• Title/Summary/Keyword: fluorescent probe technique

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Ibuprofenlysine binding to human and bovine serum albumin using a fluorescence probe technique

  • Kim, Chong-Kook;Cha, Hyun-Sook;Kim, Yang-Bae;Yu, Byung-Sul
    • Archives of Pharmacal Research
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    • v.4 no.1
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    • pp.19-24
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    • 1981
  • The possibility of using a fluorescence probe technique for the study of ibuprofenlysine binding to human and bovine serum albumin was investigated. 1-anilino-8-naphalenesulfonate was used as the probe. The number of binding sites of human and bovine serum albumins for ibuprofenlysine appears to be 4 and 2, respectively. By using this technique, the association constants were found to be $1.533{\times}10^{4}M^{-1}$ and $2.238{\times}10^{4}M^{-1}$, respectively.

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The Effect of Cure History on the Fluorescence Behavior of an Unsaturated Polyester Resin with A Fluorescence Probe

  • Donghwan Cho;Yun, Suk-Hyang;Bang, Dae-Suk;Park, Il-Hyun
    • Macromolecular Research
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    • v.12 no.3
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    • pp.282-289
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    • 2004
  • We have extensively characterized the fluorescence behavior of unsaturated polyester (UP) resin in the absence and presence of a 1,3-bis-(l-pyrenyl)propane (BPP) fluorescent probe at various dynamic and isothermal cure histories by means of a steady-state fluorescence technique using a front-face illumination equipment. In addition, we explored the effect of the fluorescence intensity on the relaxation of the fluorescent probe in the UP resin by resting the dynamically and isothermally cured resin at ambient temperature and pressure for 24 h. The monomer fluorescence intensity, which has two characteristic peaks at 376 and 396nm, changed noticeably depending on the cure temperature and time and provided important information with respect to the molecular and photophysical responses upon curing. The result of the fluorescence study indicates that the increased local viscosity and restricted molecular mobility of the UP resin surrounding the BPP probe after curing are both responsible for the enhancement of the monomer fluorescence intensity. Our results also demonstrate that once the BPP probe has enough time to rearrange and become isolated prior to fluorescence, a sufficient amount of fluorescence is emitted. Therefore, we note that the fluorescence behavior of this UP resin system is influenced strongly by the relaxation process of the fluorescent probe in the resin as well as process used to cure the resin.

The Region of Distribution of Barbiturates in Synaptosomal Plasma Membrane Vesicles Isolated from Rat Brain as Studied by Fluorescence Quenching (Barbiturates가 생체세포막 외측 단층의 소수성 부위와 친수성 부위에 분포되는 상대적 비율)

  • Yun, Il;Lee, Byung-Woo
    • 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.

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Development of FISH Methods for Detection of Genus Bacillus (Bacillus 속 세균을 검출하기 위한 Fluorescent In Situ Hybridization 방법의 개발)

  • 홍선희;김옥선;송홍규;이동훈;안태석
    • Korean Journal of Microbiology
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    • v.37 no.3
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    • pp.204-208
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    • 2001
  • A technique for detection of Bacillus in soil and waste water treatment system was developed. Mixed col- tured solutions of 22 Bacillus strains were applied for selection of probe and pretreatment method for FISHAmong the three probes known as useful tool for FISH method,S-G-Bacill-0597-a-A-22 was best for detec-tion of Bacillus. Ninety five percent of DAPl count was observed with FISH method with S-G-Bacill-0597-a-A-22 probe. For increasing the permeability to Bacillus cell walls, pretreatment with lysozyme was betterthan that with lysozyme and SDS/DTT (dithiothreitol). Bacillus spore was not detected with FISH. So, Bacil-lus detection in ecosystem requires FISH with pretreatment of lysozyme and spore staining.

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The Effect of n-Alkanols on the Lateral Diffusion of Synaptosomal Plasma Membrane Vesicles Isolated from Bovine Cerebral Cortex (n-Alkanols가 소의 대뇌피질로부터 분리한 Synaptosomal Plasma Membrane Vesicles의 측방확산운동 범위와 속도에 미치는 영향)

  • Chung, In-Kyo;Kang, Jung-Sook;Yun, Il
    • The Korean Journal of Pharmacology
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    • v.29 no.1
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    • pp.157-163
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    • 1993
  • Intramolecular excimer formation with the fluorescent probe 1,3-di(1-pyrenyl)propane (Py-3-Py) was used to investigate the effects of methanol, ethanol, 1-propanol, 1-butanol, 1-pentanol, 1-hexanol, 1-heptanol, 1-octanol, 1-nonanol and 1-decanol on the lateral diffusion of synaptosomal plasma membrane vesicles isolated from bovine cerebral cortex (SPMV). The n-alkanols increased the excimer to monomer fluorescence intensity ratio (I'/I) of Py-3-Py in the SPMV. In a dose-dependent manner, n-alkanols increased lateral diffusion of hydrocarbon region of bulk (inner+outer monolayers) SPMV lipid bilayers, and the potencies of n-alkanols up to l-nonanol increased with carbon chain length. It appears that the potencies in bilayer fluidization due to the lateral diffusion increase by 1 order of magnitude as the carbon chain length increases by two carbon atoms. The cut-off phenomenon was reached at 1-decanol, where further increase in hydrocarbon length resulted in a decrease in pharmacological activity.

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Seasonal Changes of bacterial community analysed by fluorescent in situ hybridization method in Lake Soyang (Fluorescent In Situ Hybridization방법으로 분석한 소양호 세균 군집 구조의 계절적 변화)

  • Hong, Sun-Hee;Ahn, Tae-Seok
    • Korean Journal of Microbiology
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    • v.34 no.3
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    • pp.169-174
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    • 1998
  • To define the structure and diversity of bacterial communities in the aqutic ecosystem, Lake Soyang, the largest artificial reservoir in Korea, a new method, fluorescent in situ hybridization was applied. This technique relies on the specific hybridization of the nucleic acid probes to the naturally amplified intracellular rRNA. By this method, the bacterial community composition of Lake Soyang and bacterial numbers belong to eubacteria, proteobacteria and Cytophaga-Flavobacterium group were estimated. Total bacterial numbers ranged from $0.3{\times}10^6{\sim}2.0{\times}10^6cells{\cdot}ml^{-1}$, and vertical profile of total bacteria showed the peak at 2 and 5 m depths. The ratio of eubacteria to total bacteria were 22~100% and varied with depth and season. The percentage of Proteobacteria ${\alpha}$-group ranged 2.6~66.7%, ${\beta}$-group 4.5~53.5%, ${\gamma}$-group 4.6~76.7% and Cytophaga-Flavobacterium group 2.1~35.9%. Also, bacteria] community had spatial and temporal characteristics. The dominant groups were ${\beta}$-group in winter, ${\gamma}$-group in spring and early summer and ${\alpha}$-group in summer.

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Characterization of the Surface Contribution to Fluorescence Correlation Spectroscopy Measurements

  • Chowdhury, Salina A.;Lim, Man-Ho
    • Bulletin of the Korean Chemical Society
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    • v.32 no.2
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    • pp.583-589
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    • 2011
  • Fluorescence correlation spectroscopy (FCS) is a sophisticated and an accurate analytical technique used to study the diffusion of molecules in a solution at the single-molecule level. FCS is strongly affected by many factors such as the stability of the excitation power, photochemical processes, mismatch between the refractive indices, and variations in the cover glass thickness. We have studied FCS near the surface of a cover glass by using rhodamine 123 as a fluorescent probe and have observed that the surface has a strong influence on the measurements. The temporal autocorrelation of FCS decays with two characteristic times when the confocal detection volume is positioned near the surface of the cover glass. As the position of the detection volume is moved away from the surface, the FCS autocorrelation becomes one-component decaying; the characteristic time of the decay is the same as the faster-decaying component in the FCS autocorrelation near the surface. This observation suggests that the faster component can be attributed to the free diffusion of the probe molecules in the solution, while the slow component has its origin from the interaction between the probe molecules and the surface. We have characterized the surface contribution to the FCS measurements near the surface by changing the position of the detection volume relative to the surface. The influence of the surface on the diffusion of the probe molecules was monitored by changing the chemical properties of the surface. The surface contribution to the temporal autocorrelation of the FCS strongly depends on the chemical nature of the surface. The hydrophobicity of the surface is a major factor determining the surface influence on the free diffusion of the probe molecules near the surface.

Gene Expression in Zn-deficient U937 Cell Line : Using cDNA Microarray (아연결핍된 단핵구 U937 Cell Line에 있어서의 유전자 발현 탐색 : cDNA Microarray 기법 이용)

  • Beattie, John H.;Trayhurn, Paul
    • Journal of Nutrition and Health
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    • v.35 no.10
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    • pp.1053-1059
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    • 2002
  • In post-genome period, the technique for identifying gene expression has been changed to high throughput screening. In the field of molecular nutrition, the need for this technique to clarify molecular function of the specific nutrient is essential. In this study, we have tested the zinc-regulated gene expression in zinc-deficient U937 cells, using cDNA microarray which is the cutting-edge technique to screen large numbers of gene expression simultaneously. The study result can be used for the preliminary gene screening data for clarifying, using monocyte U937 cell line, molecular Zn aspect in atherosclerosis. U937 cells were cultured in Zn-adequate (control, 12 $\mu$M Zn) or Zn-deficient (experimental, 0 $\mu$M Zn) ESMI media during 2 days, respectively. Cells were harvested and RNA was extracted. Total RNA was reverse-transcriptinized and synthesized cDNA probe labeled with Cy-3. fluorescent labeled cDNA probe was applied to microarray slide for hybridization slide, and after then, the slide was scanned using fluorescence scanner. ‘Highly expressed genes’ in Zn-deficient U937 cells, comparing to Zn-adequate group, are mainly about the genes for motility protein, immune system protein, oncogene and tumor suppressor and ‘Less highly expressed genes’ are about the genes for transcription, apoptosis associated protein, cell cycle, and several basic transcription factors. The results of this preliminary study imply the effectiveness of cDNA microarray for expression profiling of a singly nutrient deficiency, specially Zn. Furthur study, using tailored-cDNA array and capillary endothelial cell lines, would be beneficial to clarify molecular Zn function, more in detail.

The Distribution of Barbiturates in Model Membranes of Total Lipids and Total Phospholipids Extracted from Brain Membranes

  • Park, Chang-Sik;Lee, Seong-Moon;Chung, In-Kyo;Kim, Jin-Bom;Son, Woo-Sung;Jang, Hye-Ock;Yun, Il
    • BMB Reports
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    • v.33 no.3
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    • pp.221-227
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    • 2000
  • The distribution of barbiturates in the model membranes of total lipids (SPMVTL) and total phospholipids (SPMVPL) extracted from synaptosomal plasma membrane vesicles was determined by employing a fluorescent probe technique. The two fluorescent probes 2-(9-anthroyl)stearic acid and 12-(9-anthroyl)stearic acid were utilized as probes for the surface and the hydrocarbon interior of the outer monolayer of the SPMVTL and SPMVPL, respectively. The Stern-Volmer equation of fluorescent quenching was modified to calculate the relative distribution. The analysis of preferential quenching of these probes by barbiturates indicates that pentobarbital, hexobarbital, amobarbital and phenobarbital are predominantly distributed on the surface area, while thiopental sodium has an accessibility to the hydrocarbon interior of the outer monolayer of the SPMVTL and SPMVPL. 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 a higher general anesthetic activity.

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Assembly and electrical property of GFP/Cytochrome b562 Fusion Protein ontothe Au Substrate

  • Jeong, Seong-Cheol;Choe, Jeong-U;Lee, Won-Hong;Nagamune, T.
    • 한국생물공학회:학술대회논문집
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    • 2003.04a
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    • pp.630-633
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    • 2003
  • Transfer of an electron from one site to another in a molecular or between molecules and/or electrodes is one of the most fundamental and ubiquitous processes in chemistry, biology and physics. In this study fusion proteins composed by green fluorescent protein(GFP) and cytochrome b562 were used in fabricating molecular array as an electron sensitizer and electron acceptor, Protein formation onto the substrate was performed by the self-assembly technique. The fusion protein film were analyzed using scanning probe microscope(SPM), Surface Plasmon Resornance(SPR) and hybrid STM/I-V. The results suggest that the proposed molecular photodiode can be used as a basic unit of the memory device.

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