• Maxillofacial Plastic and Reconstructive Surgery
• /
• v.20 no.2
• /
• pp.173-183
• /
• 1998

$TGF-{\beta}$ is one of growth factors that may be involved in the formation of bone and cartilage. Multiple studies demonstrate that $TGF-{\beta}$ is involved in regulating cell proliferation, differentiation and matrix synthesis, events observed in frature healing. The apperance of $TGF-{\beta}$ in the fracture during healing was evaluated by immunohistologic localization of $TGF-{\beta}$ using antibody. Twenty Sprauge-Dawley strain white male rats, each weiging about 150grams were used and divided two groups. The one group, the $2{\times}2mm$ bony defect was formed in the right femur. The other group, $4{\times}2mm$ bony defect was formed in right femur. Both group were sacrificed at 3day, 1, 2, 3, 4 week and femur were harvested, paraffin sections were stained with H & E, MT stain, immunihistochemical staining with $TGF-{\beta}$ antibody and observed under light microscope. The result were as follows: 1. New bone formation and cartilagenous tissue was seen at 3day. And in the $2{\times}2mm$ bony defect group, $TGF-{\beta}$ stained the cell surounding new bone. 2. The osteoclast and trabecular was seen at 1week. $TGF-{\beta}$ stained the osteoblast and in the $2{\times}2mm$ bony defect group was stained more than $4{\times}2mm$ bony defect group. 3. The lamella bone and trabecule was seen from 3, 4week, and $TGF-{\beta}$ stained almost negative. From the above finding, we could concluded that $TGF-{\beta}$ stained the osteoblast at early stage and 1week, the peak stain was seen from 1week, and then decreased, almost negative stain was seen at 3, 4week.

• Journal of Photoscience
• /
• v.9 no.2
• /
• pp.382-384
• /
• 2002

When illuminated with strong visible light, the reaction center Dl protein of photo system II is photodamage and degraded. Reactive oxygen species and endogenous cationic radicals generated by photochemical reactions are the cause of the damage to the Dl protein. Recently we found that the photodamaged Dl protein cross-links with the surrounding polypeptides such as D2 and CP43 in photosystem II. As the cross-linking reaction is dependent on the presence of oxygen, reactive oxygen species are suggested to be involved. Among the reactive oxygen species examined, ？ OH was most effective in the formation of the cross-linked products. These results indicate that the cross-linking is mostly due to ？ OH generated at photosystem II. The cross-linking site of the Dl protein is not known. As several tyrosine residues exist at the D­E loop of the Dl protein, there is a possibility that di-Tyr is formed between the D­E loop of the Dl protein and surrounding polypeptides during the strong illumination. Therefore, we examined the formation of di-Tyr using the monoclonal antibody against di-Tyr under excess illumination of the photosystem II membranes. The results obtained here suggest that no di-Tyr is formed during the excess illumination of photosystem II.

• Journal of Microbiology and Biotechnology
• /
• v.18 no.10
• /
• pp.1695-1700
• /
• 2008

An immunosensor based on surface plasmon resonance (SPR) with enhanced performance was developed through a mixed self-assembled monolayer. A mixture of 16-mercaptohexadecanic acid (16-MHA) and 1-undecanethiol with various molar ratios was self-assembled on gold (Au) surface and the carboxylic acid groups of 16-MHA were then coordinated to Zn ions by exposing the substrate to an ethanolic solution of $Zn(NO_3)_2\cdot 6H_2O$. The antibody was immobilized on the SPR surface by exposing the functionalized substrate to the desired solution of antibody in phosphate-buffered saline (PBS) molecules. The film formation in series was confirmed by SPR and atomic force microscopy (AFM). The functionalized surface was applied to develop an SPR immunosensor for detecting human serum albumin (HSA) and the estimated detection limit (DL) was 4.27 nM. The limit value concentration can be well measured between ill and healthy conditions.

• Bulletin of the Korean Chemical Society
• /
• v.33 no.2
• /
• pp.651-656
• /
• 2012

Cationic immunoliposomes were prepared by conjugation of Fab' fragments of the recombinant humanized monoclonal antibody (HuCC49) against tumor-associated glycoprotein (TAG)-72 to sterically unilamella liposomes. The cationic immunoliposomes are composed of cationic lipid (O,O'-dimyristyl-N-lysyl aspartate, DMKD), cholesterol, and 1,2-dipalmitoyl-sn-glycero-3-phosphoethanolamine-N-[maleimide(polyethyleneglycol)$_{2000}$] (DPPE-PEG-maleimide) with a molar ratio of 0.5:0.47:0.03. Plasmid DNA was effectively condensed by addition of transferrin (Tf) during the formation of anti-TAG-72 PEG-immunolipoplexes (PILPs). These anti-TAG-72 PILPs were able to adhere to the surface of TAG-72-overexepressing LS174T human colon cancer cells more effectively than conventional liposomes, thereby facilitating gene delivery in vitro. Furthermore, intravenous administration of the anti-TAG-72 PILPs into the tumor-carrying mice exhibited efficient localization of the reporter gene in the tumor tissues.

• Journal of fish pathology
• /
• v.24 no.2
• /
• pp.47-51
• /
• 2011

The present study demonstrated lymphocystis disease virus (LCDV) propagation through cytopathic effects (CPE) formation and LCDV detection in olive flounder fin (FFN) cells by polymerase chain reaction (PCR) and fluorescent antibody technique (FAT) methods. Tissue filtrates from the cluster cells produced CPE in FFN cells, which initially cells became enlarged and gradually underwent fusion en masse. Infectivity of culture grown LCDV using the FFN cells reached $10^{2.3}$ $TCID_{50}$/ml at 4 days post infection and the highest titer was measured $10^{6.5}$ $TCID_{50}$/ml at 12 days. The viral DNA was detected in the cell culture supernatants showing CPE and the CPE cells by PCR. Antigen specific strong fluorescence reacting with monoclonal antibody against the virus revealed the presence of viral antigen in the cytoplasm of infected FFN cells. These results suggest that the FFN cell line originated from the olive flounder has a susceptibility of the LCDV.

• Journal of Oriental Neuropsychiatry
• /
• v.6 no.1
• /
• pp.1-17
• /
• 1995

In order to investigate the Anti-stress effect of Guibiondamtang in the immobilization stressed rats, the level of serum catecholamine, the change of body weight, the humoral and cellular immune response were studied. The results were as follows; 1. The decrese of the body weight was significantly inhibited in test group for Guibiondamtang, comparting to the control group. 2. The increase of the level of serum norepinephrine was significantly inhibited in test group, comparing to the control group. 3. The increase of the level of serum epinephrine was significantly inhibited in test group, comparing to the control group. 4. In the hemagglutinaton titer, the control group was decreased on the serum antibody titer but test group was inhibitory effect on the decrease of sereum antibody titer. 5. In the plaque formation test, the control and test group were not shown significant differences. 6. In the foot pad swelling respopnse, the control group was decreased on DTH response but test group was increased comparing to the normal group. 7. There was no change on the distribution of lymphocyte subset(CD4, CD8), grnulocyte and macrophage analyzed by flow cytometry.

• Journal of Microbiology and Biotechnology
• /
• v.12 no.5
• /
• pp.780-786
• /
• 2002

An immunosensor based on surface plasmon resonance (SPR) with a self-assembled protein G layer was developed for the detection of Escherichia coli O157:H7. A self-assembled protein C layer on a gold (Au) surface was fabricated by adsorbing the mixture of 11-mercaptoundecanoic acid (MUA) and hexanethiol at various molar ratios and by activating chemical binding between free amine (-$NH_2$) of protein G and 11-(MUA) using 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide hydrochloride (EDAC) in series. The formation of a self-assembled protein G layer on an Au substrate and the binding of the antibody and antigen in series were confirmed by SPR spectroscopy. The surface morphology analyses of the self-assembled protein G layer on the Au substrate, monoclonal antibody (Mab) against E. coli O157:H7 which was immobilized on protein G, and bound E. coli O157:H7 extracts on Immobilized Mab against E. coii O157:H7 were performed by atomic force microscopy (AFM). The detection limit of the SPR-based immunosensor for E. coli O157:H7 was found to be about $10^4$ cells/ml.

• Korean Journal of Veterinary Research
• /
• v.37 no.3
• /
• pp.547-554
• /
• 1997

Galectin-3 is known as an animal ${\beta}$-galactoside-binding lectin charicterized with S-type carbohydrate recognition domain. It plays a role in growth, adherence and movement of cells. It is, also, related to the cell transformation and metastasis of tumor cells. In this study, we have expressed and purified recombinant human galectin-3 (rHgalectin-3) using E coli system and asialofetuin affinity chromatography for the future development of monoclonal antibody to Hgalectin-3, which is suggested as the tumor marker for the gastric and thyroid gland cancers. Expressed protein was confirmed as the Hgalectin-3 by immunoblot with cross-reactive murine monoclonal antibody. Lectin activity and specificity of purified protein were, also, confirmed by the competitive inhibition with galectin-3 specific carbohydrate, lactose. Like physiological galectin-3, lectin activity of the molecule was not changed in nonreduced condition. Dimer formation, furthermore, was observed at high concentration of the protein even in the reduced condition, which is well known in physiological galectin-3. These results showed purified rHgalectin-3 has the same activity and molecular nature compared to the physiological galectin-3.

• BMB Reports
• /
• v.29 no.6
• /
• pp.540-544
• /
• 1996

Ferritins from germinated pumpkin seeds were isolated by ammonium sulfate precipitation (0.55 saturation), ion-exchange chromatography on DEAE-cellulose, and gel filtration chromatographies on Sephacryl S-300 and Sephadex G-100. Pumpkin ferritin contains less iron than soybean ferritin. Pumpkin ferritin cross-reacted with anti-soybean ferritin antiserum made in rabbit, and showed two distinct antibody reactive bands, both of equal intensity. The pumpkin ferritins corresponding to the two bands were separable by centrifugation in a sucrose gradient (20~50%). The molecular weights of the native pumpkin ferritins based on the estimation of sucrose gradient centrifugation, gel filtration on Sephacryl S-300 and non-denaturing polyacrylamide gel electrophoresis appeared to be: 530~580 KD (the large molecular weight pumpkin ferritin) and 330-360 KD (the small molecular weight pumpkin ferritin) The large molecular weight pumpkin ferritin contains less iron. Both pumpkin ferritins cross-reacted with anti-soybean ferritin antibody with a spur formation suggesting partial antigenic recognition.

• Journal of Life Science
• /
• v.29 no.2
• /
• pp.256-264
• /
• 2019

Lymphotoxin ${\beta}$ receptor ($LT{\beta}R$), a member of the tumor necrosis factor receptor family, plays an important role in lymphoid tissue's architecture and organogenesis. In contrast, MLCK and ROCK play critical roles in the regulation of stress fiber (SF) formation in cells. To determine whether $LT{\beta}R$ stimulation in fibroblastic reticular cells (FRCs) is involved in these signaling pathways, myosin light chain kinase inhibitor-7 (ML-7) was used to inhibit them. ML7-treated FRCs completely blocked SFs and showed retraction and shrinkage processes comparable to those observed in agonistic anti-$LT{\beta}R$ antibody-treated cells. The inhibition of ROCK activity with Y27632-induced changes in actin cytoskeleton organization and cell morphology in FRCs. Actin bundles rearranged into SFs, and phospho-myosin light chain (p-MLC) co-localized in FRCs. We checked the level of Rho-guanosine diphosphate (RhoGDP)/guanosine triphosphate (GTP) exchange activity using FRC lysate. When $LT{\beta}R$ was stimulated with agonistic anti-$LT{\beta}R$ antibodies, Rho-GDP/GTP exchange activity was markedly reduced. Regarding $LT{\beta}R$ signaling with a focus on MLCK inhibition, we showed that the phosphorylation of MLCs was reduced by $LT{\beta}R$ stimulation in FRCs. Cytoskeleton components, such as tubulin, b-actin, and phospho-ezrin proteins acting as membrane-cytoskeleton linkers, were produced in de-phosphorylation, and they reduced expression in agonistic anti-$LT{\beta}R$ antibody-treated FRCs. Collectively, the results suggested that MLCK and ROCK were simultaneously responsible for SF regulation triggered by $LT{\beta}R$ signaling in FRCs.