• International Journal of Oral Biology
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• 제35권3호
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• pp.107-111
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• 2010

Treponema denticola is a gram-negative anaerobe that can cause periodontal disease. The adhesion of this bacterium to host tissues is considered to be the primary event in the colonization and infection of a host. Fibrinogen is generally found in damaged tissues resulting from periodontitis. The binding ability of T. denticola to fibrinogen may therefore be an important virulence factor in inducing periodontal diseases. It has been reported recently that oral spirochetes can be labeled with fluorescent fatty acids and we speculated that this labeling method could be used in an oral spirochete binding assay. The binding of several different strains of T. denticola to immobilized human fibrinogen was therefore tested using the fluorescent fatty acid labeling method. In the case of immobilized fibrinogen, the T. denticola ATCC 35405 strain showed saturable binding to immobilized fibrinogen. Indeed, all four different T. denticola strains tested in this experiment, T. denticola ATCC 35405, T. denticola ATCC 33520, T. denticola ATCC 35404 and T. denticola OTK showed binding to fibrinogen. The fluorescent fatty acid labeling method thus shows utility in binding assays for T. denticola, different strains of which can generally bind to immobilized fibrinogen.

• 동의생리병리학회지
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• 제24권5호
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• pp.854-858
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• 2010

Streptococcus mutans (S. mutans) plays an important role in the information of dental plaque and it is being noticed as major causative bacteria of dental caries. In the present study, inhibitory effects of the ethanol extract of Dianthus superbus Linne (D. superbus) on the growth, acid production, adhesion and water-insoluble glucan synthesis of S. mutans were examined. The ethanol extract of D. superbus (0.5 - 4 mg/ml) significantly lowered the growth of S. mutans in a dose dependent manner. The acid production of S. mutans were inhibited by the presence of ethanol extract of D. superbus(1 - 4 mg/ml) significantly. The ethanol extract of D. superbus (0.25 - 4 mg/ml) also significantly lowered the adherence of S. mutans in a dose dependent manner. In water-insoluble glucan synthesis assay, 0.25 - 4 mg/ml of the ethanol extract of D. superbus significantly inhibited the formation of water-insoluble glucan. These results suggest that D. superbus may inhibit the caries-inducing properties of S. mutans. Further studies are necessary to clarify the active constituents of D. superbus responsible for such biomolecular activities.

• Macromolecular Research
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• 제11권6호
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• pp.488-494
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• 2003

We have fabricated gelatin-immobilized polyurethane foams (PUFG) by dipping polyurethane foam (PUF) in an aqueous solution containing gelatin and by subsequent reaction with glutaraldehyde after freeze-drying. Gelatin aqueous solutions of different concentrations were used as the dipping solutions to control the amount of immobilized gelatin. The average pore size of PUF decreased with an increase in gelatin concentration. It was found from the hepatocyte adhesion experiment that the amount of hepatocytes seeded on PUFG1, prepared by using a 1% aqueous gelatin solution, was higher than that on other PUFGs. The hepatocytes inoculated in PUFG1, were slightly aggregated as the incubation time increased. The cells inoculated in PUFG1 showed higher ammonia removal ability than those monolayer-cultured on a gelatin-immobilized polystyrene dish (PSG) after 1 and 4 days of incubation time. The inoculated cells exhibited higher albumin secretion relative to monolayer-cultured hepatocytes on PSG. Albumin secretion by hepatocytes seeded on PUFG1 was increased by the presence of serum and was further increased by both the presence of serum and cytokines. The results obtained from a 3-(3,4-dimethylthiazol-2yl)-2,5-diphenyltetrazolium bromide (MTT) assay indicated that PUFG can provide a better microenvironment for hepatocyte culture along with nutrition and metabolite transfer through the high porosity of PUF.

• Journal of Microbiology and Biotechnology
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• 제25권11호
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• pp.1871-1879
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• 2015

The complex roles of flagella in the pathogenesis of Campylobacter jejuni, a major cause of worldwide foodborne diarrheal disease, are important. Compared with the wild-type, an insertional mutation of the flgA gene (cj0769c) demonstrated significant decrease in the biofilm formation of C. jejuni NCTC11168 on major food contact surfaces, such as polystyrene, stainless steel, and borosilicate glass. The flgA mutant was completely devoid of flagella and non-motile whereas the wild-type displayed the full-length flagella and motility. In addition, the biofilm formation of the wild-type was inversely dependent on the viscosity of the media. These results support that flagellar-mediated motility plays a significant role in the biofilm formation of C. jejuni NCTC11168. Moreover, our adhesion assay suggests that it plays an important role during biofilm maturation after initial attachment. Furthermore, C. jejuni NCTC11168 wild-type formed biofilm with a net-like structure of extracellular fiber-like material, but such a structure was significantly reduced in the biofilm of the flgA mutant. It supports that the extracellular fiber-like material may play a significant role in the biofilm formation of C. jejuni. This study demonstrated that flgA is essential for flagellar biosynthesis and motility, and plays a significant role in the biofilm formation of C. jejuni NCTC11168.

• Journal of Microbiology and Biotechnology
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• 제13권5호
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• pp.731-737
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• 2003

Mucinase complex from V. parahaemolyticus ATCC 17802 was purified 6-fold with 0.4% yield by two sequential steps of Q-Sepharose and Superdex 200HR column chromatographies. Partially purified mucinase complex showed at least 8 times higher mucin-degrading activity than the culture filtrates. The mucinase complex also showed gelatin-and-casein-hydrolyzing activities, which demonstrates that the protein is a complex compound containing several proteases. The optimum pH and temperature of partially purified mucinase complex for mucin degradation was 8.0 and $35^{\circ}C$, respectively. The partially purified mucinase complex showed high cytotoxic activity on vero cells when examined by MTT assay and microscopic observations. Cytotoxicity was significantly increased in proportion to the concentration of the mucinase complex. Mouse experiments revealed that the jejunum, ileum. and large intestinal tissues were damaged by the injection of the mucinase complex. In particular, the reduction of the goblet cells in the large intestine was remarkable. Collectively, these data suggest that the mucinase complex partially purified from V. parahaemolyticus ATCC 17802 contributes to the adhesion and invasion of V. parahaemolyticus into the host intestinal tract.

• 한국재료학회:학술대회논문집
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• 한국재료학회 2011년도 춘계학술발표대회
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• pp.51.2-51.2
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• 2011

A novel electrospun nanofiber membrane was fabricated using combined poly (vinylphosphonic acid) (PVPA) and polyvinyl alcohol (PVA) intended for bone tissue engineering applications. PVPA is a proton-conducting polymer used as primer for bone implants and dental cements to prevent corrosion and brush abrasion. The phosphonate groups of PVPA have the ability to crosslink and attach itself to the hydroxyapatite surface facilitating faster integration of the biomaterial to the bone matrix. PVA was combined with PVPA to provide hydrophilicity, biocompatibility and improve its spinnability. To improve its mechanical strength, PVPA/PVA and neat PVA mixtures were combined to produce a multilayer scaffold. The physical and chemical properties of the of the fabricated matrix was investigated by SEM and TEM morphological analyses, tensile strength test, XRD, FT-IR spectra, swelling behavior and biodegradation rates, porosity and contact angle measurements. Biocompatibility was also examined in vitro by cytotoxicity and cell proliferation studies with MTT assay and cell adhesion behavior by SEM and confocal microscopy.

• 한국재료학회:학술대회논문집
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• 한국재료학회 2011년도 춘계학술발표대회
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• pp.52.1-52.1
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• 2011

In this work, the effect of the addition of bioactive glass in the biocompatibility and mechanical behavior of conventional TTCP/DCPA based bone cement were investigated. The cement was initially modified with chitosan and HPMC which cross-linked with citric acid to improved mechanical properties.The injectable bone substitutes were further modified by adding varying amounts of bioactive glass (0%, 10%, 20% and 30%) and its effects on the biocompatibility of the material were studied. Afterbio-glass powders were mixed with the optimized composition for HPMC and citric acid content,the IBS was incubated at $37^{\circ}C$ at different time intervals and showed progressive formation of HAp with increasing time. Mechanical properties like Vickers hardness and compressive strength were found to increase with the increasing amount of bioactive glass addition and that setting time was shortened. The fabricated IBS morphologies were further characterized using SEM. MTT assay was performed to check the cell cytotoxicity and cell proliferation for 1, 3 and 5 days. Cell morphology, adhesion and proliferation behavior of cell in the IBS by culturing MG-63 cells on the IBS for 20, 60 and 90 mins and 1, 3 and 5 days was also investigated. All the results showed increasing biocompatibility as the bioglass content increased. MTT results found the materials to be cytocompatible and SEM images showed that cells attached and proliferated successfully.

• BMB Reports
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• 제32권1호
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• pp.98-102
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• 1999

Since carbohydrates are major mediators in cell-to-cell adhesion and communication, the development of specific and strong binders against them could generate promising therapeutics. As the first step towards that goal, sugar molecules have to be immobilized to be used as an affinity matrix. The amino functionality in sugar is the most active nucleophile for the immobilization, if the amino group is available. An alternative and general method is to use the hydroxyl group as a direct nucleophile, but the quantitation of immobilized hydroxyl groups is not easily done. To overcome this limitation, we have developed a method to immobilize various isomers of monosaccharides with p-nitrophenyl groups to the beads by using their hydroxyl groups. It was found that the amount of immobilized sugar was independent of the structure of the sugar, but was dependent on the number of hydroxyl groups. We also developed a sensitive method to quantify the amount of immobilized sugar at the picomolar scale by utilizing commercially available glycosidases to release a sensitive reporter molecule, p-nitrophenol, and detect it by HPLC. This new technique would allow a facile quantitation method for immobilized sugar molecules, which could be used as the affinity matrix to develop strong binders against biologically important sugars.

• 대한의생명과학회지
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• 제12권3호
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• pp.255-259
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• 2006

Reactive oxygen species (ROS) is one of the main pathological factors in endothelial disorder. For example, an atherosclerosis is induced by the dysfunction of vascular endothelial cells. The dysfunction of vascular endothelial cells cascades to secrete intercellular adhesion molecule (ICAM)-l substance by ROS. Therefore, The ROS is regraded as an important factor of the injury of vascular endothelial cells and inducement of atherosclerosis. Oxygen radical scavengers playa key role to prevention of many diseases mediated by oxidative stress of ROS. In this study, the toxic effect of ROS on vascular endothelial cells and the effect of antioxidant, vitamin E on bovine pulmonary vascular endothelial cell line (BPVEC) treated with hydrogen peroxide were examined by the colorimetric assay. ROS decreased remarkably cell viability according to the dose- and time-dependent manners. In protective effect of vitamin E on BPVEC treated with hydrogen peroxide, vitamin E increased remarkably cell viability compared with control after BPVEC were treated with $15{\mu}M$ hydrogen peroxide for 6 hours. From these results, it is suggested that ROS has cytotoxicity on cultured BPVEC and oxygen radical scavenger such as vitamin E is very effective in prevention of oxidative stress-induced cytotoxicity.

• 대한의생명과학회지
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• 제16권4호
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• pp.221-227
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• 2010

To elucidate the invasion mechanism of pathogenic Naegleria fowleri, especially a receptor-ligand recognition, we investigated the in vitro cytotoxicity of pathogenic N. fowleri and nonpathogenic N. gruberi to murine macrophages, RAW 264.7, by adding four kinds of saccharides, ${\alpha}$-fucose, ${\beta}$-galactose, ${\alpha}$-D-mannopyranoside (${\alpha}$-mannose) and xylose. There was not enough of a difference in the cytotoxicity of N. fowleri treated with 10 mM of each saccharide. In particular, the cytotoxicity of N. fowleri was highly inhibited by 100 mM ${\alpha}$-mannose, which was 62.3% inhibition calculated by the analysis of lactate dehydrogenase (LDH) release assay. Although murine macrophages were not significantly destroyed by nonpathogenic N. gruberi under hematoxylin staining, the cytotoxicity of N. gruberi was inhibited from 31.5% to 14.5% (P<0.01) by 100 mM ${\alpha}$-mannose treatment. The binding of N. fowleri to macrophages was inhibited from 33% to 50% by 100 mM ${\alpha}$-mannose. Furthermore, as results of the adhesion assays which were performed to determine whether binding of Naegleria is mediated by saccharides-binding protein, the binding ability of N. fowleri as well as N. gruberi was inhibited by 100 mM ${\alpha}$-mannose.