• Title/Summary/Keyword: Chemical assay

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Cytotoxic and Anti-inflammatory Activities of Lipids from the Nuruk (Rhizopus oryzae KSD-815) (누룩(Rhizopus oryzae KSD-815)으로부터 분리한 지질화합물의 세포독성 및 항염증 활성)

  • Kwak, Ho-Young;Lee, Sang-Jin;Lee, Dae-Young;Bae, Nark-Hyun;Jung, La-Koon;Hong, Sung-Youl;Kim, Gye-Won;Baek, Nam-In
    • Applied Biological Chemistry
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    • v.51 no.2
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    • pp.142-147
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    • 2008
  • Nuruk is the Korean traditional Koji that contains various microorganisms and has been used to make the traditional fermented foods including alcoholic beverages. Rhizopus oryzae KSD-815 was isolated from the alcohol-fermenting Nuruk used for manufacturing traditional alcohol. In this study, the authors reported the isolation and identification of four lipids from the Nuruk (Rhizopus oryzae KSD-815) that inoculated wheat with Rhizopus oryzae KSD-815. The dried and powdered Nuruk (Rhizopus oryzae KSD-815) were extracted three times at room temperature with 80% aqueous MeOH. The extracts were partitioned with EtOAc, n-BuOH, and water, successively. The EtOAc extract was suspended in 80% MeOH and partitioned repeatedly with n-hexane. From the n-hexane fraction, four lipids were isolated through the repeated silica gel and ODS column chromatographies. According to the results of physico-chemical data including NMR, GC and MS, the chemical structures of the compounds were determined as linolenic acid methyl ester (1), palmitic acid methyl ester (2), linoleic acid (3), palmitic acid (4). Cytotoxicity was evaluated in huamn breast cancer cells, MDA-MB-231 and human hepatocarcinoma, SK-HEP-1 cells using MTT assay. Exposure of compounds 1 and 3 led to a dose-dependent inhibition of cell viability in both cancer cell lines. In addition, treatment of RAW264.7 cells with compound 3 caused inhibition of lipopolysaccharide/interferon-${\gamma}$-induced nitric oxide production.

Genetic and Physiological Discrepancies from Isolates of Sclerotinia homoeocarpa causing Zoysiagrass Dollar Spot Disease (한국잔디에 발생하는 동전마름병 원인균의 유전 및 생리적 특성차이)

  • Park, Dae-Sup;Kim, Kyung-Duck;Kihl, Joon-Yeong;Pyee, Jae-Ho
    • Asian Journal of Turfgrass Science
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    • v.20 no.1
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    • pp.65-76
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    • 2006
  • Scz1, an isolate of Sclerotinia homoeocarpa, was recently reported as a novel pathogen responsible for dollar spot disease in Zoysiagrass, a warm season turfgrass. Scz1 possessed different characteristics on mycelial pigment, mycelial affinity and host pathogenecity compared to those of Scb1, a typical isolate, obtained from creeping bentgrass, a cool season turfgrass. In this study, only three isolates, Scz1, Scz2(another analogous isolate of Sclerotinia homoeocarpa from zoysiagrass), and Scb1, were examined at the molecular level using the internal transcribed spacer(ITS) and random amplified polymorphic DNA(RAPD) assays to verify their identification and genetic variation. As a result of ITS assay, partial ITS sequences of three isolates showed 94-97% similarity with a standardized ITS sequence of S. homoeocarpa registered on BLAST. In the analysis of RAPD, range value through similarity matrix was 0.167 between Scz1 and Scb1, 0.139 between Scz2 and Scb1, and 0.713 between Scz1 and Scz2, respectively. Furthermore, tendegram analysis indicated that Scz1 and Scz2, unlike Scb1, were clustered together as accompanying a high genetic similarity. In in vitro fungicide bioassay, $EC_{50}$ value representing the sensitivity degree to propiconazole, a well-known fungicide for dollar spot disease, was 0.012 ${\mu}g/ml$ for Sczl, 0.003 ${\mu}g/ml$ for Scz2, and 0.030 ${\mu}g/ml$ for Scb1. From all data taken, we concluded that both Scz1 and Scz2 belonged to one group of S. homoeocarpa, since they exhibit the same host range and high level of genetic similarity, whereas their chemical competences to a fungicide were different. This study would provide further approach for assessing genetic diversity of S. homoeocarpa isolates as well as characterizing individual isolate against chemical exposure.

Effects of Gamma Irradiation on Color Changes and Antioxidative Activities of Caesalpinia sappan L. (감마선 조사에 의한 소목추출물의 색상변화 및 항산화 활성 평가)

  • Kwon, Hyung-Jun;Jung, U-Hee;Park, Hae-Ran;Shin, Dong-Hwa;Jo, Sung-Kee
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.36 no.8
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    • pp.1055-1061
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    • 2007
  • Caesalpinia sappan L. is an indeciduous tree distributed in China, India, Miyanmar and Vietnam. Its heartwood has long been used in oriental folk medicines to treat diseases. In this study, antioxidative activities of Caesalpinia sappan L. and the effect of gamma irradiation on its chemical and biological properties were investigated. The ethyl acetate fraction (EtOAc fr.) of Caesalpinia sappan L. was irradiated with 100 kGy of gamma ray. The dark red color of EtOAc fr. was significantly (p<0.05) removed by irradiation (Hunter L and b values increased and a value decreased). The total phenolic content of EtOAc fr. was 865 mg/g and it was increased to 1195 mg/g by gamma irradiation. DPPH radical and superoxide anion radical scavenging activities and lipid peroxidation inhibitory activity of EtOAc fr. were very high and its activities were also increased by gamma irradiation. EtOAc fr. also inhibited the irradiation-induced DNA damage of lymphocyte as determined by comet assay. In conclusion, EtOAc fr. of Caesalpinia sappan L. extract showed high antioxidative activities in vitro. Furthermore, gamma irradiation on EtOAc fr. ameliorated the color and antioxidative properties. Therefore, it can be suggested that Caesalpinia sappan L. may be a good material for antioxidant function and gamma irradiation may be applied for the improvement of chemical and biological properties of Caesalpinia sappan L.

LC-MS/MS analysis and anti-inflammatory effects of crude extract from Coptidis Rhizoma (황련 추출물의 LC-MS/MS 분석 및 항염증 효과)

  • Min-Jung, Kim;Ye-Jin, Yang;Kwang-Youn, Kim;Hun Hwan, Kim;Jae Dong, Son;Ju-Hye, Yang;Dong bin, Lee;Woo Hyun, Kim;Hu-Jang, Lee;Seon Been, Bak;Kwang-Il, Park
    • Herbal Formula Science
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    • v.31 no.1
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    • pp.1-10
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    • 2023
  • Objectives : The main aim of this study was to examine the LC-MS/MS used to identify phenolic compounds of CRE(Coptidis Rhizoma 70% EtOH Extract). Also, we investigated antioxidative activities and Anti-inflammatory activities. Methods : LC-MS/MS Analysis HPLC and LC-MS/MS were performed on a 1260 series HPLC system (Agilent Technologies, Inc., California, USA) and 3200 QTrap tandem mass system (Sciex LLC) operated in positive ion mode (spray voltage set at -4.5 kV). The solvent used was DW and Acetonitrile containing 0.1% formic acid, a gradient system was used at a flow rate of 0.5 mL/min for analysis, and a Prontosil C18 column (length, 250 mm; inner diameter, 4.6 mm; particle size, 5 ㎛; Phenomenex Co., Ltd., California, USA, Biochoff Chromatography) was used. The solvent conditions used in the mobile phases were 0-10 min at 10-15% B, 10-20 min at 20% B, 20-30 min at 25%, 30-40 min at 40%, 40-50 min at 70%, 50-60 min at 95%, and 60-70 min at 95%. The analysis was performed at a wavelength of 284 nm and a temperature of 35℃. The cell viability was measured using a 3-(4,5-dimethyethiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay and 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical scavenging activity. We examined the effects of CRE on the lipopolysaccharide (LPS)-induced production of nitric oxide (NO) in a RAW 264.7 cells Results : The chemical analysis CRE by Liquid chromatography-tandem mass spectrometry (LC-MS/MS) confirmed that Rosmarinic acid, Ferrulic acid, 3-O-feruloylquinic acid, and 5-O-feruloylquinic acid as phenolic components. DPPH radical scavenging activity was the inhibitory activity of CRE showed at 200 ㎍/mL a statistically significant level. MTT assay demonstrated that the CRE did not have a cytotoxic effect in RAW 264.7 and LPS-induced RAW264.7 cells. Also, CRE reduced NO production in RAW 264.7 cells stimulated with LPS. Conclusions : Based on these findings, The chemical analysis 4 major components CRE such as Rosmarinic acid, Ferrulic acid, 3-O-feruloylquinic acid, and 5-O-feruloylquinic acid. Moreover, we confirmed that CRE has effects antioxidant and anti-inflammatory. The results demonstrate that CRE can be used as an antioxidant and a powerful chemopreventive ingredient against inflammatory diseases.

Characteristics and Virulence Assay of Entomopathogenic Fungus Metarhizium anisopliae for the Microbial Control of Spodoptera exigua (파밤나방의 생물적 방제를 위한 곤충병원성 곰팡이 Metarhizium anisopliae의 특성 및 병원성 검정)

  • Han, Ji Hee;Kim, Hyeonggyeong;Leem, Hun Tae;Kim, Jeong Jun;Lee, SangYeob
    • The Korean Journal of Pesticide Science
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    • v.17 no.4
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    • pp.454-459
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    • 2013
  • Beet armyworm, Spodoptera exigua is difficult to control using chemical insecticides because of the fast development of insecticide resistance. For eco-friendly beet armyworm managements, various control agents are required. Entomopathogenic fungus is one of the promise control agents as an alternative to chemical control agent. We isolated entomopathogenic fungi from soil samples of Yangpyeong, Gyeonggi-do by insect-bait method using Tenebio molitor and conducted bioassay to larva of beet armyworm. The result of bioassay, a selected strain FT83 showed 100% mortality against third instar larva of S. exigua. On the basis of morphological characteristics and analysis of 18srRNA sequence for ITS, the strain FT83 was identified as a Metarhizium anisopliae. The mortality of beet armyworm showed $81.6{\pm}9.3%$ at $1{\times}10^6$ conidia/ml, 100% at $1{\times}10^7$ conidia/ml and 100% at $1{\times}10^8$ conidia/ml respectively. Therefore, we recommend to proper control efficacy against S. exigua in which more than $1{\times}10^7$ conidia/ml suspension of M. anisopliae FT83.

Simultaneous Characterization of Sofalcone and Its Metabolite in Human Plasma by Liquid Chromatography -Tandem Mass Spectrometry

  • Han, Sang-Beom;Jang, Moon-Sun;Lee, Hee-Joo;Lee, Ye-Rie;Yu, Chong-Woo;Lee, Kyung-Ryul;Kim, Ho-Hyun
    • Bulletin of the Korean Chemical Society
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    • v.26 no.5
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    • pp.729-734
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    • 2005
  • A sensitive and selective method for quantitation of sofalcone and its active metabolite in human plasma has been established using liquid chromatography-electrospray ionization tandem mass spectrometry (LC-ESI/MS/MS). Plasma samples were transferred into 96-well plate using an automated sample handling system and spiked with 10 $\mu$L of 2 $\mu$g/mL $d_3$-sofalcone and $d_3$-sofalcone metabolite solutions (internal standard), respectively. After adding 0.5 mL of acetonitrile to the 96-well plate, the plasma samples were then vortexed for 30 sec. After centrifugation, the supernatant was transferred into another 96-well plate and completely evaporated at 40 ${^{\circ}C}$ under a stream of nitrogen. Dry residues were reconstituted with mobile phase and were injected into a $C_{18}$ reversed-phase column. The limit of quantitation of sofalcone and its metabolite was 2 ng/mL, using a sample volume of 0.2 mL for analysis. The reproducibility of the method was evaluated by analyzing 10 replicates over the concentration range of 2 ng/mL to 1000 ng/mL. The validation experiments of the method have shown that the assay has good precision and accuracy. Sofalcone and its metabolite produced a protonated precursor ion ([M+H]$^+$) of m/z 451 and 453, and a corresponding product ion of m/z 315 and 317, respectively. Internal standard ($d_3$-sofalcone and $d_3$-sofalcone metabolite) produced a protonated precursor ion ([M+H]$^+$) of m/z 454 and 456 and a corresponding product ion of m/z 315 and 317, respectively. The method has been successfully applied to a pharmacokinetic study of sofalcone and its active metabolite in human plasma.

The Second-order Scattering of the Interaction of Pd Nanoparticles with Protein and Its Analytical Application

  • Guo, Xiaoyan;He, Baolin;Sun, Chuntao;Zhao, Yanxi;Huang, Tao;Liew, Kongyong;Liu, Hanfan
    • Bulletin of the Korean Chemical Society
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    • v.28 no.10
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    • pp.1746-1750
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    • 2007
  • The second-order scattering (SOS) phenomenon of the interaction of Pd nanoparticles with protein was reported and a simple, sensitive, palladium nanoparticle-based assay for trace amount of protein with SOS technique was developed. The SOS intensities were significantly enhanced due to the interaction of Pd nanoparticles with bovine serum albumin (BSA) or human serum albumin (HSA) at pH 3.5 or 4.0, respectively. The maximum SOS peak appeared at 260/520 nm (λex/λem). The optimal experiment conditions, affecting factors and the influence of some coexisting substances were checked. The SOS intensity increased proportionally with the increase of Pd concentration below 3.0 × 10?5 mol·L?1, while declined gradually above 4.0 × 10?5 mol·L?1. BSA within the range of 0.01-2.6 μg·mL?1 and HSA of 0.01-1.7 μg·mL?1 can be detected with this method and the detection limits were 2.3 and 11.2 ng·mL?1, respectively. The method was successfully applied to the quantitative detection of total protein content in human serum samples with the maximum relative standard deviation (RSD) lower than 2.6% and the recoveries over the range of 99.5-100.5%.

Effects of Exposure to Hexavalent Chromium on the Level of 8-Hydroxydeoxyguanosine and the Activities of Superoxide Dismutase and 8-Hydroxyguannine Endonuclease in Rat Lung (6가 크롬 폭로가 흰쥐 폐조직의 8-Hydroxydeoxyguanosine 농도 및 Superoxide Dismutase와 8-Hydroxyguanine Endonuclease의 활성에 미치는 영향)

  • Kim, Hun-Sik;Kim, Ro-Sa;Kim, Hyeon-Yeong;Jeong, Jae-Hwang;Kim, Heon
    • Journal of Preventive Medicine and Public Health
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    • v.32 no.1
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    • pp.101-107
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    • 1999
  • Objectives. To determine the effects of exposure to hexavalent chromium, 93 male Sprague-Dawley rats were exposed to hexavalent chromium solution. Methods. Rats were divided into 4 groups and exposed to 0.1ml of 0 mM, 0.4 mM, 2.0 mM, and 10.0 mM potassium chromate in the first experiment, and to 0.1 ml of 0 mM, 20 mM, 40 mM, and 80 mM in the second for consecutive 3 days by tracheal instillation. Three and 10 rats were the controls for the first and the second experiments, respectively. Lung tissues were then removed to measure the 8-hydroxydeoxyguanosine (8-OH-dG) level using the HPLC-ECD method, superoxide dismutase (SOD) activity using the cytochrome C method, and 8-hydroxyguanine endonuclease activity using the oligonucleotide nicking assay. Results. The results showed no significant linear relationship between chromium exposure level and 8-OH-dG level or 8-hydroxyguanine endonuclease activity. In the first experiment, 8-OH-dG level and 8-hydroxyguanine endonuclease activity increased in 0.4 mM group, and then decreased in 2.0 mM and 10.0 mM groups. The correlation coefficients between 8-OH-dG level and 8-hydroxyguanine endonuclease activity was statistically significant (P<0.01), and total SOD activity was elevated by chromium exposure in a dose-dependent manner (P<0.05). In contrast, there was no significant dose-response pattern or correlation in the secod experiment. Conclusions. Based on the fact that there was no linear relationship between chromium dose and 8-OH-dG level or activity of the repair enzyme, it seems unlikely that 8-OH-dG formation is the major mechanism of chromium carcinogenesis.

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Endoplasmic Reticulum Stress Response and Apoptosis via the CoCl2-Induced Hypoxia in Neuronal Cells (CoCl2 처리로 유도된 hypoxia상태에서 세포자살과 ER stress에 관련된 인자의 발현)

  • Kim, Seon-Hwan;Kwon, Hyon-Jo;Koh, Hyeon-Song;Song, Shi-Hun;Kwon, Ki-Sang;Kwon, O-Yu;Choi, Seung-Won
    • Journal of Life Science
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    • v.20 no.12
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    • pp.1820-1828
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    • 2010
  • Cobalt(II) chloride, a chemical compound with the formula$CoCl_2$, has been widely used in the treatment of anemia, as a chemical agent for the induction of hypoxia in cell cultures, and is known to activate hypoxic signaling. However, excessive exposure to cobalt is associated with several clinical conditions, including asthma, pneumonia, and hematological abnormalities, and can lead to tissue and cellular toxicity. It is also known to induce apoptosis. One of the questions was that of whether $CoCl_2$ might induce apoptosis via endoplasmic reticulum (ER) stress in neurons. To address this question, first, the level of DNA fragmentation was measured for assay of apoptotic rates using $CoCl_2$ with neuron PC12 cells. After confirmation of apoptosis inductions, under the same conditions, the expression levels of ER stress associated factors [ER chaperones Bip, calnexin, ERp72, ERp29, PDI, and ER membrane kinases (IRE1, ATF6, PERK)] were examined by RT-PCR and Western blotting. These results indicated that apoptosis is induced through activation of ER membrane kinases via ER stress. In conclusion, during induction of apoptosis through $CoCl_2$-induced hypoxia in neuron PC12 cells, ER membrane kinase of IRE1 was dominantly up-expressed, and, consecutively, TRAF2, which has been suggested to be one of the links connecting apoptosis and ER stress, was strongly up-expressed.

Chemical Composition and Biological Activities of Elsholtzia ciliata (Thunb.) Hylander (향유의 향기성분 분석 및 생리활성 검정)

  • Jeong, Jae-Hoon;Lim, Heung-Bin
    • Korean Journal of Medicinal Crop Science
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    • v.12 no.6
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    • pp.463-472
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    • 2004
  • This study was carried out to investigate the chemical composition of essential oils, absolutes and oleoresins isolated from Elsholtzia ciliata and the biological activities of them. Yields of essential oils, absolutes and oleoresins were 0.34%, 11.33% and 15.24%, respectively. The major component was naginate ketone in essential oils, methyl linolenate in absolutes and 9,12,15-octadecatrienoic acid in oleoresins. Eseential oils and oleoresins showed the inhibitory activities in enzyme-dependent, enzyme-independent and autooxidatve lipid peroxidation systems. $EC_{50}$ values in neutral red uptake assays 24 h of exposure times were $23.3\;{\mu}g/m{\ell}$, $341.0\;{\mu}g/m{\ell}$ and $17.2\;{\mu}g/m{\ell}$ in essential oils, absolutes and oleoresins, respectively, and essential oils and oleoresins showed the cytotoxic effect at the only high dose. Absolutes and oleoresins did not show antibiotic and mutagenic activities. On the contrary, essential oils with over $500\;{\mu}g/plate$ showed antibiotic and mutagenic activities in Ames test. Essential oils and oleoresins have a prolongating effect the ciliostasis of rat trachea.