Tail Moment versus Tail Length—Application of an In Vitro Version of the Comet Assay in Biomonitoring for Genotoxicity in Native Surface Waters Using Primary Hepatocytes and Gill Cells from Zebrafish (Danio rerio)

2001 ◽  
Vol 49 (2) ◽  
pp. 187-196 ◽  
Author(s):  
A. Schnurstein ◽  
T. Braunbeck
Keyword(s):  
Comet Assay ◽  
Surface Waters ◽  
Danio Rerio ◽  
Tail Length ◽  
Tail Moment ◽  
Primary Hepatocytes ◽  
Gill Cells ◽  
Native Surface

In vitro toxicological assessment of flumethrin’s effects on MCF-7 breast cancer cells

2021 ◽  
pp. 096032712110227
Author(s):  
S Kara-Ertekin ◽  
S Yazar ◽  
M Erkan
Keyword(s):  
Breast Cancer ◽  
Cancer Cell Line ◽  
Tail Length ◽  
Tail Moment ◽  
High Concentration ◽  
Toxicological Assessment ◽  
Estrogenic Effects ◽  
Pyrethroid Pesticides ◽  
Mcf 7

Pyrethroid pesticides are frequently used for household insect control of insects and in agriculture and livestock. Flumethrin is a pyrethroid that is used against ectoparasites in many animals. The goal of this study was to evaluate the cytotoxic, apoptotic, genotoxic, and estrogenic effects of flumethrin on the mammalian breast cancer cell line (MCF-7). Compared with control groups, a dose-dependent decrease was observed in cell viability at concentrations of 100 µM and higher. The cytotoxic and apoptotic effects detected by LDH assay and AO/EtBr staining increased significantly at a concentration of 1000 µM. The expression of BCL2, which is an anti-apoptotic gene, significantly decreased, whereas BAX, TP53, and P21 expression significantly increased. The results of a comet assay indicated that flumethrin significantly changed tail length, tail % DNA, tail moment, and Olive tail moment in concentrations above 1 and 10 µM. In addition, a 0.1 µM concentration of flumethrin affected ERα receptor mediated cell proliferation and increased transcription of estrogen-responsive pS2 (TFF1) and progesterone receptor (PGR) genes. As a result, flumethrin-induced apoptosis and cytotoxicity at a high concentration, while induced genotoxicity even at lower concentrations. Flumethrin is an endocrine disrupting insecticide with estrogenic effects at very low concentrations.

A characterization of the ZFL cell line and primary hepatocytes as in vitro liver cell models for the zebrafish (Danio rerio)

2014 ◽  
Vol 147 ◽  
pp. 7-17 ◽  
Author(s):  
Marta Eide ◽  
Marte Rusten ◽  
Rune Male ◽  
Knut Helge Midtbø Jensen ◽  
Anders Goksøyr
Keyword(s):  
Cell Line ◽  
Danio Rerio ◽  
Liver Cell ◽  
Cell Models ◽  
Primary Hepatocytes

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

2009 ◽  
Vol 60 (1) ◽  
pp. 27-34 ◽  
Author(s):  
Verica Garaj-Vrhovac ◽  
Goran Gajski
Keyword(s):  
Comet Assay ◽  
Micronucleus Test ◽  
Human Lymphocytes ◽  
Tail Length ◽  
Bee Venom ◽  
Time Dependent ◽  
High Dose ◽  
Dependent Manner ◽  
High Concentration

Evaluation of the Cytogenetic Status of Human Lymphocytes After Exposure to a High Concentration of Bee Venom In VitroSeveral studies have reported radioprotective, antimutagenic, anti-inflammatory, 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 confirmed that the venom damaged the DNA molecule. Tail length, tail intensity, tail moment showed a significant increase (P<0.05). The percentage of long-tailed nuclei (LTN) with the tail length exceeding the 95th percentile also increased in a time-dependent manner. The micronucleus parameters (number of micronuclei, nucleoplasmic bridges, and nuclear buds) also showed a significant 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 find application in medicine.

scholarly journals Effect of Imidocarb Application on Oxidative DNA Damage Caused by Anaplasmosis

2021 ◽  
Vol 9 (12) ◽  
pp. 2308-2311
Author(s):  
Ahmet Cihat Öner ◽  
Adnan Ayan
Keyword(s):  
Dna Damage ◽  
Comet Assay ◽  
Dna Fragmentation ◽  
Oxidative Dna Damage ◽  
Clinical Signs ◽  
Tail Length ◽  
Assay Method ◽  
Control Group ◽  
Tail Moment ◽  
Before And After

This study was aimed to evaluate DNA fragmentation by using Comet assay in naturally infected sheep with Anaplasmosis before and after treatment with the Comet method, which shows DNA damage specifically. In the study, blood samples were collected from 10 Anaplosmosis infected and 10 healthy sheep. The anaplosmosis was diagnosed by clinical signs and symptoms. The infection was confirmed by Giemsa staining. The blood was collected from control group and infected group before and after the treatment, from the vena jugularis with the appropriate method. The DNA fragmentation was checked by using the Comet assay of blood cells. The data were analysed throught ANNOVA one-way. The result showed higher DNA fragmentation in sick animals diagnosed with anaplasmosis; tail length and tail moment values were found to be statistically significantly higher than the control group. When the data obtained after imidocarb (IMD) application were compared with obtained during the disease, a decreased DNA damage and tail moment was determined, however, these values higher than control. In this study, DNA damage and the extent of this damage were investigated by the Comet assay method using a healthy control group before and after treatment in animals with Anaplasmosis. When the findings obtained from the study were evaluated, it was seen that Anaplasma agents caused DNA damage and with the imidocarb application given for treatment, DNA damage was reduced and results close to healthy individuals were obtained.

scholarly journals Assessment of Cytogenetic Damage and Cholinesterases’ Activity in Workers Occupationally Exposed to Pesticides in Zamora-Jacona, Michoacan, Mexico

2021 ◽  
Vol 18 (12) ◽  
pp. 6269
Author(s):  
Rafael Valencia-Quintana ◽  
Rosa María López-Durán ◽  
Mirta Milić ◽  
Stefano Bonassi ◽  
Ma. Antonieta Ochoa-Ocaña ◽  
...  
Keyword(s):  
Dna Damage ◽  
Comet Assay ◽  
Whole Blood ◽  
Pesticide Exposure ◽  
Smoking Habit ◽  
Tail Length ◽  
Exposure Period ◽  
Control Group ◽  
Tail Moment ◽  
Cholinesterase Activities

Pesticides have been considered as potential chemical mutagens; however, little is known about toxic and genotoxic effects during pesticide application in Zamora-Jacona, Michoacan State in Mexico. This study sought to determine DNA damage and cholinesterase activities inhibitions in 54 agricultural workers exposed to complex mixtures of pesticides vs. control group (26 individuals) using Comet assay in peripheral whole blood, micronucleus (MN) test in oral mucosa cells, Cytokinesis-blocked MN assay in lymphocytes (L-CBMNcyt) and measuring AChE and BChE activities in whole blood and plasma samples, respectively. Exposed subjects demonstrated significantly elevated levels of primary (Comet assay: tail intensity, tail length, tail moment, Olive tail moment) and permanent DNA damage (MN assay: in blood/buccal cells; frequencies of nuclear buds, binucleated cells, cells with condensed chromatin, karyorrhexis, pyknosis, and karyolysis). However, inhibition of cholinesterase activities (AChE and BChE) was not observed in the workers. Confounding factors including sex, age, BMI, working exposure period, protection level, smoking habit (cigarettes per day units), alcohol consumption (weekly), medication, were considered in the analysis. These combined techniques demonstrated usefulness in the health hazards risks pesticide exposure assessment and suggested the need for periodic monitoring together with the education and the training of occupational workers for the safe application of potentially harmful pesticides.

Assessment of Antioxidant Effect of Beta-Glucan on the Whole Blood Oxidative DNA Damage with the Comet Assay in Colorectal Cancer

2021 ◽  
Vol 14 ◽  
Author(s):  
Necla Benlier ◽  
Nilay Uçar ◽  
Eda Öğüt ◽  
Havva Yeşil Çinkir ◽  
Mustafa Yildirim ◽  
...  
Keyword(s):  
Colorectal Cancer ◽  
Dna Damage ◽  
Comet Assay ◽  
Whole Blood ◽  
Oxidative Dna Damage ◽  
Tail Length ◽  
Antioxidant Effect ◽  
Control Group ◽  
Tail Moment ◽  
Beta Glucan

Objective: The present study aims to evaluate the antioxidant effect of beta glucan on oxidative DNA damage by comet assay. Methods: A total of 19 adult females and males diagnosed with stage 3-4 colorectal cancer and a control group of 20 age-matched healthy subjects were enrolled in the study. Blood samples of the participants were analyzed using Comet Assay for the parameters of DNA damage. Results: Significantly increased DNA damage was observed in patients versus control group as indicated by greater values of tail moment, tail percent DNA and tail length. Following incubation with β-glucan, a substantial reduction was found in the aforementioned parameters of DNA damage. Comet assay revealed significant levels of endogenous DNA damage in patients as shown by remarkable increases in the tail moment, the percentage of DNA in the tail and the tail length values, in comparison with the control group. Following treatment of fresh whole blood with β-glucan incubation, DNA damages were significantly reduced but lower values were observed after β-glucan incubation in the patient group versus control group. Conclusion: β-Glucan was found to reduce DNA damage substantially in colorectal cancer patients and show antimutagenic effects. Our results suggested that dietary β-glucan intake might be important in the genesis of colorectal cancer tumors.

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.

scholarly journals Investigating Genetic Damage in Peripheral Lymphocytes of Radiation Workers at Al-Tuwaitha Site in Iraq Using Four Genetic End-Points

2014 ◽  
Vol 8 (2) ◽  
pp. 5-10
Author(s):  
Amel J. Mutter ◽  
Abdulsahib K. Ali ◽  
Abdullah A. K. ◽  
Haider Y. L. ◽  
Ali H. F
Keyword(s):  
Comet Assay ◽  
Genetic Parameter ◽  
Tail Length ◽  
Control Group ◽  
Tail Moment ◽  
Genetic Damage ◽  
Peripheral Lymphocytes ◽  
Significant Difference ◽  
Radiation Workers ◽  
The Mean

The present study aims to use the biological techniques in a genotoxicity assessment of DNA damage in peripheral lymphocytes of radiation workers at Al-Tuwaitha site due to decommissioning to radioactive contamination as a result of work during January 2010 to December 2011. The subjects were divided into two groups: (i) 85 workers from radiation workers at Al-Tuwaitha site; (ii) 50 controls were matched non-smoking and no alcohol drink. Fresh blood samples were collected from the workers and controls. Four genetic parameter were studied using the micronucleus (MN) test, nuclear division index (NDI) test, the comet assay and hypoxanthine guanine phosphoribosyl transferase (HPRT) mutation assay. The results of the MN test showed that the average of MN per cell (Mean ± SE) in workers were 0.025 ± 0.0016 MN/ cells, which were significantly higher than those 0.010 ± 0.0006 MN/ cells in controls P< 0.01. While, the results of NDI test the average of NDI (Mean ± SE) in workers were 1.154 ± 0.0089 when compared with the control 1.322 ± 0.0117, which were significant increase p<0.01. It was found in the comet assay that the mean tail length (Mean ± SE) of radiation workers and controls were 17.69 ± 0.23 µm and 14.05 ± 0.13 µm, respectively. There was a significant difference between radiation workers and controls for mean tail length P < 0.01, but the difference between the mean tail moment (Mean ± SE) 14.22 ±0.21 of workers and mean tail moment 12.96± 0.15 of controls was not significant P> 0.01. Mean while, the results of the average of mutation frequency for HPRT were no significant differences rate for radiation workers compared with the control group P> 0.01. In conclusion, the results of our experiment suggest that the accumulation of genetic damage is detectable in peripheral lymphocytes of radiation workers at Al-Tuwaitha site. Also, the current results of frequency MN and NDI within of normal values according of the technical report of International Atomic Energy Agency (IAEA) No. 405, 2001.

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