• Title/Summary/Keyword: bio assay

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Quantitative Assay of Bioemulsifier by Turbidometric Method

  • Jeong, Yong-Leen;Park, Oh-Jin;Yoon, Byung-Dae;Yang, Ji-Won
    • Journal of Microbiology and Biotechnology
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    • v.7 no.3
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    • pp.209-211
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    • 1997
  • A quantitative method for assaying bioemulsifiers in culture broth was developed and applied to cultivation of Pseudomonas aeruginosa YPJ80. SED(Standard Emulsification Dilution), an indirect measure of bioemulsifier concentration, was proposed. Production of bioemulsifier and rhamnolipid reached their maximum simultaneously. However, the bioemulsifier/rhamnolipid ratio decreased with cultivation time. This indicates the presence of another bioemulsifier other than rhamnolipid. The bioemulsifier seems to be protein-like activator which showed emulsification activity in addition to rhamnolipid.

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Fabrication of channel-integrated optoelectrofluidic device using stamp-to-stick bonding and microtransfer methods (Stamp-to-Stick Bonding 및 Microtransfer Molding 방법을 이용한 미세유체 채널이 집적된 광전기유체소자의 제작)

  • Hwang, Hyun-Goo;Lee, Do-Hyun;Park, Je-Kyun
    • Journal of Sensor Science and Technology
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    • v.18 no.2
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    • pp.154-159
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    • 2009
  • This paper describes two methods - stamp-to-stick bonding and microtransfer molding - to integrate microfluidic channel into an optoelectrofluidic device for in-channel microparticle manipulation. We have demonstrated the optoelectronic microparticle manipulation in the channel-integrated optoelectrofluidic device using a liquid crystal display. As injecting a liquid sample containing $15{\mu}m$-diameter polystyrene particles into the fabricated channel, trapping and transport of individual microparticles have been successfully demonstrated. This channel-integrated optoelectrofluidic device may be useful for several in-channel applications based on the optoelectrofluidics such as optoelectronic flow control, droplet-based protein assay and bead-based immunoassay.

Immunomodulatory effects of fermented Platycodon grandiflorum extract through NF-κB signaling in RAW 264.7 cells

  • Park, Eun-Jung;Lee, Hae-Jeung
    • Nutrition Research and Practice
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    • v.14 no.5
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    • pp.453-462
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    • 2020
  • BACKGROUND/OBJECTIVES: Platycodon grandiflorum (PG), an oriental herbal medicine, has been known to improve liver function, and has both anti-inflammatory and antimicrobial properties. However, little is known about the immune-enhancing effects of PG and its mechanism. In this study, we aimed to investigate whether fermented PG extract (FPGE), which has increased platycodin D content, activates the immune response in a murine macrophage cell line, RAW 264.7. MATERIALS/METHODS: Cell viability was determined by Cell Counting Kit-8 assay and the nitric oxide (NO) levels were measured using Griess reagent. Cytokine messenger RNA levels of were monitored by quantitative reverse transcription polymerase chain reaction. To investigate the molecular mechanisms underlying immunomodulatory actions of FPGE in RAW 264.7 cells, we have conducted luciferase reporter gene assay and western blotting. RESULTS: We found that FPGE treatment induced macrophage cell proliferation in a dose-dependent manner. FPGE also modulated the expression of NO and pro-inflammatory cytokines, such as tumor necrosis factor-α, interleukin (IL)-1β, and IL-6. The activation and phosphorylation levels of nuclear factor kappa B (NF-κB) were increased by FPGE treatment. Moreover, 5-aminoimidazole-4-carboxamide ribonucleotide, an activator of AMP-activated kinase (AMPK), significantly reduced both lipopolysaccharides- and FPGE-induced NF-κB reporter gene activity. CONCLUSIONS: Taken together, our findings suggest that FPGE may be a novel immune-enhancing agent acting via AMPK-NF-κB signaling pathway.

Optimization of Fabrication Conditions for Immunosensor Strip to Detect Escherichia coli O157 : H7 (Escherichia coli O157 : H7 탐지용 면역센서스트립 제작 조건 최적화)

  • Park, So-Jung;Kim, Young-Kee
    • KSBB Journal
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    • v.24 no.3
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    • pp.253-258
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    • 2009
  • In this study, the optimization of fabrication conditions was accomplished to make immuno-strip biosensor by the combination of enzyme linked immunosorbent assay (ELISA) and immuno-chromatographic strip techniques for the detection of Escherichia coli O157 : H7. Optimal fabrication conditions of capture antibody concentration, detection antibody concentration, and additive composition of running buffer solution were determined. Optimal concentration was determined as 1.0 mg/mL for both of capture antibody and detection antibody. A composition of 0.5% Tween20 and 3% BSA were selected as optimal additive for buffer solution to prevent non-specific binding.

Enhanced Biodegradation of Lindane Using Oil-in-Water Bio-Microemulsion Stabilized by Biosurfactant Produced by a New Yeast Strain, Pseudozyma VITJzN01

  • Abdul Salam, Jaseetha;Das, Nilanjana
    • Journal of Microbiology and Biotechnology
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    • v.23 no.11
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    • pp.1598-1609
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    • 2013
  • Organochlorine pesticide residues continue to remain as a major environmental threat worldwide. Lindane is an organochlorine pesticide widely used as an acaricide in medicine and agriculture. In the present study, a new lindane-degrading yeast strain, Pseudozyma VITJzN01, was identified as a copious producer of glycolipid biosurfactant. The glycolipid structure and type were elucidated by FTIR, NMR spectroscopy, and GC-MS analysis. The surface activity and stability of the glycolipid was analyzed. The glycolipids, characterized as mannosylerythritol lipids (MELs), exhibited excellent surface active properties and the surface tension of water was reduced to 29 mN/m. The glycolipid was stable over a wide range of pH, temperature, and salinity, showing a very low CMC of 25 mg/l. Bio-microemulsion of olive oil-in-water (O/W) was prepared using the purified biosurfactant without addition of any synthetic cosurfactants, for lindane solubilization and enhanced degradation assay in liquid and soil slurry. The O/W bio-microemulsions enhanced the solubility of lindane up to 40-folds. Degradation of lindane (700 mg/l) by VITJzN01 in liquid medium amended with bio-microemulsions was found to be enhanced by 36% in 2 days, compared with degradation in 12 days in the absence of bio-microemulsions. Lindane-spiked soil slurry incubated with bio-microemulsions also showed 20-40% enhanced degradation compared with the treatment with glycolipids or yeast alone. This is the first report on lindane degradation by Pseudozyma sp., and application of bio-microemulsions for enhanced lindane degradation. MEL-stabilized bio-microemulsions can serve as a potential tool for enhanced remediation of diverse lindane-contaminated environments.

Development of Enzyme-Linked Immunosorbent Assay for the Detection of Fumonisins (효소면역측정법을 이용한 Fumonisin의 검출법 개발)

  • Shon, Dong-Hwa;Hahn, Seong-Min;Lim, Sun-Hee;Lee, Yin-Won;Cho, Sun-Hee;Kang, Shin-Young;Lee, Kyung-Ae
    • Microbiology and Biotechnology Letters
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    • v.24 no.1
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    • pp.119-125
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    • 1996
  • In order to develop enzyme-linked immunosorbent assay (ELISA) for fumonisins, production of specific antibodies, establishment of ELISA conditions, and quantitation of the toxin from spiked corns by ELISA were performed. Fumonisin $B_1(FB_1)$ conjugated to cholera toxin (CT) with or without Freund's adjuvant was subcutaneously injected into 2 groups of rabbits. When the titer of the antisera produced by each rabbit was tested, higher titer was observed in case of the immunization with the adjuvant. By use of the antiserum showing the highest titer (1:16,000) and its purified antibodies, competitive indirect and direct ELISA's (ciELISA and cdELISA) were established, respectively. When the cross-reactivity of the antibody against fumonisin analogs was investigated by the ciELISA, it was very low against $B_3$ (2%) but high against fumonisin $B_2$ (179%). The sensitivity of the ELISAs was also very high, because the detection limit for $FB_1$ was 0.03 ppb in ciELISA and 0.3 ppb in cdELISA. When the ELISA's were applied to the spiked corns after extraction with 75% methanol, the assay recovery of $FB_1$ was too unstable to assay. However, when cleanup by strong anion exchange (SAX) cartridge was introduced to remove interfering materials, the mean ELISA recovery of $FB_1$ from corns spiked to 3~10 ppm was found to be 34.0% and stable (mean of CV, 8.2%).

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The Anti-oxidative and Anti-inflammatory Activities of Malus melliana Ethanol Extract (Malus melliana 에탄올 추출물의 항산화 및 항염증 활성)

  • Lee, Su Hyeon;Jin, Kyong-Suk;Kim, Byung Woo;Kwon, Hyun Ju
    • Journal of Life Science
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    • v.27 no.7
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    • pp.783-789
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    • 2017
  • Malus melliana (Hand.-Mazz.) Rehder (M. melliana) is a Chinese plant that belongs to the Rosaceae family. There have been no previous reports regarding its bioactivity. In this study, the anti-oxidative and anti-inflammatory activities of M. melliana ethanol extract (MMEE) were evaluated using a 1,1-diphenyl-2-picryl-hydrazyl (DPPH) radical scavenging activity assay, reactive oxygen species (ROS) scavenging activity assay, nitric oxide (NO) inhibitory activity assay, and the analysis of related protein expressions through Western blot hybridization. MMEE showed potent scavenging activity against DPPH, similar to ascorbic acid, a well-known anti-oxidative agent, which was used as a positive control. MMEE also inhibited hydrogen peroxide-induced ROS in RAW 264.7 cells. Moreover, MMEE induced the expression of an anti-oxidative enzyme, heme oxygenase 1, and its upstream transcription factor, nuclear factor E2-related factor-2, in a dose-dependent manner. On the other hand, MMEE was associated with a reduction in NO production, which was induced by the lipopolysaccharide treatment of RAW 264.7 cells. The expression of inducible nitric oxide synthase, which is the upstream regulator of NO production, was also inhibited. Taken together, these results suggest that MMEE has anti-oxidative and anti-inflammatory properties, thus appearing to be a potential anti-oxidant and anti-inflammatory agent. The further identification of active compounds that confer the biological activities of MMEE may be necessary.

Fluorescence Assay for High Efficient Mass Screening of the Herbicides Inducing Rapid Membrane Peroxidation (막과산화를 신속히 유발하는 제초제의 고효율 대량스크리닝을 위한 형광검정법)

  • Kim, Jin-Seog;Kwon, Ok Kyung
    • Weed & Turfgrass Science
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    • v.4 no.4
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    • pp.308-314
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    • 2015
  • This study was conducted to establish a fluorescence assay system for high efficient mass screening of the herbicides causing rapid membrane peroxidation, based on the fact that peroxide in cellular leakage could be fluorometrically determined through the fuorescent compounds formed after reacting with homovanillic acid (HVA) and peroxidase (HRP). The assay procesure established in this study was as follows. Only single disc (4 mm diameter) excised from cucumber cotyledon is placed on the well containing test solution ($200{\mu}L$) with 96-well microplate. The plate is shaking-incubated for 8 h under light condition. Then after removing the cucumber disc, HVA and HRP are supplied in the medium buffer and incubated for 5 min at room temperature. Fluorescence values are determined at Ex 320 nm/Ex 425 nm. The higher fluorescence values are obtained in the treatment of chemical having higher herbicidal activity. Using this assay with 96-well microplates, a large number of herbicides inducing rapid membrane peroxidation seemed to be screened more efficiently than spectrophotometric microtiter assay reported previously.

Inhibition of Helicobacter pylori Adhesion by Acidic Polysaccharide Isolated from Artemisia capillaris

  • Woo, Jeung-S.;Ha, Byung-H.;Kim, Tae-G.;Lim, Yoon-Gho;Kim, Kyung-H.
    • Journal of Microbiology and Biotechnology
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    • v.13 no.6
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    • pp.853-858
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    • 2003
  • Helicobacter pylori specifically adhere to host cells through a number of putative receptors and ligands, mainly based on carbohydrate-protein interactions. Polysaccharide fractions isolated from the leaves of Artemisia capillaris showed different inhibitory activities against H. pylori adhesion by using hemagglutination assay. Among these fractions, an acidic polysaccharide fraction FlA showed highly effective inhibitory activity, and its minimum inhibition concentration was 0.63 mg/ml. The inhibition results by the hemagglutination assay were consistent with those obtained by the enzymelinked glycosorbent assay, which was developed by the conjugation of horseradish peroxidase with fetuin, a sialic acid-containing glycoprotein which was specific to H. pylori adhesion. FlA contained the highest carbohydrate content among polysaccharide fractions, and no protein was detectable when further purified by gel filtration FPLC. Sugar composition analysis using GC revealed the highest amount of galacturonic acid among sugars, which suggests that FlA contains essentially acidic polysaccharides. Our data suggest that acidic polysaccharides may play an important role in the inhibition of H. pylori adhesion to host cells.

Characteristics of Horizontal Community Distribution and Nutrient Limitation on Growth Rate of Phytoplankton during a Winter in Gwangyang Bay, Korea (동계 광양만에서 식물플랑크톤 군집구조의 수평적 분포특성과 성장에 미치는 영양염 제한 특성)

  • Baek, Seung-Ho;Kim, Dong-Sun;Hyun, Bong-Gil;Choi, Hyun-Woo;Kim, Young-Ok
    • Ocean and Polar Research
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    • v.33 no.2
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    • pp.99-111
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    • 2011
  • To estimate the effects of limitation nutrients for phytoplankton growth and its influences on short-term variations of a winter phytoplankton community structure, we investigated the abiotic and biotic factors of surface and bottom waters at 20 stations of inner and offshore areas from 6 to 7 February in Gwangyang Bay, Korea. Also, several algal bio-assay studies were conducted to identify any additional nutrient effects on phytoplankton assemblage using surface water for the assay. The dominant species in the bay was diatom Skeletonema costatum, which occupied more than 70% of total species in most stations (St.1-16) of the inner bay. According to a cluster and multidimensional scaling (MDS) analysis based on phytoplankton community data from each station, the bay was divided into three groups. The first group included stations from the south-western parts of Myodo lsland, which can be characterized as a semien-closed eutrophic area with high phytoplankton abundance. The second group included most stations from the north-eastern part of Myodo lsland, influenced indirectly by surface water currents from offshore of the bay. The standing phytoplankton crops were lower than those of the first group. The other cluster was restricted to samples collected from offshore of the bay. In the bay, silicon (Si) and phosphorus (P) were not a major limiting factor for phytoplankton production. However, since the DIN: DIP and DSi: DIN ratios clearly demonstrated that there were potential stoichiometric N limitations, nitrogen (N) was considered as a limiting factor. Based on the algal bio-assay, in vivo fluorescence values in N (+) added experiments were higher compared to control and P added experiments. Our results suggested that nitrogen may act as one of the most important factors in controlling primary production during winter in Gwangyang Bay.