• Korean Journal of Veterinary Research
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• v.37 no.3
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• pp.619-627
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• 1997

It is known that 2-methyl-1,4-naphtoquinone(menadione, MD) induces hepatotoxicities both in vivo and in vitro. These toxic effects are believed to result from oxidative damages to hepatocytes by "active oxygen" species via one-electron reduction of the naphtoquinone. The ginsenoside(GS) is a complex mixture of individual ginsenosides which is known to produce a range of effects on the cardiovascular and central nervous systems. In particular, GS has an antioxidant effect. In this experiment we studied the effect of GS from red panax ginseng(red ginseng total saponin, RGTS) on free radical-induced liver injuries by MD. Administration of MD($150{\mu}M$) caused an increase in aspartate aminotransferase(AST) activities and lipid peroxidation, decrease in alkaline phosphatase(ALP) activities and total bilirubin levels in blood, caused depletion of GSH and changes of antioxidant enzyme(superoxide dismutase, catalase) activities are shown in liver tissue. Administration of RGTS restored the AST levels that increased by MD, but catalase showed no significant changes. RGTS also had an effect of restoring the GSH level and had some synergistic effects with SOD. These data suggest that RGTS may have some protective effects on liver injury which is related with the oxygen free radical.

• The Journal of the Korean Society for Microbiology
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• v.21 no.2
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• pp.171-179
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• 1986

The HLA class II region encodes a series of polymorphic glycoproteins that form cell surface heterodimers each consisting of one $\alpha$ and one $\beta$ chain. Thess class II molecules are encoded by genes clustered within three loci. DP, DQ, and DR are functfonally implicated as regulatory signals in intercellular communication during the immune resposes. The phenotypic hallmark of the HLA complex is a high degree of structural and functional polymorphism. Detailed analysis. of such polymorphisms should aid in understanding the molecular basis for associations between HLA and diseases. We have used techniques of restriction enzyme fragment analysis by Southern blotting to investigate polymorphisms associated with DQ $\beta$ class II genes on haplotypes expressing the HLA-DR4 and -DQw3 specificities. The endonucleases Hind III and Bam HI were used to identify a specific DQ $\beta$ genomic polymorphism that precisely corrresponds with the reactivity of a monoclonal antibody A-10-83, previously shown to define a serologic split of DQw3. This study identifies two allelic DQ va. riants. DQw3.1 and DQw3.2. We used these specific genotypic markers to investigate the genomic basis of the association of DR4 with insulin-dependent diabetes mellitus(IDDM) and seropositive juvenile rheumatoid arthritis(JRA). The DR4 positive IDDM demonstrate the predominant expression of DQw3.2 and the very rare expression of DQw3.l. However, in haplotype matched siblings from two IDDM families, all of the DR4 positive siblings display a IDDM-associated DQw3.2 allele. Thus, both affected and healthy individuals can carry the same haplotypes and genomic markers, demonstrating that thess specific allelic variants are genetic elements that indicate a increased risk of IDDM but are not in fact disease specific. We contrasted this result with a similar analysis of patients with another DR4-associated disease, JRA. In contrast to the preponderance of the DQw3.2 allele in IDDM, the JRA patients expressed either the DQw3.1 or the DQw3.2 allele and sometimes both, without apparent association with disease expession. The different genomic markers reported here within HLA-DQ region potentially an analysis of HLA-associated function and disease susceptibility.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.23 no.5
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• pp.373-377
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• 1990

Volatile components of whole sardine sauce which was prepared with $7\%$ of complex enzyme-2000($2.18{\cdot}10^4\;U/g solid$), mixed with $6\%$ of invert sugar and heated at $90^{\circ}C$ for 2 hours were compared with those of without invert sugar. Thirty seven kinds were identified from the whole volatile components of hydrolysate heated without invert sugar and fourty three kinds were identified from the hydrolysate heated with $6\%$ of invert sugar. Amines were not detected from the whole volatile components of the chopped whole sardine hydrolysate. Considerable amount of 2,3-dihydrobenzofuran and 2-acetylpyrrole, a little amount of 2,5-hydrofuran, 2-ethylbutanol, 2-pyrone, 2-acetylfuran, 2,6-dimethylpyrazine, 2-acetylpyrazine, 5-methyl-2-furfural, furfuryl acetate, butylpyrrole and 2-methyl-3-hydroxypyrone were detected in the hydrolysate thermally treated with $6\%$ of invert sugar while these were not found in the hydrolysate heated without invert sugar. But the amount of 2-methyt-1-propa-not, hexane, butyl acetate and butyl alcohol were decreased, and acetic acid and butanoic acid were detected as volatile fatty acids.

• Journal of Periodontal and Implant Science
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• v.38 no.sup2
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• pp.343-354
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• 2008

Purpose: Aggregatibacter actinomycetemcomitans is associated with localized aggressive periodontitis. It produces cytolethal distending toxin (CDT), which induces cell cycle arrest in the G2/M phase. The CDT holotoxin is composed of CdtA, CdtB, and CdtC. CdtB has structural homology to human DNase I and is an active component of the CDT complex acting as a DNase. In particular, the pattern homology seen in the CdtB subunit has been associated with specific DNase I residues involved in enzyme catalysis, DNA binding, and metal ion binding. So, to study the functions and regulation of recombinant CdtB, we made up a quantity of functional recombinant CdtB and tested it in relation to the metal ion effect. Materials and Methods: We constructed the pET28a-cdtB plasmid from A. actinomycetemcomitans Y4 by genomic DNA PCR and expressed it in the BL21 (DE3) Escherichia coli system. We obtained the functional recombinant CdtB by the refolding system using the dialysis method and then analyzed the DNase activity and investigated the metal ion effect from plasmid digestion. Results: The recombinant CdtB subunit was expressed as the inclusion bodies. We were able to obtain functional recombinant CdtB subunit using refolding system. We confirmed that our refolded recombinant CdtB had DNase activity and was influenced by the metal ions $Mg^{2+}$ and $Ca^{2+}$. Conclusion: We suggest that the factors influencing recombinant CdtB may contribute to CDT associated diseases, such as periodontitis, endocarditic, meningitis, and osteomyelitis.

• Journal of Plant Biology
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• v.31 no.2
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• pp.91-100
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• 1988

The effect of Ca2+ and polyamines on the activity $\beta$-glucan synthetase II(GSII) related to cell wall synthesis was studied in carrot suspension cultured cells. The activity of GS II is four times higher than that of $\beta$-glucan synthetase I in carrot suspension cultured cells and in vitro expreiment, the activity of GSII was increased in response to increase in concentration of Ca2+ and polyamines. When carrot suspension cultured cells were incubated together with Ca2+ and polyamines, the GSII activity was high at 0.1mM of Ca2+ and 1mM of putrescine. Also, polycationic poly-L-lysine and poly-L-ornithine increased about 50% the GSII activity than that of the control, respectively. These results may imply that Ca2+ and polyamines were related to the enzyme activity as a polycationic nature. In addition, verapamil as the calcium channel blocker and flunarizine as an antagonist of calcium mechanism in cytoplasm decreased GSII activity ramarkably, Ca2+ and calmodulin stimulated GSII activity as Ca2+ of free ion rather than Ca2+ calmodulin complex. The effect of 2,4-D on the GSII activity in culture medium is shown to be low at 0.1mg per liter and GSII activity increased about 30% more than that of the 0.1mg/l at the range of 0.3-1.0mg per litere. Cummulative results suggest that Ca2+ and polyfamines stimulate the cell wall synthesis by means of the enhancement of GSII activity responsible for synthesizing the cell wall components.

• Molecules and Cells
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• v.19 no.3
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• pp.391-397
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• 2005

Molecular identification techniques are used where morphological characters are not useful for distinguishing species that resemble each other closely. The example studied here is the Adoxophyes species complex, in which A. orana (Fischer von $R{\ddot{o}}sslerstamm$) is officially the only known Korean species in the genus Adoxophyes (Lepidoptera: Tortricidae). However there have been suspicions that at least two types of A. orana exist in Korea based on the distribution and range of the host, with A. orana attacking apples and peaches, and another Adoxophyes sp. attacking tea and pears. The latter is presumed to be A. honmai (Yasuda), but the two have remained confused because of their extreme morphological similarity, despite several Asian studies of pheromonal and morphological characteristics. To confirm the occurrence of an Adoxophyes species other than A. orana in Korea, we compared 940 bp of the mitochondrial cytochrome oxidase I (COI) gene from 16 samples of Adoxophyes and found that there is a second Adoxophyes species different from A. orana. Comparison of the different sequences to that of Japanese A. honmai confirmed that they belong to the latter. From the sequence difference between the two Korean species, we were able to develop new PCR primer sets that distinguish them. This molecular identification technique with no enzyme digestion or sequencing step is a convenient and rapid way of differentiating between species that are hard to distinguish morphologically.

• The Korean Journal of Food And Nutrition
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• v.10 no.4
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• pp.528-532
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• 1997

A fermented squeezed-type paste was processed in order to highly effective utilization of cockle shell by-product, and improvement on rheological properties and texture of hydrolysate by used additives. The cockly shell by-products were homogenized with addition of water and enzymatically hydrolyzed at 5$0^{\circ}C$ for 8 hours added 4% Protease N.P.(Pacific Chemical Co.). And the hydrolysate was thermally treated for the purpose of flavor improvement, enzyme inactivation and pasteurization product at 10$0^{\circ}C$ for 1 hour, with 4% glucose. To make improvement of rheological properties, used complex additive with 0.5% alginic acid, 1% pectin and 0.2% agar were very effective. And stability of mixing was 98.1% after centrifuged at 10,000 rpm for 60 minutes. The chemical composition of moisture, total carbohydrate, total nitrogen and amino type nitrogen in the fermented squeeze-type cockle shell by-product paste were 57.7%, 20.6%, 1,458mg% and 1,187mg%, respectively. And the ratio of amino type nitrogen in total nitrogen was 81.4%.

• KSBB Journal
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• v.9 no.3
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• pp.312-318
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• 1994

The esterification of Ac-Tyr-OH was carried out in one-phase system containing ethanol by ${\alpha}$-chymotrypsin. The results of the esterification reaction are as follows. Chitin-${\alpha}$-chymotrypsin complex was found to be an effective catalyst for the esterlfication of Ac-Tyr-OH in ethanol organic solvent. The optimal conditions for the esterification were chitn/${\alpha}$-chymotrypsin ratio, 20(w/w); reaction temp., $35^{\circ}C$; reaction pH, 8.0; reaction time, 24 hrs. Also, addition of chitin in water/water-miscible organic solvent was effective for the stability of the enzyme. The esterification yield, Km and Vmax under optimal conditions were 93%, 3.093mM and 1.088mM/mg/hr, respectively.

• Korean Journal of Agricultural Science
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• v.21 no.1
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• pp.11-21
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• 1994

The halophile, Micrococcus sp. which produces RNase was isolated from salted and fermented food. The optimum growth condition of the Micrococcus sp. in pH 7.0 of complex medium containing 2M NaCl, and at $35^{\circ}C$. Optimum condition for enzyme production by this strain was when it was grown in the CM medium, containing 2% yeast extract, 1.5% casamino acid and 2M NaCl in the initial pH 8.5 for 2 days. The maximal RNase activity was observed at pH 8.0 and $55^{\circ}C$. The Km value for RNA was determined to be 5mg/ml by Lineweaver-Burk plot. The RNase activity in the absence of NaCl was maximum, but it was completely lost by adding of 1.25M NaCl and it was increased above 1.25M to 2.5M NaCl. When 2.5M NaCl was added, the activity of RNase showed 45% of maximum value.

• Biomedical Science Letters
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• v.15 no.1
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• pp.37-45
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• 2009

Insect cell cultures have become important tools in the production of biological substances for use in a variety of research, human and veterinary medicine, and pest control applications. These applications often require the introduction of foreign DNA into the cells and have generally used methods originally developed for use with human and other mammalian cell cultures. While these methods can be successfully employed, they are often less efficient with insect cells and frequently involve complex procedures or require specialized equipment. Even when they do work, they may require substantial modification because of differences in the culture medium or growth patterns of insect cells. In this study, We have optimized transfection conditions of Sf9 cell line using insect expression vector pIZT/V5-His which expresses green fluorescent protein effectively. Human stem cell factor (hSCF) is a glycoprotein that plays a key role in hematopoiesis acting both as a positive and negative regulator, often in synergy with other cytokines. It also plays a key role in mast cell development, gametogenesis, and melanogenesis. It can exist in membrane-bound form and in proteolytically released soluble form. As determined by an enzyme-linked immunosorbent assay performed, hSCF level in supernatant averaged 995ng/ml. The human hSCF was partially purified by immunoaffinity chromatography and analyzed with sodium dodecyl sulfate-polyacrylamide gel electrophoresis and immunoblotting. The results show that the hSCF has N-linked carbohydrate and corresponds to the soluble form, at or about 223 amino acids in length. The findings suggest functional importance for soluble hSCF in cells.