• 한국생물공학회:학술대회논문집
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• 2000.11a
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• pp.191-193
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• 2000

A whole-cell biocatalyst was constructed by immobilizing an enzyme on the surface of the yeast Saccharomyces cerevisiae. The gene encoding Bacillus macerans cyclodextrin glucanotransferase(CGTase) was fused with the AGA2 gene encoding a small peptide disulfide-linked to the aga1, a cell wall protein of a-agglutinin. The plasmid was introduced S. cerevisiae and expressed in the medium consisting of 10g/L yeast extract, 20g/L peptone, and 20g/L galactose. The activity was detected with the formation of cyclodextrin(CD) from 10g/L soluble starch. Surface display of CGTase was also verified with the halo-test, flow cytometry, and immunofluorescence microscopy. The recombinant S. cerevisiae produced ${\alpha}-cyclodextrin$ more efficiently than the free CGTase by simultaneous fermentation and cyclization as yeast consumes glucose and maltose which are inhibitors for CD synthesis.

• Korean Journal of Pharmacognosy
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• v.49 no.1
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• pp.28-39
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• 2018

This study investigated the effect of snail extract on the growth parameters of old female rats (27 weeks). Rats were administered orally with snail extract at a dose of 100 mg/kg, 200 mg/kg, chondroitin sulfate 10 mg/kg and 0.9% saline (control) for 8 weeks. Bone mineral density (BMD) and serum concentrations of insulin-like growth factor 1 (IGF-1) and insulinlike growth factor-binding protein 3 (IGFBP-3) were significantly higher in rats exposed to snail extract for 8 weeks. MG-63 cells (human osteoblast-like cells) were treated with snail extract for 48 h. Their differentiation and proliferation was investigated with Western blot and morphological changes observed via immunofluorescence staining of ${\beta}-catenin$. Treatment with snail extract significantly increased the levels of growth factors including ${\beta}-catenin$ and IGF-1. The snail extract affected osteoblast formation. Morphological changes in MG-63 cells were observed via immunofluorescence staining. Treatment with snail extract increased the expression of ${\beta}-catenin$ in MG-63 cells. Results suggest that the treatment of MG-63 cells with snail extract increased the longitudinal growth and growth factor levels. Snail extract may be pharmacologically effective in osteogenic differentiation in vitro and represents a potential therapeutic agent for bone formation.

• The Korean Journal of Zoology
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• v.38 no.3
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• pp.382-387
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• 1995

To check the possible application of our anti-AFP monocional antibodies (MAbs) as immunodiagnostic reagents for hepatocellular carcinoma, ELISA and immunohistochemical assay were performed on the sera and liver biopsy specimens from the patients of hepatocellular carcinoma and other non-malignant hepatic disease. By non-competitive ELISA using anti-AFP MAbs, the highest incidence of AFP value was found only in the sera of hepatocellular carcinoma patients, i.e., more than 54% of patients had serum AFP levels of more than 500 ng/mi. By immunoperoxidase and indirect immunofluorescence techniques, anti-AFP MAbs were found to react with cytoplasm of hepatoceliular carcinoma cells. However immunohistochemical reactIvity to AFP in hepatocellular carcinoma cells was lower than that in non-neoplastic liver cells adjacent to the hepatocellular carcinoma. From these results with the similar findings from other studies, we suggest that AFP antigen is appropriate in the diagnosis assay (ELISA) but is not by immunohistochemical detection.

• The Korean Journal of Zoology
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• v.34 no.2
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• pp.209-216
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• 1991

To examine the effed of Insulin on neuronal differentiation, telencephalic neuroblasts from chick embryonic brains were cultured in a serum-free medium. Indirect immunofluorescence microscopic studies revealed that the spedfic protein, MAP-2, was localized in both cell bodies and neurites of developing neuroblasts. Furthermore, treatinent of increasing concentration of Insulin promoted the MAP-2 synthesis as well as the neurite outgrowth activity. Thus, the enhancement of the morphological and biochemical parameters for neuronal differentiation appears to he closely correlated, and the neurotrophic effect of insulin may play a crucial role in neuronal process formation.

• Clinical and Experimental Pediatrics
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• v.55 no.10
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• pp.371-376
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• 2012

Purpose: Puromycin aminonucleoside (PAN) specifically injures podocytes, leading to foot process effacement, actin cytoskeleton disorganization, and abnormal distribution of slit diaphragm proteins. p130Cas is a docking protein connecting F-actin fibers to the glomerular basement membrane (GBM) and adapter proteins in glomerular epithelial cells (GEpCs; podocytes). We investigated the changes in the p130Cas expression level in the PAN-induced pathological changes of podocytes in vitro. Methods: We observed changes in the p130Cas expression in cultured rat GEpCs and mouse podocytes treated with various concentrations of PAN and antioxidants, including probucol, epigallocatechin gallate (EGCG), and vitamin C. The changes in the p130Cas expression level were analyzed using confocal immunofluorescence imaging, Western blotting, and polymerase chain reaction. Results: In the immunofluorescence study, p130Cas showed a diffuse cytoplasmic distribution with accumulation at distinct sites visible as short stripes and colocalized with P-cadherin. The fluorescences of the p130Cas protein were internalized and became granular by PAN administration in a dose-dependent manner, which had been restored by antioxidants, EGCG and vitamin C. PAN also decreased the protein and mRNA expression levels of p130Cas at high doses and in a longer exposed duration, which had been also reversed by antioxidants. Conclusion: These findings suggest that PAN modulates the quantitative and distributional changes of podocyte p130Cas through oxidative stress resulting in podocyte dysfunction.

• Archives of Pharmacal Research
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• v.26 no.5
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• pp.358-366
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• 2003

Some N-p-substituted phenyl uracil-5-sulphonamide derivatives have been synthesized to be evaluated as molluscicides against Biomphalaria alexandrina snails, the intermediate host of Schistosoma mansoni. Schistosoma mansoni infected mice were treated with hemolymph obtained from pre-treated Biomphalaria alexandrina snails with the products 4a, 10a, 10b and 4b or obtained from non-treated snails. The selection of the concentration based on the predetermined dose which caused mortality of less than 50% of snails/24 h. $LC_{50}$ of compounds 4a, 10a, 10b and 4b was 50, 100, 200 and 50 ppm respectively. The result showed that immuno-stimulatory effect of treated hemolymph with compounds 4a, 10a and 4b was related to significant protective effects (44.4, 34.6 and 50.4% reduction in worm burden respectively). In addition, mean total worm burdens were significantly reduced in non treated hemolymph by 33.8%. The effect of hemolymph obtained from treated or non treated snails on S. mansoni adult worms antigens was studied by indirect immunofluorescence technique using chronic mouse sera (CMS). The results indicated that there was a strong reaction with epitopes in gut epithelium, tubercles, teigument and subtegumental musculature of untreated and treated S. mansoni adult worms antigens. Therefore, treatment of hemolymph obtained from pre-treated snails with compounds 4a, 10a, and 4b can stimulate specific immune response and induce protective effects against S. mansoni infection.

• Korean Journal of Microbiology
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• v.40 no.1
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• pp.23-28
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• 2004

Antigenic analysis of Herpes simplex type 2 virus was performed and its major antigen was localized using an immunoelectron microscopy. Antigens of 32, 43, 59 and 69 kDa were constantly expressed during the course of infection for 48 hr in the infected Vero cell. An antigen of 51 kDa was turned out to be the major one in inducing a immune response in Western-blot analysis. The 51 kDa antigen was localized on the surface of HSV-2 by immunoelectron microscopy using colloidal golds and anti-HSV 2 polyc1onal antibody. Immunofluorescence assay indicated that viral antigens were found throughout the infected cell and, especially, on the surface of the cell.

• International Journal of Oral Biology
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• v.43 no.4
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• pp.177-183
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• 2018

The objective of this study was to examine the expression pattern of Kelch-like ECH-associated protein 1 (Keap1) in the maxillary $2^{nd}$ molar germs of rats. We used the maxillary $2^{nd}$ molar germs in rats' pup at postnatal day 3 (bell stage), 6 (crown formation stage) and 9 (root formation stage). The investigation on mRNA and protein levels were done using reverse transcription - polymerase chain reaction and western blot. Localization of Keap 1 in the maxillary $2^{nd}$ molar germs were revealed through immunofluorescence staining. Keap1 from the maxillary 2nd molar germs were mostly manifested on postnatal day 3 and dramatically decreased on postnatal day 6 and 9 at mRNA and protein levels, while amelogenin and ameloblastin increased during the development of maxillary 2nd molar germs. During immunofluorescence analysis, the strong immunoreactivity against Keap1 was detected in the apical side of ameloblasts at the presecretory and secretory stages. However, Keap1 expression was hardly observed in the ameloblasts at the maturation stage. These results shows that Keap1 is strongly expressed in the presecretory and secretory ameloblasts of amelogenesis, and suggest that Keap1 may be a crucial molecule for the regulatory mechanisms tasked with the formation of enamel layer.

• Biomedical Science Letters
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• v.25 no.3
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• pp.282-287
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• 2019

Skin flap necrosis remains a major complication of reconstructive surgery. Euterpe oleracea Mart., popularly known as "acai berry" contains hydroxybenzoic acid, antioxidant polyphenolics and anthocyanins. These and other compounds within the acai berry confer anti-inflammatory and anti-oxidative effects. In this current study, we evaluated the protective effect of acai berry extracts on survival of random-pattern skin flaps in a murine model by histologic analysis. ICR mice were subjected to skin elevation surgery and orally administered acai berry extract (100 mg/kg) daily for 7 days. Tissues were stained with hematoxylin-eosin or Masson's trichrome to observe tissue integrity and collagen deposition. In addition, $TGF-{\beta}$ and VEGF was stained by immunofluorescence to determine anti-inflammatory cell infiltration and neovascularization, respectively. We found a decrease in inflammatory cell infiltration and increase in collagen deposition in the acai berry extract treated mice compared to control mice. Immunofluorescence staining reveal a higher number of $TGF-{\beta}$ positive cells and enhanced VEGF staining in the acai berry extract treated mice. The results from this study indicate that oral uptake of acai berry extract can promote healing and survival of surgical skin flaps in mice providing an augmentative therapeutic approach to enhancing skin flap survival.

• Journal of fish pathology
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• v.35 no.2
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• pp.241-246
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• 2022

White spot syndrome virus (WSSV) is a prevalent and virulent pathogen affecting cultured whiteleg shrimp (Litopenaeus vannamei) in Korea. In this study, seven monoclonal antibodies (mAbs) (10A12, 16C3, 17G4, 21G5, 22C4, 23B6 and 24G6) were produced by using purified WSSV. The reactivity of these mAbs was analysed by Western blot (WB), indirect immunofluorescence (IIF), and lateral flow immunochromatographic assay (LFIA). WB analysis demonstrated that three mAbs (17G4, 22C4, and 23B6) reacted specifically to VP28 with an approximate molecular weight of 24 kDa, mAb 16C3 reacted with approximately 17 kDa. IIF analysis demonstrated specific fluorescence signals on gill tissues of WSSV-infected shrimp, with five mAbs (10A12, 16C3, 22C4, 23B6, and 24G6), pleopods from WSSV-infected shrimp were used for LFIA, where, two mAbs (21G5 and 22C4) exhibited positive reaction. In conclusion, it can be inferred that the mAbs usage and specificity depends on the nature of assay used for diagnosis.