낙동강 하구수의 변이원성에 대한 연구

윤명희,김지혜,大津隆一,민병윤 한국환경과학회 2001 한국환경과학회지 Vol.10 No.1

낙동강 하구수에서의 변이원성 물질에 의한 오염 상황을 파악하기 위하여 1998년에 각 계절별로 1회씩 총 4회에 걸쳐 낙동강 하구 10개 지점에서 blue rayon법을 이용한 Ames test를 실시하여, 3환 이상의 다환화합물에의한 변이원성을 조사하였다. 이 중 5월에 공업지역 부근인 제 1지점(하단동 부근)과 제 4지점(장림동 부근)에서 채취된 시료에서 현저한 변이원성이 관찰되었는데, 제 1지점의 시료에서는 S9 mix존재 및 비존재 하의 TA98, TA100 세포주 모두 현저한 용량 의존적 양성반응을 나타내어, 이 지점은 frame shift형의 직접 및 간접돌연변이형 원인 물질과 염기치환형 직접 및 간접돌연변이형 원인 물질에 의하여 오염되어 있음이 밝혀졌다 또한 제 4지점의 시료에서는 S9 mix존재 및 비존재 하에서 TA98세포주가 양성반응을 나타내어, 이 지점에 대한 변이원성의 돌연변이형은 fram shift형의 직접 및 간접 돌연변이임을 시사해주고 있다. 그러나 7월 이후에는 변이원성이 관찰되지 않았는데, 이는 1998년의 경제성장률의 저하에 기인된 것으로 생각되었다. 한편, 주거지역에 인접한 제 5-10지역의 시료에서는 전 조사기간에 걸쳐 변이원성이 관찰되지 않았는데, 이는 주된 오염원이 생활하수인 이들 지역의 강물이 변이원성 물질에 의해 오염되어 있지 않았음을 시사해 주고 있다. The mutagenicity of the river water of Nakdong river estuary was determined by Ames test using the blue rayon suspension method. Samples were collected from 10 sites in the estuary once in each season of 1998. The samples collected from the sites where industrial waste water discharge on May were mutagenic, but the other samples were not mutagenic. The sample collected from the site 1 located near the industrial area (Hadan-dong) were highly mutagenic in the TA98 with (+S9) and without (-S9) S9 mix as well as in the TA100 with (+S9) and without (-S9) S9 mix, suggesting that the river water of this site is polluted by direct and indirect mutagens of frame-shift type as well as direct and indirect mutagens of base-replacement type. The positive mutagenicity, although relatively low, was also detected in TA98 with (+S9) and without (-S9) S9 mix in the extract of the site 4 near the industrial area(Jangrim-dong), suggesting that the primary mutation type is frame-shift. The negative mutagenicity from July to December at the sites (1-4) near the industrial area seems to be affected by the low economic growth rate in 1998 in Korea. On the other hand, the negative mutagenicity in all extracts collected from the sites 5-10 near the residential area where living sewage discharge, suggests that the river water was not polluted by mutagens.

뇨 돌연변이 유발성 시험법 정립

장미,신한재,박철훈,손형옥,현학철,Jang, Mi,Shin, Han-Jae,Park, Chul-Hoon,Sohn, Hyung-Ok,Hyun, Hak-Chul 한국연초학회 2015 한국연초학회지 Vol.37 No.1

Urinary mutagenicity is widely recognized as a useful biomarker for the assessment of mutagen exposure level in human. In this study, we optimized the several parameters affecting the activity of Urinary mutagenicity using highly sensitive mutation test(microsuspension assay) instead of the conventional Ames test. First of all, we chose YG1024 as a highly sensitive strain from three str ains of Salmonella typhimurium(TA98, TA100, YG1024) using r epr esentative mutation substances, such as Benzo[a]pyrene, 2-Aminonaphthalene, 2-amino-3-methyl-9H-pyrido[2,3-b]indole($MeA{\alpha}C$) and cigarette total particulate matter(TPM). And we established the several kinds of test conditions such as number of bacter ia, concentr ation of metabolic activation system and incubation time for the most sensitive reaction. Also, we optimized efficient pre-treatment method using commercial C18 column. As a r esults, this method was shown a aver age of 94 % recovery value and 13 % relative standard deviation. When we compared the Urinary mutagenicity between several participants, we confirmed that compar ative measurements were possible for different levels of urine mutagenicity. In conclusion, the optimized highly sensitive mutation test to measure the Urinary mutagenicity may be useful in biological monitoring of mutagen exposure level.

안태음의 변이원성 및 간독성에 관한 연구

이동녕,문진영,오규석,이태균,최미정,이동목,남경수,Lee, Dong-Nyung,Moon, Jin-Young,Oh, Gue-Suc,Lee, Tae-Kyun,Choi, Mi-Jung,Lee, Dong-Mok,Nam, Kyung-Soo 한국생약학회 1997 생약학회지 Vol.28 No.3

Antaeum(ATE) has been used as a prescription for threatened abortion, associated with pregnancy in traditional medicine. Because gravida could be administered ATE for a long period, its administration might cause a harmful effect on fetus and gravida during the pregnancy. This study aimed to determine whether exposure to ATE caused mutagenicity or hepatotoxicity during the pregnant period. For mutagenicity test of ATE, Salmonella typhimurium and Bacillus subtilis were used as indications for DNA damage. In the Ames test, Samonella typhimurium TA98 and TA100 were used for mutagenicity testing, and the number of histidine revertants was measured. In Rec-assay, Bacillus subtilis H $17(Rec^+)$ and $M-45(Rec^-)$ strains were used to clarify the DNA damage property. In the SOS umu test, Salmonella typhimurium TA15335 containing plasmid pSK1002 was used as a tester strain, and we monitored the levels of umu operon expression by measuring the ${\beta}-galactosidase$ activity. From the tested results, ATE did not show DNA damage and mutagenicity. On the other hand, hepatotoxicity of ATE to female ICR mice was monitored by the measurements of s-GOT, s-GPT and LDH activities after oral feeding for 15 days. ATE did not show significant change of s-GOT, s-GPT and LDH activities in mice sera.

Mutagenicity Assessment of Drinking Water in Combination with Flavored Black Tea Bags: a Cross Sectional Study in Tehran

Alebouyeh, Farzaneh,Bidgoli, Sepideh Arbabi,Ziarati, Parisa,Heshmati, Masoomeh,Qomi, Mahnaz Asian Pacific Journal of Cancer Prevention 2015 Asian Pacific journal of cancer prevention Vol.16 No.17

Diseases related to water impurities may present as major public health burdens. The present study aimed to assess the mutagenicity of drinking water from different zones of Tehran, and evaluate possible health risks through making tea with tea bags, by Ames mutagenicity test using TA 100, TA 98 and YG1029 strains. For this purpose, 450 water samples were collected over the period of July to December 2014 from 5 different zones of Tehran. Except for one sample, no mutagenic potential was detected during these two seasons and the MI scores were almost normal (${\leq}1-1.6$) in TA 100, TA 98 and YG1029 strains. Although no mutagenic effects were considered in TA 98 and TA 100 in the test samples of our three evaluated tea bag brands, one sample from a local company showed mutagenic effects in the YG1029 strain (MI=1.7-1.9 and 2) after prolonged (10-15 min.) steeping. Despite the mild mutagenic effect discovered for one of the brand, this cross sectional study showed relative safety of water samples and black tea bags in Tehran. According to the sensitivity of YG1029 to the mutagenic potential of water and black tea, even without metabolic activation by s9 fraction, this metabolizer strain could be considered as sensitive and applicable to food samples for quantitative analysis of mutagens.

Computational Study of Mutagen X

Cho, Seung-Joo Korean Chemical Society 2003 Bulletin of the Korean Chemical Society Vol.24 No.6

Mutagen X (MX), 3-chloro-4-(dichloromethyl)-5-hydroxy-2(5H)-furanone is one of the most potent directing acting mutagen ever tested in SAL TA100 assay. Although MX analogues have been synthesized, tested for mutagenicity and modeled by structure-activity relationship (SAR) methods, the mechanism of interaction of these compounds with DNA to produce their remarkable mutagenic potency remains unresolved. MX exists as an equilibrium mixture of both ring and open form in water. This equilibrium is very fast for Ames test. Because the mixture is not separable by experimental methods, it is not clear which one is really responsible for the observed mutagenicity. There have been many debates that which one is really responsible for the observed mutagenicity. We used ab initio methods for the MX analogues. It seems both ring and open form could react with DNA bases as electrophiles. However, every open form has consistently lower LUMO energy than corresponding ring form. It is reasonable to assume that the major reaction will go through via open form for MX analogues. This suggest that the open form is more likely really mutagenic.

Development of Fecal Microbial Enzyme Mix for Mutagenicity Assay of Natural Products

( Hee Kyung Yeo ),( Yang Jin Hyun ),( Se Eun Jang ),( Myung Joo Han ),( Yong Sup Lee ),( Dong Hyun Kim ) 한국미생물 · 생명공학회 2012 Journal of microbiology and biotechnology Vol.22 No.6

Orally administered herbal glycosides are metabolized to their hydrophobic compounds by intestinal microflora in the intestine of animals and human, not liver enzymes, and absorbed from the intestine to the blood. Of these metabolites, some, such as quercetin and kaempherol, are mutagenic. The fecal bacterial enzyme fraction (fecalase) of human or animals has been used for measuring the mutagenicity of dietary glycosides. However, the fecalase activity between individuals is significantly different and its preparation is laborious and odious. Therefore, we developed a fecal microbial enzyme mix (FM) usable in the Ames test to remediate the fluctuated reaction system activating natural glycosides to mutagens. We selected, cultured, and mixed 4 bacteria highly producing glycosidase activities based on a cell-free extract of feces (fecalase) from 100 healthy Korean volunteers. When the mutagenicities of rutin and methanol extract of the flos of Sophora japonica L. (SFME), of which the major constituent is rutin, towards Salmonella typhimurium strains TA 98, 100, 102, 1,535, and 1,537 were tested using FM and/or S9 mix, these agents were potently mutagenic. These mutagenicities using FM were not significantly different compared with those using Korean fecalase. SFME and rutin were potently mutagenic in the test when these were treated with fecalase or FM in the presence of S9 mix, followed by those treated with S9 mix alone and those with fecalase or FM. Freeze-dried FM was more stable in storage than fecalase. Based on these findings, FM could be usable instead of human fecalase in the Ames test.

Evaluation of New Direct Dyes for Mutagenicity Using the Salmonella Mammalian Mutagenicity Assay

Bae Jin-Seok,Freeman Harold S. The Korean Fiber Society 2005 Fibers and polymers Vol.6 No.4

A series of new direct dyes based on benzidine congeners, 2,2'-dimethyl-5,5'-dipropoxybenzidine and 5,5'-dipropoxybenzidine, were evaluated for mutagenicity in Salmonella typhimurium strains TA98 and TA100. All of the dyes examined were judged to be non-mutagenic with and without metabolic activation while toxicity was seen in some dyes at high doses. The study also suggested that the standard Salmonella mutagenicity plate-incorporated assay was an excellent method for evaluation of direct dyes for mutagenicity.

쑥 추출물의 항돌연변이 활성효과

이성,권동진,유진영,정동효 한국미생물생명공학회 ( 구 한국산업미생물학회 ) 1996 한국미생물·생명공학회지 Vol.24 No.1

한국산 쑥으로부터 얻어진 ethanol 추출물과 각 용매획분에 대하여 in vitro로서 Salmonella typhimurium reversion assay 시험을 하여 자체 변이원성 유무와 4가지 양성 변이원 물질(3㎍/plate 2NF, 2㎍/plate AFB_1, 1㎍/plate Trp-P-1 그리고 1㎍/plate Trp-P-2)에 대한 항돌연변이 활성에 대한 쑥의영향을 고찰하였다. 쑥의 ethanol 추출물은 2NF, AFB_1, Trp-P-1 그리고 Trp-P-2 등으로 유도한 돌연변이 시험(Ames test)에서 변이원성을 보여주지 않았다. 돌연변이 억제시험에서는 frameshift type인 Sal. typhimurium TA 98에 대하여 직접 변이원인 2-NF에서는 19.7~22.9%, 간접 변이원인 AFB_1, Trp-P-1과 Trp-P-2에서는 각각 47.9~61.2%, 64.1~70.7% 그리고 67.4~78.7%의 억제효과를 나타내었다. 또한 쑥의 ethanol 추출물을 n-hexane, chloroform, ethyl acetate 그리고 water로 각각 분획하여 직접변이 원인 2-NF와 간접변이원인 AFB_1을 사용하여 변이원성과 항돌연변이원성 시험을 하였다. 돌연변이 시험에서 변이원성은 4가지 획분에서 모두 나타나지 않았다. 직접 변이원인 2-NF로 유도된 돌연변이에 대하여 n-hexane 획분은 9.4~33.7%, chloroform 획분은 21.6~26.5% 그리고 ethytl acetate 획분은 2.4~21.9%를 나타내었다. 간접 변이원인 AFB_1로 유도된 돌연변이에 대하여 n-hexane 획분, chloroform 획분 그리고 ethytl acetate 획분들은 각각 63.0~80.0%, 77.5~82.1%, 68.5~83.1%로 높은 억제 효과를 보여주었다. Mugwort has been know as a traditional substitutive foodstuff and as showing a physiologically beneficial function to a human being. Therefore, effect of mugwort extract in terms of mutagenicity and desmutagenicity was investigated to verify its function. Ethanol extract from mugwort did not exhibit any mutagenicity. On the contrary, inhibitory effects of the ethanol extract were observed on mutagenicity induced by aflatoxin B_1(AFB_1), 3-amino-1,4-dimethyl-5H-pyrido[4,3-b]indole(Trp-P-1), 3-amino-1-methyl-5H-pyrido[4,3]indole(Trp-P-2) and 2-nitroflourene(2NF) using Salmonella typhimurium reversion assay. On direct-acting mutagen(2NF, 3 ㎍/plate), ethanol extract showed a slight inhibitory effect of 19.7%~22.9%, however on indirect-acting mutagen such as AFB1(2 ㎍/plate), Trp-P-1(1 ㎍/plate) and Trp-P-2(1 ㎍/plate), we observed higher inhibitory effect of 47.9~61.2%, 64.1~70.7%, 67.4~78.7%, respectively. Step-wise fractionation of the ethanol extract was done by using hexane, chlorofractions showed high inhibition of 63.0~80.0%, 77.5~82.1%, and 68.5~83.1%, respectively on the mutagenicity of AFB_1 in Sal. typhimurium TA98. Consequently, these results indicated that mugwort extract contains some compound(s) which may show desmutagenicity.

Evaluation of New Metallized Direct Dyes for Mutagenicity Using the Salmonella Mammalian Mutagenicity Assay

Rae Jin-Seok,Freeman Harold S. The Korean Fiber Society 2005 Fibers and polymers Vol.6 No.3

A series of new metallized direct dyes based on benzidine congeners, 2,2'-dimethyl-5,5'-dipropoxybenzidine and 5,5'-dipropoxybenzidine, were evaluated for mutagenicity in Salmonella typhimurium strains TA98 and TA 100. All of the dyes examined were judged to be non-mutagenic with and without metabolic activation while toxicity was seen in some dyes at high doses. The study also suggested that the standard Salmonella mutagenicity plate-incorporated assay was an excellent method for evaluation of dyes for mutagenicity.

교애사물탕의 변이원성 및 간독성에 관한 연구

우덕안,홍희탁,문진영,이태균,김철호,김준기,최미정,남경수 한국독성학회 1997 Toxicological Research Vol.13 No.3

Kyoaesamultang(KAT) has been used as an important prescription for various diseases including threatened abortion, associated with pregnancy in traditional medicine. In oder to identify the safety of KAT, this study was designed to determine mutagenicity and hepato-toxicity. In Rec-assay, Bacillus subtills H-17($Rec{^+}$) and M-45($Rec{^-}$) strains were used to clarify the DNA damage property. In Ames test, Salmonella typhimurium TA98 and TA100 were used for mutagenicity testing. In SOS umu test, Salmonella typhimurium TA1535 containing plasmid pSK1002 was used as a tester strain, and the levels of umu operon expression were monitored by measuring the $\beta$-galactosidase activity. From tested results, KAT did not show DNA damage and mutagenicity. On the other hand, hepato-toxicity of KAT to female ICR mice was monitored by the measurements of s-GOT, s-GPT and LDH activities after oral feeding for 15days. KAT showed 34% increase of s-GOT and s-GPT activities, also exhibited 35% increase of LDH activity in mice sera.