• Annals of Clinical Microbiology
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• v.18 no.2
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• pp.37-43
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• 2015

Background: Tuberculosis is globally the most important cause of death from single pathogen. Rapid and accurate identification of mycobacteria is essential for the control of tuberculosis. We evaluated a fluorescence in situ hybridization (FISH) method using peptide nucleic acid (PNA) probes for the differentiation of Mycobacterium tuberculosis complex (MTB) and nontuberculous mycobacteria (NTM) in direct smears of sputum specimens. Methods: The cross-reactivity of MTB- and NTM-specific PNA probes was examined with reference strains of M. tuberculosis ATCC 13950, Mycobacterium kansasii ATCC 12479, Mycobacterium fortuitum ATCC 6841, several clinical isolates of mycobacteria (Mycobacterium abscessus, Mycobacterium avium, Mycobacterium intracellulare, Mycobacterium gordonae and Mycobacterium chelonae), and 11 frequently isolated respiratory bacterial species other than mycobacteria. A series of 128 sputa (89 MTB culture positive, 29 NTM culture positive, and 10 under treatment culture negative) with grades of trace to 4+ were used to evaluate the performance of the method. Results: The MTB- and NTM-specific PNA probes showed specific reactions with the reference strains of MTB and M. kansasii and clinical isolates of mycobacteria except M. fortuitum ATCC 6841, and no cross-reactivity with other tested bacteria. The PNA probe-based FISH assay for detection of MTB had a sensitivity and specificity of 100%, respectively. The sensitivity and specificity of the NTM-specific PNA probe was 100%. The smear grades of the PNA FISH test were same as with those of the fluorescence AFB stain in 2+ or higher grade. Conclusion: Detection and differentiation based on PNA FISH is sensitive and accurate for detecting mycobacteria and for differentiating MTB from NTM in clinical sputum smears.

• Applied Biological Chemistry
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• v.45 no.2
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• pp.53-58
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• 2002

Quinacrine, a pH-sensitive fluorescence probe, which exists either as an unprotonated fluorescence form or a protonated noufluorescence form, can be used to measure the proton transport activity of $H^+-ATPase$. Quinacrine was used to determine the optimal conditions for measuring the activity of microsomal $H^+-ATPase$ prepared from the roots of tomato plants. The amount of quinacrine fluorescence quenching obtained at $0.43{\mu}g/{\mu}l$ of microsomal protein concentration was 25-26%, which shows that the enzyme activity of 100 nmol/min decreases 10% of quinacrine fluorescence. Maximal fluorescence quenching was obtained at pH 7.0-7.2 and 2 mM $Mg^{2+}$ Because the activity of microsomal $H^+-ATPase$ is also maximal at these conditions, the quinacrine fluorescence well represents the activity of $H^+-ATPase$. Vanadate and $NO_3-$, specific inhibitors of plasma and vacuolar $H^+-ATPases$, respectively, were successfully applied to inhibit the quinacrine fluorescence quenching mediated by the corresponding $H^+-ATPases$. These results imply that quinacrine is a useful tool for measuring the proton transport activities of microsomes obtained from the root tissue of tomato plants.

• The Korean Journal of Pharmacology
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• v.31 no.3
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• pp.271-278
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• 1995

The relative distribution ratio of barbiturates between hyarocarbon interior and surface region of outer monolayer of synaptosomal plasma membrane vesicles (RSPMV) isolated from rat whole brain was determined by employing the fluorescent probe technique. The two fluorescent probes N- octadecylnaphthyl-2-amine-6-sulfonic acid (ONS) and 12-(9-anthroyloxy) stearic acid (AS) were utilized as probes for hydrocarbon interior and surface of outer monolayer of RSPMV. respectively. The Stern-Volmer equation for fluorescent quenching was modified to calculate the relative distribution ratio. The analysis of preferential quenching of these probes by barbiturates indicates that pentobarbital, hexobarbital, amobarbital and phenobarbital are predominantly distributed on the surface region. whereas thiopental sodium has an accessibility to the hydrocarbon interior of the outer monolayer of the RSPMV. From these results, it is strongly suggested that the more effective penetration into the hydrocarbon interior of the outer monolayer of the membrane lipid bilayer could result in higher general anesthetic activity.

• Toxicological Research
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• v.34 no.1
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• pp.1-6
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• 2018

The purpose of this study was to detect cell death in the liver of mice treated with thioacetamide (TAA) using fluorescence bioimaging and compare this outcome with that using conventional histopathological examination. At 6 weeks of age, 24 mice were randomly divided into three groups: group 1 (G1), control group; group 2 (G2), fluorescence probe control group; group 3 (G3), TAA-treated group. G3 mice were treated with TAA. Twenty-two hours after TAA treatment, G2 and G3 mice were treated with Annexin-Vivo 750. Fluorescence in vivo bioimaging was performed by fluorescence molecular tomography at two hours after Annexin-Vivo 750 treatment, and fluorescence ex vivo bioimaging of the liver was performed. Liver damage was validated by histopathological examination. In vivo bioimaging showed that the fluorescence intensity was increased in the right upper part of G3 mice compared with that in G2 mice, whereas G1 mice showed no signal. Additionally ex vivo bioimaging showed that the fluorescence intensity was significantly increased in the livers of G3 mice compared with those in G1 or G2 mice (p < 0.05). Histopathological examination of the liver showed no cell death in G1 and G2 mice. However, in G3 mice, there was destruction of hepatocytes and increased cell death. Terminal deoxynucleotidyl transferase dUTP nick end labeling staining confirmed many cell death features in the liver of G3 mice, whereas no pathological findings were observed in the liver of G1 and G2 mice. Taken together, fluorescence bioimaging in this study showed the detection of cell death and made it possible to quantify the level of cell death in male mice. The outcome was correlated with conventional biomedical examination. As it was difficult to differentiate histological location by fluorescent bioimaging, it is necessary to develop specific fluorescent dyes for monitoring hepatic disease progression and to exploit new bioimaging techniques without dye-labeling.

• Genomics & Informatics
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• v.5 no.3
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• pp.113-117
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• 2007

The development of microarray technology has facilitated the understanding of gene expression profiles. Despite its convenience, the cause of dye-bias that confounds data interpretation in dual-color DNA microarray experiments is not well known. In order to economize time and money, it is necessary to identify the cause of dye bias, since designing dye-swaps to reduce the dye-specific bias tends to be very expensive. Hence, we sought to determine the reliable cause of systematic dye bias after treating murine macrophage RAW 264.7 cells with 2-keto-3-deoxyoctonate (KDO), interferon-beta $(IFN-{\beta})$, and 8-bromoadenosine (8-BR). To find the cause of systematic dye bias from the point of view of fluorescence quenching, we examined the correlation between systematic dye bias and the proportion of each nucleotide in mRNA and oligonucleotide probe sequence. Cy3-dye bias was highly correlated with the proportion of adenines. Our results support the fact that systematic dye bias is affected by fluorescence quenching of each feature. In addition, we also found that the strength of fluorescence quenching is based on not only dye-dye interactions but also dye-nucleotide interactions as well.

• Korean Journal of Optics and Photonics
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• v.8 no.4
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• pp.353-355
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• 1997

A single molecule detection system, which consists of confocal fluorescence microscope and single photon counter, has been used to observe the dye concentration dependence of photon counting rate. With increasing concentration, a saturation effect of counting is observed and demonstrated on the basis of the dead time of photon detector. The equations presented here show the relations between the counting rate and some parameters such as probe volume, quantum efficiency of detector, and fluorescence photon number entered onto detector. The signal-to-noise ratio is also discussed briefly.

• Proceedings of the KSAR Conference
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• 2004.06a
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• pp.287-287
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• 2004

Sexing from bovine embryos which were fertilized in vitro implicate a possibility of production of the sex controlled cattle. This study was carried out to investigate the possibility of determining of embryo sex by fluorescence in situ hybridization (FISH) technique. FISH was achieved in in vitro fertilized bovine embryos using a bovine Y-specific DNA probe which constructed from the btDYZ-1 sequences. (omitted)

• Transactions of the Korean Society of Mechanical Engineers A
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• v.31 no.4
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• pp.483-489
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• 2007

We have investigated quantitative measurement techniques of the degree of dispersion of single-walled carbon nanotubes (SWNTs). SWNTs were suspended in aqueous media using a sodium dodecyl sulfate (SDS) surfactant. SWNTs with different dispersion states were prepared by controlling the intensity and time of sonication and centrifugation. The laser spectroscopic techniques were employed to characterize the dispersion state; i.e., raman fluorescence and absorption spectroscopic techniques. Raman spectroscopy has been used to probe the dispersion and aggregation state of SWNTs in solution. Individually suspended SWNTs show increased fluorescence peaks and decreased roping peaks at a raman shift 267 $cm^{-1}$ compared with the samples containing bundles of SWNTs. The ultraviolet-visible-near infrared (UV-vis-NIR) absorption spectrum of decanted supernatant samples show sharp van Hove singularity peaks

• Bulletin of the Korean Chemical Society
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• v.19 no.10
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• pp.1054-1059
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• 1998

Fluorescence and dynamic light scattering measurements were applied to the study of formation and structure of aggregated colloidal particles in modified poly(ethylene-co-methylacrylate) ionomers in aqueous solution. Both 8-anillino-l-naphthalene-sulfonic acid (ANS) and pyrene were used as fluorescence probe to obtain the information on the structure of particle surface and inside, respectively. Three different ionomers used in this study started to aggregate at very dilute concentration, 3-8 x 10-6 g/mL. In this study, we demonstrate that the polyethylene ionomers can form stable nanoparticles. The hydrophobic core made of the polyethylene backbone chains is stabilized by the ionic groups on the particle surface. Such a formed stable nanoparticles have a relatively narrow size distribution with an average radius in the range of 27-48 nm, depending on the kind of ionic groups. Once the stable particles are formed, the particle size distributions were nearly constant. This study shows another way to prepare surfactant-free polyethylene nanoparticles.

• Journal of Powder Materials
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• v.29 no.6
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• pp.453-458
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• 2022

We successfully synthesize water-dispersible CTAB-capped CdSe@ZnS quantum dots with the crystal size of the CdSe quantum dots controlled from green to orange colors. The quenching effect of Fe(DTC)₃ is very efficient to turn off the emission light of quantum dots at four molar ratios of the CdSe quantum dots, that is, the effective covering the surface of quantum dots with Fe(DTC)₃. However, the reaction with Fe(DTC)₃ for more than 24 h is required to completely realize the quenching effect. The highly quenched quantum dots efficiently detect nitric oxide at nano-molar concentration of 110nM of NO with 34% of recovery of emission light intensity. We suggest that Fe(DTC)₃-hybridized CdSe@ZnS quantum dots are an excellent fluorescence resonance energy transfer probe for the detection of nitric oxide in biological systems.