• Title/Summary/Keyword: Bio-Pharmaceutical

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JPH203, a selective L-type amino acid transporter 1 inhibitor, induces mitochondria-dependent apoptosis in Saos2 human osteosarcoma cells

  • Choi, Dae Woo;Kim, Do Kyung;Kanai, Yoshikatsu;Wempe, Michael F.;Endou, Hitoshi;Kim, Jong-Keun
    • The Korean Journal of Physiology and Pharmacology
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    • v.21 no.6
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    • pp.599-607
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    • 2017
  • Most normal cells express L-type amino acid transporter 2 (LAT2). However, L-type amino acid transporter 1 (LAT1) is highly expressed in many tumor cells and presumed to support their increased growth and proliferation. This study examined the effects of JPH203, a selective LAT1 inhibitor, on cell growth and its mechanism for cell death in Saos2 human osteosarcoma cells. FOB human osteoblastic cells and Saos2 cells expressed LAT1 and LAT2 together with their associating protein 4F2 heavy chain, but the expression of LAT2 in the Saos2 cells was especially weak. JPH203 and BCH, a non-selective L-type amino acid transporter inhibitor, potently inhibited L-leucine uptake in Saos2 cells. As expected, the intrinsic ability of JPH203 to inhibit L-leucine uptake was far more efficient than that of BCH in Saos2 cells. Likewise, JPH203 and BCH inhibited Saos2 cell growth with JPH203 being superior to BCH in this regard. Furthermore, JPH203 increased apoptosis rates and formed DNA ladder in Saos2 cells. Moreover, JPH203 activated the mitochondria-dependent apoptotic signaling pathway by upregulating pro-apoptotic factors, such as Bad, Bax, and Bak, and the active form of caspase-9, and downregulating anti-apoptotic factors, such as Bcl-2 and Bcl-xL. These results suggest that the inhibition of LAT1 activity via JPH203, which may act as a potential novel anti-cancer agent, leads to apoptosis mediated by the mitochondria-dependent intrinsic apoptotic signaling pathway by inducing the intracellular depletion of neutral amino acids essential for cell growth in Saos2 human osteosarcoma cells.

Antibacterial Activities of Ginkgo Biloba Leaves Extracts Against Isolated Bacteria from Museums (박물관에서 분리된 세균에 대한 은행잎 추출물의 항균활성)

  • Kwon, Young-Suk;Cho, Hyun-Hok;Jeong, Seong-Yun;Lee, Sang-Youb;Kim, Min-Ju;Cho, Sun-Ja;Lee, Sang-Joon
    • Journal of Environmental Science International
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    • v.15 no.10
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    • pp.983-988
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    • 2006
  • The textile remains have been affected largely by environmental factors including microorganisms because they were composed of organic compounds to be easy to damage. So, we selected 8 strains of the 131 isolated strains from museum environments and textile remains by high pretense activity, and identified them for measuring the antibacterial activity of Gingko biloba extracts. They were identified Genus Arthrobacter spp. 3 strains (Arthrobacter nicotiannae A12, Arthrobacter sp B12, Arthrobacter oxidans B13), Cenus Bacillus spp. 2 strains (Bacillus licheniformis D9, Bacillus cereus D33), Genus Pseudomonas spp. 2 strains (Pseudomonas putida A24, Pseufomonas fluorescene C21) and a Genus Staphylococcus sp. 1 strain (Staphylococcus pasteuri D3) as closest strains through the blast search of NCBI. Though antibacterial activity of the extracts of Gingko biloba leaves as MIC was lower than that of other pharmaceutical antibiotics. However the extracts was crude extracts, the extracts might have good antibacterial against most of the isolates from museum. Especially, the antifungal activity of Gingko biloba is known previously, the extracts of Gingko biloba leaves has possibility of usage as a good natural material for conservation of remains.

Purification and Characterization of a Serine Protease (CPM-2) with Fibrinolytic Activity from the Dung Beetles

  • Ahn, Mi-Young;Hahn, Bum-Soo;Ryu, Kang-Sun;Hwang, Jae-Sam;Kim, Yeong-Shik
    • Archives of Pharmacal Research
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    • v.28 no.7
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    • pp.816-822
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    • 2005
  • Catharsius protease-2 (CPM-2) was isolated from the body of dung beetles, Catharsius molossus, using a three step purification process (ammonium sulfate fractionation, gel filtration on Bio-Gel P-60, and affinity chromatography on DEAE Affi-Gel blue). The purified CPM-2, having a molecular weight of 24 kDa, was assessed homogeneously by SDS-polyacrylamide gel electrophoresis. The N-terminal amino acid sequence of CPM-2 was composed of X Val Gin Asp Phe Val Glu Glu lie Leu. CPM-2 was inactivated by $Cu^{2+}\;and\;Zn^{2+}$ and strongly inhibited by typical serine proteinase inhibitors such as TLCK, soybean trypsin inhibitor, aprotinin, benzamidine, and ${\alpha}_1$-antitrypsin. However, EDTA, EGTA, cysteine, $\beta$-mercaptoethanol, E64, and elastatinal had little effect on enzyme activity. In addition, antiplasmin and antithrombin III were not sensitive to CPM-2. Based on the results of a fibrinolytic activity test, CPM-2 readily cleaved $A{\alpha}-$ and $B{\beta}$-chains of fibrinogen and fibrin, and y-chain of fibrinogen more slowly. The nonspecific action of the enzyme resulted in extensive hydrolysis, releasing a variety of fibrinopeptides of fibrinogen and fibrin. Polyclonal antibodies of CPM-2 were reactive to the native form of antigen. The ELISA was applied to detect quantities, in nanograms, of the antigen in CPM-2 protein.

Lifespan Extension of Fermented Zizyphus jujuba Fruits in Caenorhabditis elegans (붉은 덕다리버섯 발효 대조(大棗)의 예쁜꼬마선충 수명연장효과)

  • Ji, Byeong-Uk;Park, Sung-Min;Koo, Sungtae;Lim, Byungmook;Yu, Young-Beob
    • Korean Journal of Acupuncture
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    • v.31 no.4
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    • pp.218-224
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    • 2014
  • Objectives : Zizyphus jujuba fruits(ZJF), a traditional Korean medicine has various biological activities such as anti-inflammatory, anti-oxidative and neuro-protective effects. However, it is still unclear whether ZJF has any biological effect on anti-aging. In this study, we examined the effect of ZJF on lifespan and thermal stress in C. elegans. Methods and Results : ZJF water extracts were fermented for 7 days(F7-ZJF) and 14 days(F14-ZJF) by Laetiporus sulphureus to increase secondary metabolites such as aglycone of flavonoids and terpenoids. In the lifespan assay, ZJF water extracts and fermented ZJF were treated on the agar medium plate with age synchronized egg stage of C. elegans. Treatment of F7-ZJF-$200{\mu}g/mL$ with OP-50 E. coli and F14-ZJF-$200{\mu}g/mL$ with OP-50 E. coli significantly increased life span of C. elegans(N2) at thermal stress condition of $25^{\circ}C$. Moreover mRNA levels of lifespan associated HSP 16.1, HSP 70, and HSF-1 were increased at thermal stress condition of $25^{\circ}C$. However, in the equilibration temperature of $20^{\circ}C$ after stress condition of $35^{\circ}C$ for 2 hr, F-14-ZJF-$200{\mu}g/mL$ treatment decreased the levels of heat shock protein in hsp16.2/GFP C. elegans. Conclusions : Our study indicates that prolong role of fermented-ZJF in C. elegans is mediated by control HSPs production.

Competitive Adsorption Characteristics of Cupper and Cadmium Using Biochar Derived from Phragmites communis (갈대 biochar의 구리 및 카드뮴 경쟁흡착특성)

  • Park, Jong-Hwan;Kim, Seong-Heon;Shin, Ji-Hyun;Kim, Hong Chul;Seo, Dong Cheol
    • Korean Journal of Environmental Agriculture
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    • v.34 no.1
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    • pp.21-29
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    • 2015
  • BACKGROUND: Heavy metal adsorptionnot only depends on biochar characteristics but also on the nature of the metals involved and on their competitive behavior for biochar adsorption sites. The goal of this study was to investigate the competitive absorption characteristics of Cu and Cd in mono-metal and binary-metal forms by biochar derived from Phragmites communis. METHODS AND RESULTS: Batch and column experiments were conducted to evaluate the competitive adsorption characteristics of the biocharfor Cu and Cd. In the batch experiments, the maximum adsorption capacity of Cd(63 mg/g) by biochar was higher than that for Cu (55 mg/g) in the mono-metal adsorption isotherm. On the other hand, the maximum Cu adsorption capacity (40 mg/g) by biochar was higher than that for Cd(25 mg/g) in the binary-metal adsorption isotherm. Cu was the most retained cations. Cd could be easily exchanged and substituted by Cu. The amounts of adsorbed metals in the column experiments were in the order of Cd (121 mg/g) > Cu (96 mg/g) in mono-metal conditions, and Cu (72 mg/g) > Cd (29 mg/g) in binary-metal conditions. CONCLUSION: Overall, the results demonstrated that competitive adsorption among metals increased the mobility of these metals. Particularly, Cd in binary-metal conditions lost its adsorption capacity most significantly.

Bio-Equivalence Analysis using Linear Mixed Model (선형혼합모형을 활용한 생물학적 동등성 분석)

  • An, Hyungmi;Lee, Youngjo;Yu, Kyung-Sang
    • The Korean Journal of Applied Statistics
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    • v.28 no.2
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    • pp.289-294
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    • 2015
  • Linear mixed models are commonly used in the clinical pharmaceutical studies to analyze repeated measures such as the crossover study data of bioequivalence studies. In these models, random effects describe the correlation between repeated outcomes and variance-covariance matrix explain within-subject variabilities. Bioequivalence analysis verifies whether a 90% confidence interval for geometric mean ratio of Cmax and AUC between reference drug and test drug is included in the bioequivalence margin [0.8, 1.25] performed using linear mixed models with period, sequence and treatment effects as fixed and sequence nested subject effects as random. A Levofloxacin study is referred to for an example of real data analysis.

From Bench to Market: Preparing Human Pluripotent Stem Cells Derived Cardiomyocytes for Various Applications

  • Moon, Sung-Hwan;Bae, Daekyeong;Jung, Taek-Hee;Chung, Eun-Bin;Jeong, Young-Hoon;Park, Soon-Jung;Chung, Hyung-Min
    • International Journal of Stem Cells
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    • v.10 no.1
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    • pp.1-11
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    • 2017
  • Human cardiomyocytes (CMs) cease to proliferate and remain terminally differentiated thereafter, when humans reach the mid-20s. Thus, any damages sustained by myocardium tissue are irreversible, and they require medical interventions to regain functionality. To date, new surgical procedures and drugs have been developed, albeit with limited success, to treat various heart diseases including myocardial infarction. Hence, there is a pressing need to develop more effective treatment methods to address the increasing mortality rate of the heart diseases. Functional CMs are not only an important in vitro cellular tool to model various types of heart diseases for drug development, but they are also a promising therapeutic agent for cell therapy. However, the limited proliferative capacity entails difficulties in acquiring functional CMs in the scale that is required for pathological studies and cell therapy development. Stem cells, human pluripotent stem cells (hPSCs) in particular, have been considered as an unlimited cellular source for providing functional CMs for various applications. Notable progress has already been made: the first clinical trials of hPSCs derived CMs (hPSC-CMs) for treating myocardial infarction was approved in 2015, and their potential use in disease modeling and drug discovery is being fully explored. This concise review gives an account of current development of differentiation, purification and maturation techniques for hPSC-CMs, and their application in cell therapy development and pharmaceutical industries will be discussed with the latest experimental evidence.

Active Compounds from The Aerial Parts of Polygonum multiflorum Thunberg (하수오 지상부의 활성 성분)

  • Kim, Jin-Pyo;Kim, Do-Hoon;Kim, Gi-Eun;Kim, Jin-Hyo;Chung, Sung-Hee;Kim, Sung-Gun;Whang, Wan-Kyunn
    • YAKHAK HOEJI
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    • v.50 no.6
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    • pp.351-357
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    • 2006
  • Polygoni Radix (Polygonaceae) has been used as renal disoder hematopoietic, menstrual irregularity and antiaging in Korean folk medicine. Specially aerial parts has been used for insomnia and sedative agent. In order to investigate the efficacy of antioxidant activity on these aerial parts, the bio-activity guided fraction and isolation of physiologically active substance were performed. H$_2$O, 30%, 60%, 100% MeOH were examined antioxidant activity by DPPH method. It was revealed that 30%, 60% MeOH fractions have significantly antioxidant activity. From 60% MeOH fraction, six known flavonoid glycosides (quercetin, quercetin 3-O-${\beta}$-D-galactopyranosyl-4'-O-glucopyrano side-3'-methyl ether, quercetin 3-O-${\alpha}$-L-rhannopyranoside, quercetin 3-O- ${\alpha}$-L-rhamnopyranoside-4'-methylethor, quorcetin 3-O-${\beta}$-D-galactopyranoside, quercetin 3-O-${\beta}$-D-galactopyranoside) and 3,4-Oihydrocinnarnic acid were isolated. To investigate the antioxidant activities of each com-pounds, we measured radical scavening activity with DPPH method and anti-lipid peroxidative efficacy on low density lipo-protein (LDL) with TBARS assay and anti-acetylcholinestrase activity with Ellman's Test. Four compounds (ll, IV Vl, Vll) of quercetin glycosides showed significant activity.

The Flora of Experiment Forest of Kookmin University (국민대학교 학술림의 식물상)

  • Choi, Im Jun;Lee, Jong-Won;Lim, Won Taek;Jang, Jun Ho;Kang, Shin-Ho
    • Proceedings of the Plant Resources Society of Korea Conference
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    • 2019.04a
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    • pp.45-45
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    • 2019
  • 학술림은 학생, 교수, 산림 연구기관의 실험 실습 등의 연구 기능을 하는 숲이며, 경제적, 사회적 및 환경적으로 큰 의미가 있어, 학술연구 및 교육에 있어서 중요한 필수 자산이다. 국내에 분포하는 학술림의 전체면적은 약 34,941ha로 국내 전체 면적 6,335,000ha의 0.55%를 차지하고 있다. 국내 학술림을 행정적으로 남부, 중부 및 북부 세권역으로 나눠볼 수 있는데 중부권역에 속해있는 한 곳인 국민대학교 학술림의 자생식물을 조사하였다. 국민대학교 학술림이 위치한 경상북도 안동시 길안면 배방리 일대는 청송 유네스코 세계지질공원 및 주왕산국립공원과 인접함에도 불구하고 연구가 부족한 실정이다. 본 연구에서 확인된 관속식물은 97과 292속 518종 등 총 518분류군으로 조사되었으며, 산림청지정 희귀식물로는 댕댕이나무, 산분꽃나무, 시호, 솜양지꽃, 산마늘 등 10분류군이 확인되었고, 특산식물은 청괴불나무, 참배암차즈기, 고려엉겅퀴, 분취 등 9분류군이 확인되었다. 식물구계학적 특정식물은 I등급은 고려엉겅퀴, 백선, 뻐꾹채, 초롱꽃, 투구꽃 등 26분류군, II등급은 곰취, 구와취, 노랑제비꽃, 채고추나물, 호오리새 등 11분류군, III등급은 복자기, 시닥나무, 참조팝나무, 눈개승마, 다북떡쑥 등 24분류군, Ⅳ등급은 개아마, 뚝사초, 산마늘, 왜방풍, 참배암차즈기 등 8분류군, V등급은 산분꽃나무 1분류군으로 확인되었다.

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Compound K, a ginsenoside metabolite, plays an antiinflammatory role in macrophages by targeting the AKT1-mediated signaling pathway

  • Lee, Jeong-Oog;Choi, Eunju;Shin, Kon Kuk;Hong, Yo Han;Kim, Han Gyung;Jeong, Deok;Hossain, Mohammad Amjad;Kim, Hyun Soo;Yi, Young-Su;Kim, Donghyun;Kim, Eunji;Cho, Jae Youl
    • Journal of Ginseng Research
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    • v.43 no.1
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    • pp.154-160
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    • 2019
  • Background: Compound K (CK) is an active metabolite of ginseng saponin, ginsenoside Rb1, that has been shown to have ameliorative properties in various diseases. However, its role in inflammation and the underlying mechanisms are poorly understood. In this report, the antiinflammatory role of CK was investigated in macrophage-like cells. Methods: The CK-mediated antiinflammatory mechanism was explored in RAW264.7 and HEK293 cells that were activated by lipopolysaccharide (LPS) or exhibited overexpression of known activation proteins. The mRNA levels of inflammatory genes and the activation levels of target proteins were identified by quantitative and semiquantitative reverse transcription polymerase chain reaction and Western blot analysis. Results: CK significantly inhibited the mRNA expression of inducible nitric oxide synthase and tumor necrosis factor-${\alpha}$ and morphological changes in LPS-activated RAW264.7 cells under noncytotoxic concentrations. CK downregulated the phosphorylation of AKT1, but not AKT2, in LPS-activated RAW264.7 cells. Similarly, CK reduced the AKT1 overexpression-induced expression of aldehyde oxidase 1, interleukin-$1{\beta}$, interferon-${\beta}$, and tumor necrosis factor-${\alpha}$ in a dose-dependent manner. Conclusion: Our results suggest that CK plays an antiinflammatory role during macrophage-mediated inflammatory actions by specifically targeting the AKT1-mediated signaling pathway.