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Shikonin Induces Apoptotic Cell Death via Regulation of p53 and Nrf2 in AGS Human Stomach Carcinoma Cells

  • Ko, Hyeonseok;Kim, Sun-Joong;Shim, So Hee;Chang, HyoIhl;Ha, Chang Hoon
    • Biomolecules & Therapeutics
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    • v.24 no.5
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    • pp.501-509
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    • 2016
  • Shikonin, which derives from Lithospermum erythrorhizon, has been traditionally used against a variety of diseases, including cancer, in Eastern Asia. Here we determined that shikonin inhibits proliferation of gastric cancer cells by inducing apoptosis. Shikonin's biological activity was validated by observing cell viability, caspase 3 activity, reactive oxygen species (ROS) generation, and apoptotic marker expressions in AGS stomach cancer cells. The concentration range of shikonin was 35-250 nM with the incubation time of 6 h. Protein levels of Nrf2 and p53 were evaluated by western blotting and confirmed by real-time PCR. Our results revealed that shikonin induced the generation of ROS as well as caspase 3-dependent apoptosis. c-Jun-N-terminal kinases (JNK) activity was significantly elevated in shikonin-treated cells, thereby linking JNK to apoptosis. Furthermore, our results revealed that shikonin induced p53 expression but repressed Nrf2 expression. Moreover, our results suggested that there may be a co-regulation between p53 and Nrf2, in which transfection with siNrf2 induced the p53 expression. We demonstrated for the first time that shikonin activated cell apoptosis in AGS cells via caspase 3- and JNK-dependent pathways, as well as through the p53-Nrf2 mediated signal pathway. Our study validates in partly the contribution of shikonin as a new therapeutic approaches/agent for cancer chemotherapy.

A Novel Bioassay System for Screening of Compounds Affecting Anthocyanin Biosynthesis Pathway in White Corn Leaf Segment (백화옥수수 잎절편을 이용한 안토시아닌 생합성 조절제 탐색용 신규 검정법 확립)

  • Kim, Jin-Seog;Lee, Byung-Hoi;Lee, Jung-Ae;Oh, Kwang-Hoon;Cho, Kwang-Yun
    • Journal of Plant Biotechnology
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    • v.30 no.2
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    • pp.207-214
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    • 2003
  • This study was carried out to establish a novel bioassay system for screening of compounds affecting aromatic amino acid or anthocyanin biosynthesis through investigating a degree of sucrose-induced anthocyanin formation such as size of plant material, buffer conditions, light intensity and irradiated duration, incubation temp., etc were determined and standard procedure (suitable experimental condition) was set up as follows. The second leaf blade of white corn seeding induced by fluridone treatment were segmented into a size of 5${\times}$5 min. The segments were floated on the solution of 1% sucrose in 1.0mM MES buffer (pH6.0∼6.5) and incubated at 26$^{\circ}C$ for 2days under the continuous light condition(70∼100$\mu$mol m$^{-2}$ s$^{-1}$ ). Anthocyanin in the purpled tissues was extracted with methanol containing 1% HCl and the optical density of the clear supematants was determined at 528mm. Influences of some chemicals were tested using this system. Glyphosate, 5-enolpyruvylshikimate 3-phosphate synthase inhibitor, showed most sensitive response with I$_{50}$ value at 3.3$\mu$M. Dicyclohexylcarbodiimide(DCCD) and parachloromercuribenzenesulfonic acid(PCMBS) had a relatively strong ingibition with I50 value at 7.1$\mu$M and 10.2$\mu$M, respectively. These results show that sucrose-induced anthocyanin formation in white com leaf segment provide a very simple and rapid system for searching new compounds affecting aromatic amino acid or anteocyanin biosynthesis by screening at less than 10$\mu$M.

Isolation and Quantitative Analysis of BACE1 Inhibitory Compounds from Cirsium maackii Flower

  • Bhatarrai, Grishma;Seong, Su Hui;Jung, Hyun Ah;Choi, Jae Sue
    • Natural Product Sciences
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    • v.25 no.4
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    • pp.326-333
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    • 2019
  • The purpose of our study was to evaluate anti-AD potential of Cirsium maackii flowers. MeOH extract, CH2Cl2, EtOAc, and n-BuOH fraction of this flower notably inhibited BACE1 (IC50 = 76.47 ± 1.66, 22.98 ± 1.45, 8.65 ± 0.63, and 72.47 ± 3.04 ㎍/mL, respectively). β-amyrenone (49.70 mg) (1), lupeol acetate (1.43 g) (2), lupeol (1.22 g) (3), lupenone (23.70 mg) (4), β-sitosterol (1.01 g) (6), and β-sitosterol glucoside (13.00 mg) (7) from CH2Cl2, apigenin (100.20 mg) (8), luteolin (19.00 mg) (9), apigenin 7-O-glucuronide methyl ester (21.30 mg) (14), and tracheloside (53.70 mg) (5) from EtOAc, apigenin 5-O-glucoside (11.00 mg) (10), luteolin 5-O-glucoside (11.00 mg) (11) and apigenin 7-O-glucuronide (91.00 mg) (12) from n-BuOH, and luteolin 7-O-glucuronide (22.00 mg) (13) from H2O fraction were isolated. HPLC showed high levels of 8, 9 and 12 in MeOH extract (33.07 ± 0.07, 31. 44 ± 0.17 and 16.89 ± 0.33 mg/g, respectively), EtOAc (161.01 ± 1.78, 96.93 ± 0.34 and 73.38 ± 0.06 mg/g, respectively), and n-BuOH fraction (32.18 ± 0.33, 44.31 ± 0.32 and 105.94 ± 0.36 mg/g, respectively). Since, 3 and 9 are well-known BACE1 inhibitors, the anti-AD activity of C. maackii flower might be attributable to their presence.

Preparation of Jeung-Pyun Added with Ultrafiltred Powder of Sunmul (순물의 한외여과(ultrafiltration)막 분리농축분말을 이용한 증편의 제조)

  • Chung, Hai-Jung;Joo, Sin-Youn;Kim, Woo-Jung
    • Korean journal of food and cookery science
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    • v.21 no.5
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    • pp.647-654
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    • 2005
  • This study was conducted to investigate the quality characteristics of Jeung-Pyun prepared with the addition of ultrafiltered(UF) sunmul powder. Jeung-Pyun was prepared with five different levels of UF powder(0%, 0.5%, 1%, 2% and 3%) and the physico-chemical properties were examined. The pH of Jeung-Pyun batters decreased from $4.57{\sim}5.69$ to $4.38{\sim}5.03$ during 4 hours of fermentation at $35^{\circ}C$. The weight of Jeung-Pyun decreased, whereas the volume increased, with increasing UF powder content. Total isoflavone content was $0.41{\sim}2.39\;mg%$ before cooking and increased to $0.77 {\sim}3.80\;mg%$ after cooking. The increase of aglycones was particularly observed after cooking. The incorporation of UF powder in Jeung-Pyun lowered the lightness values but increased the redness and yellowness values. Rheology test showed that hardness was decreased with increasing UF powder amount. The values of gumminess and brittleness were the highest in the control group and decreased with increasing UF powder amount. Scanning electron microscope showed that the size of the air cells decreased with increasing UF powder levels. The results of sensory evaluation showed that there were no significant differences in scores of sweetness and moistness among the samples. Therefore, Jeung-Pyun prepared with UF to 3% addition of UF powder was considered to be as acceptable as Jeung-Pyun prepared without UF power.

Water desalination by membrane distillation using PVDF-HFP hollow fiber membranes

  • Garcia-Payo, M.C.;Essalhi, M.;Khayet, M.;Garcia-Fernandez, L.;Charfi, K.;Arafat, H.
    • Membrane and Water Treatment
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    • v.1 no.3
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    • pp.215-230
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    • 2010
  • Poly(vinylidene fluoride-co-hexafluoropropylene), PVDF-HFP, hollow fiber membranes were prepared by the dry/wet spinning technique using different polyethylene glycol (PEG) concentrations as non-solvent additive in the dope solution. Two different PEG concentrations (3 and 5 wt.%). The morphology and structural characteristics of the hollow fiber membranes were studied by means of optical microscopy, scanning electron microscopy, atomic force microscopy (AFM) and void volume fraction. The experimental permeate flux and the salt (NaCl) rejection factor were determined using direct contact membrane distillation (DCMD) process. An increase of the PEG content in the spinning solution resulted in a faster coagulation of the PVDF-HFP copolymer and a transition of the cross-section internal layer structure from a sponge-type structure to a finger-type structure. Pore size, nodule size and roughness parameters of both the internal and external hollow fiber surfaces were determined by AFM. It was observed that both the pore size and roughness of the internal surface of the hollow fibers enhanced with increasing the PEG concentration, whereas no change was observed at the outer surface. The void volume fraction increased with the increase of the PEG content in the spinning solution resulting in a higher DCMD flux and a smaller salt rejection factor.

A Simulation Study on the Annual Heating Performance of the Seawater-Source Screw Heat Pump (해수열원 스크류 히트펌프의 연간 난방운전 성능 모사)

  • Baik, Young-Jin;Kim, Min-Sung;Chang, Ki-Chang;Lee, Young-Soo;Kim, Hyeon-Ju
    • Journal of the Korean Solar Energy Society
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    • v.32 no.3
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    • pp.88-95
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    • 2012
  • In this study, in order to utilize the seawater as a heat source at Gangneung city near the East Sea in Korea, an annual heating performance of a screw heat pump was simulated. For a simulation, the maximum heating capacity of heat pump was assumed at 3.5 MW. An ambient temperature at Gangneung city was calculated from the TMY2 weather data, while the seawater temperature was calculated from the regression equation based on the measurement by the National Fisheries Research and Development Institute of Korea. The heating load was assumed linearly dependent on the ambient temperature, while the maximum heating load was assumed to appear when the ambient temperature is below $-2.4^{\circ}C$, which is the temperature of TAC 2.5% for heating at Gangneung city. A heat pump performance at full-load was calculated from the regression equation, which involves refrigerant's evaporating and condensing temperatures, based on a commercial screw compressor performance map. A heating supply temperature which determines refrigerant's condensing temperature was assumed linearly dependent on the heating load. A performance degradation due to the part-load operation of heat pump was also considered. Simulation results show that an annual heating coefficient of performance ($COP_H$) of a seawater-source screw heat pump is approximately 2.8 and that it is necessary to improve part-load performance to increase an annual performance of the heat pump.

A Study on Quality Characteristics of Bokbunja-Pyun Added with Rubi Fruit Juice (복분자 착즙액을 첨가한 복분자편의 품질 특성에 관한 연구)

  • Han Sook-Kyoung;Yang Hyang-Sook;Rho Jeong-Ok
    • Journal of the East Asian Society of Dietary Life
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    • v.16 no.3
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    • pp.371-376
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    • 2006
  • The purpose of this study was to investigate the quality characteristics of Bokbunja-Pyun, using various levels of Rubi fruit juice (0%, 4%, 6% and 8%). The chemical composition and physical characteristics of Bokbunja and Bokbunja-Pyun were analyzed and sensory evaluation was conducted. The statistical data analyses were completed using the SPSS program. The chemical components of Bokbunja were moisture 89.2%, sweetness 9.8 Brix%, pH 3.77, citric acid 8.98%, and vitamin C 26mg. The L-value of Bokbunja-Pyun was significantly decreased (p<0.001), but a- and b-values were increased with increasing Rubi fruit juice content (p<0.001). Among the mechanical characteristic, the hardness was the highest on R2 ($470{\pm}82.67$), the adhesiveness on R3 ($-5.00{\pm}1.0$), the cohesiveness on R4 ($99.22{\pm}6.95$) and the gumminess on R1 ($167.9{\pm}49.8$). The springiness was notsignificantly different between these groups. From the sensory evaluation, appearance, hardness, and flavor showed no significant variation, but transparency, elasticity, and overall quality were significantly different between the test groups. It was concluded that the optimum content of added of Rubi fruit juice was 6%, in proportion to the total weight.

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Alkaloids from the Tuber of Aconitum chiisanense (지리바꽃 괴경의 알카로이드)

  • Lee, Moo-Taek;Sung, Hwan-Kil;Whang, Wan-Kyunn;Kim, Il-Hyuk
    • YAKHAK HOEJI
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    • v.41 no.2
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    • pp.161-173
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    • 1997
  • Tuber of Aconitum chiisanense(Ranunculaceae) a specific medicinal plant in Korea, which is known to have the activity to recover reduced metabolism of feeble patients and has been used to symptoms such as pain, paralysis, atonia and coldness of extremities, etc. were studied. The powdered tubers of the plant were extracted with 10% EtOH 3 times and the combined extract was dissolved in 1N HCl solution and washed with ethyl acetate. The aqueous layer was basified with solid $Na_2CO_3$ and extracted with $CHCl_3$ to obtain an alkaloidal fraction. The alkaloidal fraction was subjected to column chromatography using silica gel, alumina and Sephdex LH 20, etc. From the alkaloidal fraction, five diterpene alkaloids, mesaconitine, aconitine, hypaconitine, 8-O-ethyl 14-benzoylmesaconine and talatizamine, were isolated and identified on the basis of their physico-chemical properties and spectroscopic evidences($^1H$-, $^{13}C$-NMR, EI-MS, IR, 2D-NMR) respectively. Especially the Compound IV, 8-O-ethyl 14-benzoylmesaconine, was assumed to be an artifact resulting from mesaconitine during extraction procedures. The contents of mesaconitine, aconitine and hypaconitine in the mother tuber of this plant were 0.300%, 0.024%, and 0.068%. And that of the attached tuber(new one) of this plant were 0.336%, 0.034% and 0.240% respectively.

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Secondary Ion Man Spectrometry: Theory rind Applications in Geosciences (이차이온질량분석기의 원리와 지질학적 응용)

  • 최변각
    • The Journal of the Petrological Society of Korea
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    • v.10 no.3
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    • pp.222-232
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    • 2001
  • Secondary ion mass spectrometry (SIMS) uses focused high-speed primary ions to produce secondary ions from sample surface that are analyzed through a mass filter. SIMS is often called as ion microprobe, since it offers a micrometer-scale spatial resolution. Although the precision and accuracy of SIMS are not as good as many conventional mass spectrometers, it has several advantages such as small sample-size requirement, high spatial resolution and capability of in-situ analysis. In the field of geochemistry/cosmochemistry, SIMS is widely used for (1) stable isotope geochemistry of H, C, O, S, etc., (2) geochronology of U/Th-bearing minerals, (3) lateral distribution of trace elements in a mineral, and (4) discovery of presolar grains and investigation of their isotopic compositions.

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Characterization of a QTL associated with chlorophyll content using progeny from an interspecific cross in rice (Oryza Sativa L.)

  • Shim, Kyu-Chan;Luong, Ngoc Ha;Kim, Sun Ha;Jeon, Yun-A;Lu, Xin;Ahn, Sang-Nag
    • Proceedings of the Korean Society of Crop Science Conference
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    • 2017.06a
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    • pp.23-23
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    • 2017
  • Rice (Oryza sativa L.) is the world's most important cereal crop. In crop plant, chlorophyll content and leaf senescence could affect grain filling and yield. We analyzed a QTL associated with chlorophyll content and delayed leaf senescence using high chlorophyll near isogenic line (HC-NIL). HC-NIL derived from a cross between Oryza sativa cv. Hwaseong as a recurrent parent and wild species O. grandiglumis as a donor parent showed higher chlorophyll content than Hwaseong. To identify QTL associated with chlorophyll content, 58 $F_3$ and 38 $F_4$ lines were developed from a cross between HC-NIL and Hwaseong. For QTL analysis, simple sequence repeat (SSR) markers were used for genotyping and one-way ANOVA was conducted. A QTL for chlorophyll content (qCC2) was detected in chromosome 2 and explained 24.63% of phenotypic variation. The senescence effect of the qCC2 was examined in dark-induced incubation (DII). Detached leaves from Hwaseong and HC-NIL were incubated on 3mM MES buffer (pH 5.8) at $27^{\circ}C$ under complete dark condition. After 3 days of incubation, the Hwaseong leaves turned yellow, but the HC-NIL leaves were green. HC-NIL has higher chlorophyll content with delayed senescence than Hwaseong. These results indicated that qCC2 is associated with stay-green phenotype. To know whether the qCC2 is responsible for leaf functionality, ion leakage test and Fv/Fm measurement were performed. Both experiment results showed that differences were observed between Hwaseong and HC-NIL but it was not statistically significant. These results might suggest that the qCC2 is possibly related to chlorophyll content and non-functional stay-green phenotype.

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