Using the comet and micronucleus assays for genotoxicity studies: A review

Araldi, Rodrigo Pinheiro; Melo, Thatiana Corrêa de; Mendes, Thais Biude; Júnior, Paulo Luiz de Sá; Nozima, Bruno Heidi Nakano; Ito, Eliana Tiemi; Carvalho, Rogério Falleiros; Souza, Edislane Barreiros de

doi:10.1016/j.biopha.2015.04.004

Cited by 190 publications

(160 citation statements)

References 124 publications

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“…34 The micronuclear test being the most sensitive and the most powerful statistical tool to evaluate DNA breaks, which indicate potential mutations. 35 Meanwhile, Results showed that three types of honey, which used in this study formed by different feed sources (HS, HW and HF, had no significant effect on the induction of micronuclei in bone marrow cells, which showed nearly the same values of control as shown in Table 1. Significantly (P < 0.01) as compared with negative control, especially in the low dose, although, the high dose (600 mg/kg) of honey (HS) showed significant differences (P < 0.01) of frequencies micronuclei as compared with negative control, but no statistically significant in micronucleus in comparison with positive group.…”

Section: Resultsmentioning

confidence: 64%

Investigation of role magnetized water used in supplementary feeding for honeybees to modulate the genotoxic side effects induced by cyclophosphamide in mice bone marrow cells

Farhan¹,

Chechan

Alkinani

2017

J. contemp. med. sci.

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Objective To evaluate the possible protective role of honey formed through innovative way by using magnetized water instead of tab water in supplementary feeding for bees (HW) and compared with two honey types formed by two different feeding solutions; sugar syrup (HS) and nectar of flowers (HF) against cyclophosphamide (CP) genotoxicity in mice bone marrow cells, by using the micronucleus assay (MN). Methods Mice were divided into 8 groups from 3 animals each. G1 as control. G2 were exposed to CP (40 mg/kg), G3, G4, and G5 received two dose of three types of honey at doses (300, 600 mg/kg) respectively. The other groups G6, G7 and G8 were supplemented with three types honey (300 mg/kg) in three different experimental protocols, as pre 2 h, post 2 h, and concomitant treatment for 7 and 14 days. Results Examination and analysis of MN showed no mutagenic effect of three types honey per se doses, especially in low dose (300 mg/kg). Meanwhile, CP induced a significant (P < 0.01) increase in MN frequency. While dual treatment with groups of honey HW caused a significant reduction in MN induced by CP in bone marrow cells in a time-dependant manner. Also, the results confirmed the protective efficacy of HW group and/or HF group as compared with HS group, against CP-toxicity. Conclusion Our study suggests using magnetized water in supplementary feeding of bee, that could give the honey protective effect against genotoxicity induced by CP, it is also fosters antioxidant activity of honey constituents. Therefore, honey HW can be used as an adjuvant with chemotherapeutic agents for minimizing the genotoxic side effects of the anticancer drug CP.

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Section: Resultsmentioning

confidence: 64%

Investigation of role magnetized water used in supplementary feeding for honeybees to modulate the genotoxic side effects induced by cyclophosphamide in mice bone marrow cells

Farhan¹,

Chechan

Alkinani

2017

J. contemp. med. sci.

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“…The identity of this protein was previously verified in Araldi et al [44]. The E6 oncoprotein concentration was chosen according to the literature, since the administration of 1 μg/mL of E6 oncoprotein has been suggest as therapeutic vaccine against both BPV and HPV [45]. Statistical analysis was based on cell percentage and mean of fluorescence intensity (MFI) and stand deviation, since the mean of fluorescence is the most indicated method to analyze the expression of one specific marker in absolute terms.…”

Section: Mitochondrial Membrane Potential (δψM) Analysismentioning

confidence: 98%

“…Comet assay was performed according to alkaline technique [45] in order to evaluate DNA breaks (clastogenesis) along of the six cell passages . An aliquot of 10 µL (0.1 x 105 cells) of the cells suspension was washed in PBS.…”

Section: Comet Assaymentioning

confidence: 99%

“…Material was transferred to 0.2 mL polypropylene tubes and mixed with 75 µL of low melting-point agarose (0.7% in PBS) at 37ºC. This LMA concentration do not affect the DNA migration (45). Cell suspension was transferred for 76 x 26mm microscope slides precoated with normal melting-point agarose (1.5% in PBS) at 60ºC.…”

Section: Comet Assaymentioning

confidence: 99%

“…The number of nucleoids observed per class was multiplied to the class value, resulting in a comet score. Statistical analyses were performed by the Kruskal-Wallis test, followed by the Dunn test, both with 5% of significance level as proposed by Araldi et al [45]. Linear regressions were used to evaluate the cell manipulation interference in clastogenesis.…”

Section: Comet Assaymentioning

confidence: 99%

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Primary Cultures Derived From Bovine Papillomavirus-Infected Lesions As Model To Study Metabolic Deregulation

Stocco¹

2016

Journal of Cancer Research and Therapeutic Oncology

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Bovine papillomavirus (BPV) is the etiological agent of bovine papillomatosis, disease characterized by the presence of multiple papillomas that can regress or to progress to malignances. Due to the pathological similarities with the human papillomavirus (HPV), BPV is considered a prototype to study the papillomavirus-associated oncogenic process. Although it is clear that both BPV and HPV can interact with host chromatin, the interaction of these viruses with cell metabolism remains understudied due to the little attention given to primary cultures derived from papillomavirus-infected lesions. Thus, this study analyzed the energy metabolism, including the mitochondrial membrane potential (ΔΨm) and Reactive Oxygen Species (ROS) of cells derived from cutaneous papilloma, fibropapilloma and Esophageal Carcinoma (EC) as model to evaluate the cell metabolism. These cells were cultivated until sixth passage and subjected to BPV DNA sequences identification by PCR using specific primers to BPV-1, 2 and 4. PCR results showed the presence and maintenance of at least one BPV type along the six passages analyzed. Cells derived from normal skin, without BPV DNA sequences were used as control. Results of energy metabolism showed the loss of ΔΨm in fibropapilloma and EC cells, suggesting a metabolic switch compatible to the activation of aerobic glycolysis. Cutaneous papilloma and normal skin cells showed the maintenance of ΔΨm. Paradoxically, cutaneous papilloma and fibropapilloma presented high levels of ROS production, while the EC cells reduced the ROS levels, reinforcing the activation of glycolytic metabolism. Our results suggest that the metabolic switch is mediated by BPV E6 oncoprotein, since the addition of this oncoprotein in normal cells promoted the oxidative stress. The oxidative stress showed able to activate the STAT3 nuclear factor in papilloma and fibropapilloma cells, contributing to metabolic deregulation. These data suggest that primary cultures are useful model to study the interaction between BPV and cell metabolism.

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Cytotoxicity of Metal‐Based Nanoparticles: From Mechanisms and Methods of Evaluation to Pathological Manifestations

Xiong

Xiao-hong

et al. 2022

Advanced Science

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Metal-based nanoparticles (NPs) are particularly important tools in tissue engineering-, drug carrier-, interventional therapy-, and biobased technologies. However, their complex and varied migration and transformation pathways, as well as their continuous accumulation in closed biological systems, cause various unpredictable toxic effects that threaten human and ecosystem health. Considerable experimental and theoretical efforts have been made toward understanding these cytotoxic effects, though more research on metal-based NPs integrated with clinical medicine is required. This review summarizes the mechanisms and evaluation methods of cytotoxicity and provides an in-depth analysis of the typical effects generated in the nervous, immune, reproductive, and genetic systems. In addition, the challenges and opportunities are discussed to enhance future investigations on safer metal-based NPs for practical commercial adoption.

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Using the comet and micronucleus assays for genotoxicity studies: A review

Cited by 190 publications

References 124 publications

Investigation of role magnetized water used in supplementary feeding for honeybees to modulate the genotoxic side effects induced by cyclophosphamide in mice bone marrow cells

Investigation of role magnetized water used in supplementary feeding for honeybees to modulate the genotoxic side effects induced by cyclophosphamide in mice bone marrow cells

Primary Cultures Derived From Bovine Papillomavirus-Infected Lesions As Model To Study Metabolic Deregulation

Cytotoxicity of Metal‐Based Nanoparticles: From Mechanisms and Methods of Evaluation to Pathological Manifestations

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