• Title/Summary/Keyword: Fluorescence probe

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Synthesis and characterization of ABA types tri-block copolymers derived from p-dioxanone, ${\varepsilon}-caprolactone$ and poly(ethylene glycol)

  • Remant Bahadur K.C.;Bhattarai Shanta Raj;Aryal Santosh;Khil, Myung-Seob;Kim, Hak-Yong;Lee, Douk-Rae
    • Proceedings of the Polymer Society of Korea Conference
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    • 2006.10a
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    • pp.255-255
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    • 2006
  • Polymeric nanoparticles are recognized as promising drug carriers [1]. Here, novel tri-block copolymers based on poly PPDO, PCL and PEG were synthesized and employed for the formulation of reproducible polymeric nanoparticles [2]. To estimate the feasibility of the polymer to form polymeric nanoparticles, nanoparticles were prepared by co-solvent evaporation technique. Polymerization and structural features of the polymer were analyzed by different physico-chemical techniques. Existence of hydrophobic domains as a core of nanoparticles was characterized by $^{1}H-NMR$ spectroscopy, and further confirmed by fluorescence technique using pyrene as probe.

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Immobilization of Proteins on Magnetic Nanoparticles

  • Wang, Tzu-Hsien;Lee, Wen-Chien
    • Biotechnology and Bioprocess Engineering:BBE
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    • v.8 no.4
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    • pp.263-267
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    • 2003
  • Magnetic nanoparticles prepared from an alkaline solution of divalent and trivalent iron ions could covalently bind protein via the activation of Nethyl-N-(3-dimethylaminopropyl) carbodiimide (EDC). Trypsin and avidin were taken as the model proteins for the formation of protein-nanoparticle conjugates. The immobilized yield of protein increased with molar ratio of EDC/nanoparticie. Higher concentrations of added protein could yield higher immobilized protein densities on the particles. In contrast to EDC, the yields of protein immobilization via the a ctivation of cyanamide were relatively lower. Nanoparticles bound with avidin could attach a single-stranded DNA through the avidin-biotin interaction and hybridize with a DNA probe. The DNA hybridization was confirmed by fluorescence microscopy observations. Immobilized DNA on nanoparticles by this technique may have widespread applicability to the detection of specific nucleic acid sequence and targeting of DNA to particular cells.

마이크로 어레이를 이용한 돼지 장염 바이러스의 신속한 감별 진단

  • 조호성;김현진;김용환;조경오;박남용
    • Proceedings of the Korean Society of Veterinary Pathology Conference
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    • 2002.11a
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    • pp.142-142
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    • 2002
  • 돼지의 주요 장염 유발 바이러스인 돼지 전염성 위장염 바이러스 (transmissible gastroenteritis virus; TGEV), 돼지 유행성 설사증 바이러스 (porcine epidemic diarrhea virus; PEDV), 돼지 칼리시 바이러스 (porcine enteric calicivirus; PECV), 돼지 로타바이러스 A 형과 C 형 (porcine rotavirus; PRY, group A and C)을 동시에 감별 진단 할 수 있는 신속하고 정확한 oligonucleotide microarray 진단법을 개발하였다. 이 진단법은 유리슬라이드에 각각의 바이러스에 특이적인 부위에서 제작된 oligonucleotide probe를 찍은 DNA chip을 제작하여 여기에 각각의 바이러스를 역전사하고 cy5-dCTP를 포함한 multiplex PCR을 수행한 다음 hybridization 하였다. 이후 hybridization 결과는 fluorescence scanner를 이용하여 확인하였다. 이 새로운 microarray system은 RT-PCR과 같은 기존의 진단방법보다 소량의 바이러스를 민감하게 검사할 수 있을 뿐 아니라 hybridization을 통해 검사결과의 정확성을 확인할 수 있었다. 따라서 본 연구에서 개발한 microarray system은 돼지의 설사 유발 바이러스를 진단하는데 매우 유용한 진단 방법임을 확인하였다.

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Modeling and Experimental Study of Radio-frequency Glow Discharges and Applications for Plasma Processing

  • Kang, Nam-Jun
    • Proceedings of the Korean Vacuum Society Conference
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    • 2012.02a
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    • pp.179-179
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    • 2012
  • Low pressure radio-frequency glow discharges are investigated using theoretical modeling and various experimental diagnostic methods. In the calculations, global models and transformer models are developed to understand the chemical kinetics as well as the electrical properties such as the effective collision frequency, the heating mechanism and the power transferred to the plasma electrons. In addition, Boltzmann equation solver is used to compensate the effect of the electron energy distribution function (EEDF) shape in the global model, and the general expression of energy balance for non-Maxwellian electrons is developed. In the experiments, a number of traditional plasma diagnostic methods are used to compare with calculated results such as Langmuir probe, optical emission spectroscopy (OES), optical absorption spectroscopy (OAS) and two-photon absorption laser-induced fluorescence (TALIF). These theoretical and experimental methods are applied to understand several interesting phenomena in low pressure ICP discharges. The chemical and physical properties of low pressure ICP discharges are described and the applications of these methods are discussed.

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Effect of Drug Substances on the Microviscosity of Lipid Bilayer of Liposomal Membrane

  • Han, Suk-Kyu;Kim, Jin-Suk;Lee, Yong-Soo;Kim, Min
    • Archives of Pharmacal Research
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    • v.13 no.2
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    • pp.192-197
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    • 1990
  • The microviscosites of the lipid bilayers of liposomal membranes of phospholipids were measured by the intermolecular excimer, formation method employing pyrene as a fluorescence probe, and the effects of n-alkanols and other local anesthetics on the microviscosity were investigated. The results showed that the n-alkanols and the ohter local anesthetics effectively lowered the microviscosity of the lipid bilayer of the dipalmitoyl phosphatidycholine liposomal membrane in proportion to the concentration of the additives. Moreover, there was a fairly good correlation between the ocal anesthetic activities and the microviscosity-lowering activities of these drugs. This results suggests that the nerve blocking activity of local anesthetics might have some relation with their activity fluidizing the lipid bilayer of biomembrane.

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Effects of n-Alkanols on the Lateral Diffusion of Total Phospholipid Fraction Extracted from Brain Membranes

  • Chung, In-Kyo;Kang, Jung-Sook;Yun, Il
    • Archives of Pharmacal Research
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    • v.16 no.3
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    • pp.191-195
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    • 1993
  • We investigated the effects of n-alkanols on the range and rate of the lateral diffusion of 1, 3-di(1-pyrenyl)propane in the model membranes of total phospholipid fraction extracted from synaptosomal plasma membrane vesicles. n-Akanols increased the range and rate of the lateral diffusion of 1, 3-di(1-pyrenyl)propane in the bulk model membrane structures (inner + outer monolayers) and the potencies of n-alknols up to 1-nonanol increased 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 icnrease in hydrocarbon length resulted in a decrease in the lateral diffusion. However, significant changes in the 1'/1 value were not observed by methanol (from 100 to 2500 mM), ethanol (from 25 to 800 mM), and 1-propanol (from 10 to 250 mM) over entire concentration.

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Identification of a Tandemly Repented DNA Sequence Using Combined RAPD and FISH in Welsh Onion (Allium fistulosum)

  • Bong Bo Seo;Geum Sook Do;Seon Hee Lee
    • Animal cells and systems
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    • v.3 no.1
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    • pp.69-72
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    • 1999
  • A tandemly repeated DNA sequence was identified and characterized y the combined RAPD and FISH data from a total genomic DNA of Welsh onion (Allium fistulosum). A clone containing this repeating sequence was selected and sequenced. This repeating unit of 314 bp inserted into pAf 072 contained 54.1% adenine and thymine residues, and showed the primer sequence used, 5'-GAAACGGGTG-3', in both terminals of the sequence. Fluorescence in situ hybridization using this tandemly repeated sequence as a probe indicated that the detected sites were coincident with the major C-banded constitutive heterochromatin in the terminal regions of both arms of all 6 chromosomes.

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Self-Assembled Peptide Structures for Efficient Water Oxidation

  • Lee, Jae Hun;Lee, Jung Ho;Park, Yong Sun;Nam, Ki Tae
    • Proceedings of the Korean Vacuum Society Conference
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    • 2013.08a
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    • pp.280-280
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    • 2013
  • In green plants, energy generation is accomplished through light-harvesting photosystem, which utilize abundant visible light and multi-stepwise redox reaction to oxidize water and reduce NADP+, transferring electrons efficiently with active cofactors1. Inspired by natural photosynthesis, artificial solar water-splitting devices are being designed variously. However, the several approaches involving immobilization2, conjugation3, and surface modification4 still have limitations. We have made artificial photosynthesis templates by self-assembling tyrosine-based peptide to mimick photosystem II. Porphyrin sensitizer absorbing blue light strongly was conjugated with the templates and they were hybridized with cobalt oxide through the reduction of cobalt ions in an aqueous solution. The formation of hybrid templates was characterized using TEM, and their water oxidation performance was measured by fluorescence oxygen probe. Our results suggest that the bio-templated assembly of functional compounds has a great potential for artificial photosynthesis.

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Effects of Barbiturates on Transbilayer Fluidity Domains of Phospholipid Model Membrane Monolayers (Barbiturates가 소의 대뇌피질 Synaptosomal Plasma Membrane Vesicles로 부터 추출 제제한 총지질 및 총인지질 인공세포막에 형성된 비대칭적 유동성에 미치는 비대칭적 영향)

  • Yun, Il;Kang, Jung-Sook
    • The Korean Journal of Pharmacology
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    • v.28 no.1
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    • pp.103-114
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    • 1992
  • Selective quenching of 1,6-diphenyl-1,3,5-hexatriene (DPH) by trinitrophenyl groups was utilized to examine the transbilayer fluidity domains of the model membranes of total lipids (SPMVTL) and phospholipids (SPMVPL) extracted from synaptosomal plasma membrane vesicles. At $37^{\circ}C$, all anisotropy (r), limiting anisotropy $(r_{\infty})$, and order parameter (S) values of DPH in the SPMVTL were larger than those in SPMVPL. The anisotropy, limiting anisotropy, and order parameter of DPH in the inner monolayer were 0.025, 0.033, and 0.070, respectively, greater than calculated for the outer monolayer of SPMVTL. In SPMVPL, the anisotropy, limiting anisotropy, and order parameter of DPH in the inner monolayer were 0.014, 0.018, and 0.047, respectively, greater than calculated for the outer monolayer. Selective quenching of DPH by trinitrophenyl groups was also utilized to examine the effects of barbiturates on the transbilayer fluidity domains of SPMVTL and SPMVPL. Barbiturates did not affect the anisotropy of DPH in the transbilayer domains of SPMVTL. In contrast, barbiturates increased the fluorescence anisotropy, limiting anisotropy, and order parameter of DPH in the SPMVPL in a dose-dependent manner. Barbiturates showed a greater ordering effect on the outer monolayer as compared to the inner monolayer of SPMVPL. Hence, it has been demonstrated for the first time that the Sheetz-Singer hypothesis (1974) may be valid for phospholipid model membranes.

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Detection of Mitochondrial ATP-Sensitive Potassium Channels in Rat Cardiomyocytes

  • Cuong, Dang Van;Kim, Na-Ri;Kim, Eui-Yong;Lee, Young-Suk;Kim, Hyun-Ju;Kang, Sung-Hyun;Hur, Dae-Young;Joo, Hyun;Park, Young-Shik;Hong, Yong-Geun;Lee, Sang-Kyung;Chung, Joon-Yong;Seog, Dae-Hyun;Han, Jin
    • The Korean Journal of Physiology and Pharmacology
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    • v.8 no.4
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    • pp.201-206
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    • 2004
  • Mitochondrial ATP-sensitive potassium $(mitoK_{ATP})$ channels play a role in early and late ischemic preconditioning. Nevertheless, the subunit composition of $mitoK_{ATP}$ channels remains unclear. In this study, we investigated the subunit composition of $mitoK_{ATP}$ channels in mitochondria isolated from rat cardiac myocytes. Mitochondria were visualized using the red fluorescence probe, Mitrotracker Red, while $mitoK_{ATP}$ channels were visualized using the green fluorescence probe, glibenclamide-BODIPY. The immunofluorescence confocal microscopy revealed the presence of Kir6.1, Kir6.2 and SUR2 present in the cardiac mitochondria. Western blot analysis was carried to further investigate the nature of $mitoK_{ATP}$ channels. For SUR proteins, a 140-kDa immunoreactive band that corresponded to SUR2, but no SUR1 was detected. For Kir6.2, three bands $({\sim}44,\;{\sim}46,\;and\;{\sim}30\;kDa)$ were detected, and a specific ${\sim}46-kDa$ immunoreactive band corresponding to Kir6.1 was also observed. These observations suggest that the subunits of $mitoK_{ATP}$ channels in rat myocytes include Kir6.1, Kir6.2, and a SUR2-related sulfonylurea-binding protein.