• Environmental Analysis Health and Toxicology
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• 제14권1_2호
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• pp.1-12
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• 1999

Recently, several new methods for the detection of genetic damages in vitro and in vivo based on molecular biological techniques were introduced according to the rapid progress in toxicology combined with cellular and molecular biology. Among these methods, mouse lymphoma thymidine kanase (tk) gene forward mutation assay, single cell gel electrophoresis (comet assay) and transgenic animal and cell line model as a target gene of lac I (Big Blue) and lac Z (Muta Mouse) gene mutation are newly introduced based on molecular toxicological approaches. The mouse lymphoma tk$\^$+/-/ gene assay (MOLY) using L5178Y tk$\^$+/-/ mouse lymphoma cell line is one of the mammalian forward mutation assays, and has many advantages and more sensitive than hprt assay. The target gene of MOLY is a heterozygous tk$\^$+/-/ gene located in 11 chromosome, so it is able to detect the wide range of genetic changes like point mutation, deletion, rearrangement, and mitotic recombination within tk gene or deletion of entire chromosome 11. The comet assay is a rapid, simple, visual and sensitive technique for measuring and analysing DNA breakages in mammalian cells, Also, transgenic animal and cell line models, which have exogenous DNA incorporated into their genome, carry recoverable shuttle vector containing reporter genes to assess endogenous effects or alteration in specific genes related to disease process, are powerful tools to study the mechanism of mutation in vivo and in vitro, respectively. Also in vivo acridine orange supravital staining micronucleus assay by using mouse peripheral reticulocytes was introduced as an alternative of bone marrow micronucleus assay. In this respect, there was an International workshop on genotoxicity procedure (IWGTP) supported by OECD and EMS (Environmental Mutagen Society) at Washington D. C. in March 25-26, 1999. The objective of IWGTP is to harmonize the testing procedures internationally, and to extend to finalization of OECD guideline, and to the agreement of new guidelines under the International Conference of Harmonization (ICH) for these methods mentioned above. Therefore, we introduce and review the principle, detailed procedure, and application of MOLY, comet assay, transgenic mutagenesis assay and supravital staining micronucleus assay.

• 한국식품위생안전성학회지
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• 제20권1호
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• pp.58-63
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• 2005

시험물질 산삼배양추출물의 유전독성 평가를 위해 수컷 ICR 마우스 골수세포를 이용한 소핵시험을 실시하였다. 1 회 투여 최고량은 예비 시험에서 결정하였다. 약 7주령의 수컷 마우스에 시험물질 0, 500, 1,000 및 2,000 mg/kg의 용량을 1일 1회 2일간 복강내 투여하고, 최종 투여로부터 약 24시간 후에 골수세포를 수거하여 소핵 유발과 세포독성을 평가하였다. 개체당 2,000개의 다염성적혈구(michromatic erythrocyte, PCE)중에 나타나는 소핵을 가진 다염성적혈구(micronucleated polychromatic erythrocyte, MNPCE)의 수를 계수한 결과, 모든 시험물질 투여군은 음성 대조군에 비해 통계학적으로 유의한 증가는 나타나지 않았으며 일반 증상에서도 모든 시험군은 투여로 인한 것으로 판단되는 증상은 관찰되지 않았다. 부검시 체중에 있어서는 시험물질 최고 용량군에서 유의한 감소가 관찰되었다. 따라서 산삼배양추출물은 위 시험 조건에서, 본 시험에 사용한 마우스 골수세포에 소핵을 유발하지 않는 것으로 사료된다.

• 한국환경성돌연변이발암원학회지
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• 제21권1호
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• pp.51-56
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• 2001

Kimchi is a major Korean traditional fermented food, as a supplying source of vitamin and minerals which is prepared with various vegetables and condiments such as red pepper, garlic and salted fish etc. There are many types of Kimchi depending on the ingredients and preparation methods used. To investigate the clastogenicity and anticlastogenicity of Baechu (Chinese cabbage) Kimchi and Buchu (leek, Allium odorum) Kimchi in mouse, it was performed acridine orange supravital staining of micronucleus (AOSS-MN) assay using mouse peripheral reticulocytes. Baechu Kimchi and Buchu Kimchi were cultivated by organic agricultural technique, and Kimchi samples were prepared by methanol extraction and lyophilization. First of all, it was studied the clastogenicity of two Kimchi samples themselves (250-1,000 mg/kg) after oral adminstration in mouse. And also to study the anticlastogenic effect of oral administration of Kimchi samples, mitomycin C (MMC, 1 mg/kg, i.p.) was used as micronucleus inducing agent in this study. Dosing scheme was performed as simultaneous (co-treatment), 3 hr before (pre-treatment) and 3 hr after (post-treatment) with MMC treatment. Two Kimchi samples in the range of 250-1,000 mg/kg did not reveal any clastogenic effect in AOSS-MN assay in mouse. They also revealed anticlastogenic effects in post-treatment of Baechu Kimchi (1,000 mg/kg), and in pre-treatment of Buchu Kimchi (500 and 1,000 mg/kg) with statistical significance. The anticlastogenic effect revealed 1 and 6 hr after treatment of Baechu Kimchi, and Buchu Kimchi with 3 and 6 hr pretreatment. Consequently, it is suggested that antimutagenic and anticlastogenic mechanisms of Baechu and Buchu Kimchi in vivo attributed to sipindle formation and kinetic behavior of mutagens such as absorption and metabolism etc.

• 한국한의학연구원논문집
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• 제18권2호
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• pp.151-157
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• 2012

In this research, the genotoxic effect of Bupleuri Radix (BR), the dried roots of Bupleurum falcatum Linne has been traditionally used as anti-inflammatory agent, was evaluated using the mouse micronucleus test. BR aqueous extract (yield = 16.52%) was administered once a day for 2 continuous days by oral gavage to male ICR mice at doses of 2,000, 1,000 and 500 mg/kg. Cyclophosphamide (CPA) 70 mg/kg was used as a known genotoxic agent in a positive control. The appearance of a micronucleus (MN) in polychromatic erythrocyte (PCE) is used as an index for genotoxic potential, and PCE ratio is used as an index of cytotoxicity. Although significant (p<0.01) increase of the number of PCE with one or more nuclei (MNPCE) was detected in CPA treated groups, no significant increases of MNPCE numbers were observed in all three different dosages of BR extracts treated mice with over 0.30 of the individual polychromatic erythrocyte ratio in all mice used in this study. The results obtained indicated that BR extract shows no genotoxicity effects up to 2,000 mg/kg dosing levels the limit dosage in rodents.

• 대한예방한의학회지
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• 제16권1호
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• pp.81-89
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• 2012

Objectives : In this research, the genotoxic effect of $Scutellariae$ $Radix$(SR), the dried roots of $Scutellaria$ $baicalensis$ Georgi has been traditionally used as antipyretic agent, was evaluated using the mouse micronucleus test. Methods : SR aqueous extract(yield = 27.2%) was administered once a day for 2 continuous days by oral gavage to male ICR mice at doses of 2,000, 1,000 and 500 mg/kg. Cyclophosphamide(CPA) 70 mg/kg was used as a known genotoxic agent in a positive control. The appearance of a micronucleus(MN) in polychromatic erythrocyte(PCE) is used as an index for genotoxic potential, and PCE ratio is used as an index of cytotoxicity. Results and Conclusions : Although significant(p<0.01) increase of the number of PCE with one or more nuclei(MNPCE) was detected in CPA treated groups, no significant increases of MNPCE numbers were observed in all three different dosages of SR extracts treated mice with over 0.33 of the individual polychromatic erythrocyte ratio in all mice used in this study. The results obtained indicated that SR extract shows no genotoxicity effects up to 2,000 mg/kg dosing levels - the limit dosage in rodents.

• Environmental Analysis Health and Toxicology
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• 제24권2호
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• pp.127-136
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• 2009

We have investigated the genotoxicity of 3 chemicals, aluminum oxide, calcium oxide, sodium tetraborate using mammalian erythrocyte with micronucleus induction. It was performed using 9 week male ICR mice. At 24 hours after treatment with 3 chemicals with oral route, mice were sacrificed and bone marrow cells were prepared for smear slides. As a result of counting the micronucleated polychromatic erythrocyte (MNPCE) of 2,000 polychromatic erythrocytes (PCE), all treatment groups did not show statistically significant increase than negative control group. And there was no clinical sign related with injection of the 3 chemicals. It was concluded that the 3 chemicals did not induce micronucleus in the bone marrow cells of ICR mice, and these results indicate that the 3 chemicals have no mutagenic potential under the condition in each studies.

• Toxicological Research
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• 제24권4호
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• pp.321-328
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• 2008

Smoke flavors based on the thermal decomposition of wood have been applied to a variety of food products as an alternative for traditional smoking. Despite its increasing use, the available genotoxicity data on wood smoke flavors (WSF) are still controversial. Thus, potential genotoxic effects of WSF in four short-term in vitro genotoxicity assays were investigated, which included the Ames assay, chromosomal aberration assay, micronucleus test and the alkaline comet assay. WSF did not cause any mutation in the Ames assay using five tester strains at six concentrations of 0.16, 0.31, 0.63, 1.25, 2.5 and 5 ${\mu}l/plate$. To assess clastogenic effect, the in vitro chromosomal aberration assay was performed using Chinese hamster lung cells. No statistically significant increase in the number of metaphases with structural aberrations was observed at the concentrations of 1.25, 2.5, and 5 ${\mu}l/ml$. The in vitro comet assay and micronucleus test results obtained on L5178Y cells also revealed that WSF has no genotoxicity potential, although there was a marginal increase in micronuclei frequencies and DNA damage in the respective micronucleus and comet assays. Taken together, based on the results obtained from these four in vitro studies, it is concluded that WSF is not a mutagenic agent in bacterial cells and causes no chromosomal and DNA damage in mammalian cells in vitro.

• 대한한방종양학회지
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• 제7권1호
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• pp.99-107
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• 2001

In the present studies, decursinol angelate, decursin isolated from Angelica gignatis radix and oil fraction of Cnidii rhizoma was analyzed by normal phase HPLC and GC/MS respectively. The standardized water extracts of Angelica gignatis radix, Cnidii rhizoma and its complex named Gung-gui-tang was tested the anti mutagenic effects by in vitro genotoxicity using Salmonella reversion assay (Ames test) and micronucleus test in chinese hamster ovary(CHO) cells. Angelica gignatis radix, Cnidii rhizoma and Gung-gui-tang was not exhibited the antimutagenic effects in the Salmonella reversion assays with or without metabolic activation. However, the micronucleus test assays, Angelica gignatis radix and Gung-gui-tang was showed the antimutagenic effects significantly. The maximum inhibition observed with Gung-gui-tang was reduced by 59% in the micronucleus test without metabolic activation. In this paper, results are presented on the availability of potential antimutagenic activity of the water extracts of Gung-gui-tang.

• Asian Pacific Journal of Cancer Prevention
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• 제14권9호
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• pp.5257-5261
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• 2013

The micronucleus frequency (MNF) in peripheral blood lymphocytes (PBL) is a biomarker of chromosomal damage and genome instability in human populations.The relationship of micronucleus frequency with prognosis of patients with acute leukemia is not clear. We therefore investigated MNF in mitogen-activated peripheral blood lymphocytes from patients with hematologic diseases and solid tumours. Patients included 50 with acute leukemia, 49 diagnosed with myelodysplastic syndrome (MDS), 54 with benign blood diseases, and 45 with solid tumours, examined with 50 healthy controls. The mean MNF was significantly higher in cases of hematologic diseases and solid tumor patients than in healthy controls (P<0.001). There was no evident difference between MNF in the acute leukemia ($7.15{\pm}2.18$) and solid tumor groups ($7.11{\pm}1.47$), but both were higher than in the MDS group ($5.12{\pm}1.29$) and benign blood diseases group ($3.08{\pm}1.08$). Taking 7.15‰, the average MNF of the acute leukemia group as standard, and dividing 50 cases of acute leukemia patients into high MNF group ($MNF{\geq}7.15$‰) and low MNF group (MNF<7.15‰). The overall response (complete remission + partial remission) rates of the low MNF group were significantly higher than in the high MNF group (P=0.001). The high MNF group further showed lower overall survival rates than the low MNF group. MNF expression and progression-free survival seemed to have a opposite relationship, with a correlation coefficient of -0.702. These data indicate that MNF in peripheral blood lymphocytes is important for evaluation of prognosis of acute leukemia patients, and it can reflect progression of disease to a certain degree.

• Journal of Applied Biological Chemistry
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• 제62권4호
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• pp.355-359
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• 2019

In vivo micronucleus tests were performed to investigate the mutagenic potential of 4-butylaniline and N-butylaniline, which are used in dye intermediates and organic intermediates respectively. Groups of 5 male ICR mice were treated with vehicle or 4-butylaniline for 2 consecutive days by oral gavage at concentrations of 0 (control), 64, 160, 400, and 1000 mg/kg. Statistically significant and dose-dependent increases were found for micronuclei frequencies in male mice (p <0.05). These results suggest that 4-butylaniline can induce genetic effects in the micronuclei of male mouse bone marrow cells. Based on the positive results obtained in cytogenetic analyses of somatic cells in vivo, Globally Harmonized System of Classification and Labelling of Chemicals Category 2 was assigned. N-butylaniline was administered for 2 consecutive days by oral gavage to male ICR mice at dose of 0 (control), 64, 160, 400, and 800 mg/kg. N-butylaniline tested negative for micronuclei induction in mice, although N-butylaniline was associated with micronucleus induction at the highest dose. Based on the negative results obtained for cytogenetic analyses of somatic cells in vivo, "Not Classified" was assigned.