• Title/Summary/Keyword: Rec assay

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Mutagenicity and Hepato-Toxicity of Kyoaesamultang (교애사물탕의 변이원성 및 간독성에 관한 연구)

  • 우덕안;홍희탁;문진영;이태균;김철호;김준기;최미정;남경수
    • Toxicological Research
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    • v.13 no.3
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    • pp.197-202
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    • 1997
  • 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.

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Antimutagenic Effect of Korean Mistletoe Extracts (겨우살이 추출물의 항돌연변이 효과)

  • 함승시;강신태;최근표;박원봉;이득식
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.27 no.2
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    • pp.359-365
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    • 1998
  • This study was carried out to investigate mutagenecity and antimutagenic effects from crude extract, heating extract and alcohol extract of Korean mistletoe(Viscum album L.) on the bacterial short-term tests, such as Ames test, spore rec-assay, SOS spot test and SOS chromotest by using several kinds of mutagens. In the Ames test, each extract did not show any mutagenesis, but each extract showed inhibitory effects of 80∼95% and 70∼94% against mutagenesis induced by 3-amino-1, 4-dimethyl-5H-pyrido[4,3-b] indole(Trp-P-1) and 2-aminofluorene(2-AF) in Salmonella typhimurium TA98, respectively. In th spore rec-assay, mistletoe ectracts showed antimutagenic effect with inhibiton zone in the range of 5∼11mm against mutagenicity induced by mitomycin C(MMC, 18mm) and N-methyl-N'-nitro-N-nitrosoguanidne (MNNG, 24mm), respectively. The heating and alcohol extracts in the SOS chromotest showed 96% and 70% inhibition against benzo-α-pyrene[B(α)P] and Trp-P-1 induced mutagenesis, respectively.

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Cytotoxicological and Pathological Studies of 6-Paradol, a Pungent Principle of Ginger (생강성분 6-Paradol의 세포 독성 및 병리학적 연구)

  • Kim, Ok-Hee;Yoo, Eun-Sook;Jung, In-Kyung;Lee, Sang-Sup
    • Environmental Mutagens and Carcinogens
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    • v.18 no.1
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    • pp.32-36
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    • 1998
  • It is previously reported that 6-paradol can induce prolonged analgesia in experimental animals. In order to investigate the mutagenicity of 6-paradol, Ames Samonella/microsome plate assay was carried out with Salmonella typhimurium strains, TA 98, TA 100, TA 1535 and TA 1538, 6-Paradol was nonmutagenic in Salmonella typhimurium with and without rat liver microsomal activation. The rec assay with Bacillus substilis strains H 17 $rec^+$ and M 45 $rec^+$ was carried out ot test 6-paradol and other compounds (1-3 mg/disc) for DNA damaging activity, 6-Paradol was also nonmutagenic in DNA damaging activity. The relative size of the inhibition zone for 6-paradol was smaller than that of capsaicin. We have also determined the pathological effects of this compound on the various tissues of rats after administrating(i.p.) with increasing doses of 4, 8, 12, 16 mg/kg at 2 hour intervals and found no significant changes in terms of histology.

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Desmutagenicity of Enzymatically Browned Substances Obtained from the Reaction of Prunus salicina (Red) Enzyme and Polyphenols (재래종 적색자두(Prunus salicina) 효소갈변반응 생성물의 돌연변이 억제작용)

  • Ham, Seung-Shi;Hong, Eun-Hee;Omura, Hirohisa
    • Korean Journal of Food Science and Technology
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    • v.19 no.3
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    • pp.212-219
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    • 1987
  • The rec-assay on Bacillus subtilis strains H17$({Rec}^+)$ and M45$({Rec}^-)$, the Ames test with modification of preincubation on Salmonella typhimurium TA98 and TA100 and DNA-breaking test on double strand calfthymus DNA were carried out using enzymatically browned substances obtained from the reaction of Prunus salicina (Red) enzyme and polyphenols. The spore rec-assay of enzymatic browning reaction products of pyrogallol, hydroxyhydroquinone. 3,4-dihydrohyoluene and chlorogenic acid showed non-mutagenic activity The spore rec-assay showed a little influence of ${Zn}^{2+}$ and ${Ni}^{2+}$ on the action of four kinds of enzymatic browning reaction products. The enzymatic browning reaction products of polyphenols did not show DNAbreaking activity. ${Cu}^{2+}$ of various metal ions influenced on DNA-breaking of enzymatic browning reaction products of pyrogallol. However, enzymatic browning reaction products of chlorogenic acid inhibited on DNA-breaking activity. Four kinds of enzymatic browning reaction products showed non-mutagenic activity on Salmonella typhimurium TA98 and TA100 with S-9 mix. In the mutagenicity on Salmonella typhimurium TA98 and TA100 with S-9 mix in the presence of benzo$({\alpha})$pyrene which is the carcinogenic substances, four kinds of enzymatic browning reaction products showed desmutagenic activity.

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Inhibitory Effect of Persimmon Leaves on the Mutagenicity in Spore Rec Assay and on the Growth of Human Cancer Cells (감잎의 용매별 추출물의 돌연변이 유발 억제 및 암세포 증식억제 효과)

  • 문숙희
    • The Korean Journal of Food And Nutrition
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    • v.15 no.1
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    • pp.23-28
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    • 2002
  • The inhibitory effects of persimmon leaves on th e mutagenicity in spore rec assay and on the growth of HT-29 human colon cancer cells and AZ-521 human gastric cancer cells were studied. Methanol extract of persimmon leaves inhibited the mutagenicity induced fly N-methyl- N'-nitro-N-nitrosoguanidine(MNNG) in spore rec assay. The hexane, chloroform and ethylacetate fraction from the methanol extract exhibited strong antimutagenicity against MNNG in spore rec assay The methanol extract of persimmon leaves also revealed the inhibitory effects on the growth of HT-29 human colon cancer cells and AZ-521 human gastric cancer cells. Among the solvent extracted fraction from the methanol extract, the chloroform fraction was most effective and inhibited the growth of HT-29 and AZ-521 cells by 100 percent.

Inhibitory Effect of Main Pine Needle Extracts on the Chemically Induced Mutagenicity (주요 솔잎 추출물의 돌연변이 억제효과)

  • Kim, Eun-Jeong;Jung, Sung-Won;Choi, Keun-Pyo;Ham, Seung-Shi;Kang, Ha-Young
    • Korean Journal of Food Science and Technology
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    • v.30 no.2
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    • pp.450-455
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    • 1998
  • Pine has been known as a traditional medicinal plant and as showing a physically beneficial function to a human being. Therefore, this study was performed to investigate the physiological activities of main pine neddles. Ethanol extracts from pinus needles did net exhibit any mutagenicity. On the contrary, inhibitory effects of ethanol extract were observed on mutagenicity induced by N-methyl-N'-nitro-N-nitrosoguanidine(MNNG), 4-nitroquinoline-1-oxide (4NQO), 3-amino-1,4-dimethyl-5H-pyrido-(4,3-b)indol (Trp-P-1) and benzo(a)pyrene $(B({\alpha})P)$ using Salmonella typhimurium reversion assay. On direct-acting mutagen (MNNG, 4NQO) and indirect-acting mutagen (Trp-P-1, $(B({\alpha})P)$, we observed higher inhibitory effect. Stepwise fractionation of the ethanol extract was done by using ether, chloroform, ethyl acetate, butanol and water to obtain effective fraction. Among them, water fractions $(100\;{\mu}g/plate)$ of Pinus thunbergii, Pinus rigida, Pinus densiflora and Pinus koraiensis showed high inhibition of 91.65%, 94.7%, 84.22% and 79.02%, respectively, on the mutagenicity of MNNG in Salmonella typhimurium TA100.

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EXAMINATION OF TYR-264 FOR ATPase ACTIVE SITE IN E.coli RecA PROTEIN BY SITE-DIRECTED MUTAGENESIS

  • Kwon, Yong-Kook;Bae, Jun-Seong;Hahn, Tae-Ryong
    • Journal of Photoscience
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    • v.2 no.1
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    • pp.27-29
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    • 1995
  • Site directed mutagenesis has been introduced to determine active site(s) and molecular structure of E. coli RecA protein. Recombinant DNAs were constructed by point mutation of Tyr-264 to Phe which assumed active site for binding and hydrolysis of ATP. RecA proteins were purified from recombinants containing wild type and mutant genes and analyzed for ATPase activity assay. Result suggests that Tyr-264 is involved in ATP binding rather than ATP hydrolysis.

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Genotoxicity Assay Using Chromosomally-Integrated Bacterial recA::Lux

  • Min, Ji-Ho;Gu, Man-Bock
    • Journal of Microbiology and Biotechnology
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    • v.13 no.1
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    • pp.99-103
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    • 2003
  • An Escherichia coli strain containing the recA promoter that fused to the luxCDABE operon originating from Photorhabdus luminescens was shown to respond sensitively to genotoxic stresses. Two different recombinant bacteria, one (DPDI 657) harboring a plasmid with the recA promoter that fused to the luxCDABE operon, and the other (DPD1710) containing a chromosomally-integrated recA promoter that fused with luxCDABE, were compared and it was found that the sensitivity of 'the two strains was significantly different in terms of their bioluminescent level, response time, and the minimum detectable concentration of a chemical causing DNA damaging stress. DPDI 710, with a chromosomally-integrated single copy, generally led to lower basal luminescence levels, faster responses, increased response ratios, and an enhanced sensitivity to mutagens, when compared to DPD 1657 with a multi-copy plasmid.

Isolation and Characterization of Lactic Acid Bacteria Producing Antimutagenic Substance from Korean Dongchimi (동치미로부터 항돌연변이 물질을 생산하는 유산균의 분리 및 특성)

  • 주길재;이창호;우철주
    • Journal of Life Science
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    • v.11 no.5
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    • pp.432-438
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    • 2001
  • Various lactic acid bacteria were isolated from Korean Dongchimi (whole radish Kimichi with added water) in order to study their antimutagenic activity. Ames test using Salmonella enterica serovar typhimurium TA98 and TA100 showed the strain DLAB19 to have the highest antimutagenic activity among the 300 isolated strains against MNNG(N-methyl-N-nitro-N-nitrosoguanidine), NPD (4-nitro-O-phenylenediamine), 4-NQO(4-nitroquinoline-1-oxide) and AFB$_{1}$(aflatoxin B$_{1}$). The strain was identified as Leuconostoc mesenteroides subsp. cremoris according to the Bergeys Mannual Systematic Bscteriology based on its morphological, cultural, physiological characteristics and biological system Antimutagenic activity of Leu. mesenteroides subsp. cremoris DLAB19 was found in the culture supernatant suggesting the bacterium secretes, the antimutagenic substance in the media. The antimutagenic activity of Leu. mesenteroides subsp. cremoris DLAB19 was reconfirmed by the spore-rec assay using spores of Bacillus subtilis H17 (Rec$^{+}$) and M45 (Rec$^{[-10]}$ ).).

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Inhibitory Effect of Rice Extract on the Chemically Induced Mutagenesis (쌀 추출물의 돌연변이 억제효과)

  • Chun, Hyang-Sook;Kim, In-Ho;Kim, Young-Jin;Kim, Kil-Hwan
    • Korean Journal of Food Science and Technology
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    • v.26 no.2
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    • pp.188-194
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    • 1994
  • The inhibitory effects of rice extract on mutagenicity induced by 3-amino-1,4-dimethyl-5H-pyrido [4,3-b]indole(Trp-P-1), 3-amino-1-methyl-5H-pyrido [4,3-b]indole(Trp-P-2), sodium azide(SA), 2-nitrofluorene(2NF), mitomycin C(MMC), aflatoxin $B_1(AFB_1)$ and 4-nitroquinoline oxide(4-NQO) were investigated using Salmonella typhimurium reversion assay, SOS chromotest and spore rec-assay. In Salmonella typhimurium reversion assay, methanol extract from brown rice (Illpumbyeo, Japonica variety) showed the highest inhibitory effect among other extracting solvent including hexane, chloroform and water. Methanol extract showed stronger inhibitory effect, above 85%, on indirect-acting mutagens(Trp-P-1, Trp-P-2 and $AFB_1$) than those on direct-acting mutagens(4-NQO, 2NF). In SOS chromotest, methanol extracts showed $77.6{\sim}88.9%$ effects on SOS function induced by Trp-P-1, Trp-P-2, $AFB_1$ and 4-NQO. In spore rec-assay, methanol extracts inhibited the mutagenicity induced by $AFB_1$ and MMC. As the concentration of methanol extract increased, inhibitory effect on mutagenicity increased but reached at steady state as inhibition rate of 90% when the concentration was above 5 mg/plate. In inhibitory effects of methanol extracts by various rice varieties, all of 11 varieties turned out to have inhibitory effect on mutagenicity. There was no significant difference (p>0.05) in inhibitory effect of methanol extracts between brown and white rice against Trp-P-1, but showed difference (p<0.05) against 4-NQO.

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