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Induction of Nrf2/ARE-mediated cytoprotective genes by red ginseng oil through ASK1-MKK4/7-JNK and p38 MAPK signaling pathways in HepG2 cells

  • Bak, Min Ji;Truong, Van-Long;Ko, Se-Yeon;Nguyen, Xuan Ngan Giang;Jun, Mira;Hong, Soon-Gi;Lee, Jong-Won;Jeong, Woo-Sik
    • Journal of Ginseng Research
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    • v.40 no.4
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    • pp.423-430
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    • 2016
  • Background: The induction of cellular defensive genes such as phase II detoxifying and antioxidant enzymes is a highly effective strategy for protection against carcinogenesis as well as slowing cancer development. Transcription factor Nrf2 (nuclear factor E2-related factor 2) is responsible for activation of phase II enzymes induced by natural chemopreventive compounds. Methods: Red ginseng oil (RGO) was extracted using a supercritical $CO_2$ extraction system and chemical profile of RGO was investigated by GC/MS. Effects of RGO on regulation of the Nrf2/antioxidant response element (ARE) pathway were determined by ARE-luciferase assay, western blotting, and confocal microscopy. Results: The predominant components of RGO were 9,12-octadecadienoic acid (31.48%), bicyclo[10.1.0] tridec-1-ene (22.54%), and 22,23-dihydrostigmasterol (16.90%). RGO treatment significantly increased nuclear translocation of Nrf2 as well as ARE reporter gene activity, leading to upregulation of heme oxygenase-1 and NAD(P)H:quinone oxidoreductase 1. Phosphorylation of the upstream kinases such as apoptosis signal-regulating kinase (ASK)1, mitogen-activated protein kinase (MAPK) kinase (MKK)4/7, c-Jun N-terminal kinase (JNK), and p38 MAPK were enhanced by treatment with RGO. In addition, RGO-mediated Nrf2 expression and nuclear translocation was attenuated by JNK inhibitor SP600125 and p38 MAPK inhibitor SB202190. Conclusion: RGO could be used as a potential chemopreventive agent, possibly by induction of Nrf2/ARE-mediated phase II enzymes via ASK1-MKK4/7-JNK and p38 MAPK signaling pathways.

Structural characteristics of a red ginseng acidic polysaccharide rhamnogalacturonan I with immunostimulating activity from red ginseng

  • Lee, Sue Jung;In, Gyo;Han, Sung-Tai;Lee, Mi-Hyang;Lee, Jong-Won;Shin, Kwang-Soon
    • Journal of Ginseng Research
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    • v.44 no.4
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    • pp.570-579
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    • 2020
  • Background: Many researchers reported that the various immune activities of red ginseng are due to acid polysaccharides. But, the exact structural characteristics of the acidic polysaccharide in red ginseng have not been fully elucidated. Therefore, we isolated the acidic polysaccharide from red ginseng and characterized the structural property of the active moiety of this polysaccharide, which contributes to the immunostimulatory activity of red ginseng. Methods: A polysaccharide (RGP-AP-I) was purified from red ginseng via size-exclusion chromatography using Sephadex G-100. Immunostimulatary activity of RGP-AP-I was investigated via anti-complementory and macrophage stimulatory activity. The structure of RGP-AP-I was characterized by HPLC, sugar composition, β-glucosyl Yariv reagent and methylation analysis. Results: Peritoneal macrophages stimulated using RGP-AP-I significantly augmented the production of various cytokines such as interleukin (IL)-6, IL-12, and tumor necrosis factor (TNF)-α. The primary structure of RGP-AP-I was elucidated by assessing its sugar composition and methylation analysis. RGP-AP-I is a 96 kDa acidic polysaccharide, and comprises nine different monosaccharides, which mainly include sugars such as rhamnose (Rha, 9.5%), galacturonic acid (GalA, 18.4%), galactose (Gal, 30.4%), and arabinose (Ara, 35.0%). RGP-AP-I exhibited an considerable reaction with the β-glucosyl Yariv reagent, revealing the presence of arabino-β-3,6-galactan. Methylation analysis indicated that RGP-AP-I comprises 21 different glycosyl linkages, such as 3-, 4-, 6- and 3,6-linked Galp; 5-linked Araf; 2,4-linked Rhap; and 4-linked GalAp, which are characteristics of rhamnogalacturonan I (RG-I). Conclusion: we assumed that the immunostimulatory activity of RGP-AP-I may be due to the RG-I structure, which comprises a main chain with a repeating linkage unit, [→2)-Rhap-(1→4)-GalAp-(1→] and three groups of side chains such as (1→5)-linked arabinan, (1→4)-linked galactan, and arabino-β-3,6-galactan, which branch at the C(O)4 positions of Rha residues in the main chain of RGP-AP-I.

New Species of the Genus Metschnikowia Isolated from Flowers in Indonesia, Metschnikowia cibodasensis sp. nov.

  • Sjamsuridzal, Wellyzar;Oetari, Ariyanti;Nakashima, Chiharu;Kanti, Atit;Saraswati, Rasti;Widyastuti, Yantyati;Ando, Katsuhiko
    • Journal of Microbiology and Biotechnology
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    • v.23 no.7
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    • pp.905-912
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    • 2013
  • A novel species, Metschnikowia cibodasensis, is proposed to accommodate eight strains (ID03-$0093^T$, ID03-0094, ID03-0095, ID03-0096, ID03-0097, ID03-0098, ID03-0099, and ID03-0109) isolated from flowers of Saurauia pendula, Berberis nepalensis, and Brunfelsia americana in Cibodas Botanical Garden, West Java, Indonesia. The type strain of M. cibodasensis is ID03-$0093^T$ (= NBRC $101693^T$ =UICC $Y-335^T$ = BTCC-$Y25^T$). The common features of M. cibodasensis are a spherical to ellipsoidopedunculate shaped ascus, which contains one or two needle-shaped ascospores, and lyse at maturity. Asci generally develop directly from vegetative cells but sometimes from chlamydospores. The neighbor-joining tree based on the D1/D2 domain of nuclear large subunit (nLSU) ribosomal DNA sequences strongly supports that M. cibodasensis (eight strains) and its closest teleomorphic species, M. reukaufii, are different species by a 100% bootstrap value. The type strain of M. cibodasensis, ID03-$0093^T$, differed from M. reukaufii NBRC $1679^T$ by six nt (five substitutions and one deletion) in their D1/D2 region of nLSU rDNA, and by 18 nt (five deletions, four insertions, and nine substitutions) in their internal transcribed spacer regions of rDNA, respectively. Four strains representative of M. cibodasensis (ID03-$0093^T$, ID03-0095, ID03-0096, and ID03-0099) showed a mol% G+C content of $44.05{\pm}0.25%$, whereas that of M. reukaufii NBRC $1679^T$ was 41.3%. The low value of DNA-DNA homology (5-16%) in four strains of M. cibodasensis and M. reukaufii NBRC $1679^T$ strongly supported that these strains represent a distinct species.

Human and Animal Study on the Natural Food for Obesity and Metabolic Syndrome Risk Factors (비만 및 대사성증후군 위험인자에 대한 천연물 식품의 인체 및 동물 효능연구)

  • 문근아;최선미;김선형;김성수;강지연;윤유식
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.32 no.8
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    • pp.1394-1400
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    • 2003
  • In this study a natural composition containing oriental herbs, KSH28, for reducing obesity and metabolic syndrome was constructed and its efficacy was evaluated in animal and human. To investigate the anti-obesity effect of KSH28, animal study was conducted using high fat diet-induced obese mice. KSH28 significantly decreased body weight and adipose tissue in high fat diet-fed obese mice. The mean size of fat cells in adipose tissue was significantly reduced. Glucose and triglyceride levels were also significantly decreased. To elucidate its efficacy in human, a natural food containing KSH28 with grains, vegetables, vitamins, minerals and dietary fibers was constructed and 40 subjects (8 male and 32 female) were tested for the change of body composition, blood pressure and blood lipid profile. All subjects had 2 pack (309 each) of natural food per day for 4 weeks. Compared to the baseline value, body fat was significantly reduced, however, water, protein and mineral contents in the body were not changed, suggesting selective reduction of fat tissue. Blood pressure and serum lipid profile were significantly decreased to reduce risk for metabolic syndrome. Serum GPT, a liver function indicator, was not changed and no significant side effects were detected. Therefore, it was shown that the KSH28 is a safe and effective composition for reducing obesity and metabolic syndrome.

A Potential-Based Panel Method for the Analysis of A Two-Dimensional Super-Cavitating Hydrofoil (양력판(揚力板) 이론(理論)에 의(依)한 2차원(次元) 수중익(水中翼)의 초월(超越) 공동(空洞) 문제(問題) 해석(解析))

  • Y.G. Kim;C.S. Lee;J.T. Lee
    • Journal of the Society of Naval Architects of Korea
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    • v.28 no.2
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    • pp.159-173
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    • 1991
  • This paper describes a potential-based panel method formulated for the analysis of a super-cavitating two-dimensional hydrofoil. The method employs normal dipoles and sources distributed on the foil and cavity surfaces to represent the potential flow around the cavitating hydrofoil. The kinematic boundary condition on the wetted portion of the foil surface is satisfied by requiring that the total potential vanish in the fictitious inner flow region of the foil, and the dynamic boundary condition on the cavity surface is satisfied by requiring thats the potential vary linearly, i.e., the tangential velocity be constant. Green's theorem then results in a potential-based integral equation rather than the usual velocity-based formulation of Hess & Smith type. With the singularities distributed on the exact hydrofoil surface, the pressure distributions are predicted with improved accuracy compared to those of the linearized lilting surface theory, especially near the leading edge. The theory then predicts the cavity shape and cavitation number for an assumed cavity length. To improve the accuracy, the sources and dipoles on the cavity surface are moved to the newly computed cavity surface, where the boundary conditions are satisfied again. This iteration process is repeated until the results are converged. Characteristics of iteration and discretization of the present numerical method are much faster and more stable than the existing nonlinear theories. The theory shows good correlations with the existing theories and experimental results for the super-cavitating flow. In the region of small angles of attack, the present prediction shows and excellent comparison with the Geurst's linear theory. For the long cavity, the method recovers the trends of the Wu's nonlinear theory. In the intermediate regions of the short super-cavitation, the method compares very well with the experimental results of Parkin and also those of Silberman.

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Preliminary Report of Use-Effectiveness of Ovulation Method in Korea (자연피임법으로서의 배란법의 피임효과)

  • Bae, S.C.;Chung, Y.J.;Rha, J.G.;Oh, W.S.;Kim, S.J.
    • Clinical and Experimental Reproductive Medicine
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    • v.3 no.1
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    • pp.27-32
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    • 1976
  • In recent years, explosive increase in population has been damaging mankind in terms of deprivation of natural resources and more of economical demands. Therefore, we have thought about problems as to counter-balance the increasing population, and reached a resolution of artificial methods of controlling birth. In the past, though now used in some of contraceptive methods, extravaginal ejaculation, condom were commonly used. But recently, pills, IUD and several kinds of operative procedures are quite popular. Though the recent methods are known to be effective compared with the traditional methods, a certain unwanted side effects as well as limited value of usages now must be discussed. On this aspect, we are trying to research for a ideal methods such as symptom free, more of natural way of family planning and try to zero the failure rate. And also it has been suggested that only if those scientific methods of controlling birth can be base on religious concepts of moral being, it will be enlightened. At the Happy Family Planning Clinic of St. Mary's Hospital, we apply Billing's ovulation method to out-patients who want contraception and usually advise them to use it on a self-care basis. For a retrospective study of the presurvey data analysis and use-effectiveness of the ovulation method from April 1, 1975 to Nov., 30, 1975, we have dealt with a total of 1,383 women (urban areas 465, rural areas 918). The results of preliminary survey were as follows; 1) Among 465 women in Seoul areas, the failure rate was 10.3, which signified unplanned pregnancies of 32 women. 27 of the 32 women were pregnant due to the users own failure and the remaing 5 due to the failure of the method. Therefore, the failure of the method accounted for 1.6. 2) Among 918 women in rural areas, the failure rate was 15.2 signifing unplanned pregnancies in 93 women. The cause of the failure in 81 of the 93 women was attributable to the user's own mistake and that in the remaing 12 to the default of the method. Therefore, the failure attributable to the method accounted for 2.0.

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Effect of Interlayer on TiN and CrN Thin Films of STS 420 Hybrid-Deposited by AlP and DC Magnetron Sputtering (AIP 와 스퍼터링으로 복합증착된 420 스테인리스강의 TiN과 CrN 박막에 미치는 중간층의 영향)

  • Choi, Woong-Sub;Kim, Hyun-Seung;Park, Burm-Su;Lee, Kyung-Ku;Lee, Doh-Jae;Lee, Kwang-Min
    • Korean Journal of Materials Research
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    • v.17 no.5
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    • pp.256-262
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    • 2007
  • Effects of interlayer and the combination of different coating methods on the mechanical and corrosion behaviors of TiN and CrN coated on 420 stainless steel have been studied. STS 420 specimen were tempered at $300^{\circ}C$ for 1 hr in vacuum furnace. The TiN and CrN thin film with 2 ${\mu}m$ thickness were coated by arc ion plating and DC magnetron sputtering following the formation of interlayer for pure titanium and chromium with 0.2 ${\mu}m$ thickness. The microstructure and surface analysis of the specimen were conducted by using SEM, XRD and roughness tester. Mechanical properties such as hardness and adhesion also were examined. XRD patterns of TiN thin films showed that preferred TiN (111) orientation was observed. The peaks of CrN (111) and $Cr_2N$ (300) were only observed in CrN thin films deposited by arc ion plating. Both TiN and CrN deposited by arc ion plating had the higher adhesion and hardness compared to those formed by magnetron sputtering. The specimen of TiN and CrN on which interlayer deposited by magnetron sputtering and thin film deposited by arc ion plating had the highest adhesion with 22.2 N and 19.2 N. respectively. TiN and CrN samples shown the most noble corrosion potentials when the interlayers were deposited by using magnetron sputtering and the metal nitrides were deposited by using arc ion plating. The most noble corrosion potentials of TiN and CrN were found to be approximately -170 and -70 mV, respectively.

Microarray Analysis of Gene Expression Affected by Water-extracts of Pinelliae rhizoma in a Hypoxic Model of Cultured Rat Cortical Cells (배양대뇌신경세포 저산소증모델에서 반하여 의한 유전자표현의 변화)

  • Kwon, Gun-Rok;Jung, Hyun-Jung;Shin, Gil-Jo;Moon, Il-Soo;Lee, Won-Chul;Jung, Seung-Hyun
    • Journal of Life Science
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    • v.19 no.7
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    • pp.905-916
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    • 2009
  • Pinelliae rhizoma (Pr, 半夏) is a traditional medicine used in the treatment of incipient stroke. We investigated the effects of Pr on gene expression in a hypoxic model using cultured rat cortical cells. Pr (2.5 $\mu$g/ml) was added to the culture medium on DIV 12. A hypoxic shock (2% 0$_2$/5% CO$_2$, 37$^{\circ}$C, 3 hr) was given two days later (on DIV 14), and total mRNAs were isolated at 24 hr post-shock from both Pr-treated samples and untreated control cultures. Microarray using TwinChip $^{TM}$ Rat-5K (Digital Genomics, Seoul) indicated that Pr upregulated genes for cell growth and differentiation (tubb5, tgfa, ptpn11, n-ras, pdgfa) and antiapoptosis (mcl-1), while downregulating the apoptosis-induced gene (tieg). Therefore, it is interpreted that Pr protects neurons from hypxoic shock by maintaining cell growth and differentiation and by preventing apoptosis.

Effect of Supplementation of Trehalose, Glycerol on Conventional Freezing and Vitrification of Boar Sperm

  • Choi, Sun-Ho;Lee, Mi-Jin;Lee, Kyung-Mi;Sa, Soo-Jin;Kim, Hyun-Jong;Jin, Hyun-Ju;Song, Yong-Sup;Park, Jun-Cheol
    • Journal of Embryo Transfer
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    • v.29 no.4
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    • pp.397-401
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    • 2014
  • The boar sperm has more lipid droplets and specialty of seminal plasma compared with other species, causing difficulties of freezing sperm and decreases for the utilization of frozen semen into the artificial insemination. However, several studies reported significant results for the recovery of sperm motility and reproductive by addition of cryoprotectants and seminal plasma after thawing. This study was designed to investigate the effects of supplementation of trehalose or glycerol in the LEY (lactose and egg yolk in BTS) solution for the conventional freezing and vitrification process. Two boars aged 16 months were used to collect semen for 2 times in a week. The samples were allotted to 3 freezing solutions (LEY + glycerol 10.5% + OEP 1.5%, LEY + trehalose 1M + OEP 1.5%, and sucrose 1.5M + trehalose 1 M + OEP 1.5%) after centrifugation at 800 g for 10 minutes. Semen was equilibrated in freezing solutions for 10 minutes and injected into plastic straws with 2~3 air bubbles to minimize freezing damages. Vitrification was performed to locate sperm in 5 cm above $LN_2$ for 5 minutes, and the conventional freezing was conducted with an automatic freezer. Motility and survival rates were measured by CASA (Computer assisted sperm an alyzing system) and FITC (Fluorescein isothiocyanate), respectively after thawing semen at $50^{\circ}C$ for 12 seconds. The results were analyzed by ANOVA with STATVIEW statistical program. The vitrificatioin solution (LEY + 10.5% glycerol + 1.5% OEP) presented higher motility (20.9%) than other solutions while the solution (LEY + 1M trehalose + 1.5% OEP) showed the lowest (motility : 5.2%). However, survival rates of vitrified sperms detected by FITC showed 1~4% live sperms in almost of dead sperms at all vitrification solutions' groups, but survival rate of freezing solution of LEY + 1M trehalose + 1.5% OEP LEY and LEY + 10.5% glycerol + 1.5% OEP were showed 49%, and 79%, respectively. There were differences (P<0.05) survival rate of conventional freezing in LEY + 10.5% glycerol + 1.5% OEP and LEY + 1M trehalose + 1.5% OEP and the remaining showed no differences. The results suggested that vitrified boar semen was not enough to be utilized for the artificial insemination, but it showed possibility to utilize for ICSI and conventional freezing with glycerol would be useful method for artificial insemination in pig while we choose the outstanding semen against tolerance to freezing damages.

Antimelanogenic Effect and Whitening of Anthocyanin Rich Fraction from Seeds of Liriope platyphylla (맥문동 종실 안토시아닌 분획물의 멜라닌 생성 억제 및 미백 효과)

  • Choung, Myoung Gun;Hwang, Young Sun;Kim, Gi Ppeum;Ahn, Kyung Geun;Shim, Hoon Seob;Hong, Seung Beom;Choi, Jae Hoo;Yu, Chang Yeon;Chung, Ill Min;Kim, Seung Hyun;Lim, Jung Dae
    • Korean Journal of Medicinal Crop Science
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    • v.21 no.5
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    • pp.361-371
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    • 2013
  • This study was performed to determine the antimelanogenic effect and tyrosinase inhibitory activities of anthocyanin rich fraction (AN-SLP) from Liriope platyphylla Wang et Tang seeds. Anthocyanins isolated from L. platyphylla seeds revealed the presence of four major anthocyanin components, which were tentatively identified as delphinidin-3-Oglucoside, delphinidin-3-O-rutinoside, petunidin-3-O-rutinoside, and malvidin-3-O-rutinoside using semipreparative HPLC, $^1H$-NMR, $^{13}C$ NMR, FAB-MS and LC/ES-MS. The inhibitory effect of AN-SLP on tyrosinase activity was studied using in vitro (against mushroom tyrosinase) and ex vivo (against B16 melanoma cell tyrosinase) models. Cellular tyrosinase activity was decreased by AN-SLP treatment in B 16 melanoma cells through dose dependent manner, but AN-SLP did not inhibit mushroom tyrosinase and L-DOPA oxidation directly. AN-SLP showed melanin inhibition by 53.2% at 50 ${\mu}g/m{\ell}$ which was 0.7 times more efficient than the antimelanogenic effect of commercial arbutin and kojic acid (36.5%) also did not show cell toxicity. Additionally, AN-SLP inhibited the activity of ${\alpha}$-glucosidase and the glycosylation of tyrosinase in melanoma cell. The resulting unsaturated glycosylation of tyrosinase makes it unstable and disturb correct transportation. From theses results, we conclude that AN-SLP could be used as anti-melanogenic agent for skin whitening.