• Title/Summary/Keyword: In vitro screening

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Screening of Arachidonic Acid Cascade Related Enzymes Inhibitors from Korean Indigenous Plants (2) (한국 자생식물로부터 아라키돈산 대사계 효소 저해제 검색 (2))

  • 정혜진;문태철;이은경;손건호;김현표;강삼식;배기환;안인파;권동렬
    • YAKHAK HOEJI
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    • v.47 no.2
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    • pp.69-77
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    • 2003
  • Arachidonic acid (AA), which is stored in membrane glycerophospholipids, is liberated by phospholipase $A_2$ (PLA$_2$) enzymes and is sequentially converted to cyclooxygenases (COXs) and lipoxygenases (LOXs) then to various bioactive PGs, and LTs. In order to find the specific inhibitors of AA metabolism especially PLA$_2$, COX-2, 5-LO and lyso PAF acetyltransferase, 120 Korean residential plants extracts were evaluated for their inhibitory activity on PGD$_2$, LTC$_4$ production from cytokine-induced mouse bone marrow-derived mast cells (BMMC) and arachidonic acid released from phospholipid and PAF production from lyso PAF. From this screening procedure, methanol extract of ten indigenous plant such as Salix gracilistyla, Sedum kamtschaticum, Cirsium chanroenicum, Hypericum ascyron, Astilbe chinensis, Agrimonia pilosa, Aristolochia manshuriensis, Vodia daniellii, Pyrola japonica, Styrax obassia were found to inhibit production of inflammatory mediators in vitro assay system.

Antifungal activities of coumarins isolated from Angelica gigas and Angelica dahurica against Plant pathogenic fungi (당귀와 백지로부터 분리한 Coumarin계 물질들의 식물병원균에 대한 항균활성)

  • Ryu, Shi-Yong;Kim, Young-Sup;Kim, Heung-Tae;Kim, Seong-Ki;Choi, Gyung-Ja;Kim, Jeoung-Seob;Lee, Seon-Woo;Heor, Jung-Hee;Cho, Kwang-Yun;Kim, Jin-Cheol
    • The Korean Journal of Pesticide Science
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    • v.5 no.3
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    • pp.26-35
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    • 2001
  • In order to search potent antifungal substances from domestic plants, 40 plants cultivated in Korea were collected. After extracting with methanol (MeOH) and concentrating to dryness, the MeOH extracts were screened for in vivo antifungal activity against six plant diseases at a concentration of $2000{\mu}g/mL$. Fourteen extracts showed disease-controlling activity more than 90% against at least one of the 6 plant diseases tested; eight, seven, and three extracts controlled more than 90% the development of rice blast, tomato late blight, and wheat leaf rust, respectively. However, none of the extracts exhibited in vivo antifungal activity more than 90% against rice sheath blight, tomato gray mold, and barley powdery mildew. From the MeOH extracts of Angelica gigas and A. dahurica showing potent controlling activity against rice blast, 1 and 2 antifungal substances, respectively, were isolated by solvent partitioning and column chromatography. The three compounds were identified to be coumarins, namely, decursin, imperatorin, and isoimperatorin, by mass spectrometry and NMR spectroscopy. They were examined for in vitro and in vivo antifungal activities together with umbelliferone (7-bydroxycournarin) and scopoletin (6-methoxy-7-hydroxycoumarin) containing a free hydroxyl group at position 7 to investigate the structure-activity relationship. In vitro, most of 50% growth inhibitory concentrations ($IC_{50}$) were over $200{\mu}g/mL$, indicating that they have relatively weak antifungal activity. The antifungal activity of decursin and scopoletin, containing cyclic alkoxy groups instead of free hydroxyl group at position 7, was stronger than umbelliferone and scopoletin. Especially, decursin and imperatorin showed potent antifungal activities against Pythium ultimum and Magnaporthe grisea, respectively, with $IC_{50}$ values less than $25{\mu}g/mL$. In vivo, decursin and imperatorin showed potent antifungal activity against rice blast, whereas other coumarins hardly controlled the development of 6 plant diseases tested. These results suggest that the antifungal activity of 7-hydroxycoumarin derivative is substantially increased when the hydroxyl group at position 7 is protected by a stable cyclic alkoxy grouping.

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The Emergence of Behavioral Testing of Fishes to Measure Toxicological Effects

  • Brooks, Janie S.
    • Toxicological Research
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    • v.25 no.1
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    • pp.9-15
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    • 2009
  • Historically, research in toxicology has utilized non-human mammalian species, particularly rats and mice, to study in vivo the effects of toxic exposure on physiology and behavior. However, ethical considerations and the overwhelming increase in the number of chemicals to be screened has led to a shift away from in vivo work. The decline in in vivo experimentation has been accompanied by an increase in alternative methods for detecting and predicting detrimental effects: in vitro experimentation and in silico modeling. Yet, these new methodologies can not replace the need for in vivo work on animal physiology and behavior. The development of new, non-mammalian model systems shows great promise in restoring our ability to use behavioral endpoints in toxicological testing. Of these systems, the zebrafish, Danio rerio, is the model organism for which we are accumulating enough knowledge in vivo, in vitro, and in silico to enable us to develop a comprehensive, high-throughput toxicology screening system.

Synthesis and In Vitro Cytotoxicity of Cinnamaldehydes to Hyman Solid Tumor Cells

  • Kwon, Byoung-Mog;Lee, Seung-Ho;Choi, Sang-Un;Park, Sung-Hee;Lee, Chong-Ock;Cho, Young-Kwon;Sung, Nack-Do;Bok, Song-Hae
    • Archives of Pharmacal Research
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    • v.21 no.2
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    • pp.147-152
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    • 1998
  • Cinnamaldehydes and related compounds were synthesized from various cinnamic acids based on the $2^{I}$-hydroxycinnamaidehyde isolated from the bark of Cinnamomum cassia Blume. The cytotoxicity to human solid tumor cells such as A549, SK-OV-3, SK-MEL-2, XF498 and HCT15 were measured. Cinnamic acid, cinnamates and cinnamyl alcohols did not show any cytotoxicity against the human tumor cells. Cinnamaldehydes and realted compounds were resistant to A549 cell line up to 15 .mu.g/ml. In contrast, HCT15 and SK-MEL-2 cells were much sensitive to these cinnamaidehyde analogues which showed $ED{50} values 0.63-8.1{\mu}g/ml.$Cytotoxicity of the saturated aldehydes was much weak compared to their unsaturated aldehydes. From these studies, it was found that the key functional group of the cinnamaldehyde-related compounds in the antitumor activity is the propenal group.p.

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Development of In Vitro Seedling Screening Method for Selection of Resistant Rice Against Bakanae Disease (벼 키다리병 저항성 검정을 위한 기내 유묘 검정법 개발)

  • Lee, Yong-Hwan;Lee, Myeong-Ji;Choi, Hyo-Won;Kim, Sung-Taek;Park, Jin-Woo;Myung, Inn-Shik;Park, Kyung-Seok;Lee, Se-Weon
    • Research in Plant Disease
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    • v.17 no.3
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    • pp.288-294
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    • 2011
  • Rice bakanae disease, caused by the fungus Fusarium fujikuroi, is one of the most important rice diseases and distributed widely in Asia. Resistance screening system in rice field had been established. However, the evaluation results of the system vary according to the environmental conditions when the test is conducted. To develop precise and rapid evaluation method of disease resistance of rice to bakanae disease, in vitro screening system was attempted in this study. The six cultivars namely, 'Nampyeongbyeo', 'Junambyeo', 'Chucheongbyeo', 'Samcheonbyeo', 'Odaebyeo' and 'Hwasinbyeo' were tested. They were planted onto MS agar medium (10 ml) in test tube ($450\;mm{\times}{\phi}30\;mm$) and incubated at $25^{\circ}C$ and $28^{\circ}C$ in growth chamber under 12 hr light condition. Symptoms of over growth appeared a few days after seeding and then seedling were withered 2-3 weeks after over growth. The disease symptoms such as leaf dryness on top of rice were appeared in the 'Nampyeongbyeo' from 28 days at the concentration of $10^5$ spores/ml culturing at $28^{\circ}C$ and then withered completely 35 days after seeding. Whereas the other varieties withered entirely 19-23 days after seeding. Using the in vitro seedling screening method, 72 rice varieties were investigated to select resistant cultivar. Finally, two resistant cultivars ('Nampyeongbyeo' and 'Inwolbyeo') and seven moderately resistant cultivars ('Hwadongbyeo', 'Seokjeongbyeo', 'Samgwangbyeo', 'Sampyeongbyeo', 'Nonghobyeo', 'Heukjinjubyeo' and, 'Joanbyeo') were selected. If in vitro seedling screening method was used for evaluation of bakanae disease resistance, it would be completed within 35 days after sowing of rice seed.

Synthesis, Spectral and Antimicrobial Studies of Some N(2)-Substituted Tetrahydroindazoles

  • Amirthaganesan, Shanmugasundaram;Aridoss, Gopalakrishnan;Park, Keun-Soo;Lim, Kwon-Taek;Jeong, Yeon-Tae
    • Bulletin of the Korean Chemical Society
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    • v.31 no.5
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    • pp.1135-1142
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    • 2010
  • A series of N(2)-benzothiazolyl substituted tetrahydroindazoles has been synthesized via cyclic ${\beta}$ keto esters. Optimum reaction condition was found as acidic toluene and effect of higher acidity towards substituted hydrazines in situ was described. Synthesized compounds have been achieved as single isomer and characterized by using 1D and 2D NMR spectral reports. Antimicrobial screening was carried out for the synthesized compounds along with a series of N(2)-pyridyl tetrahydroindazoles.1 The results of the in vitro antimicrobial screening studies revealed that compounds 13, 16 against Staphylococcus aureus, 11 against Escherichia coli, 10-12, 16 against Pseudomonas aeruginosa and 12 against Klebsiella pneumoniae recorded almost two-fold better activity compared to the standard drug used.

Antifungal Activity of Bacillus sp. Against Phytophthora infestans

  • Kim Hye-Sook;Yi Yong-Sub;Choi Gyung-Ja;Cho Kwang-Yun;Lim Yoong-Ho
    • Journal of Microbiology and Biotechnology
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    • v.16 no.3
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    • pp.487-490
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    • 2006
  • Because of consumer rejection of chemical pesticides and the appearance of microorganisms that are resistant to fungicides, we tried to discover biopesticides. Of 13 microorganisms isolated from Shrimp-jeotkal, a Bacillus sp. showed strong activity against tomato late blight caused by Phytophthora infestans. Its activity was tested both in vivo and in vitro. The identification of the strain was carried out based on 16S rDNA analysis and the morphology by scanning electron microscopy.

HIV Integrase Inhibitory Activity of Agastache regosa

  • Kim, Hye-Kyung;Lee, Hyeong-Kyu;Shin, Cha-Gyun;Huh, Hoon
    • Archives of Pharmacal Research
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    • v.22 no.5
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    • pp.520-523
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    • 1999
  • We have been screening anti-HIV integrase compounds from Korean medicinal plants by using an in vitro assay system which is mainly composed of recombinant human immunodeficency virus type 1 integrase and radiolabeled oligonucleotides. From the above screening, the aqueous methanolic extract of the roots of Agastache rugosa exhibited a significant activity. Bioactivity-guided chromatographic fractionation of the methanolic extract resulted in the isolation of rosmarinic acid. The structure of the compound was determined by spectroscopic data and by the comparison with the reported values. The $IC_{50}$ of the rosmarinic acid was approximately $10{\mu}g/ml$ against HIV integrase.

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In vitro Plant Propagation: A Review

  • Kumar, Nitish;Reddy, M.P.
    • Journal of Forest and Environmental Science
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    • v.27 no.2
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    • pp.61-72
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    • 2011
  • Micropropagation is an alternative mean of propagation that can be employed in mass multiplication of plants in relatively shorter time. Recent modern techniques of propagation have been developed which could facilitate large scale production of true-to-type plants and for the improvement of the species using genetic engineering techniques in the next century. An overview on the in vitro propagation via meristem culture, regeneration via organogenesis and somatic embryogenesis is presented. The usefulness of the plants in commercial industry as well as propagation techniques, screening for various useful characteristics and the influence of different cultural conditions in the multiplication, rooting and acclimatization phases on the growth of tissue cultured plant discussed.

Cloning of $\beta$-Tubulin Gene and Effect of Pencycuron on Tubulin Assembly in Rhizoctonia solani

  • Roh, Seong-Hwan;Heung-Tae KIm;Isamu-Yamaguchi
    • The Plant Pathology Journal
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    • v.15 no.1
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    • pp.68-71
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    • 1999
  • To illustrate the action mechanism of pencycuron on Rhizoctonia solani, two experiments were conducted including the comparison of amino acids of $\beta$-tubulin between R-C (sensitive isolate) and Rh-131 (non-sensitive isolate), and the inhibitory effect of pencycuron on tubulin assembly in vitro. Both $\beta$-tubulin genes of R-C and Rh-131 proved to have 1,582 nucleotides encoding a protein of 445 amino acids, showing 98% homology in amino acid sequences between them. It was found that codons at 103, 236, and 267 for lysine (AGG), valine (GTC) and isoleucine (ATT) in R-C were replaced by codons for methionine (ATG), isoleucine (ATT) and methionine (ATG) in Rh-131, respectively. No inhibitory effect of pencycuron on the tubulin assembly was observed. It suggests that pencycuron may have no direct inhibitory effects on the assembly of tubulin at least in vitro.

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