• Title/Summary/Keyword: Micronucleus.

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Anti-mutagenic and Anti-septic Effects of $\beta$-glucan from Aureobasidium pullulans SM-2001 (흑효모유래 $\beta$-glucan의 패혈증 치료효과 및 항돌연변이 활성 평가)

  • Ku, Sae-Kwang
    • Korean Journal of Oriental Medicine
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    • v.15 no.3
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    • pp.75-82
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    • 2009
  • Anti-mutagenic and anti-septic effects of $\beta$-1,3/1,6-glucan from Aureobasidium pullulans SM-2001 were evaluated on the on the cyclophosphamide (CPA)-cecal ligation puncture (CLP) and CPA-treated mice. To induce immunosuppression and mutagenicity, 150 and 110 mg/kg of CPA were single intraperitoneally injected at 3 or 1 day before CLP or initial $\beta$-glucan administration. In CLP animals, the cecum was mobilized and ligated below the ileocecal valve, punctured through both surfaces twice with a 22-gauge needle. 125 mg/kg of $\beta$-glucan were dissolved in saline and subcutaneously or orally administered in a volume of 10 ml/kg (of body weight), 4 times, 12 hrs intervals from 6 hrs after CLP or 1 day after second dose of CPA. After treatment of $\beta$-glucan, the mortalities were observed in CPA-CLP model, and the appearance of a micronucleus is used as an index for genotoxic potential in CPA model. As results of CPA-CLP sepsis, all animals (9/9, 100%) in CPA-CLP control were dead within 2 days after CLP. In addition, increase of the number of bone marrow MNPCEs indicated mutagenicity were also observed by treatment of CPA. However, $\beta$-glucan treatment effectively inhibited the mortalities in CPA-CLP, and it also reduced the CPA treatment-related mutagenicity, respectively. These results indicated that $\beta$-glucan has effective anti-septic and anti-mutagenic effects and can be used as an agents for treating sepsis and mutagenicity related to high-dose chemotherapy or radiotherapy. However, further studies should be conducted to observe more detail action mechanisms of it's anti-septic and anti-mutagenic effects.

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Genetic Toxicity Test of Glycidol by Ames, Micronucleus, Comet Assays and Microarray Analysis

  • Kim, Ji-H.;Kim, Ki-Y.;Kwon, Kyoung-J.;Go, Seo-Y.;Min, Kyung-N.;Lee, Woo-S.;Park, Sue-N.;Sheen, Yhun-Y.
    • Biomolecules & Therapeutics
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    • v.14 no.4
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    • pp.240-245
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    • 2006
  • The primary use for glycidol is as a stabilizer in the manufacture of vinylpolymers, however, it is also used as an intermediate in the production of pharmaceuticals, as an additives for oil and synthetic hydraulic fluids, and as a diluting agent is same epoxy resins. In this study, we have carried out in vitro genetic toxicity test of glycidol and microarray analysis of differentially expressed genes in response to glycidol. The result of Ames test showed mutations with glycidol treatment in base substitution strain TA1535 both with and without exogenous metabolic activation. Likewise, glycidol showed mutations in frame shift TA98 both with and without exogenous metabolic activation. The result of COMET assay in L5178Y cells with glycidol treatment showed DNA damage both with and without exogenous metabolic activation. Glycidol increased micronuclei in CHO cells both with and without exogenous metabolic activation. 150 Genes were selected as differentially expressed genes in response to glycidol by microarray analysis and these genes would be candidate biomarkers of genetic toxic action of glycidol.

In Vitro Assessment of Cytotoxicity and Mutagenicity of Rock Wool Fibers (암면에 의한 세포독성 및 변이원성의 실험실적 평가)

  • Hong, Yun-Chul;Lee, Kwan-Hee
    • Journal of Preventive Medicine and Public Health
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    • v.30 no.3 s.58
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    • pp.555-566
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    • 1997
  • This study was carried out to evaluate the cytotoxicity of rock wool fibers(RWFs) such as cell division disturbance, chromosomal and DNA damage, and mutagenicity using cultured cells. RWFs were the man made mineral fibers. In order to find the correlation between the cytotoxicity of RWFs and the phagocytic capacity of cells, the phagocytic processes were observed using scanning electron microscope. Cell division disturbance by RWFs was evaluated by the formation of multinucleated giant cells. The chromosomal damage was evaluated by the micronucleus formation. For the evaluation of oxidative DNA damage, 8-hydroxy-2'-deoxyguanosine (8-OH-dG) formation was measured utilizing calf thymus DNA. Mutagenicity was determined by the point mutation of HGPRT and the effect of RWFs on cell transformation was also observed. 1. Compared with the results of chrysotile, RWFs were no or little effect on the cell growth according to the results done by the tests of cell proliferation inhibition and relative plating efficiency. 2. The frequency of multinucleated giant cell formation was increased by the treatment of RWFs and it was dose-dependent. However, the effect of RWFs was weaker than that of chrysotile. 3. The number of micronuclei formed in the RWFs treated cells was between those of cells treated with chrysotile and those of untreated cells. 4. The 2 fold increase in the formation of 8-OH-dG in calf thymus DNA was observed in the cells treated with RWFs in the presence of $H_2O_2$. On the other hand, chrysotile had no effect on the 8-OH-dG formation. 5. RWFs had no effect on the HGPRT point mutation and cell transformation. These results showed that RWFs could induce chromosomal damage, cell division disturbance and oxidative DNA damage in the RWFs treated cells.

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Evaluation of Genotoxicity and 14-day Repeat Dose Toxicity of Water Extract of the Mixture of Natural Plants, Anemarrhena and Phellodendron (천연식물자원 지모와 황백피 혼합 수추출물의 안전성평가)

  • Chung, Young-Shin;Park, Cheol-Beom
    • Journal of Food Hygiene and Safety
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    • v.29 no.4
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    • pp.383-390
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    • 2014
  • The safety of a new natural plant composition (ADP) was assessed on the genotoxicity study and 14-day repeat dose toxicity study. ADP contains a mixed water extract obtained from the mixture of Phellodendron cortex (Phellodendron amurense) and Anemarrhena rhizoma (Anemarrhena asphodeloides), and poses the contractile properties mediated by alpha-adrenoceptor of the prostate and urethra as well as antioxidant and anti-inflammatory properties. In order to evaluate genetic safety, in vivo micronucleus test was performed in ICR mice orally administered with three dose levels of 1250, 2500, 5000 mg/kg body weight, and vehicle and positive control. In the 14 days study, Sprague-Dawley rats were treated with ADP at the dose levels of 500, 1000, 2000 mg/kg once a day, and clinical signs, body weights, hematology, serum biochemistry, necropsy findings and organ weights were monitored and examined. In experimental results, ADP treatment, compared with vehicle control, did not induce the micronucleated erythrocytes from mouse bone marrow. In the 14 days study, any significant and toxicological differences in all measurements of parameters were not observed in ADP treatment groups of animals, compared with vehicle treatment. The No-Observed-Adverse-Effect-Level (NOAEL) of ADP in the 14 days study was determined to be greater than 2000 mg/kg/day in both sexes.

Antigenotoxic Effects of Synurus deltoides Extract on Benzo[a]pyrene Induced Mutagenesis

  • Ham, Seung-Shi;Han, Hong-Sik;Park, Kun-Pyo;Oh, Deog-Hwan
    • Preventive Nutrition and Food Science
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    • v.2 no.2
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    • pp.162-166
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    • 1997
  • This study investigated the antigenotoxic effects of Synurus deltoides extract on the mutagenesis induced by benzo[a]pyene(B[a]P). About 80% and 90% antimutagenic effects were observed in the presence of over 200$\mu\textrm{g}$/plate of methanol extract of Synurus deltoides against Salmonella typhimurium TA98 and TA100 induced by B[a]P, respectively. The methanol extract itself did not induce an increased frequency of micronucleated polychromatic erythrocytes(MNPCE) irrespective of the sampling time(up to 72h), while the treatment with benzo[a]pyrene(B[a]P) at 150mg/kg significantly increased (p<0.05) the incidence of MNPCE. The strongest relative frequency of MNPCE was observed at 36h after injection of B[a]P and the most significant reduction (p<0.05) in the frequencies of MNPCE was occurred at the feeding of the methanol extract to mice 12 h before injection of B[a]P. The most significant reduction (p<0.05) with 48% was observed in the frequencies of MNPCE when 50 mg/kg of the methanol extract was given to the mice 12h before injection of B[a]P, while the strongest relative frequency inhibition was 54% at the multiple feeding of 5mg/kg of the methanol extract e time every day for 5 days on th frequencies of MNPCE induced by 150 mg/kg of B[a]P. These results indicate that the methanol extract of Synurus deltoides have a strong modulatory effect on benzo[a]pyrene induced MNPCE.

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Screening and prevention of the mutagenicity for fishes accordind to cookery and storage (어류의 가열조리 및 보존에 의해 생성되는 변이원성 물질의 정량적 해석과 제어법)

  • 홍이진;이준경;구성자
    • Korean journal of food and cookery science
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    • v.16 no.6
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    • pp.652-662
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    • 2000
  • This study was performed to measure the mutagenicity of fish by cooking and storage. Mutagenicity of the fish extract was measured by Ames test(Salmonella typhimurium reversion assay with TA 100) in vitro and by micro-nucleus test in vivo. The fish samples screened in this study were white fish(Trichiurus, Croaker, Salted Croaker) and red fish(Saury pike, Mackerel, Yellowtail, Salmon). The number of revertants of red fish were significantly higher than that of white fish. And the mutagenicity of mackerel was higher than other red fish, so followed experiment was made by using the extract of mackerel. Mutagenicity of the samples cooked on microwave oven was the lowest, whereas there was no significant difference between the samples cooked on gas grill and the ones on electric grill. In the presence of S9 mixture, the methanol extract of mackerel showed 2∼4 times high values of mutagenicity in comparison with the extract without S9. The extract of mackerel cooked with various vegetable juices showed inhibitory effects on the mutagenicity in the order of green tea, ginger, and radish. Also, the number of revertants was increased in the stored samples. Mutagenicity of the samples stored in the refrigerator was higher than that of the freezer. In micronucleus test, the methanol extract treated with vegetable juice inhibited micro-nucleus formation in bone marrow by cyclophosphamide in the order of ginger, green tea, and radish. In TBA test, there was a tendency that TBA values were increased as the storage time increased. Also, the rancidity of sample were stored in the refrigerator was higher value than sample stored in the freezer. Samples cooked on microwave oven showed the highest value in rancidity. When the antioxidant effect of vegetable juice was measured by electron donating ability(EDA) of mackerel cooked with vegetable juice to DPPH, the samples treated with onion showed the highest value of EDA(%), and the samples treated with green tea, ginger and cabbage also showed the antioxidant effect.

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Studies on the Genetic Toxicity of Guh Sung Y.L.S.-95 (목초액 (거성 Y.L.S-95)의 유전독성에 관한 연구)

  • Lee Soo-Yong;Li Guang-Yong;Yin Hu-Quan;Jung Eun-Jung;Kim Youn-Su;Lee Hye-Young;Lee Byung-Hoon
    • Journal of Food Hygiene and Safety
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    • v.21 no.2
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    • pp.107-112
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    • 2006
  • Guh Sung Y.L.S-95 (GS95) is a kind of polyacidic solution, which contains acetic acid as a main component. We investigated in the present study tile genetic toxicity of GS95 according to the standard operation procedure from Korean Institute of Toxicology. In the Salmonella typhimurium reverse mutation assay using TA1535, TA1537, TA98 and TA100, GS95 did not induce mutation up to $5,000{\mu}g/plate$. GS95 did not induce chromosome aberration in Chinese hamster lung fibroblast in the concentration range between 1.25 and 5 mg/mL. In the rodent micronucleus assay, the frequency of micronucleated polychromatic erythrocyte in GS95 treated mice were not increased up to 5,000 mg/kg compared to the vehicle treated mice. Taken all these data together, GS95 was proven to be nongenotoxic in the concentration ranges tested.

Nuclear Anomalies, Chromosomal Aberrations and Proliferation Rates in Cultured Lymphocytes of Head and Neck Cancer Patients

  • George, Alex;Dey, Rupraj;Bhuria, Vikas;Banerjee, Shouvik;Ethirajan, Sivakumar;Siluvaimuthu, Ashok;Saraswathy, Radha
    • Asian Pacific Journal of Cancer Prevention
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    • v.15 no.3
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    • pp.1119-1123
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    • 2014
  • Head and neck cancers (HNC) are extremely complex disease types and it is likely that chromosomal instability is involved in the genetic mechanisms of its genesis. However, there is little information regarding the background levels of chromosome instability in these patients. In this pilot study, we examined spontaneous chromosome instability in short-term lymphocyte cultures (72 hours) from 72 study subjects - 36 newly diagnosed HNC squamous cell carcinoma patients and 36 healthy ethnic controls. We estimated chromosome instability (CIN) using chromosomal aberration (CA) analysis and nuclear level anomalies using the Cytokinesis Block Micronucleus Cytome Assay (CBMN Cyt Assay). The proliferation rates in cultures of peripheral blood lymphocytes (PBL) were assessed by calculating the Cytokinesis Block Proliferation Index (CBPI). Our results showed a significantly higher mean level of spontaneous chromosome type aberrations (CSAs), chromatid type aberration (CTAs) dicentric chromosomes (DIC) and chromosome aneuploidy (CANE UP) in patients (CSAs, $0.0294{\pm}0.0038$; CTAs, $0.0925{\pm}0.0060$; DICs, $0.0213{\pm}0.0003$; and CANE UPs, $0.0308{\pm}0.0035$) compared to controls (CSAs, $0.0005{\pm}0.0003$; CTAs, $0.0058{\pm}0.0015$; DICs, $0.0005{\pm}0.0003$; and CANEUPs, $0.0052{\pm}0.0013$) where p<0.001l. Similarly, spontaneous nuclear anomalies showed significantly higher mean level of micronuclei (MNi), nucleoplasmic bridges (NPBs) and nuclear buds (NBUDs) among cases (MNi, $0.01867{\pm}0.00108$; NPBs, $0.0156{\pm}0.00234$; NBUDs, $0.00658{\pm}0.00068$) compared with controls (MNi, $0.00027{\pm}0.00009$; NPBs, $0.00002{\pm}0.00002$; NBUDs, $0.00011{\pm}0.00007$).The evaluation of CBPI supported genomic instability in the peripheral blood lymphocytes showing a significantly lower proliferation rate in HNC patients ($1.525{\pm}0.005552$) compared to healthy subjects ($1.686{\pm}0.009520$) (p<0.0001). In conclusion, our preliminary results showed that visible spontaneous genomic instability and low rate proliferation in the cultured peripheral lymphocytes of solid tumors could be biomarkers to predict malignancy in early stages.

Redescription of Three Newly Recorded Gastrostyla Ciliates (Ciliophora: Spirotrichea: Stichotrichida) with Morphological Variations from Korea (한국미기록 Gastrostyla 섬모충 (Ciliophora: Spirotrichea: Stichotrichida) 3종의 형태적 변이와 재기재)

  • Jo Jong O;Kim YeonUk;Shin Mann Kyoon
    • Animal Systematics, Evolution and Diversity
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    • v.21 no.1
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    • pp.45-56
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    • 2005
  • Three species of Gastrostyla ciliates collected from the sewage treatment plant and a puddle in the valley from Korea were identified as Gastrostyla minima Hemberger, 1985, G. steinii Engelmann, 1862 and G. setifera (Engelmann, 1862). The description was based on the observation of living specimens, protargol impregnated specimens and biometric analysis. The morphological variations among the populations of these species were investigated with morphometry. These species have not been reported in Korea and their diagnostic characteristics are as follows: G. minima normally has two oval macronuclei (Ma) with one spherical micronucleus (Mi) respectively, continuous ventral cirral row (VCR) with additional one postoral ventral cirrus (poVC), five transverse cirri (TC), six dorsal kineties (DK) with broken 4th kinety, and cortical granules. G. steinii has four oval Ma with three to five Mi, discontinuous VCR with additional one poVC, four TCs, six DKs, and no cortical granules. G. setifera has two oval Ma with one spherical Mi respectively, five TCs which distinctly separate in two groups, six DKs, discontinuous VCR with additional two poVCs, and no cortical granules.

Interaction between odontoblast and bio-calcium phosphate cement reinforced with chitosan (치아모세포와 키토산으로 강화된 생체 적합 칼슘인산시멘트와의 상호작용)

  • Chun, Byung-Do;Kim, Sung-Won;Lee, Sung-Tak;Kim, Tae-Hoon;Lee, Jung-Han;Kim, Gyoo-Cheon;Kim, Yong-Deok;Kim, Uk-Kyu
    • Journal of the Korean Association of Oral and Maxillofacial Surgeons
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    • v.37 no.5
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    • pp.415-420
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    • 2011
  • Purpose: Calcium phosphate cement (CPC) is one of many useful materials for restoring tooth defects, periodontium and maxillofacial area. Chitosan is a biodegradable material that has been shown to promote the growth and differentiation of osteoblasts in culture. This study examined the interaction between odontoblasts and bio-calcium phosphate cement reinforced with chitosan. Materials and Methods: $5{\times}10^3$ odontoblastic cells were seeded into each well. Various concentrations of bio-calcium phosphate cement reinforced with chitosan (10, 20, 50, 100, 200, 500 ${\mu}g$/ml, 1, 2, 4 mg/ml) were diluted and added to the wells. The well was incubated for 24 h, 48 h and 72 h. After incubation, the number of cells was assessed to determine the cell viability. A cytokinesis-block micronucleus assay and chromosomal aberration test were carried out to estimate the extent of chromosomal abnormalities. Microscopic photographs and RT-PCR were performed to examine the adhesion potential of bio-calcium phosphate cement reinforced with chitosan. Results: Bio-CPC-reinforced chitosan did not show significant cytotoxicity. The number of damaged chromosomes in the cells treated with Bio-CPC-reinforced chitosan was similar to that in the control cells. There was no significant increase in the number of chromosomal aberrations in the Bio-CPC reinforced chitosan exposed cells. Microscopic photographs and RT-PCR confirmed the adhesive potential of bio-CPC reinforced chitosan to odontoblasts. Conclusion: Bio-CPC-reinforced chitosan did not affect the odontoblastic cell viability, and had no significant cytotoxic effect. Bio-CPC-reinforced chitosan showed adhesive potential to odontoblasts. These results are expected form the basis of future studies on the effectiveness of dental restorative materials in Bio-CPC reinforced with chitosan.