• 한국약용작물학회지
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• 제12권6호
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• pp.463-472
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• 2004

한약재 향유의 정유성분, absolute 및 oleoresin을 추출분리 동정하고, 지질과산화 억제활성, 세포독성, 돌연변이 유발성 검정 및 거담효과 등의 생리활성을 측정하였다. 향유의 정유성분의 수율은 0.34%이었으며, naginate ketone이 29.37%와 elsholtizia ketone이 14.37%로 가장 많이 함유되어 있었다. 향유의 absolutes의 수율은 11.33%이었으며, methyl linolenate가 12.07%이었고, palmitic acid 10.46% 비율로 존재하고 있었다. 향유의 oleoresins의 수율은 15.24%이었으며, 9,12,15-octadecatrienoic acid가 18.62%로 가장 높은 비율을 차지하고 있었다. 세 방법의 지질과산화 억제활성시험에서 향유의 정유성분과 oleoresins을 $200\;{\mu}g/m{\ell}$의 농도로 처리했을 때 지질과산화를 잘 억제하였으나. absolutes는 같은 농도에서 거의 억제하지 못하였다. 향유의 정유성분과 oleoresins은 24시간에서 $EC_{50}$값이 $25\;{\mu}g/m{\ell}$이하로 고농도에서만 약간의 세포독성이 있으며, 향유의 absolues와 oleoresins은 고 농도에서도 돌연변이 유발성과 향균성이 나타나지 않았으나 정유성분은 $500\;{\mu}g/plate$ 이상 처리시 돌연변이 유발성 뿐만 아니라 항균성도 있는 것으로 나타났다. 향유의 정유성분과 oleoresins은 약간의 거담효과가 있는 것으로 판단된다.

• 한국식품영양과학회지
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• 제20권3호
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• pp.260-265
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• 1991

Salmonella/microsome assay system을 이용하여 N -nitrosodimethylamine (NDMA)의 돌연변이유발성을 검토하는 효과적인 방법과 비타민 C가 NDMA자체의 돌연변이유발과 nitrite와 2급아민으로부터의 NDMA 생성에 미치는 영향을 연구하였다. NDMA를 활성화시키기 위한 S9중 Aroclor1254로 induce시킨 hamster S9이 가장 효과가 있었으며 Aroclor 1254나 phenobarbital로 induce시킨 rat 59 mix에 의하여서는 활성화가 약하였다. DMSO와 ethanol에 NDMA를 녹였을 때는 돌연변이유발실험에서 저해효과가 나타났으나 phosphate buffer(pH 7. 4), 증류수, 95% methanol, Tween 80 + water(l : 4)는 저해 효과가 없었다. 비타민 C는 이미 생성환 NDMA에 대해서는 돌연변이 유발저해 효과를 나타내지 않았지만 nitrite와 2급 아민(dimethylamine)으로 부터의 NDMA 생성은 크게 저해하였다. 반응물에 비타민C를 첨가한 경우 Salmonella typhimurium TAl00의 revertant 수가 spontaneous숫자 수준으로 감소되었으며 HPLC를 이용한 NDMA의 분석에서도 거의 95% 까지의 정량적인 감소를 관찰할 수 있었다.

• 약학회지
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• 제37권1호
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• pp.18-29
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• 1993

This study was initiated to investigate the effective action and mechanism of GE-132 (Carboxyethylgermanium sesquioxide)on benzo(a)pyrene, which have strong carcinogenicity and mutagenicity. To confirm desmutagenic effect (inhibition of metabolic processes of benzo(a)pyrene with S9 Mix or inactivation of the mutagenicity of benzo(a)pyrene metabolites) and antimutagenic effect (inhibition of gene-expression of reverted genes) of GE-132 against benzo(a)pyrene using with Salmonella typhimuyium TA98 Ames test was performed. The revertants in desmutagenicity test were decreased significantly in the combined groups of benzo(a)pyrene and GE-132 than benzo(a)pyrene only, without inhibition the metabolism of benzo(a)pyrene by S9 Mix. The ideal combined groups of benzo(a)pyrene and GE-132 were 10 $\mu{M}$ and 10mg, 20 $\mu{M}$ and 20mg, 100 $\mu{M}$ and 30 mg, respectively. Then, the revertants in antimutagenicity test, which was studied the direct action of GE-132 on the induction of revertant cells by Salmonella typhimurium TA98 and activated benzo(a)pyrene were decreased significantly in the treated groups of GE-132 than no treated groups. The number of revertants of Salmonella typhimurium TA98 were reduced with increasing amounts of GE-132. From the above results, it was found that GE-132 inactivated the mutagenic metabolites of benzo(a)pyrene without inhibition of the enzyme action in the S9 Mix, and GE132 showed antimutagenic effect which have inhibitory action of reverted gene expression.

• 한국생물공학회:학술대회논문집
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• 한국생물공학회 2003년도 생물공학의 동향(XII)
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• pp.445-449
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• 2003

As for the purpose, we first introduce an random mutation into wild-type gene to expand a mutation space, and then further recombine the mutant genes by staggered extension process PCR. As a result, we obtained the best clones 6-52 that showed a high activity and stability, from a round of error prone and staggered extension process PCR. The purified enzyme showed a similar pH stability to the wild-type enzyme and reveal a slightly high optimum pH at 12. In the optimum temperature, an identical dependency was also showed and a quite high stability in the thermal stability was obtained. Along with this, the enzyme was also stable at a reaction that supplement with a 15 % of ethanol as an additive. The addition of other solvents and surfactants did not improve the reaction and thus resulted in a similar profile to those of wild-type enzyme. The specific activity on the target compound rac-ketoprofen ethyl ester was calculated to be about 85, 000 unit, and the kinetic constants Km and Vmax were determined to be 0.2 mM and 90 mM/mg-protein/min respectively. The deduced amino acid alignment with the wild type enzyme revealed five mutations at L120P, I208V, T249A, D287H and T357A. Based on these observations, the site directed mutagenesis to delineate the mutagenic effect is under progress.

• Toxicological Research
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• 제11권1호
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• pp.31-36
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• 1995

Cyclophosphamide (CP) must be enzymatically activated by cytochrome P450(CYP)-linked mixed-function oxidation pathway to be either mutagenic or teratogenic. Influences of alterations in hepatic mixed-function oxidase acitivity and glutathione (GSH) content on the embryotoxicity of CP were studied in rat whole embryo culture system. The embryotoxicity of CP was compared using rat S-9 fraction (S-9) pretreated with chemicals inducing different CYP isozymes, acetone (ACE), Aroclor 1254 (ARO), $\beta$-naphthoflavone (NAF) and phenobarbital (PHE). When 10.5 day embryos were cultured in the immediately centrifuged rat serum for 48 hrs using general gas char{ging schedule, CP$(40{\mu}g/ml)$ with S-9 induced by either NAF or PHE increased the incidence of realformations and significantly decreased embryonic growth compared with the non-induced S-9 group. ACE or ARO induced S-9 group showed no significant difference in embryonic growth. These data suggest that PB and/or NAF inducible CYP isoenzymes are mainly involved in the activation of CP. To examine the effect of GSH on the embryotoxicity of CP, 10.5 day embryos were exposed to CP and S-9 after preincubation with 10 mM of GSH for 3 hrs. In the GSH pretreated group the growth of embryos increased significantly compared with that of the untreated group, suggesting that GSH may protect embryos in culture from some toxic effects of CP.

• Journal of Microbiology and Biotechnology
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• 제11권2호
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• pp.346-349
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• 2001

The antimutagenic activity of polysaccharides extracted from soybeans fermented with Agrocybe cylindracea (AC) or Phellinus igniarius (PI) against 2-amino-3,8-dimethylimidaxo[4,5-f]quinoxaline (MeIQx) was examined using a Salmonella/Ames test and host-mediated assay in mice. The polysaccharides from the soybeans fermented with A. Cylindracea and P. igniarius inhibited the mutagenic acitivity of the cooked food mutagen, MeIQx, by 31.2% and 35.3%, respectively. The polysaccharides also inhibited MeIQx genotoxicity in a dose-dependent manner in micel. These results suggest that the polysaccharides from soybeans fermented with A. cylindracea or P. igniarius exhibit antimutagenic properties against MeIQx in vitro and in vivo.

• 한국대기환경학회지
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• 제12권3호
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• pp.269-278
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• 1996

Airborne suspended particulates were collected by an Andersen high volume air sampler in a traffic area of Seoul from September 1990 to August 1991. Origanic matter extracted from particulates, their fractions, namely acidic, basic, neutral and carcinogenic subfractions (PAHs, nitroarenes) in neutral fractions were assayed for mutagenicity on TA98, TA100 and TA98NR deficient Salmonella strains, use of the pre-incubation method. The relative contribution to total mutanenicity of organic matters was highest in neutral fraction and was lowest in basic fraction. Among subfractions, that of neutral fraction was higher nitroarenes subfraction compared to PAHs subfraction. While the carcinogenic effect of benzo[a]pyrene was calculated as 0.96 persons/million persons based on unit risk estimates by extrapolation method, life time excess cancer risk estimate of EOM, neutral, PAH fraction based on their mutagenicity was calculated as 52, 42, 3.8 persons/million persons, respectively. These findings indicate that the mutagenic hazard of the partciculate, air organic complex mixture, may be dependent upon the mutagen composition in the particulate and interactions each of them. Therfore, health risk from air organic complex mixtures based on mutagenicity might be useful indicator for evaluation of actual risk.

• Journal of Radiation Protection and Research
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• 제27권2호
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• pp.81-87
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• 2002

The interaction of low density extremely low frequency magnetic field (ELF MF) in the frequency of hypoxanthine-guanine phosphoribosyltransferase (HPRT) mutation induced by bleomycin and on the frequency of sister chromatid exchanges (SCEs) induced by 1,2,4-benzenetriol(BT) was demonstrated. CHO cells pretreated with bleomycin or 1,2,4-benzenetriol were exposed for 24hrs to a sinusoidal 0.8mT magnetic field at 60Hz. Frequency of HPRT mutation and SCEs were determined. ELF MF exposure led to a two-fold increase of the frequency of HPRT mutation induced by bleomycin. No increase of mutation frequency was observed by ELF MF alone ELF MF also increased the frequency of SCEs induced by BT while no Increase of SCE frequencies were observed by ELF MF alone. These results suggest that low density ELF MF field would art as an enhancer rather than as an initiator of mutagenic effects in CHO cell.

• 한국환경성돌연변이발암원학회지
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• 제22권2호
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• pp.97-105
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• 2002

Nowadays a huge variety of products that aim to assist to quit smoking or reduce addictive symptoms are developed and manufactured with safety evaluation, but the safety of the most recent products of interest which do not contain tobacco and nicotine, and shape cigarettes is not evaluated and guaranteed relatively. This study was carried out to evaluate the single and repeated dose inhalation toxicity and genotoxicity of H menthol (Nicotine free-tobacco free) herbal cigarettes provided by Cigastop Ltd. in ICR mice. In this study, doses which we determined to expose to mice were 40 cigarettes for 6 hours a day to mice in single dose and 20 (high dose), 10 (middle dose) and 5 cigarettes (low dose) a day for 28 days in repeated dose inhalation toxicity, in vivo chromosome aberration test and micronucleus test. The particulate substances from H menthol herbal cigarettes also were gathered and used in the Salmonella typhimurium/microsome assay (Salmonella test; Ames test). We could find neither significant changes between control and treatment groups nor dose-response effects of test material at all except serum Ca level of female middle dose treatment group in repeated dose inhalation toxicity test. In conclusion, H menthol herbal cigarettes, when applied clinically intended dose we used, might not show any toxic and/or mutagenic effect.

• 한국환경과학회지
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• 제4권5호
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• pp.157-157
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• 1995

In order to analyze the sex-linked lethal mutagenic effects of ethyl methanesulfonate (EMS) in Drosophila melanogaster, the mutagenicities from the attached-X and Basc method have been detected. The indexes of relative lethal mutations at 2.0 and 4.0mM EMS treated group using the attached-X method were about 1.5 and 2.4 times than that of 1.0mM treated group, respectively. EMS had more pronounced effect in the sperm and spermatid stages in the induction of X-linked lethal mutations during the spermatogenesis. And, in the induction of X-linked recessive lethal mutations from the Basc method, the mutation frequency of 4.0mM EMS treated group as compared with 2.0mM was more than three fold. Two assay systems used in this study can support mutually according to experimental purposes, and the area of its application can be considerably wide.