• Title/Summary/Keyword: bacterial reverse mutation assay

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Genotoxicity on $21{\alpha}-and\;{\beta}-methylmelianodiol$, a Component of Poncirus trifoliata, in Bacterial and Mammalian Cells

  • Ryu, Jae-Chun;Kim, Youn-Jung;Kim, Mi-Soon;Kim, Min-Ji;Sarma, Sailendra Nath;Lee, Seung-Ho
    • Molecular & Cellular Toxicology
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    • v.1 no.3
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    • pp.172-178
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    • 2005
  • [ $21{\alpha}$ ]- and ${\beta}$-Methylmelianodiol were isolated as the inhibitor of IL-5 bioactivity from Poncirus tripoliata. To develope as an anti-septic drug, the genotoxicity of $21{\alpha}\;-and\;{\beta}-methylmelianodiol$ was subjected to high throughput toxicity screening (HTTS) because they revealed strong IL-5 inhibitory activity and limitation of quantity. Mouse lymphoma thymidine kinase ($tk^{+/-}$) gene assay (MOLY), single cell gel electrophoresis (Comet) 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 $21{\alpha}-methylmelianodiol$ was determined the concentration of $25.51\;{\mu}g/mL\;and\;21.99\;{\mu}g/mL$ with and without S-9, respectively. Also $21{\beta}-methylmelianodiol$ was determined the concentration of $24.15\;{\mu}g/mL\;and\;\;22.46\;{\mu}g/mL$ with and without S-9, respectively. In the comet assay, DNA damage was not observed both $21{\alpha}-methylmelianodiol\;and\;21{\beta}-methylmelianodiol$ in mouse lymphoma cell line. Also, the mutant frequencies in the treated cultures were similar to the vehicle controls, and none of $21{\alpha}\;-and\;{\beta}-methylmelianodiol$ with and without S-9 doses induced a mutant frequency over. twice the background. It is suggests that $21{\alpha}\;-and\;{\beta}-methylmelianodiol$ are non-mutagenic in MOLY assay. The results of this battery of assays indicate that $21{\alpha}\;-and\;{\beta}-methylmelianodiol$ have no genotoxic potential in bacterial or mammalian cell systems. Therefore, we suggest that $21{\alpha}\;-and\;{\beta}-methylmelianodiol$, as the optimal candidates with both no genotoxic potential and IL-5 inhibitory effects must be chosen.

Genotoxicity Assessment of Erythritol by Using Short-term Assay

  • Chung, Young-Shin;Lee, Michael
    • Toxicological Research
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    • v.29 no.4
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    • pp.249-255
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    • 2013
  • Erythritol is a sugar alcohol that is widely used as a natural sugar substitute. Thus, the safety of its usage is very important. In the present study, short-term genotoxicity assays were conducted to evaluate the potential genotoxic effects of erythritol. According to the OECD test guidelines, the maximum test dose was 5,000 ${\mu}g$/plate in bacterial reverse mutation tests, 5,000 ${\mu}g/ml$ in cell-based assays, and 5,000 mg/kg for in vivo testing. An Ames test did not reveal any positive results. No clastogenicity was observed in a chromosomal aberration test with CHL cells or an in vitro micronucleus test with L5178Y $tk^{+/-}$ cells. Erythritol induced a marginal increase of DNA damage at two high doses by 24 hr of exposure in a comet assay using L5178Y $tk^{+/-}$ cells. Additionally, in vivo micronucleus tests clearly demonstrated that oral administration of erythritol did not induce micronuclei formation of the bone marrow cells of male ICR mice. Taken together, our results indicate that erythritol is not mutagenic to bacterial cells and does not cause chromosomal damage in mammalian cells either in vitro or in vivo.

Genotoxicity Study of Sophoricoside, a Constituent of Sophora japonica, in Bacterial and Mammalian Cell System

  • Kim, Youn-Jung;Park, Hyo-Joung;Kim, Young-Soo;Kim, Mi-Kyung;Lee, Seung-Ho;Jung, Sang-Hun;Ryu, Jae-Chun
    • Environmental Mutagens and Carcinogens
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    • v.21 no.2
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    • pp.99-105
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    • 2001
  • Sophoricoside was isolated as the inhibitor of IL-5 bioactivity from Sophora japonica (Leguminosae). It has been reported to has an anti-inflammatory effect on rat paw edema model. To develope as an anti-allergic drug, genotoxicity of sophoricoside was investigated in bacterial and mammalian cell system such as Ames bacterial reversion test, chromosomal aberration assay and single cell gel electrophoresis (Comet) assay. As results, in the range of 1,250~40 $\mu\textrm{g}$/plate sophoricoside concentrations was not shown significant mutagenic effects in Salmonella typhimurium TA 98, TA 100, TA 1535 and TA 1537 strains in Ames test. The 80% cell growth inhibition concentration (IC/SUB 80/) of sophoricoside was determined as above 5,000 $\mu\textrm{g}$/$m\ell$ in Chinese hamster lung (CHL) fibroblast cell and L5178Y mouse lymphoma cell line for the chromosomal aberration and comet assay, respectively. Sophoricoside was not induced chromosomal aberration in CHL fibroblast cell at concentrations of 700, 350 and 175 $\mu\textrm{g}$/$m\ell$ or 600, 300 and 150 $\mu\textrm{g}$/$m\ell$ in the absence or presence of S-9 metabolic activation system, respectively. Also, in the comet assay, the induction of DNA damage was not observed in L5178Y mouse lymphoma cell line both in the absence or presence of S-9 metabolic activation system. From these results, no genotoxic effects of sophoricoside were observed in bacterial and mammalian cell systems used in these experiments.

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Safety Evaluation of Korean Mistletoe Extract (한국산 겨우살이 추출물의 안전성 평가)

  • Kim, Inbo;Jeong, Ju-Seong;Yoon, Taek Joon;Kim, Jong Bae
    • The Korean Journal of Food And Nutrition
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    • v.26 no.3
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    • pp.383-390
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    • 2013
  • Mistlero C was shown to be non-genotoxic in a series of genotoxicity tests, including a bacterial reverse mutation test and a combined in vivo mammalian erythrocyte micronucleus test. In a bacterial reverse mutation assay, no significant increases in the number of revertant colonies, compared to the negative control, was detected in $5,000{\mu}g/plate$ of Mistlero C. In addition, with Mistlero C, no changes were shown in the number of micronucleated polychromatic erythrocytes (MNPCE) among 2,000 polychromatic erythrocytes compared to the negative control. Mistlero C was administered orally in rats to investigate acute toxicity. The $LD_{50}$ values in rats were above 2,000 mg/kg. In a repeated dose, 13-week, oral toxicity study conducted in rats, no compound-related adverse effects were shown at doses of Mistlero C of up to 1,000 mg/kg body weight/day. The results of these studies support the safe use of Mistlero C in food for human consumption.

Genotoxicity Study of Immature Green Persimmon Extract (풋감 주정 추출물의 유전독성 연구)

  • Ham, Young-Min;Yoon, Seon-A;Hyun, Ho Bong;Go, Boram;Jung, Yong-Hwan;Oh, Dae-Ju;Yoon, Weon-Jong
    • Journal of Food Hygiene and Safety
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    • v.35 no.6
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    • pp.567-573
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    • 2020
  • The persimmon is commonly cultivated in temperate regions of the world, including China, Korea, Japan, Brazil, Turkey, and Italy. In some Asian cultures, consumers are aware of the health claims related to the persimmon and its functional ingredients. The rich phytochemistry of the persimmon has opened new avenues of research on diet-based regimens designed to cure various ailments. This study was conducted to identify the genotoxicity of immature green persimmon (Diospyros kaki THUNB.) extract (DKA). The bacterial reverse mutation assay, the chromosomal aberration assay, and the mammalian micronucleus test were performed to determine the DKA genotoxicity. The result of the bacterial reverse mutation assay revealed that the DKA did not induce mutagenicity in Salmonella typhimurium TA98, TA100, TA1535, TA1537 and Escherichia coli WP2uvrA with or without metabolic activation of S9 mixture. The oral administration of DKA also caused no significant increase in the number of micronucleated polychromatic erythrocytes or in the mean ratio of polychromatic to total erythrocytes. In addition, DKA did not cause a significant chromosome aberration on CHL cells in the presence or absence of S9 activation. In conclusion, DKA could be considered as a reliable and safe functional food since no toxicity was found under the condition of this study.

Single-Dose Oral Toxicity in Rat and Bacterial Reverse Mutation Assay of Psoralea corylifolia L. Extracts (파고지 추출물의 렛트에 대한 단회 경구 투여 독성 및 복귀돌연변이능 평가)

  • Kim, Sun-A;Lim, Sun-Hye;Ahn, Ji-Yun;Kim, Sung-Ran;Ha, Tae-Youl
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.36 no.8
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    • pp.960-964
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    • 2007
  • This study was performed to examine the toxicity of Psoralea corylifolia L. by the single-dose oral toxicity tests in rat and bacterial reverse mutation assay. In single-dose oral toxicity tests, 5 mL ethanol extract of P. corylifolia L. were directly injected into 10 rats (5 males and 5 females) at a dosage of 2 g/kg. Death practice was not detected during breeding periods (14 days), and $LD_{50}$ was calculated over 2 g/kg. No difference were observed with control group in the growth rate and histological observations. In bacterial reverse mutation assay, his(-) Salmonella Typhimurium TA98, TA100, TA1535, TA1537 and trp(-) Escherichia coli WP2uvrA (pKM101) were used for assessing the toxicity of ethanol extracts of P. corylifolia L.. No significant difference in formation of the colonies and no dose-dependent increase was observed regardless of the addition of S9 mix. The results showed that ethanol extracts of P. corylifolia L. did not have single-dose oral toxicity and mutagenic toxicity.

Genotoxicological Safety Evaluation of Crude Antifungal Compounds Produced by Bacillus subtilis SN7 (Bacillus subtilis SN7이 생성한 조항균 물질의 유전독성학적 안정성평가)

  • Chang, Hae-Choon;Koh, Sang-Bum;Lee, Jae-Joon
    • The Korean Journal of Community Living Science
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    • v.28 no.1
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    • pp.131-141
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    • 2017
  • This study was carried out to perform genotoxicological safety evaluation of crude antifungal compounds produced by Bacillus subtilis SN7 (B. subtilis SN7) isolated from meju. Bacterial reverse mutation assay with Salmonella typhimurium TA98, TA100, TA1535, and TA1537 or Escherichia coli WP2uvrA in the presence and absence of the S9 metabolic activation system was carried out, and the crude antifungal compounds produced by B. subtilis SN7 showed no significant increase in the number of revertant colonies. In the chromosomal aberration tests using Chinese hamster lung (CHL) cells, sample treatment groups showed no increase in the frequency of chromosome aberrations compared to the negative control group. Furthermore, in the micronucleus formation test, the crude antifungal compounds showed no significance increase in the frequency of polychromatic erythrocytes with micronuclei. These results suggest that the crude antifungal compounds produced by B. subtilis SN7 isolated from meju showed no harmful genotoxic effects.

The First Report to Evaluate Safety of Cyanobacterium Leptolyngbya sp. KIOST-1 for Use as a Food Ingredient: Oral Acute Toxicity and Genotoxicity Study

  • Lee, Youngdeuk;Kim, Taeho;Lee, Won-Kyu;Ryu, Yong-Kyun;Kim, Ji Hyung;Jeong, Younsik;Park, Areumi;Lee, Yeon-Ji;Oh, Chulhong;Kang, Do-Hyung
    • Journal of Microbiology and Biotechnology
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    • v.31 no.2
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    • pp.290-297
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    • 2021
  • Leptolyngbya sp. KIOST-1 (LK1) is a newly isolated cyanobacterium that shows no obvious cytotoxicity and contains high protein content for both human and animal diets. However, only limited information is available on its toxic effects. The purpose of this study was to validate the safety of LK1 powder. Following Organisation for Economic Co-operation and Development (OECD) guidelines, a single-dose oral toxicity test in Sprague Dawley rats was performed. Genotoxicity was assessed using a bacterial reverse mutation test with Salmonella typhimurium (strains TA98, TA100, TA1535, and TA1537) and Escherichia coli WP2 uvrA, an in vitro mammalian chromosome aberration test using Chinese hamster lung cells, and an in vivo mammalian erythrocyte micronucleus test using Hsd:ICR (CD-1) SPF mouse bone marrow. After LK1 administration (2,500 mg/kg), there were no LK1-related body weight changes or necropsy findings. The reverse mutation test showed no increased reverse mutation upon exposure to 5,000 ㎍/plate of the LK1 powder, the maximum tested amount. The chromosome aberration test and micronucleus assay demonstrated no chromosomal abnormalities and genotoxicity, respectively, in the presence of the LK1 powder. The absence of physiological findings and genetic abnormalities suggests that LK1 powder is appropriate as a candidate biomass to be used as a safe food ingredient.

Mutagenicity Study of Purified Bee Venom (Apis mellifera L.) by the Bacterial Reverse Mutation Assay (세균을 이용한 정제봉독의 복귀돌연변이시험)

  • Han, Sang Mi;Hong, In Phyo;Woo, Soon Ok;Kim, Se Gun;Jang, Hye Ri
    • Journal of Food Hygiene and Safety
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    • v.32 no.3
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    • pp.228-233
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    • 2017
  • The aim of the current study was to examine genotoxicological safety of purified bee venom (Apis mellifera L.) The bacterial reverse mutation in Salmonella typhimurium (TA100, TA1535, TA98, and TA1537) and Escherichia coli (WP2 uvrA) were evaluated with purified bee venom at concentrations of 0, 1.5, 5, 15, 50, 150, and $500{\mu}g/plate$. Purified bee venom was negative in Ames test with both in the presence and absence of rat liver microsomal enzyme. According to these results, we concluded that purified bee venom did not cause bacterial reverse mutation. The safety of the purified bee venom at practical doses needs to be further evaluated in in vivo genotoxicity assays.

Safety of Nano-sized Bee Pollen in both In-vitro and In-vivo Models (생체 외 및 생체 내 실험조건에서 나노화 벌 화분의 안전성 규명)

  • Pyeon, Hae-In;So, Soojeong;Bak, Jia;Lee, Seunghyun;Lee, Seungmin;Suh, Hwa-Jin;Lim, Je-Oh;Kim, Jung-Woo;Kim, Sun Youn;Lee, Se Ra;Lee, Yong Hyun;Chung, Il Kyung;Choi, Yun-Sik
    • Journal of Life Science
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    • v.28 no.5
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    • pp.605-614
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    • 2018
  • Bee pollen has an outer wall which is resistant to both acidic and basic solutions and even the digestive enzymes in the gastrointestinal tract. Therefore, the oral bioavailability of bee pollen is only 10-15%. A previous study reported on wet-grinding technology which increased the extraction of active ingredients from bee pollen by 11 times. This study was designed to investigate the safety of wet-ground bee pollen. First, a single dose of wet-ground bee pollen was tested in both rats and beagle dogs at dosages of 5, 10, and 20 g/kg and 1.5, 3, and 6 g/kg, respectively. In rats, compound-colored stools were found in those administered 10 g/kg or more of wet-ground bee pollen. In beagle dogs, 6 g/kg of wet-ground bee pollen induced diarrhea in one male for four hours. However, no obvious clinical signs were found through the end of the experiment in rats and beagle dogs. In addition, no histological abnormality was found in all animals. The data indicates that a single dose of up to 20 g/kg of wet-ground bee pollen is safe. Next, the genetic toxicity of nano-sized bee pollen was tested. This study employed a bacterial reverse mutation test, a micronucleus assay, and a chromosomal aberration assay. In the micronucleus assay, there was no genetic toxicity up to the dosage of 2 g/kg. There was also no genetic toxicity in the bacterial reverse mutation test and chromosomal aberration assay. This data provides important information in developing nano-sized bee pollen into more advanced functional foods and herbal medicines.