• Title/Summary/Keyword: Fluorescence method

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Caspase-11 Promoter-GFP Construct as a Dual Reporter of Cytotoxicity and Inflammation

  • Shin, Ki-Soon;Kang, Shin-Jung
    • Animal cells and systems
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    • v.10 no.2
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    • pp.73-77
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    • 2006
  • Caspase-11 has been known as a dual regulator of apoptosis and inflammatory response. An unusual feature of caspase-11 is that its expression is induced by apoptotic or proinflammatory stimuli. Utilizing these unusual features of caspase-11, we have developed a simple and sensitive assay method to screen pro- or anti-apoptotic/inflammatory molecules. To develop this assay method, we generated a reporter construct where GFP expression is regulated by caspase-11 promoter. When several types of cultured cells were transfected with this reporter construct and subsequently treated with various apoptotic or proinflammatory molecules, expression of GFP by the activation of caspase-11 promoter was easily detected by fluorescence microscopy or spectrofluorometry. In addition, a reduction of the GFP fluorescence was detected when an agent reported to suppress caspase-11 induction was applied. These results suggest that our reporter system can be used to screen pro- or anti-apoptotic/inflammatory molecules.

Spectrofluorimeteric Determination of Oxalate Based on Its Ternary Complex Between Eu3+ and Thenoyltrifluoroacetone

  • Cha, Ki-Won;Huang, Hua-Zi;Choi, Hyun Cheol
    • Bulletin of the Korean Chemical Society
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    • v.23 no.10
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    • pp.1456-1458
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    • 2002
  • A simple, sensitive and selective determination method of oxalate has been investigated based on the fluorescence enhancement of $Eu^{3+}$-TTA complex due to the formation of $Eu^{3+}$-TTA-oxalate ternary complex. An emission peak of $Eu^{3+}$-TTA, which is increased linearly with addition of oxalate, occurs at 610 nm in aqueous solution with excitation at 306 nm. The linear range of the calibration curve is 1 ${\times}$$10^{-6}$-8 ${\times}$$10^{-6}$ M and the detection limit is 1 ${\times}$$10^{-6}$ M. The effects of foreign ions were studied. The present method was applied to determine oxalate of two synthetic samples.

Fluorimetric Determination of Iron with N,N'-Oxalylbis(2-pyridyl-3'-sulphobenzoylhydrazone) (N,N'-Oxalylbis(2-pyridyl-3'-sulphobenzoylhydrazone)을 이용한 철의 형광광도법 정량)

  • Bark Ki-Min;Kim Yong-Nam
    • Journal of the Korean Chemical Society
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    • v.37 no.6
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    • pp.585-589
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    • 1993
  • A fluorimetric method for the determination of Fe(Ⅲ) using N,N'-oxalylbis(2-pyridyl-3'-sulphobenzoyl hydrazone)(OPSH) as a emission reagent has been developed. Determination has been performed by measuring the fluorescence intensity of Fe(Ⅲ) complex at 367 nm in aqueous solution (pH 3) with 290 nm excitation. There was a linear relationship between Fe(Ⅲ) concentration and fluorescence intensity over the range 2000∼10 ng/ml. The method has been applied to the determination of Fe(Ⅲ) in synthetic mixtures and tap water

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Application of Organized Media for Rapid Spectrofluorimetric Determination of Trace Amounts of Cr(VI) in the Presence of Cr(III)

  • Madrakian, Tayyebeh;Afkhami, Abbas;Mohammadnejad, Masoumeh
    • Bulletin of the Korean Chemical Society
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    • v.30 no.6
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    • pp.1252-1256
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    • 2009
  • A simple, selective and sensitive fluorescence quenching method was developed to the determination Cr(VI). The method is based on the oxidation of $I^-\;to\;{{I_3}^-}$ by Cr(VI) in sulfuric acid solution followed by immediate formation of ion association compound between I3 − and rhodamine 6G in Tween-80 micellar media at room temperature. The influence of several surfactants on rhodamine 6G fluorescence signal was studied; particular attention was paid in the aggregation behavior of rhodamine 6G–Tween-80 system. The experimental parameters (e.g., type of surfactant, reagent concentration) were studied and the optimal conditions were established. The linear calibration graph was obtained in the range 2.0 - 100.0 ng m$L^{-1}$ Cr(VI). The detection limit of the method was 0.37 ng m$L^{-1}$. The relative standard deviation (R.S.D.) is less than 5% (n = 5). The efficiency of the method for the determination of Cr(VI) in the presence of Cr(III) in the sample was investigated. The method was applied successfully to the determination of Cr(VI) and total Cr in water, and liver tissue samples.

Analysis of Chromosome aberrations by fluorescence in situ hybridization using triple chromosome-specific probes in human lymphocyte exposed to radiation (3중 DNA probe를 이용한 FISH(fluorescence in situ hybridization) 기법으로 방사선에 의한 염색체 이상 분석)

  • Chung, Hai-Won;Kim, Su-Young;Ha, Sung-Whan
    • Journal of Radiation Protection and Research
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    • v.24 no.1
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    • pp.45-53
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    • 1999
  • Fluorescence in situ hybridization with chromosome-specific probe has been shown to be a valid and rapid method for detection of chromosome rearrangements induced by radiation. This method is useful for quantifying structural aberrations, expecially for stable ones, such as translocation and insertion, which are difficult to detect with conventional method in human lymphocyte. In order to apply FISH method for high dose biological dosimetry, chromosomal abberations by radiation at doses of 1, 3, 5, and 7Gy were analysed with whole chromosome-specific probes by human chromosome 1, 2 and 4 according to PAINT system. The frequencies of stable translocation per cell equivalent were 0.04, 0.33, 1.22, 2.62, and 5.58 for the lymphocyte exposed to 0, 1, 3, 5, and 7Gy, respectively, and those of dicentric were 0.00, 0.06, 0.52, 1.19 and 2.44, respectively. Significantly more translocation of t(Ab), a translocated chromosome with a piece of painted acentric matrial 'b' attached to unpainted piece containing centromere 'A', than reciprocal chromosome t(Ba) was observed. The frequencies of all type of chromosome rearrangements increased with dose. From above result, FISH seemed to be useful for radiation biodosimetry by which the frequencies of various types of stable aberrations in human lymphocyte can be observed more easily than by conventional method and so will improve our ability to perform meaningful biodosimetry.

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COMPARATIVE STUDY ON THE EFFICACY OF DIGITAL IMAGING FIBER-OPTIC TRANS-ILLUMINATION AND LASER FLUORESCENCE IN MONITORING THE REMINERALIZATION PROCESS OF INCIPIENT SMOOTH SURFACE ENAMEL LESIONS (Digital Imaging Fiber-Optic Trans-Illumination과 Laser Fluorescence를 이용한 평활면 초기우식증의 재광화에 관한 비교 연구)

  • Hwang, Kyu-Seon;Yoo, Seung-Hoon;Kim, Jong-Soo
    • Journal of the korean academy of Pediatric Dentistry
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    • v.34 no.2
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    • pp.183-191
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    • 2007
  • Through out the world dental caries seems to be decreased as it is difficult to make an accurate diagnosis for dental caries. The traditional diagnostic method which is probing and x-ray taking has many limitations to diagnose the early caries, so there were recommendations for the needs of new equipments such as laser fluorescence(LF), digital imaging fiber-optic trans-illumination(DIFOTI), and quantitative light fluorescence (QLF) which were developed from various study results. Also confocal laser scanning microscopy(CLSM) and ultrasonics are used for research progression. This study is to evaluate whether it is possible to monitor accurately for remineralization amount of enamel surface early caries using DIFOTI or LF After inducing artificial caries to bovine teeth to 10 participants remineralization was enhanced by 0 ppm and 500 ppm fluoride mouth rinse solution for 3 weeks. Then they were cross sectioned and analyzed using gold standard of the lesion depth measured by CLSM. The following results were obtained: 1. The measured percentage of light intensity(luminosity ratio) by DIFOTI increased with remineralization period, and showed significant reverse correlation with lesion depth measured by CLSM (p<0.01). 2. The measurement of laser fluorescence increased with remineralization period, and showed significant correlation with lesion depth measured by CLSM (p<0.01). 3. To the result for CLSM, 500 ppm fluoride mouth rinse group showed rapid rate for decreased tendency of lesion depth than 0 ppm fluoride mouth rinse group. In conclusion DIFOTI system was used to measure accurately for the remineralization amount of early surface caries, it is a very useful equipment to detect precisely the changes for early enamel caries remineralization during treatments.

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DETECTION OF OCCLUSAL CARIES USING LASER FLUORESCENCE (레이저 형광법의 교합면 우식증 탐지 효과)

  • Kim, Chang-Gi;Lee, Chang-Seop;Lee, Sang-Ho
    • Journal of the korean academy of Pediatric Dentistry
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    • v.29 no.4
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    • pp.600-606
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    • 2002
  • The purpose of this study was to evaluate the diagnostic validity of an incipient occlusal caries using argon laser fluorescence. Extracted human premolars and molars with enamel carious lesion of occlusal surface were assessed using visual examination, visual examination with probing, argon laser fluorescence and histologic depth of carious lesion. The results in each of all the three detection methods were compared to the assessment of histologic depth of carious lesion using polarized microscope. The results from the present study can be summarized as follows; 1. There was highly correlation between the histologic depth of occlusal caries and all three detection methods(P<0.01). 2. The reproducibility(kappa value) of the visual examination, visual examination with probing and argon laser fluorescence between the histologic depth of occlusal caries was 0.189, 0.128, 0.472. The highest correlation was seen between detection of occlusal caries by argon laser fluorescence and histologic scores by polarized microscope. The results from this study indicated that argon laser fluorescence considered to be accurate and reliable method in detecting occlusal caries.

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Fluorescence Anisotropy Study on the Effect of Phellodendri Cortex's Berberine on Regulation of the Function of DNA (황백(黃柏)의 berberine이 DNA의 기능조절에 미치는 영향에 관한 형광이방성 연구)

  • Lee, Seong Kyung;Han, Hyo Sang;Huh, Sung Ho
    • The Korea Journal of Herbology
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    • v.33 no.5
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    • pp.105-110
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    • 2018
  • Objectives : We tried to observe the fluorescence anisotropy and intensity of ethidium ion in the intercalating binding interaction between DNA and ethidium ions in the presence of berberine, and then tried to explain the effect of berberine on the intercalating interaction of ethidium ion with DNA. Methods : DNA(calf thymus DNA), berberine and ethidium bromide(EtBr) were purchased from Sigma-Aldrich Co. Proper amount of each compound was dissolved in 20 mM sodium phosphate buffer(pH 7.0) containing 100 mM of NaCl to prepare stock solutions. Collections of the fluorescence anisotropy and intensity data were performed on JASCO FP-8300 spectrofluorometer equipped with a polarizer and a Peltier temperature controller. The excitation of ethidium ion was done at 550 nm and the emission data were collected at 600 nm. For Stern-Volmer plot, the fluorescence data were collected at $18^{\circ}C$ and $30^{\circ}C$. Results : According to the results of this research, the weak competitive binding pattern between ethidium ion and berberine appeared in binding with DNA at low ratio of DNA to ethidium ion. But at high ratio of DNA to ethidium ion, this weak competition disappeared. Instead, berberine might bind to DNA by intercalating way. In other words, berberine could de-intercalate ethidium ion from DNA at low concentration of DNA relative to ethidium ion, but could not at high concentration of DNA relative to ethidium ion. In addition, the mechanism of fluorescence quenching of ethidium ion could also proceed differently, depending on the ratio of the amount of DNA to that of ethidium ion. Conclusions : The effect of berberine on the DNA-ethidium ion intercalating interaction could work differently, depending on the relative ratio of the amount of DNA to that of ethidium ion. This study also showed that fluorescence anisotropy analysis is very useful method to obtain detailed information for investigation of the complex binding interactions. In order to fully understand the mechanism of action of the pharmacological effect by berberine, studies on the effect of berberine on the action of proteins such as various enzymes closely related to berberine-induced medicinal effects should be continued.