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Antioxidative and Cellular Protective Effects of Dolwoe (Gynostemma pentaphyllum Makino) Extracts against Oxidative Stress (돌외(Gynostemma pentaphyllum Makino) 추출물의 산화적 스트레스에 대한 항산화 및 세포보호효과)

  • Kim, Kyoung Mi;Kim, A-Rang;Kim, A-Young;Ha, Ji Hun;Xuan, Song Hua;Jeong, Yoon Ju;Park, Young Min;Jeong, Hyo Jin;Hong, In Gi;Park, Soo Nam
    • Korean Journal of Pharmacognosy
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    • v.48 no.2
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    • pp.125-133
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    • 2017
  • In this study, we investigated the total phenolic and flavonoid contents, component analysis, antioxidative activity and cellular protective effects against oxidative stress on human skin cells in 50% ethanol extract, ethyl acetate fraction and aglycone fraction obtained from Gynostemma pentaphyllum (G. pentaphyllum) Makino. The DPPH (1,1-diphenyl-2-picrylhydrazyl) scavenging activites ($FSC_{50}$) of the 50% ethanol extracts, ethyl acetate fraction and aglycone fraction were 246.8, 147.2, $128.9{\mu}g/mL$, respectively. Reactive oxygen species (ROS) scavenging activities ($OSC_{50}$) of the 50% ethanol extract, ethyl acetate fraction and aglycone fraction on ROS generated in $Fe^{3+}-EDTA/H_2O_2$ system using the luminol-dependent chemiluminescence assay were 37.15, 10.74, $7.19{\mu}g/mL$, respectively. We investigated the cellular protective activity and the results showed that treatment of aglycone fraction ($0.05-0.39{\mu}g/mL$) protect human skin cells in a concentration-dependent manner when the skin cell damages were induced by treating them with $H_2O_2$. These results suggest that extract/fractions of G. pentaphyllum Makino may be applicable as natural antioxidants in cosmetics.

Characterization and Antimicrobial Activity of Lactic Acid Bacteria Isolated from Vaginas of Women of Childbearing Age (가임기 여성의 질에서 분리한 젖산 세균인 Lactobacillus plantarum UK-3의 특성 및 항균활성)

  • Ahn, Hye-Ran;So, Jae-Seong;Oh, Kye-Heon
    • Korean Journal of Microbiology
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    • v.47 no.4
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    • pp.308-315
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    • 2011
  • The purpose of this work was to examine the antimicrobial activity derived from the lactic acid bacterium, UK-3 isolated from the vaginas of women of childbearing age. Various physiological and biochemical properties of this strain were characterized. Both the BIOLOG system and phylogenetic analysis using 16S rRNA sequencing were utilized for identification, and the strain was designated as Lactobacillus plantarum UK-3, and registered in GenBank as [JK266589]. Growth rate, production of organic acids (e.g., lactic acid and acetic acid), and pH during growth were monitored. The maximum concentrations of lactic acid and acetic acid were approximately 684.11 mM and 174.26 mM, respectively, and pH changed from 7.0 to 3.7 after 72 h of incubation. High performance liquid chromatography was used to confirm lactic acid and acetic acid production. Significant antimicrobial activity of the concentrated supernatant was demonstrated against various Gram-positive (e.g., Staphylococcus aureus, Staphylococcus epidermidis, Methicillin-resistant Staphylococcus aureus, Enterococcus faecalis, Neisseria species., Listeria monocytogenes), Gram-negative bacteria (e.g., Escherichia coli, Klebsiella pneumoniae, Proteus mirabilis), and yeast (e.g., Candida albicans) by the plate diffusion method. As a result, the concentrated L. plantarum UK-3 cultures had lower acidity and inhibited the growth of all microorganisms tested, whereas the growth of L. acidophilus was not affected.

Cardiovascular Safety Pharmacology of Sibutramine

  • Yun, Jaesuk;Chung, Eunyong;Choi, Ki Hwan;Cho, Dae Hyun;Song, Yun Jeong;Han, Kyoung Moon;Cha, Hey Jin;Shin, Ji Soon;Seong, Won-Keun;Kim, Young-Hoon;Kim, Hyung Soo
    • Biomolecules & Therapeutics
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    • v.23 no.4
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    • pp.386-389
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    • 2015
  • Sibutramine is an anorectic that has been banned since 2010 due to cardiovascular safety issues. However, counterfeit drugs or slimming products that include sibutramine are still available in the market. It has been reported that illegal sibutramine-contained pharmaceutical products induce cardiovascular crisis. However, the mechanism underlying sibutramine-induced cardiovascular adverse effect has not been fully evaluated yet. In this study, we performed cardiovascular safety pharmacology studies of sibutramine systemically using by hERG channel inhibition, action potential duration, and telemetry assays. Sibutramine inhibited hERG channel current of HEK293 cells with an $IC_{50}$ of $3.92{\mu}M$ in patch clamp assay and increased the heart rate and blood pressure ($76{\Delta}bpm$ in heart rate and $51{\Delta}mmHg$ in blood pressure) in beagle dogs at a dose of 30 mg/kg (per oral), while it shortened action potential duration (at $10{\mu}M$ and $30{\mu}M$, resulted in 15% and 29% decreases in $APD_{50}$, and 9% and 17% decreases in $APD_{90}$, respectively) in the Purkinje fibers of rabbits and had no effects on the QTc interval in beagle dogs. These results suggest that sibutramine has a considerable adverse effect on the cardiovascular system and may contribute to accurate drug safety regulation.

Functional Analysis of the Stress-Inducible Soybean Calmodulin Isoform-4 (GmCaM-4) Promoter in Transgenic Tobacco Plants

  • Park, Hyeong Cheol;Kim, Man Lyang;Kang, Yun Hwan;Jeong, Jae Cheol;Cheong, Mi Sun;Choi, Wonkyun;Lee, Sang Yeol;Cho, Moo Je;Kim, Min Chul;Chung, Woo Sik;Yun, Dae-Jin
    • Molecules and Cells
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    • v.27 no.4
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    • pp.475-480
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    • 2009
  • The transcription of soybean (Glycine max) calmodulin isoform-4 (GmCaM-4) is dramatically induced within 0.5 h of exposure to pathogen or NaCl. Core cis-acting elements that regulate the expression of the GmCaM-4 gene in response to pathogen and salt stress were previously identified, between -1,207 and -1,128 bp, and between -858 and -728 bp, in the GmCaM-4 promoter. Here, we characterized the properties of the DNA-binding complexes that form at the two core cis-acting elements of the GmCaM-4 promoter in pathogen-treated nuclear extracts. We generated GUS reporter constructs harboring various deletions of approximately 1.3-kb GmCaM-4 promoter, and analyzed GUS expression in tobacco plants transformed with these constructs. The GUS expression analysis suggested that the two previously identified core regions are involved in inducing GmCaM-4 expression in the heterologous system. Finally, a transient expression assay of Arabidopsis protoplasts showed that the GmCaM-4 promoter produced greater levels of GUS activity than did the CaMV35S promoter after pathogen or NaCl treatments, suggesting that the GmCaM-4 promoter may be useful in the production of conditional gene expression systems.

Anti-fibrotic effects of Orostachys japonicus A. Berger (Crassulaceae) on hepatic stellate cells and thioacetamide-induced fibrosis in rats

  • Koppula, Sushruta;Yum, Mun-Jeong;Kim, Jin-Seoub;Shin, Gwang-Mo;Chae, Yun-Jin;Yoon, Tony;Chun, Chi-Su;Lee, Jae-Dong;Song, MinDong
    • Nutrition Research and Practice
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    • v.11 no.6
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    • pp.470-478
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    • 2017
  • BACKGROUND/OBJECTIVE: Orostachys japonicus A. Berger (Crassulaceae) has been used in traditional herbal medicines in Korea and other Asian countries to treat various diseases, including liver disorders. In the present study, the anti-fibrotic effects of O. japonicus extract (OJE) in cellular and experimental hepatofibrotic rat models were investigated. MATERIALS/METHODS: An in vitro hepatic stellate cells (HSCs) system was used to estimate cell viability, cell cycle and apoptosis by MTT assay, flow cytometry, and Annexin V-FITC/PI staining techniques, respectively. In addition, thioacetamide (TAA)-induced liver fibrosis was established in Sprague Dawley rats. Briefly, animals were divided into five groups (n = 8): Control, TAA, OJE 10 (TAA with OJE 10 mg/kg), OJE 100 (TAA with OJE 100 mg/kg) and silymarin (TAA with Silymarin 50 mg/kg). Fibrosis was induced by treatment with TAA (200 mg/kg, i.p.) twice per week for 13 weeks, while OJE and silymarin were administered orally two times per week from week 7 to 13. The fibrotic related gene expression serum biomarkers glutathione and hydroxyproline were estimated by RT-PCR and spectrophotometry, respectively, using commercial kits. RESULTS: OJE (0.5 and 0.1 mg/ mL) and silymarin (0.05 mg/mL) treatment significantly (P < 0.01 and P < 0.001) induced apoptosis (16.95% and 27.48% for OJE and 25.87% for silymarin, respectively) in HSC-T6 cells when compared with the control group (9.09%). Further, rat primary HSCs showed changes in morphology in response to OJE 0.1 mg/mL treatment. In in vivo studies, OJE (10 and 100 mg/kg) treatment significantly ameliorated TAA-induced alterations in levels of serum biomarkers, fibrotic related gene expression, glutathione, and hydroxyproline (P < 0.05-P < 0.001) and rescued the histopathological changes. CONCLUSIONS: OJE can be developed as a potential agent for the treatment of hepatofibrosis.

Effects of Salvia plebeia Herb Extracts on Anti-oxidant Activity and Whitening action (곰보배추 추출물이 항산화 및 피부미백효과에 미치는 영향)

  • Lim, Ha Na;Pyo, Young Hee;Yoon, Mi Yun
    • Journal of the Korean Applied Science and Technology
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    • v.34 no.4
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    • pp.995-1003
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    • 2017
  • Whitening and anti-oxidant effects were observed in order to investigate the biological activations of Salvia plebeia herb ethanol extracts. No toxicity was found in both B16F10 melanoma cells and Raw 264.7 cells exposed to Salvia plebeia herb ethanol extracts for 48 hour. The extracts showed significant antioxidant activity in cell-free and cell-cultured system. In the DPPH radical assay, it removed dose-dependently DPPH radicals and showed 77.6% at $100{\mu}g/mL$. In the cells, it also significantly removed silica-induced ROS generation and LPS-induced NO production in a dose dependent manner. Using L-DOPA and L-tyrosine as a substrate, tyrosinase activity was inhibited using Salvia plebeia herb ethanol extracts in a dose-dependent manner. The supression occurred to be in the B16F10 melanoma cells, where dose-dependently inhibited Salvia plebeia herb ethanol extracts of $1{\mu}g/M$ ${\alpha}$-melanocyte stimulated hormone-induced melanin production and the inhibitory effect was 30.7% at a concentration of $100{\mu}g/mL$. This suggests that the Salvia plebeia herb ethanol extracts are usable for cosmetic product developments for anti-oxidant and whitening effects.

Isolation and Characterization of Helicobacter pylori Urease Inhibitor from Rubus coreanus Miquel (복분자(Rubus coreanus Miquel)로부터 Helicobacter pylori Urease Inhibitor의 분리 및 특성)

  • 양성우;호진녕;이유현;신동훈;홍범식;조홍연
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.33 no.5
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    • pp.769-777
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    • 2004
  • A Helicobacter pylori urease inhibitor from Rubus coreanus Miquel has been isolated and partially characterized for aiming to Prevent H. pylori growth and decrease harmful accumulation of ammonia in human gastric mucosa. We screened urease inhibitory activities in 519 extracts library prepared by solvent extraction from 173 kinds of edible plants, medicinal herbs, herbs and seaweeds using a colorimetric urease assay system. As results of primary and secondary screening, 70% acetone extract of Rubus coreanus Miquel was selected as potent candidate, showing about 24% inhibitory activity. The acetone extract was sequentially partitioned into RCE/RCWI and RCB/RCW2 layers with ethyl acetate and butanol. The major active component in RCW2, water layer from butanol fractionation was revealed to be peptidic or proteinous substance by inhibitory activity determination after pronase digestion and periodate oxidation. RCW2-IIIc a was isolated by sequential column chromatography on DEAE-Toyopearl 650C, Butrl-Toyopearl 650M and Sephadex LH-20. The isolated urease inhibitor RCW2-IIIc $\alpha$, was highly pure proteinous substance with molecular weight of 13kDa by high-performance gel permeation liquid chromatography. RCW2-IIIc$\alpha$ has about 5 times higher inhibitory activity than 70% acetone extract, showing high stability against heat treatment and peptic digestion.

The Inhibitiory Effects of the Scirpi rhizoma on Melanogenesis (삼릉추출물의 멜라닌 생성 저해 효과)

  • Lee, Bum-Chun;Kim, Jin-Hwa;Sim, Gwan-Sub;Zhang, Yong-He;Pyo, Hyeong-Bae
    • Journal of the Society of Cosmetic Scientists of Korea
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    • v.31 no.4 s.54
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    • pp.305-310
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    • 2005
  • To obtain effective and safe depigmenting agents, we investigated the effects of Scirpi rhizoma, a medicine among Chinese herbs, on melanogenesis. Dried S. rhizoma was refluxed with 70% aqueous ethanol and the extract was evaporated to dryness. To determine the effects as a whitening agent, various in vitro tests were performed such as free radical scavenging activity, melanin formation assay, tyrosinase activity and expression of tyrosinase, TRP-1 and TRP-2(western blot and RT-PCR) in B16 melanoma cells. S. rhizoma showed scavenging activities of free radicals and reactive oxygen species (ROS) with the $IC_{50}\;of\;638{\mu}g/mL$ against 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical and $21.7{\mu}g/mL$ against superoxide radicals in the xanthine/xanthine oxidase system, respectively. S. rhizoma significantly inhibited melanin production in B16 melanoma cells. S. rhizoma treatment(48 h) suppressed the biosynthesis of melanin up to 27% at 100{\mu}g/mL$ and reduced tyrosinase activity up to 31% at $100{\mu}g/mL$ in B16 melanoma cells. S. rhizoma was also able to significantly inhibit tyrosinase and TRP-1 expression in protein and mRNA level. These results suggest that S. rhizoma inhibited melanin biosynthesis by regulating tyrosinase activity and expression in B16 melanoma cells. Therefore, S. rhizoma may be useful as a new antioxidant and whitening agent to inhibit melanogenesis.

Biochemical Methane Potential of Agricultural Waste Biomass (농산 바이오매스의 메탄 생산 퍼텐셜)

  • Shin, Kook-Sik;Kim, Chang-Hyun;Lee, Sang-Eun;Yoon, Young-Man
    • Korean Journal of Soil Science and Fertilizer
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    • v.44 no.5
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    • pp.903-915
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    • 2011
  • Recently, anaerobic methane production of agricultural waste biomass has received increasing attention. Until now domestic BMP (Biochemical methane potential) studies concerned with agricultural waste biomass have concentrated on the several waste biomass such as livestock manure, food waste, and sewage sludge from WWTP (Waste water treatment plant). Especially, the lack of standardization study of BMP assay method has caused the confused comprehension and interpretation in the comparison of BMP results from various researchers. Germany and USA had established the standard methods, VDI 4630 and ASTM E2170-01, for the analysis of BMP and anaerobic organic degradation, respectively. In this review, BMP was defined in the aspect of organic material represented as COD (Chemical oxygen demand) and VS (Volatile solid), and the influence of several parameters on the methane potential of the feedstock was presented. In the investigation of domestic BMP case studies, BMP results of 18 biomass species generating from agriculture and agro-industry were presented. And BMP results of crop species reported from foreign case studies were presented according to the classification system of crops such as food crop, vegetables, oil seed and specialty crop, orchards, and fodder and energy crop. This review emphasizes the urgent need for characterizing the innumerable kind of biomass by their capability on methane production.

Antimicrobial activity and characterization for defensin of synthetic oligopeptides derived from Bombus ignitus (호박벌 유래 디펜신 유전자의 분자적 특성분석 및 항균 활성)

  • Kang, Heui-Yun;Kim, In-Woo;Lee, Joon-Ha;Kwon, Young Nam;Yun, Eun-Young;Yoon, Hyung Joo;Kim, Seong-Ryul;Kim, Iksoo;Hwang, Jae-Sam
    • Journal of Sericultural and Entomological Science
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    • v.50 no.2
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    • pp.161-165
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    • 2012
  • Antimicrobial peptides of insects are found and reported as immune defence system against infectious agents. The peptides are produced by fat body cells and thrombocytoids, a blood cell type. Defensin is 38-45 amino acids long and consists of an ${\alpha}$-helix linked by a loop to an antiparallel ${\beta}$-sheet. Defensin from a bumblebee, Bombus ignitus, is known to comprise 52 amino acid residues. This peptide consists of two ${\alpha}$-helixes; ACAANCLSM and KTNFKDLWDKRF and one ${\beta}$-sheet; GGRCENGVCLCR. We carried out antibacterial activity test by radial diffusion assay against Staphylococcus aureus (Gram positive), Escherichia coli (Gram negative), Pseudomonas syringae (Gram negative), Candida albicans (fungi), MDRPA, MRSA, and VRE (antimicrobial resistant microbes) with synthetic oligopeptides from Peptron (Daejeon, Korea). The predicted curtailment fragment (GGRCEVCLCR-$NH_2$) for ${\beta}$-sheet had strong antibacterial activity when internal amino acids were removed. But, curtailment fragments (ACAANCLSM-$NH_2$ and TNFKDLWDKR-$NH_2$) of ${\alpha}$-helix were not showed antibacterial activity. These synthetic oligopeptides were showed the great activity against Gram positive and negative bacteria.