• Title/Summary/Keyword: Chemical assay

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Quantitative Immunoassay for Polychlorinated Biphenyl Compounds in Electrical Insulating Oils

  • Kim In Soo
    • Proceedings of the Microbiological Society of Korea Conference
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    • 2000.10a
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    • pp.119-127
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    • 2000
  • The development and performance of a competitive indirect immunoassay for the quantitative measurement of polychlorinated biphenyl compounds in insulating oils is presented. Reagent preparation and the assay characterisation, optimisation and validation steps are described. The dynamic range of the assay for Aroclors 1254 and 1260 in methanol was $50-800 {\mu}g\;ml^{-1}$ with $50\%$ signal inhibition values of 217 and $ 212 {\mu}g\;ml^{-1}$ respectively. Impending legislation in the UK is likely to decree that oils containing $ >50 {\mu}g\;ml^{-1}$ PCB be considered contaminated. Assay sensitivity increased with the degree of PCB chlorination. The assay of structurally related compounds of environmental concern yielded cross-reactivity values of under $0.6\%$. The immunoassay proved reliable for the analysis of transformer oils containing $>70{\mu}g\;ml^{-1}$ PCB, but over-estimated PCB levels in oils containing $<20{\mu}g\;ml^{-1}$ of the analyte with the oils requiring pre-treatment using either solid-phase extraction techniques or washing with KOH-ethanol/sulphuric acid to remove matrix interferents. The analytical performance of the assay was compared against a commercially available semi-quantitative immunoassay kit for PCBs in soil and water.

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The Synthesis of New Leuco Squarylium Dyes

  • Park, Soo-Youl;Jun, Kun;Shin, Seung-Rim;Shin, Jong-Il
    • Textile Coloration and Finishing
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    • v.19 no.5
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    • pp.24-29
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    • 2007
  • It is well known that certain classes of dye are reducible to stable colorless leuco compounds, which can be oxidized in air back to the colored species. We now reported that bis(4-dialkylaminophenyl)squaraine dyes are readily reduced in solution by borohydride to give alkali soluble leuco compound, which exists in the 3-hydroxy 2,4-bis(4-dialkylaminophenyl)cyclobuten-one. New alkylamine leuco compounds were synthesized by the reaction of leuco chloro-squaraine with alkylamine derivatives. The leuco compounds are easily isolated and can be air oxidized back to the squaraine dyes. These dyes have many technical application. e.g. in xerography, solar cell, optical recording material, redox indicators, and enzyme assays.

Assessment of decontamination of gutta-percha cone and the change of surface texture after rapid chemical disinfection (화학소독제 처리 후 가타파차 콘의 멸균 효과 및 표면 성상의 변화 평가)

  • Pang, Nan-Sim;Jung, Il-Young;Yu, Yoon-Jung;Kum, Kee-Yeon
    • Restorative Dentistry and Endodontics
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    • v.31 no.2
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    • pp.133-139
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    • 2006
  • The purposes of this study were firstly to identify the microbial species on gutta-percha (GP) cones exposed at outpatient clinics using polymerase chain reaction, and secondly to evaluate the rapid sterilization effect of two chemical disinfectants at chair side. It also evaluated the alteration of surface texture of GP cones after 5-min soaking into two chemical disinfectants. A total of 100 GP cones from two endodontic departments were randomly selected for microbial detection using PCR assay with universal primer. After inoculation on the sterilized GP cones with the same microorganism identified by PCR assay, they were soaked in two chemical disinfectants: 5% NaOCl and 2% chlorhexidine for 1, 3, 5, and 10 minutes. The sterilization effect was evaluated by turbidity and subculture. The change of surface textures using a scanning electron microscope was also examined after 5 min-soaking in two chemical disinfectants. Results showed that four bacterial species were detected in 17 GP cones, and all the species belonged to the genus Staphylococcus. Two chemical disinfectants were effective in sterilization with just 1 minute soaking. On the SEM picture of NaOCl-soaked GP cone, a cluster of cuboidal crystals was seen on the cone surface. Present data demonstrate that two chemical disinfectants are useful for rapid sterilization of GP cone just before obturation at chair side, while CHX-soaked GP cone has cleaner surface without crystal precipitation than that of NaOCl-treated cone.

Comparison of Digital PCR and Quantitative PCR with Various SARS-CoV-2 Primer-Probe Sets

  • Park, Changwoo;Lee, Jina;Hassan, Zohaib ul;Ku, Keun Bon;Kim, Seong-Jun;Kim, Hong Gi;Park, Edmond Changkyun;Park, Gun-Soo;Park, Daeui;Baek, Seung-Hwa;Park, Dongju;Lee, Jihye;Jeon, Sangeun;Kim, Seungtaek;Lee, Chang-Seop;Yoo, Hee Min;Kim, Seil
    • Journal of Microbiology and Biotechnology
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    • v.31 no.3
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    • pp.358-367
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    • 2021
  • The World Health Organization (WHO) has declared the coronavirus disease 2019 (COVID-19) as an international health emergency. Current diagnostic tests are based on the reverse transcription-quantitative polymerase chain reaction (RT-qPCR) method, which is the gold standard test that involves the amplification of viral RNA. However, the RT-qPCR assay has limitations in terms of sensitivity and quantification. In this study, we tested both qPCR and droplet digital PCR (ddPCR) to detect low amounts of viral RNA. The cycle threshold (CT) of the viral RNA by RT-PCR significantly varied according to the sequences of the primer and probe sets with in vitro transcript (IVT) RNA or viral RNA as templates, whereas the copy number of the viral RNA by ddPCR was effectively quantified with IVT RNA, cultured viral RNA, and RNA from clinical samples. Furthermore, the clinical samples were assayed via both methods, and the sensitivity of the ddPCR was determined to be equal to or more than that of the RT-qPCR. However, the ddPCR assay is more suitable for determining the copy number of reference materials. These findings suggest that the qPCR assay with the ddPCR defined reference materials could be used as a highly sensitive and compatible diagnostic method for viral RNA detection.

Antioxidant Activity and Whitening Efficacy of Makgeolli Fractions (막걸리 분획물의 항산화활성 및 미백효능)

  • Park, Kyung-won;Kwak, Da-hee;Kwon, Hye-Jin
    • Journal of Digital Convergence
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    • v.15 no.10
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    • pp.571-577
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    • 2017
  • This study aimed at evaluating the potential of makgeolli, which is widely consumed as beverage, as a functional cosmetic ingredient, based on analyses on its antioxidant activity and tyrosinase activity inhibitory effect. The sample was extracted by concentrating the suspension obtained after adding MeOH (3 L) to the residue of a commercial makgeolli and then adding EtOAc (3 L) to the concentrate, which was subjected to fractionation. The upper layer of the fractions was used as the final sample. In MTT assay assessments, no cytotoxicity was observed at a concentration range of 10 to $1,000{\mu}g/mL$; the antioxidant activity of the extract showed a concentration-dependent tendency and it had a high activity with an $EC_{50}$ of 7.008 mg/mL. Also, in a tyrosinase activity inhibitory effect assessment, the extract showed an $IC_{50}$ value of 39.22 mg/mL. These results confirmed that this sample has potential as a functional cosmetic ingredient.

Chemical compositions and biological activities of marine invertebrates from the East Sea of South Korea

  • Kim, Geun-Hyeong;Park, Hyeon-Ho;Chandika, Pathum;Ko, Seok-Chun;Jung, Kyung-Mi;Yoon, Sang Chul;Oh, Taeg-Yun;Kim, Young-Mog;Jung, Won-Kyo
    • Fisheries and Aquatic Sciences
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    • v.22 no.6
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    • pp.13.1-13.9
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    • 2019
  • Background: Marine invertebrates are well known as pivotal bioresources with bioactive substances such as anti-inflammatory sterols, antitumor terpenes, and antimicrobial peptides. However, there are few scientific reports on chemical compositions and bioactivities of marine invertebrates from the East Sea of South Korea. Methods: In this study, chemical compositions and biological activities were evaluated on both 70% EtOH and hot water extracts of 5 species of marine invertebrates (Crossaster papposus japonicus, Actinostola carlgreni, Stomphia coccinea, Actinostola sp., and Heliometra glacialis) collected from the East Sea of South Korea. The antioxidant activities were measured by ABTS radical scavenging assay. The cytotoxicity and anti-inflammatory activity were evaluated using MTT and Griess reagents. Moreover, the antibacterial effect was evaluated using paper disc assay and minimum inhibitory concentration (MIC) assay. Results: In the results of antioxidant activities, 70% EtOH extract of A. carlgreni showed the highest activity ($IC_{50}\;0.19{\pm}0.03mg/ml$) compared to other extracts. Moreover, 70% EtOH extract of A. carlgreni could significantly suppress the nitric oxide (NO) production in lipopolysaccharide-induced RAW 264.7. All extracts treated under $400{\mu}g/ml$ have no cytotoxic effects on RAW 264.7 macrophages. In the antibacterial test, both 70% EtOH extracts of C. papposus japonicus and H. glacialis showed a significant antibacterial effect on Staphylococcus aureus. The MIC values were evaluated at 256 and $512{\mu}g/ml$, respectively. Conclusions: These results suggested the bioactive potentials of marine invertebrates from the East Sea of South Korea in pharmaceutical and nutraceutical applications.

Development of a Sensitive Bioassay Method for Quorum Sensing Inhibitor Screening Using a Recombinant Agrobacterium tumefaciens

  • Kim Yeon Hee;Kim Young Hee;Kim Jung Sun;Park Sunghoon
    • Biotechnology and Bioprocess Engineering:BBE
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    • v.10 no.4
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    • pp.322-328
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    • 2005
  • Acylhomoserine lactones (AHLs) are known to be the triggering molecules in the quorum sensing mechanism of many gram-negative bacteria. In order to detect AHL inhibitors that are potential biofilm inhibitors, a convenient and sensitive bioassay was developed based on the $\beta$-galactosidase activity ($\beta$-GAL) of a recombinant Agrobacterium tumefaciens strain. A series of commercially available AHLs were tested for inducing $\beta$-GAL at varying concentrations in agar-plate and liquid cultures of the reporter strain. All AHLs tested exhibited a concentration­dependent induction, and octanoyl homoserine lactone (OHL) showed the highest sensitivity with a detection limit of 0.1 nM in the liquid culture assay. When fimbrolide, a known quorum sensing inhibitor, was added, induction of $\beta$-GAL by OHL was repressed. The repression at a constant OHL concentration was dependent on the fimbrolide concentration with the detection limit below 1 ppm, indicating that this assay is a sensitive method for screening AHL inhibitors.

Biosurfactant Production from Novel Air Isolate NITT6L: Screening, Characterization and Optimization of Media

  • Vanavil, B.;Perumalsamy, M.;Rao, A. Seshagiri
    • Journal of Microbiology and Biotechnology
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    • v.23 no.9
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    • pp.1229-1243
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    • 2013
  • In this paper, an air isolate (NITT6L) has been screened based on hemolytic activity, emulsification activity, drop collapsing test, and oil displacement test, as well as lipase activity. It was found that strain NITT6L was able to reduce the surface tension of the medium from 61.5 to 39.83 mN/m and could form stable emulsions with tested vegetable oils. Morphological, biochemical, 16S rRNA sequencing analyses, and fatty acid methyl ester analysis using gas chromatography confirmed that the air isolate under study was Pseudomonas aeruginosa. Characterization of the biosurfactant using agar double diffusion assay revealed that the biosurfactant was anionic in nature, and CTAB-methylene blue assay and Molisch test revealed its glycolipid nature. The FT-IR spectrum confirmed that the crude biosurfactant was a rhamnolipid. Using unoptimized medium containing sucrose as the carbon source, the isolate was found to produce 0.3 mg/ml of rhamnolipid in batch cultivation (shake flask) at $37^{\circ}C$ and pH 7. Optimization of the medium components was carried out using design of experiments and the yield of rhamnolipid has been enhanced to 4.6 mg/ml in 72 h of fermentation.

Distinct $[^3H]$MK-801 Binding Profiles with the Agonist, Partial Agonist, and Antagonist Acting at the Glycine Binding Site of the N-Methyl-D-Aspartate Receptor

  • Cho, Jung-sook;Park, No-Sang;Kong, Jae-Yang
    • Biomolecules & Therapeutics
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    • v.4 no.2
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    • pp.196-201
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    • 1996
  • The N-methyl-D-aspartate (NMDA) receptor-ion channel complex is activated by the simultaneous presence of L-glutamate and glycine, allowing the binding of MK-801 to the phencyclidine (PCP) site of the receptor. The $[^3H]$MK-801 binding assay system was established for determination of pharmacological functions of test compounds acting at the glycine site of the receptor. The binding in the presence of 0.1 $\mu$M L-glutamate was increased by an agonist (glycine) in a dose-dependent fashion, while decreased by either partial agonist (R-(+)-HA-966) or antagonist (5,7-dichlorokynurenic acid: 5,7-DCKA). To distinguish partial agonism from antagonism, various concentrations of 7-chlorokynurenic acid (7-CKA) were added in the assay to eliminate the interference of the endogenous glycine present in the membrane preparations. The bindings in the presence of L-glutamate (0.1$\muM$) and 7-CKA (1, 5, or 10$\muM$) were increased by R-(+)-HA-966. Being a weak partial agonist, the extent of potentiation was much less than that by the agonist. These binding profiles were clearly distinguishable from those by the antagonist, 5,7-DCKA, which exhibited no intrinsic activity. The binding assays established in the present study are a useful system to classify ligands acting at the glycine site of the NMDA receptor by their pharmacological functions.

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