In vitro assessment of anticytotoxic and antigenotoxic effects of CANOVA®
Henrique Fonseca Sousa do NascimentoPlínio Cerqueira dos Santos CardosoHelem Ferreira RibeiroTatiane Cristina MotaLorena Monteiro GomesAndré Salim KhayatAdriana Costa GuimarãesMarúcia Irena Medeiros AmorimRommel Mário Rodríguez BurbanoMarcelo de Oliveira Bahia
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Abstract:
CANOVA(®) (CA) is a homeopathic immunomodulator. It contains several homeopathic medicines prepares according to the Brazilian Pharmacopoeia. CA is indicated in clinical conditions in which the immune system is impaired and against tumors. N-methyl-N-nitrosourea (NMU) is an N-nitroso compound, with genotoxic/mutagenic properties. Although several studies have shown promising results in the use of CA, there are no studies reporting possible antigenotoxic effects.This study evaluated the in vitro antigenotoxic and anticytotoxic effects of CA in human lymphocytes exposed to NMU. Samples of human lymphocytes that were subjected to different concentrations of a mixture containing CA and NMU were used. The genotoxicity/antigenotoxicity of CA was evaluated by the comet assay, anticytotoxicity was assessed by quantification of apoptosis and necrosis using acridine orange/ethidium bromide.CA significantly reduced DNA damage induced by NMU and reduced significantly the frequency of NMU-induced apoptosis after 24 h of treatment.CA has an important cytoprotective effect significantly reducing the DNA damage and apoptosis induced by the carcinogen NMU.Keywords:
Acridine orange
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Ethidium bromide
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The plasmid eliminating abilities of acridine orange, ethidium bromide and sodium dodecyl sulfate were investigated on multi drug resistant Escherichia coli from urinary tract infection specimens. Three different concentrations of each curing agent (Et-Br, SDS and AO) were used. The frequencies of cured cells were 5.55 % (with 50 μg/ml) and 11.76 % (with 75 μg/ml) for acridine orange, 14.29 % (with 100 μg/ml), 21.05 % (with 100 μg/ ml), 17.65 % (with 125 μg/ml) for ethidium bromide and 7.4 % (with 10 % w/v) & 6.67 % (with 10 % w/v) for sodium dodecyl sulfate. However, no cured cells were obtained from 100 μg/ml acridine orange, 75 μg/ml ethidium bromide and 8 and 12 % SDS. Analysis of profiles of wild type and plasmid cured strains by electrophoresis yielded bands of varying sizes for wild type cells, but none were obtained for Et-Br cured cells. Acridine orange treated cells could eliminate only plasmids of 2.7 MDa and another smaller than 2 MDa. Key Words: Plasmid curing; Escherichia coli; Ethidium Bromide; Sodium Dodecyl Sulfate; Acridine Orange. DOI: http://dx.doi.org/10.3329/bjm.v27i1.9165 BJM 2010; 27(1): 28-31
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Shailaja Kasibhatla, Gustavo P. Amarante-Mendes, Deborah Finucane,Thomas Brunner, Ella Bossy-Wetzel and Douglas R. GreenThis protocol was adapted from “Apoptosis Assays,” Chapter 15, in Cells (eds. Spector etal.). Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY, USA, 1998. Thisthree-volume set is now out of print; however, some of the microscopy methods wererepublished in Basic Methods in Microscopy, by David L. Spector and Robert D.Goldman.
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[Objective] To study the genotoxicity and cytotoxicity in root tip cells of maize with different concentration of [Methods] This study was designed to assess the potential induction of micronuclei by Ce(SO4)2 with different concentrations in the root tip cells of maize(Zea mays L). [Results] The results showed that the frequency of micronuclei(FMN) increased significantly at the concentration of Ce(SO4)2 over 10 mg/L and the mitosis index(MI) decreased when the concentration of Ce(SO4)2 reached 1000 mg/L. [Conclusion] It suggests that Ce(SO4)2 has genotoxicity and cytotoxicity to root tip cells of maize.
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Acridine orange enhances the uptake of [3H]actinomycin D in disrupted and intact human lymphocytes, as measured by liquid scintillation and autoradiography. Proflavine, quinacrine, chloroquine, and ethidium bromide are not effective. In mice, acridine orange increases the capacity of actinomycin to reduce spleen weight. Type II acridine binding to DNA may be a prerequisite for actinomycin enhancement.
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Ethidium bromide
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Total direct and direct viable counts of fresh and injured cultures of Escherichia coli were determined by image analysis in preparations stained with acridine orange, ethidium bromide and 4′,6‐diamidino‐2‐phenyl indole (DAPI). Cells stained with DAPI were not detected by image analysis. Fresh cultures stained with acridine orange or ethidium bromide gave comparable counts. Injured E. coli stained with ethidium bromide gave higher counts that with acridine orange. Injured cultures stained with acridine orange contain high proportions of green cells which are less easily detected than red cells in image analysis. In certain cases it may be better to use ethidium bromide, which stains all cells red, for direct viable counts by image analysis.
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In this paper, the genotoxicity of (1-(4-chlorophenyl)-3-(2,6-difluoro-benzoyl)urea) is investigated by a micronuclear test on Danio rerio, as a standard test object, at concentrations of 0.5, 1 and 2 mg/l. As a result of the work, a significant increase in the frequency of occurrence of micronuclei (0.73%) was found, while other nuclear anomalies in the maximum concentrations of erythrocytes were also significant. It was found that the frequency of micronuclei in concentrations of 0.5 and 1 mg/l on the fifth day of the experiment was the maximum, while at the maximum concentration (2 mg/l) the level of micronuclei was lower, which is probably due to toxic effects. An increase in the level of micronuclei may be associated with the genotoxic effect of DFB decay products. The genotoxicity results obtained using the micronucleus test method were contradictory. For this reason, it is necessary to conduct additional studies using the comet method or experiments on cell cultures.
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