• 한국잠사곤충학회지
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• 제40권2호
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• pp.111-116
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• 1998

To construct transformed Bm5 cells, Autographa californica nuclear polyhedrosis virus (AcNPV)IE1 gene, an immediate early viral gene was firstly used in this study. AcNPV IE1 gene, which shares on 95.3% uncleotide sequence homology with Bombyx mori nuclear polyhedrosis virus (BmNPV) IE1 gene, was isolated and cloned into pBluescript. Neomycin gene from pKO-neo was inserted under the control of the IE1 promoter to yield pAcIE1-neo. The plasmid pAcIE1-neo was transfected into Bm5 or Sf9 cells, and neomycin-resistant cells were selected in TC100 medium containing 10% fetal bovine serum (FBS) and 1 mg/$m\ell$ G418 for two weeks. Individual clones were picked and each was amplified for further characterization. The genomic DNA from neomycin-resistnt cells was isolated and characterized by PCR using AcNPV IE1 gene-specific primers and by Southern blot analysis using neomycin gene probe. We concluded that AcNPV IE1 gene was functional in B. moridrived Bm5 cells as well as Spodaptera frugiperda-derived Sf9 cells to produce stably-transformed insect cells.

• 한국응용약물학회:학술대회논문집
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• 한국응용약물학회 1995년도 춘계학술대회
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• pp.86-86
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• 1995

The analysis of membrane receptors for hormones and neurotransmitters has progressed considerably by pharmacological and biochemical means and more recently by the use of specific anti-receptor antibodies. A 14-mer peptide (from Phe102 to Leu115 of ${\beta}$2-adrenergic receptor) was synthesized and this peptide was coupled to carrier protein Keyhole Limpet Hemocyanin(KLH) by glutaraldehyde method. A 0.5mg of KLH-coupled peptide was emulsified with equal volume of complete Freund's adjuvant and injected via popliteal lymph node to each of the three Newzealnd White rabbits. Booster injections were repeated at 4 weeks interval for three times with incomplete Freund's adjuvants. One week after the final injection, serum was prepared from ear artery. Nonspecific immunoglobulins were removed by passing the serum through KLH-Sepharose 6B affinity matrix and further by incubation with bovine lung aceton powder. The titer of the antibody for synthetic peptide which was determined by enzyme linked immunosorbent assay(ELISA) was about l/l,000. The antibody produced in this study revealed 67kDa protein band in the western blot of partially purified guinea pig lung ${\beta}$-adrenergic receptor preparation. The antibody inhibited ${\beta}$-adrenergic antaginist [3H] Dihydroalprenolol binding to soluble ${\beta}$-adrenergic receptor by 25% while control sera did not show any inhibitory effects, The result of this study suggests that the peptide sequence selected in this study may play some important roles in adrenergic receptor-ligand interaction.

• 대한지역사회영양학회지
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• 제5권3호
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• pp.432-443
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• 2000

Smoking can increase oxidative stress and thereby change the antioxidant defense system in the body. Supplementation of antioxidant vitamins might improve antioxidant status in the body. The purpose of this study was to evaluate the effectiveness of vitamin C supplementation and smoking cessation education on changes of antioxidant status and psychosocial factors related to smoking. To obtain above purposes, we investigated the effectiveness of intervention for male adolescent smokers were evalnate by assessing changes in dietary intakes, plasma antioxidant vitamin concentration, and psychosocial factors related to smoking after program completion. Subjects, male adolescent smokers, were assigned into four groups : Control group(19 students), Educ. group(19 students), Vit. C supple. group(19 students), and Educ.+Vit. C suppl. group(19 students). The Educ. group and Educ.+Vit. C suppl. group received nutrition and smoking cessation education once a week for 5 weeks. Vit. C suppl. group and Educ.+Vit. C suppl. group received 500 mg per day of ascorbic acid for 35 days. All data were collected before and after intervention. Vit. B$_2$and Vit. C intakes of all groups were increased, but the only Ca intake was increased in the Educ. group. Plasma Vit. C concentration and Ratio(plasma Vit. C/Vit. C intakes) were increased in the Vit. C suppl. group and Educ.+Vit. C suppl. group, and the Vit. C deficiency status of these groups(Vit. C suppl. group and Educ.+Vit. C suppl. group) disappeared. Showing the effects of Vit. C supplementation, plasma $\alpha$-tocopherol was increased in the Educ. and Educ.+Vit. C suppl. group, and especially high increases were seen in the Educ.+Vit. C suppl. group. Psychosocial factors related to smoking changed after the education a little. This intervention program had an impact on nutrition intakes, plasma antioxidant vitamins, and some beliefs related to smoking in male adolescent students. Various programs of nutrition and smoking cessation education and vitamin supplementation for quitting smoking must be implemented for adolescent smokers, and further studies are needed regarding sorts and amount of antioxidant nutrients and supplementation periods.

• 대한한의학방제학회지
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• 제5권1호
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• pp.147-168
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• 1997

Tish study was carried out to evaluate the possible therapeutic or antitumoral effects of Takrisodokyeum extract against tumor, and immunomodulatory effect. Some kinds of tumor were induced by the typical application of 3-methylcholanthrene (MCA) or by the implantation(s.c) of malignant tumor cells such as leukemia cells(3LL cells) or sarcoma cells(S-I80 and Fas II cells). Treatment of the Takrisodokyeum water-extract(daily 1mg mouse, i.p.) was continued for 7 days prior to tumor induction and after that the treatment was lasted for 15 days. Against squamous cell carcinoma induced by MCA, Takrisodokyeum decreased not only the frequency of tumor production but also the number and the weight of tumors per tumor bearing mice (TBM). Takrisodokyeum also significantly suppressed the development of 3LLcell and S-180 cell by frequency and their size, and some developed tumors were regressed by the continuous treatment of Takrisodokyeum extract into TBM. However, when tumor was induced by FsaII cell-implantation, the growth of implanted cells in mice was delayed by the water extract of Takrisodokyeum until day 7 and then rapid growth ensued. In vitro, treatment of Takrisodokyeum extract had no effect on the growth of some kind of cell lines such as FsaII, A-131 strain but significantly inhibited the proliferation of 3LL, S-180 cells. Takrisodokyeum also stimulated the migrative ability of leucocyte, the MIF and IL 2-production of T lymphocytes, but not IL 6 production of B cells. Takrisodokyeum enhanced Arthus reaction and DTH to sheep erythrocytes, and NK cells activities. These results demonstrated that Takrisodokyeum extract different results according to the type of tumor cells. And these results also suggested that antitumor effect of Takrisodokyeum might be chiefly due to nonspecific enhancement of NK cell activities and cell-mediated immune responses.

• Journal of the Korean Association of Oral and Maxillofacial Surgeons
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• 제34권1호
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• pp.59-70
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• 2008

Purpose: We evaluated the therapeutic effect of AEE788, a dual inhibitor of epidermal growth factor (EGF) and vascular endothelial growth factor (VEGF) receptor tyrosine kinases on human salivary adenoid cystic carcinoma (ACC) cells growing in nude mice. Experimental Design: We examined the effects of AEE788 on salivary ACC cell growth and apoptosis. To determine the in vivo effects of AEE788, nude mice with orthotopic parotid tumors were randomized to receive oral AEE788 (50 mg/kg) three times per week, injected paclitaxel ($200{\mu}g$) once per week, AEE788 plus paclitaxel, or placebo. Mechanisms of in vivo AEE788 activity were determined by immunohistochemical analysis. Results: Treatment of salivary ACC cells with AEE788 led to growth inhibition and induction of apoptosis. AEE788 inhibited tumor growth and prevented lung metastasis in nude mice. Furthermore, AEE788 potentiated growth inhibition and apoptosis of ACC tumor cells mediated by paclitaxel. Tumors of mice treated with AEE788 and AEE788 plus paclitaxel exhibited down-regulation of activated EGFR and its downstream mediators (Akt and MAPK), increased tumor and endothelial cell apoptosis, and decreased microvessel den-sity, which correlated with a decrease in the level of MMP-9, MMP-2 and bFGF expression and a decrease in the incidence of vascular metastasis. Conclusions: These data show that tumor-associated endothelial cells are important in the process of tumor-metastasis. And VEGFR can be a molecular target for therapy of metastatic lung lesion of salivary ACC.

• 셀메드
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• 제6권2호
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• pp.12.1-12.7
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• 2016

Cocos nucifera (C. nucifera) oil is indigenously used to treat cardiovascular diseases. However, coconut husk fibre (which is rich in polyphenols) has not been screened for this property. Based on the ethnomedicinal use of polyphenols in treating cardiovascular diseases, this study was carried out to evaluate the effects of polyphenols of C. nucifera husk fibre on selected cardiovascular disease indices in mice. Fifty adult male Swiss albino mice were assigned randomly into five groups (A-E). Mice in groups B, C, D and E were administered 31.25, 62.5, 125, and 250 mg/kg body weight polyphenols of ethyl acetate extract of C. nucifera husk fibre respectively while the control group (A) mice received 5% DMSO for seven days. The mice were sacrificed twenty four hours after the last administration of polyphenols. Heart and plasma lactate dehydrogenase (LDH), alanine aminotransferase (ALT), aspartate aminotransferase (AST) and alkaline phosphatase (ALP) activities and plasma lipid profile were determined. Results revealed significant reduction (^*p< 0.05) in plasma levels of total cholesterol and LDL-cholesterol with no significant change (^*p> 0.05) in HDL-cholesterol, triglyceride and VLDL levels in the plasma at all doses of polyphenols administered compared to controls. There was significant reduction (^*p< 0.05) in the activities of heart AST and LDH while plasma ALT, AST, and ALP activities were not significantly altered (^*p> 0.05) at all doses of polyphenols administered compared to controls. These results suggest that the polyphenols of C. nucifera husk fibre possess cardio-protective properties and also indicate their possible use in the treatment of cardiovascular diseases.

• 한국임상수의학회지
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• 제17권2호
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• pp.403-410
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• 2000

The Ultrasonographic method alas been widely applied to evaluating gastric motility with safety and reproducibility ill human medicine but few reference to its use in veterinary medicine is appeared. Therefore, in this study, the gastric motility was evaluated with ultrasonography by the cri-terion of mean cycle lime, short and lony axis and the area of pyloric antrum in dogs, fed with liquid of semisolid meals. Furthermore, the animals were evaluated for the effect of metoclopramide on the motility of pyloric antrum. Healthy 5 mongrel male dogs were fed with either 400 ml of milk a: a liquid meal or a mixed meal of 200 ml of milk with two pieces of bread as a semisolid meal. Mean cycle time of pyloric antrum of dogs was significantly delayed after feeding either of liquid and , semi- solid meals(P<0.05), alls it was returned to the fasting state at 60 min. after feeding of liquid meal and 160 min. after feeding of semisolid meal. Mean area of pyloric antrum of dogs was gradually decreased and was returned to the lasting state at 80 min. in doss fed liquid meal. but 1600 min. in dog\ulcorner fed semisolid meal. The administration of metoclopramide (1.0 mg/kg of of B.W.) accelerated the mean cycle time of pyloric antrum from 20 mill. to 60 min. after feeding of liquid meal and from 40 min. to 120 min. after feeding of semisolid meal. From this study, the ultrasonography was confirmed as a valuable diagnostic method leer evaluating the gastric motility and gastric area in dogs. It is non-invasive, safe and reproducible, and provides a method for the study of the effect of drugs and diseases states on gastric motility.

• 한국식품조리과학회지
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• 제33권1호
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• pp.45-53
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• 2017

Purpose: Physicochemical and microbiological qualities were investigated to compare quality characteristics of traditional with commercial soy sauce (Ganjang). Methods: Nineteen traditional products were collected from six provinces and three commercial products were purchased in domestic markets. The proximate composition, inorganic substance contents, viable bacteria, and chromaticity of the soy sauces were measured. Results: Although concentrations of crude fat and protein were not significantly different between traditional and commercial Ganjang, the moisture concentration of commercial soy sauce was significantly higher than in traditional Ganjang (p<0.05). However, the amount of ash in commercial soy sauce was significantly lower than in traditional Ganjang (p<0.05). Total nitrogen concentrations of traditional and commercial Ganjang were 0.50-1.59% and 0.86-1.26%, respectively. Concentrations of Na, Mg, K, Ca, Li, B, Fe, and Sr in traditional Ganjang were significantly higher than in the commercial products (p<0.05). The number of total bacteria in traditional and commercial Ganjang were $3.3{\times}10^1-6.4{\times}10^7CFU/mL$ and $5.5{\times}10^1-2.0{\times}10^3CFU/mL$, respectively. Bacillus cereus were below 10,000 CFU/mL in all samples, and Staphylococcus aureus was not detected. Fungi was not detected in 13 samples of traditional Ganjang and the three samples of commercial soy sauce. Although lightness, redness, and yellowness were not significantly different among the Ganjang, G10 was had the highest values (p<0.05). Conclusion: This research provided information about the quality characteristics of traditional and commercial Ganjang.

• 한국미생물생명공학회:학술대회논문집
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• 한국미생물생명공학회 2004년도 Annual Meeting BioExibition International Symposium
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• pp.273-277
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• 2004

$\alpha$-L-Arabinofuranosidase ($\alpha$-L-AFase, EC 3.2.1.55) was isolated from hyperthermophilic microorganism, Thermotoga maritima. The open reading frame (ORF) of $\alpha$-L-AFase gene is 1,455 bp long and encodes 484 amino acid residues with a molecular weight of 55,265 Da. The ORF of $\alpha$-L-AFase gene was introduced into the E. coli expression vector, $_p/RSET-B, and overexpressed in E. coli BL21. The purified recombinant $\alpha$-L-AFase showed the highest activity at 10$0^{\circ}C$ and pH 5.5. The purified enzyme appeared to have no metal cofactor requirement. The Km and specific activity values of the recombinant enzyme were 0.99 mM and 1,200 U/mg on p-nitrophenyl-$\alpha$-L-arabinofuranoside. It released only L-arabinose from sugar beet arabinan, sugar beet debranched arabinan and oat spelts arabinoxylan but had no activity onarabinogalactan and gum arabic. This result suggests that L-arabinose could be produced from natural polysaccharides using this enzyme. Mutant enzymes which Glu26, Glu172 and Glu281 residues were replaced to alanine, aspartic acid or glutamine caused Kcat to decrease by a factor of between 10$^3$ and 10$^4$. Glu172 and Glu281 residues of $\alpha$-L-AFase are seemed to be the acid/base and nucleophile in catalytic reaction, respectively, and Glu26 is supposed to playa key role in substrate binding.ng.

• Journal of Microbiology and Biotechnology
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• 제24권7호
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• pp.998-1003
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• 2014

Aspartate aminotransferase (AST; E.C. 2.6.1.1), a vitamin B6-dependent enzyme, preferentially promotes the mutual transformation of aspartate and ${\alpha}$-ketoglutarate to oxaloacetate and glutamate. It plays a key role in amino acid metabolism and has been widely recommended as a biomarker of liver and heart damage. Our study aimed to evaluate the extensive preparation of AST and its application in quality control in clinical laboratories. We describe a scheme to express and purify the 6His-AST fusion protein. An optimized sequence coding AST was synthesized and transformed into Escherichia coli BL21 (DE3) strain for protein expression. Ideally, the fusion protein has a volumetric productivity achieving 900 mg/l cultures. After affinity chromatography, the enzyme activity of purified AST reached 150,000 U/L. Commutability assessment between the engineered AST and standard AST from Roche suggested that the engineered AST was the better candidate for the reference material. Moreover, the AST showed high stability during long-term storage at $-20^{\circ}C$. In conclusion, the highly soluble 6His-tagged AST can become a convenient tool for supplying a much better and cheaper standard or reference material for the clinical laboratory.