• Journal of Dairy Science and Biotechnology
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• v.27 no.2
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• pp.7-16
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• 2009

Appenzeller cheese samples were prepared by addition of 0.5, 1.0, and 2.0% green tea (Camellia sinensis, CS) powder and control cheese. We examined various quality characteristics of the novel cheese, such as viable-cell counts, pH, water-soluble nitrogen (WSN), non-casein nitrogen (NCN), non-protein nitrogen (NPN), and catechin level during maturation for 16 weeks at $14^{\circ}C$. To develop a Korean natural cheese containing green tea powder, we also analyzed the changes in the polyacrylamide gel electrophoresis pattern, chemical composition, and sensory qualities. The viable cell counts of the samples were not significantly different. Until the $3^{rd}$ week, the pH of the CS cheese decreased with an increase in the maturation time. However, the pH gradually increased by the $12^{th}$ week, while WSN, NCN, NPN also increased. The WSN, NCN, NPN, and catechin values for the CS cheese samples were significantly higher than the values for the control cheese. The polyacrylamide gel electrophoretic pattern of caseins for the CS cheese indicated that this cheese degraded more rapidly than the control cheese did. In the sensory evaluation, cheese with 1.0% CS powder showed the highest scores in taste and appearance and good scores in flavor and texture. These results indicate that 1.0% CS is the optimal value for addition to cheese, and cheese containing 1.0% CS shows good physiological properties and reasonably high overall sensory acceptability.

• Current Research on Agriculture and Life Sciences
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• v.11
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• pp.31-41
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• 1993

The biochemical effects of ABA and $GA_3$ on rice seedling stages were investigated from two rice cultivars, Dongjinbyeo and Samgangbyeo. Some of the results obtained are as follows ; The shoot growth was significantly increased at all concentrations of $GA_3$ compared to the control but it was completely inhibited by ABA. The root length was not affected by all concentrations of $GA_3$ except for the highest concentration of $10^{-3}M$. ABA had an inhibitary effect on root growth and showed similar trend to the shoot growth. By increasing the concentration of $GA_3$, sugar content of rice seedlings was increased and starch content was decreased. On the contrary, the treatment of ABA revealed that sugar content was decreasing with higher level of starch content. Chlorophyll content was greatly reduced at the all concentrations of $GA_3$ in both cultivars tested. Generally, the responses of root activity decreased with treatment of $GA_3$ but no difference in the activity was observed in Samgangbyeo. Major fatty acids of rice seedlings were investigated as palmitic, oleic, and linoleic acid with little contents of linolenic acid and stearic acid. The contents of these fatty acids were decreased with ABA and increased with $GA_3$ treatment, respectively. A protein band of 24 kD in the electrophoresis was observed with ABA treated rice seedlings but not with $GA_3$.

• Journal of Acupuncture Research
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• v.22 no.3
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• pp.169-183
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• 2005

Objective : The purpose of this study was to investigate the anti-caner effect of cobrotoxin on the prostatic cancer cell line (PC-3).The goal of study is to ascertain whether cobrotoxin inhibits tile cell growth and cell cycle of PC-3, or the expression of relative genes and whether the regression of PC-3 cell growth is due to cell death or the expression of gene related to apoptosis. Methods : After the treatment of Pc-3 cells with cobrotoxin, we performed 형광현미경, MTT assay, Western blotting, Flow cytometry, PAGE electrophoresis and Surface plasmon resonance analysis to identify the cell viability, cell death, apoptosis, the changes of cell cycle and the related protein, Adk, MAP kinase. Results : 1. Compared with normal cell, the inhibition of cell growth reduced in proportion with the dose of cobrotoxin(0-16nM) in PC-3. 2. Cell viabilities of 0.1, 1, 4nM cobrotoxin treatment were decreased and those of 8, 16nM were decreased significantly. 3. S phase of cell cycle was decreased at the group of 1, 2, 4, 8, 16nM cobrotoxin, but M phase was increased at 0.1, 1, 2, 4, 8, 16nM cobrotoxin. 4. Cox-2 expression after cobrotoxin was peaked at 12hours and was decreased significantly after 6, 12, 24 hours. 5. The expression of Cdk4 was decreased dose-dependently at 1, 2, 4, 8nM cobrotoxin and was decreased siginificantly at 4, 8nM Cyclin D1 was decreased at 1, 2, 4, 8nM and Cycline E was not changed. Cycline B was decreased at 1, 2, 4, 8nM dose-dependently and was decreased siginificanlty at 2, 4, 8nM. 6. The expression of Akt was decreased at 1, 2, 4, 8nM dose-dependently and was decreased significantly at 2, 4, 8nM. 7. ERK was increased at 1, 2nM and decreased at 4, 8nM, p-ERK was increased at 1, 2, 4 nM, but decreased at 8nM. JNK and p-JNK were increased at 1, 4, 8 nM. p38 was increased at 2nM p-p38 was increased at lnM but decreased significantly at 2, 4, 8nM. 8. The nucli of normal cells were stained round and homogenous in DAPI staining, but those of PC-3 were stained condense and splitted. Apoptosis was increased dose-dependently at 2, 4, 8, 16nM and increased significantly at 2, 4, 8, 16nM. 9. Bax wasn`t changed at 1, 2, 4, 8nM and Bcl-2 was decreased significantly at 1, 2, 4, 8nM. Caspase 3 and 9 weren`t changed at 1, 2, 4nM but were decreased significantly at 8nM. Conclusions : These results indicate that cobrotoxin inhibits the growth of prostate Cancer cells, has anti-cancer effects by inducing apoptosis.

• Toxicological Research
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• v.8 no.2
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• pp.343-357
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• 1992

Tumor necrosis factor-${\alpha}$(TNF), a polypeptide hormone secreted primarily by activated macrophages, was originally identified on the basis of its ability to cause hemorrhagic necrosis and tumor regression in vivo. Subsequently, TNF has been shown to be an important component of the host responses to infection and cancer and may mediate the wasting syndrome known as cachexia. These systemic actions of TNF are reflected in its diverse effects on target cells in vitro. TNF initiates its diverse cellular actions by binding to specific cell surface receptors. Although TNF receptors have been identified on most of animal cells, regulation of these receptors and the mechanisms which transduce TNF receptor binding into cellular responses are not well understood. Therefore, in the present study, the mechanisms how TNF receptors are being regulated and how TNF receptor binding is being transduced into cellular responses were investigated in rat liver plasma membranes (PM) and ME-180 human cervical carcinoma cell lines. $^{125}I$-TNF bound to high ($K_d=1.51{\pm}0.35nM$)affinity receptors in rat liver PM. Solubilization of PM with 1% Triton X-100 increased both high affinity (from $0.33{\pm}0.04\;to\;1.67{\pm}0.05$ pmoles/mg protein) and low affinity (from $1.92{\pm}0.16\;to\;7.57{\pm}0.50$ pmoles/mg protein) TNF binding without affecting the affinities for TNF, suggesting the presence of a large latent pool of TNF receptors. Affinity labeling of receptors whether from PM or solubilized PM resulted in cross-linking of $^{125}I$-TNF into $M_r$ 130 kDa, 90 kDa and 66kDa complexes. Thus, the properties of the latent TNF receptors were similar to those initially accessible to TNF. To determine if exposure of latent receptors is regulated by TNF, $^{125}I$-TNF binding to control and TNF-pretreated membranes were assayed. Specific binding was increased by pretreatment with TNF (P<0.05), demonstrating that hepatic PM contains latent TNF receptors whose exposure is promoted by TNF. Homologous up-regulation of TNF receptors may, in part, be responsible for sustained hepatic responsiveness during chronic exposure to TNF. As a next step, the post-receptor events induced by TNF were examined. Although the signal transduction pathways for TNF have not been delineated clearly, the actions of many other hormones are mediated by the reversible phosphorylation of specific enzymes or target proteins. The present study demonstrated that TNF induces phosphorylation of 28 kDa protein (p28). Two dimensional soidum dodecyl sulfate-polyacrylamide gel electrophoresis(SDS-PAGE) resolved the 28kDa phosphoprotein into two isoforms having pIs of 6.2 and 6.1. The pIs and relative molecular weight of p28 were consistent with those of a previously characterized mRNA cap binding protein. mRNA cap binding proteins are a class of translation initiation factors that recognize the 7-methylguanosine cap structure found on the 5' end of eukaryotic mRNAs. In vitro, these proteins are defined by their specific elution from affinity columns composed of 7-methylguanosine 5'-triphosphate($m^7$GTP)-Sepharose. Affinity purification of mRNA cap binding proteins from control and TNF treated ME-180 cells proved that TNF rapidly stimulates phosphorylation of an mRNA cap binding protein. Phosphorylation occurred in several cell types that are important in vitro models of TNF action. The mRNA cap binding protein phosphorylated in response to TNF treatment was purifice, sequenced, and identified as the proto-oncogene product eukaryotic initiation factor-4E(eIF-4E). These data show that phosphorylation of a key component of the cellular translational machinery is a common early event in the diverse cellular actions of TNF.

• Tuberculosis and Respiratory Diseases
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• v.43 no.6
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• pp.842-851
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• 1996

Background : Rifampicin(RFP) is a key component of the antituberculous shon-course chemotherapy and the RFP-resistance is a marker of multi-drug resistant(MDR) M. tuberculosis. rpoB gene encodes the ${\beta}$-subunit of RNA polymerase of M. tuberculosis which is the target of RFP. Recent reports show that rpoB gene mutations are the cause of RFP resistance of M. tuberculosis and the main mechanism of rpoB gene mutation is point mutation. And PCR-SSCP is a rapid and easy method for detecting point mutations. So we performed PCR-SSCP of rpoB gene of M. tuberculosis and compared the result with traditional RFP sensitivity test. Method : The 27 RFP sensitive M. tuberculosis culture isolates and 25 RFP resistant isolates were evaluated. The RFP sensitivity test was done at the Korean Tuberculosis istitute. The DNA was extracted by bead beater method and was amplified with primers TR-8 and TR-9 in a 20ul PCR reaction containing 0.1ul(luCi) [${\alpha}-^{32}P$] - dCTP. After amplification, SSCP was done using non-denaturaring polyacrylamide gel electrophoresis. Then direct sequencing was done in cases of different eletrophoretic mobility compared with that of H37Rv. In 19 cases, we compared PCR-SSCP results with patient's clinical course and the results of traditional RFP sensitivity test. Results : 1) All 27 RFP sensitive M. tuberculosis isolates showed the same electrophoretic mobility compared with that of H37Rv. And all 25 RFP resistant M. tuberculosis isolates showed different electrophoretic mobility. 2) The mechanism of rpoB gene mutation of M. tuberculosis is mainly point mutation. 3) The PCR-SSCP results correlate well with traditional RFP sensitivity and patient's clinical response to antituberculous treatment. Conclusion: The PCR-SSCP of rpoB gene is a very sensitive and rapid mehod in detecting RFP- resistant M. tuberculosis.

• Journal of Life Science
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• v.32 no.2
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• pp.135-141
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• 2022

Thymosin β-4 (TB4) is a small peptide composed of 43 amino acids. To obtain sufficient biologically active mouse TB4 economically, we cloned and overexpressed this gene in an Escherichia coli system. With the isopropyl β-D-1-thiogalactopyranoside induction of the E. coli transformant, TB4 fusion protein with intein- and chitin-binding domain was successfully expressed in the soluble fraction within the E. coli cell. The TB4-intein - chitin-binding domain fusion protein was purified from the soluble fraction of E. coli cell lysate. The affinity chromatography with chitin beads and dithiothreitol-mediated intein self-cleavage reaction releases the TB4 peptide into the stripping solution. Sodium dodecyl sulphate - polyacrylamide gel electrophoresis and Western blot analyses were used to confirm that the recombinant TB4 peptide was produced with the expected size of 5 kDa. We found that the recombinant TB4 stimulated cell migration in the transwell plate chamber assay. After 18 hr of the treatment of the recombinant TB4 with 1 ng/ml concentration, the migration of the HT1080 cell was increased by 20% compared with that of the chemically synthesized TB4. The recombinant TB4 was also observed to promote the healing of a wound area in C57BL/6 mice by as high as 35% compared with that of the chemically synthesized TB4. These results suggest that the recombinant TB4 has better biological activity for cell migration and wound healing than that of the chemically synthesized TB4 peptide.

• KOREAN JOURNAL OF CROP SCIENCE
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• v.22 no.1
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• pp.135-166
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• 1977

Some biochemical features of varietal variation in seed protein and their implications for soybean breeding for high protein were pursued employing 86 soybean varieties of Korea, Japan, and the U.S.A. origins. Also, studied comparatively was the temporal pattern of protein components accumulation during seed development characteristic to the high protein variety. Seed protein content of the 86 soybean varieties varied 34.4 to 50.6%. Non-existence of variety having high content of both protein and oil, or high protein content with average oil content as well as high negative correlation between the content of protein and oil (r=-0.73$^{**}$) indicate strongly a great difficulty to breed high protein variety while conserving oil content. The total content of essential amino acids varied 32.82 to 36.63% and the total content of sulfur-containing amino acids varied 2.09 to 2.73% as tested for 12 varieties differing protein content from 40.0 to 50.6%. The content of methionine was positively correlated with the content of glutamic acid, which was the major amino acid (18.5%) in seed protein of soybean. In particular, the varieties Bongeui and Saikai ＃20 had high protein content as well as high content of sulfur-containing amino acids. The content of lysine was negatively correlated with that of isoleucine, but positively correlated with protein content. The content of alanine, valine or leucine was correlated positively with oil content. The seed protein of soybean was built with 12 to 16 components depending on variety as revealed on disc acrylamide gel electrophoresis. The 86 varieties were classified into 11 groups of characteristic electrophoretic pattern. The protein component of Rm=0.14(b) showed the greatest varietal variation among the components in their relative contents, and negative correlation with the content of the other components, while the protein component of Rm=0.06(a) had a significant, positive correlation with protein content. There was sequential phases of rapid decrease, slow increase and stay in the protein content during seed development. Shorter period and lower rate of decrease followed by longer period and higher rate of increase in protein content during seed development was of characteristic to high protein variety together with earlier and continuous development at higher rate of the protein component a. Considering the extremely low methionine content of the protein component a, breeding for high protein content may result in lower quality of soybean protein.n.