• Title/Summary/Keyword: bioassay system

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A Comparative Study of the Induction by Positive Control of Revertant Colonies in Salmonella typhimurium TA98 and SCE in Human Lymphocytes (Ames test와 자매염색분체교환분석법(SCE)에서의 positive control에 의한 유전독성 비교연구)

  • 임흥빈;이영구;이동욱;김용태
    • Journal of the Korean Society of Tobacco Science
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    • v.13 no.1
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    • pp.19-26
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    • 1991
  • Ames test using special strains which are histidine requiring mutant of Salmonella typhimufium , is widely utilized as short-term bioassay to evaluate the genotoxicity of chemicals. This method requires the liver microsome(5-9 fraction) to provide mammalian metabolism of the compounds, Therefore, the mutagenic potency of the chemicals is affected by not only the intrinsic properties of them but also the efficiency of the in vitro microsomal activation system. For this reason, the complex mixtures such as environmental pollutants from occupational sources, natural products or cigarette smoke condensates(CSC) , might be often appeared the false results. Induction of sister chromatid exchange(SCE) in cultured cells is known as another sensitive and powerful tool for the measurement of genotoxicity and the method has also an advantage which is able to apply to the in vitro and in vivo systems. In the present study, the inducibilities of revertant colonies in tester strain TA98 and SCE in human Iymphocytes by positive controls and total particulated materials(TPM) obtained from various brand(domastic and imported) cigarettes were compared in order to investigate whether the results in Ames test are in agreement with those in SCE analysis. The mutagenic activities of well known mutagens such as B(a)P showed excellent dose-response in the both methods although the induction mechanism was different each other, but cyclophosphamide resulted such effect only in SCE analysis. Most TPM tested showed a similar pattern in the mutagenic activities in those methods. However, only two(one imported brand and one domestic sample cigarettes) among the TMP obtained from various cigarettes appeared the higher induction in SCE than Ames test.

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Inhibitory Activity of Nitric Oxide Synthase and Peroxynitrite Scavenging Activity of Extracts of Perilla frutescens (들깨 잎 추출물의 Nitric Oxide Synthase 저해활성 및 Peroxynitrite 소거활성)

  • Kim, Jae-Yeon;Kim, Ji-Sun;Jung, Chan-Sik;Jin, Chang-Bae;Ryu, Jae-Ha
    • Korean Journal of Pharmacognosy
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    • v.38 no.2 s.149
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    • pp.170-175
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    • 2007
  • Activated microglia by neuronal injury or inflammatory stimulation overproduce nitric oxide (NO) by inducible nitric oxide synthase (iNOS) and reactive oxygen species (ROS) such as superoxide anion, resulting in neurodegenerative diseases. The toxic peroxynitrite (ONOO$^-$), the reaction product of NO and superoxide anion further contributes to oxidative neurotoxicity. We tried to evaluate the effects of two kinds of varieties of Perilla frutescens var japnica Hara on the NO production in lipopolysaccharide (LPS)-activated microglia. The perilla cultivars of Namcheondeulkkae (NC) and Boradeulkkae (BR) were developed by pure line from the local variety and by a cross between 'deulkkae' and 'chajogi', respectively. Spirit, hexane, chloroform and butanol fractions of the leaves of NC and BR inhibited the production of NO in LPS-activated microglia. The fractions of BR showed stronger activity than NC and the spirit extracts was the most potent in both cultivars. The solvent fractions of BR suppressed the expression of protein and mRNA of iNOS in LPS-activated microglial cells. Moreover, the extracts of NC and BR showed the activity of peroxynitrite scavenging in cell free bioassay system. These results imply that Namcheondeulkkae and Boradeulkkae might have neuroprotective activity through the inhibition of NO production by activated microglial cells and peroxynitrite scavenging activity.

Construction of a Baculovirus Hyphantria cunea NPV Insecticide Containing the Insecticidal Protein Gene of Bacillus thuringiensis subsp. kurstaki HD1

  • Lee, Hyung-Hoan;Moon, Eui-Sik;Lee, Sung-Tae;Hwang, Sung-Hei;Cha, Soung-Chul;Yoo, Kwan-Hee
    • Journal of Microbiology and Biotechnology
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    • v.8 no.6
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    • pp.685-691
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    • 1998
  • Baculovirus Hyphantrin. cunea nuclear polyhedrosis virus (HcNPV) insecticide containing the insecticidal protein (ICP) gene from Bacillus thuringiensis subsp. kurstaki HD1 was constructed using a lacZ-HcNPV system. The ICP ($\delta$-endotoxin) gene was placed under the control of the polyhedrin gene promoter of the HcNPV. A polyhedrin-negative virus was derived and named ICP-HcNPV insecticide. Then, the insertion of the ICP gene in the ICP-HcNPV genome was confirmed by Southern hybridization analysis. Polyacrylamide gel electrophoresis (PAGE) analysis of the Spodoptera frugiperda cell extracts infected with the ICP-HcNPV showed that the ICP was expressed in the insect cells as 130 kDa at 5 days post-infection. The ICP produced in the cells was present in aggregates. When extracts from the cells infected with the ICP-HcNPV were fed to 20 Bombyx mori larvae, the following mortality rate was seen; 8 larvae at 1 h, 10 larvae at 3 h, and 20 larvae at 12 h. These data indicate that the B. thuringiensis ICP gene was expressed by the baculovirus insecticide in insect cells and there was a high insecticidal activity. The biological activities of the recombinant virus ICP-HcNPV were assessed in conventional bioassay tests by feeding virus particles and ICP to the insect larvae. The initial baculovirus insecticide ICP-HcNPV was developed in our laboratory and the significance of the genetically engineered virus insecticides is discussed.

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Subchronic Toxicity of Herbicide Butachlor in Fish , the Medaka (Oryzias latipes) (제초제 Butachlor의 송사리에 대한 아급성(亞急性) 독성(毒性))

  • Shin, Chun-Chul;Lee, Sung-Kyu;Roh, Jung-Ku
    • Korean Journal of Environmental Agriculture
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    • v.4 no.2
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    • pp.118-125
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    • 1985
  • To establish an evaluation system of aquatic toxicity of chemicals at no-effect level, flow through and early life stage toxicity test were performed on a freshwater fish, the medaka (Oryzias latipes). The characteristics of medaka as a bioassay organism for the chronic toxicity test were discussed. Maximum acceptable toxicant concentration(MATC) of butachlor for the madaka in soft water was estimated using survival, growth, and reproduction as indicators of toxic effects. During a 3-month period, the fry of medaka were exposed to butachor concentrations ranging from 0.16 to 0.0l mg/liter and the DO concentration, temperature, and pH in the exposure chamber were measured to check the test condition. The highest concentration showed slight decrease of growth rate in medaka and reduced hatchability of spawning egg. Survival, growth, and reproductive success of adults in butachlor concentration of 0.04 and 0.01 mg/liter were not different from those of the control. The MATC was estimated to be between 0.04 and 0.16 mg/liter for medaka.

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숙주범위가 넓어진 재조합 Baculoviruses의 특성 및 병원성 검정

  • Kim, Hye-Sung;Woo, Soo-Dong;Kim, Woo-Jin;Choi, Jae-Young;Jin, Byung-Rae;Oh, Hyun-Woo;Lee, Youn-Hyung;Kang, Seok-Kwon
    • Microbiology and Biotechnology Letters
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    • v.25 no.4
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    • pp.359-366
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    • 1997
  • The usefulness of host range expanded recombinant viruses for economical viral insecticide and expression vector system has been studied. Host range expanded recombinant viruses, RecS-B6 and RecB-8, constructed by cotransfection of Bombyx mori nuclear polyhedrosis virus (BmNPV) and Autographa californica NPV (AcNPV), and a host range expanded AcNPV recombinant, Ac-BH, constructed by substitution of the 0.6Kb fragment of the BmNPV helicase gene were compared. The restriction enzyme digestion patterns showed that RecS-B6 and RecB-8 had expanded host ranges by genomic recombination and were more similar to genome of AcNPV than that of BmNPV. SDS-PAGE and PCR analysis showed that the polyhedrin gene of RecS-B6 and RecB-8 was derived from BmNPV genomic DNA. The morphology of polyhedra of recombinant viruses showed a slight difference between the two host cells, Sf and BmN cells, indicating that the morphology of polyhedra was influenced by host cells. The bioassay data for insect larvae showed that Ac-BH, compared to wild type viruses, had superior pathogenicity against Bombyx mori larvae but inferior pathogenicity against Spodoptera exigua larvae. Although the pathogenicity was lower than that of wild type viruses in both larvae, RecS-B6 showed the pathogenicity in both larvae. These results suggested that Ac-BH was a less useful economical insecticide than random genomic recombinant virus RecS-B6.

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Inhibitory Effects of the Stem Bark Extract of Eucommia ulmoides on the Proliferation of Human Tumor Cell Lines (두충(杜沖) 추출물의 암세포증식 저해효과)

  • Choi, Yeon-Hee;Seo, Jee-Hee;Kim, Jung-Sook;Heor, Jung-Hee;Kim, Seong-Kie;Choi, Sang-Un;Kim, Young-Sup;Kim, Young-Kyoon;Ryu, Shi-Yong
    • Korean Journal of Pharmacognosy
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    • v.34 no.4 s.135
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    • pp.308-313
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    • 2003
  • A bioassay-guided fractionation of the stem bark extract of Eucommia ulmoides Oliver (Eucommiaceae) led to the isolation of three iridoid constituents, genipin (1), geniposide (3), geniposidic acid (4) as well as $({\pm})-guaiacylglycerol$ (2) and fatty acid mixtures as active ingredients of the extract responsible for the antitumoral property. The EtOAc soluble part and BuOH soluble part of the extract demonstrated a potent inhibition on the proliferation of cultured human tumor cells such as A549 (non small cell lung), SK-OV-3 (ovary), SK-MEL-2 (melanoma), XF498 (central nerve system) and HCT-15 (colon) in vitro, whereas the remaining water soluble part exhibited a poor inhibition. The intensive investigation of the EtOAc soluble part and BuOH soluble part of the extract yielded genipin, guaiacylglycerol, geniposide, geniposidic acid and large amounts of fatty acid mixtures as active components.

Issues in 2-year Long-term in vivo Carcinogenicity Assay (2년 장기 발암성 검색법의 이슈)

  • Son, Woo-Chan;Kim, Bae-Hwan;Jang, Dong-Deuk;Han, Beom-Seok;Kim, Jong-Choon;Lee, Je-Bong;Shin, Jin-Sup;Kim, Hyoung-Chin
    • The Korean Journal of Pesticide Science
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    • v.8 no.4
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    • pp.239-254
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    • 2004
  • It has been debating about conducting and interpretating of 2-year rodent carcinogenicity bioassay. Although some criticisms arising in usefulness, it has been still known that long-term carcinogenicity studies using rodents would be the only assay system to predict any possible human risks, which would not be replaced. Both regulatory agencies and academies have developed some assay models, however, there have been controversy whether those study designs and interpretations are based on sound scientific rationale and validated data. Such kinds of issues including choice of species/strain, dose level selection, duration of study, number of animals per group, historical control data, monitoring parameters, terminal investigations, peer review, statistics, alternative assay models, interpretation of neoplastic lesions, and risk assessments, were reviewed.

In vitro Screening of Medicinal Plants with Estrogen Receptor Modulation Activity (생약의 여성호르몬 수용체 조절 활성 검색)

  • Lee, Chang-Min;Kang, Se-Chan;Oh, Joa-Sub;Choi, Han;Li, Xue-Mei;Lee, Jae-Hyun;Lee, Mi-Hyun;Choung, Eui-Su;Kawk, Joung-Hwan;Zee, Ok-Pyo
    • Korean Journal of Pharmacognosy
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    • v.37 no.1 s.144
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    • pp.21-27
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    • 2006
  • Yeast based estrogenicity assay is the simplest and useful for the assay and the discovery of novel estrogenic substances in natural specimens, The estrogen receptor(ER) modulation activity of 50% EtOH extracts of 101 traditional medicinal herbs was assessed using a recombinant yeast assay system with both a human estrogen receptor expression plasmid and a receptor plasmid. Among them, 14 species proved to be active. Pureariae Flos (flower of Puerraria thunbergiana BENTH.) had the highest estrogenic relative potency$(7.75{\times}10^{-3})$ $(EC_{50}=9.39\;{\mu}g/ml)$. The $EC_{50}$ value of $17{\beta}-estradiol$ used as the positive control was $0.073\;{\mu}g/ml)$ (Relative Potency=1.00). There results demonstrated that some of the traditional medical herb may be useful in the therapy of estrogen replacement.

Isolation and Partial Characterization of Phytotoxic Mycotoxins Produced by Sclerotinia sp., a Potential Bioherbicide for the Control of White Clover(Trifoliorum repens)

  • Hong, Yeon-Kyu;Lee, Bong-Choon;Jung, Won-Kwon;Bae, Soon-Do;Park, Sung-Tae;Uhm, Jae-Youl
    • The Plant Pathology Journal
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    • v.20 no.1
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    • pp.52-57
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    • 2004
  • Sclerotinia sp. (isolate BWC98-105) causes stem blight and root rot in Leghum sp., and is presently being evaluated as a potential mycoherbicide for the control of Trifoliorium repens. Bioassays have shown that Sclerotinia sp. produces phytotoxic substance which is biologically active against T. repens. Two biologically active compounds, designated as compoundsI and II, were produced in vitro from the culture filtrate of BWC98-105 isolate Sclerotium sp. Compounds I and II were purified by means of liquid-liquid extraction and $C_{18}$ open column chromatography (300 ${\times}$ 30 mm, i.d). To determine the purity, the purified compounds were analyzed by RP-HPLC. The analytical RP-HPLC column was a TOSOH ODS-120T (150 ${\times}$ 4.6 mm i.d, Japan), of which the flow rate was set at 0.7 mL/min using the linear gradient solvent system initiated with 15 % methanol to 85 % methanol for 50 min with monitoring at 254 nm. Under these RP-HPLC conditions, compounds I and II eluted at 3.49 and 4.13 min, respectively. Compound II was found to be most potent and host specific. However, compound I had a unique antibiotic activity against phytopathogenic bacteria like bacterial leaf blight (Xanthomonas oryzae) on rice, where it played a less important role in producing toxicity on T. repens. No toxin activity was detected in the water fraction after partitioning with several organic solvents. However, toxin activity was detected in the ethyl acetate and butanol fractions. In the leaf bioassay using compound II, the disease first appeared within 4-5 h as water soaked rot, which subsequently developed into well-defined blight affecting the whole plant.

Screening of Endogenous Maize (Zea mays) Substances Enhancing Auxin-induced Inward Curvature in Coleoptilar Slits (안쪽으로 굽어지는 자엽초 박편의 옥신 반응을 촉진하는 옥수수(Zea mays) 내생물질의 탐색)

  • Park, Woong-June
    • Journal of Life Science
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    • v.16 no.5
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    • pp.859-865
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    • 2006
  • When thin slits (e.g., $1mm{\times}10mm$) of maize (Zea mays) coleoptiles were floated on a buffer, they spontaneously curved outward because of unbalanced tissue tension between inner and outer faces. Exogenously applied auxin induced inward curvature of the thin strip of the maize coleoptile in a dose-dependent manner. This bioassay system was used to screen endogenous substances that work together with auxin. In methanol extract of maize coleoptiles including the leaves inside, Active fractions that promote the auxin-induced inward curvature of maize coleoptile slices were found. The curvature-enhancing activity of the extract was not related to energy supply. The active substances were adsorbed to $C_{18}$ cartridges even at pH 10 and eluted in two fractions by 50% and 80% methanol. These substances were named as Curvature-Enhancing Factor-1 (CEF-1) and Curvature-Enhancing Factor-2 (CEF-2), respectively. The CEF-2 was resolved on a reversed phase $C_{18}$ column by HPLC.