• Journal of Life Science
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• v.20 no.3
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• pp.365-373
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• 2010

To monitor newly emerged influenza virus variants and to investigate the prevalence pattern, our laboratory performed isolation of the viruses from surveillance sentinel hospitals. In the present study, we analysed influenza A/H1N1, A/H3N2, B viruses isolated in Busan during the 2006/07 and 2007/08 seasons by sequence analysis of the hemagglutinin (HA1 subunit) and neuraminidase (NA) genes. The isolates studied here were selected by the stratified random sample method from a total of 277 isolates, in which 15 were A/H1N1, 16 were A/H3N2 and 29 were B. Based on the phylogenetic tree, the HA1 gene showed that A/H1N1 isolates had a 96.7% to 97.7% homology with the A/Brisbane/59/2007, A/H3N2 isolates had a 98.4% to 99.7% homology with the A/Brisbane/10/2007, and B isolates had a 96.5% to 99.7% homology with the B/Florida/4/2006(Yamagata lineage), which are all the vaccine strains for the Northern Hemisphere in 2008~2009 season. In the case of the NA gene, A/H1N1 isolates had 97.8% to 98.5% homologies, A/H3N2 isolates had 98.9% to 99.4% homologies, and B isolates had 98.9% to 100% homologies with each vaccine strain in the 2008~2009 season, respectively. Characterization of the hemagglutinin gene revealed that amino acids at the receptor-binding site and N-linked glycosylation site were highly conserved. These results provide useful information for the control of influenza viruses in Busan and for a better understanding of vaccine strain selection.

• Microbiology and Biotechnology Letters
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• v.31 no.3
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• pp.271-276
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• 2003

Bacillus amyloliquefaciens D4, which produces a strongly fibrinolytic enzyme, was isolated from Chungkook-Jang, a traditional Korean soybean-fermented food. B. amyloliquefaciens D4 was mutated with N-methyl-N-nitro-N-nitrosoguanidine (MNNG) to yield a series of mutants with increasing levels of fibrinolytic enzyme production. After mutation, a mutant D4-7 was obtained with fibrinolytic activity about eight times stronger than the parent strain. The fibrinolytic activity of B. amyloliquefaciens D4-7, reached a maximum, when the producer was cultivated in 2% Isolated Soy Protein (ISP) broth for 48 h at $37^{\circ}C$. Compared to commercial fibrinolytic enzymes, the cell-free culture supernatant of B. amyloliquefaciens D4-7 showed stronger activity than plasmin and streptokinase. The optimum temperature and pH were $50^{\circ}C$ and 10.0 and thermostability was increased by the addition of glycerol, glucose, and NaCl.

• Journal of Life Science
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• v.20 no.2
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• pp.260-268
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• 2010

The lactate dehydrogenase (EC 1.1.1.27, LDH) isozymes in tissues from Channa argus were purified and characterized by biochemical, immunochemical and kinetic methods. The activity of LDH in skeletal muscle was the highest at 380.4 units and those in heart, eye and brain tissues were 13.4, 3,5 and 5.4 units, respectively. Citrate synthase (EC 4.1.3.7, CS) activity in heart tissue was the highest at 20.7 units. LDH/CS in skeletal muscle, heart, eye and brain tissues were 172.9, 0.6, 0.32 and 0.47. Protein concentration in skeletal muscle tissue was 14.7 mg/g and specific activities of LDH in skeletal muscle, heart, eye and brain tissues were 25.88, 0.79, 0.31 and 1.38 units/mg, respectively. Therefore, skeletal muscle tissue was anaerobic and heart tissue was aerobic. The LDH isozymes in tissues were identified by polyacrylamide gel electrophoresis, immunoprecipitation and Western blot with antiserum against $A_4$, $B_4$, and eye-specific $C_4$. LDH $A_4$, $A_3B$, $A_2B_2$. $AB_3$ and $B_4$ isozymes were detected in every tissue, $C_4$, $AC_3$, $A_2C_2$ and $A_3C$ were detected in eye tissue, and $A_3C$ was found in brain tissue. LDH $A_4$, $A_3B$, $A_2B_2$, $AB_3$, $B_4$, eye-specific $C_4$ isozymes were purified by affinity chromatography and Preparative PAGE Cells. The LDH $A_4$ isozyme was purified in the fraction from elution with $NAD^+$ containing buffer of affinity chromatography. Eye-specific $C_4$ isozyme was eluted right after $A_4$, after which $B_4$ isozyme was eluted with plain buffer. As a result, one part of molecular structures in $A_4$, $B_4$ and eye-specific $C_4$ were similar, but were different from each other in $B_4$ and $C_4$. Therefore the subunit A may be conservative in evolution, and the evolution of subunit B seems to be faster than that of subunit A. The activity of LDH $A_4$, $A_2B_2$, $B_4$, and eye-specific $C_4$ isozymes remained at 39.98, 21.28, 19.67 and 16.87% as a result of the inhibition by 10 mM of pyruvate, so the degree of inhibition was very high. The $Km^{PYR}$ values were 0.17, 0.27 and 0.133 mM in $A_4$, $B_4$ and eye-specific $C_4$ isozymes, respectively. The optimum pH of LDH $A_4$, $B_4$, eye-specific $C_4$, $A_2B_2$, $A_3B$, and $AB_3$ were pH 6.5, pH 8.5, pH 5.5, pH 6.0-6.5, pH 5.0 and pH 7.5. The $A_4$ and heterotetramer isozymes stabilized a broad range of pH. Especially, LDH activities in skeletal muscle tissue were high, resulting in a high degree of muscle activity.LDH metabolism in eye tissue seems to be converted faster from pyruvate to lactate by eye-specific $C_4$ isozyme as eye-specific $C_4$ have the highest affinity for pyruvate, and right after the conversion, oxidation of lactate was induced by $A_4$ isozyme. It was found that expression of Ldh-C, affinity to substrate and reaction time of $C_4$ isozyme were different according to the ecological environmental and feeding capturing patterns.

• Natural Product Sciences
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• v.3 no.2
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• pp.89-99
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• 1997

Aminolysis, hydrazinolysis and alkylation of 4-methoxy and 4,9-dimethoxy-6-cyano-7-thione-5-methyl-7H furo [3,2-g] [1] benzopyridine (1 a-b) yielded 7N-substituted furobenzopyridine derivatives (2 a-e or the possible isomers 3 a-e and 4 a-b), (5 a,b and 6 a,b) and the ester (8 a,b). Hydrolysis of (la) with acetic acid gave the corresponding pyridone derivatives (7). Furobenzopyridinyl-7-thioacetyl hydrazide (9 a,b) have been prepared via alkylation of furobenzopyridine thione (1 a-b) with ethyl chloroacetate followed by condensation with hydrazine hydrate. Schiff base (11) was prepared by reacting (9a) with p. N,N-dimethyl aminobenzaldehyde in boiling ethanol. Treatment of (8a) with anthranilic acid gave the corresponding 7-substituted-4H-3,1-benzoxazine-4-one (10). We found that compound (11) increased bleeding, coagulating time, the total count of white blood cells, blood glucose level (cause hyperglycemia), enzymes (GOT, GPT) activities, concentration of urea and creatinine. On the other hand it decreased red blood cells number, haemoglobin content and haematocrite value.

• IMMUNE NETWORK
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• v.8 no.1
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• pp.21-28
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• 2008

Background: A co-inhibitory molecule, B7-H4, is believed to negatively regulate T cell immunity by suppressing T cell proliferation and inhibiting cytokine production. However, the mechanism behind B7-H4-mediated tolerance remains unclear. Methods: Balb/c $(H-2^d)$ mice were fed with dendritic cell line, DC2.4 $(H-2^d)$ every day for 10 days. Meantime, mice were hydrodynamically injected with recombinant plasmid expressing B7-H4 fusion protein (B7-H4.hFc) or hFc via tail vein. One day after last feeding, mice were immunized with allogeneic B6 spleen cells. 14 days following immunization, mice were challenged with B6 spleen cells to ear back and the ear swelling was determined the next day. Subsequently, a mixed lymphocyte reaction (MLR) was also performed and cytokines profiles from the reaction were examined by sandwich ELISA. Frequency of immunosuppressive cell population was assayed with flow cytometry and mRNA for FoxP3 was determined by RT-PCR. Results: Tolerant mice given plasmid expressing B7-H4.hFc showed a significant reduction in ear swelling compared to control mice. In addition, T cells from mice given B7-H4.hFc plasmid revealed a significant hyporesponsiveness of T cells against allogeneic spleen cells and showed a significant decrease in Th1 and Th2 cytokines such as IFN-${\gamma}$, IL-5, and TNF-${\alpha}$. Interestingly, flow cytometric analysis showed that the frequency of CD4+CD25+FoxP3+ Tregs in spleen was increased in tolerant mice given recombinant B7-H4.hFc plasmid compared to control group. Conclusion: Our results demonstrate that B7-H4 synergistically potentiates oral tolerance induced by allogeneic cells by increasing the frequency of FoxP3+ CD4+CD25+ Treg and reducing Th1 and Th2 cytokine production.

• Bulletin of the Korean Chemical Society
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• v.31 no.4
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• pp.984-988
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• 2010

1-(Fluoren-2-yl)-benzo[d][1,2,3]triazoles 5a-b were synthesized starting from 2-nitrofluorene. 2-Nitrofluorenes 1a-b were reduced by catalytic hydrogenation, reacted with 2,4-dinitrofluorobenzene followed by catalytic hydrogenation to afford 2-(N-2,4-diaminophenyl)aminofluorenes 4a-b. Diazotization of 4a-b with $NaNO_2/H_2SO_4$ followed by treatment with $H_3PO_2$ gave 5a-b. Sulfonation of 5a-b yielded 7-benzotriazol-1-yl-fluorene-2-sulfonic acids 6a-b. The structures of 5b and 6b were firmly identified by X-ray crystal analysis in addition to $^1H$ NMR, $^{13}C$ NMR, and elemental analysis.

• Bulletin of the Korean Chemical Society
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• v.32 no.spc8
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• pp.3145-3148
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• 2011

• Journal of the Korean Society of Food Science and Nutrition
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• v.33 no.7
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• pp.1186-1191
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• 2004

Bacillus sp. $\beta$-mannanase was purified by DEAE-sephadex ion exchange column chromatography. The partially purified P-mannanase exhibited maximum activity at pH 6.0 and 5$0^{\circ}C$, and was stable at a pH range of 5.5 to 7.0, and at temperature between 30 to 5$0^{\circ}C$. Konjac glucomannan was hydrolyzed by the purified $\beta$-mannanase, and then hydrolysates separated by 1st activated carbon column chromatography and 2nd sephadex G-25 gel filtration. The main hydrolysates were composed of D.P 5 and 7 glucomannooligosaccharides by TLC and FACE method. To investigate the effects of guar gum glucomannooligosaccharides on the in vitro growth of B. longum, B. bifidum, B. infantis, B. adolescentis, B. animalis, and B. breve, Bifidobacterium spp. were cultivated individually on the modified-MRS medium containing carbon SOUTce such as D.P 5, and D.P 7 glucomannooligosaccharides, respectively. B. longum grew up 4.6-fold and 5.3-fold more effectively by the replacement of D.P 5 and 7 glucomannooligosaccharides as the carbon source in a comparasion of standard MRS. Also, B. breve and B. animalis slightly grew up by the treatment of D.P 5 glucomannooligosaccharide.

• Journal of the Korean Vacuum Society
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• v.10 no.1
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• pp.104-111
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• 2001

B-doped hydrogenated amorphous silicon carbide (a-SiC:H) thin films were prepared by plasma-enhanced chemical-vapor deposition in a gas mixture of $SiH_4, CH_4,\;and\; B_2H_6$. Physical and chemical properties of a-SiC:H films grown with varing the ratio of $B_2H_6/(SiH_4+CH_4)$ were characterized with various analysis methods including scanning electron microscopy (SEM), X-ray diffractometry (XRD), Raman spectroscopy, Fourier-transform infrared (FTIR) spectroscopy, secondary ion mass spectroscopy (SIMS), UV absorption CH_4spectroscopy and electrical conductivity measurements. With the B-doping concentration, the doping efficiency and the micro-crystallinity were decreased and the film became amorphous when $B_2H_6/(SiH_4{plus}CH_4)$ was over $5{\times}10^{-3}$. The addition of $B_2H_6$ gas during deposition decreased the H content in the film by lowering the quantity of Si-C-H bonds. Consequently, the optical band gap and the activation energy of a-SiC:H films were decreased with increasing the B-doping level.

• YAKHAK HOEJI
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• v.46 no.5
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• pp.313-319
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• 2002

Synthesis of 7$\beta$-[(Z)-2-(2-aminothiazol-4-yl)-2-(1-carboxy-1-methylethoxyimino)acetamido]-3-[[(3S, 5S)-5-[4-phenyl-5-(4-methylphenyl or 2-thiophenyl)-4H-l, 2, 4- triazol-3-yl]thiomethylpyrrolidin-3-yl]]thiomethyl-3-cephem-4-carboxylic acids (7a, 7b) were described. (2S, 4S)-4-acethylthio-2-[4-phenyl-5-(4-methylphenyl or 2-thiophenyl)-4 H-1, 2, 4-triazol-3-yl]thiomethyl-1-tert-butoxycarbonylpyrrolidines (4a, 4b) were prepared from trans-4-hydroxy-L-proline with (2S, 4R)-absolute configuration as starting material. 4-Phenyl-5-(4-methylphenyl or 2-thiophenyl)-4 H-l, 2, 4-triazol-3-thiols (2a, 2b) were prepared from p-toluic anhydride and 2-thiophene carboxylic acid hydrazide, respectively. p-Methoxybenzyl 7$\beta$-(Z)-2-(2-for-mamidothiazol-4-yl)-2-(1-tert-butoxycarbonylisopropylimino]acetamido-3-[[ (3S, 5S)-5-[4-phenyl-5-(4-methylphenyl or 2-thio phenyl)-4H-1, 2, 3-triazol-3-yl]thiomethyl-1- tert-butoxycarbonylpyrrolidin-3-yl]]thiomethyl-3-cephem-4-carboxylates (6a, 6b) were achieved by using p-methoxybenzyl ]7P-(Z)-2-(2-formamidothiazol-4-yl)-2-(tert-butoxycarbonylisopropylimino] acetamido-3-chloromethyl-3-cephem-4-carboxylate (5) and (2S, 4S)-4-acethylthio-2-[4-phenyl-5-(4-methyl phenyl or 2-thiophenyl)-4H-1, 2, 4-triazol-3-yl]thiomethyl-1-tert-butoxycarbonyl pyrrolidines (4a, 4b). Removal of formyl, Boc, and p-methoxybenzyl protecting groups were carried out by triflu oroacetic acid and anisole to give the target compounds.