• Title/Summary/Keyword: fluorescence values

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In vitro Folding of Recombinant Hepatitis B Virus X-Protein Produced in Escherichia coli: Formation of Folding Intermediates

  • Kim, Sun-Ok;Sohn, Mi-Jin;Jeong, Soon-Seog;Shin, Jeh-Hoon;Lee, Young-Ik
    • BMB Reports
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    • v.32 no.6
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    • pp.521-528
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    • 1999
  • The folding of recombinant hepatitis B virus X-protein (rHBx) solubilized from Escherichia coli inclusion bodies was investigated. By sequential dialysis of urea, rHBx was folded into its native structure, which was demonstrated by the efficacy of its transcriptional activation of the adenovirus major late promoter (MLP), fluorescence spectroscopy, and circular dichroism (CD) analysis. The decrease in CD values at 220 nm and a corresponding blue shift of the intrinsic fluorescence emission confirmed the ability of rHBx to refold in lower concentrations of urea, yielding the active protein. Equilibrium and kinetic studies of the refolding of rHBx were carried out by tryptophan fluorescence measurements. From the biphasic nature of the fluorescence curves, the existence of stable intermediate states in the renaturation process was inferred. Reverse phase-high performance liquid chromatography (RP-HPLC) analysis further demonstrated the existence of these intermediates and their apparent compactness.

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Fluorescence Quenching of Bis-msb by Carbon Tetrachloride in Different Solvents

  • Thipperudrappa, J.;Biradar, D.S.;Lagare, M.T.;Hanagodimath, S.M.;Inamdar, S.R.;Kadadevaramath, J.S.
    • Journal of Photoscience
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    • v.11 no.1
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    • pp.11-17
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    • 2004
  • Fluorescence quenching of l,4-bis [2-(2-methylphenyl) ethenyl]-benzene (Bis-MSB) by carbon tetrachloride in five different solvents namely hexane, cyclohexane, toluene, benzene and dioxane has been carried out at room temperature with a view to understand the quenching mechanisms. The Stern-Volmer plot has been found to be non-linear with a positive deviation for all the solvents studied. In order to interpret these results we have invoked the Ground state complex and Sphere of action static quenching models. Using these models various rate parameters have been determined. The magnitudes of these parameters imply that sphere of action static quenching model agrees well with the experimental results. Hence the positive deviation in the Stem-Volmer plots is attributed to the static and dynamic quenching. Further, with the use of Finite Sink approximation model, it was possible to check whether these bimolecular reactions as diffusion limited and to estimate independently distance parameter R' and mutual diffusion coefficient D. Finally an effort has been made to correlate the values of R'and D with the values of the encounter distance R and the mutual diffusion coefficient D determined using the Edwardis empirical relation and Stokes-Einstein relation.

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Symmetric Bis-Azospiropyrans: Synthesis, Characterization and Colorimetric Study

  • Nourmohammadian, Farahnaz;Abdi, Ali Ashtiani
    • Bulletin of the Korean Chemical Society
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    • v.34 no.6
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    • pp.1727-1734
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    • 2013
  • Synthesis and characterization of some novel symmetric bis-azospiropyrans are reported in this study. These bis-azospiropyrans are bifunctional chromophores with two spiropyrans linked by a bis-azo extended aromatic system that produce more color strength (large molar absorption coefficient in mero forms) due to appending two azospiropyran chromophores on one molecule. Comparing to the molar absorption coefficients of the conventional spiropyran chromophores (${\varepsilon}=0.31{\times}10^4\;M^{-1}{\cdot}cm^{-1}$) and mono-azospiropyran chromophores ($1.35{\times}10^4\;M^{-1}{\cdot}cm^{-1}$), the novel synthesized photochromes showed astonishingly increased molar absorption coefficients ($2.3-3.8{\times}10^4\;M^{-1}{\cdot}cm^{-1}$) at the same conditions. Such high molar absorption coefficients confers high sensitivity to light and more color intensity of mero form, that leads to improvement of their light sensitivity and better discrimination of spiro (OFF) form from mero (ON) ones in molecular switches. The structures were deduced from their MS, FT-IR, and $^1H$-NMR spectroscopic data and CHN analysis. All the synthesized photochemically bifunctional compounds revealed fluorescent emission in their colorless form which was faded out after exposing to UV light. Fluorescence quantum yield values of the mero forms were 0.25-0.81 and two high fluorescence quantum yield values (0.60 and 0.81) were found in these series.

An Intercomparison of Atmospheric SO$_{2}$ Measurement Techniques (대기중 이산화황 관측 방법들의 실측 비교 연구)

  • 김경렬;이동수;이강웅
    • Journal of Korean Society for Atmospheric Environment
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    • v.12 no.5
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    • pp.587-591
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    • 1996
  • An intercomparison experiment was implemented to assess the uncertainties and precisions om atmospheric SO$_{2}$ measurement techiques including a pulsed fluorescence (P-F), a diffusion-scrubber /ion chromatography (D-I), and a mist-chamber/ion chromatography (M-I). Each of those three techniques was investigated by researchers at Seoul National University, Yonsei University, and Hankuk University of Foreign Studies, respectively. The concentrations of atmospheric SO$_{2}$ were determined concurrently using three independent measurement techniques at the Seoul National University campus, Korea during Nov.22 to Dec.2, 1995. While the results from the P-F and D-I techinques showed the very close agreements (within +-5%) throughout the experiment period, M-I technique showed systematically smaller values (up to 30%) than the other two techniques. Although sources of larger discrepancy between different techniques were mot identified, the lower SO$_{2}$ values of the M-I method may be related to the errors associated with sample collection effciency, mass flow rate measurements, and standardization of ion chromatography.

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FLUORESCENCE QUENCHING OF BBOT BY ANILINE IN DIFFERENT ORGANIC SOLVENTS

  • Kadadevarmath, J.S.;Giraddi, T.P.;Chikkur, G.C.
    • Journal of Photoscience
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    • v.4 no.3
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    • pp.105-112
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    • 1997
  • The fluorescence quenching of 2, 5-di-(5-tert-butyl-2-benzoxazolyl)-thiophene (BBOT) by aniline in five different solvents namely heptane, hexane, cyclohexane, dioxane and acetonitrile has been carried out at room temperature with a view to understand the quenching mechanisms. The experimental results show positive deviation in the Stern-Volmer plots in all the solvents. In order to interpret these results we have invoked the Ground state complex and sphere of action static quenching models. Using these models various rate parameters have been determined. The magnitudes of these parameters suggest that sphere of action static quenching model agrees well with the experimental results. Hence this positive deviation is attributed to the static and dynamic quenching. Further, with the use of Finite Sink approximation model, it was possible to check these bimolecular reactions as diffusion-limited and to estimate independently distance parameter R$^1$ and mutual diffusion coefficient D. Finally an attempt has been made to correlate the values of R$^1$ and D with the values of the encounter distance R and the mutual diffusion coefficient D determined using the Edward's empirical relation and Stokes-Einstein relation.

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A Study on Spectra of Laser Induced Flourescence in Phantom of N-propyl-N,N-dimethylethanolamine (N-propyl-N,N-dimethylethanolamine의 Phantom에서 Laser Induced Fluorescence의 스펙트라에 관한 연구)

  • Kim, Ki-Jun;Lee, Joo-Ho;Lee, Joo-Youb;Sung, Wan-Mo
    • Journal of the Korean Applied Science and Technology
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    • v.32 no.2
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    • pp.330-338
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    • 2015
  • The influences of fluorescence, scattering, and flocculation in turbid material by light scattering of N-propyl-N,N-dimethylethanolamine, fluorescence agent and absorption agent were interpreted for the scattered fluorescence intensity and wavelength. They have been studied the molecular properties by the spectroscopy of laser induced fluorescence (LIF) and flocculation. The effects of optical properties in scattering media have been found by the optical parameters(${\mu}_s$, ${\mu}_a$, ${\mu}_t$). Flocculation is an important step in many solid-liquid separation processes and is widely used. When two particles approach each other, interactions of several colloid particles can come into play which may have major effect on the flocculation and LIF process. The values of scattering coefficient ${\mu}_s$ are large by means of the increasing scattering of scatterer, The values have been found that the slope decays exponentially as a function of concentration from laser source to detector by our experimental result. It may also aid in designing the best model for oil chemistry, bio-pharmaceutical, laser medicine and application of medical engineering on LIF and coagulation in particle transport mode.

Rhodamine 6G Based New Fluorophore Chemosensor Toward Hg2+

  • Son, Young-A;Park, June-Min
    • Textile Coloration and Finishing
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    • v.24 no.3
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    • pp.158-164
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    • 2012
  • Rhodamine dyes belong to xanthene family has excellent photostability and photophysical properties. In rhodamine dyes, Rhodamine 6G and its precursors also have xanthene chromophore and it shows high fluorescent quantum yield. Rhodamine 6G derivates are simple to synthesis and its high sensitivity and water solubility are suitable as good chemosensor. In this regard, Rhodamine 6G derivates which have selectivity to specific metal cation can used to detect various heavy metal ions. In this study, rhodamine 6G derivatives were synthesized by reaction of rhodamine 6G hydrazide and glyoxal and 4-phenyl thiosemicarbazide and it showed colorimetric and fluorescence sensing toward $Hg^{2+}$ ion. This novel chemosensor was analyzed and measured on UV-Vis and fluorescence spectrophotometer. HOMO/LUMO values were also calculated by computational calculation.

QUANTITATION OF BARBITURATES IN URINE BY GC/MS AND ITS COMPARISON TO FLUORESCENCE POLARIZATION IMMUNOASSAY

  • Choo, Hea-Young;Park, Jeongeun;Park, Myung-Ja
    • Toxicological Research
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    • v.7 no.1
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    • pp.29-35
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    • 1991
  • Barbiturates are commonly abused tranquilizer and a rapid method to determine these drugs in biological samples is needed. In this study, was screened barbiturates in urine specimens by the fluorescence polarization immunoassay method(FPIA) and the positive samples were confirmed and identified by the more definitive GC/MS method. Fifteen positive smples which have barbiturate values higher than 0.5 ng/ml were analyzed by the GC/MS method. Eight samples were identified as phenobarbital and five samples were identified as crotilbarbitone.

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Effect of iron on the proteolytic activity of live Uronema marinum (Ciliata: Scoticociliatida) measured by fluorescence polarization

  • Lee, Eun-Hye;Kwon, Se-Ryeon;Choi, Seung-Hyuk;Kim, Ki-Hong
    • Journal of fish pathology
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    • v.19 no.1
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    • pp.83-86
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    • 2006
  • Effect of iron on the extracellular proteolytic activity of live Uronema marium was determined by fluorescence polarization (FP) method. Supplementation of 0.5 and 5.0 μM iron significantly increased caseinolytic activity of live U. marinum. In contrast, supplementation of 50 μM iron showed no significant differences in FP values compared to the control. The present result suggests that iron in cultured water or skin tissue of olive flounder may influence on the penetration and establishment of U. marinum, correlating with modulation of extracellular protease activity of the ciliates.

Formation of Reactive Oxygen Species and Cr(V) Entities in Chromium(VI) Exposed A549 Cells (크롬 6가 투여 후 A549 세포에서의 Reactive Oxygen Species와 크롬 5가의 발생)

  • 박형숙
    • Environmental Analysis Health and Toxicology
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    • v.11 no.1_2
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    • pp.49-57
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    • 1996
  • The production of reactive oxygen species on addition of hexavalent chromium (potassium dichromate, $K_2Cr_2O_7$ ) to lung cells in culture was studied using flow cytometer analysis. A Coulter Epics Profile flow cytometer was used to detect the formation of reactive oxygen species after $K_2Cr_2O_7$ was added to A549 cells grown to confluence. The cells were loaded with the dye, 2',7'-dichlorofluorescein diacetate, after which cellular esterases removed the acetate groups and the dye was trapped intracellularly. Reactive oxygen species oxidized the dye, with resultant fluorescence. Increased doses of Cr(VI) caused increasing fluorescence (10-fold higher than background at 200 gM). Addition of Cr(III) compounds, as the picolinate or chloride, caused no increased fluorescence. Electron paramagnetic resonance (EPR) spectroscopic studies indicated that three (as yet unidentified) spectral "signals" of the free radical type were formed on addition of 20, 50, 100 and 200 gM Cr(VI) to the A549 cells in suspension. Two other EPR 'signals" with the characteristics of Cr(V) entities were seen at field values lower than the standard free radical value. radical value.

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