• Title/Summary/Keyword: Activity assay

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Fibrinolytic, thrombin inhibitory, anti-oxidative and anti-inflammatory activities of Pleurotus ferulea (아위버섯 추출물의 혈전용해, 트롬빈저해, 항산화 및 항염증 활성)

  • Kim, Eun-Jung;Kim, Jun-Ho
    • Journal of Mushroom
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    • v.13 no.1
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    • pp.30-36
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    • 2015
  • Our study investigated the fibrinolytic, thrombin inhibitory, anti-oxidative and anti-inflammatory activities of the water extract and solvent fractions isolated from Pleurotus ferulea. Fibrinolytic activity was investigated using the fibrin plate method. Thrombin inhibitory activity was used to analyze thrombin inhibitor assay. The DPPH assay was used to estimate anti-oxidative activity. Inhibition of NO production was measured for anti-inflammatory activity in LPS-activated murine RAW 264.7 macrophage cells. An MTS assay was used to evaluate the effects of the water extract and solvent fractions isolated from Pleurotus ferulea on cell viability. Our results showed the fibrinolytic activity to be strong in the ethyl acetate fraction at 1.33 plasmin units. The ethyl acetate fraction also showed high thrombin inhibitory activity at 94.45%. The anti-oxidative activity of the water extract was 37.01% and the anti-inflammatory activity of the chloroform fraction was 98.13%. These findings suggest that Pleurotus ferulea's extract and fractions could be applicable in the development of functional foods for the treatment and prevention of cardiovascular diseases.

Effects of DSG on Osteoblastic Cell from Rat Calvariae in the Presence of Dexamethasone (단치소요산가미방이 Dexamethasone 처리한 랫드의 두개골 세포에 미치는 영향)

  • Park, Jong-Hyeong;Hwang, Gwi-Seo
    • Journal of Society of Preventive Korean Medicine
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    • v.10 no.2
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    • pp.19-30
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    • 2006
  • It is well known that glucocorticoid may induce osteoporosis as its side effect in long-term therapy. The inhibition of osteoblast by glucocorticoid is also recognized as its action mechanism of decreased bone formation. In this study, the effect of DSG, Danchisoyosangamibang, on the differentiation and function of osteoblastic cells was investigated. The osteoblastic cells were isolated from rat calvariae using collagenase treatment. The cell counting, enzyme activity assay, MTT assay, collagen content assay were done to determine the cell proliferation, intracellular alkaline phosphatase (ALP) activity, bone martrix production, and cell apoptosis. DSG enhanced the cell proliferation after the culture for 10 days. ALP activity and total protein synthesis, and intracelluar collagen synthesis were increased time dependently when the cells were treated with DSG in the presence of dexamethasone. And, DSG restored calvarial cell function decreased by dexamethasone.

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Decrease of Activity of Estrogenic Chemicals by Chlorination (염소산화에 의한 에스트로겐성 화학물질의 활성저감)

  • Lee, Byoung-cheun;Lee, Sang-hyup;Kamei, Tasuku;Magara, Yasumoto
    • Journal of Korean Society of Water and Wastewater
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    • v.19 no.1
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    • pp.98-105
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    • 2005
  • The effects of chlorination on the elimination of three estrogenic chemicals such as $17{\beta}$-estradiol (E2), nonylphenol (NP) and bis-phenol A (BPA) were investigated using yeast two-hybrid assay (YTA), estrogen receptor competition assay (ER-CA), and high-performance liquid chromatography/mass spectrometer (LC/MS). Results of YTA, ECA and the analysis of LC/MS indicated that the estrogenic activity of above mentioned three endocrine disruptors were significantly reduced as the result of chlorination. The decrease in estrogenic activity paralleled with decrease in estrogenic chemicals under the influence of free chlorine. One common characteristic of estrogenic chemicals is the presence of a phenolic ring. Considering that a phenolic ring is likely to undergo some sort of transformation in aqueous chlorination solution, the above mentioned results may be applied to the rest of the other estrogenic chemicals in natural waters.

In vitro Effects of the Complex of Korean Citrus junos and Medicinal Herbs on the Cell Growth in CCD-986sk Human Fibroblast Cell Line

  • Park, Kap-Joo
    • Korean Journal of Environmental Biology
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    • v.22 no.1
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    • pp.83-88
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    • 2004
  • In order to investigate whether or not CCD-980sk cell line can be affected by Korean Citrus junos and medicinal herbs, we examined the MTT assay when we treated Korean Citrus junos and medicinal herbs in CCD-986sk human fibroblast cell line. The samples that added complex of grinded extracts of Korean Citrus junos and boiling-water extracts from Korean medicinal herbs were tested toy cell proliferation activity by means of a modification of the MTT assay. Among mixture of Citron 3 (Citron 3, less mellowed citron which was ripened for three months) and boiling-water extracts, the group Citron 3+Phellinus linteus showed significantly strong cell proliferation activity. And among mixture of Citron 4 (Citron 4, completely mellowed citron which was ripened for four months) and boiling-water extracts, the group Citron 4+Cordyceps militaris and Citron 4+Phellinus linteus showed significantly strong cell proliferation activity, respectively. These results suggest that complex of Korean Citrus junos and medicinal herbs could be an excellent candidate toy protection of human skin aging.

A Colorimetric Microplate Assay Method for High Throughput Analysis of Lipase Activity

  • Choi, Suk-Jung;Hwang, Jung-Min;Kim, Sung-Il
    • BMB Reports
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    • v.36 no.4
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    • pp.417-420
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    • 2003
  • The present work describes a colorimetric microplate assay for lipase activity based on the reaction between 5,5'-dithiobis(2-nitro benzoic acid) (DTNB) and the hydrolysis product of 2,3-dimercapto-1-propanol tributyrate (DMPTB). Reaction mixtures containing DTNB, DMPTB, and lipase were prepared in microplate wells, and the absorbance at 405nm was recorded after incubation at $37^{\circ}C$ for 30 min. A linear relationship was obtained in the range of 0.1-1 U of lipase activity by this method. The reaction conditions were also optimized for the range of 0.01-0.1 U or 1-10 U. When assaying crude tissue extracts, the reaction of DTNB with non-specific reducing agents created a major source of error. However, this error was corrected by the use of blank samples that did not contain DMPTB.

A Generic Time-resolved Fluorescence Assay for Serine/threonine Kinase Activity: Application to Cdc7/Dbf4

  • Xu, Kui;Stern, Alvin S.;Levin, Wayne;Chua, Anne;Vassilev, Lyubomir T.
    • BMB Reports
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    • v.36 no.4
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    • pp.421-425
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    • 2003
  • The serine/threonine protein kinase family is a large and diverse group of enzymes that are involved in the regulation of multiple cellular pathways. Elevated kinase activity has been implicated in many diseases and frequently targeted for the development of pharmacological inhibitors. Therefore, non-radioactive antibody-based kinase assays that allow high throughput screening of compound libraries have been developed. However, they require a generation of antibodies against the phosphorylated form of a specific substrate. We report here a time-resolved fluorescence assay platform that utilizes a commercially-available generic anti-phosphothreonine antibody and permits assaying kinases that are able to phosporylate threonin residues on protein substrates. Using this approach, we developed an assay for Cdc7/Dbf4 kinase activity, determined the $K_m$ for ATP, and identified rottlerin as a non-ATP competitive inhibitor of this enzyme.

Screening of Hyaluronidase Inhibitory Activity Using a Microplate Assay (Microplate방법을 이용한 Hyaluronidase 저해 활성 검색)

  • Jeong, Sei-Joon;Kim, Na-Young;Ahn, Nyeon-Hyoung;Kim, Youn-Chul
    • Korean Journal of Pharmacognosy
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    • v.28 no.3
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    • pp.131-137
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    • 1997
  • The aqueous and methanolic extracts of 110 crude drugs were screened for hyaluronidase inhibitory activity using a microplate assay. Among them, MeOH extract of 15 crude drugs inhibited more than 80% of hyauluronidase activity at the concentration of 5mg/ml. The active principles of Anemarrhenae Rhizoma, Rhei Rhizoma, Ephedrae Herba, Pteropi Faeces and Ginseng Radix alba were transferred into organic solvents.

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In vivo Antitumor Activity of Mansonone E isolated from Ulmus davidiana var. japonica NAKAI (느릅나무의 근피로부터 분리한 Mansonone E의 항암효과)

  • Cho, Choa-Hyung;Yoon, Won-Ho;Lee, Keyong-Ho
    • Korean Journal of Pharmacognosy
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    • v.35 no.3 s.138
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    • pp.199-202
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    • 2004
  • A highly potent antitumor sesquiterpene O-naphthoquionone has been isolated from Ulmus davidiana, which has been traditionally used as the folk medicine. The structure of this compound was established on the basis of spectral data obtained from UV, IR, MS, and NMR spectrometry, and determined as mansonone E. This compound showed potent antitumor activity on in vivo assay, hollow fiber and xenograft assay.

Effects of JY on Osteoblastic Cell from Rat Calvariae in the Presence of Glucocorticoid (자혈양근탕(滋血養筋湯)이 부신피질호르몬에 의해 억제된 조골세포 기능에 미치는 영향)

  • Choi, Jeong-Sin;Hwang, Gwi-Seo
    • Journal of Society of Preventive Korean Medicine
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    • v.12 no.2
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    • pp.197-206
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    • 2008
  • The inhibition of osteoblast by glucocorticoid is recognized as its action mechanism of decreased bone formation. In this study, the effect of JY, Jahyulyangkeuntang, on the differentiation and mineralization of osteoblastic cells was investigated in the presence of dexamethasone. The cell counting, enzyme activity assay, MTT assay, collagen content assay were done to determine the cell proliferation, alkaline phosphatase(ALP) activity, bone martrix production, and cell apoptosis. JY enhanced the cell proliferation after the culture for 10 days. ALP activity and total protein synthesis, and intracellular collagen synthesis were increased when the cells were treated with JY. And JY restored calvarial cell function decreased by dexamethasone.

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A Modification of A Microbiological Assay of Vitamin $B_{12}$ In Fermented Vegetables

  • Lim, Sook-Ja
    • Journal of Nutrition and Health
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    • v.15 no.3
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    • pp.202-211
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    • 1982
  • A modification of a microbiological assay of vitamin $B_{12}$ was made and used to determine the vita­min levels during kimchi fermentation. A cyanide-buffer solution of pH 6.0 replaced the metabisufite-buffer specified in the A.O.A.C. method. The vitamin $B_{12}$ activity was decreased by blending kimchi samples for 5 minutes and retained the activity by steaming for 10 minutes before blending. The alkali hydrolysis of kimchi at pH 12.0 for 30 minutes at $121^{\circ}C$ was sufficient to destroy the vitamin $B_{12}$ and permit the detection of analogs with the same assay organism. Vitamin $B_{12}$ reached a maximum of 47ng/100g during the fermentation of kimchi ${15}\;4^{\circ}C.$ Inoculation of the kimchi with Propionibacterium shermanii (ATCC 13673) increased the vitamin production to a maximum of 102ng/100g at 1 week of fermentation. Soy flour (0.5%) or beef extract (0.05%), which were regarded as protein sources, added to the inoculated kimchi further increased the vitamin $B_{12}$ activity to 197 and 203ng/100g.

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