• Title/Summary/Keyword: enzyme linked immunosorbant assay

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Inhibition Enzyme-Linked Immunosorbent Assay (ELSIA) for Rapid Dection of Pseudomonas fluorescens in Raw Milk using IgY (난황 laY 항체의 Inhibition Enzyme-Linked Immunosorbant Assay(ELISA)법을 이용한 원유내 Pseudomonas fluorscens의 신속 검출 방법 개발)

  • 이승배;최석호;백두연
    • Food Science of Animal Resources
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    • v.20 no.3
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    • pp.231-235
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    • 2000
  • 원유에 존재하는 Ps. fluorescens의 균수를 신속하게 측정하기 위한 Inhibition Enzyme Linked Immunosorbant Assay (ELSIA)를 개발하기 위해 Ps, fluorescens(KCTC 2344)을 산란계에 접종하여 Ps.fluorescens에 대한 anti-Ps. fluorescens IgY 항체를 생산하고 그 항체의 역기를 ELISA로 측정한 결과 32일까지 항체 역가가 증가하였으며, 분리된 IgY 항체의 titer는 1:128,000으로 나타났다. Anti-Ps.fluorescens IgY 항체에대한 교차반응을 조사한 결과 그람양성균 Lactococcus faecalis, Staphylococcus aureus 뿐만 아니라 그람음성 균인 Achrombacter sal-monisida, Escherichia coil와도 교차반응을 거의 하지 않는 것으로 나타났다. Anti-Ps. fluo-rescens IgY 항체를 가지고 Inhibition ELISA 방법으로 Ps. fluorescens 균수를 신속한 측정할 수 있는 표준곡선을 작성한 결과 Ps. fluorescens균을 5.0$\times$$10^4$cfu/ml부터 5.0$\times$$10^{8}$cfu/ml 측정할 수 있는 것으로 나타났다.

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Production of Nitric Oxide and Erythropoietin in Serum of Newborn (신생아의 혈청내 Nitric Oxide와 Erythropoietin의 생성)

  • 정현기;김광혁
    • Journal of Life Science
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    • v.9 no.2
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    • pp.201-206
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    • 1999
  • Nitric oxide(NO) is a potent and selective pulmonary artery vasodilator. Erythropoietin(EPO) is produced in the kidney in response to reduced oxygen availability. In this study, blood samples were collected for determination of NO and EPO concentrations in 18 normal newborns and 16 newborns with respiratory distress syndrome(RDS). Serum was measured by Ding's method for NO concentration and by enzyme-linked immunosorbant assay for EPO concentration. Nitric ion concentration in serum was 14.9$\pm$3.2 $\mu$M in normal control group and 12.8$\pm$3.3 $\mu$M in RDS group. EPO concentration in serum was 16.2$\pm$3.4 mU/ml in normal control group and 21.2$\pm$5.4 mU/ml in RDS group. These results show the decrease of NO and increase of EPO in RDS newborn patients in comparison with normal newborns. Such imbalances may contribute to the development of several clinical symptoms.

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Seroprevalence of porcine reproductive and respiratory syndrom(PRRS) in Dangjin (당진지역 돼지생식기호흡기증후군(PRRS) 항체가 조사)

  • Kong, Shin-Koog;Lee, Gun-Taek;Lee, Kwan-Bok;Hong, Jun-Pyo;Kang, Soo-Jeong;Moon, Sun-Hwa
    • Korean Journal of Veterinary Service
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    • v.26 no.3
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    • pp.227-231
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    • 2003
  • The purpose of this study was sero-epidemiological survey of porcine reproductive and respiratory syndrom(PRRS) in Dangjin area. 411 samples from 26 pig farms were analyzed by enzyme linked immunosorbant assay (ELISA). The data indicate that 66% of the pigs and 92% of the farms showed sero-positives to the PRRS viruc. Sows showed 58% of sero-positive rate and fattening pigs showed 85% of seropositive rate. The rate of sero-positive in boars was 63%. No significant regional differences were detected in sero-epidemiological survey.

Effect of modified-Okbyungpoongsan on mast cell-mediated allergic responses in RBL-2H3 mast cells (가미옥병풍산(加味玉屛風散)의 비만세포에서의 알레르기 반응에 대한 효과 연구)

  • Jung, Jin-Ki;Park, Yong-Ki
    • The Korea Journal of Herbology
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    • v.26 no.4
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    • pp.1-7
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    • 2011
  • Objectives : In this study, we investigated the effect of modified-Okbyungpoongsan (mOP) on mast cell-mediated allergic response in basophilic leukemia cell line, RBL-2H3 mast cells. Methods : Cells were stimulated with anti-DNP-IgE after the treatment of DNP-HSA (AI/D), and then incubated with different concentrations of mOP (0.1, 0.2, 0.5 and 1 mg/$m{\ell}$) in RBL-2H3 cells. Cell toxicity was determined by WST-1 assay. The degranulation of mast cells was observed by microscope with toluidine blue staining and also the levels of beta-hexosaminidase, histamine and TNF-alpha were measured in culture supernatants by enzyme-linked immunosorbant assay. Results : mOP inhibited anti-DNP-IgE-imduced degranulation of mast cells in RBL-2H3 cells. mOP also significantly decreased the levels of histamine and inflammatory cytokine, TNF-alpha in RBL-2H3 cells, but slightly decreased the level of beta-hexosaminidase. Conclusions : These results indicate that mOP, an oriental prescription could be inhibit the allergic response through suppressing the mast cell activation.

Analysis of the Potent Platelet Glycoprotein IIb-IIIa Antagonist from Natural Sources

  • Kang, In-Cheol;Kim, Doo-Sik
    • BMB Reports
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    • v.31 no.5
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    • pp.515-518
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    • 1998
  • Adhesive interaction of the platelet glycoprotien IIb-IIIa (GP IIb-IIIa) with a plasma protein, such as fibrinogen, plays an important role in thrombosis and hemostasis. The specific sequence Arg-Gly-Asp (RGD) is critical for the binding of fibrinogen to platelet. To examine and characterize the GP IIb-IIIa antagonist from natural sources, we have developed a simple enzyme-linked immunosorbant assay (ELISA) system. The GP IIb-IIIa complex was purified to homogeneity from platelet Iysates by the combination of two affinity chromatographic methods using the synthetic RGD peptide (GRGDSPK)-immobilized Sepharose and wheat germ lectin-Sepharose. The synthetic peptide GRGDSP inhibits GP IIb-IIIa binding to immobilized fibrinogen with an $IC_{50}$ of $1.5\;{\mu}M$. Venoms of three different snake species and a Korean scolopendra extract have strong antagonistic activities for the binding of human fibrinogen to the platelet GP IIb-IIIa complex. The $IC_{50}$ values of the snake venom s and scolopendra were in the range of $5.5\;{\mu}g$ to $60\;{\mu}g$. These results provide meaningful information for developing antiplatelet agents.

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Production and Secretion of Human Interleukin-18 in Transgenic Tobacco Cell Suspension Culture

  • Sharma, Niti;Kim, Tae-Geum;Yang, Moon-Sik
    • Biotechnology and Bioprocess Engineering:BBE
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    • v.11 no.2
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    • pp.154-159
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    • 2006
  • Interleukin-18 (IL-18), otherwise known as interferon-gamma-inducing factor (IGIF), is one of several well characterized and important cytokines that contribute to host defenses. The complementary DNA (cDNA) of mature human interleukin-18 gene (hIL-18) was fused with the signal peptide of the rice amylase 1A gene (Ramy1A) and introduced into the plant expression vector under the control of a duplicated CaMV 35S promoter. The recombinant plasmid was transformed into tobacco (Nicotiana tabacum L. cv Havana) using the Agrobacterium-mediated transformation method. The integration of the hlL-18 gene into the genome of transgenic tobacco plants was confirmed by polymerase chain reaction (PCR) amplification and its expression was observed in the suspension cells that were derived from the transgenic plant callus by using Northern blot analysis. The hlL-18 protein was detected in the extracts of the transgenic callus and in the medium of the transgenic tobacco suspension culture by using immunoblot analysis. Based upon enzyme-linked immunosorbant assay (ELISA) results, the expression level of the hlL-18 protein approximated $166{\mu}g/L$ in the suspension culture medium. Bioassay results from the induction of $interferon-{\gamma}$ from a KG-1 cell line indicated that the hlL-18 secreted into the suspension culture medium was bioactive.

Development of transgenic rice lines expressing the human lactoferrin gene

  • Lee, Jin-Hyoung;Kim, Il-Gi;Kim, Hyo-Sung;Shin, Kong-Sik;Suh, Seok-Cheol;Kweon, Soon-Jong;Rhim, Seong-Lyul
    • Journal of Plant Biotechnology
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    • v.37 no.4
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    • pp.556-561
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    • 2010
  • Lactoferrin is an 80-kDa iron-binding glycoprotein that is found in high concentrations in human milk. Human lactoferrin (hLF) has several beneficial biological activities including immune system modulation and antimicrobial activity. In the present study, we devolved a method of hLF expression through introducing the hLF gene construct into Oriza sativa cv. Nakdong using the Agrobacterium-mediated transformation system. The expression of the hLF gene under the control of the rice glutelin promoter was detected in the seeds of transgenic rice plants. Transformed rice plants were selected on media containing herbicide(DL-phosphinothricin) and the integration of hLF cDNA was confirmed by Southern blot analysis. The expression of the full length hLF protein from the grains of transgenic rice plants was verified by Western blot analysis. The lactoferrin expression levels in the transformed rice grains determined by enzyme-linked immunosorbant assay accounted for approximately 1.5% of total soluble protein. Taken together, these data indicate that rice grains expressing hLF can be directly incorporated into infant formula and baby food.

Chloroform Fraction of Zingiberis Rhizoma Recens Modulates the Production of Inflammatory Mediators in LPS-stimulated BV2 Microglial Cells (생강 클로로포름 분획의 활성화된 뇌신경교세포(腦神經膠細胞)에서 염증반응 억제효과)

  • Seo, Un-Kyo;Jung, Hyo-Won;Park, Yong-Ki
    • The Korea Journal of Herbology
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    • v.23 no.3
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    • pp.73-83
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    • 2008
  • Objectives : The root of Zingiber officinale ROSC. (Zingiberis Rhizoma Recens; Ginger) has been widely used as one of folk remedies and food materials in many traditional preparations. Ginger is known as an effective appetite enhancer and anti-inflammatory agent. This study was performed to investigate the effect of ginger chloroform fraction (GCF) in microglia which play a central role on brain inflammation in neurodegenerative diseases. Methods : Dried ginger was extracted with 80% methanol, and then fractionated with chloroform. BV2 mouse microglial cells were cultured with different concentrations of GCF and then stimulated with LPS (1 ${\mu}g/m{\ell}$) at indicated times. The cell toxicity of GCF was determined by MTT assay. The concentrations of NO, PGE2 and cytokines were measured by Griess assay and enzyme-linked immunosorbant assay. The mRNA and protein expressions of iNOS, COX-2 and cytokines were determined by RT-PCR and Western blotting. The phosphorylation of three MAPKs (p38 MAPK, ERK1/2 and JNK) and $NF-{\kappa}B$ activation were determined by Western blotting. Results : GCF significantly inhibited LPS-induced production of inflammatory mediators, NO, $PGE_2$ and proinflammatory cytokines ($TNF-{\alpha}$ and $IL-1{\beta}$) in a dose-dependent manner. GCF attenuated LPS-induced expression of mRNA and protein of inflammatory enzymes, iNOS, COX-2 and proinflammatory cytokines through suppressing the phosphorylation of ERK1/2 and p38 MAPK and the activation of p65 $NF-{\kappa}B$ in BV2 cells. Conclusions : This study suggests that GCF may have an anti-inflammatory property through suppressing the inflammatory mediator production released by activated microglia after the brain injury.

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Inhibition of gene expression and production of iNOS and $TNF-{\alpha}$ in LPS-stimulated microglia by On-Bi-Tang (오령산가온비탕의 BV2에서 LPS로 유도된 nitric oxide 및 $TNF-{\alpha}$ 생성에 미치는 영향)

  • Son, Hye-Young;Kim, Sang-Woon;Jung, Sun-Ju;Jung, Hyo-Won;Yoon, Cheol-Ho;Park, Yong-Ki
    • The Korea Journal of Herbology
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    • v.21 no.2
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    • pp.135-142
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    • 2006
  • Objectives : On-Bi-Tang(OB) has been prescribed Chinese traditional medicine for the treatment of inflammatory disease such as chronic renal failure. In this study, we investigated the anti-inflammatory effect of OB extract in the BV2 murine microglial cells. Methods : After the water extract of OB was treated in BV2 cells, murine microglial line, the cell viability was measured by MTT assay. The production of nitric oxide (NO) and $TNF-{\alpha}$ was determined based on Griess reagent and enzyme linked immunosorbant assay (ELISA). mRNA expression of inducible nitric oxide synthase (iNOS) and $TNF-{\alpha}$ was analyzed by RT-PCR. Results : OB extract significantly inhibited the LPS-induced production of NO and TNF-a in BV2 cells. OB extract also suppressed the mRNA expression of iNOS and $TNF-{\alpha}$ in BV2 cells activated with LPS. Conclusion : These data suggests that OB extract may have the anti-inflammatory effect through the modulation of NO production and inflammatory cytokine such as $TNF-{\alpha}$.

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