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Activation of Antioxidant-Response Element (ARE), Mitogen- Activated Protein Kinases (MAPKs) and Caspases by Major Green Tea Polyphenol Components during Cell Survival and Death

  • Chen, Chi;Yu, Rong;Owuor, Edward D.;Kong, A.NTony
    • 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.

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Physicochemical and Microbiological Properties of Ginseng-Whey Beverages (인삼 유청음료의 이화학적 및 미생물학적 특성)

  • 기해진;홍윤호
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.22 no.2
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    • pp.208-214
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    • 1993
  • Ginseng-whey beverages were prepared with rennet whey, ginseng, sweetener, honey and Japanese apricot, inoculated with different strains of lactic acid bacteria or unfermented partly. The samples were stored at 4$^{\circ}C$ or 30$\pm$1$0^{\circ}C$ and then physicochemical and microbiological properties were investigated. The yield of whey was 78.8%. The pH-values reduced and acidities increased during the storage period. The contents of solid-substances, ash and lipid in ginseng-whey beverages were 7.90~8.20%, 0.62~0.66% and 0.16%, respectively. The protein contents of ginseng-whey beverages were 0.42~0.56% and the contents were not changed during the storage period. The lactose contents of fermented beverages were higher than those of unfermented beverages. During the storage period (1~5 weeks), the ranges of D(-) - and L(+)- lactic acid contents in fermented ginseng-whey beverages (17.3~156.1 mg/100g, 347.3~1894.2mg/100g) were higher than those of unfermented ginseng-whey beverages (6.2~82.8mg/100g, 7.1~885.5mg/100g). The contents of total saponin in unfermented sample and fermented sample (Lac. casei sub-sp. casei+Str. salivarius sub-sp. thermophilus) were increased during the storage period. But, those of the fermented sample(Lac. acidophilus+Lac. delbrueckii sub-sp. bulgaricus) were reduced. In the electrophoretic results of ginseng-whey beverages, an $\alpha$-lactalbumin and a $\beta$-lactoglobulin bands were shown apparently and there were no changes observed during the storage period. During the storage period (1~3 week) the coliform was not detected and total plate counts and psychrotrophs were increased according to the storage period.

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Relationship between Rainfall and Zooplankton Community Dynamics in a Riverine Wetland Ecosystem (Upo) (강 배후 습지생태계(우포)에서 강우량과 동물플랑크톤 군집 동태)

  • Kim, Hyun-Woo;Choi, Jong-Yun;La, Geung-Hwan;Jeong, Kwang-Seuk;Jo, Gea-Jae
    • Korean Journal of Ecology and Environment
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    • v.43 no.1
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    • pp.129-135
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    • 2010
  • The relationship between rainfall variable and zooplankton dynamics was studied in the Upo wetland, an ecosystem of international importance. Water sampling was conducted on biweekly basis from January 2002 to December 2007 in the study site. The annual average of total rainfall was 1,324 mm during the study period. Total rainfall amount in 2003 (1,766 mm) was unusually high, while total rainfall amount in 2005 (975 mm) was exceptionally lower than the average. Most of basic limnological parameters (water temperature, dissolved oxygen, pH, conductivity and turbidity) in the study site were greatly influenced by the flooding events and rainfall amounts in summer. There were statistically significance between seasonal and inter-annual differences in zooplankton abundance and the total rainfall amount (ANOVA, P<0.05). Zooplankton abundance was high in summer (mean${\pm}$s.d.: $1,594{\pm}1,598\;Ind.\;L^{-1}$) and low in winter ($246{\pm}234\;Ind.\;L^{-1}$. The 47% of annual total zooplankton abundance in the study site were observed in summer. The seasonal pattern of rotifers was similar to that of total zooplankton. This reflected the fact that rotifers strongly dominated and occupied ca. 65% the total zooplankton abundance (annual mean: $398{\pm}1,139\;Ind.\;L^{-1}$, n=149), followed by cladocerans ($65{\pm}140\;Ind.\;L^{-1}$) and copepods ($58{\pm}84\;L^{-1}$). Planktonic rotifers such as Keratella cochlearis, Polyarthra spp. and Brachionus calyciflorus were dominant from winter to spring and attached rotifers such as Lecane spp., Monostyla spp. and Trichocerca spp., observed commonly from spring to fall. Among the environmental variables considered, rainfall in summer seemed to play the most important role in determining characteristics of zooplankton community dynamics in the Upo wetland.

An Analytical Validation of the GenesWellTM BCT Multigene Prognostic Test in Patients with Early Breast Cancer (조기 유방암 환자를 위한 다지표 예후 예측 검사 GenesWellTM BCT의 분석적 성능 시험)

  • Kim, Jee-Eun;Kang, Byeong-il;Bae, Seung-Min;Han, Saebom;Jun, Areum;Han, Jinil;Cho, Min-ah;Choi, Yoon-La;Lee, Jong-Heun;Moon, Young-Ho
    • Korean Journal of Clinical Laboratory Science
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    • v.49 no.2
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    • pp.79-87
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    • 2017
  • GenesWell$^{TM}$ BCT is a 12-gene test suggesting the prognostic risk score (BCT Score) for distant metastasis within the first 10 years in early breast cancer patients with hormone receptor-positive, HER2-negative, and pN0~1 tumors. In this study, we validated the analytical performance of GenesWell$^{TM}$ BCT. Gene expression values were measured by a one-step, real-time qPCR, using RNA extracted from FFPE specimens of early breast cancer patients. Limit of Blank, Limit of Detection, and dynamic range for each of the 12 genes were assessed by serially diluted RNA pools. The analytical precision and specificity were evaluated by three different RNA samples representing low risk group, high risk group, and near-cutoff group in accordance with their BCT Scores. GenesWell$^{TM}$ BCT could detect gene expression of each of the 12 genes from less than $1ng/{\mu}L$ of RNA. Repeatability and reproducibility across multiple testing sites resulted in 100% and 98.3% consistencies of risk classification, respectively. Moreover, it was confirmed that the potential interference substances does not affect the risk classification of the test. The findings demonstrate that GenesWell$^{TM}$ BCT have high analytical performance with over 95% consistency for risk classification.

Omega-7 producing alkaliphilic diatom Fistulifera sp. (Bacillariophyceae) from Lake Okeechobee, Florida

  • Berthold, David Erwin;Rosa, Nina de la;Engene, Niclas;Jayachandran, Krish;Gantar, Miroslav;Laughinghouse, Haywood Dail IV;Shetty, Kateel G.
    • ALGAE
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    • v.35 no.1
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    • pp.91-106
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    • 2020
  • Incorporating renewable fuel into practice, especially from algae, is a promising approach in reducing fossil fuel dependency. Algae are an exceptional feedstock since they produce abundant biomass and oils in short timeframes. Algae also produce high-valued lipid products suitable for human nutrition and supplement. Achieving goals of producing algae fuels and high-valued lipids at competitive prices involves further improvement of technology, especially better control over cultivation. Manipulating microalgae cultivation conditions to prevent contamination is essential in addition to promoting optimal growth and lipid yields. Contamination of algal cultures is a major impediment to algae cultivation that can however be mitigated by choosing extremophile microalgae. This work describes the isolation of alkali-tolerant / alkaliphilic microalgae native to South Florida with ideal characteristics for cultivation. For that purpose, water samples from Lake Okeechobee were inoculated into Zarrouk's medium (pH 9-12) and incubated for 35 days. Selection resulted in isolation of three strains that were screened for biomass and lipid accumulation. Two alkali-tolerant algae Chloroidium sp. 154-1 and Chlorella sp. 154-2 were poor lipid accumulators. One of the isolates, the diatom Fistulifera sp. 154-3, was identified as a lipid accumulating, alkaliphilic organism capable of producing 0.233 g L-1 d-1 dry biomass and a lipid content of 20-30% dry weight. Lipid analysis indicated the most abundant fatty acid within Fistulifera sp. was palmitoleic acid (52%), or omega-7, followed by palmitic acid (17%), and then eicosapentanoic acid (15%). 18S rRNA phylogenetic analysis formed a well-supported clade with Fistulifera species.

Selection for Duration of Fertility and Mule Duck White Plumage Colour in a Synthetic Strain of Ducks (Anas platyrhynchos)

  • Liu, H.C.;Huang, J.F.;Lee, S.R.;Liu, H.L.;Hsieh, C.H.;Huang, C.W.;Huang, M.C.;Tai, C.;Poivey, J.P.;Rouvier, R.;Cheng, Y.S.
    • Asian-Australasian Journal of Animal Sciences
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    • v.28 no.5
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    • pp.605-611
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    • 2015
  • A synthetic strain of ducks (Anas platyrhynchos) was developed by introducing genes for long duration of fertility to be used as mother of mule ducklings and a seven-generation selection experiment was conducted to increase the number of fertile eggs after a single artificial insemination (AI) with pooled Muscovy semen. Reciprocal crossbreeding between Brown Tsaiya LRI-2 (with long duration of fertility) and Pekin L-201 (with white plumage mule ducklings) ducks produced the G0. Then G1 were intercrossed to produce G2 and so on for the following generations. Each female duck was inseminated 3 times, at 26, 29, and 32 weeks of age. The eggs were collected for 14 days from day 2 after AI. Individual data regarding the number of incubated eggs (Ie), the number of fertile eggs at candling at day 7 of incubation (F), the total number of dead embryos (M), the maximum duration of fertility (Dm) and the number of hatched mule ducklings (H) with plumage colour were recorded. The selection criterion was the breeding values of the best linear unbiased prediction animal model for F. The results show high percentage of exhibited heterosis in G2 for traits to improve (19.1% for F and 12.9% for H); F with a value of 5.92 (vs 3.74 in the Pekin L-201) was improved in the G2. Heritabilities were found to be low for Ie ($h^2=0.07{\pm}0.03$) and M ($h^2=0.07{\pm}0.01$), moderately low for Dm ($h^2=0.13{\pm}0.02$), of medium values for H ($h^2=0.20{\pm}0.03$) and F ($h^2=0.23{\pm}0.03$). High and favourable genetic correlations existed between F and Dm ($r_g=0.93$), between F and H ($r_g=0.97$) and between Dm and H ($r_g=0.90$). The selection experiment showed a positive trend for phenotypic values of F (6.38 fertile eggs in G10 of synthetic strain vs 5.59 eggs in G4, and 3.74 eggs in Pekin L-201), with correlated response for increasing H (5.73 ducklings in G10 vs 4.86 in G4, and 3.09 ducklings in Pekin L-201) and maximum duration of the fertile period without increasing the embryo mortality rate. The average predicted genetic response for F was 40% of genetic standard deviation per generation of selection. The mule ducklings' feather colour also was improved. It was concluded that this study provided results for a better understanding of the genetics of the duration of fertility traits in the common female duck bred for mule and that the selection of a synthetic strain was effective method of improvement.

NMDA Receptor and NO Mediate ET-1-Induced Behavioral and Cardiovascular Effects in Periaqueductal Gray Matter of Rats

  • Ryu, Jung-Su;Shin, Chang-Yell;Yang, Sung-Jun;Lee, Tai-Sang;La, Hyun-O;Song, Hyun-Ju;Yom, Yoon-Ki;Huh, In-Hoi;Sohn, Uy-Dong
    • Archives of Pharmacal Research
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    • v.24 no.1
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    • pp.64-68
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    • 2001
  • Endothelin-1 (ET-1 ), a novel and potent vasoconstrictor in blood vessel, is known to have some functions in the rat central nervous system (CNS), In order to investigate the central functions of ET-1 , ET-1 was administered to the periaqueductal gray area (PAC) of anesthetized rats to induce barrel rolling and increase the arterial blood pressure (ABP). ET-1 had a modulatory effect on central cardiovascular and behavioral control. The selective N-methyl-D-aspartate (NMDA) receptor antagonist MK-801 (3${u}m/ol/kg$, i.p.) blocked the ET-1 induced responses, and both the nitric oxide synthase (NOS) inhibitor L-NAME (N-nitro-L-arginine mIThyl-ester 1 nmol/rat) and the nitric oxide (NO) scavenger hemoglobin (15 nmol/rat) had similar effects in redtAcing the IT-1 (10 pmol/rat)-induced behavioral changes and ABP elevation. However, NO donor sodium nitroprusside (SNP 10${u}g$, 1${u}g/rat$) decreased the ET-1 induced ABP elevation, and recovered the ET-1 -induced barrel rolling effect that was reduced by MK-801. These results suggest that ET-1 might have neuromodulatory functions such as ABP elevation and barrel rolling induction in the PAG of the rats via the NMDA receptor and NO.

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Effect of Supplemental Medicinal Plants(Artemisia, Acanthopanax and Garlic) on Growth Performance, IGF-1 and Meat Quality Characteristics in Growing-Finishing Pigs (사료내 약용 식물(인진쑥, 오가피 및 마늘)의 첨가가 육성-비육돈의 생산성, IGF-1 및 육질 특성에 미치는 영향)

  • Kwon, O.S.;Cho, J. H.;Min, B. J.;Kim, H. J.;Chen, Y. G.;Yoo, J. S.;Kim, I. H.;La, J. C.;Park, H. K.
    • Food Science of Animal Resources
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    • v.25 no.3
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    • pp.316-321
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    • 2005
  • A total of ninety six pigs ($L{\times}Y{\times}D$, 20.92(2.13kg average initial body weight) were used in a 16-week performance growth assay to determine the effect of supplemental medicinal plane (Artemisia, Acanthopanax and Garlic) on growth performance, IGF-1 of serum and carcass characteristics in finishing pigs. The dietary treatments were included 1) CON (basal diet; Control), 2) MP1 (basal diet added $0.02\%$ of medicinal plant mixtures), 3) MP2 (basal diet added $0.04\%$ of medicinal plant mixtures) and 4) MP3 (basal diet added $0.06\%$ of medicinal plant mixtures). Through entire experimental period, as medicinal plants mixture (MP) increased, there was a decrease (linear, P<0.08) in average daily feed intake and an increase (linear, P<0.02; quadratic, P<0.08) in gain/feed. The backfat thickness tended to decrease in pigs fed MP diet compared to pigs fed CON diet (linear, P<0.09; quadratic, P<0.01). Increasing medicinal plane mixture tended to increase in IGF-1 content in serum (linear, P<0.09). The hunter $a^{*}$ (redness) (linear, P<0.01) and $b^{*}$ (yellowness) (linear, P<0.02) values of longissimus muscle were affected by the dietary MP treatments. The color of longissimus muscle was higher in the dietary MP treatments than that of the muscle in the control diet (linear, P<0.03). In conclusion, the result obtained from this feeding triad suggest that the medicinal plants mixture supplementation below $0.06\%$ in diets for growing-finishing pigs can be improved growth performance, IGF-1 and meat quality.

Sex-related Clinicopathologic Differences in Patients with Adenocarcinoma of the Lung (성별에 따른 원발성 폐선암 환자들의 차이)

  • Park, Eun Ho;Jang, Tae Won;Jang, Li La;Paek, Jong yun;Oak, Chul Ho;Jung, Mann Hong;Jang, Hee Kyung
    • Tuberculosis and Respiratory Diseases
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    • v.62 no.3
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    • pp.203-210
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    • 2007
  • Background: The incidence of adenocarcinoma of the lung has been increasing worldwide, and it has been generally been accepted to be relatively unrelated to smoking with a female preponderance. The aim of this study was to examine the gender-related pathological and survival differences in patients with an adenocarcinoma of the lung. Material and Method: A retrospective review of the clinical information of patients diagnosed with an adenocarcinoma of the lung at Kosin Medical Center from January 1999 to September 2005 was performed. The patient's demographics (age, gender), smoking history, stage, serum tumor marker, pathology classification, EGFR mutation, K-ras mutation, treatment methods, and survival time were analyzed. Result: Of the 438 patients, 179 (40.9%) were female. The median age at the diagnosis was 58 years for females and 59 years for males. However, 25.8% of women and only 17.7% of men were under 50 years of age (p=0.02). The distribution of the disease stage was similar in both men and women. The bronchioloalveolar carcinoma component was diagnosed more often in women (11.2%) than in men (5.0%). The overall survival rate was higher in women than in men (p=0.01), and women had a superior therapeutic response to a combined treatment of surgery and chemotherapy. Conclusion: This study showed significant genders differences in terms of the smoking history, bronchioloalveolar carcinoma component, overall survival, and survival after combined treatment of surgery and chemotherapy. Therefore, gender differences should be considered when diagnosing and treating adenocarcinomas of the lung.

Quinacrin Induces Cytochrome c-dependent Apoptotic Signaling in Human Cervical Carcinoma Cells

  • Fasanmade, Adedigbo A.;Owuor, Edward D.;Ee, Rachel P.L.;Qato, Dima;Heller, Mark;Kong, Ah Ng Tony
    • Archives of Pharmacal Research
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    • v.24 no.2
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    • pp.126-135
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    • 2001
  • Quinacrine (QU), a phospholipase-A2 (PLA-2) inhibitor has been used clinically as a chemotherapeutic adjuvant. To understand the mechanisms leading to its chemotherapeutic effect, we have investigated QU-induced apoptotic signaling pathways in human cervical squamous carcinoma HeLa cells. In this study, we found that QU induced cytochrome c-dependent apoptotic signaling. The release of pro-apoptotic cytochrome c was QU concentration- and time-dependent, and preceded activation of caspase-9 and -3. Flow cytometric FACScan analysis using fluorescence intensities of $DiOC_6$/ demonstrated that QU-induced cytochrome c release was independent of mitochondrial permeability transition (MPT), since the concentrations of QU that induced cytochrome c release did not alter mitochondrial membrane potential (${\blacktriangle}{\Psi}_m$). Moreover, kinetic analysis of caspase activities showed that cytochrome c release led to the activation of caspase-9 and downstream death effector caspase-3, Caspase-3 inhibitor (Ac-DEVD-CHO) partially blocked QU-induced apoptosis, suggesting the importance of caspase-3 in this apoptotic signaling mechanism. Supplementation with arachidonic acid (AA) sustained caspase-3 activation induced by QU. Using inhibitors against cellular arachidonate metabolism of lipooxygenase (Nordihydroxyguaiaretic Acid, NDGA) and cyclooxygenase (5,8,11,14-Eicosatetraynoic Acid, ETYA) demonstrated that QU-induced apoptotic signaling may be dependent on its role as a PLA-2 inhibitor. Interestingly, NDCA attenuated QU-induced cytochrome c release, caspase activity as well as apoptotic cell death. The blockade of cytochrome c release by NDCA was much more effective than that attained with cyclosporin A (CsA), a MPT inhibitor. ETYA was not effective in blocking cytochrome c release, except under very high concentrations. Caspase inhibitor z-VAD blocked the release of cytochrome c suggesting that this signaling event is caspase dependent, and caspase-8 activation may be upstream of the mitochondrial events. In summary, we report that QU induced cytochrome c-dependent apoptotic signaling cascade, which may be dependent on its role as a PLA-2 inhibitor. This apoptotic mechanism induced by QU may contribute to its known chemotherapeutic effects.

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