• Title/Summary/Keyword: crude extracts

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Use of Gelatin Particle Agglutination Test for the Detection of Cymbidium mosaic virus in Cattleya Plants

  • Han, Jung-Heon;Jeong, Hyoo-Won;La, Yong-Joon
    • The Plant Pathology Journal
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    • v.17 no.6
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    • pp.325-328
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    • 2001
  • Gelatin particle agglutination test (GPAT) was used to detect Cymbidum mosaic virus (CymMV) in Cattleya plants. Gelatin particles were coated with purified anti-CymMV immunoglobulin of 25-100 $\mu\textrm{g}$/ml and were subjected to several different concentrations of purified CyMfV as well as varying dilutions of orchid leaf extracts. The GPAT detected purified CymMV up to a minimum concentration of 10 $\mu\textrm{g}$/ml. CymMV was detected from crude sap extract of infected Cattleya leaves and roots up to 1:51,200 and 1:25,600 dilutions, respectively. However, the optimum range of leaf and root sap dilutions was between 50-100. Non-specific reactions were not encountered from any of the healthy orchid plants tested. The entire GPAT process was completed within 2-3 hours. This test was found to be very useful for the detection of CymMV in orchids because it is sensitive, economical, and easy to perform.

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Changes in Chemical Composition and Biological Activities of , Oriental Crude Drugs by Food. Processing Techniques (II) - Changes in Paeonol Contents in Roasted Moutan Cortex - (식품학적 가공에 의한 생약의 성분 및 활성 변화 (II) - Roasting 처리에 의한 목단피 (Moutan Cortex) 에탄올 추출물 중 Paeonol의 함량변화 -)

  • Jeon, So-Young;Kim, Eun-Kyung;Kwak, Hye-Min;Kim, Ja-Young;Lim, Jung-Hyun;Chung, Shin-Kyo;Song, Kyung-Sik
    • Korean Journal of Pharmacognosy
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    • v.35 no.4 s.139
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    • pp.388-392
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    • 2004
  • The paeonol content in roasted Moutan Cortex was increased about three times compared to that of untreated one. The paeonol content reached at its maximum level $(454.3\;{\mu}g/mg\;ethanol\;extract)$ after roasting at $190^{\circ}C$ for 30 minutes. Roasting processing did not affect on the DPPH radical scavenging activity of Moutan extracts.

Isolation and Numerical Identification of Streptomyces humidus strain S5-55 Antagonistic to Plant Pathogenic Fungi

  • Lim, Song-Won;Kim, Jeong-Dong;Kim, Biom-Seok;Hwang, Byung-Kook
    • The Plant Pathology Journal
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    • v.16 no.4
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    • pp.189-199
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    • 2000
  • To search for the antifungal substances, various actino-mycete isolates were obtained from various soils of Korea using plate dilution method on the humic acid vitamin agar plates. In the screening procedures using a dual culture method, 32 actionomycete isolates were selected, which showed the inhibitory activity against mycelial growth of plant pathogenic fungi Altirnaria mali, Colletotrichum gloeosporides, Fusarium oxysporum f.sp. cucumerinum, Magnaporthe grisea, Phytophthora capsici, and Rhizoctonia solani. Bioassay of the crude extracts from culture filtrates and mycelial mets revealed that 12 antagonistic actionomycetes produced highly active antifungal substances. Actinomycete strain S5-55 which showed the substantial antifungal activity against the tested fungi was selected for production of the antifungal substances. Based on the cytochemical and morphological characteristics, strain S5-55 was identified as a Streptomyces species. The results of the numerical identification using the TAXON program confirmed that Streptomyces strain S5-55 was identical with Streptomyces humidus including in TAXON major cluster 19. The production of antifungal substance was most favorable when S. humidus strain S5-55 was cultivated for 10 dats on soluble starch broth supplemented with $K_2$HPO$_4$. The antifungal substances active against the plant pathogenic fungi P. capsici and M. grisea were partially purified using $\textrm{C}_{18}$ reversed-phase column chromatography.

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Peroxynitrite-scavenging Activity of the Halophyte Limonium tetragonum (염생식물 갯질경이의 Peroxynitrite 소거 활성)

  • Lee, Jung-Im;Kong, Chang-Suk;Jung, Myoung-Eun;Hong, Joo-Wan;Noh, Il;Seo, Young-Wan
    • Ocean and Polar Research
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    • v.33 no.2
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    • pp.185-191
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    • 2011
  • Crude extracts of Limonium tetragonum and their solvent-partitioned fractions were evaluated for their potential to scavenge authentic $ONOO^-$, and $ONOO^-$ derived from 3-morpholinosydnonimine (SIN-1). Four flavonol glycosides (1-4) were isolated by activity-guided separation. Their chemical structures were elucidated by extensive 2 D NMR experiments and by comparison with published spectral data. These compounds were also estimated for their peroxynitrite scavenging effects. The scavenging ratios of compounds 1-4 on authentic $ONOO^-$ were 56, 37, 56, and 54%, respectively, at a concentration of 1 ${\mu}M$. On the other hand, the inbihition ratios of compounds 1-4 against $ONOO^-$ generation from SIN-1 were 59, 39, 44, and 54% at the same concentration, respectively.

In vitro Evidence that Purified Yeast Rad27 and Dna2 are not Stably Associated with Each Other Suggests that an Additional Protein(s) is Required for a Complex Formation

  • Bae, Sung-Ho;Seo, Yeon-Soo
    • BMB Reports
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    • v.33 no.2
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    • pp.155-161
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    • 2000
  • The saccharomyces cerevisiae Rad27, a structure-specific endonuclease for the okazaski fragment maturation has been known to interact genetically and biochemically with Dna2, an essential enzyme for DNA replication. In an attempt to define the significance of the interaction between the two enzymes, we expressed and purified both Dna2 and Rad27 proteins. In this report, Rad27 could not form a complex with Dna2 in the three different analyses. The analyses included glycerol gradient sedimentation, protein-column chromatography, and coinfection of baculoviruses followed by affinity purification. This is in striking contrast to the previous results that used crude extracts. These results suggest that the interaction between the two proteins is not sufficiently stable or indirect, and thus requires an additional protein(s) in order for Rad27 and Dna2 to form a stable physical complex. This result is consistent with our genetic findings that Schizosaccharomyces pombe Dna2 is capable of interacting with several proteins that include two subunits of polymerase $\delta$, DNA ligase I, as well as Fen-1. In addition, we found that the N-terminal modification of Rad27 abolished its enzymatic activity. Thus, as suspected, we found that on the basis of the structure determination, N-terminal methionine indeed plays an important role in the nucleolytic cleavage reaction.

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Study on Anti-thrombotic Activities of KIOM2003-080 (KIOM2003-080 처방에 대한 항혈전 효능 연구)

  • Jeon, Won-Kyung;Kim, Ho-Kyoung;Lee, Ju-Hyun;Ko, Byoung-Seob;Lee, A-Yeong;Lee, Kyung-Goo;Shin, Hyeun-Kyoo
    • Korean Journal of Oriental Medicine
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    • v.13 no.2 s.20
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    • pp.143-148
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    • 2007
  • The present study examined inhibitory effects of 20 efficient experience prescriptions on platelet aggregation induced by collagen in human whole blood using the impedance method of aggregometry. Among them, a hot water extract of KIOM 2003-080 was selected to be the most effective candidate. In an in vivo study using a mouse acute thrombosis model, the anti-thrombotic effects of the KIOM2003-080 crude extract were also observed. In addition, we accessed bio-marker of platelet activation using thromboxane B2 by ELISA assay. A significantly decrease in thromboxane B2 production was seen in the presence of KIOM2003-080. Consequently, the results from this experiment provide pharmacological evidence for the traditional use of KIOM2003-080 prescription, suggesting that its hot water extracts could be used to prevent platelet aggregation and thrombosis disease.

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Purification and Characterization of Antioxidant Substance from the Stem Bark of Rhus verniciflua (옻나무 껍질에서 항산화물질의 정제와 특성)

  • Kim, Jung-Bae
    • The Korean Journal of Food And Nutrition
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    • v.14 no.6
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    • pp.527-531
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    • 2001
  • In order to isolate antioxidant substances from Rhus verniciflua (RV) , the dried stem bark was extracted with water. The crude water extracts was purified by using HPLC method with a DEAE (anionic type) , CN and ODS column. The purified compound remained stable at pH 3.0∼6.0. but unstable above pH 6.5. It was stable at 100$^{\circ}C$ for 4 hours, but still had about 80% of residual activity after treatment at 100$^{\circ}C$ for 5 hours. In antimicrobial test, no inhibition was observed against Gram-positive and negative bacteria. This compound was stronger than that of commercial antioxidant showed that DPPH test, such as BHT, BHC at the same concentration (20 $\mu\textrm{g}$/ml) .

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Hepatoprotective and Antioxidative Effects of Alisma orientale

  • Rhew, Kwang-Yul;Choi, Hyuck-Jai;Kim, Nam-Jae;Lee, Jang-Hoon
    • Natural Product Sciences
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    • v.17 no.4
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    • pp.285-290
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    • 2011
  • The rhizome of Alisma orientale Juzep (Alismataceae) has been used as a crude drug for diabetes, edema, inflammation and urinary disturbances in oriental medicine. Recent animal studies have shown that the extract of Alisma orientale rhizome (AOR) can potently lower high levels of serum lipids and improve insulin resistance, which are usually detected in patients and animals with non-alcoholic fatty liver disease. So, we studied the antioxidative effects of AOR extracts and fraction in vitro and their protective effects against acute hepatotoxicity induced by $CCl_4$ in vivo.. We then investigated the effects of each fraction on hepatotoxicity induced by tert-butyl hydroperoxide (t-BHP). DAOR (dichloromethane fraction of the Alisma orientale rhizome) scavenged free radicals and superoxide anions. DAOR protected against $CCl_4$ induced hepatotoxicity. DAOR had hepatoprotective and antioxidative effects against t-BHP-induced hepatotoxicity in HepG2 cells and in rats.

Effects of the Combined-Preparation of Crude Drugs on Pentobarbital-induced Sleeping Time (복합한약제제가 Pentobarbital에 의해 유도된 수면시간에 미치는 영향)

  • Han, Young Taek;Kim, Dae Keun;Eun, Jae Soon
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.27 no.6
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    • pp.759-763
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    • 2013
  • This experiment was performed to investigate whether 50% ethanol extracts of the combined-preparation of Longanae Arilus, Chrysanthemi Flos, Zizyphi Fructus and Ginseng Radix alba (CPE) has hypnotic effects and/or enhances pentobarbital-induced sleeping time. Locomotor activity was evaluated using a ambulometer of tilting-type. The sedative-hypnotic effects were evaluated by measuring the sleeping onset time and sleeping time in pentobarbital-treated mice 30 min. after oral administration of CPE and muscimol. The intracellular $Cl^-$ concentration of cerebellar granule cells was estimated using $Cl^-$ sensitive fluorescence probe N-(ethoxycarbonylmethyl)-6-methoxyquinolinium (MQAE). CPE (150 mg/kg) decreased the locomotor activity, but CPE itself did not induce sleep. However, CPE reduced sleeping onset and prolonged sleeping time induced by pentobarbital (42 mg/kg). In addition, CPE (2 ${\mu}g/ml$) and pentobarbital (2.5 ${\mu}M$) itself did not affect on the chloride influx in primary cultured cerebellar granule cells, but the combination of CPE and pentobarbital (2.5 ${\mu}M$) increased the chloride influx onto the cells. In conclusion, it is suggested that CPE might augment pentobarbital-induced sleep through the increase of chloride influx.

A Spectrophotometric Assay for Cytochrome P450 Monooxygenase Activity

  • Lee, Sung-Eun;Choi, Won-Sik;Park, Byeoung-Soo;Lee, Byung-Ho
    • Applied Biological Chemistry
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    • v.41 no.4
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    • pp.213-217
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    • 1998
  • An assay for cytochrome P450 monooxygenase activity by determination of the products of organophosphate oxidation via inhibition of acetylcholinesterase was described. Extracts from strains of Oryzaephilus surinamensis selected for resistance to chlorpyrifos-methyl (QVOS 102), fenitrothion (VOS F) and malathion (VOS 3), and a standard susceptible strain VOS 48, were incubated with an organophosphate in the presence of a NADPH-generating system and acetylcholinesterase. The degree of inhibition of the acetylchoinesterase activity was converted to manooxygenase activity using standard curves for the inhibition of acetylcholiesterase by chlorpyrifos-methyl-oxon, fenitrooxon and malaoxan. Activity was detectable in VOS 48 and was present at different increased levels with the different organophosphates in the three resistant strains, suggesting that different forms of P450 might be involved in organophosphate oxidation in these insects. The assays were carried out using crude insect homogenates and much smaller samples of insect material than the standard aldrin to dieldrin assay. It should be possible to use the method for determination of monooxygenase activity in single insert.

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