• Title/Summary/Keyword: Bioassays

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Examination of $\alpha$-terpinene on Primary Eye Irritancy and Skin Sensitization

  • Park, Byeoung-Soo;Choi, Won-Sik;Lee, Sung-Eun
    • Molecular & Cellular Toxicology
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    • v.3 no.1
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    • pp.68-75
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    • 2007
  • [ $\alpha$ ]-Terpinene has been known as a repellent against the mosquito Culex pipiens pallens Coquillett based on a human forearm bioassay. $\alpha$-Terpinene showed significantly greater repellency than a commercial formulation, N, N-diethyl-m-methylbenzamide (deet). In this study, skin and eye sensitivity of $\alpha$-terpinene (2%) was examined with bioassays using white New Zealand rabbits. There were somewhat gross and histological changes observed in these treatments. Eye irritancy assays examined gross changes to cornea, iris and conjuctiva, and histological changes to smear of ocular discharge and eye tissue. Treated rabbits were divided into two cohorts, a saline washed cohort (W) or a non-washed cohort (NW). Opacity of cornea and redness, chemosis and discharge of conjuctiva were observed in both cohorts, but disappeared within 4 and 10 days in W and NW, respectively. Main components of ocular discharges were fibrin, epithelial or epitheloid cells, lymphoid cells, erythrocytes and granulocytes. These abnormal cellular components disappeared within 4 days and 10 days in W and NW, respectively. No permanent histological differences were observed between the two cohorts. However, severe irritation was determined as 57.2 of I.I.O.I value on the first day after treatment. These findings indicate a spray-type solution containing 2% $\alpha$-terpinene may serve as an alternative mosquito repellent and further studies need to reduce the eye irritation with formulation changes.

Phytotoxic Effect of Xanthium occidentale Leaf Extract on Seed Germination and Early Seedling Growth of Alfalfa and Barnyard Grass

  • Chon, Sang-Uk
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.49 no.1
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    • pp.30-35
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    • 2004
  • Compositae plants are known to contain biologically active substances that are allelopathic to agricultural crops as well as weed species. Aqueous extracts from leaves of Xanthium occidentale were assayed against alfalfa (Medicago sativa) to determine their allelopathic effects, and the result showed that the extracts applied onto filter paper significantly inhibited seed germination as well as root growth of alfalfa. Untreated seeds germinated in 60h, but extract concentrations greater than 30g $\textrm{L}^{-1}$ delayed seed germination. The extracts significantly inhibited seed germination of alfalfa, and $\beta$-amylase activity of alfalfa and barley seeds during 24-36 hours after treatment. Aqueous extracts of 40 g $\textrm{L}^{-1}$ from X. occidentale were completely inhibited the hypocotyl and root growth of alfalfa. Aqueous leaf extracts showed the highest inhibitory effect and followed by root and stem extracts. Early seedling growth of both alfalfa and barnyard grass (Echinochloa crus-galli) was significantly reduced by methanol extracts. By means of high-performance liquid chromatography, chlorogenic acid and trans-cinnamic acid were quantified as the highest amounts from water and EtOAc fractions, respectively. BuOH and EtOAc fractions of X. occidentale reduced alfalfa root growth more than did hexane and water fractions. The findings of the bioassays for aqueous or methanol extracts reflected that the inhibitory effect of extract was closely related to the level of responsible allelochemicals found in plant extracts.

Control Efficacy of Fungicides on Cherry Witches' Broom Caused by Taphrina wiesneri (벚나무 빗자루병균(Taphrina wiesneri)에 대한 살균제의 방제효과)

  • Seo, Sang-Tae;Kim, Kyung-Hee;Shin, Chang-Hoon;Lee, Sang-Hyun;Kim, Young-Mal;Park, Jong-Han;Shin, Sang-Chul
    • Research in Plant Disease
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    • v.15 no.1
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    • pp.13-16
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    • 2009
  • In April 2008, cherry (Prunus yedoensis Matsumura) in Jeju exhibited severe disease of branches forming numerous twigs. Five isolates from diseased leaves were identified as Taphrina wiesneri on the basis of biological and genetic characteristics. Twenty one commercial fungicides were tested for control of the disease in vitro. Of the 21 fungicides, triazole compounds (difenoconazole, propiconazole and tebuconazole) showed relatively good antimicrobial activities. Results from the planta bioassays indicated that triazole compounds reduced the witches' broom disease showing control value $25.7{\sim}52.8%$ compared with the nontreated controls.

Enhanced Biological Control of Phytophthora Blight of Pepper by Biosurfactant-Producing Pseudomonas

  • Ozyilmaz, Umit;Benlioglu, Kemal
    • The Plant Pathology Journal
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    • v.29 no.4
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    • pp.418-426
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    • 2013
  • Pseudomonas isolates from different crop plants were screened for in vitro growth inhibition of Phytophthora capsici and production of biosurfactant. Two in vivo experiments were performed to determine the efficacy of selected Pseudomonas strains against Phytophthora blight of pepper by comparing two fungicide treatments [acibenzolar-S-methyl (ASM) and ASM + mefenoxam]. Bacterial isolates were applied by soil drenching ($1{\times}10^9$ cells/ml), ASM ($0.1{\mu}g$ a.i./ml) and ASM + mefenoxam (0.2 mg product/ml) were applied by foliar spraying, and P. capsici inoculum was incorporated into the pot soil three days after treatments. In the first experiment, four Pseudomonas strains resulted in significant reduction from 48.4 to 61.3% in Phytophthora blight severity. In the second experiment, bacterial treatments combining with olive oil (5 mL per plant) significantly enhanced biological control activity, resulting in a reduction of disease level ranging from 56.8 to 81.1%. ASM + mefenoxam was the most effective treatment while ASM alone was less effective in both bioassays. These results indicate that our Pseudomonas fluorescens strains (6L10, 6ba6 and 3ss9) that have biosurfactant-producing abilities are effective against P. capsici on pepper, and enhanced disease suppression could be achieved when they were used in combination with olive oil.

Characterization of Quorum-Sensing Signaling Molecules Produced by Burkholderia cepacia G4

  • Park, Jun-Ho;Hwang, In-Gyu;Kim, Jin-Wan;Lee, Soo-O;Conway, B.;Peter Greenberg, E.;Lee, Kyoung
    • Journal of Microbiology and Biotechnology
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    • v.11 no.5
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    • pp.804-811
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    • 2001
  • In many Gram-negative bacteria, autoinducers, such as N-acyl-L-homoserine lactone(acyl-HSL) and its derivative molecules, mediate the cell-density-dependnet expression of certain operons. The current study identified the autoinducers produced by Burkholderia cepacia G4, a trichloroethylene-degrading lagoon isolate, using TLC bioassays with Agrobacterium tumefaciens NT1(pDCI141E33) and Chromobacterium violaceum CVO26, and a GC-MS analysis. The ${R_f}\;and\;{R_t}$ values and mass spectra were compared with those of synthetic compounds. Based on the analyses, it was confirmed that G4 produces N-hexanoyl (C6)-, N-octanoyl (C8)-, N-decanoyl (C10)-, N-dodecanoyl (C12)-HSL, and an unknown active species. The integration of the GC peak areas exhibited a ratio of C8-HSL:C10-HSL:C12-HSL at 3:17:1 with C6-HSL and C10-HSL production at trace and micromolar levels, respectively, in the culture supernatants. Nutants partially defective in producing acyl-HSLs were also partially defective in the biosynthesis of an antibiotic substance. These results indicate that the autoinducer-dependent gene regulation in G4 is dissimilar to the clinical B. cepacia strains isolated from patients with cystic fibrosis.

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Screening for Herbs Having Inhibitory Effect on Egg Hatching of Strongyloides venezuelensis

  • Kim, Byeong-Soo;Baek, Byeong-Kirl
    • Biomedical Science Letters
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    • v.8 no.4
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    • pp.223-228
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    • 2002
  • In vitro bioassays of aqueous extracts of 25 herbal species were performed by ovicidal activity against Strongvloides venuelensis. 500 mg of feces from Mongorian gerbil infected with S. venuelensis were placed into polyvinyl culture with extracts of herb and anthelmintic drugs in triplicates. Larval activity inside the polyvinyl bag was recorded microscopically at 12 hours, and every 24 hours thereafter for up to 72 hrs incubation. Eggs were not hatched within 72 hrs and degeneration was evident in the presence of extracts. The ovicidal activity characterized by the absence of L$_3$ in fecal cultures inside the polyvinyl bag. 1. Among 25 herbs, 12 herbs had ovicidal effects and caused degeneration of larvae in egg. S. angustifolia, I. helenium, A. korenum, P. tenuifolia, F. virridissima, A. asiatica, S. chinensis, A. tataricus, S. baicalensis, M. rimosa, S. glabra and Z. piperitum were the most ovicidal effect. However, P. thunbergiana, T. chinensis, C. flos, G. uralensis, C. obutusifolia, C. fortunei, A. koreana, P. grandiflorum, P. tenuifolia (Radix), P. mume and A. asphodelodes showed no ovicidal effects in vinyl bag culture. 2. Of 25 hrebs, 4 herbs had ovicidal and wormicidal effects, P. thunbergiana, C. flos, B. Striata and T. chinensis were characterized by degeneration of larvae.

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Estogenic and Dioxin-like Activity of Influent and Effluent of the Industrial Wastewater Treatment Plant

  • Oh, Seung-Min;Kim, Gi-Sur;Kim, Soung-Ho;Kim, Yun-Hee;Chung, Kyu-Hyuck
    • Proceedings of the Korea Society of Environmental Toocicology Conference
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    • 2002.10a
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    • pp.166-166
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    • 2002
  • The response of environmental pollutants can be detected bioanalytically focusing on the source and matrices of concern. Cell culture bioassays are rapid and inexpensive, and thus have great potential for determination of environmental pollution. We have examined the estrogenic and dioxin-like activites of industrial wastewater using E-screen assay and EROD microbioassay. Influent and effluent wastewater were collected from four different industrial wastewater treatment plants, such as cosmetics, paints, textile producing and metal coating plant, and extracted using solid-phase extraction with Oasis@HLB plus cartridge. Pollutants adsorbed to the cartridge were eluted with MTBE. MCF-7 cells were treated with extracts showed various estrogenic potential. The textile wastewater showed strong estrogenic activity and the others showed weak estrogenic activity, No effect was observed in the wastewater from paints producing plant. All extracts showed CYPIA inducing effects, indicating these samples contain dioxin-like chemicals. Bioanalytical results of effluents compared with influents could give us information about the incomplete wastewater treatment and biological potency caused by pollutants. [Supported by a Grant from the Korea Science and Engineering Foundation]

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Flavonols from Houttuynia cordata with Protein Glycation and Aldose Reductase Inhibitory Activity

  • Jang, Dae-Sik;Kim, Jong-Min;Lee, Yun-Mi;Yoo, Jeong-Lim;Kim, Young-Sook;Kim, Joo-Hwan;Kim, Jin-Sook
    • Natural Product Sciences
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    • v.12 no.4
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    • pp.210-213
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    • 2006
  • A 4,5-dioxoaporphine type alkaloid, cepharadione B (1), a phenolic acid, protocatechuic acid (2), and flavonols, quercetin (3), afzelin (4), and quercitrin (5), were isolated from the EtOAc-soluble extract of the whole plants of Houttuynia cordata. All the isolates (1-5) were subjected to in vitro bioassays to evaluate advanced glycation end products (AGEs) formation and rat lens aldose reductase (RLAR) inhibitory activity. The three flavonols 3-5 exhibited a significant inhibitory activity on AGEs formation with $IC_{50}$ values of 66.9, 58.9, and $32.3{\mu}M$, respectively. While the two flavonol rhamnosides 4 and 5 showed a remarkable inhibitory activity against RLAR with $IC_{50}$ values of 0.81 and $0.16{\mu}M$, respectively.

Identification and Characterization of Novel Biocontrol Bacterial Strains

  • Lee, Seung Hwan;Kim, In Seon;Kim, Young Cheol
    • Research in Plant Disease
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    • v.20 no.3
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    • pp.182-188
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    • 2014
  • Because bacterial isolates from only a few genera have been developed commercially as biopesticides, discovery and characterization of novel bacterial strains will be a key to market expansion. Our previous screen using plant bioassays identified 24 novel biocontrol isolates representing 12 different genera. In this study, we characterized the 3 isolates showing the best biocontrol activities. The isolates were Pantoea dispersa WCU35, Proteus myxofaciens WCU244, and Exiguobacterium acetylicum WCU292 based on 16S rRNA sequence analysis. The isolates showed differential production of extracellular enzymes, antimicrobial activity against various fungal or bacterial plant pathogens, and induced systemic resistance activity against tomato gray mold disease caused by Botrytis cinerea. E. acetylicum WCU292 lacked strong in vitro antimicrobial activity against plant pathogens, but induced systemic resistance against tomato gray mold disease. These results confirm that the trait of biological control is found in a wide variety of bacterial genera.

ATP-independent Thermoprotective Activity of Nicotiana tabacum Heat Shock Protein 70 in Escherichia coli

  • Cho, Eun-Kyung;Bae, Song-Ja
    • BMB Reports
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    • v.40 no.1
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    • pp.107-112
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    • 2007
  • To study the functioning of HSP70 in Escherichia coli, we selected NtHSP70-2 (AY372070) from among three genomic clones isolated in Nicotiana tabacum. Recombinant NtHSP70-2, containing a hexahistidine tag at the amino-terminus, was constructed, expressed in E. coli, and purified by $Ni^{2+}$ affinity chromatography and Q Sepharose Fast Flow anion exchange chromatography. The expressed fusion protein, $H_6NtHSP70$-2 (hexahistidine-tagged Nicotiana tabacum heat shock protein 70-2), maintained the stability of E. coli proteins up to 90$^{\circ}C$. Measuring the light scattering of luciferase (luc) revealed that NtHSP70-2 prevents the aggregation of luc without ATP during high-temperature stress. In a functional bioassay (1 h at 50$^{\circ}C$) for recombinant $H_6NtHSP70$-2, E. coli cells overexpressing $H_6NtHSP70$-2 survived about seven times longer than those lacking $H_6NtHSP70$-2. After 2 h at 50$^{\circ}C$, only the E. coli overexpressing $H_6NtHSP70$-2 survived under such conditions. Our NtHSP70-2 bioassays, as well as in vitro studies, strongly suggest that HSP70 confers thermo-tolerance to E. coli.