• Title/Summary/Keyword: Protein-A gold

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Gold Sodium Thiomalate Inhibits iNOS Gene Expression in RAW 264.7 Macrophage: Differential Regulation by Gold Sodium Thiomalate and Sodium Salicylate (Gold Sodium Thiomalate에 의한 유도성 Nitric Oxide Synthase Gene의 발현억제: Gold 제제와 Sodium Salicylate의 차이점)

  • 임종호;배진영
    • Biomolecules & Therapeutics
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    • v.10 no.4
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    • pp.230-235
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    • 2002
  • Gold sodium thiomalate (GST, gold compound) is a widely used anti-arthritic, anti-rheumatic and anti-inflammatory drug that is considered a good alternative to sodium salicylate (NaSA) for individuals who cannot tolerate salicylates. Nitric oxide (NO) synthesized by inducible nitric oxide synthase (iNOS) has been implicated as a mediator of inflammation. Recent evidence suggests that anti-inflammatory effect of NaSA lies in the inhibition of iNOS, but nothing has been reported about the direct effect of iNOS expression by GST. The present study was designed to elucidate sequentially the action mechanisms of GST and NaSA on lipopolysaccharide (LPS) plus interferon-gamma (IFN-$\gamma$) induced iNOS expression in RAW 264.7 macrophages. Both GST and NaSA inhibited NO production and iNOS protein expression in a dose dependent manner. GST inhibited iNOS mRNA expression induced by LPS plus IFN-$\gamma$, whereas NaSA did not. These findings suggest that GST may exert anti-arthritic, anti-rheumatic and anti-inflammatory effect by inhibiting iNOS expression induced by LPS plus IFN-$\gamma$ at transcriptional level, whereas NaSA exert its effect by inhibiting iNOS expression at the translational or posttranslational level.

Involvement of GTP-Binding Proteins in Stage-Specific Receptor-Mediated Endocytosis of Coelomic Fluid Proteins into Oocytes of Pseudopotamilla occelata (안점의 꽃갯지렁이 난포세포로 체강액 단백질의 단계특이적 유입을 위한 GTP-Binding Protein의 개입)

  • 남현정;강화선;이양림
    • The Korean Journal of Zoology
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    • v.39 no.3
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    • pp.292-298
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    • 1996
  • Receptor-mediated endocytosis of coelomic fluid proteins (CP), yolk precursor proteins, appears to be regulated by multiple GTP-binding proteins during oogenesis of a polychaete, Pseudopotamilla occelata. Transport of 125 I-CP into the oocytes of intermediate size class, at which CP is the most actively transported, is enhanced by GTP but inhibited by GTP analogues, either GTPrS or GTP$\beta$S. The effects of GTP and GTPrS on the transport were also confirmed by tracing internalization of gold-labeled CP with transmission electron microscope. Internalization of gold-labeled CP into the yolk granules was enhanced by GTP but inhibited by GTPrS.

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An Immunocytochemical Study on Storage Proteins of Ginseng Seed - Tris Buffer Soluble Protein - (인삼 종자의 저장단백질에 관한 면역 세포화학적 연구 - Tris 완충액 가용성 단백질 -)

  • Kim, Woo-Kap
    • Applied Microscopy
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    • v.19 no.2
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    • pp.74-84
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    • 1989
  • Buffer soluble storage proteins of ginseng seed have been localized by electron microscopy using post-embedding immunocytochemical gold labelling technique. Major components of the storage proteins were revealed to be storage protein-1($SP_{1}$, MW 160,000) and storage protein-2($SP_{2}$, MW 70,000). Both of the storage proteins are glycoproteins. Anti-$SP_{1}$ and anti-$SP_{2}$ from rabbit, against $SP_1$ and $SP_2$, respectively, reacted on sections of ginseng endosperm tissue embedded in Spurr's epoxy resin. The rabbit antibodies were visualized indirectly by reaction with protein A labelled with colloidal gold. Both storage proteins were found to be accumulated together in the same protein bodies, but their relative contents are not equal.

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Development of Rapid Immune-gold Strip Kit for On-Site Diagnosis of Tomato spotted wilt virus

  • Yoon, Ju-Yeon;Choi, Gug-Seoun;Cho, In-Sook;Choi, Seung-Kook
    • Research in Plant Disease
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    • v.20 no.1
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    • pp.15-20
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    • 2014
  • A rapid, user-friendly and simple immune-chromatographic dipstick kit named 'rapid immune-gold strip' (RIGS) kit was developed in a novel single strip format to detect on-site detection of Tomato spotted wilt virus (TSWV). Immunoglobulin G (IgG) from polyclonal antisera raised in rabbits against TSWV was purified through protein-A affinity chromatography and then the purified TSWV-IgG was conjugated to colloidal gold nano-particles which served as a test line on nitrocellulose membrane. Protein A that non-specifically binds to TSWV antibody was used as a control line on the same strip. The diagnosis process with the TSWV-RIGS involves simply grinding the suspect plant sample in a bag that contains the extraction buffer and inserting the strip the bag. Results can be seen in 2-5 minutes. The flow of the complexes of gold particles coated with TSWV-IgG and a crude sap from TSWV-infected pepper, tobacco and tomato plants resulted in intensive color formed on the test lines proportional to the concentrations of TSWV. The RIGS-TSWV kit did not show any cross-reactions against other tomato-infecting viruses unrelated to TSWV. These results indicate that the TSWV-RIGS kit is highly sensitive and is not required for laboratory training and experience prior to testing. The TSWV-RIGS kit is suitable for on-site detection of suspect TSWV-infected plants as well as for laboratory diagnosis.

The Diagnostic Value of Interferon-γ Assay in Patients with Active Tuberculosis (활동성 결핵의 진단에서 혈청 인터페론 감마 측정법의 유용성)

  • Park, So Young;Park, Yong Bum;Choi, Jeong Hee;Lee, Jae Young;Kim, Jae-Seok;Mo, Eun Kyung
    • Tuberculosis and Respiratory Diseases
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    • v.66 no.1
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    • pp.13-19
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    • 2009
  • Background: The interferon-gamma assay is reported to have high sensitivity and specificity for making the diagnosis of latent tuberculosis infection. The clinical usefulness of this essay for detecting active tuberculosis has not fully defined. We evaluated the diagnostic value of the commercial interferon-gamma assay kit (QuantiFERONTB GOLD) for patients with suspected tuberculosis. Methods: From January to August 2007, we recruited 52 patients with suspected tuberculosis infection. We performed chest X-ray, sputum smear, culture, PCR and the QuantiFERON-TB GOLD test. Pleural fluid analysis and pleural biopsy were also done for the patients with pleural effusion. Results: Of the 52 patients we studied, 30 patients had a positive QuantiFERON-TB GOLD test result. 35 patients were finally diagnosed with active tuberculosis: twenty-five with a positive QuantiFERON-TB GOLD test and 10 with a negative QuantiFERON-TB GOLD test. The sensitivity of the QuantiFERON-TB GOLD test was 71.4% and the specificity was 64.7%. The positive predictive value was 0.83 and the negative predictive value was 0.50. There was no significant difference of any of the clinical and laboratory characteristics between the two groups of patients except the C-reactive protein (CRP) level. The CRP level was 29.2${\pm}$27.3 mg/dL in the pulmonary tuberculosis patients with a positive QuantiFERON-TB GOLD test and 72.9${\pm}$67.9 mg/dL in the patients with a negative QuantiFERON-TB GOLD test (p<0.05). Conclusion: The sensitivity and specificity of the QuantiFERON-TB GOLD test were inadequate for making the diagnosis of active tuberculosis. We suggest that the QuantiFERON-TB GOLD test should not be used by itself to exclude the diagnosis of active tuberculosis. The relationship of the QuantiFERON-TB GOLD test and the CRP level in patients with TB would be further investigated.

Immunosensor for Detection of Escherichia coli O157:H7 Using Imaging Ellipsometry

  • Bae Young-Min;Park Kwang-Won;Oh Byung-Keun;Choi Jeong-Woo
    • Journal of Microbiology and Biotechnology
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    • v.16 no.8
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    • pp.1169-1173
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    • 2006
  • Imaging ellipsometry (IE) for detection of binding of Escherichia coli O157:H7 (E. coli O157:H7) to an immunosensor is reported. A protein G layer, chemically bound to a self-assembled layer of 11-mercaptoundecanoic acid (11-MUA), was adopted for immobilization of monoclonal antibody against E. coli O157:H7 (Mab). The immobilization of antibody was investigated using surface plasmon resonance. To fabricate antibody spots on a gold surface, protein G solution was spotted onto the gold surface modified with an 11-MUA layer, followed by immobilizing Mab on the protein G spot. Ellipsometric images of the protein G spot, the Mab spot, and Mab spots with binding of E. coli O157:H7 in various concentrations were acquired using the IE system. The change of mean optical intensity of the Mab spots in the ellipsometric images indicated that the lowest detection limit was $10^3$CFU/ml for E. coli O157:H7. Thus, IE can be applied to an immunosensor for detection of E. coli O157:H7 as a detection method with the advantages of allowing label-free detection, high sensitivity, and operational simplicity.

Ultrastructural Change of the Bile Duct Fibroblast at Infected Rat with Clonorchis sinensis (간흡충에 감염된 실험쥐 담관 섬유모세포의 미세구조적 변화)

  • Kim, Soo-Jin;Min, Byoung-Hoon
    • Applied Microscopy
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    • v.34 no.2
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    • pp.121-130
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    • 2004
  • In this study, ultrastructural change of the bile duct fibroblast at infected rat with Clonorchis sinensis, and the distribution of lectin receptors and actin protein in cultured bile duct infected with Clonorchis sinensis. It explored using colloidal gold label complex with lectin WGA purified from wheat germ (Triticum vulgaris) and anti actin antibody purified actin (43 kDa) isolated from chicken back muscle. The lectin WGA with protein A gold complex labeled sections of the cultured fibroblast revealed gold particles specifically distributed on the multi vesicular form Golgi complex and cell surface of the fibroblast. The actin antibody with protein A gold complex labeled sections of the cultured fibroblast revealed gold particles specifically distributed on the cytoplasm of the fibroblast. Labeling of cultured fibroblast in rat bile duct infected with Clonorchis sinensis was then quantified and compared to that of cultured Fibroblast in Rat Bile duct. These results indicate that lectin WGA receptors are located in the multi vesicular form Golgi complex in the cytoplasm to the cytoplasmic process of the Rat bile duct fibroblast infected with Clonorchis sinensis. Therefore, the GlcNAc and NeuNac regions on the cell surface and cytoplasmic process appear to be functionally associated with cell-recognition and protection from other cell of the tissue, and linked with secretion and exocytosis of the fibroblst cytoplasm. GlcNAc and NeuNAc product in the multi vesicular form Golgi complex then it is transported to cell surface. Actin protein is many appears that infected fibroblast rather than normal fibroblast. The fibroblast of infected with Clonorchis sinensis are against of the physical and chemical stimulation. Then development of cytoplasmic process is relative some stimulation.