• 제목/요약/키워드: fluorescence life time

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Generation and Characterization of Cell-Permeable Greem Fluorescent Protein Mediated by the Basic Domain of Human Immunodeficiency Virus Type 1 Tat

  • Park, Jin-Seu;Kim, Kyeong-Ae;Ryu, Ji-Yoon;Choi, Eui-Yul;Lee, Kil-Soo;Choi, Soo-Young
    • Journal of Microbiology and Biotechnology
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    • 제10권6호
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    • pp.797-804
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    • 2000
  • The human immunodeficiency virus type 1 (HIV-1) Tat is one of the viral gene products essential for HIV replication. The exogenous Tat protein is transduced through the plasma membrane and then accumulated in a cell. The basic domain of the Tat protein, which is rich in arginine and lysine residues and called the protein transduction domain (PTD), has been identified to be responsible for this transduction activity. To better understand the nature of the transduction mediated by this highly basic domain of HIV-1 Tat, the Green Fluorescent Protein (GFP) was expressed and purified as a fusion protein with a peptide derived from the HIV-1 Tat basic domain in Escherichia coli. The transduction of Tat-GFP into mammalian cells was then determined by a Western blot analysis and fluorescence microscopy. The cells treated with Tat-GFP exhibited dose- and time-dependent increases in their intracellular level of the protein. the effective transduction of denatured Tat-GFP into both the nucleus and the cytoplasm of mammalian cells was also demonstrated, thereby indicating that the unfolding of the transduced protein is required for efficient transduction. Accordingly, the availability of recombinant Tat-GFP can facilitate the simple and specific identification of the protein transduction mediated by the HIV-1 Tat basic domain in living cells either by fluorescence microscopy or by a fluorescence-activated cell sorter analysis.

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COMPARISON OF PLASMA-INDUCED SURFACE DAMAGES IN VARIOUS PLASMA SOURCES

  • Yi, Dong-Hyen;Lee, Jun-Sik;Kim, Sang-Kyun;Kim, Jae-Jeong
    • 한국표면공학회지
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    • 제29권5호
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    • pp.338-344
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    • 1996
  • This study was an investigation of plasma-induced damages on silicon substrate in the semiconductor manufacturing technology. The plasma-induced damage level on silicon substrate was analyzed and compared in various plasma etching systems. The analysis methods were therma wave, life-time recovery, SCA (Surface Charge Analyzer) and TRXF (Total Reflection X-ray Fluorescence) measurements, and the measured values were compared for each systems. In the comparison of the values which were obtained by a system that had low life-time recovery, there was not any differences in DC parameters. However, the reflesh time distribution of device of that system had decreased about 10 to 20m sec compared to a system which had high life-time recovery.

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In Vivo Measurement of Plant Vitality by the Fluorescence Transient

  • Soonja Oh;Koh, Seok-Chan
    • 한국자원식물학회:학술대회논문집
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    • 한국자원식물학회 2002년도 제9차 국제심포지움 및 추계정기학술발표회
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    • pp.12-12
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    • 2002
  • The chlorophyll fluorescence combined with the O-J-I-P transients were examined in the leaves of the crinum plants (Crinum asiaticum var. japonicum BAK.), in order to satisfy the demand for rapid in vivo measurement of vitality, and to apply easily to approach questions of economical interest concerning the plant vitality. The photosynthetic efficiency, Fv/Fm, of crinum plants dramatically decreased depending on temperature drop in winter. In summer, the Fv/Fm values was lower in day time than at dawn and night, suggesting that photosynthetic efficiency is chronically photoinhibited in day time. In winter, there was no prominent diurnal fluctuations of Fv/Fm values. However, based on the O-J-I-P transient, PI$\_$NO/ and SFI$\_$NO/ dramatically increased at noon in summer, and $\psi$o/(1-$\psi$o) diurnally fluctuated in winter. These results indicated that vitality indexes such as PI$\_$NO/, SFI$\_$NO/ and $\psi$o/(1-$\psi$o) can be used as the indicators for in vivo measurement of environmental stresses.

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In Vivo Measurement of Plant Vitality by the Fluorescence Transient

  • Soonja Oh;Koh, Seok-Chan
    • 한국자원식물학회:학술대회논문집
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    • 한국자원식물학회 2002년도 심포지엄
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    • pp.88-95
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    • 2002
  • The chlorophyll fluorescence combined with the O-J-I-P transients were examined in the leaves of the crinum plants (Crinum asiaticum var.japonicum BAK.), in order to satisfy the demand for rapid in vivo measurement of vitality, and to apply easily to approach questions of economical interest concerning the plant vitality. The photosynthetic efficiency, Fv/Fm, of crinum plants dramatically decreased depending on temperature drop in winter. In summer, the Fv/Fm values was lower in day time than at dawn and night, suggesting that photosynthetic efficiency is chronically photoinhibited in day time. In winter, there was no prominent diurnal fluctuations of Fv/Fm values. However, based on the O-J-I-P transient, PI$\_$NO/ and SFI$\_$NO/ dramatically increased at noon in summer, and $\psi$ο/(1-$\psi$ο) diurnally fluctuated in winter. These results indicated that vitality indexes such as PI$\_$NO/, SFI$\_$NO/ and $\psi$ο/(1-$\psi$ο) can be used as the indicators for in vivo measurement of environmental stresses.

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플루오레신나트륨의 농도 범위 분석 (Analysis for Concentration Range of Fluorescein Sodium)

  • 이다애;김용재;윤기철;김광기
    • 대한의용생체공학회:의공학회지
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    • 제41권2호
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    • pp.67-74
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    • 2020
  • Brain tumors or gliomas are fatal cancer species with high recurrence rates due to their strong invasiveness. Therefore, the goal of surgery is complete tumor resection. However, the surgery is difficult to distinguish the border because tumors and blood vessels have the same color tone and shape. The fluorescein sodium is used as a fluorescence contrast agent for boundary separation. When the external light source is irradiated, yellow fluorescence is expressed in the tumor, which helps distinguish between blood vessels and tumor boundaries. But, the fluorescence expression of fluorescence sodium depends on the concentration of fluorescein sodium and such analytical data is insufficient. The unclear fluorescence can obscure the boundaries between blood vessels and tumors. In addition, reduce the efficiency of fluorescence sodium use. This paper proposes a protocol of concentration range for fluorescence expression conditions. Fluorescent expression was observed using a near-infrared (NIR) color camera with corresponding dilution using normal saline in 1 ml microtube. The flunoresence emission density range is 1.00 mM to 0.15 mM. The fluorescence emission begin to 1.00 mM and the 0.15 mM discolor. The discolor is difficult to fluorescence emission condition obserbation. Thus, the maximum density range of the bright fluoresecein is 0.15 mM to 0.30 mM. When the concentration range of fluorescein sodium is analyzed based on the gradient of fluorescence expression and the power measurement, the brightest fluorescence is expected to facilitate the complete resection of the tumor. For the concentration range protocol, setting concentration ranges and analyzing fluorescence expression image according to saturation and brightness to find optimal fluorescence concentration are important. Concentration range protocols for fluorescence expression conditions can be used to find optimal concentrations of substances whose expression pattern varies with concentration ranges. This study is expected to be helpful in the boundary classification and resection of brain tumors and glioma.

Calmodulin 단백질의 형태변화를 이용한 광섬유 형광센서에 의한 $Ca^{2+}$의 정량 (Determination of $Ca^{2+}$ by Fiber Optic Fluorosensor Based on the Conformational Change of the Protein Calmodulin)

  • 이창섭;양승태
    • 분석과학
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    • 제8권3호
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    • pp.221-227
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    • 1995
  • $Ca^{2+}$에 대하여 특이한 선택성을 보이는 광섬유형광센서에 대하여 연구하였다. 이 센서는 $Ca^{2+}$과 형광성 킬레이트를 형성하는 단백질 Calmodulin(CaM)을 사용하였으며, 두 갈래로 된 광섬유 다발의 끝면에 플루오르세인 이소티오시아네이트로써 형광 표지된 Calmodulin(FCaM)으로 만든 용액을 투석막 안에 넣어서 제작하였다. 이 센서의 감응 메카니즘은 FCaM이 $Ca^{2+}$과 결합하여 킬레이트를 형성할 때에 나타나는 형광 스펙트럼의 이동 현상을 바탕으로 한다. CaM은 $Ca^{2+}$과 결합할 때에 형태변화를 일으키며, 이로 인해 유발되는 FCaM의 형광세기 변화로써 농도를 결정하였다. 광전자증배관으로 형광의 세기를 측정하여 $Ca^{2+}$에 대한 검정곡선을 작성하였으며, 센서의 $Ca^{2+}$에 대한 검출한계와 $Mg^{2+}$, $Eu^{3+}$, $La^{3+}$들에 의한 방해효과, 감응 시간 및 수명을 조사하였다.

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Lipid-binding properties of TRIM72

  • Kim, Sung-Hyen;Seo, Jeong-Hwa;Ko, Young-Gyu;Huh, Young-Duk;Park, Heon-Yong
    • BMB Reports
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    • 제45권1호
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    • pp.26-31
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    • 2012
  • TRIM72 is known to play a critical role in skeletal muscle membrane repair. To better understand the molecular mechanisms of this protein, we carried out an in vitro binding study with TRIM72. Our study proved that TRIM72 binds various lipids with dissociation constants ($K_d$) ranging from 88.2 ${\pm}$ 9.9 nM to 550.5 ${\pm}$ 134.5 nM. In addition, the intrinsic fluorescence of TRIM72 exponentially decreased when the protein was diluted with stirring. The time-resolved fluorescence decay occurred in a concentration-independent manner. The fluorescence-decayed TRIM72 remained in its secondary structure, but its binding properties were significantly reduced. The dissociation constants ($K_d$) of fluorescence-decayed TRIM72 for palmitate and stearate were 159.1 ${\pm}$ 39.9 nM and 355.4 ${\pm}$ 106.0 nM, respectively. This study suggests that TRIM72 can be dynamically converted by various stimuli. The results of this study also provide insight into the role of TRIM72 in the repair of sarcolemma damage.

Highly Fluorescing Solid DNA-Cationic Polyelectrolyte Complexes Prepared from a Natural DNA and a Poly(fluorenevinylene-alt-phenylene) Bearing Quaternary Ammonium Pendants

  • Yu, Young-Jun;Kwon, Young-Wan;Kim, Kyu-Nam;Do, Eui-Doo;Choi, Dong-Hoon;Jin, Jung-Il;Shin, Hee-Won;Kim, Yong-Rok;Kang, Ik-Joong;Mikroyannidis, John A.
    • Macromolecular Research
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    • 제17권4호
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    • pp.245-249
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    • 2009
  • A fluorescing, copolymer(Q)-bearing, quaternary ammonium pendant was mixed with excess natural salmon sperm DNA with a molecular weight of $1.3{\times}10^6$(2,000 base pairs) to afford highly fluorescing, complex mixtures. The fluorescence life-time of the polymer Q was greatly increased when mixed with DNA: for the mixture of Q:DNA=1:750 the fast and slow decay lifetimes increased from ca. 10 to 100 ps and from 20 ps to ca. 1 ns, respectively. The enhanced fluorescence of the mixtures was ascribed to efficient compartmentalization and reduced conformational relaxation of the polymer Q by complexation with excess DNA.

Design of Home or Hetero Polynuclear Metal Dendritic Molecule for PDD or PDT

  • Choi, Chang-Shik
    • 한국정보통신학회:학술대회논문집
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    • 한국정보통신학회 2021년도 추계학술대회
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    • pp.646-648
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    • 2021
  • The syntheses and properties of polynuclear metal complexes have been reported to develop the easy syntheses and noble photo-characteristics of those complexes for photodynamic diagnosis (PDD) or photodynamic therapy (PDT). We have been focused on the design and synthesis of polynuclear lanthanide dendritic molecule due to long life time of fluorescence. Therefore, we will be presented on the design of home (Eu or Gd) or hetero (Tb or Lu) polynuclear lanthanide dendritic molecule.

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Photosynthetic Response and Protective Regulation To Ultraviolet-B Radiation In Green Pepper (Capsicum annuum L.)Leaves

  • Kim, Dae-Whan;Jun, Sung-Soo;Hong, Young-Nam
    • Journal of Photoscience
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    • 제8권1호
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    • pp.1-7
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    • 2001
  • The deteriorative effect of ultraviolet-B(UV-B) radiation on photosynthesis was assessed by the simultaneous measurement of O$_2$ evolution and chlorophyll(Chl) fluorescence in green pepper. UV-B was given at the intensity of 1 W$.$m$\^$-2/, a dosage often encountered in urban area of Seoul in Korea, to detached leaves. Both Pmax and quantum yield of O$_2$ evolution was rapidly decreased, in a parallel phase, with increasing time of UV-B treatment. Chl fluorescence parameters were also significantly affected. Fo was increased while both Fm and Fv were decreased. Photochemical efficiency of PSII(Fv/Fm) was also declined, although to a lesser extent than Pmax. Both qP and NPQ were decreased similarly with increasing time of UV-B treatment. However, PS I remained stable. The addition of lincomycin prior to UV-B treatment accelerated the decline in Fv/Fm to some extent, suggesting that D1 protein turnover may play a role in overcoming the harmful effect of UV-B. The amount of photosynthetic pigments was less affected than photosynthetic response in showing decline in Chl a and carotenoids after 24 h-treatment. Presumptive flavonoid contents, measured by changes in absorbance at 270 nm , 300 nm and 330nm, were all increased by roughly 50% after 8 h-treatment. Among antioxidant enzymes, activities of catalase and peroxidase were steadily increased until 12h of UV-B treatment whereas ascorbate perxidase, dehydroascorvate reductase and glutathione reductase did not show any significant change. The results indicate that deteriorative effect of UV-B on photosynthesis precedes the protection exerted by pigment synthesis and antioxidant enzymes.

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