• Proceedings of the Korean Society of Applied Pharmacology
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• 1993.04a
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• pp.134-134
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• 1993

Tn3 Transposon을 이용한 Shuttle Mutagenesis방법으로 효모의 Genome 상에 무작위적으로 $\beta$-gal 표지 유전자를 삽입하고 효모생활사의 각 세포시기 마다 특이하게 발현되는 유전자를 X-Gal plate상에서 찾아내고 이들 효모 유전자의 단백질이 세포내에 어떤 위치에 분포하는가를 간접 면역현미경법으로 추적해보았다. 먼저 효모의 genomic library를 38bp의 Tn3 Termial repeat를 가지고 있지 않은 pHSS6 Vector에 patial fill-in 방법으로 조성하였으며 최종적으로 20 Genome equivalent에 해당하는 18개 Pool의 genomic library를 만들었다. 이들 library를 조사하여 본 결과 모든 클론이 평균 3kb 크기의 insert를 가지고 있었으며 이는 99.99%의 효묘 genome을 대표하였다. 특정한 유전자의 발현을 알아보기 위해 먼저 mini-Tn3로 shuttle mutagenesis를 실시하고 vegetative growth동안 발현되는 유전자를 X-gal을 사용하여 골라내었다. 지금까지 16823개의 클론을 조사하였는데 이중 13%에 해당하는 2187개가 X-gal plate상에서 양성반응을 보여주었다. 양성반응을 보여주는 융합단백질의 세포내 분포틀 anti-$\beta$-galactosidase 항체를 사용하여 추적해보았다. 항체론 이용한 형광염색결과 약 70%의 세포가 background이상의 염색을 보여주었으며 이중 novel한 염색 pattern을 나타내는 클론도 다수 탐지되었다. 이상의 결과를 종합하면 Tn3틀 이용한 Shuttle Mutagenesis 방법으로 지금까지 전통적인 유전학적인 접근 방식으로 탐지되지 않았던 다수의 새로운 효모 유전자를 찾아낼 수 있을 것으로 사료된다.tamine제중 triprolidine이 $K_{M}$ /K$_{H}$ 비가 가장 높았고 diphenidol이 가장 낮았다. 이상의 결과로 보아 항 histamine제의 muscarinic receptor 차단작용은 이들 약물의 항 alleragy 효과에 필요한 작용이 아니며 본 실험에서 추정된 항 histamine제의 H$_1$-receptor와 muscarinic receptor에 대한 상대적 역가는 이들 약물의 선택과 평가에 중요한 지표가 될수 있을 것으로 생각된다.ing ischemic insults. The nature of the receptor is being explored by molecular genetic techniques, and we have recently cloned two of the major subunits; some of the data will be presented.LIFO, 우선 순위 방식등을 선택할 수 있도록 확장하였다. SIMPLE는 자료구조 및 프로그램이 공개되어 있으므로 프로그래머가 원하는 기능을 쉽게 추가할 수 있는 장점도 있다. 아울러 SMPLE에서 새로이 추가된 자료구조와 함수 및 설비제어 방식등을 활용하여 실제 중형급 시스템에 대한 시뮬레이션 구현과 시스템 분석의 예를 보인다._3$", chain segment, with the activation energy of carriers from the shallow trap with 0.4[eV], in he amorphous regions.의 증발산율은 우기의 기상자료를 이용하여 구한 결과 0.05 - 0.10 mm/hr 의 범위로서 이로 인한 강우손실량은 큰 의미가 없음을 알았다.재발이 나타난 3례의 환자를 제외한 9례 (75%)에서는 현재까지 재발소견을 보이지 않고 있다.

• Molecular & Cellular Toxicology
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• v.3 no.2
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• pp.107-112
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• 2007

o-Nitrotoluene is used to synthesize artificial dyes and raw materials of urethane resin. In this study, we have carried out in vitro genetic toxicity tests and microarray analysis to understand the underlying mechanisms and the mode of action of toxicity of onitrotoluene. TA1535 and TA98 cells were treated with o-nitrotoluene to test its toxicity by basic genetic toxicity test. Ames and two new in vitro micronucleus and COMET assays were applied using CHO cells and L5178Y cells, respectively. In addition, microarray analysis of differentially expressed genes in L5178Y cells in response to o-nitrotoluene was analyzed using Affymatrix genechip. The result of Ames test was that o-nitrotoluene treatment did not increase the mutations both in base substitution strain TA1535 and in frame shift TA98. o-Nitrotoluene has not increased micronuclei in CHO cells. But onitrotoluene increased DNA damage in L5178Y cell. Two-hundred two genes were initially selected as differentially expressed genes in response to o-nitrotoluene by microarray analysis and forty four genes among them were over 2 times of log fold changed. These forty four genes could be candidate biomarkers of genetic toxic action of o-nitrotoluene related to induction of mutation and/or induction of micronuclei and DNA damage. Further confirmation of these candidate markers related to the DNA damage will be useful to understand the detailed mechanism of action of o-nitrotoluene.

• Molecular & Cellular Toxicology
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• v.3 no.2
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• pp.90-97
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• 2007

8-Hydroxyquinoline is used as antibacterial agent and antioxidant based on its function inducing the chelation of ferrous ion present in host resulting in production of chelated complex. This complex being transported to cell membrane of bacteria and fungi exerts antibacterial and antifungal action. In this study, we have carried out in vitro genetic toxicity tests and microarray analysis to understand the underlying mechanisms and the mode of action of toxicity of 8-hydroxyquinoline. TA1535 and TA98 cells were treated with 8-hydroxyquinoline to test its toxicity by basic genetic toxicity test, Ames and two new in vitro micronucleus and COMET assays were applied using CHO cells and L5178Y cells, respectively. In addition, microarray analysis of differentially expressed genes in L5178Y cells in response to 8-hydroxyquinoline were analyzed using Affymatrix genechip. The result of Ames test was that 8-hydroxyquinoline treatment increased the mutations in base substitution strain TA1535 and likewise, 8-hydroxyquinoline also increased mutations in frame shift TA98. 8-Hydroxyquinoline increased micronuclei in CHO cells and DNA damage in L5178Y. 8-Hdroxyquinoline resulted in positive response in all three tests showing its ability to induce not only mutation but also DNA damage. 783 Genes were initially selected as differentially expressed genes in response to 8-hydroxyquinoline by microarray analysis and 34 genes among them were over 4 times of log fold changed. These 34 genes could be candidate biomarkers of genetic toxic action of 8-hydroxyquinoline related to induction of mutation and/or induction of micronuclei and DNA damage. Further confirmation of these candidate markers related to their biological function will be useful to understand the detailed mode of action of 8-hydroxyquinoline.

• The Korean Journal of Mycology
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• v.20 no.4
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• pp.324-336
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• 1992

On the five interspecific protoplast fusants of Ganoderma lucidum and G. applanatum was the antitumor test performed. The fusant F-2 was selected, to examine the cultured mycelia (protein bound polysaccharide) as antitumor components. When a dose of 20 mg/kg/day of each components purifed from F-2 fusant was, i.p., injected into ICR mice, the inhibition ratio of Fr. II against the solid form of sarcoma 180 increased to 1.5 times as compared with that of their parents. When Fr. II was examined for immunopotentiation activity, it increased the amount of the superoxide anion in activated macrophages to 1.2 times and the count of hemolytic plaque forming cells in the spleen to 4.3 times as compared with that of each control group. Its chemical analysis showed 85.2% polysaccharide which consisted of glucose, galactose, mannose, fucose and xylose, and 0.39% protein of 15 amino acids. The content of hexosamine was 0.39% and the molecular weight of Fr. V was $5.6{\times}10^4$ dalton.

• Journal of the Korea Academia-Industrial cooperation Society
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• v.15 no.8
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• pp.5088-5094
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• 2014

Hovenia dulcis extract (HDE) has positive effects on alcohol degradation, recovery of liver damage and antioxidant activities. This study examined whether HDE exerts an ameliorative effect on inflammatory orifacial pain in an animal algesic model with formalin. The animals (rats) were divided into four groups: group I (control), group II (right facial subcutaneous injection of 5% formalin, inflammatory orifacial pain group), group III (5% formalin + distilled water administration), and group IV injection (5% formalin + 4.5 ml/kg of HDE), respectively. The scores from the scratch and effleurage tests were applied to evaluate the differences between three groups. The expression of p38 MAPK, iNOS and Nrf2 in the brain and medulla oblongata, which are involved in pain regulation, inflammation, antioxidation and nitric oxide production, were analyzed by western blot. The degree of orifacial pain was significantly lower in group IV than in groups I, II and, III. The expression of p38MAPK, iNOS and Nrf2 in the brain and medulla were also lower in group IV than in the other groups. These findings suggested that a Hovenia dulcis extract can attenuate inflammatory orifacial pain by suppressing the expression of p38 MAPK, iNOS and Nrf2.

• Food Science of Animal Resources
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• v.23 no.4
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• pp.299-308
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• 2003

Quality characteristics of sausage added with green tea powder was studied to investigate the substitutive effect of green tea for nitrite. Residual nitrite, thiobarbituric acid reactive substances(TBARS) and volatile basic nitrogen(VBN) contents of 9 different sausages prepared with 3 levels of nitrite(50, 100 or 150 ppm) and 3 levels of green tea(0, 0.5 or 1％) were measured during storage at l0$^{\circ}C$ for 6 weeks or 20$^{\circ}C$ for 14 days. Proximate compositions of the sausage were not affected by the addition level of green tea or nitrite. The lightness(L value) and redness(a value) of sausage decreased, while yellowness(b value) increased, with the addition of green tea powder. Residual nitrite content was lower in sausages containing green tea when prepared with same amount of nitrite, showing the addition of green tea powder has a beneficial effect on reducing the residual nitrite content in sausage. TBARS and VBN contents were lower in sausages with nitrite and green tea than those with nitrite alone. The TBARS and VBN lowering effects of green tea were higher in sausages prepared with low level of nitrite than those prepared with high amount of nitrite. The results suggest that the addition of green tea powder may reduce the residual nitrite content of sausage, and the use of green tea may also cut down the use of nitrite in the production of pork sausage with little deterioration in preservation.

• Korean Journal of Food Science and Technology
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• v.33 no.3
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• pp.338-344
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• 2001

The effect of gamma-irradiation on the microbiological and general qualities of Kanjang (fermented soy sauce, Korean type) and Shoyu (fermented soy sauce, Japanese type) was studied. Samples were prepared, irradiated at 5, 10 and 20 kGy, and then stored at $25^{\circ}C$ for 18 weeks. The results showed that the Bacillus spp. was inactivated by 3 log cycles with the irradiation dose of 10 kGy and the number of Bacillus cells that survived from 10 to 20 kGy of gamma irradiation was decreased gradually during storage. Yeasts and Lactobacillus were nearly eliminated at 5 to 10 kGy of irradiation. The $D_{10}$ values of Bacillus, yeast and Lactobacillus in Shoyu were 2.67 kGy, 0.81 kGy and 1.30 kGy, in Kanjang were 2.75 kGy, 0.99 kGy and 1.47 kGy, respectively. The general quality of gamma irradiated Shoyu and Kanjang, such as total nitrogen, amino nitrogen, protease activity and pH were more stable than that of control during storage periods. Decolorization was observed just after irradiation, but the color was recovered during storage. The sensory evaluations showed that irradiated samples were more acceptable. Therefore, it was considered that gamma irradiation was effective for maintaining better quality of Shoyu and Kanjang during storage.

• Journal of Aquaculture
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• v.21 no.3
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• pp.176-180
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• 2008

This study clarified biochemical overripeness characterization of ovulated eggs of Anguilla japonica and suggested a method maintained overripeness after ovulation for high hatching rates. In maturated Japanese eel eggs, the relationships between fertilization rate and hatching rate, and fertilization and survival rates were measured. DNA contents showed the significantly low 0.653 pg/ug protein in 20% downward hatching rate trial with decrease of hatching rate(P<0.05), whereas RNA/DNA ratio showed the significantly high 1.058 in 20% downward hatching rate trial(P<0.05). And activities of total alkaline protease and ACPase according to the hatching rate groups did not show the significant difference(P>0.05). The protein contents were assayed the significantly high 186.16 ug/mg protein in 20% downward hatching rate trial(P<0.05). However, the overripened eggs had lowed hatching rate, because of stimulate the overripening of normal maturated eggs due to the continuous supplement of protein (vitellogenin). We suggested that need to reduce supplement speed or interception of vitellogenin produced in live for prevent overripeness of maturated eggs after ovulation

• Tuberculosis and Respiratory Diseases
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• v.53 no.2
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• pp.101-112
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• 2002

Background : Toluene diisocyanate(TDI) is a leading cause of occupational asthma. However, the pathogenesis of TDI-induced asthma is largely unknown because there is no suitable animal model. Materials and Methods : We developed a murine model of TDI-induced asthma by performing two sensitization with 3% TDI and one challenge with 1% TDI using ultrasonic nebulization. Results : Similar to occupational asthma in humans, murine TDI-induced asthma includes findings 1) increased inflammatory cells, including neutrophils and eosinophils, 2) histologic changes, including infiltration of inflammatory cells around bronchioles, thickened airway epithelium, contraction of bronchioles, and accumulation of mucus and debris in the bronchioles, 3) increased MMP-9 activity in inflammatory cells in the airway lumen, 4) airway hyperrespnosiveness. Administraion of an MMP inhibitor, MMPI-I, remarkably reduced all these pathophysiological findings. Conclusion : Therefore, we conclude that TDI-induced occupational asthma is associated with the induction of MMP-9 in inflammatory cells, and the inhibition of MMP-9 may be a good therapeutic strategy.

• The Korean Journal of Mycology
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• v.37 no.1
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• pp.91-95
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• 2009

To examine physico-chemical properties of the polysaccharide extracted from liquid-cultured mycelia of Schizophyllum commune, each the polysaccharide was extracted with hot water treatment and then fractionated with ethanol, alkaline solution and ultrafiltration. And we determined carbohydrate contents, composition of amino acids, infra-red spectrum and viscosity. Carbohydrate contents of polysaccharide treated with ethanol and ultrafiltration were 72.0% and 62.3%, and proteins content were 15.3% and 32.0% respectively. The carbohydrate consisted of four monosaccharides and the protein contained 16 amino acids. The polysaccharide obtained from ultrafiltration was shown an absorption band characteristic of the ${\beta}$-glycosidic linkage by infra red spectra. These results suggest that the polysaccharide extracted from Schizophyllum commune showed the characteristics of proteinbounded polysaccharide, and it was ${\beta}$-glycosidic linkage with strong viscosity.