• Title/Summary/Keyword: Intrinsic Fluorescence

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The Scattering and Absorption Effects of Fluorescence Spectroscopy in a Real Time Optical Biopsy (실시간 광학적 생검에서 형광분광법의 산란과 흡수에 대한 영향)

  • Han, Seunghee;Muller, Markus G.;Kang, Seunghee;Kang, Haejin
    • Progress in Medical Physics
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    • v.12 no.1
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    • pp.79-94
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    • 2001
  • The fluorescence emanating from a biological tissue contains information about scattering, absorption and the intrinsic fluorescence (fluorescence only due to fluorophores). Becaue fluorescence spectra of biological tissue are often significantly affected by the presence of tissue absorbers and scatterers, the measured tissue fluorescence cannot be interpreted as a linear combination of intrinsic fluorescence spectra of different tissue biochemical. We conducted experiments to examine the influence of scattering and absorption on the experimentally measured fluorescence of a turbid medium such as biological tissue. Therefore, we acquired fluorescence and reflectance spectra of tissue phantoms with a wide range of scatterer and absorber concentrations. By applying a photon migration model, which uses the scattering and absorption information contained in reflectance spectra to remove their distortion also present in fluorescence spectra, we extract the intrinsic fluorescence of these tissue models. We achieved excellent agreement between modeled and actual intrinsic fluorescence spectra. The motivation for this research is that intrinsic fluorescence spectra are expected to change with progression of disease in human tissue, due to changes in the tissue biochemical composition. It is not possible to distinguish the two tissue types by using only the measured fluorescence, however clear separation can be achieved with the intrinsic fluorescence in real time optical biopsy.

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Intrinsic UV Reflection and Fluorescence Studies for Water Sorption in Polycarbonate, Polyurethane and Poly(Ethylene Terephthalate) Films

  • Kim Min Sun;Sung Chong Sook Paik
    • Fibers and Polymers
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    • v.6 no.2
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    • pp.127-130
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    • 2005
  • Intrinsic UV reflection and fluorescence behaviors of polycarbonate, polyurethane and poly(ethylene terephthalate) films were investigated in order to characterize the interaction of water in these films. During water sorption process, UV reflection spectra of polycarbonate and polyurethane films showed little peak position changes. Fluorescence emission spectra of polycarbonate films showed red spectral shifts from 332 nm with water immersion time. This red-shifted peak could be due to phenyl-2-phenoxybenzoate, which is one of the major thermal degradation products in polycarbonate. Fluorescence peaks of polyurethane films appeared at two different positions and the ratio of these peak intensities increased with increasing immersion time. In the case of PET films, the UV reflection spectrum showed the peak intensity around 340 nm to change in response to water sorption. The fluorescence near 388 nm probably due to ground state dimer exhibited sensitivity with water sorption, when excited at 340 nm.

A Linear Beacon System Featuring an Internal Deoxyguanine Quencher Allows Highly Selective Detection of Single Base Mismatches

  • Lee, Young-Ae;Hwang, Gil-Tae
    • Bulletin of the Korean Chemical Society
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    • v.31 no.7
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    • pp.2011-2014
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    • 2010
  • The fluorescence intensity of a single-stranded oligonucleotide containing a fluorene-labeled deoxyuridine $(U^{Fl})$ unit increases by only 1.5-fold upon formation of its perfectly matched duplex. To increase the fluorescence signal during hybridization, we positioned a quencher strand containing a deoxyguanine (dG) nucleobase, functioning as an internal quencher, opposite to the $U^{Fl}$ unit to reduce the intrinsic fluorescence upon hybridization with a probe. From an investigation of the optimal length of the quencher strand and the effect of the neighboring base sequence, we found that a short strand (five-nucleotide) containing all natural nucleotides and dG as an internal quencher was effective at reducing the intrinsic fluorescence of a linear beacon; it also exhibited high total discrimination factors for the formation of perfectly matched and single base-mismatched duplexes. Such assays that function based on clear changes in fluorescence in response to single-base nucleotide mutations would be useful tools for accelerating diagnoses related to various diseases.

Effect of Ganglioside $G_{M3}$ on the Erythrocyte Glucose Transporter (GLUT1): Conformational Changes Measured by Steady-State and Time-Resolved Fluorescence Spectroscopy

  • Yoon, Hae-Jung;Lee, Min-Yung;Jhon, GiI-Ja
    • BMB Reports
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    • v.30 no.4
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    • pp.240-245
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    • 1997
  • Interactions between ganglioside $G_{M3}$ and glucose transporter, GLUT1 were studied by measuring the effect of $G_{M3}$ on steady-state and time-resolved fluorescence of purified GLUT1 in synthetic lipids and on the 3-O-methylglucose uptake by human erythrocytes. The intrinsic tryptophan fluorescence showed a GLUT 1 emission maximum of 335 nm, and increased in the presence of $G_{M3}$ by 12% without shifting the emission maximum, The fluorescence lifetimes of intrinsic tryptophan on GLUT1 consisted of a long component of 7.8 ns and a short component of 2,3 ns and $G_{M3}$ increased both lifetime components. Lifetime components were quenched by acrylamide and KI. Acrylarnide-mduced quenching of long-lifetime components was partly recovered by $G_{M3}$ However. KI-induccd quenching of short- and long-lifetime components was not rescued by $G_{M3}$. The anisotropy of 1.6-diphenyl-1.3.5-hexatriene (DPH)-probed dimyristoylphosphatidylcholine (DMPC) model membrane was also increased with $G_{M3}$ incorporation, The transport rate of 3-O-methylglucose increased by 20% with $G_{M3}$ incorporation on the erythrocytes, Therefore, $G_{M3}$ altered the environment of lipid membrane and induced the conformational change of GLUT1.

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Fluorescence of Tongue Coat - Mechanism of Tongue Coat Fluorescence and Suggestions on Development of Digital Tongue Diagnosis System - (설태의 형광특성 - 설태 형광현상의 발현기전 소개 및 제안 -)

  • Kim, Ji-Hye;Nam, Dong-Hyun
    • The Journal of the Society of Korean Medicine Diagnostics
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    • v.15 no.1
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    • pp.47-54
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    • 2011
  • In traditional Korean medicine, inspection of the tongue is an important method of making medical diagnoses and determining prognosis. We surveyed the fluorescence characteristics of the tongue coat in the ultraviolet light. The tongue coat comprises micro-organisms, blood metabolites, leukocytes from periodontal pockets, large amounts of desquamated epithelial cells released from the oral mucosa and different nutrients. In the ultraviolet light tissues of the oral cavity generally emit weak red or green fluorescence, which is not easily seen by the human eye, but is readily detected. This fluorescence has been proved to be due to the production of porphyrins by oral micro-organisms. While the composition of motile micro-organisms on the dorsum of the tongue is not constant, variations also occur persistingly in the fluorescence characteristics of the tongue coat. But because live bacteria contain a variety of intracellular biomolecules that have specific excitation and emission wavelength spectra characterizing their intrinsic fluorescence, the tongue coat emits fluorescence. the tongue itself, on the other hand, emits very weak or not fluorescence. In conclusion, we suggests that the uncoated tongue area be eliminated from the coated tongue area with the difference between the fluorescence characteristics of the tongue and that of the tongue coat.

Oligomerization State of the Plasma Membrane Proteolipid Apoprotein Purified from the Bovine Kidney, Probed by the Fluorescence Polarization

  • Chae, Quae;Nam, Sang-Rye
    • Bulletin of the Korean Chemical Society
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    • v.9 no.4
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    • pp.202-206
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    • 1988
  • In order to investigate the oligomerization state of the plasma membrane proteolipid apoprotein purified from the bovine kidney, fluorescence polarization experiment was carried out in the two different solvent systems, i.e., water and organic solvent(chloroform-methanol). The molecular volumes of the proteins estimated from the Perrin equation, were to be 45,258$A^3$ and 17,608$A^3$ in water and organic solvent, respectively. These values indicate that a trimerization is possibly occurring in the aqueous environment. As an auxiliary experiment for the calculation of the molecular volume using Perrin equation, fluorescence quenching constants ($K_q$) with the quencher acrylamide and fluorescence lifetimes (${\tau}_F$) of the intrinsic fluorophore tryptophan residue were estimated in the two different solvent systems. $K_q$ in water was 18.21$M^{-1}$ and it was 46.24$M^{-1}$ in organic solvent. Fluorescence lifetimes of tryptophan residue were calculated to be 2.80 nsec. in water and 3.81 nsec. in organic solvent, respectively.

Spectroscopic Characterization and Seasonal Distribution of Aquatic Humic Substances Isolated from Han River Water (한강원수로부터 분리된 수중휴믹물질의 계절적 분포와 분광학적 특성분석)

  • Kim, Hyun-Chul;Lee, Seock-Heon;Kim, Kyung-Ju;Yu, Myong-Jin
    • Journal of Korean Society of Environmental Engineers
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    • v.29 no.5
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    • pp.540-547
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    • 2007
  • Humic substances(HS) from Han River water was physic-chemically isolated by fractionational methods to investigate the seasonal distribution and to characterize the properties compared with intrinsic humic materials. Various HS samples were analyzed by element, Fourier transform infrared(FT-IR), proton nuclear magnetic resonance$(^1H-NMR)$ and fluorescence analyzers. The portion of HS from Han River water(HRHS) was 47.0% on the average, however it appeared that rainfall event brought about higher fraction of HS in Han River water by the periodic investigation. Aromaticity and humification degree of the HRHS were relatively lower than those of intrinsic humic materials originated from decomposing vegetation. FT-IR, $^1H-NMR$ and fluorescence spectroscopy showed the distinct differences between HRHS and intrinsic humic materials. Commercial humic materials could not represent structural and functional characteristics of local HS. The fluorescence spectroscopy, a relatively simple measurement, was found most useful tool to estimate humification degree for humic materials from particular sources.

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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Investigation of effects of newly synthesized Pt(II) complex against human serum albumin and leukemia cell line of K562

  • Divsalar, Adeleh;Saboury, Ali A.;Ahadi, Leila;Zemanatiyar, Elham;Mansouri-Torshizi, Hassan
    • BMB Reports
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    • v.43 no.11
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    • pp.766-771
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    • 2010
  • The biological evaluation of a new synthesized Pt(II)-complex, 2,2'-bipyridin Butylglycinato Pt(II) nitrate, an anti-tumor component, was studied at different temperatures by fluorescence and far UV circular dichroism (CD) spectroscopic methods. Human serum albumin (HSA) and human tumor cell line K562 were as targets. The Pt(II)-complex has a strong ability to quench the intrinsic fluorescence of HSA. Binding and thermodynamic parameters of the interaction were calculated by fluorescence quenching method. Far-UV-CD results showed that Pt(II)-complex induced increasing in content of $\alpha$ helical structure of the protein and stabilized it. The 50% cytotoxic concentration ($Cc_{50}$) of complex was determined using MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide) assay at different incubation times. Also, fluorescence staining with DAPI (4,6-diamidino-2-phenylindole) revealed some typical nuclear changes, which are characteristic of apoptosis. Above results suggest that Pt (II) complex is a promising anti-proliferative agent and should execute its biological effects by inducing apoptosis.

Fluorescence Characteristics of a Tryptophan Mutant of Leucine-responsive Regulatory Protein (Lrp) (트립토판 돌연변이 루신-반응 조절 단백질의 형광 특성)

  • Pokoo, Robert;Lee, Eui Ho;Lee, Chan Yong
    • Korean Journal of Microbiology
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    • v.50 no.4
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    • pp.275-280
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    • 2014
  • Leucine-responsive Regulatory Protein (Lrp) from Escherichia coli is an 18.8 kDa protein composed of 164 amino acids. Wild type Lrp (Lrp Wt) does not possess any tryptophan amino acid which has strong intrinsic fluorescence, whereas the mutant Lrp R145W contains a single tryptophan at the position 145 in the leucine-responsive domain. To investigate the fluorescence character, the Lrp R145W and Lrp Wt proteins were purified. The fluorescence intensity of Lrp R145W is much higher than that of wild type protein, and the intensity of Lrp R145W was decreased by binding to its specific DNA designed from ilvIH operon and to L-leucine. In addition, the tryptophan fluorescence intensity of Lrp R145W was strongly quenched by addition of acrylamide even in the least amount of concentration as well as by urea. The data obtained from this study may give valuable information on the three dimensional structure of Lrp R145W.