• Archives of Pharmacal Research
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• v.23 no.6
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• pp.605-612
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• 2000

Green tea polyphenols (GTP) have been demonstrated to suppress tumorigenesis in several chemical-induced animal carcinogenesis models, and predicted as promising chemopreventive agents in human. Recent studies of GTP extracts showed the involvement of mitogen-activated protein kinases (MAPKs) in the regulation of Phase II enzymes gene expression and induction of apoptosis. In the current work we compared the biological actions of five green tea catechins: (1) induction of ARE reporter gene, (2) activation of MAP kinases, (3) cytotoxicity in human hepatoma HepG2-C8 cells, and (4) caspase activation in human cervical squamous carcinoma HeLa cells. For the induction of phase IIgene assay, (-)-epigallocatechin-3-gallate (EGCG) and (-)-epicatechin-3-gallate (ECG) potently induced antioxidant response element (ARE)-mediated luciferase activity, with induction observed at 25 $\mu\textrm{m}$with EGCG. The induction of ARE reporter gene appears to be structurally related to the 3-gallate group. Comparing the activation of MAPK by the five polyphenols, only EGCG showed potent activation of all three MAPKs (ERK, JNK and p38) in a dose- and time-dependent manner, whereas EGC activated ERK and p38. In the concentration range of 25 $\mu\textrm{m}$ to 1 mM, EGCG and ECG strongly suppressed HepG2-ARE-C8 cell-growth. To elucidate the mechanisms of green tea polyphenol-induced apoptosis, we measured the activation of an important cell death protein, caspase-3 induced by EGCG, and found that caspase-3 was activated in a dose- and time-dependent manner. Interestingly, the activation of caspase-3 was a relatively late event (peaked at 16 h), whereas activation of MAPKs was much earlier (peaked at 2 h). It is possible, that at low concentrations of EGCG, activation of MAPK leads to ARE-mediated gene expression including phase II detoxifying enzymes. Whereas at higher concentrations of EGCG, sustained activation of MAPKs such as JNK leads to apoptosis. These mechanisms are currently under investigation in our laboratory. As the most abundant catechin in GTP extract, we found that EGCG potently induced ARE-mediated gene expression, activated MAP kinase pathway, stimulated caspase-3 activity, and induced apoptosis. These mechanisms together with others, may contribute to the overall chemopreventive function of EGCG itself as well as the GTP.

• Journal of the Korean Society of Food Science and Nutrition
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• v.40 no.11
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• pp.1501-1506
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• 2011

In this study, we optimized the reaction conditions for the bioconversion of green tea using tannase, and to evaluate its radical scavenging activities. Tea catechins such as (-)-epigallocatechin gallate (EGCG) or (-)-epicatechin gallate (ECG) were hydrolyzed by tannase to produce (-)-epigallocatechin (EGC) or (-)-epicatechin (EC), respectively, and a common product, gallic acid. The bioconversion of tea catechins by tannase was increased as enzyme concentration, substrate concentration and incubation time for enzyme dose. The results indicated the optimum reaction conditions for tannase were tannase 30 U/mL (enzyme concentration) on 1% green tea (substrate concentration) for 1 hr (incubation time for enzyme). Tannase enhanced the radical-scavenging properties of green tea; the 2,2-azinobis (3-ethylbenzthiazoline-6-sulfonic acid) (ABTS) and 1,1-diphenyl-2-picrylhydrazyl (DPPH) radicals scavenging abilities were significantly (p<0.001) greater for the tannase-treated green tea extract compared to the untreated green tea extract. It is reported that ECG has the greatest antioxidant activity among the catechins in green tea, and the release of gallic acid is considered to be beneficial because of its significant antioxidant potency. The results of this study suggest that the tannase-treated green tea increases antioxidant activities under optimum reaction conditions.

• Journal of Food Hygiene and Safety
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• v.22 no.3
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• pp.223-230
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• 2007

We have previously reported that green tea catechins(GTC) displayed potent antithrombotic effect, which was due to the antiplatelet activity. In the present study, the antiplatelet activity of each green tea catechin components was compared in vitro. Galloylated catechins including (-)-epigallocatechin gallate (EGCG), (-)-gallocatechin gallate (GCG), (-)-epicatechin gallate (ECG) and (-)-catechin gallate (CG), significantly inhibited collagen $(5{\mu}g/mL)-induced$ rabbit platelet aggregation with $IC_{50}$ values of 79.8, 63.0, 168.2 and $67.3{\mu}M$, respectively. EGCC GCG and CG also significantly inhibited arachidonic acid (AA, $100{\mu}M$)-induced rabbit platelet aggregation with $IC_{50}$ values of 98.9, 200.0 and $174.3{\mu}M$, respectively. However catechins without gallate moiety showed little inhibitory effects against rabbit platelet aggregation induced by collagen or AA compared with galloylated catechins. These observations suggest that the presence of gallate moiety at C-3 position may be essential to the antiplatelet activity of catechins and the presence of B ring galloyl structure may also contribute to the antiplatelet activity of GTC. In line with the inhibition of collagen-induced platelet aggregation, EGCG caused concentration-dependent decreases of cytosolic calcium mobilization, AA liberation and serotonin secretion. In contrast, epigallocatechin (EGC), a structural analogue of EGCG lacking a galloyl group in the 3' position, although slightly inhibited collagen-stimulated cytosolic calcium mobilization, failed to affect other signal transductions as EGCG in activated platelets. Taken together, these observations suggest that the antiplatelet activity of EGCG may be due to inhibition of arachidonic acid liberation and inhibition of $Ca^{2+}$ mobilization and that the antiplatelet of EGCG is enhanced by the presence of a gallate moiety esterified at carbon 3 on the C ring.

• Journal of the Korean Society of Food Science and Nutrition
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• v.37 no.8
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• pp.1025-1029
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• 2008

Fresh green tea leaf extracts were prepared by different enzyme treatment conditions, such as concentration, treating time and treating temperature using complex enzyme, Rapidase TF, and then extracted for 30 min at $80^{\circ}C$ to investigate their physicochemical properties. The results showed that free sugar content in every sample tended to increase, especially glucose content was increased up to 7.25 times compared to the control. Total amino acid was barely affected by the enzyme treatment and caffeine content was increased with reaction temperature. Total polyphenol and total catechin content was increased according to the amount of enzyme added and reaction temperature. Regardless of enzyme treatment conditions, composition of catechins were epigallocatechin, epicatechin, epicatechin gallate and epigallocatechin gallate by descending order of the content. Gallic acid content increased up to 0.04% and $45^{\circ}C$ with no further significant changes thereafter. From the results above, we could conclude that a simple and new method to extract green tea materials directly from fresh green tea leaves with improved extract ratio may be introduced by adding $0.08{\sim}0.1%$ of Rapidase TF to heat treated fresh green tea leaves and keeping temperature at $37{\sim}45^{\circ}C$ for $180{\sim}240\;min$ in order to skip existing complicated procedures.

• KOREAN JOURNAL OF CROP SCIENCE
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• v.53 no.2
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• pp.224-232
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• 2008

The health benefits associated with tea consumption have resulted in the wide inclusion of green tea extracts in botanical dietary supplements, which are widely consumed as adjuvants for complementary and alternative medicines. Tea contains polyphenols such as catechins or flavan-3-ols including (-)-epicatechin (EC), (-)-epigallocatechin (EGC), (-)-epicatechin gallate (ECG), and (-)-epigallocatechin gallate (EGCG), as well as the alkaloid, caffeine. The contents of catechins and caffeine in green tea are considered as a standard of quality evaluation of green tea. Therefor, the purpose of this study was to investigate the most suitable HPLC condition for simultaneous determination of catechins and caffeine in green tea extracts. The efficient HPLC analytical condition of catechins and caffeine contained green tea extracts was developed. The gradient elution employed a $250\;mm\;{\times}\;4.6\;mm$ i.d. YMC-pak ODS-AM 303 column. The gradient system was used two mobile phases. A gradient elution was performed with mobile phase A, consisting of 0.1% aqueous phosphoric acid, and mobile phase B, comprising 100% MeOH, and delivered at a flow rate of 1 mL/min as follows: $0{\sim}25\;min$, 80% A; $26{\sim}50\;min$, $80{\sim}70%$ A; 51 min, 80% A. $51{\sim}55\;min$, 80% A. The UV detection wavelength was set at 280 nm. The limit of detection (LOD) for catechins and caffeine standards were under 50 ng/mL.

• Journal of Life Science
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• v.21 no.9
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• pp.1219-1225
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• 2011

Phytochemicals, non-nutrient chemicals derived from plants, have been shown to have anti-inflammation, anti-oxidation, and chemopreventive effects. In the current study, we investigated whether five different phytochemicals (resveratrol, genistein, epicatechin gallate, diallyl disulfide, and caffeic acid phenethyl ester) alter cell growth and gene expression in human colorectal cancer HCT116 cells. Using a cell viability assay, we found that each of the phytochemicals tested inhibited HCT116 cell growth in a dose-dependent manner. Additionally, using human oligo DNA microarray analysis, we found that only six genes were commonly up-regulated and seven genes were commonly down-regulated in response to each phytochemical treatment. For the commonly up-regulated genes, the microarray analysis was confirmed by reverse transcription.PCR using gene-specific primers. In addition, NAG-1 protein was up-regulated by all treated phytochemcials. The results of this study may help to enhance our understanding of the general molecular mechanisms of chemoprevention that are mediated by phytochemicals in human colorectal cancer.

• Food Science and Preservation
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• v.16 no.5
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• pp.714-718
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• 2009

Polyphenols are naturally occurring antioxidants and play many important roles in human health. Grape leaves have not yet been reported to contain the antioxidants. In this study, the physicochemical properties of powders from young grape leaves, obtained after hot-air or vacuum freeze drying, were analyzed. Weight loss rangedfrom 76-80%, but no difference was found between samples prepared by hot-air orvacuum freeze drying. The L-, a-, and b- values of grape powder all fell upon hot-air drying. Quercetin, resveratrol, catechin, and epicatechin were detected in all cultivars. The contents of the four polyphenols were generally higher in powders obtained by vacuum freeze dryingthan in those prepared by hot-air drying, indicating that vacuum freeze drying is better for optimal preservation of polyphenols. None of three polyphenols assayed was found in some cultivars or after certain drying treatments. Based on physiochemical properties, our findings suggest that powders obtained after vacuum freeze drying are optimal for industrial use.

• Journal of Dairy Science and Biotechnology
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• v.27 no.2
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• pp.7-16
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• 2009

Appenzeller cheese samples were prepared by addition of 0.5, 1.0, and 2.0% green tea (Camellia sinensis, CS) powder and control cheese. We examined various quality characteristics of the novel cheese, such as viable-cell counts, pH, water-soluble nitrogen (WSN), non-casein nitrogen (NCN), non-protein nitrogen (NPN), and catechin level during maturation for 16 weeks at $14^{\circ}C$. To develop a Korean natural cheese containing green tea powder, we also analyzed the changes in the polyacrylamide gel electrophoresis pattern, chemical composition, and sensory qualities. The viable cell counts of the samples were not significantly different. Until the $3^{rd}$ week, the pH of the CS cheese decreased with an increase in the maturation time. However, the pH gradually increased by the $12^{th}$ week, while WSN, NCN, NPN also increased. The WSN, NCN, NPN, and catechin values for the CS cheese samples were significantly higher than the values for the control cheese. The polyacrylamide gel electrophoretic pattern of caseins for the CS cheese indicated that this cheese degraded more rapidly than the control cheese did. In the sensory evaluation, cheese with 1.0% CS powder showed the highest scores in taste and appearance and good scores in flavor and texture. These results indicate that 1.0% CS is the optimal value for addition to cheese, and cheese containing 1.0% CS shows good physiological properties and reasonably high overall sensory acceptability.