• Korean Journal of Veterinary Research
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• v.29 no.1
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• pp.13-20
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• 1989

The proteins of the bovine erythrocyte membrane were analyzed by polyacrylamide gel electrophoresis in sodium dodecyl sulfate, and their relations to the slow sedimentation rate of bovine erythrocytes were investigated by treating the erythrocytes with trypsin. The erythrocyte sedimentation rates of bovine erythrocytes from Holstein and Korean native cattle were very slow compared with the human one (1/7 as slow as the human one) as reported previously. However, when human and Holstein erythrocytes were treated with trypsin (0.2 and 0.5 mg/ml) for 1 hour at ${37^{\circ}C}$, their sedimentation rates were markedly accelerated while the sedimentation rate of Korean native cattle's erythrocytes were not affected. Although the general protein profiles of the bovine erythrocyte membranes were almost similar to that of human, bovine erythrocyte membranes showed one additional protein band, called band Q in this study, which migrated electrophoretically to the mid-position between band 2 and band 3 in human erythrocyte membranes. Treatment of Holstein and human erythrocytes with trypsin caused a decrease or disapperance of the band Q from the erythrocyte membrane. Although the band Q in Korean native cattle's erythroyte membrane was decreased by trypsin treatment of the erythrocytes, the magnitude of the decrement was not so pronounced as in the case of human and Holstein erythrocytes. The glycoprotein profiles of the bovine erythrocyte membranes revealed by periodic acid-Schiff stain showed a marked difference from that of human. The PAS-1 (glycophorin) and PAS-2 (sialoglycoprotein) present in human erythrocyte membrane were almost absent from the bovine erythrocyte membranes. Instead, the bovine erythrocyte membranes showed a strong PAS-positive band near the origin of the electrophorograms, which is named as PAS-B in this study. The PAS-B band was disappered completely by the trypsin treatment of Holstein erythrocytes whereas the PAS-B band in Korean native cattle's erythrocyte membrane still remained after the trypsin treatment. The trypsin treatment of Korean native cattle's erythrocytes, however, led to the appearance of small molecular weight peptides, indicating that the high molecular weight glycoproteins were degraded by trypsin as in human and Holstein ones. These results suggest that the slow sedimentation rate of bovine erythrocytes is due in part to the presence of band Q protein fraction and PAS-B glycoprotein in the bovine erythrocytes.

• Korean Journal of Microbiology
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• v.31 no.4
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• pp.279-285
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• 1993

Alterations of the p53 gene arc among the most frequent genetic changes in human cancer and often result in increased levels of p53 protein within the malignant cells. Detection of accumulated p53 protein can be a useful prognostic tool in human cancer. In order to make polyclonal antibodies for immunohistochemical screening. human p53 gene was expressed in E. coli in the form of GST (glutathione S-transfi.:rase) fusion proteins. Two p53 gene fragments. which were N('()I small fragment encoding amino acid residues of 1-151-: and Ncol large fragment of 159-393. were subeloned into the unique BamHI site present within the pGEX-2T vector using BamHI linker and recombinant plasmids pGTNS and pGTNL were constructed. respectively. The p53 cDNA fragment (from pC53-$SN_3$,) encoding amino acid 38-145 (proline at residue 72) was amplified by polymerase chain reaction(PCR). The amplified DNA was digested with BamHI and Prull and inserted into the BamHI-Smal sites of pG EX-2T and recombinant plasmid pGTBP was constructed. After IPTG induction of these plasmids for 4 hours. fusion proteins were purified from E. coli extracts with glutathione Sepharose beads. The bound proteins were resolved by 10% SDS-polyacrylamide gel electrophoresis and the molecular weights were 54 kDa. 53 kDa and 40 kDa. respectively. Approximately one milligram of fusion proteins were purified from 1 -liter cultures.

• Journal of Life Science
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• v.24 no.7
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• pp.769-776
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• 2014

Anisomycin, also known as flagecidin, is an antibiotic produced by Streptomyces griseolus that inhibits protein synthesis by binding to the ribosomal 28S subunit. The tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) is a protein that induces apoptotic cell death. TRAIL primarily causes apoptosis in tumor cells by binding to death receptors. Many human cancer cell lines are refractory to TRAIL-induced cell death. In this study, we investigated whether anisomycin could enhance TRAIL-mediated apoptosis in TRAIL-resistant human hepatocarcinoma Hep3B cells. Treatment with anisomycin and TRAIL alone did not reduce cell viability in Hep3B cells. However, in the presence of TRAIL, the anisomycin concentration dependently reduced the cell viability. Our results indicate that anisomycin sensitizes Hep3B cells to TRAIL-mediated apoptosis and that this occurs, at least partly, via caspase activation. Interestingly, Bid knockdown by small interfering RNA significantly reduced the induction of apoptosis in combination with anisomycin and TRAIL, indicating that anisomycin effectively acts to lower the threshold at which TRAIL-mediated truncated Bid triggers the mitochondrial-mediated apoptosis program in Hep3B cells. Therefore, the use of TRAIL in combination with anisomycin might provide an effective therapeutic strategy for the safe treatment of some TRAIL-resistant cancer cells.

• Journal of Plant Biology
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• v.25 no.3
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• pp.135-151
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• 1982

This investigation was carried out to clarify the changes of pigments and soluble protein, and photosystem II activity in the leaves of barley (${SO}_2$-sensitive) and corn (${SO}_2$-resistant) seedlings induced by the ${SO}_2$ fumigation (10, 50ppm). The pH changes of the leaf extract, the content of sulfite and sulfate, the activities of catalase, peroxidase, and polyphenoloxidase were compared in the leaves of barley and corn seedlings induced by ${SO}_2$ fumigation. The results are summarized as follows: An appreciable effect of pH change of leaf extract by ${SO}_2$ fumigation was observed in barley leaves (pH 6.10 to 5.18), but only a small change occurred in corn leaves (pH 5.66 to 5.50). The same pattern of pH changes was recorded when the solution of 0.2N HCl was added to leaf extract, providing lower buffering capacity of the barley leaves than corn leaves. After 2 hours of exposure to 10 ppm ${SO}_2$, the contents of ${SO}^{2-}_3$ and ${SO}^{2-}_4$ were increased in barley leaves, while only ${SO}^{2-}_4$ increased in corn leaves. After fumigation with 10ppm ${SO}_2$ for 2 hours, barley leaves showed significant decreases in activities of catalase, to 17% peroxidase, to 58%, and polyphenoloxidase, to 88%. Corn leaves showed increases in activities of peroxidase, to 136%, and polyphenoloxidase, to 128%. Absorption spectra of pigments obtained from ${SO}_2$-fumigated leaves were gradually decreased with the fumigation time increases, but the decrease was more significant in barley leaves. Fumigation with 50ppm ${SO}_2$ for 2 hours induced the greatest decomposition in carotenoid, followed by chlorophyll a and then chlorophyll b in barley leaves. The ratio of chlorophyll a/b was decreased from 4.1 to 3.6 in barley leaves, but in corn leaves it was maintained almost a constant level(4.9-4.8). The rate of decomposition of chlorophyll and carotenoid in corn leaves was very slow than those in the barley leaves. Fumigation with 50 ppm ${SO}_2$ for 2 hous, decreased the protein content of barley leaves to 59%, and that of corn leaves to 89%, and the extent of decrease in protein content was greater than that of pigments in barley and corn leaves. The rate of DCIP9dichlorophenol indophenol) photoreduction in ${SO}_2$-fumigated leaves was decreased to 18 and 67% in barley and corn leaves, respectively. However, DCIP photoreduction was considerably recovered about 32 and 92% with the addition of DPC(diphenylcarbazide) as an exogenous electron donor in barley and corn leaves, respectively.

• Korean Journal of Community Nutrition
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• v.19 no.4
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• pp.361-371
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• 2014

Objectives: This study was performed to set easily applicable portion sizes by sex and age for children at the Community Child Centers (CCC) in Korea. Methods: Considering the age and gender specific energy level at Target Patterns for children aged 6-18 years, which were suggested as a part of the 2010 Korean Food Guidance System (KFGS), we set three meal sizes. We reclassified the recommended daily servings of Grains, Meat fish eggs beans and Vegetables group at Target Patterns into three meal sizes, and then calculated the recommended serving per meal. Each proposed amount of food per meal was calculated based on serving size of foods commonly eaten at KFGS, which was then allocated to five meal components; rice, soup stew, protein and vegetable side-dishes and Kimchi. Each proposed amount of food per meal was applied to 173 menus' recipes from CANpro 3.0 as main ingredient's amounts. We cooked the 173 menus at the medium size and measured their weights after cooking. Results: Each recommended serving per meal was 0.75, 0.9 and 1.2 for Grains; 1.2, 1.6 and 2.4 for Meat fish eggs beans; 2, 2.4 and 2.8 for Vegetables by meal sizes. Among five meal components, the ratio of small and large to medium size was 1/5 less and 1/3 more for rice and 1/3 less and 1/3 more for soup stew, protein side-dish and Kimchi, respectively. We suggested the same amount for a vegetable side-dish to encourage vegetable intake. Proper portion sizes per meal of medium were rice 190 g, soup stew 210 g (solid ingredients 60 g), protein side-dish 100 g (meat eggs beans) and 70 g (fish), vegetable side-dish 80g and Kimchi 30 g. Conclusions: Proper portion size per meal suggested in this study may be useful at the CCC where dietitians are not available and the approach could be applicable to the other types of meal services.

• Journal of Ginseng Research
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• v.41 no.3
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• pp.392-402
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• 2017

Background: Previously, we reported that Korean Red Ginseng inhibited liver fibrosis in mice and reduced the expressions of fibrogenic genes in hepatic stellate cells (HSCs). The present study was undertaken to identify the major ginsenoside responsible for reducing the numbers of HSCs and the underlying mechanism involved. Methods: Using LX-2 cells (a human immortalized HSC line) and primary activated HSCs, MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-tetrazolium bromide) assays were conducted to examine the cytotoxic effects of ginsenosides. $H_2O_2$ productions, glutathione contents, lactate dehydrogenase activities, mitochondrial membrane permeabilities, apoptotic cell subpopulations, caspase-3/-7 activities, transferase dUTP nick end labeling (TUNEL) staining, and immunoblot analysis were performed to elucidate the molecular mechanism responsible for ginsenoside-mediated cytotoxicity. Involvement of the AMP-activated protein kinase (AMPK)-related signaling pathway was examined using a chemical inhibitor and small interfering RNA (siRNA) transfection. Results and conclusion: Of the 11 ginsenosides tested, 20S-protopanaxadiol (PPD) showed the most potent cytotoxic activity in both LX-2 cells and primary activated HSCs. Oxidative stress-mediated apoptosis induced by 20S-PPD was blocked by N-acetyl-$\text\tiny L$-cysteine pretreatment. In addition, 20S-PPD concentration-dependently increased the phosphorylation of AMPK, and compound C prevented 20S-PPD-induced cytotoxicity and mitochondrial dysfunction. Moreover, 20S-PPD increased the phosphorylation of liver kinase B1 (LKB1), an upstream kinase of AMPK. Likewise, transfection of LX-2 cells with LKB1 siRNA reduced the cytotoxic effect of 20S-PPD. Thus, 20S-PPD appears to induce HSC apoptosis by activating LKB1-AMPK and to be a therapeutic candidate for the prevention or treatment of liver fibrosis.

• Journal of Life Science
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• v.22 no.12
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• pp.1614-1620
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• 2012

Transgenic rice plants with increased resistance to rice brown planthopper (Nilaparvata lugens St${\aa}$l) were generated by particle bombardment-mediated transformation of plants with a gene encoding snowdrop lectin (Galanthus nivalis agglutinin; GNA) under control of the rice Rubisco small subunit (rbcS) promoter.. A large number of transgenic rice plants containing the GNA gene were generated. The integration, expression, and inheritance of this gene in the $R_1$ and $R_2$ generations were demonstrated by Southern and western blot analyses. The plants contained one to five copies of the transgene. The GNA protein comprised approximately 0.01-2.0% of total soluble protein in the $R_1$ and $R_2$ transgenic plants. Insect bioassays and feeding studies showed that the GNA protein expressed in the $R_2$ transgenic rice plants reduced the survival of brown planthoppers. The introduction of GNA into rice plants therefore can help to control insect pests.

• Korean Journal of Microbiology
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• v.44 no.4
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• pp.271-276
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• 2008

Ribosome stalling by nascent sticky peptide has been reported in several organisms across the kingdom. To test whether small subunit (SSU) rRNA is involved in this phenomenon, we developed a genetic system that utilized the specialized ribosome system to isolate SSU rRNA mutants that enable ribosomes to bypass the SecM-derived sticky peptide in protein synthesis. In this system, CAT-SecM mRNA, which encodes CAT protein containing the sticky peptide derived from SecM, is only translated by specialized ribosomes. These ribosomes were shown to transiently stall on CAT-SecM mRNA followed by the synthesis of the sticky peptide. Expression of specialized ribosomes resulted in the decreased steady-state level of CAT-SecM mRNA, which is consistent with a notion that ribosome stalling induces mRNA degradation. Isolation and characterization of SSU rRNA mutations using this genetic system that are sufficient to circumvent ribosome stalling induced by the SecM-derived sticky peptide will provide evidence of SSU rRNA function in mRNA cleavage.

• KOREAN JOURNAL OF CROP SCIENCE
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• v.51 no.1
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• pp.81-88
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• 2006

The 117 soybean cultivars were collected from nine provinces in Korea, and various seed quality traits along with isoflavone contents were evaluated to elucidate their relationship. The 100-seed weight of the black soybean (31.2 g) was significantly higher (p<0.05) than yellow soybeans (28.6 g). The composition of genistein, daidzein, and glycitein accounted for 75.8, 22.8, and 1.4 % of total isoflavone in yellow soybean cultivars, while their compositions in black soybeans were 58.5, 39.7, and 1.8%, respectively. The mean contents of total isoflavone in yellow and black soybean were $l,561.6{\mu}g\;g^{-1}\;and\;l,018.3{\mu}g\;g^{-1}$. The isofalvone content showed significant variation among cultivars when classified by the seed size. In the yellow soybeans, total isoflavone content was higher in small size soybean cultivars $(1,776.0{\mu}g\;g^{-1})$ and medium size soybean cultivars $(1,714.3{\mu}g\;g^{-1})$ compared to large size ones $(1,518.5{\mu}g\;g^{-1})$. Genistein content was proved as the major factor determining the relationship between isoflavone content and 100-seed weights (r =-0.206*). Daidzein and glycitein, however, showed no significant relationship with the 100-seed weights. Isoflavone content was not significantly correlated with color parameters L (lightness) and a (redness) values, but color parameter b (yellowness) was positively correlated with glycitein (r=0.264*) in the yellow soybeans, while its negative correlation between daidzein (r=-0.245*) and total isoflavone (r=-0.256*) were observed in black soybeans. However, these findings suggested that the seed color value may not serve as an effective parameter for estimating the isoflavone intensity of the soybeans. Variation of protein and lipid contents between yellow soybeans (n=58) and black soybeans (n=59) was relatively stable, however, protein and lipid contents have no significant relationship with isoflavone content.

• Microbiology and Biotechnology Letters
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• v.40 no.1
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• pp.50-56
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• 2012

The Wnt/${\beta}$-catenin signaling pathway regulates diverse developmental processes and adult tissue homeostasis. Inappropriate regulation of this pathway has been associated with human diseases, such as cancers, osteoporosis, and Alzheimer's disease. Using a cell-based chemical screening with natural compounds, we discovered silybin, a plant flavonoid isolated from the Silybum marianum, which activated the Wnt/${\beta}$-catenin signaling pathway in a synergy with Wnt3a-conditioned medium (Wnt3a-CM). In the presence of Wnt3a-CM, silybin up-regulated ${\beta}$-catenin response transcription (CRT) in HEK293-FL reporter cells and 3T3-L1 preadipocytes through stabilization of intracellular ${\beta}$-catenin protein. Silybin and Wnt3a-CM synergistically reduced expression of important adipocyte marker genes including peroxisome-proliferator-activated $receptor{\gamma}$ ($PPAR{\gamma}$) and CAATT enhancer-binding protein ${\alpha}$ (C/$EBP{\alpha}$) in 3T3-L1 preadipocytes, accompanied by the activation of Wnt/${\beta}$-catenin signaling pathway. Taken together, our findings indicate that silybin is a small-molecule synergist of the Wnt/${\beta}$-catenin signaling pathway and can be used as a controllable reagent for investigating biological processes that involve the Wnt/${\beta}$-catenin signaling pathway.