A modified alkaline Comet assay for in vivo detection of oxidative DNA damage in Drosophila melanogaster

Author(s):  
A.K. Shukla ◽  
P. Pragya ◽  
D. Kar Chowdhuri
Molecules ◽  
2019 ◽  
Vol 24 (8) ◽  
pp. 1560 ◽  
Author(s):  
Nevenka Kopjar ◽  
Nino Fuchs ◽  
Suzana Žunec ◽  
Anja Mikolić ◽  
Vedran Micek ◽  
...  

Currently we are faced with an ever-growing use of Δ9-tetrahydrocannabinol (THC) preparations, often used as supportive therapies for various malignancies and neurological disorders. As some of illegally distributed forms of such preparations, like cannabis oils and butane hash oil, might contain over 80% of THC, their consumers can become intoxicated or experience various detrimental effects. This fact motivated us for the assessments of THC toxicity in vivo on a Wistar rat model, at a daily oral dose of 7 mg/kg which is comparable to those found in illicit preparations. The main objective of the present study was to establish the magnitude and dynamics of DNA breakage associated with THC exposure in white blood and brain cells of treated rats using the alkaline comet assay. The extent of oxidative stress after acute 24 h exposure to THC was also determined as well as changes in activities of plasma and brain cholinesterases (ChE) in THC-treated and control rats. The DNA of brain cells was more prone to breakage after THC treatment compared to DNA in white blood cells. Even though DNA damage quantified by the alkaline comet assay is subject to repair, its elevated level detected in the brain cells of THC-treated rats was reason for concern. Since neurons do not proliferate, increased levels of DNA damage present threats to these cells in terms of both viability and genome stability, while inefficient DNA repair might lead to their progressive loss. The present study contributes to existing knowledge with evidence that acute exposure to a high THC dose led to low-level DNA damage in white blood cells and brain cells of rats and induced oxidative stress in brain, but did not disturb ChE activities.


2019 ◽  
Vol 12 (2) ◽  
pp. 163-172 ◽  
Author(s):  
D. Rašić ◽  
D. Želježić ◽  
N. Kopjar ◽  
D. Kifer ◽  
M. Šegvić Klarić ◽  
...  

The study aimed to check whether ochratoxin A (OTA) and citrinin (CIT) increase DNA damage in the kidney and liver of male Wistar rats (alkaline comet assay), clarify the oxidative nature of DNA damage (hOGG1-modified comet assay), and verify whether resveratrol (RSV) could ameliorate OTA+CIT-induced genotoxicity. Rats were treated orally with OTA (0.125 and 0.250 mg/kg bodyweight (bw)) and CIT (2 mg/kg bw), OTA+CIT combinations and OTA+CIT+RSV (0.250+2+20 mg/kg bw) for 21 days. Both alkaline and hOGG1-modified comet assay showed that DNA damage was more severe in rat kidneys than in liver following mycotoxin treatment. Alkaline comet assay revealed a higher intensity of DNA damage, particularly as measured by tail intensity in the kidneys. Both tail length and tail intensity were OTA dose-dependent, but in combined OTA+CIT treatment these values were similar to CIT alone and lower than in animals treated with single OTA, possibly due to induction of apoptosis. hOGG1-modified comet showed that OTA+CIT evoked greater oxidative DNA damage than single mycotoxins. RSV did not reduce DNA damage measured by alkaline comet assay, but hOGG1-modified comet showed that RSV ameliorated OTA+CIT genotoxicity in the kidneys. Apart from oxidative stress, other mechanisms of DNA damage are involved in OTA and CIT genotoxicity. In rat kidneys RSV can reduce but not overcome oxidative DNA damage induced by combined OTA and CIT.


Mutagenesis ◽  
2005 ◽  
Vol 20 (4) ◽  
pp. 285-290 ◽  
Author(s):  
Hifzur R. Siddique ◽  
D.Kar Chowdhuri ◽  
D.K. Saxena ◽  
Alok Dhawan

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