• Title/Summary/Keyword: Dye screening

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Development of a Redox Dye-Based Rapid Colorimetric Assay for the Quantitation of Viability/Mortality of Pine Wilt Nematode

  • Han, Kyeongmin;Lee, Jaejoon;Shanmugam, Gnanendra;Lee, Sun Keun;Jeon, Junhyun
    • Journal of Microbiology and Biotechnology
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    • v.29 no.7
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    • pp.1117-1123
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    • 2019
  • Control of pine wilt disease, which is caused by pine wilt nematode Bursaphelenchus xylophilus, is heavily dependent on the use of chemicals such as abamectin. Although such chemicals are highly effective, demands for alternatives that are derived preferentially from natural sources, are increasing out of environmental concerns. One of the challenges to discovery of alternative control agents is lack of fast and efficient screening method that can be used in a high-throughput manner. Here we described the development of colorimetric assay for the rapid and accurate screening of candidate nematicidal compounds/biologics targeting B. xylophilus. Contrary to the conventional method, which relies on laborious visual inspection and counting of nematode population under microscope, our method utilizes a redox dye that changes its color in response to metabolic activity of nematode population in a given sample. In this work, we optimized parameters of our colorimetric assay including number of nematodes and amount of redox dye, and tested applicability of our assay for screening of chemicals and biologics. We demonstrated that our colorimetric assay can be applied to rapid and accurate quantification of nematode viability/mortality in a nematode population treated with candidate chemicals/biologics. Application of our method would facilitate high-throughput endeavors aiming at finding environment-friendly control agents for deadly disease of pine trees.

Coloration of Poly(lactic acid) with Disperse Dyes. 1. Comparison to Poly(ethylene terephthalate) of Dyeability, Shade and Fastness

  • Choi Jae-Hong;Seo Woon-Young
    • Fibers and Polymers
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    • v.7 no.3
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    • pp.270-275
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    • 2006
  • The dyeability of poly(lactic acid) [PLA] with a range of commercial disperse dye was examined and compared to that of poly(ethylene terephthalate) [PET] in addition to the colour and fastness of the resultant dyeings. A screening exercise in which twenty dyes of differing energy types and chemical classes were applied to PLA revealed a substantial variation between the dyes in terms of dye uptake (12-88 % at 4 % o.w.f.). Nine dyes exhausted above 70 % and were selected for further study, which involved comparison of shade and fastness of PLA dyeings with those of the corresponding PET dyeings. Differences in shade depended on hue while wet fastness of each of the PLA dyeings was either the same or 0.5-1.0 point lower than its PET counterpart. In all but one case, dye photostability in PLA was found to be very similar to that in PET. Dye build-up profiles on PLA were also investigated and from these results mixtures of compatible dyes identified.

Simple Analysis for Interaction between Nanoparticles and Dye-Containing Vesicles as a Biomimetic Cell-Membrane

  • Shin, Sohyang;Umh, Ha Nee;Kim, Younghun
    • Bulletin of the Korean Chemical Society
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    • v.34 no.1
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    • pp.231-236
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    • 2013
  • Some cytotoxicity studies for the interpretation of the interaction between nanoparticles and cells are non-mechanistic and time-consuming. Therefore, non-biological screening methods, which are faster and simpler than in-vivo and in-vitro methods, are required as alternatives to current cytotoxicity tests. Here, we proposed a simple screening method for the analysis of the interaction between several AgNPs (bare-, citrate-, and polyvinylpyrrolidone-coating) and dye-containing vesicles acting as a biomimetic cell-membrane. The interaction between AgNPs and vesicles could be evaluated readily by UV-vis spectra. Absorbance deviation in UV-vis spectra revealed a large attraction between neighboring particles and vesicles. This was confirmed by (Derjagin, Landau, Verwey, and Overbeek) theory and DMF (dark-field microscopy) analysis. This proposed method might be useful for analyzing the cytotoxicity of nanoparticles with cell-membranes instead of in vitro or in vivo cytotoxicity tests.

Screening of Biodegradable Function of Indigenous Ligno-degrading Mushroom Using Dyes

  • Jang, Kab-Yeul;Cho, Soo-Muk;Seok, Soon-Ja;Kong, Won-Sik;Kim, Gyu-Hyun;Sung, Jae-Mo
    • Mycobiology
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    • v.37 no.1
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    • pp.53-61
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    • 2009
  • The process of biodegradation in lingo-cellulosic materials is critically relevant to biospheric carbon. The study of this natural process has largely involved laboratory investigations, focused primarily on the biodegradation and recycling of agricultural by-products, generally using basidiomycetes species. In order to collect super white rot fungi and evaluate its ability to degrade lingo-cellulosic material, 35 fungal strains, collected from forests, humus soil, livestock manure, and dead trees, were screened for enzyme activities and their potential to decolorize the commercially used Poly-R 478 dye. In the laccase enzymatic analysis chemical test, 33 white rot fungi and 2 brown rot fungi were identified. The degradation ability of polycyclic aromatic hydrocarbons (PAHs) according to the utilized environmental conditions was higher in the mushrooms grown in dead trees and fallen leaves than in the mushrooms grown in humus soil and livestock manure. Using Poly-R 478 dye to assess the PAH-degradation activity of the identified strains, four strains, including Agrocybe pediades, were selected. The activities of laccase, MnP, and Lip of the four strains with PAH-degrading ability were highest in Pleurotus incarnates. 87 fungal strains, collected from forests, humus soil, livestock manure, and dead trees, were screened for enzyme activities and their potential to decolorize the commercially used Poly-R 478 dye on solid media. Using Poly-R 478 dye to assess the PAHdegrading activity of the identified strains, it was determined that MKACC 51632 and 52492 strains evidenced superior activity in static and shaken liquid cultures. Subsequent screening on plates containing the polymeric dye poly R-478, the decolorization of which is correlated with lignin degradation, resulted in the selection of a strain of Coriolus versicolor, MKACC52492, for further study, primarily due to its rapid growth rate and profound ability to decolorize poly R-478 on solid media. Considering our findings using Poly-R 478 dye to evaluate the PAH-degrading activity of the identified strains, Coriolus versicolor, MKACC 52492 was selected as a favorable strain. Coriolus versicolor, which was collected from Mt. Yeogi in Suwon, was studied for the production of the lignin-modifying enzymes laccase, manganese-dependent peroxidase (MnP), and lignin peroxidase (LiP).

Rapid Screening Method of Peroxidase by Colorimetric Assay and Screening of 2, 4-DCP Degradable Strains (발색법에 의한 Peroxidase의 신속한 스크리닝법과 2, 4-DCP 분해균주의 스크리닝)

  • Ryu, Kang;Lee, Eun-Kyu
    • KSBB Journal
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    • v.23 no.6
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    • pp.484-488
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    • 2008
  • Chlorinated phenols are widely used by the chemical industry as intermediate products in synthesis and previously were frequently applied to various industry fields. Peroxidases catalyze the peroxide-dependent oxidation of a range of inorganic and organic compounds. Peroxidase was shown to mineralize a variety of recalcitrant aromatic compounds and to oxidize a number of polycyclic aromatic and phenolic compounds. Among monomeric phenolic and nonphenolic compounds, peroxidase is known to oxidize its compounds. In this study, a colorimetric assay was developed to quantitatively evaluate the peroxidase activity for rapid screening. Color products of different intensity were developed proportionally to the peroxidase activity on agar plate and 96-well plate. This method correlates well with the RP-HPLC result. Using this screening method, 12 colonies of strain was screened which survived at high concentration of 2,4-DCP (1000 ppm) and with peroxidase activity for the $7^{th}$ round screening step on agar plate. These strains were utilized 2,4-DCP as a sole carbon source and produced peroxidase. After the screening test, four of the bacteria have significant better effect of COD removal on dye waste-water. COD removal of these was from 44% to 61%, respectively.

Simple Analysis for Interaction between Nanoparticles and Fluorescence Vesicle as a Biomimetic Cell for Toxicological Studies

  • Umh, Ha Nee;Kim, Younghun
    • Bulletin of the Korean Chemical Society
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    • v.33 no.12
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    • pp.3998-4002
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    • 2012
  • With continuing progress of nanotechnologies and various applications of nanoparticles, one needs to develop a quick and fairly standard assessment tool to evaluate cytotoxicity of nanoparticles. However, much cytotoxicity studies on the interpretation of the interaction between nanoparticles and cells are non-mechanistic and time-consuming. Here, we propose a simple screening method for the analysis of the interaction between several AgNPs (5.3 to 64 nm) and fluorescence-dye containing vesicles ($12{\mu}m$) acting as a biomimetic cell-membrane. Fluorescence-dye containing vesicle was prepared using a fluorescence probe (1,6-diphenyl-1,3,5-hexatryene), which was intercalated into the lipid bilayer due to their hydrophobicity. Zeta potential of all materials except for bare-AgNPs (+32.8 mV) was negative (-26 to -54 mV). The morphological change (i.e., rupture and fusion of vesicle, and release of dye) after mixing of the vesicle and AgNPs was observed by fluorescence microscopy, and fluorescence image were different with coating materials and surface charge of x-AgNPs. In the results, we found that the surface charge of nanoparticles is the key factor for vesicle rupture and fusion. This proposed method might be useful for analyzing the cytotoxicity of nanoparticles with cell-membranes instead of in vitro or in vivo cytotoxicity tests.

Development of Photo-sensor for Integrated Lab-On-a-Chip (집적화된 Lab-On-a Chip을 위한 광센서의 제작 및 특성 평가)

  • 김주환;신경식;김용국;김태송;김상식;주병권
    • Journal of the Korean Institute of Electrical and Electronic Material Engineers
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    • v.17 no.4
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    • pp.404-409
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    • 2004
  • We fabricated photo-sensor for fluorescence detection in LOC. LOC is high throughput screening system. Our LOC screens biochemical reaction of protein using the immunoassay, and converts biochemical reaction into electrical signal using LIF(Laser Induced Fluorescence) detection method. Protein is labeled with rhodamine intercalating dye and finger PIN photodiode is used as photo-sensor We measured fluorescence emission of rhodamine dye and analyzed tendency of fluorescence detection, according to photo-sensor size, light intensity, and rhodamine concentration. Detection current was almost linearly proportional to two parameters, intensity and concentration, and was inversely proportional to photo-sensor size. Integrated LOC consists of optical-filter deposited photo-sensor and PDMS microchannel detected 50 (pg/${mu}ell$) rhodamine. For integrated LOC including light source, we used green LED as the light source and measured emitted fluorescence.