• Journal of Korean Biological Nursing Science
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• 제10권1호
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• pp.1-10
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• 2008

Purpose: The purpose of this study was to investigate the cerebral infarction size, antioxidant enzyme activities and lipid peroxidation changes after 6 weeks of dietary soybean protein intake in a rat focal brain ischemia model. Method: Weaning Sprague-Dawley rats were fed with either modified AIN-93G diet containing casein 20% (control), 20% soybean protein isolate-based diet (S20), or 40% of soybean protein isolate-based diet (S40) for 6 weeks. The animals were subject to right middle cerebral artery occlusion for 2 hr. After 24 hr of recirculation, the rats were sacrificed. Antioxidant enzymes activities of superoxide dismutase (SOD), catalase (CAT) and glutathione peroxidase (GPx) and thiobarbituric acid reactive substance (TBARS) level in the right brain were also measured. Result: There were no significant differences in the right cortical infarction volume, TBARS level, SOD and CAT activities among the three groups whereas the GPx activities of the S20 group were significantly higher than those of the control group (p=.02). Conclusion: Our results suggest that 20% of soybean protein may have a modulating effect on GPx and possibly have some protective effect against oxidative stress although it may enough to decrease cerebral infarction volume in rat focal brain ischemia model.

• International Journal of Oral Biology
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• 제30권3호
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• pp.77-84
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• 2005

EGCG[(-)-epigallocatechin gallate], is a major component of green tea has been considered as a major antioxidant constituent. It has been considered as potential chemopreventive and chemotherapeutic agents. However, very little is known about the cellular actions by which EGCG mediates its therapeutic effects. Various aspects of antioxidant activity of EGCG were evaluated in this study. EGCG itself did not show significant cytotoxicity. Significant 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical scavenging activity was observed in all ranges of concentration ($0.8-100{\mu}g/ml$) used in this study. Protective effect of EGCG against hydrogen peroxide induced cell death was observed. Relatively high lipid peroxidation inhibitory activity were detected ($IC_{50}$ was about $20{\mu}g/ml$). EGCG also dose-dependently enhanced the activities of superoxide dismutase (SOD), catalase (CAT) and glutathione peroxidase (GPX) in V79-4 cells. In concentrations of $100{\mu}g/ml$ of EGCG, activities of SOD, CAT and GPX were measured as 36.9 U/mg of protein, 22.9 U/mg of protein and 17.8 U/mg of protein, respectively. When these values were compared with those of the control groups (24.9 U/mg of protein, 14.9 U/mg of protein and 11.7 U/mg of protein), the relative increases were calculated as 48, 54 and 52%, respectively. Taken together, our findings suggest that EGCG can act as an antioxidant by scavenging radicals and enhancing antioxidant enzyme activities.

• Preventive Nutrition and Food Science
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• 제4권1호
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• pp.47-51
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• 1999

In order to evaluate the dietary regulation of cysteine sulfinic acid decarboxylase (EC 4.1.1.29) in cats, acitivity and protein content of CSAD were assessed in the liver and kidney of cats fed different levels of dietary protein, with and without taurine. Four groups of cats were fed one of the follow diets for 5 weeks ; 20% protein and taurine- free diet(LP0T) ; 20% protein and 0.15% taurine diet(LPNT) ; 60% protein and taurine-free diet(HP0T); and 60% protein and 0.15% taurine diet (HPNT). CSAD activity was determined in the liver and kidney of cats by measuring 14C2 released form [1-14C]-L cysteine sulfinic acid. CSAD protein was quantified using an immunochemical method. CSAD activity was extremely low in cat tissues, among which kidney showed the highest activity which was 0.118$\pm$0.050, and 0.377$\pm$0.056 nmol.min-1.mg soluble portein-1 iin animals fed LP0T and HP0T, respectively. Even though renal CSAD protein content was 18~55% of the hepatic CSAD protein content, renal CSAD acitivity was 1.3~6.5 times of the hepatic CSAD activity . Renal CSAD acitivities of cats fed 60% protein were about 1.6~3.2 times those of animals fed 2.% protein , and hepatic CSAD activity was not significantly affected by the dietary level of protein. Taurine depletion significantly elevated both hepatic and renal CSAD activities above the values for cats having normal taurine status most probably as an adaptive response.

• KSBB Journal
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• 제23권3호
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• pp.231-238
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• 2008

암세포의 침윤은 숙주 조직의의 기저막과 세포외 기질을 침투함으로써 일어난다. 침윤과 전이과정에는 단백질가수분해 효소인 matrix metalloproteinases (MMPs)가 깊이 연관되어 있는 것으로 알려져 있으며, MMP의 가수분해 활성은 tissue inhibitors of metalloproteinases (TIMPs)라는 억제 단백질에 의해 억제된다. TIMP-2는 21kDa 크기의 포유류 단백질로 대장균에서 과발현 시 다른 많은 포유류 단백질과 마찬가지로 가용성이 낮은 봉입체 형태로 발현된다. TIMP-2 단백질의 접힘에 6개의 이황화결합이 필요하고, 이는 일반적으로 대장균 환경은 적합하지 않다. 본 연구에서는 대장균에서 불가용성으로 발현되는 TIMP-2 유전자를 유전자셔플링 기법의 한 가지인 StEP (staggered extension process)를 변형하고 동시에 $Mn^{2+}$ 농도 변화와 dGTP 불균형을 이용한 무작위 돌연변이 기법을 혼용하여 대장균에서 가용성 TMP-2 재조합 변이체를 생성하고자 하였다. 무작위로 재조합된 TIMP-2 유전자 중에서 가용성으로 발현되는 TIMP-2 유전자를 선별하기 위해서 chloramphenicol acetyltransferase (CAT)-융합 방법을 도입하였다. CAT 유전자가 가용성으로 발현되는 재조합 TIMP-2 유전자에 융합되면 이를 갖는 E. coli는 높은 chloramphenicol 환경에서 생존이 가능하게 된다. 이러한 in virro mutageuesis 기법과 CAT-융합 방법으로 대장균 가용성 TIMP-2 재조합 변이체를 14가지 얻을 수 있었다. 변이체 TIMP-2의 아미노산서열 분석과 구조 분석 결과 주로 소수성 아미노산이 친수성 아미노산으로 전환되었고, MMP와의 결합이 관여하지 않는 C-말단 부위에 돌연변이가 집중되어 있었다. 본 연구에서 개발된 간편하고 새로운 in vitro 재조합 방법과 CAT을 이용한 스크리닝 기법은 다른 많은 대장균 내 불가용성 단백질의 발현에도 사용될 수 있을 것으로 사료된다.

• Natural Product Sciences
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• 제9권3호
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• pp.186-191
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• 2003

The hepatoprotective effect of methanol extract of leaves of Caesalpinia bonducella was studied by means of paracetamol induced liver damage in rats. The degree of protection was measured by using biochemical parameters such as serum transaminase (SGPT and SGOT), alkaline phosphatase (ALP), bilirubin, and total protein. Further, the effects of the extract on lipid peroxidation (LPO), glutathione (GSH), superoxide dismutase (SOD), and catalase (CAT) were estimated. The methanol extract of C. bonducella (MECB) (50,100 and 200 mg/kg) produced significant (P<0.01) hepatoprotective effect by decreasing the activity of serum enzymes, bilirubin, and lipid peroxidation, while it significantly increased increased the levels of GSH, SOD, CAT, and protein in a dose dependent manner. The effects of MECB were comparable to that of standard drug Silymarin. However, at a lower dose (25 mg/kg) it could not restore the deleterious effect produced by paracetamol. The results indicate that Caesalpinia bonducella had antioxidant and hepatoprotective effects.

• BMB Reports
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• 제44권10호
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• pp.647-652
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• 2011

The protein transduction domains have been reported to have potential to deliver the exogenous molecules, including proteins, to living cells. However, poor transduction of proteins limits therapeutic application. In this study, we examined whether imipramine could stimulate the transduction efficiency of PEP-1 fused proteins into astrocytes. PEP-1-catalase (PEP-1-CAT) was transduced into astrocytes in a time- and dose-dependent manner, reducing cellular toxicity induced by $H_2O_2$. Additionally, the group of PEP-1-CAT + imipramine showed enhancement of transduction efficiency and therefore increased cellular viability than that of PEP-1-CAT alone. In the gerbil ischemia models, PEP-1-CAT displayed significant neuroprotection in the CA1 region of the hippocampus. Interestingly, PEP-1-CAT + imipramine prevented neuronal cell death and lipid peroxidation more markedly than PEP-1-CAT alone. Therefore, our results suggest that imipramine can be used as a drug to enhance the transduction of PEP-1 fusion proteins to cells or animals and their efficacies against various disorders.

• 대한약학회:학술대회논문집
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• 대한약학회 2003년도 Proceedings of the Convention of the Pharmaceutical Society of Korea Vol.2-2
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• pp.77.3-78
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• 2003

We previously shown that sphingosylphosphorylcholine, a lysophosphatidic acid, produced contraction in isolated single cells of cat ilium. We investigated the mechanism of sphingosine-1-phosphate (S1P)-induced contraction of circular smooth muscle cells in cat esophagus. S1P produced esophageal contraction in a dose dependent manner. The maximal contraction (l0$\^$-7/ M) induced at 1min. Pertusis toxin (PTX) inhibited contraction induced by S1P, suggesting that the contraction is mediated to a PTX-sensitive G-protein. (omitted)

• 생약학회지
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• 제50권4호
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• pp.285-290
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• 2019

This study was carried out to identify the skin anti-aging effect of cycloheterophyllin on dermal fibroblasts. To elucidate anti-aging effects of cycloheterophyllin on dermal fibroblasts, I measured cell viability, mRNA expressions, and Collagen, type I/matrix metallopeptidase 1(MMP1)-ELISA assay. In this study, I investigated the effects of cycloheterophyllin on Collagen, type I, alpha 1(COL1A1)/Collagen, type III, alpha 1(COL3A1)/MMP1/Superoxide dismutases/Catalase(CAT) mRNA expressions and Collagen, type I/MMP1 protein production. Quantitative Real-time RT-PCR showed that cycloheterophyllin increased mRNA level of COL1A1/COL3A1/CAT genes and collagen, type I protein by ELISA assay compared to UVA-treated dermal fibroblasts. Furthermore MMP1 mRNA and protein expressions were decreased by cycloheterophyllin treatment. These observations revealed that cycloheterophyllin increased anti-aging effects in dermal fibroblasts. Therefore, I identified the anti-aging effects of cycloheterophyllin, and these results showed that the cycloheterophyllin can be a considerable potent ingredient for skin anti-aging. Based on this, I anticipated further researches about cycloheterophyllin for mechanism to develop not only cosmetics but for healthcare food or medicine.

• 생명과학회지
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• 제6권4호
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• pp.241-249
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• 1996

Vivrio mimicus (ATCC 33568)의 최적 배양조건하에서 배양액으로부터 collagenase를 황산암모늄 염석과 DEAE-Sephadex A-50 이온코환크로마토그래피에 의해 분리. 정제하였다. SDS-PAGE 전기영동분석법 및 겔여과법으로 정제된 collagenase의 분자량은 42 kDa 이였다. 기질인 불용성 콜라겐(Type I)에 대한 collagenase의 최적 pH 및 온도는 각각 7.75 및 28$\circ$였다. 금속착물제와 serine protease 저해제는 collagenase의 활성을 저해하였지만 L-cysteine과 histidine은 효소의 활성을 저해하지 않았다. collagenase의 아미노산 조성은 glycine 및 alanine의 아미노산 잔기가 많이 함유되어 있었다. 불용성 (Type I) 콜라겐에 대한 collagenase의 속도상수인 K$_{m}$ 값 및 R$_{cat}$/K$_{m}$값은 각각 2.86 mg/ml 및 972.28 U/mg-protein 이었다.

• International Journal of Industrial Entomology and Biomaterials
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• 제4권1호
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• pp.63-68
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• 2002

A cDNA encoding a putative member of cathepsin B of the thiol pretense superfamily was cloned from a cDNA library of the mulberry longicorn beetle, Apriona germari. Sequence analysis of the cDNA encoding the cathepsin B of A. germari (AgCatB) revealed that the 972 bp cDNA has an open reading frame of 324 amino acid residues. The deduced protein sequence of the AgCatB showed high homology with cathepsin B of the insects, Bombyx mori (47.3% amino acid identity), Helicoverpa armigera (46.6%) and Sarcophaga peregrina (45.6%), and the lowest homology with Aedes aegypti (33.2%). The AgCatB contains six disulfate bonds typical for cysteine pretenses. The three amino acid positions Cys-109, His-267, and Asn-287 which are conserved, active sites characteristic for cathepsin B, were also found. Phylogenetic analysis further confirmed that the AgCatB has a close relationship with that of B. mori, H. armigera and S. peregrina.