha and beta particles demonstrated higher biological effectiveness of 211At by factors of 2.0–2.5.

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