• Title/Summary/Keyword: Fluorescent probe

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Chromosome Aberrations in Porcine Embryo Produced by Nuclear Transfer with Somatic Cell

  • K. S. Chung;Ko, S. A;S. J. Song;J. T. Do;Park, Y. S.;Lee, H. T.
    • Korean Journal of Animal Reproduction
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    • v.26 no.4
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    • pp.385-394
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    • 2002
  • This study was constructed the correlations of the embryonic developmental rates and the frequency of chromosome aberration using ear-skin-fibroblast cell in nuclear transfer (NT) derived embryos. Karyoplast-oocyte complexes were fused and activated simultaneously, then cultured for seven days to assess development. The developmental rates of NT and in vitro fertilization (IVF) embryos were 55.4% vs 63.5%, 31.7% vs 33% and 13.4% vs 16.8% in 2 cell, 8 cell and blastocyst, respectively. Firstly, the frequency of chromosome aberrations were evaluated using fluorescent in situ hybridization (FISH) technique with porcine chromosome 1 submetacentric specific probe. Chromosome aberration was detected at day 3 on the embryo culture, the percentages of chromosomal aneuploidy in NT and IVF embryos at 4-cell stage were 40%, 31.3%, respectively. Secondly, embryonic fragmentation was evaluated at 4-cell stage embryo. Frequency of embryonic fragmentations was in 51.3% of NT, 61.3% of IVF, 28.9% of parthenogenetic activation at 4-cell stage. The proportion of fragmentation in NT embryos was higher than activation embryos. This result indicates that chromosomal abnormalities and embryonic fragments are associated with low developmental rate in porcine NT embryo. It is also suggest that abnormal porcine embryos produced by NT related with lower implantation rate, increased abortion rate and production of abnormal fetuses.

Effects of Local Anesthetics on Rotational Mobility of n-(9-Anthroyloxy)stearic Acid in Neuronal Membranes

  • Jang, Hye-Ock;Lee, Chang;Choi, Min-Gak;Shin, Sang-Hun;Chung, In-Kyo;Yun, Il
    • The Korean Journal of Physiology and Pharmacology
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    • v.7 no.3
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    • pp.119-124
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    • 2003
  • To elucidate the molecular mechanism of pharmacological action of local anesthetics, we studied membrane actions of tetracaine, bupivacaine, lidocaine, prilocaine and procaine. Fluorescence polarization of n-(9-anthroyloxy)stearic acid (n-AS) was used to examine the effects of these local anesthetics on differential rotational mobility of different positions of the number of synaptosomal plasma membrane vesicle (SPMV) phospholipid carbon atoms. The four membrane components differed with respect to 3, 6, 9 and 16-(9-anthroyloxy)stearic acid (3-AS, 6-AS, 9-AS and 16-AP) probes, indicating that differences in the membrane fluidity might be present. Degrees of the rotational mobility of 3-AS, 6-AS, 9-AS and 16-AP were different depending on depth of hydrocarbon interior. In a dose-dependentmanner, tetracaine, bupivacaine, lidocaine, prilocaine and procaine decreased anisotropy of 3-AS, 6-AS, 9-AS and 16-AP in the hydrocarbon interior of the SPMV. These results indicate that local anesthetics have significant disordering effects on hydrocarbon interior of the SPMV, thus affecting the transport of $Na^+$ and $K^+$ in nerve membranes and leading to anesthetic action.

Characterization and bacterial anti-adherent effect on modified PMMA denture acrylic resin containing platinum nanoparticles

  • Nam, Ki-Young
    • The Journal of Advanced Prosthodontics
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    • v.6 no.3
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    • pp.207-214
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    • 2014
  • PURPOSE. This study characterized the synthesis of a modified PMMA (Polymethyl methacrylate) denture acrylic loading platinum nanoparticles (PtN) and assessed its bacterial inhibitory efficacy to produce novel antimicrobial denture base material. MATERIALS AND METHODS. Polymerized PMMA denture acrylic disc ($20mm{\times}2mm$) specimens containing 0 (control), 10, 50, 100 and 200 mg/L of PtN were fabricated respectively. The obtained platinum-PMMA nanocomposite (PtNC) was characterized by TEM (transmission electron microscopy), SEM/EDX (scanning electron microscope/energy dispersive X-ray spectroscopy), thermogravimetric and atomic absorption spectrophotometer analysis. In antimicrobial assay, specimens were placed on the cell culture plate, and $100{\mu}L$ of microbial suspensions of S. mutans (Streptococcus mutans) and S. sobrinus (Streptococcus sobrinus) were inoculated then incubated at $37^{\circ}C$ for 24 hours. The bacterial attachment was tested by FACS (fluorescence-activated cell sorting) analysis after staining with fluorescent probe. RESULTS. PtN were successfully loaded and uniformly immobilized into PMMA denture acrylic with a proper thermal stability and similar surface morphology as compared to control. PtNC expressed significant bacterial anti-adherent effect rather than bactericidal effect above 50 mg/L PtN loaded when compared to pristine PMMA (P=.01) with no or extremely small amounts of Pt ion eluted. CONCLUSION. This is the first report on the synthesis and its antibacterial activity of Pt-PMMA nanocomposite. PMMA denture acrylic loading PtN could be a possible intrinsic antimicrobial denture material with proper mechanical characteristics, meeting those specified for denture bases. For clinical application, future studies including biocompatibility, color stability and warranting the long-term effect were still required.

Physiological Activities of Peel of Jeju-indigenous Citrus sunki Hort. Tanaka (제주자생 진귤(Citrus sunki Hort. Tanaka) 과피의 생리활성)

  • Kang, Shin-Hae;Lee, Young-Jae;Lee, Chang-Hong;Kim, Se-Jae;Lee, Dae-Ho;Lee, Young-Ki;Park, Deok-Bae
    • Korean Journal of Food Science and Technology
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    • v.37 no.6
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    • pp.983-988
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    • 2005
  • Effects of Citrus sunki peel and its fermented product extracts on physiological and functional activities of cellular systems were investigated. Ethanol extract of Citrus sunki peel showed potent ROS-scavenging activity using 2',7'-Dichlorofluorescin diacetate as a fluorescent ROS probe in HepG2 cells. Fermented product of C. sunki peel extract markedly suppressed nitric oxide production in lipopolysaccharide (LPS)-activated RAW264.7 murine macrophage cells. Treatment with fermented product of C. sunki peel extract decreased intracellular protein levels of inducible nitric oxide synthase and cyclooxygenase II stimulated by LPS. High doses of fermented product lend to apoptotic cell death in CHO-IR cells.

Gene Expression Profile of Zinc-Deficient, Homocysteine-Treated Endothelial Cells

  • Kwun, In-Sook;Beattie, John H.
    • Preventive Nutrition and Food Science
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    • v.8 no.4
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    • pp.390-394
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    • 2003
  • In the post-genome period, the technique for identifying gene expression has been progressed to high throughput screening. In the field of molecular nutrition, the use of screening techniques to clarify molecular function of specific nutrients would be very advantageous. In this study, we have evaluated Zn-regulated gene expression in Zn-deficient, homocystein-treated EA.hy926 cells, using cDNA microarray, which can be used to screen the expression of many genes simultaneously. The information obtained can be used for preliminary assessment of molecular and signaling events modulated by Zn under pro-atherogenic conditions. EA.hy926 cells derived from human umbilical vein endothelial cells were cultured in Zn-adequate (control, 15 $\mu$M Zn) or Zn-deficient (experimental, 0 $\mu$M Zn) Dulbecco's MEM media under high homocysteine level (100 $\mu$M) for 3 days of post-confluency. Cells were harvested and RNA was extracted. Total RNA was reverse-transcribed and the synthesized cDNA was labeled with Cy3 or Cy5. Fluorescent labeled cDNA probe was applied to microarray slides for hybridization, and the slide was then scanned using a fluorescence scanner. The expression of seven genes was found to be significantly decreased, and one significantly increased, in response to treatment of EA.hy926 cells with Zn-deficient medium, compared with Zn-supplemented medium. The upregulated genes were oncogenes and tumor suppressor genes, cell cycle-related genes and transporter genes. The down-regulated gene was RelB, a component of the NF-kappaB complex of transcription factors. The results of this study imply the effectiveness of cDNA microarray for expression profiling of a singly nutrient deficiency, namely Zn. Furthur study, using tailored-cDNA array and vascular endothelial cell lines, would be beneficial to clarify the molecular function of Zn in atherosclerosis, more in detail.

Cytotaxonomical Study of the Chenopodium album and its Related Species in Korea (한국산 흰명아주와 근연종의 세포분류학적 연구)

  • Chung, Youngjae;l Kim, Muyeol;Lee, Byongsoon
    • Korean Journal of Plant Taxonomy
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    • v.41 no.4
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    • pp.324-328
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    • 2011
  • The purpose of this study was to analyze the interspecific relationships of Chenopodium album and its related taxa collected in Korea. The 18S-26S ribosomal DNA (45S rDNA) loci were detected directly on mitotic chromosomes by fluorescence in situ hybridization (FISH) and the chromosome numbers were examined using aceto-orcein methods. The chromosomal numbers of Chenopodium album var. album and C. album var. centrorubrum were 2n = 6x = 54, whereas for C. album var. stenophyllum, this number was 2n = 4x = 36. The basic chromosome number was x = 9. The biotin labeled 18S-26S rDNA probe exhibited eight yellow fluorescent signals on the metaphase chromosome of C. album var. album and var. centrorubrum respectively, while two yellow signals of C. album var. stenophyllum were noted. All of the signals on the chromosomes were located at the terminal regions. The chromosome number and FISH findings suggest that C. album var. centrorubrum is merged into var. album and that it is clearly distinguished from C. album var. stenophyllum.

DNA Dynamics: a Fluorescence Resonance Energy Transfer Study Using a Long-Lifetime Metal-Ligand Complex

  • Kang, Jung-Sook;Lakowicz, Joseph-R.;Piszczek, Grzegorz
    • Archives of Pharmacal Research
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    • v.25 no.2
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    • pp.143-150
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    • 2002
  • Fluorescent probes bound to DNA typically display nanosecond decay times and reveal only nanosecond motions. We extend the time range of measurable DNA dynamics using $[Ru(pby)_2(dppz)]^{2+}$ (bpy=2.2'-bipyridine, dppz=dipyrido[3,2-a2',3'-c]phenazine) (RuBD) which displays a mean lifetime near 90 ns. To test the usefulness of RuBD as a probe for diffusive processes in calf thymus DNA, we compared the efficiencies of fluorescence resonance energy transfer (FRET) using three donors which display lifetimes near 5 ns for acridine orange (AO), 22 ns for ethidum bromide (EB) and 92 ns for RuBD, with nile blue (NB) as the acceptor. The F rster distances for AO-NB, EB-NB and RuBD-NB donor-acceptor pairs were 42.3, 52.3, and $30.6{\;}{\AA}$, respectively. All three donors showed dramatic decreases in fluorescence intensities and more rapid intensity decays with increasing NB concentrations. The intensity decays of AO and EB in the presence of varying concentrations of NB were satisfactorily described by the one-dimensional FRET model without diffusion (Blumen and Manz, 1979). In the case of the long-lifetime donor RuBD, the experimental phase and modulation somewhat deviated from the recovered values computed from this model. The recovered NB concentrations and FRET efficiencies from the model were slightly larger than the expected values, however, the recovered and expected values did not show a significant difference. Thus, it is suggested that the lifetime of RuBD is too short to measure diffusive processes in calf thymus DNA.

Molecular Analysis of the Y Chromosome in a 46,XY Female Phenotype

  • Kim, Jin-Woo;Kim, Tae-Jin;Park, So-Yeon;Nam, Sung-A;Jun, Jong-Young
    • Journal of Genetic Medicine
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    • v.3 no.1
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    • pp.5-10
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    • 1999
  • This is a case report of 46,XY female phenotype (46,XY karyotype, no pubic hair, blind vagina and absence of uterus)in an 18-year-old patient. To confirm whether a Y chromosome has a structural abnormality, fluorescent in situ hybridization (FISH) with the chromosome X/Y cocktail probe was simultaneously performed, and the six loci [PABY, RPS4Y(sy16, sy17), ZFY, DYS14] on the short arm, one locus (DYZ3) on the centromere and one locus (DYZ1) on the long arm were amplified by polymerase chain reaction (PCR). The probes used FISH hybridized to centromere of the X chromosome and heterochromatin region (Yq12) of the Y chromosome, and all PCR related Y chromosome showed positive band like normal male. From the results obtained, it seemed that the Y chromosome from the 46,XY female was structurely normal. Especially, the SRY gene has been equated with the mammalian testis-determining factor, and absence or point mutation in the SRY gene causes XY female. To detect the point mutations of SRY sequences, single-strand conformation polymorphism (SSCP) assay was used. Our results confirm that this patient has no mutation in the SRY gene on the Y chromosome.

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A Themotropic Behavior of Egg PC Liposome Containing the Very Long Chain Fatty Acyl Component,${\alpha},{\omega}$-13,16-Dimethyloctacosanedioate Dimethyl Ester(DME C30) Isolated from The Thermophilic Anaerobic Bacteria, Thermoanaerobacter ethanolicus

  • Kim, Hyeon Myeong;Gang, Se Byeong;Jeong, Seung Ho
    • Bulletin of the Korean Chemical Society
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    • v.22 no.9
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    • pp.979-983
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    • 2001
  • Thermoanaerobacter ethanolicus is a strictly anaerobic and thermophilic bacterium whose optimum temperature ranges over $65-68^{\circ}C.$ T. ethanolicus was known to contain a bipolar very long chain fatty acyl component such as $\alpha$, $\omega-1316-dimethyloctacosanedioate$, as one of the major membrane components. However, exact physiological role of this unusual component in the membrane remains unknown. Such a very long chain fatty acyl component, $\alpha$, ${\omega}-1316-dimethyloctacosanedioate$, dimethyl ester (DME C30), was isolated, and purified from the membrane of T. ethanolicus. As a function of added concentrations of the $\alpha$, $\omega-1316-dimethyloctacosanedioate$, dimethyl ester (DME C30) or cholesterol into the standard liposomes, the acyl chain ordering effect was investigated by the steady-state anisotropy with 1,6-diphenyl-1,3,5-hexatriene (DPH) as a fluorescent probe. Acyl chain order parameter (S) of vesicles containing DME C30 is higher comparing with phosphatidylcholine (PC) only vesicles. This result was discussed thermodynamically with the aid of the simulated annealing molecular dynamics simulations. Through the investigation of all the possible conformational changes of DME C30 or cholesterol, we showed that DME C30 is very flexible and its conformation is variable depending on the temperature comparing with cholesterol, which is rigid and restricted at overall temperature. We propose that the conformational change of DME C30, not the configurational change, may be involved in the regulation of the membrane fluidity against the changes of external temperature.

Green Synthesis of Multifunctional Carbon Nanodots and Their Applications as a Smart Nanothermometer and Cr(VI) Ions Sensor

  • Li, Lu;Shao, Congying;Wu, Qian;Wang, Yunjian;Liu, Mingzhu
    • Nano
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    • v.13 no.12
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    • pp.1850147.1-1850147.14
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    • 2018
  • In this work, water-soluble and blue-emitting carbon nanodots (CDs) were synthesized from apple peels for the first time via one-step hydrothermal method. The synthetic route is facile, green, economical and viable. The as-prepared CDs were characterized thoroughly by transmission electron microscopy (TEM), X-ray diffraction (XRD), Raman, Fourier transform infrared (FT-IR), X-ray photoelectron (XPS), fluorescence and UV-Vis absorption spectroscopy in terms of their morphology, surface functional groups and optical properties. The results show that these CDs possessed ultrasmall size, good dispersivity, and high tolerance to pH, ionic strength and continuous UV irradiation. Significantly, the CDs had fast and reversible response towards temperature, and the accurate linear relationship between fluorescence intensity and temperature was used to design a novel nanothermometer in a broad temperature range from 5 to $65^{\circ}C$ facilely. In addition, the fluorescence intensity of CDs was observed to be quenched immediately by Cr(VI) ions based on the inner filter effect. A low-cost Cr(VI) ions sensor was proposed employing CDs as fluorescent probe, and it displayed a wide linear range from 0.5 to $200{\mu}M$ with a detection limit of $0.73{\mu}M$. The practicability of the developed Cr(VI) sensor for real water sample assay was also validated with satisfactory recoveries.