Physical principles and new applications of comet assay

Afanasieva, K. S.; Sivolob, Andrei

doi:10.1016/j.bpc.2018.04.003

Cited by 39 publications

(29 citation statements)

References 55 publications

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“…The detection of DNA damage using this method relies on the fact that DNA strand breaks facilitate DNA migration from the nucleoids towards the anode during gel electrophoresis, thereby forming a “comet tail”. Whereas the alkaline comet assay (performed at pH > 13) detects both DNA single-stranded breaks (SSBs) and DSBs, only DSBs are detected through the neutral comet assay [78,79]. Taken together, the results obtained through this method suggest that Cd exposure induces DNA strand breaks in different plant species including V. faba [80,81,82], Nicotiana tabacum [83,84], Allium sativum [82], Solanum tuberosum [85], Allium cepa [80,86,87], Lemna minor [88,89], Lactuca sativa [86,90], H. vulgare [91], Brassica oleracea and Trifolium repens [92].…”

Section: Vegetative Plant Growthmentioning

confidence: 99%

Cadmium and Plant Development: An Agony from Seed to Seed

Huybrechts

Cuypers

Deckers

et al. 2019

IJMS

148

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Anthropogenic pollution of agricultural soils with cadmium (Cd) should receive adequate attention as Cd accumulation in crops endangers human health. When Cd is present in the soil, plants are exposed to it throughout their entire life cycle. As it is a non-essential element, no specific Cd uptake mechanisms are present. Therefore, Cd enters the plant through transporters for essential elements and consequently disturbs plant growth and development. In this review, we will focus on the effects of Cd on the most important events of a plant’s life cycle covering seed germination, the vegetative phase and the reproduction phase. Within the vegetative phase, the disturbance of the cell cycle by Cd is highlighted with special emphasis on endoreduplication, DNA damage and its relation to cell death. Furthermore, we will discuss the cell wall as an important structure in retaining Cd and the ability of plants to actively modify the cell wall to increase Cd tolerance. As Cd is known to affect concentrations of reactive oxygen species (ROS) and phytohormones, special emphasis is put on the involvement of these compounds in plant developmental processes. Lastly, possible future research areas are put forward and a general conclusion is drawn, revealing that Cd is agonizing for all stages of plant development.

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Section: Vegetative Plant Growthmentioning

confidence: 99%

Cadmium and Plant Development: An Agony from Seed to Seed

Huybrechts

Cuypers

Deckers

et al. 2019

IJMS

148

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“…The comet assay, known as single cell gel electrophoresis (SCRE), helps to determine whether there has been DNA damage to a single cell from apoptosis (cell death) or cytotoxicity and the extent of this damage [27]. Cells embedded in agarose on a microscope slide are lysed with detergent and high salt to form nucleoids containing supercoiled loops of DNA linked to the nuclear matrix.…”

Section: Resultsmentioning

confidence: 99%

Cytoprotective Effect of Antioxidant Pentapeptides from the Protein Hydrolysate of Swim Bladders of Miiuy Croaker (Miichthys miiuy) against H2O2-Mediated Human Umbilical Vein Endothelial Cell (HUVEC) Injury

Shi-ying

Wang

Zhao

et al. 2019

IJMS

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In our previous research, ten antioxidant pentapeptides including FYKWP, FTGMD, GFEPY, YLPYA, FPPYERRQ, GFYAA, FSGLR, FPYLRH, VPDDD, and GIEWA were identified from the hydrolysate of miiuy croaker (Miichthys miiuy) swim bladder. In this work, their protective function on H2O2-induced oxidative damage to human umbilical vein endothelial cells (HUVECs) was studied. Results indicated that there was no significant difference in the HUVEC viability between the normal group and the treated groups with the 10 pentapeptides at the concentration of 100 μM for 24 h (p < 0.05). Furthermore, FPYLRH of 100 μg/mL extremely significantly (p < 0.001) increased the viability (80.58% ± 5.01%) of HUVECs with H2O2-induced oxidative damage compared with that of the model group. The protective mechanism indicated that FPYLRH could extremely significantly (p < 0.001) increase the levels of superoxide dismutase (SOD) (211.36 ± 8.29 U/mg prot) and GSH-Px (53.06 ± 2.34 U/mg prot) and decrease the contents of reactive oxygen species (ROS) (139.1 ± 11.8% of control), malondialdehyde (MDA) (13.66 ± 0.71 nM/mg), and nitric oxide (NO) (4.36 ± 0.32 µM/L) at the concentration of 100 μM in HUVECs with H2O2-induced oxidative damage compared with those of the model group. In addition, FPYLRH dose-dependently protected DNA in oxidative damage HUVECs model. These results suggested that FPYLRH could significantly attenuate the H2O2-induced stress injury in HUVECs and might be used as a potential natural antioxidant in the functional food industries.

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“…El ensayo cometa presenta mayor ventaja en la detección de genotóxicos, debido a la capacidad de resolver si un agente produce daños en el ADN a bajas dosis, manejo sencillo, bajo costo y reproducible (14) . De allí, su aplicación en genética toxicológica.…”

Section: Discussionunclassified

Daño en el ADN de linfocitos humanos por efecto de cloroquina

Amésquita

Cruz-Briceño

Prieto

2018

Rev Peru Med Exp Salud Publica

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Citar como: Amésquita L, Cruz-Briceño MN, Prieto Z. Daño en el ADN de linfocitos humanos por efecto de cloroquina. Rev Peru Med Exp Salud Publica. 2018;35(3):471-5. RESUMENEl objetivo del estudio fue evaluar el efecto de cloroquina (CQ) en linfocitos humanos a través del ensayo cometa. Los linfocitos fueron aislados de muestras de sangre periférica obtenidas de tres donantes sanos, no fumadores de 24 a 30 años. Los linfocitos aislados fueron expuestos durante una hora a diversos tratamientos: peróxido de hidrógeno 2,5 % (control positivo), buffer fosfato salino 1X (control negativo) y cloroquina a concentraciones de 0 µg/ml, 0,25 µg/ml; 5 µg/ml y 300 µg/ml. Se registró los promedios del porcentaje de ADN en la cola del cometa, momento de la cola y momento de la cola de Olive, encontrándose diferencias significativas entre las diversas concentraciones de CQ (p<0,01). Asimismo, la magnitud de daño del ADN se incrementó en función de la concentración de CQ. Se demostró el efecto genotóxico de CQ en linfocitos humanos expuestos in vitro. ABSTRACTThe objective of the study was to evaluate the effect of chloroquine (CQ) on human lymphocytes through the Comet Assay. Lymphocytes were isolated from peripheral blood samples obtained from three healthy, non-smoking donors aged 24 to 30 years. The isolated lymphocytes were exposed for one hour to various treatments: hydrogen peroxide 2.5% (positive control), saline buffer phosphate 1X (negative control) and chloroquine at concentrations of 0 µg/ml, 0.25 µg/ml; 5 µg/ml and 300 µg/ml. The averages of the percentage of DNA in the comet tail, moment of tail and moment of Olive's tail were recorded, with significant differences between the different concentrations of CQ (p<0.01). Also, the magnitude of DNA damage was increased as a function of the CQ concentration. The genotoxic effect of CQ was demonstrated in human lymphocytes exposed in vitro.

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Physical principles and new applications of comet assay

Cited by 39 publications

References 55 publications

Cadmium and Plant Development: An Agony from Seed to Seed

Cadmium and Plant Development: An Agony from Seed to Seed

Cytoprotective Effect of Antioxidant Pentapeptides from the Protein Hydrolysate of Swim Bladders of Miiuy Croaker (Miichthys miiuy) against H2O2-Mediated Human Umbilical Vein Endothelial Cell (HUVEC) Injury

Daño en el ADN de linfocitos humanos por efecto de cloroquina

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