Quantification of applied dose in irradiated citrus fruits by DNA Comet Assay together with image analysis

Çetinkaya, Nurcan; Erçin, Demet; Özvatan, Sümer; Erel, Yakup

doi:10.1016/j.foodchem.2015.07.027

Cited by 15 publications

(11 citation statements)

References 15 publications

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“…In addition, the in vivo studies also used cells obtained from a numerous different species including mainly rat, fish, mouse and worm. Some few studies also investigated DNA damage in bacteria (Danevcic et al 2016), plant cells (Cetinkaya et al 2016;Faisal et al 2016;Lanier et al 2015), amoebae (Kusrini et al 2016) or hydra (Zeeshan et al 2016). Whereas in the in vitro studies, a wide range of cell types belonging to different organ systems (e.g., A549, HCT 116, HEPG2, MCF-7, MRC-5, BEAS-2B, CHO-K1, and HEK293) were used, the in vivo studies were mainly based on cells from the blood system (about 61 %) like erythrocytes obtained from, e.g., fish or lymphocytes from, e.g., humans.…”

Section: Current Workmentioning

confidence: 99%

Comet assay: an essential tool in toxicological research

2016

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The comet assay is a versatile, reliable, cost-efficient, and fast technique for detecting DNA damage and repair in any tissue. It is useable in almost any cell type and applicable to both eukaryotic and prokaryotic organisms. Instead of highlighting one of the numerous specific aspects of the comet assay, the present review aims at giving an overview about the evolution of this widely applicable method from the first description by Ostling and Johanson to the OECD Guideline 489 for the in vivo mammalian comet assay. In addition, methodical aspects and the influence of critical steps of the assay as well as the evaluation of results and improvements of the method are reviewed. Methodical aspects regarding oxidative DNA damage and repair are also addressed. An overview about the most recent works and relevant cutting-edge reviews based on the comet assay with special regard to, e.g., clinical applications, nanoparticles or environmental risk assessment concludes this review. Taken together, the presented overview raises expectations to further decades of successful applications and enhancements of this excellent method.

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Section: Current Workmentioning

confidence: 99%

Comet assay: an essential tool in toxicological research

2016

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“…15 Alternatively, comets can be analyzed based on computer image analysis. 22,23 Image analysis systems for comet evaluation potentially strengthen the method by avoiding individual analyzer variation, 22 mostly for unexperienced laboratories and for very low irradiation doses (e.g., 0.1 kGy used to inhibit sprouting of potatoes, onions, and garlic). 24 Also, they allow fully quantitative discrimination between irradiated and nonirradiated samples, as well as they are able to set up standard dose-response curves, resulting in sufficiently accurate dose estimations.…”

Section: Detection Of Irradiated Foods By Biological Methodsmentioning

confidence: 99%

Chapter 15. Biological Techniques

Rodrigues¹,

Venâncio²

2017

Food Chemistry, Function and Analysis

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DNA is a large molecule particularly sensitive to ionizing radiation, which suffers several kinds of damage: fragmentation resulting from both singlestrand and double-strand breaks, denaturation of the DNA helix, crosslinking (e.g., production of thymine dimers, or between DNA and a protein) and base damage. 1-3 It causes primarily single strand breaks (SSBs) in genomic DNA, in addition to double strand breaks (DSBs) at ratios of SSB/ DSB of 20/1 to 70/1, as well as some detectable membrane damage. 4 In foods, this DNA susceptibility is the cause of death of most if not all living contaminants, such as microorganisms, insects, or parasites, 4 and is also the cause of changes in the food's DNA itself, which can reflect on various morphological and physiological features. DNA damage occurs predominantly by the indirect action of gamma rays, which interact with other atoms or molecules, particularly water, to produce reactive free radicals. 5 Cell death (defined for proliferating cells as the loss of reproductive capability) is predominantly induced by double-strand breaks in DNA, separated by not more than a few base pairs, which cannot generally be repaired by the cell. 6 Since irradiation with just 1 Gy introduces about 1000 DNA single-strand breaks and about 50 double-strand breaks per cell, 7

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“…The HepG2 cells were collected after being treated by 0, 5, 10, 15 and 20 μM Spinetoram for 12 h, then washed with PBS (pH 7.4) for 3 times to remove Spinetoram. After that, HepG2 cells were mixed with 1% agarose with low melting point at a ratio of 1:5 under 37°C to form monosuspension which was then cast upon the microscopic slide (Cetinkaya, Ercin, Özvatan, & Erel, 2016;Zhang, Luo, et al, 2016). The slide was rinsed by distilled water for 3 times after lysis, immersed in electrophoresis solution of ice alkaline (1 mM EDTA, 300 mM NaOH, with pH of 13.0) at 4°C for 15 min, then the electrophoresis was performed at 20 V (1 V/ cm) or 300 mA for 20 min (Ghassemi-Barghi, Varshosaz, Etebari, & Dehkordi, 2016).…”

Section: Alkaline Comet Assaymentioning

confidence: 99%

The cytotoxic effects of spinetoram on human HepG2 cells by inducing DNA damage and mitochondria-associated apoptosis

Zhang

Guo

Chen

et al. 2019

Food and Agricultural Immunology

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Spinetoram has been widely used in agriculture due to its target specificity. However, it has not been paid enough attention to the health threats of humans. This study aimed to evaluate the cytotoxic effects of the Spinetoram in human liver cells in vitro. We demonstrated that the viability of HepG2 cells was inhibited by spinetoram in a time-and dose-dependent manner. Intracellular biochemical assays revealed that Spinetoram-induced apoptosis of HepG2 cells was concurrent with a number of activated cell processes, including DNA damage, decrease of mitochondrial membrane potential, release of cytochrome c into the cytosol, marked activation of caspase-9/-3 and cleavage of poly (ADP-ribose) polymerase and up-regulation of Bax/Bcl-2 expression levels. These results indicated that a mitochondrialmediated pathway contributes to the apoptosis of Spinetoraminduced in HepG2 cells. In summary, this study shows that spinetoram poses a potential risk to human health by inducing DNA damage and mitochondria-associated apoptosis.

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Quantification of applied dose in irradiated citrus fruits by DNA Comet Assay together with image analysis

Cited by 15 publications

References 15 publications

Comet assay: an essential tool in toxicological research

Comet assay: an essential tool in toxicological research

Chapter 15. Biological Techniques

The cytotoxic effects of spinetoram on human HepG2 cells by inducing DNA damage and mitochondria-associated apoptosis

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