• Title/Summary/Keyword: crude extracts

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First Report of Freesia sneak virus in Freesia spp. in Korea

  • Yoon, Ju-Yeon;Choi, Youn-Jung;Choi, Gug-Seoun;Choi, Seung-Kook
    • Research in Plant Disease
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    • v.19 no.4
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    • pp.313-318
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    • 2013
  • In March, 2013, twenty symptomatic freesia plants (10 plants of cultivar Shiny Lemon and 10 plants of cultivar Shiny Gold), with striking virus-like symptoms were collected in Cheongju, Korea. The plants showed chlorotic, coalescing, interveinal, whitish, necrotic, mosaic, mottling or dark brown-to-purple necrotic spots on leaves. Freesia crude sap was directly analyzed by transmission electron microscopy, which potyvirus particles as well as long virus-like particles were detected. Total RNA extracts were analyzed for the infection of Freesia sneak virus (FreSV) by reverse transcription (RT)-PCR with primers specific to FreSV coat protein (CP) gene based on the sequences of FreSV isolates (GenBank No. GU071089, FJ807730 and DQ885455), showing 9 of 20 plants were infected. All 1305bp RT-PCR products were cloned and sequenced. Comparisons of nucleotide and deduced amino acid sequences using BLAST and bioinformatics tools resulted in 99 to 100% sequence identity with FreSV isolates FOV, Virginia, and Italy, confirming FreSV in 9 symptomatic freesia plants. Of 9 determined cDNAs of FreSV isolates, sequences of 5 cDNA clones were identical (GenBank No. AB811437) and sequences of 4 cDNA clones were identical (GenBank No. AB811792). To our knowledge, this is the first report of FreSV from Freesia spp. in Korea.

Inductions of Caspase-, MAPK- and ROS-dependent Apoptosis and Chemotherapeutic Effects Caused by an Ethanol Extract of Scutellaria barbata D. Don in Human Gastric Adenocarcinoma Cells

  • Shim, Ji Hwan;Gim, Huijin;Lee, Soojin;Kim, Byung Joo
    • Journal of Pharmacopuncture
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    • v.19 no.2
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    • pp.129-136
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    • 2016
  • Objectives: The crude extracts of Scutellaria barbata D. Don (SB) have traditionally demonstrated inhibitory effects on numerous human cancers both in vitro and in vivo. Gastric cancer is one of the most common types of cancer on world. The authors investigated the effects of an ethanol extract of Scutellaria barbata D. Don (ESB) on the growth and survival of MKN-45 cells (a human gastric adenocarcinoma cell line). Methods: The MKN-45 cells were treated with different concentrations of ESB, and cell death was examined using an MTT (3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide) assay. Analyses of sub-G1 peaks, caspase-3 and -9 activities, and mitochondrial membrane depolarizations were conducted to determine the anti-cancer effects of SB on MKN-45 cells. Also, intracellular reactive oxygen species (ROS) generation was investigated. Results: ESB inhibited the growth of MKN-45 cells, caused cell cycle arrest, and increased the sub-G1 population. In addition, ESB markedly increased mitochondrial membrane depolarization and the activities of caspase-3 and -9. ESB exerted anti-proliferative effects on MKN-45 cells by modulating the mitogen-activated protein kinase (MAPK) signaling pathway and by increasing the generation of ROS. Furthermore, combinations of anti-cancer drugs plus ESB suppressed cell growth more than treatments with an agent or ESB, and this was especially true for cisplatin, etoposide, and doxorubicin. Conclusion: ESB has a dose-dependent cytotoxic effect on MKN-45 cells and this is closely associated with the induction of apoptosis. ESB-induced apoptosis is mediated by mitochondria-, caspase- and MAPK dependent pathways. In addition, ESB enhances ROS generation and increases the chemosensitivity of MKN-45 cells. These results suggest that treatment with ESB can inhibit the proliferation and promote the apoptosis of human gastric adenocarcinoma cells by modulating the caspase-, MAPK- and ROS-dependent pathway.

Pharmacological Action of Machilus Thunbergii Siebold Zuccarini (한국산 생약제들의 혈압강하작용에 대한 연구 II. 한국산 후박수피의 혈압강하 작용)

  • Cho B.H.;Kim I.H.;Lee S.B.;Cho K.C.;Lee J.H.
    • The Korean Journal of Pharmacology
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    • v.15 no.1_2 s.25
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    • pp.45-56
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    • 1979
  • With a view to searching after a new antihypertensive or hypotensive agents in the botanical crude plants, authors intended to reevaluate several natural products caltivated in Korea. This experiment was undertaken to compare pharmacogical actions of Machilus thunbergii Siebold et Zuccarini with those of Magnolia obovata Thunberg in anesthetized rats and in normal mice. Machilus thunbergii Sieb. et Zucc., a tree belonging to the Lauraceae family, is caltivated at Ull-ung Do, and their cortecies have been used as folk medicine mingled with those of Magnolia obovata Thunberg. These two cortecies have teen also applied in chinese medicine, it was advocated that these cortecies exerted good therapeutic effects on gastritis, convulsive abdominal pain, nausea, vomiting and urinary tract disorders. Therefore, we intended to determine the pharmacological action of two palnt of different family each other, especially their effects on blood pressure and heart rate, and also their mechanism of action were observed. We studied their action with extracts of hexane(MTHE), ether(MTEE), methanol(MTME) and water(MTWE) from Machilus thunhergii Sieb. et Zucc., and also fractionations of methanol(MOME), chloroform(MOCE) and water(MOWE) from Mapolia obovata Thunberg. The results of this experiment were as follows; 1) MTME, when intravenously administered to rats, elicited the significant hypotensive responses dependent on the administered dosage. 2) MOWE was also exhibited the hypotensive effect dependent on the treated dose. 3) Depressor effect of MTME was blocked by pretreatment with hexamethonium. 4) The hypotensive response of MOWE was blocked by pretreatment with hexamethonium or hrdralazine. 5) HTME and MOWE were also observed the anticonvulsive effect and sedative effect. These results suggested that MTME may induce the hypotensive response via central sympathetic effect, but the site of action in brain are not clarified, and the hypotensive effect of MOWE may be due to dual mechanism of central sympathetic action and direct vasodilation of blood vessel.

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Biotoxic Cyanobacterial Metabolites Exhibiting Pesticidal and Mosquito Larvicidal Activities

  • Kumar, Ashok;Dhananjaya P. , Singh;Tyagi, M.B.
    • Journal of Microbiology and Biotechnology
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    • v.13 no.1
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    • pp.50-56
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    • 2003
  • A freshwater bloom-forming cyanobacterium, Microcystis aeruginosa, and local soil isolate Scytonema sp. strain BT 23 were demonstrated to contain biotoxic secondary metabolites with pesticidal and mosquito larvicidal activities. A purified toxic constituent from M aeruginosa showed an absorption maximum at 230 nm and its toxicity symptoms, Rf value on TLC, and retention time observed ill an HPLC analysis were similar to those of the hepatotoxic heptapeptide microcystin-LR. The bioactive constituent of the Scytonema sp. was less polar in nature and exhibited two peaks at 240 and 285 m. When applied to two cruciffrous pests, Pieris brassicae and Plutella flostella, the crude extracts and toxic principles from the two cyanobacteria showed significant antifeedant activity in a no-choice bioassay, and at higher concenuations exhibited contact toxicity to the insect larvae. The purified toxin from M. aeruginosa was found to be more effective and produced 97.5 and $92.8\%$ larval mortality in the two pests, fo11owing 2 h of toxin treatment at a concentration of $25{\mu}g$ Per leaf disc (2.5 cm dia.). Meanwhile, similar treatment with the purified toxin from Sytonema sp. stain BT 23 only produced 73 and $78\%$ mortality in the two pests. The cyanobacterial constituents also showed significant activity against Culex and Anopheles larvae. The M. aeruginosa toxin ($20{\mu}g\;ml^-1$) caused 98.2 and $88.1\%$ mortality in the Culex and Anopheles larvae, respectively, while the purified toxin from the Sytonema sp. was less toxic and only produced a 96.3 and $91.2\%$ mortality, respectively, at a much higher concentration ($40{\mu}g\;ml^-1$). Accordingly, the current results point to certain hitherto unknown biological properties of cyanobacterial biotoxins.

Chemical Properties and Physiological Activities of Synnemata of Beauveria bassiana

  • Yoon, Cheol-Sik;Yu, Kwang-Won;Bae, Song-Hwan;Song, Hyuk-Hwan;Park, Hyun-Soo;Lee, Chan
    • Journal of Microbiology and Biotechnology
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    • v.13 no.1
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    • pp.125-133
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    • 2003
  • Chemical properties and physiological activities of the freeze-dried synnemata of Beauveria bassiana were examined. A proximate analysis showed that the synnemata consisted mainly of carbohydrate (49.86%), protein 11.36%), and a moisture content of 30.64%. It contained a low amount of crude ash (4.76%) and fat (3.38%). The carbohydrate was composed mainly of mannose (52.3%), galactose (31.5%), glucose (13.2%), and rhamnose (3%). Trace amounts of arabinose, xylose, and fructose were present. Major amino acids In the synnemata were glutamic acid, glycine, aspartic acid, arginine, threonine, alanine, valine, leucine, lysine, and aspartic acid with the amounts of 30.42, 25.22, 17.17, 15.12, 12.65, 15.23, 12.47, 11.47, 14.24, and 17.17 mg/g, respectively. Among extracts from the synnemata, the hot-water extract showed 67% of anticomplementary activity compared to that of the positive control, followed by ethyl acetate extract (17%) and methanol extract (15%). The hot-water extract also had anticoagulant activity with 55 sec of coagulating time and this fraction exhibited the most potent Intestinal immune system modulating activity. The methanol extract showed the highest inhibitory activity (25%) on the 12-O-tetradecanoyl phorbol-13-acetate-induced superoxide ($O_2^-$) generation, followed by hot-water extract (18%) and ethyl acetate extract (10%). The data in the present study indicate that the extract of Beauveria bassiana synnemata contains some healthful chemical ingredients and it could provide beneficial physiological activities. These features of the synnemata should be of interest to the food industry as well as other industrial fields.

Simultaneous Determination of Polycyclic Aromatic Hydrocarbons and Their Nitro-derivatives in Airborne Particulates by Using Two-dimensional High-performance Liquid Chromatography with On-line Reduction and Fluorescence Detection

  • Boongla, Yaowatat;Orakij, Walaiporn;Nagaoka, Yuuki;Tang, Ning;Hayakawa, Kazuichi;Toriba, Akira
    • Asian Journal of Atmospheric Environment
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    • v.11 no.4
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    • pp.283-299
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    • 2017
  • An analytical method using high-performance liquid chromatography (HPLC) with fluorescence (FL) detection was developed for simultaneously analyzing 10 polycyclic aromatic hydrocarbons (PAHs) and 18 nitro-derivatives of PAHs (NPAHs). The two-dimensional HPLC system consists of an on-line clean-up and reduction for NPAHs in the 1st dimension, and separation of the PAHs and the reduced NPAHs and their FL detection in the 2nd dimension after column-switching. To identify an ideal clean-up column for removing sample matrix that may interfere with detection of the analytes, the characteristics of 8 reversed-phase columns were evaluated. The nitrophenylethyl (NPE)-bonded silica column was selected because of its shorter elution band and larger retention factors of the analytes due to strong dipole-dipole interactions. The amino-substituted PAHs (reduced NPAHs), PAHs and deuterated internal standards were separated on polymeric octadecyl-bonded silica (ODS) columns and by dual-channel detection within 120 min including clean-up and reduction steps. The limits of detection were 0.1-9.2 pg per injection for PAHs and 0.1-140 pg per injection for NPAHs. For validation, the method was applied to analyze crude extracts of fine particulate matter ($PM_{2.5}$) samples and achieved good analytical precision and accuracy. Moreover, the standard reference material (SRM1649b, urban dust) was analyzed by this method and the observed concentrations of PAHs and NPAHs were similar to those in previous reports. Thus, the method developed here-in has the potential to become a standard HPLC-based method, especially for NPAHs.

Chitinolytic and Chitosanolytic Activities from Crude Cellulase Extract Produced by A. niger Grown on Apple Pomace Through Koji Fermentation

  • Dhillon, Gurpreet Singh;Brar, Satinder Kaur;Kaur, Surinder;Valero, Jose R.;Verma, Mausam
    • Journal of Microbiology and Biotechnology
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    • v.21 no.12
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    • pp.1312-1321
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    • 2011
  • Enzyme extracts of cellulase [filter paper cellulase (FPase) and carboxymethyl cellulase (CMCase)], chitinase, and chitosanase produced by Aspergillus niger NRRL-567 were evaluated. The interactive effects of initial moisture and different inducers for FP cellulase and CMCase production were optimized using response surface methodology. Higher enzyme activities [FPase $79.24{\pm}4.22$ IU/gram fermented substrate (gfs) and CMCase $124.04{\pm}7.78$ IU/gfs] were achieved after 48 h fermentation in solid-state medium containing apple pomace supplemented with rice husk [1% (w/w)] under optimized conditions [pH 4.5, moisture 55% (v/w), and inducers veratryl alcohol (2 mM/kg), copper sulfate (1.5 mM/kg), and lactose 2% (w/w)] (p<0.05). Koji fermentation in trays was carried out and higher enzyme activities (FPase $96.67{\pm}4.18$ IU/gfs and CMCase $146.50{\pm}11.92$ IU/gfs) were achieved. The nonspecific chitinase and chitosanase activities of cellulase enzyme extract were analyzed using chitin and chitosan substrates with different physicochemical characteristics, such as degree of deacetylation, molecular weight, and viscosity. Higher chitinase and chitosanase activities of $70.28{\pm}3.34$ IU/gfs and $60.18{\pm}3.82$ to $64.20{\pm}4.12$ IU/gfs, respectively, were achieved. Moreover, the enzyme was stable and retained 92-94% activity even after one month. Cellulase enzyme extract obtained from A. niger with chitinolytic and chitosanolytic activities could be potentially used for making low-molecular-weight chitin and chitosan oligomers, having promising applications in biomedicine, pharmaceuticals, food, and agricultural industries, and in biocontrol formulations.

Induction of Phytoalexins by Uptake of Naphthoquinones in Cell Cultures of Petunia (Naphthoquinone류 화합물 흡수에 의한 페튜니아 배양세포내의 Phytoalexin 유도)

  • Kim, Myong-Jo;Kwak, Sang-Soo
    • Applied Biological Chemistry
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    • v.40 no.4
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    • pp.352-356
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    • 1997
  • To induce the phytoalexins in plant cell culture systems, we surveyed the antimicrobial activity following the feeding of five naphthoquinones in cell cultures of petunia. Among naphthoquinones treated, 2,5,7-trihydroxy-3-(5'-hydroxyhexyl)-1,4-naphthoquinone (3-OH NQ ) was most efficiently absorbed into the cells within 48 hr. The crude extracts of cells treated with 3-OH NQ showed a strong inhibition activity on spore germination of Aspergillus candidus $(MIC:\;32\;{\mu}g/ml)$, whereas the untreated cells showed no activity. The two active compounds, 4,2',4',${\beta}$-tetrahydroxychalcone and 4',7-dihydroxyflavone, were isolated from petunia cells treated with 3-OH NQ. The major phytoalexin, 4,2',4',${\beta}$-tetrahydroxychalcone, inhibited strongly the spore germination of A. candidus $(MIC:\;16\;{\mu}g/ml)$.

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Coenzyme $Q_{10}$ Production by Sphingomonas sp. ZUTE03 with Novel Precursors Isolated from Tobacco Waste in a Two-Phase Conversion System

  • Qiu, Lequan;Wang, Weijian;Zhong, Weihong;Zhong, Li;Fang, Jianjun;Li, Xuanzhen;Wu, Shijin;Chen, Jianmeng
    • Journal of Microbiology and Biotechnology
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    • v.21 no.5
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    • pp.494-502
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    • 2011
  • Coenzyme $Q_{10}$ ($CoQ_{10}$) is a widely used supplement in heart diseases treatment or antioxidative dietary. The microbial production of $CoQ_{10}$ was enhanced by addition of solanesol and novel precursors recovered from waste tobacco. The novel precursors were separated by silica gel and identified as ${\alpha}$-linolenic acid (LNA) and butylated hydroxytoluene (BHT) based on the effect on $CoQ_{10}$ production and GC-MS. The effects of novel precursors on $CoQ_{10}$ production by Sphingomonas sp. ZUTE03 were further evaluated in a two-phase conversion system. The precursor's combination of solanesol (70 mg/l) with BHT (30 mg/l) showed the best effect on the improvement of $CoQ_{10}$ yield. A maximal $CoQ_{10}$ productivity (9.5 mg $l^{-1}$ $h^{-1}$) was achieved after 8 h conversion, with a molar conversion rate of 92.6% and 92.4% on BHT and solanesol, respectively. The novel precursors, BHT and LNA in crude extracts from waste tobacco leaves, might become potential candidates for application in the industrial production of $CoQ_{10}$ by microbes.

Antioxidative activities of Paulownia coreana extractives (오동나무 추출물의 항산화 효능)

  • Si, Chuan-Ling;Kim, Jin-Kyu;Kwon, Dong-Joo;Bae, Young-Soo
    • Journal of Forest and Environmental Science
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    • v.22 no.1
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    • pp.13-17
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    • 2006
  • Approximately each 5 kg dried Paulownia coreana leaves, seeds, outer barks, inner barks and wood were ground, extracted with acetone-$H_2O$ (7:3. v/v). concentrated under reduced pressure and successively fractionated using n-hexane. $CH_2Cl_2$, EtOAc and $H_2O$ on a separation funnel. Each fraction was then freeze dried and weighed. The extraction yield of each sample was investigated and the yield of inner barks was the highest, while the wood was the lowest. Antioxidative activities of crude extracts, precipitates and partitioned fractions were evaluated by DPPH radical scavenging method. The results indicated that all EtOAc soluble fractions, including some of $H_2O$ soluble fractions, showed significantly high antioxidative potential compared with ${\alpha}$-tocopherol and BHT used as controls.

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