• KSBB Journal
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• v.14 no.5
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• pp.578-584
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• 1999

The lectin was purified through 0.15 M NaCl extraction, ammonium sulfate precipitation, sepharose 4B affinity chromatography and gel filtration using sephadex G-150 from the leaves of Visum album collected in Mt. Duk Yu. The final gel filtration step resulted in 11.64 folds purification with 0.14% of recovery yield. We also performed biochemical characterization of the purified Visum album lectin. HPLC analysis of lectin purified by gel filtration revealed a singel peak. The analysis of the purified lectin by SDS-PAGE showed a tetramer composed of two identical subunits with molecular weights of 32 and 30 kDa. The lectin was a glycoprotein containing 14.4% carbohydrate, which consist of glucose, fructose, arabinose and xylose, and the amino acids such as phenylalanine, lysine and tyrosine. The purified lectin agglutinated human red blood cell types with similar potency, but when tested against red blood cells from mouse, bovine, rabbit, chicken and porcine, significant difference in potency were observed. Hemaggluting activity was inhibited by D-galactose, D-mannose, D-lactose and D-raffinose, but not by D-glucose, D-glucosamine, D-mannosamine, L-fructose, D-xylose, D-arabinose, D-galacturonic acid, D-fructose, L-rhamnose and N-acetyl-D-galactosamine. The optimal pH and thermal stability of the purified lectin were pH 4.0-7.0 and 20-5$0^{\circ}C$, respectively.

• Journal of Life Science
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• v.11 no.1
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• pp.57-64
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• 2001

The cDNA encoding human NeuAc ${\alpha}$2,3Gal$\beta$ 1,3GalNAc GalNac ${\alpha}$2,6-Sialyltransferase (hST6GalNac IV) was isolated by screening of human fetal liver cDNA library with a DNA probe generated from the cDNA sequence of mouse ST6Gal NAc IV (mkST6GalNAc IV). The cDNA sequence included an open reading frame coding for 302 amino acids, and comparative analysis of this cDNA with mST6GalNAc IV showed that each sequence of the predicted coding region contains 88% and 85% identifies in nucleotide and amino acid levels, respecively. The primary structure of this enzyme suggested a putative domain structure, like that in other glycosyltransferases, consisting of a short N-terminal cytoplamic domain, a transmembrane domain and a large C-terminal active domain. This enzyme expressed in COS-7 cells echibited transferase activity toward NeuAc ${\alpha}$2,3Gal$\beta$ 1,3GalNAc, fetuin and GM1b, although the activity toward the later is very low, no significant activity being detected toward Gal${\beta}$ 1,3Gal NAc or asialofetuin, the other glycoprotein substrates tested. The $^{14}$ C-sialylated residue of fetuin sialylated by this enzyem with CMP-[$^{14}$C]NeuAc was sensitive to treatment with ${\alpha}$2,8-specific sialidase of Vibrio cholerae but resistant to treatment with ${\alpha}$2,3-specific sialidase (NaNase I), and ${\alpha}$2,3- and ${\alpha}$2,8-specific sialidase of Newcastle disease virus. These results clearly indicated that the expressed enzyme is a type of GalNAc ${\alpha}$2,6-sialyltransferase like mST6GalNAc IV, which requires sialic acid residues linked to Gal${\beta}$1,3GalNAc-residues for its activity.

• Microbiology and Biotechnology Letters
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• v.36 no.1
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• pp.72-81
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• 2008

The effect of genistein on melanin synthesis was studied using in vitro and in vivo model. Genistein inhibited melanin synthesis in cultured melan-a cells dose dependently. Tyrosinase activity was decreased by genistein treatment in melan-a cells, but genistein did not inhibit tyrosinase directly. Genistein did not affect the expression of tyrosinase in melan-a cells. Genistein inhibited the activity of ${\alpha}$-glucosidase in virtro and the glycosylation of tyrosinase in melan-a cells. The resulting unsaturated glycosylation of tyrosinase makes it unstable and disturb correct transportation. To further clarify the effect of genistein on the melanogenesis, we established UVB-induced hyperpigmentation on the shaved backs of brown guinea pigs. The animals were exposed to UVB radiation once a week for three consecutive weeks. Genistein (1 and 2%) or vehicle alone as a control were then topically applied to the hyperpigmented areas daily. Genistein showed significant lightening effect on the UVB-induced hyperpigmentation in five weeks. Depigmenting effect was prominent in 2% genistein treatment with Fontana-Masson staining. In conclusion, genistein may be a useful agent for skin whitening.

• Journal of Korean Medicine Rehabilitation
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• v.23 no.4
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• pp.9-21
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• 2013

Objectives The purpose of this study was investigating effects of Jengjengamiyijin-tang (zhengzhuanjiaweierchentang) (JGYT) on lowering lipid, antioxidation and production of inflammatory mediators being used rats fed on high oxidized fat. Methods We divided fat Sprague-Dawley rats fed on high oxidized into 4 groups. Each of 8 rats was divided into a control group and experimental groups. We fed a control group of rats a basal diet and administered normal saline (100 mg/kg, 1 time/1 day) for 4 weeks. And We fed each experimental group of rats basal diet and administered an extract of JGYT extracts (100 mg/kg, 200 mg/kg, 300 mg/kg, 1 time/1 day) for 4 weeks. At the end of the experiment, the rats were sacrificed to determine their chemical composition. We measured lipid of plasma and liver, concentration of proinflammatory cytokines, antioxidative activity and plasma tumor necrosis factor-${\alpha}$ (TNF-${\alpha}$), plasma interleukin-6 (IL-6), Apo-B, Apo-E and Leptin gene expression. Results 1. Concentration of plasma FFA, LDL-cholesterol, plasma and liver total cholesterol showed a significant decrement in JGYT groups. However, concentration of plasma HDL-cholesterol showed a significant increment in JGYT groups. 2. Concentration of plasma and liver TG, TBARS showed a significant decrement in JGYT groups. However, concentration of liver GSH-Px, SOD and CAT showed a significant increment in JGYT groups. 3. Plasma GPT activity and concentration of plasma IL-6, TNF-${\alpha}$, NO, Ceruloplasmin, ${\alpha}1$-acid glycoprotein showed a significant decrement in JGYT groups. 4. In the analysis of RT-PCR, gene expression of Apo-B and Apo-E in the JGYT groups showed a low expression than that of control group. However, the gene expression of leptin showed no difference in all the treatment groups. 5. The ratio of leptin expression per ${\beta}$-actin expression showed no significant difference among all treatment groups. However, The ratio of Apo-B and Apo-E expression per ${\beta}$-actin expression showed a significant decrement in JGYT groups. Conclusions According to this study, extract of JGYT showed a positive effect in lowering lipid, antioxidation and control of inflammatory mediators production.

• The Korean Journal of Zoology
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• v.26 no.3
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• pp.155-170
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• 1983

The primary concern has been focused on the response and adaptation of mouse fibroblast tumor cells to heat-shock in the level of membrane surface proteins, using two labeling techniques, lactoperoxidase-catalyzed iodination and galactose oxidase-sodium borohydride. Cells arrested in $G_1$ phase exhibited the highest level of LETS protein and high molecular proteins than did cells passing through $G_1/S, S, G_2$ and M, and unsynchronized cells. Confluent cells were found to show an increase in 125K proteins and a decrease in 130K and 100K proteins selectively. The adaptation processes of tumor cells after heat-shock were observed. All the proteins above 80K were reduced immediately after heat-shock, whereas 70K protein increased markedly 24 hours after heat-shock. The 70K protein and high molecular proteins returned to normal level in 48 hours. The 70K protein was found to be trypsin-sensitive and was similarly labeled by galactose-oxidase as well as by lactoperoxidase. It was, therefore, concluded that 70K protein is glycoprotein located on the surface membrane and might be the HSP 70. Possible function of heat-shock protein on the surface membrane and the relation of this protein to differential heat-sensitivity of tumor cells are discussed.

• Korean Journal of Plant Resources
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• v.26 no.5
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• pp.526-532
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• 2013

In this study, we investigated the effect of 88 marine algae extracts on melanin synthesis to develop new whitening agents. Among varieties of marine algae tested, the ethyl acetate extracts from Endarachne binghamiae (EB), Scytosiphon lomentaria, Sargassum yezoense, Ecklonia cava and Sargassum fusiforme inhibited melanin synthesis in melan-a cells. EB treatment showed the strongest inhibitory activity in melanin synthesis, compared with that of other extracts. EB-mediated inhibition of melanin synthesis appeared to be associated with inhibition of ${\alpha}$-glucosidase-dependent glycosylation of tyrosinase in melan-a cells. In addition, EB treatment did not affect mushroom tyrosinase or cell-extracted tyrosinase activity in vitro. Taken together, our findings suggest that anti-browning effect of EB on skin is mediated through regulation of ${\alpha}$-glucosidase activity and subsequent inhibition of tyrosinase activity and melanin synthesis, and further development of EB as a potential agent for skin whitening.

• Korean Journal of Veterinary Research
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• v.41 no.2
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• pp.211-218
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• 2001

The $M_r$ 63,000 glycoprotein (GP63) and lipophosphoglycan (LPG) of Leishmania donovani were evaluated as vaccine candidates against visceral leishmaniasis. Mice were immunized with liposomeencapsulated GP63 and/or LPG that were purified from the soluble extract of L. donovani promastigotes, and were challenged with virulent amastigotes. Mice immunized with GP63/LPG in liposomes plus BCG resulted in a 27.4% reduction of amastigotes in the liver compared to the control group (liposomes plus BCG), and mice immunized with liposome-GP63 plus BCG failed to induce a protective immune response against the challenge infection. Immunization of mice with GP63 fused to the Schistosoma japonicum glutathione S-transferase (GP63-GST) plus BCG also failed to elicit protective immunity. To analyze the cause of failure to induce protection, cytokine release from the spleen cells of immunized mice and Leishmania-specific serum antibodies were analyzed. Spleen cells from mice immunized with GP63-GST plus BCG that were exposed to soluble extract of L. donovani in vitro produced 10-fold greater quantities of IFN-gamma and 3-fold greater quantities of IL-5 than cells from mice receiving BCG only or saline. Western blot analysis revealed that sera from these mice had Leishmania-specific antibodies recognizing 1 to 3 antigens of L. donovani with M. W. of 60-65 kDa. Although immunization of mice with GP63-GST induced a strong Th1 response, this study indicated that GP63-GST simultaneously elicited the Th2 response of the CD4+ T-cell, which was known to abrogate the protective immune response conferred by the Th1 effector function.

• Korean Journal of Bronchoesophagology
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• v.3 no.1
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• pp.70-78
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• 1997

The development of preneoplastic and neoplastic squamous cell proliferations of body sites such as the skin, female lower genital tract, and larynx is strongly associated with specific types of human papillomaviruses (HPV). Antitumor $CD^{8+}$ cells recognize peptide antigens presented on the surface of tumor cells by major histocompatibility complex (MHC) class I molecules. The MHC class I molecule is a heterodimer composed of an integral membrane glycoprotein designated the alpha chain and a noncovalently associated, soluble protein called beta-2-microglobulin( $\beta$ -2-m). Loss of $\beta$-2-m generally eliminates antigen recognition by antitumor $CD^{8+}$ T cells. We evaluated the expression of $\beta$-2-m as a potential means of tumor escape from immune recognition and the presence of HPV DNA as a cause of laryngeal squamous cell carcinomas (SCCs). Laryngeal SCCs (n=39) were analyzed for MHC class I expression by immunohistochemistry and for presence of HPV by in situ hybridization technique. The results were as follows : 1) HPV DNA was detected in 10 (25.64%) out of 39 cases in laryngeal squamous cell carcinomas. 2) MHC class I down-regulation (heterogenous and negative expression) in HPV positive lesions was higher than HPV negative lesions. 3) The expression of MHC class I was related to cellular differentiation regardless of T-stage and nodal involvement. In conclusion, HPV was thought to be the etiological factor of SCC of larynx, and we found that the down-regulation of MHC class I was a common phenomenon In laryngeal SCC and may provide a way for tumor cells to escape from immune surveillance.

• Journal of Veterinary Science
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• v.24 no.4
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• pp.51.1-51.17
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• 2023

Background: To date, various genotypes of infectious bronchitis virus (IBV) have co-circulated and in Korea, GI-15 and GI-19 lineages were prevailing. The spike protein, particularly S1 subunit, is responsible for receptor binding, contains hypervariable regions and is also responsible for the emerging of novel variants. Objective: This study aims to investigate the putative major amino acid substitutions for the variants in GI-19. Methods: The S1 sequence data of IBV isolated from 1986 to 2021 in Korea (n = 188) were analyzed. Sequence alignments were carried out using Multiple alignment using Fast Fourier Transform of Geneious prime. The phylogenetic tree was generated using MEGA-11 (ver. 11.0.10) and Bayesian analysis was performed by BEAST v1.10.4. Selective pressure was analyzed via online server Datamonkey. Highlights and visualization of putative critical amino acid were conducted by using PyMol software (version 2.3). Results: Most (93.5%) belonged to the GI-19 lineage in Korea, and the GI-19 lineage was further divided into seven subgroups: KM91-like (Clade A and B), K40/09-like, QX-like (I-IV). Positive selection was identified at nine and six residues in S1 for KM91-like and QX-like IBVs, respectively. In addition, several positive selection sites of S1-NTD were indicated to have mutations at common locations even when new clades were generated. They were all located on the lateral surface of the quaternary structure of the S1 subunits in close proximity to the receptor-binding motif (RBM), putative RBM motif and neutralizing antigenic sites in S1. Conclusions: Our results suggest RBM surrounding sites in the S1 subunit of IBV are highly susceptible to mutation by selective pressure during evolution.

• Korean Journal of Animal Reproduction
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• v.24 no.2
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• pp.125-142
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• 2000

13). Analysis of a purified preparation of eCG revealed that its $\beta$ -subunit consists of 149 amino acids, which was confirmed by the molecular cloning of its cDNA. There seem to be at least four to six, or even as many as 11, O-glycosylation sites on the extended C-tenninal region of the eCG $\beta$-subunit. Interestingly, eCG is a unique member of this family, as it appear to be a single molecule that possesses both LH- and FSH-like activities. Using the cDNA prepared from mRNA extracted from equine placental and pituitary tissues, we cloned the cDNA of eCG $\alpha$- and $\beta$ -subunits and eFSH $\beta$ -subunit. The mRNA expression of each subunit seems to be independently regulated, which may account for differences in the quantities of $\alpha$ - and $\beta$ -subunits in the placenta and pituitary. Thus, eCG is a distinct molecule from the view points of its biological function and glycoresidue structures. Recombinant eCGs including the mutants which lack oligosaccharides will be useful tools for analyzing the structure-function relationships of gonadotropins in the horse as well as other species. Similar experiments will also clarify the proposed structure and biological functions for the glycoprotein hormones. These experimental are now possible, and hopefully a resolution of the existing controversy will be forthcoming in the near future.