• 한국해양바이오학회지
• /
• 제2권2호
• /
• pp.123-125
• /
• 2007

Vibrio anguillarum is a gram-negative bacterium that causes vibriosis in approximately 80 different fish species. V. anguillarum produces several exotoxins are correlated with the pathogenesis of vibriosis. This study is focused on the composition of the outer membrane vesicle. Most of gram-negative bacteria produce outer membrane vesicle (OMV) during cell growth. OMV was formed from the outer membrane surface of cell and than released to extracellular environment. OMV consists of outer membrane lipids, outer membrane protein (OMP), LPS, and soluble periplasmic components. Also, they contain toxins, adhesions, and immunomodulatory. Many gram-negative bacteria were studied out forming OMV. In Vibrio sp., formation of OMV by electron microscopy has been reported from V. cholerae and V. parahaemolyticus. In present study, we isolated OMV from V. anguillarum and OMV protein was separated by SDS-PAGE. Magor band was sliced and analyzed by MALDI-TOF. The major protein band of 38kDa was identified as OmpU by MALDI-TOF MS analysis.

• Journal of Periodontal and Implant Science
• /
• 제38권4호
• /
• pp.679-690
• /
• 2008

Purpose: The aim of this study was to investigate the effects of EMD on demineralized root surface using human periodontal ligament cells and compare the effects of root conditioning materials(tetracycline(TCN), EDTA). Material and Methods: Dentin slices were prepared from the extracted teeth and demineralized with TCN and EDTA. Demineralized dentin slices were incubated at culture plate with 25, 50 and $100{\mu}g/ml$ concentration of EMD. Cell attachment, alkaline phosphatase activity test, protein synthesis assay and scanning electronic microscopic examination were done. Results: Cells were attached significantly higher in EMD treated group at 7 and 14 days. Cell numbers were significantly higher in EMD treated group. Alkaline phosphatase activity was significantly higher in EMD treated group at 7 and 14 days. Protein synthesis was significantly higher in EMD treated group at 7 and 14 days. Conclusion: Enamel matrix derivatives enhance the biologic activities of human periodontal ligament cells on demineralized root surface and its effects are dependent on the concentration of EMD.

• Journal of Periodontal and Implant Science
• /
• 제39권2호
• /
• pp.149-156
• /
• 2009

Purpose: Deproteinized bovine bone substitutes are commonly used in dental regenerative surgery for treatment of alveolar defects. In this study, three different bovine bone minerals - OCS-B (NIBEC, Seoul, Korea), Bio-Oss (Geistlich - Pharma, Switzerland), Osteograft/N - 300 (OGN, Dentsply Friadent Ceramed. TN, USA) - were investigated to analyze the basic characteristics of commercially available bone substitutes. Methods: Their physicochemical properties were evaluated by scanning electron microscopy, energy dispersive X-ray spectrometer (EDS), surface area analysis, and Kjeldahl protein analysis. Cell proliferation and alkaline phosphatase (ALP) activity of human osteosarcoma cells on different bovine bone minerals were evaluated. Results: Three kinds of bone substitutes displayed different surface properties. Ca/P ratio of OCS - B shown to be lower than other two bovine bone minerals in EDS analysis. Bio-Oss had wider surface area and lower amount of residual protein than OCS - B and OGN. In addition Bio - Oss was proved to have lower cell proliferation and ALP activity due to lots of residual micro particles, compared with OCS - B and OGN. Conclusions: Based on the results of this study, three bovine bone minerals that produced by similar methods appear to have different property and characteristics. It is suggested that detailed studies and quality management is needed in operations for dental use and its biological effects on new bone formation.

• Journal of Microbiology and Biotechnology
• /
• 제27권9호
• /
• pp.1609-1616
• /
• 2017

The complex roles of cell surface modification in the biofilm formation of Campylobacter jejuni, a major cause of worldwide foodborne diarrheal disease, are poorly understood. In a screen of mutants from random transposon mutagenesis, an insertional mutation in the eptC gene (cj0256) resulted in a significant decrease in C. jejuni NCTC11168 biofilm formation (<20%) on major food contact surfaces, such as polystyrene and borosilicate glass, when compared with wild-type cells (p < 0.05). In C. jejuni strain 81-176, the protein encoded by eptC modified cell surface structures, such as lipid A, the inner core of lipooligosaccharide, and the flagellar rod protein (FlgG), by attaching phosphoethanolamine. To assess the role of eptC in C. jejuni NCTC11168, adherence and motility tests were performed. In adhesion assays with glass surfaces, the eptC mutant exhibited a $0.77log\;CFU/cm^2$ decrease in adherence compared with wild-type cells during the initial 2 h of the assay (p < 0.05). These results support the hypothesis that the modification of cell surface structures by eptC affects the initial adherence in biofilm formation of C. jejuni NCTC11168. In motility tests, the eptC mutant demonstrated reduced motility when compared with wild-type cells, but wild-type cells with the transposon inserted in a gene irrelevant to biofilm formation (cj1111c) also exhibited decreased motility to a similar extent as the eptC mutant. This suggests that although eptC affects motility, it does not significantly affect biofilm formation. This study demonstrates that eptC is essential for initial adherence, and plays a significant role in the biofilm formation of C. jejuni NCTC11168.

• Parasites, Hosts and Diseases
• /
• 제54권4호
• /
• pp.385-391
• /
• 2016

The discovery and understanding of antigenic proteins are essential for development of a vaccine against malaria. In Plasmodium falciparum, Pf92 have been characterized as a merozoite surface protein, and this protein is expressed at the late schizont stage, but no study of Pv92, the orthologue of Pf92 in P. vivax, has been reported. Thus, the protein structure of Pv92 was analyzed, and the gene sequence was aligned with that of other Plasmodium spp. using bioinformatics tools. The recombinant Pv92 protein was expressed and purified using bacterial expression system and used for immunization of mice to gain the polyclonal antibody and for evaluation of antigenicity by protein array. Also, the antibody against Pv92 was used for subcellular analysis by immunofluorescence assay. The Pv92 protein has a signal peptide and a sexual stage s48/45 domain, and the cysteine residues at the N-terminal of Pv92 were completely conserved. The N-terminal of Pv92 was successfully expressed as soluble form using a bacterial expression system. The antibody raised against Pv92 recognized the parasites and completely merged with PvMSP1-19, indicating that Pv92 was localized on the merozoite surface. Evaluation of the human humoral immune response to Pv92 indicated moderate antigenicity, with 65% sensitivity and 95% specificity by protein array. Taken together, the merozoite surface localization and antigenicity of Pv92 implicate that it might be involved in attachment and invasion of a merozoite to a new host cell or immune evasion during invasion process.

• BMB Reports
• /
• 제35권1호
• /
• pp.61-66
• /
• 2002

The TNF receptor-associated factor (TRAF) family is a group of adapter proteins that link a wide variety of cell surface receptors. Including the TNF and IL-1 receptor superfamily to diverse signaling cascades, which lead to the activation of NF-${\kappa}B$ and mitogen-activated protein kinases. In addition, TRAFs interact with a variety of proteins that regulate receptor-induced cell death or survival. Thus, TRAF-mediated signals may directly induce cell survival or interfere with the death receptor-induced apoptosis.

• 한국동물학회지
• /
• 제33권4호
• /
• pp.446-453
• /
• 1990

본 실험은 레티노익산에 의해 F9 Teratocarcinoma Stem Cell의 분화를 유도하고 이분화과정에서 세포형태의 변화와 라미닌유전자의 발현을 조사 하였다. 분화되지 않은 F9 Stem Cell은 지속적으로 증식을 하며 세포간의 간격을 구분하기 어령루 뿐만 아니라 불규칙적인 모양을 하고 있으나,레티노익산과 dibutyryl cyclic AMP처리후의 분화된 F9세포는 둥글고 평평한 모양을 나타내며 세포성장은 중지되었다. Northern blot분석에 의하여 레티노익산과 cyclic AMP처리 후의 F9세포내에서 라미닌 유전자의 발현은 현저하게 증가하였다. 즉, 라미닌 B1유전자 발현은 분화과정 동안 최소한 30배, 라미닌 B2 유전자의 발현은 약 20배 증가하였다. 또한 라미닌 항체를 이용한 면역형광 분석결과는 Northern 분석결과와 일치하게 분화 후에 라미닌 단백질 합성이 크게 증가되었으며, 생성된 라미닌 단백질은 거의 세포표면에 분포된 것으로 나타났다. 이러한 결과로부터, 레티노익산에 의해 F9 Stem Cell의 분화가 유도되며 이 분화과정에서의 형태적인 변화와 진행은 라미닌의 생성과 밀접한 관련이 있다고 추측된다.

• 미생물학회지
• /
• 제30권5호
• /
• pp.333-338
• /
• 1992

Saccharomyces cerevisiae 의 mother cell과 daughter cell 의 연결 부위의 원형질막 바로 안쪽에 존재하는 10-nm filament ring 은 세포형태 형성과정에 중요한 역할을 하리라 간주되나 그 명확한 생성기작과 기능은 밝혀지지 않았다. 본연구에서는 CDC12 유전자로부터 gene fusion technique 을 이용하여 CDC12 단백질을 만들고 이로부터 항체를 형성하였다. 이항체를 이용하여 10-nm filament ring 의 생성기작과 기능에 대하여 연구하였다. 그 결과 CDC12 단백질은 cell cycle 전주기동안 항상 정이되나 bud 가 나오기 바고 직전에 bud 가 나올 부위에 polymerization 되었다가 세포질분열 바로 직후에 unpolymerization 되며 cytoskeletal element 의 일종인 actin 과는 무관하게 행동하는 건이 밝혀졌다. 이러한 10-nm filament 는 bud 가 나올 부위의 올바른 선정과 세포질 분열에 중요한 역할을 하리라 간주된다.

• BMB Reports
• /
• 제29권6호
• /
• pp.530-534
• /
• 1996

Astrocyte are the major glial cell type in the central nervous system (CNS), and analogous to macrophage, mediates the number of immune responses such as production of cytokines including tumor necrosis factor alpha ($TNF-{\alpha}$) upon activation. $TNF-{\alpha}$ has been implicated in neuroimmunological disorders through killing oligodendrocytes and thus causing demyelination. It has been previously demonstrated that mitogen-activated T cells synthesized a 26 kDa precursor form of $TNF-{\alpha}$ which is bound to the surface of a membrane, and is later secreted as a 17 kDa mature version. In order to examine whether astrocytes would produce the transmembrane form of $TNF-{\alpha}$, astrocytes were stimulated with biological stimuli and the membrane form of $TNF-{\alpha}$ was analyzed by Western blot and FACS analysis. When astrocytes are stimulated with lipopolysaccharide (LPS), $IFN-{\gamma}/LPS$, or $IFN-{\gamma}/IL-1{\beta}$, they were able to express a membrane-anchored $TNF-{\alpha}$ of approximately 26 kDa protein which was immunoreactive to an $anti-TNF-{\alpha}$ antibody, whereas unstimulated astrocytes or astrocytes treated with $IFN-{\gamma}$ or $IL-1{\beta}$ alone was not. Our FACS data were also consistent with the immunoblot analysis. Our result suggests that the membrane form of $TNF-{\alpha}$ expressed by activated astrocytes may cause local damage to oligodendrocytes by direct cell-cell contact and contribute to demyelination observed in multiple sclerosis (MS) and experimental allergic encephalomyelitis (EAE).

• Toxicological Research
• /
• 제14권3호
• /
• pp.371-377
• /
• 1998

We have cloned a soluble type human folate receptor(hFR type${\gamma}$) from human thymus cDNA library using the PCR amplification technique. To examine whether hFR type${\gamma}$ has a folate transport activity, CHO cells were transfected with the pcDNAhFR${\gamma}$ expression plasmid, and the stable cell line CHO/hFR${\gamma}$ expressing a high level of the hFR type${\gamma}$ was identified by northern and western blot analysis. The CHO/hFR${\gamma}$ cells produced a [$H^3$]folic acid binding protein in the culture medium. However, we couldn't detect any cell surface [$H^3$] folic acid binding and transport activities. The growth of the CHO/hFR${\gamma}$ cells was more rapidly inhibited than the wild type CHO cells in the low concentration folic acid media. These observations indicate that although soluble type human folate receptor can bind [$H^3$]folate, it does not involve in folate transport.