• 제목/요약/키워드: Activity assay

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In Vitro Antioxidant Activity Profiles of ${\beta}$-Glucans Isolated from Yeast Saccharomyces cerevisiae and Mutant Saccharomyces cerevisiae IS2

  • Song, Hee-Sun;Moon, Ki-Young
    • Food Science and Biotechnology
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    • 제15권3호
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    • pp.437-440
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    • 2006
  • To explore the possible usefulness of ${\beta}$-glucans as natural antioxidants, the antioxidant profiles of ${\beta}$-glucan, extracted from Saccharomyces cerevisiae KCTC 7911, and water soluble and insoluble mutant ${\beta}$-glucan, isolated from yeast mutant S. cerevisiae IS2, were examined by five different in vitro evaluation methods: lipid peroxidation value (POV), nitric oxide (NO), 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical, reducing power, and ${\beta}$-carotene diffusion assay. The antioxidant activities of all ${\beta}$-glucans evaluated in POV test were comparable to or better than that of the known antioxidant, vitamin C. Remarkably, the ${\beta}$-glucan and water insoluble mutant ${\beta}$-glucan possessed 2.5-fold more potent activity than vitamin C at a dosage of 2 mg. Although vitamin C showed 100-fold greater activity than all ${\beta}$-glucans in NO and DPPH tests for measuring the radical scavenging capacity, all ${\beta}$-glucans revealed higher radical scavenging activity than the known radical scavenger, N-acetyl-L-cysteine (NAC), in DPPH test. The water insoluble mutant ${\beta}$-glucan had 2.6- and 5-fold greater antioxidative activity than water soluble ${\beta}$-glucan in NO and DPPH tests, respectively, showing that all ${\beta}$-glucans were able to scavenge radicals such as NO or DPPH. While all ${\beta}$-glucans revealed lower antioxidant profiles than vitamin C in both reducing power activity and ${\beta}$-carotene agar diffusion assay, the ${\beta}$-glucan and water insoluble mutant ${\beta}$-glucan did show a marginal reducing power activity as well as a considerable ${\beta}$-carotene agar diffusion activity. These results confirmed the potential usefulness of these ${\beta}$-glucans as natural antioxidants.

식물 특정효소저해제의 생물활성 조사에 의한 신규제초제 작용점 탐색 (Searching of Possible Target Enzymes for Herbicide Development using Commercial Plant-Specific Inhibitors)

  • 황인택;최정섭;박상희;이관휘;이병회;홍경식;조광연
    • 농약과학회지
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    • 제5권1호
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    • pp.36-45
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    • 2001
  • 본 연구는 새로운 제초제 후보물질을 탐색하기 위하여 식물특이적 효소 저해제로 알려진 107개 기존 화합물에 대하여 생물활성을 조사하였다. Germination test, seedling assay, wheat leaf disc assay, cyanobacteria assay, whole plant assay를 통하여 15종의 저해제를 선발하였고 이들은 34종 효소를 저해하는 것으로 확인되었다. 이들 화합물 중에서 phenylhydrazine, purine, o-phenanthroline, oleylamine, 7,8-benzoquinoline, aminooxyacetic acid, dicyclohexylcarbodiimide 등은 성체를 이용한 온실 실험에서 높은 제초활성을 나타내었다. 7,8-benzoquinone, 8-hydroxyquinoline, 2,2'-dipyridyl 및 o-phenanthroline 등은 피, 벼, 토마토의 발아를 $1.25{\sim}5{\mu}M$의 농도에서도 억제하였다. 7,8-benzoquinoline, cyanuric fluoride, 4-methylpyrazole, tranylcypromine, oleylamine과 trifluoperazine 등은 $30{\sim}100{\mu}M$ 농도에서 cyanobacteria의 생육을 저해하였다. Dicyclohexyl carbodiimide와 chlorpromazine은 $100{\mu}M$ 농도에서 wheat leaf disc의 백화현상을 유기시켰다. 이상과 같이 생물학적 활성을 갖는 식물 특이적 효소저해제들은 신규제초제 후보물질을 선발하기 위한 새로운 대상효소로 이용될 수 있을 것으로 생각된다.

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Screening of Oriental Herbal Medicines for Antibacterial Activities

  • Bae, O-Sung;Hwang, Jae-Ock;Ahn, Duk-Kyun;Woo, Eun-Rhan;Seo, Seon-Hee;Kim, Hyoung-Ja;Park, Ho-Koon
    • Natural Product Sciences
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    • 제4권1호
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    • pp.32-37
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    • 1998
  • The water extracts of oriental herbal medicines which have been clinically used to treat bacterial infections in Korea were screened for in vitro antibacterial activity by the paper disc assay method. Two Gram positive bacteria, Staphylococcus aureus SG511, Bacillus subtilis ATCC 6633 and two Gram negative bacteria, Escherichia coli 055, Pseudomonas aeruginosa 9027 were used as test organisms. Among 83 of the extracts tested, 25 were active against Staphylococcus aureus SG511, 9 were active against Bacillus subtilis ATCC 6633, while none showed inhibitory activity against Eschelichia coli 055 and Pseudomonas aeruginosa 9027. Among them, Hwangyonhaedoktang plus hwangyon, Chongwisan, and Ssangbaksan showed remarkably potent antibacterial activity.

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7-Chloro-4-nitro-benzo[1,2,5]oxadliazole 1-oxide의 CDK4 활성저해 (Inhibition of CDK4 activity by 7-chloro-4-nitro-benzo[1,2,5]oxadiazole 1-oxide)

  • 전용진;고종희;연승우;김태용
    • 약학회지
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    • 제50권1호
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    • pp.52-57
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    • 2006
  • The activation of cyclin dependent kinase 4 (CDK4) is found in more than half of all human cancers. Therefore CDK4 is an attractive target for the development of a novel anticancer agent. For mass screening of CDK4 inhibitor, we set up in vitro kinase assay for CDK4 activity using a cyclin D1-CDK4 fusion protein, which is constitutively active and exhibits enhanced stability. From the screening of representative compound library of Korea Chemical Bank, we found that 7-chloro-4-nitro-benzo[1,2,5]oxadiazole 1-oxide (FBP-1248) selectively inhibited CDK4 activity in vitro by ATP competitive manner. This compound prevented the phosphorylation of retinoblatsoma tumor suppressor protein, Rb, and inhibited cell growth through cell cycle arrest. In summary, we developed an efficient assay system for CDK4 activity in vitro and identified the CDK4 inhibitory compound, FBP-1248.

Disease Suppressive Mechanisms of Antagonistic Bacteria against Phytophthorn capsici causing Phytophthora Blight of Pepper

  • Kim, Hye-Sook;Kim, Ki-Deok
    • 한국식물병리학회:학술대회논문집
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    • 한국식물병리학회 2003년도 정기총회 및 추계학술발표회
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    • pp.93.2-94
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    • 2003
  • In our previous studies, we selected three antagonistic bacteria, KJ1R5, KJ2C12, and KJ9C8 against Phytophthora capsici, the casual agent of Phytophthora blight of pepper. For elucidating production, root colonization, and total microbial activity were investigated. The dual culture assay was accomplished to elucidate existence of antibiotics. In this assay, any antagonistic bacteria did not inhibit growth of six important fungal plant pathogens, suggesting that these antagonists do not produce antibiotics. root surface or rhizosphere soil colonizations were examined with spontaneous rifampicin-resistant mutants equal to antagonistic ability of wild types. KJ2C12 colonized consistently rhizosphere soil while yellowish colonies of KJ1R5 and KJ9C8 well colonized root surfaces and rhizosphere soil. Total microbial activity in pots treated with the antagonistic bacteria was measured using fluorescein diacetate hydrolysis. total microbial activity of three antagonistic bacteria treatments was significantly higher than that of buffer-treated control until 4days after treatment. However, total microbial activity of treatment of three antagonistic bacteria decreased after 7 days. These results indicate that the antagonistic bacteria, KJ1R5 and KJ9C8 colonized and protected roots well against Phytophthora blight of pepper through competition of infection courts, especially competitions.

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Protective Effect of Crataegi Fructus Extract on the Neurotoxicity Induced by Reactive Oxygen Species in Cultured C6 Glioma Cell

  • ;유선미
    • 대한의생명과학회지
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    • 제14권1호
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    • pp.27-32
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    • 2008
  • To clerify the antioxidant effect of Crataegi Fructus (CF) extract on reactive oxygen species (ROS), The C6 glioma cells were treated with various concentrations of hydrogen peroxide ($H_2O_2$). The $H_2O_2$-induced neurotoxicity was measured by XTT assay for the cell viability. For the protective effect of CF extract on the cytotoxicity induced by $H_2O_2$, cell viability, lactate dehydroganase (LDH) activity, and the inhibitive activity of lipid peroxidation of CF extract were performed. In this study, $H_2O_2$ decreased cell viability dose- and time-dependent manners and increased LDH activity compared with the control. In the protective effect on $H_2O_2$, CF extract increased cell viability and decreased LDH activity on $H_2O_2$-induced cytotoxicity, lipid peroxidation by FTC assay. From these results, It is suggested that $H_2O_2$ was highly toxic on cultured C6 glioma cells, and also, CF extract showed the protective effect on $H_2O_2$-mediated cytotoxicity.

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DNA손상 및 돌연변이에 대한 명지버섯의 방어효능 (The Protective Effects of Ganoderma lucidum on the DNA Damage and Mutagenesis)

  • 이길수;공석경;최수영
    • Biomolecules & Therapeutics
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    • 제11권2호
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    • pp.139-144
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    • 2003
  • Ganoderma lucidum is commonly known as medically potent mushroom, which has been widely used in China and other oriental countries for the treatment of various diseases, including cancer. In this report, we investigated the anti-oxidant and protective effect of Ganodema lucidum extract (GLE) against the DNA damage induced by free radical and U.V. In the assay of cell growth inhibition, the inhibitory cell growth rate induced by hydroxyl radical was dose-dependently decreased by GLE. This results support that GLE has a detoxifying activity against cytotoxicity of hydroxyl radical in E. coli cell. GLE also protected ColE1 plasmid DNA damage in the concentration of 200$\mu\textrm{g}$ per reaction on the DNA fragmentation assay. The nuclear tailing by hydrogen peroxide in single cell gel electrophoresis(SCGE) was decreased by GLE in the concentration of 50$\mu\textrm{g}$/ml. These data indicate that Ganoderma lucidum has an anti-oxidative activity to hydrogen peroxide. The mutation rate after irradiation of U.V. was reduced by 50$\mu\textrm{g}$/ml GLE and total number of Rif (Rifampicin) resistant mutants was decreased in a concentration dependent manner when added the GLE exogenously in a culture media. According to the results, it is likely that GLE has not only an anti-oxidative activity to hydroxyl radical but also an anti-mutagenic activity to U.V. mutagenesis.

서양민들레와 흰민들레 추출물의 Nitric Oxide 생성억제 및 소거 활성과 Tyrosinase 저해 활성 (Nitric Oxide Production Inhibitory and Scavenging Activity and Tyrosinase Inhibitory Activity of Extracts from Taraxacum officinale and Taraxacum coreanum)

  • 임도연;이경인
    • 한국약용작물학회지
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    • 제19권5호
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    • pp.362-367
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    • 2011
  • The study was conducted to investigate functional materials as skin whitening and anti-inflammatory agent from Taraxacum officinale and Taraxacum coreanum. The total polyphenol and flavonoid content in the ethanol extract of Taraxacum officinale were found to be 64.07mg/g and 32.46mg/g, respectively. In tyrosinase inhibitory activity, the hot water extract of Taraxacum coreanum was higher than the other extracts. However, in nitric oxide (NO) scavenging ability, the ethanol extract of Taraxacum coreanum was higher than the other extracts. the ethanol extract of Taraxacum coreanum showed strong NO production inhibitory effect in lipopolysaccharide (LPS)-stimulated Raw 264.7 cell. In the cell viability measurement by MTT assay and the lactate dehydrogenase (LDH) assay against L929 cell, the extracts were exhibited fine cell viabilities and normal LDH release levels as nontoxic result in sample concentration of $250{\sim}1000{\mu}g/m{\ell}$. As a result, the ethanol extract and the hot water extract of Taraxacum coreanum could be applicable to functional materials for anti-inflammatory and skin whitening related fields, respectively.

유자의 항 Influenza 바이러스 A형 활성에 관한 연구 (Study on the Anti-influenza Virus A type Activity of Citrus junos)

  • 김호경;고병섭;전원경
    • 생약학회지
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    • 제31권1호
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    • pp.82-86
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    • 2000
  • To evaluate anti-influenza virus activity of 113 specimens of Korean traditional medicine both water and methanol extracts were examined using haemagglutination inhibition test. The water extract from Citrus junos was found to inhibit influenza virus A/Taiwan/l/86(H1N1). The survival rates of virus were determined by in situ cellular enzyme-linked immunosorbent assay. The water extract of Citrus junos was fractionated by chromatographic separating using Amberlite XAD-4, 40% MeOH and 60% MeOH layer had antiviral activity. The half inhibition concentration $(IC_{50})$ of 40% MeOH layer on survival of influenza virus was $MIC>361.5{\mu}g/ml$ and $IC_{50}$ value of fr. 40-4 fractionated from 40% MeOH layer was $677.19{\mu}g/ml$. These results suggested that the fractions of Citus junos have potent anti-influenza A virus activity.

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항고형암제의 활성평가를 위한 in vitro 삼차원 암세포 배양계의 확립 (Establishment of in vitro 3-Dimensional Tumor Model for Evaluation of Anticancer Activity Against Human Solid Tumors)

  • 이상학;이주호;구효정
    • Journal of Pharmaceutical Investigation
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    • 제34권5호
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    • pp.393-399
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    • 2004
  • For the efficient determination of activity against solid tumors, an in vitro tumor model that resembles the condition of in vivo solid tumors, is required. The purpose of this study was to establish a rapid culture method and viability assay for an in vitro 3-dimensional tumor model, multicellular spheroid (MCS). Among 12 human cancer cell lines, a few cell lines including DLD-1 (human colorectal carcinoma cells) formed fully compact MCS which was adequate for in vitro viability assay. DLD-1 MCS showed steady growth reaching $700\;{\mu}m$ diameter after 11 day culture. DLD-1 cells grown as MCS showed significant increase in $G_0/G_1$ phase compared to the monolayer cells (73.9% vs 45.7%), but necrotic regions or apoptotic cells were not observed. The cells cultured as MCS showed resistance to 5-FU (10.3 fold higher $IC_{50}$) compared to monolayers, however, tirapazamine (a hypotoxin) showed similar activity in both culture systems. In summary, MCS may be a valid in vitro model for activity screening of anticancer agents against human solid tumors and also exploitable for studying molecular markers of drug resistance in human solid tumors.