• Title/Summary/Keyword: Functional assay

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Studies on the development of enzyme linked immuno-sorbent assay (ELISA) for hepatitis B surface antigen (HBsAg) by monoclonal antibodies of different affinity constants

  • Kim, Gye-Won;Hong, Sung-Youl;Shin, Soon-Cheon;Lee, Sung-Hee;Kim, Won-Bae
    • Archives of Pharmacal Research
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    • v.10 no.1
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    • pp.18-24
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    • 1987
  • Mouse monocolonal antibodies to Hepatitis B surface antien (HBsAg) were prepared and their functional capabilities tested by the method of solid phase enzyme linked immuno sorbent assay (ELISA). HBsAg binding studies inicated that one monoclonal antibody 6E-1-1 bound more HBsAg at a faster rate than the other monoclonal antibodies. Also, for the binding inhibition studies with the selected monoclonal antibody 6E-1-1, one monoclonal antibody 8D-3-6 didn't exhibit binding inhibition for HBsAg. Then, a simultaneous ELISA method was developed for the immunodiagnosis of HBsAg. Different combinations of two monoclonal antibodies as solid phase and horseradish peroxidase (HRPO) labeled phase were studied. The combination of monoclonal antibody of higher affinity constant (6E-1-1) immobilized in a solid phase and monoclonal antibody of lower affinity constant (8D-3-6) as a HRPO laeled phase was more sensitive when two monoclonal antibodies of different affinity constants for HBsAg were prepared.

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Immunomodulatory Effects of Eckol, a Pure Compound of Ecklonia Cava, on Dendritic Cells

  • Kim, Mi-Hyoung;Joo, Hong-Gu
    • IMMUNE NETWORK
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    • v.6 no.4
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    • pp.199-203
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    • 2006
  • Background: Eckol purified from Ecklonia cava, a brown alga has been known to have cytoprotective effects on some cell lines against oxidants and ionizing radiation. However, there is no study about the effects of eckol on immune cells. Methods: Bone marrow (BM)-derived dendritic cells (DCs) were used to demonstrate the immunomodulatory effects of eckol on DCs, such as viability, the expression of surface markers, allogeneic stimulating capacity using MTI, flow cytometric, $^3H$-thymidine incorporation assay. Results: Eckol did protect DCs against cytokine withdrawal-induced apoptosis in a concentration dependent manner based on MTT assay. And also, it increased the expression of MHC class II and CD86 (B7.2) molecules, maturation markers, on the surface of viable DCs gated in FACS analysis. Furthermore, eckol-treated DCs stimulated the proliferation of allogeneic $CD4^+$ T lymphocytes compared to imDCs in $^3H$-thymidine incorporation assay. $CD4^+$ T lymphocytes activated with eckol-treated DCs produced the larger amount of IFN-${\gamma}$ and IL-4 than those cells with imDCs. Conclusion: Taken together, we demonstrate in this study that eckol, a pure compound of Ecklonia cava, may modulate the immune responses through the phenotypic and functional changes of DCs.

Phenolic Compounds from Antioxidant Plant Materials and their Protective Effect on PC12 cells (항산화 식물의 페놀성화합물에 의한 PC12 세포보호 효과연구)

  • You, SoHyeon;Kim, Gun-Hee
    • Journal of the Korean Society of Food Culture
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    • v.33 no.1
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    • pp.86-94
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    • 2018
  • This study was designed to evaluate the antioxidant activities and protective effects on PC12 cells of the extract of Epimedium koreanum and its main constituents icariin and icariside I. After screening the seven identified flavonoid glycosides from E. koreanum through DPPH (2,2-diphenyl-1-picryl-hydrazyl-hydrate) assay, E. koreanum, Icariin and Icariside I exhibited significant effect on radical scavenging activity. E. koreanum, icariin and icariside I were examined using DPPH, ABTS (2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid)) and FRAP (ferric reducing ability power) assay. In all antioxidant assays, E. koreanum, icariin and icariside I showed high radical scavenging activities in a dose-dependent manner. Protective effects against $H_2O_2-induced$ PC12 cells were assessed with MTT assay. The results indicated that cell viability and protection on PC12 cells of icariside I and icariin increased dose dependently. These study results suggest that E. koreanum, icariin and icariside showed high antioxidant capacities and cell protective effects. Icariside I, one of the metabolites of icariin, may be a new and effective flavonoid compound as a functional component.

Haemocytes responses of the pearl oyster, Pincdata fucata, at different temperatures

  • Choi, Young-Joon;Hwang, Jee-Youn;Baeck, Gun-Wook;Kim, Mu-Chan;Park, Hyung-Jun;Choi, Byoung-Dae;Kang, Suk-Joong;Park, Chan-Il
    • Journal of fish pathology
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    • v.21 no.3
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    • pp.241-246
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    • 2008
  • The effects of temperatures on pearl oyster, Pincdata fucata were studied by evaluating some functional immune responses of the haemocytes. Water temperature is one of the most important factors in bivalve immune defense. Haemocytes comprise a primary line by inflammation, encapsulation and phagocytosis. These phagocytic abilities of haemocytes were observed in different temperatures. The number of the circulating haemocytes by migratory assay, phagocytic activities by MTT assay and reactive oxygen species production of haemocytes by CL assay were measured at different temperatures. Results showed that pearl oyster maintained at 20℃ and 25℃ displayed significantly higher values for all the measured immune parameters in comparison to maintained at 10, 15, and 30℃.

Antioxidant Activity of Vegetables or Fruits Extract in Mice (야채 및 과일추출물의 항산화작용)

  • Heo Chan;Kim Nam Yee;Kim Hyun Pya;Heo Moon Young
    • YAKHAK HOEJI
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    • v.49 no.3
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    • pp.249-254
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    • 2005
  • The ethanol extracts of the mixed vegetables (Bioactive Vegetables, BV) and the mixed fruits (Bioactive Fruits, BF) were evaluated for their in vivo antioxidant activities. Four weeks treatment of oral administration was performed to mice. A $KBrO_3$ as a potent oxidant was used to induce the oxidative stress for in vivo experiment. BV and BF were shown to possess the significant inhibitory effect of lipid peroxidation as measured by the level of malondialdehyde (MDA) formation although the potencies were not higher than those of well-known antioxidants such as vitamin C, trolox and quercetin. Furthermore, BV and BF inhibited DNA damage assessed by single cell gel electrophoresis (comet assay) and reduced the micronucleated reticulocyte (MNRET) formation of peripheral blood. Antioxidants tested also revealed potent inhibitory activities higher than BV and BF. These antigenotoxic activity profiles were similar to that of abovementioned inhibition of lipid peroxidation. Therefore, BV and BF having mild antioxidant activity as functional food candidates may be useful natural antioxidants by the inhibiting of lipid peroxidation and the protecting oxidative DNA and chromosomal damage.

Physiological Activity of Extracts of Bark from Ulmus davidiana var. japonica and its fractions (유근피 추출물 및 분획의 생리활성)

  • Yang, Sun a;Kim, A young;Pyo, Byoung sik;Kim, Sun min
    • Korean Journal of Pharmacognosy
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    • v.50 no.2
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    • pp.112-117
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    • 2019
  • In this study, extracts of bark from Ulmus davidiana var. japonica and its fractions were investigated the antioxidative, antibacterial and tyrosinase inhibition activity for physiological activity towards functional applications. In the measurement of DPPH radical scavenging activity, the ethyl acetate fraction showed higher radical scavenging ability than others. Moreover, in the tyrosinase inhibition assay, showed that the ethyl acetate fraction has good inhibition effects. Results of the DPPH radical scavenging and tyrosinase inhibition activity are related with the total polyphenol concentrations of ethyl acetate fraction. In antibacterial activity used to find out by utilizing the disc diffusion assay, chloroform fraction showed strong effect against Staphylococcus aureus and S. epidermidis. These results are related with the flavonoid contents of chloroform fraction. On the other hand, in the L929 cell viability measurement by MTT assay, the hexane, butanol and aqueous fraction treated at high concentration were showed cytotoxicity. But the others samples were exhibited a moderate viabilities. As a result of investigated the antioxidant and tyrosinase inhibition activity, the ethyl acetate fraction could be applicable for cosmetics related fields. And the chloroform fraction showed significant antibacterial activity against S. aureus and S. epidermidis.

A Novel Rapid Fungal Promoter Analysis System Using the Phosphopantetheinyl Transferase Gene, npgA, in Aspergillus nidulans

  • Song, Ha-Yeon;Choi, Dahye;Han, Dong-Min;Kim, Dae-Hyuk;Kim, Jung-Mi
    • Mycobiology
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    • v.46 no.4
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    • pp.429-439
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    • 2018
  • To develop a convenient promoter analysis system for fungi, a null-pigment mutant (NPG) of Aspergillus nidulans was used with the 4'-phosphopantetheinyl transferase (PPTase) gene, npgA, which restores the normal pigmentation in A. nidulans, as a new reporter gene. The functional organization of serially deleted promoter regions of the A. nidulans trpC gene and the Cryphonectria parasitica crp gene in filamentous fungi was representatively investigated to establish a novel fungal promoter assay system that depends on color complementation of the NPG mutant with the PPTase npgA gene. Several promoter regions of the trpC and crp genes were fused to the npgA gene containing the 1,034-bp open reading frame and the 966-bp 3' downstream region from the TAA, and the constructed fusions were introduced into the NPG mutant in A. nidulans to evaluate color recovery due to the transcriptional activity of the sequence elements. Serial deletion of the trpC and crp promoter regions in this PPTase reporter assay system reaffirmed results in previous reports by using the fungal transformation step without a laborious verification process. This approach suggests a more rapid and convenient system than conventional analyses for fungal gene expression studies.

Evaluation of Antioxidative activity of Korean Yam (Dioscorea batatas DECNE.) by n-Butanol and Ethyl Acetate Extracts

  • Duan, Yishan;Kim, Han-Soo;Kim, Gyeong-Hwuii
    • Journal of the Korean Applied Science and Technology
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    • v.32 no.2
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    • pp.312-319
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    • 2015
  • In this study, n-butanol and ethyl acetate extracts were prepared from raw yam (Dioscorea batatas DECNE.). Their antioxidative potencies were investigated employing various in vitro methods, such as ferrous ion chelating, ${\beta}$-carotene bleaching assay, lipid peroxidation inhibition and nitric oxide (NO) radical scavenging and nitrite scavenging activity. The n-butanol fraction was assayed to possess stronger antioxidant activity by ${\beta}$-carotene bleaching assay, lipid peroxidation inhibition and NO radical scavenging activity. However, ethyl acetate extract was more effective in chelating ferrous ion and scavenging nitrite. Based on the results obtained, yam is a potential active ingredient that could be applied in antioxidation as well as bio-health functional food to take a good part in prevention of human diseases and aging.

Immunostimulating Activity of Phellinus linteus Extracts to B-lymphcyte

  • Oh, Goo-Taeg;Han, Snag-Bae;Kim, Hwan-Mook;Han, Man-Woo;Yoo, Ick-Dong
    • Archives of Pharmacal Research
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    • v.15 no.4
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    • pp.379-381
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    • 1992
  • Phellinus linteus was examined on its immunostimulating activities using an in vitro imunization and plaque forming cell assay. When lymphocytes were exposed to the extract of Phellinus linteus, the number of antibody forming cell was increased. In in vitro plaque forming cell assay, the immunostimulating effect was about 4.8 and 5.0 times of unimmunized control in polyconal and T-independent antibody response, respectively. Especially, Phellinus linteus significantly increased the antigenicity of TNP-LPS used as T-independent antigen. But Phellinus linteus did now show a mitogenic effect on B-lymphcytes. These results suggest that immunostimulating activity of Phillinus lintues might be associated with a functional stimulation of B-lympohocyte involved in humoral immune response.

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A Quinolone Alkaloid, from the Aleurone Layer of Oryza sativa cv. Mihyangbyo, Inhibits Growth of Cultured Human Leukemia Cell

  • Chung, Ha-Sook
    • Preventive Nutrition and Food Science
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    • v.7 no.2
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    • pp.119-122
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    • 2002
  • Oryza sativa cv. Mihyangbyo is one of several recently developed varieties of rice; characterized by high levels of aromatic components, which may increase its sensory and nutritional properties. In conjunction with our continuing investigation of bioactive components of improved grain varieties, a quinolone alkaloid was isolated from the n-butanol soluble fraction of the aleurone layer of Oryza sativa cv. Mihyangbyo (Gramineae) through activity-guided fractionation and isolation. The compound exhibited moderate antineoplastic activity in a human leukemia cell line (U937) with an $IC_{50}$/ value of 118.1 ug/mL, based on the MTT(3-[4, 5]dimethylthiazol-2-yl]-2, 5-diphenyltetrazolium bromide) cell proliferation assay. The chemical structure of the functional compound was determined, based on physical and spectroscopic characteristics.