• The Korea Journal of Herbology
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• v.25 no.2
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• pp.137-144
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• 2010

Objectives : We investigated genetic toxicity of HMCO5 using the Micronucleus Test in bone marrow cells of mice and Bacterial Reverse Mutation Assay in plate incorporation method according to OECD Guidelines and KFDA Guidelines. Methods : 1. Micronucleus test: The male rats were divided into 5 groups, respectively; G(1), treated with distilled water: G(2), treated with 1250mg/kg HMC05: G(3), treated with 2500mg/kg HMC05, G(4), treated with 5,000mg/kg HMC05; G(5), treated with Cyclophosphamide $H_2O$. Sterilized distilled water and HMC05 were administered for two consecutive days. Cyclophosphamide $H_2O$ was administered once on the day of 2nd administration. 2. Bacterial Reverse Mutation Aassay: Experimental groups were divided into two groups: with S-9mix(+S) or without S-9mix(-S). Each group treated with sterilized distilled water only, HMCO5(62, 185, 556, 1,667, $5,000{\mu}g$/plate) and, positive vehicles(Sodium azide, 2-Aminoanthracene, 4-Nitroquinoline N-oxide, ICR 191), respectively. Results : HMC05 did not show any changes in the number of micronucleated polychromatic erythrocytes(MNPCE) among 200 polychromatic erythrocytes compare to negative control. However, there were significant (p<0.01) increase with CPA in MNPCE. In Bacterial Reverse Mutation Aassay, no significant increases in the number of revertant colonies compared to (삭제) negative control were detected in all concentrations of HMC05. Conclusions : These results indicate that HMC05 did not show any genotoxicity against in Micronucleus test and Bacterial Reverse Mutation Aassay.

• Herbal Formula Science
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• v.14 no.1
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• pp.141-167
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• 2006

The genotoxicity of water extract of Bojungikkeehapdaechilki-tang was tested by In Vitro Chromosome Aberration Test. Bacterial Reverse Mutation Assay and Micronucleus test according to OECD Guidelines and KFDA Guidelines. The obtained results were as follows : 1. Chromosome Aberration Test: In Vitro Chromosome Aberration Test of Bojungikkeehapdaechilki-tang extracts was carried out using cultured Chinese hamster lung cells in the presence and absence of metabolic activation system(S-9 mix). No significant changes in the number of aberrant metaphases having structural and number of aberrations were detected in Bojungikkeehapdaechilki-tang extracts treated groups. 2. Bacterial Reveres Mutation Assay: Bojungikkeehapdaechilki-tang extracts was evaluated for its potential to induce reverse mutation in the histidine auxotroph strains of Salmonella typhimurium such as TA100, TA1535, TA98 and TAl537 and the tryptophan auxotroph strain of Escherichia coli WP2 uvrA. No significant changes in the number of revertant colonies compared to its negative control were detected in Bojungikkeehapdaechilki-tang extracts treated groups against all 5 strains. 3. Micronucleus test: Micronucleus test of Bojungikkeehapdaechilki-tang extracts were performed using specific pathogen free 7-week old male ICR mouse. No significant changes in the number of micronucleated polychromatic erythrocytes among 2000 polychromatic erythrocytes compared to negative control were detected in all Bojungikkeehapdaechilki-tang extracts treated groups. In summarized above-mentioned results, it is concluded that Bojungikkeehapdaechilki-tang extracts have not genotoxicity against In Vitro Chromosome Aberration Test, Bacterial Reverse Mutation Assay and Micronucleus test.

• The Korea Journal of Herbology
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• v.22 no.4
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• pp.287-300
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• 2007

Objectives : The genotoxicity of extract of "Hyeonggaeyeongyotang", a polyherbal formula has been used as a tonic agents in oriental medicine was tested. Methods : Extract of "Hyeonggaeyeongyotang" was tested by In Vitro Chromosome Aberration Test, Bacterial Reverse Mutation Assay and Micronucleus test according to OECD Guidelines and KFDA Guidelines [2005-60]. Results : The obtained results were as follows: 1. Chromosome Aberration Test: No significant changes in the number of aberrant metaphases having structural and number of aberrations were detected in all concentrations of "Hyeonggaeyeongyotang" extracts treated in this study. 2. Bacterial Reverse Mutation Assay: No significant increases in the number of revertant colonies compared to its negative control were detected in all concentrations of "Hyeonggaeyeongyotang" extracts treated in this study against all 5 strains except for $50{\mu}g/ml$ treated group where significantly decreases in colony numbers were detected agains all five strains used in this study as pharmacological effects not genotoxicity. 3. Micronucleus test: No significant changes in the number of micronucleated polychromatic erythrocytes among 2000 polychromatic erythrocytes compared to negative control were detected in all "Hyeonggaeyeongyotang" extracts-dosing groups tested. Conclusions : From above-mentioned results, it is concluded that "Hyeonggaeyeongyotang" extracts have not any genotoxicity against In Vitro Chromosome Aberration Test, Bacterial Reverse Mutation Assay and Micronucleus test.

• Environmental Mutagens and Carcinogens
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• v.22 no.4
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• pp.319-323
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• 2002

In order to investigate the safety of Aralia elata extract causing the reduction in the blood glucose level and oxidative stress in diabetes animals, these genotoxicity studies in bacterial and mammalian cell assay system such as Ames bacterial reverse mutation test and chromosomal aberration assay were performed. As results, in Ames bacterial reversion assay the extract in the range of 5,000-625 ug/plate did not induce mutagenicity in Salmonella typhimurium TA 98, TA 100, TA 1535 and TA 1537 strains with and without metabolic activation of S-9 mixture. For chromosomal aberration assay, $IC_{50}$ (50% inhibition concentration of cell growth) of the extract were determined; 792 $\mu\textrm{g}$/$m\ell$ without and 524 $\mu\textrm{g}$/$m\ell$ with S-9 mixture in Chinese hamster lung (CHL) fibroblast cell culture. Any significant chromosomal aberration was not observed in CHL cells treated with the extract at the concentrations of 792, 396 and 198 $\mu\textrm{g}$/$m\ell$ or 524, 262 and 131 $\mu\textrm{g}$/$m\ell$ in the absence or presence of S-9 metabolic activation, respectively. From these results, Aralia elata extract did not induce any harmful effects on the gene in bacteria and mammalian cell system used in these experiments.

• Molecular & Cellular Toxicology
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• v.3 no.1
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• pp.53-59
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• 2007

This study was conducted to evaluate the mutagenic potential of dimethyl isophthalate (DMIP) using Ames bacterial reverse mutation test, chromosomal aberration test and mouse lymphoma $tk^{+/-}$ gene assay. As results, in Ames bacterial reversion assay, DMIP was tested up to the concentration of 5,000 ${\mu}g$/plate and did not induce mutagenicity in Salmonella typhimurium strains TA98, TA100, TA1535 and TA1537, and Escherichia coli WP2uvrA with or without metabolic activation (S9 mix). Using cytotoxicity test, the maximal doses of DMIP for chromosomal aberration assay were determined at 1,250 ${\mu}g/mL$, which was a minimum precipitation concentration ($IC_{50}>1,940\;{\mu}g/mL$ or 10 mM) and at 155 ${\mu}g/mL$ ($IC_{50}:155\;{\mu}g/mL$) in the presence and the absence, respectively, of S9 mix. DMIP in the presence of S9 mix induced statistically significant (P<0.001) increases in the number of cells with chromosome aberrations at the dose levels of over 250 ${\mu}g/mL$, when compared with the negative control. However, DMIP in the absence of S9 mix did not caused significant induction in chromosomal aberrant cells. In MLA, DMIP at the dose range of 242.5-1,940 ${\mu}g/mL$ in the presence of S9 mix induced statistically significant increases in mutation frequencies related to small colony growth, whereas any significant mutation frequency was not observed in absence of S9 mix. From these results, it is conclusively suggested that dimethyl isophthalate may be a clastogen rather than a point mutagen.

• Molecular & Cellular Toxicology
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• v.2 no.3
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• pp.151-158
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• 2006

Khal was a synthetic congener of halocidin, a heterodimeric peptide consisting of 19 and 15 amino acid residues detected in Halocynthia aurantium. This compound was considered a candidate for the development of a novel peptide antibiotic. The genotoxicity of Khal was subjected to high throughput toxicity screening (HTTS) because they revealed strong antibacterial effects. Mouse lymphoma thymidine kinase ($tk^{+/-}$) gene assay (MOLY), single cell gel electrophoresis (Comet) assay and chromosomal aberration assay in mammalian cells and Ames reverse mutation assay in bacterial system were used as simplified, inexpensive, short-term in vitro screening tests in our laboratory. These compounds are not mutagenic in S. typhimurium TA98 and TA100 strains both in the presence and absence of metabolic activation. Before performing the comet assay, $IC_{20}$ of Khal was determined the concentration of $25.51\;{\mu}/mL\;and\;21.99\;{\mu}g/mL$ with and without S-9, respectively. In the comet assay, Khal was not induced DNA damage in mouse lymphoma cell line. Also, the mutation frequencies in the Khal-treated cultures were similar to the vehicle controls. It is suggests that Khal is non-mutagenic in MOLY assay. And no clastogenicity was observed in Khal-treated Chinese hamster lung cells. The results of this battery of assays indicate that Khal has no genotoxic potential in bacterial or mammalian cell systems. Therefore, we suggest that Khal, as the optimal candidates with both no genotoxic potential and antibacterial effects must be chosen.

• Toxicological Research
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• v.29 no.3
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• pp.217-219
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• 2013

Hypertension is a serious health problem due to high frequency and concomitant other diseases including cardiovascular and renal dysfunction. Olmesartan cilexetil is a new antihypertensive drug associated with angiotensin II receptor antagonist. This study was conducted to evaluate the mutagenicity of olmesartan cilexetil by bacterial reverse mutation test using Salmonella typhimurium (TA100, TA1535, TA98, and TA1537) and Escherichia coli (WP2 uvrA). At the concentrations of 0, 62, 185, 556, 1667, and 5000 ${\mu}g$/plate, olmesartan cilexetil was negative in both Salmonella typhimurium and Escherichia coli regardless of presence or absence of metabolic activation system (S9 mix). These results demonstrate that olmesartan cilexetil does not induce bacterial reverse mutation.

• Toxicological Research
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• v.20 no.1
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• pp.43-47
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• 2004

In order to investigate the safety of a water extract (ADP) of 1 : 1 mixture of Anemarrhena rhizoma and Phellodendron cortex for alleviating benign prostate hyperplasia, genotoxicity studies in bacterial and mammalian cell assay systems, namely, the Ames bacterial reverse mutation and chromosomal aberration assays were performed. As shown by the results of the Ames bacterial reversion assay, ADP in the range of 625-5000 $\mu\textrm{g}$/plate did not induce mutagenicity in Salmonella typhimurium TA 98, TA 100, TA 1535 and TA 1537 strains in the absence or in the presence of S9 (the microsomal fraction of rat liver homogenate) metabolic activation. The $IC_{50}$ (50% cell growth inhibition concentration) values of ADP for the chromosomal aberration assay were determined; these were 2425 $\mu\textrm{g}$/ml in the absence and 8126 $\mu\textrm{g}$/ml in the presence of S9 metabolic activation in Chinese hamster lung (CHL) fibroblast cell culture. No chromosomal aberration was observed in CHL cells treated with ADP at 2425, 1212.5 and 606.25 $\mu\textrm{g}$/ml in the absence, or at 8126, 4063 and 2031.5 $\mu\textrm{g}$/ml in the presence of S9 metabolic activation. These results show that under the conditions used, ADP does not harmfully affect the bacterial or mammalian cell system at the gene level.

• Journal of Food Hygiene and Safety
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• v.14 no.3
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• pp.259-264
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• 1999

To evaluate the bacterial reverse mutation of xylooligosaccharide(XO)s the in vitro Ames test using Salmonella typhimurium (TA9S, TAIOO, TA1535, TA1537) and Escherichia coli (WP2 uvrA) was performed. XO was negative in Ames test with Salmonella typhimurium and Escherichia coli with and without rat liver microsomal enzyme (S-9 fraction). According to the results, XO does not cause bacterial reverse mutation.

• Molecular & Cellular Toxicology
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• v.1 no.3
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• pp.179-188
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• 2005

To develop the novel anti-allergic drug, many sophoricoside derivatives were synthesized. Among these derivatives, JSH-II-3, VI-3, VII-3, VIII-3, VII-20 and VII-20 (sodium salt) were selected and subjected to high throughput toxicity screening (HTTS) because they revealed strong IL-5 inhibitory activity and limitation of quantity. Single cell gel electrophoresis (Comet) assay, mouse lymphoma thymidine kinase ($tk^{+/-}$) gene assay (MOLY), chromosomal aberration assay in mammalian cells and Ames reverse mutation assay in bacterial system were used as simplified, inexpensive, short-term in vitro screening tests in our laboratory. Through the primary screening using the comet assay, we could choose the first candidates of sophoricoside derivatives with no genotoxic potentials as JSH-VI-3, VII-3, VII-20 and VII-20 (sodium salt). Also JSH-VII-3, VII-20 and VII-20 (sodium salt) are non-mutagenic in MOLY assay, while JSH-II-3 is mutagenic at high concentration with the presence of metabolic activation system in both comet assay and MOLY assay. The selected derivatives (JSH-VI-3, VII-3, VII-20 and VII-20 (sodium salt) are not mutagenic in S. typhimurium TA98 and TA100 strains both in the presence and absence of metabolic activation. From results of chromosomal aberration assay, 6 h treatment of JSH-VI-3, VII-3 and VII-20 (sodium salt) were not revealed clastogenicity both in the presence and absence of S-9 mixture. Therefore, we suggests that JSH-VI-3, VII-3, VII-20 and VII-20 (sodium salt), as the optimal candidates with both no genotoxic potential and IL-5 inhibitory effects must be chosen. To process the development into new anti-inflammatory drug of these derivatives, further investigation will need.