• 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.

• Archives of Pharmacal Research
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• v.15 no.3
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• pp.196-203
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• 1992

Intramolecular excimer formation of 1, 3-di(1-pyrenyl)propane (Py-3-Py) and fluorescence polarization of 1, 6-diphenyl-1, 3, 5-hexatriene (DPH) were used to investigate the effects of barbiturates on the fluidity of model membranes of phosphatidycholine (SPMVPC), phosphatidylserine (SPMVPS), and phosphatidylinositol (SPMVPI) fractions of synaptosomal plasma membrane vesicles (SPMV) isolated from bovine cerebral cortex. In a dose-dependent manner, barbiturates decreased the excimer to monomer fluorescence intensity ratio (I'/I) of Py-3-Py and increased the anisotropy(r), rotational relaxation time (P), limiting anisotropy $(r_infty)$, and order parameter (S) of DPH in SPMVPC, SPMVPS and SPMVPI. This indicates that barbiturates decreased both the lateral and rotational diffusion of the probes in SPMVPC, SPMVPS and SPMVPI. The relative potencies of barbiturates in ordering the membranes were in the order: pentobarbital > hexobarbital > amobarbital > phenobarbital. This order correlates well with the anesthetic potencies of barbiturates and the potencies for enhancement of $\gamma$-aminobutyric acid-stimulated chloride uptake. Thus, it is strongly suggested that a close relationship might exist between the membrane ordering effects of barbiturates and the chloride fluxes across SPMV.

• Bulletin of the Korean Chemical Society
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• v.33 no.5
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• pp.1608-1612
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• 2012

Singlet oxygen ($^1O_2$) is an important species for oxidation in biological processes. $^1O_2$ is implicated in the genotoxic effect, and plays an important role in the cell-signaling cascade and in the induction of gene expression. However, the rapid detection of $^1O_2$ in biological environments with sufficient specificity and sensitivity is hampered by its extremely low emission probability. Here, a layer-by-layer (LbL) film of CdTe quantum dots (QDs), polymers, and ascorbate have been designed as a rapid, highly selective, and sensitive fluorescence probe for $^1O_2$ detection. Upon reaction with $^1O_2$, the probe exhibits a strong photoluminescence (PL) response even at trace levels. This remarkable PL change should enable the probe to be used for $^1O_2$ detection in many chemical and biological systems and as an environmental sensor.

• Journal of Periodontal and Implant Science
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• v.38 no.4
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• pp.691-698
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• 2008

Purpose: Periodontal pathogens can invade the host tissue. Morphologic studies have revealed bacteria within the pocket epithelium, gingival connective tissues, alveolar bone, and oral epithelium. The objective of this study was to visualize and evaluate presence of Porphyromonas gingivalis and Tannerella forsythia in crevicular epithelial cells of periodontally healthy subjects and chronic periodontitis patients. Materials and Methods: A total of 666 crevicular epithelial cells in the samples obtained from 27 chronic periodontitis patients and 9 healthy volunteers were examined. Specific probes for P. gingivalis and T. forsythia and a universal probe for detection of all eubacteria targeting 168 rRNA for fluorescence in situ hybridization was used in conjunction with confocal laser scanning microscopy. Results: 98.99% of sulcular epithelial cells from healthy volunteers and 84.40% of pocket epithelial cells from periodontitis patients were found to harbor bacteria. P. gingivalis and T. forsythia were discovered more often in crevicular epithelial cells from periodontitis patients. Conclusion: P. gingivalis and T. forsythia can invade crevicular epithelial cells and intracellular bacteria may act as a source of bacteria for persistent infection.

• Korean Journal of Breeding Science
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• v.43 no.4
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• pp.252-259
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• 2011

Conventional staining and fluorescence in situ hybridization (FISH) karyotypes of the non-genetically modified (GM) parental rice line, 'Nakdong' (Oryza sativa L. japonica), and its four GM rice lines, LS28 (event LS30-32-20-1), Cry1Ac1 (event C7-1-9-1), and LS28 ${\times}$ Cry1Ac1 (events L/C1-1-3-1 and L/C1-3-1-1) were analyzed using 5S and 45S rDNAs as probes. Both parental and transgenic lines were diploids (2n=24) with one satellite chromosome pair. The lengths of the prometaphase chromosomes ranged from 1.50 to $6.30{\mu}m$. Four submetacentric and eight metacentric pairs comprised the karyotype of 'Nakdong' and its four GM lines. One pair of 5S rDNA signals was detected near the centromeric region of chromosome g in both the parental and transgenic lines. The 45S rDNA signals were detected on the secondary constrictions of the satellite chromosome pair in both the parental and transgenic lines. There was no significant difference in chromosome size, length, and composition between 'Nakdong' and its four GM lines. This research was conducted as a preliminary study for chromosomal detection of transgenes in GM rice lines and would be useful for their breeding programs.

• Annals of Clinical Microbiology
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• v.18 no.2
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• pp.37-43
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• 2015

Background: Tuberculosis is globally the most important cause of death from single pathogen. Rapid and accurate identification of mycobacteria is essential for the control of tuberculosis. We evaluated a fluorescence in situ hybridization (FISH) method using peptide nucleic acid (PNA) probes for the differentiation of Mycobacterium tuberculosis complex (MTB) and nontuberculous mycobacteria (NTM) in direct smears of sputum specimens. Methods: The cross-reactivity of MTB- and NTM-specific PNA probes was examined with reference strains of M. tuberculosis ATCC 13950, Mycobacterium kansasii ATCC 12479, Mycobacterium fortuitum ATCC 6841, several clinical isolates of mycobacteria (Mycobacterium abscessus, Mycobacterium avium, Mycobacterium intracellulare, Mycobacterium gordonae and Mycobacterium chelonae), and 11 frequently isolated respiratory bacterial species other than mycobacteria. A series of 128 sputa (89 MTB culture positive, 29 NTM culture positive, and 10 under treatment culture negative) with grades of trace to 4+ were used to evaluate the performance of the method. Results: The MTB- and NTM-specific PNA probes showed specific reactions with the reference strains of MTB and M. kansasii and clinical isolates of mycobacteria except M. fortuitum ATCC 6841, and no cross-reactivity with other tested bacteria. The PNA probe-based FISH assay for detection of MTB had a sensitivity and specificity of 100%, respectively. The sensitivity and specificity of the NTM-specific PNA probe was 100%. The smear grades of the PNA FISH test were same as with those of the fluorescence AFB stain in 2+ or higher grade. Conclusion: Detection and differentiation based on PNA FISH is sensitive and accurate for detecting mycobacteria and for differentiating MTB from NTM in clinical sputum smears.

• Journal of Pharmaceutical Investigation
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• v.21 no.3
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• pp.133-141
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• 1991

The fluorescence characteristics of l-anilinonaphthalene-8-sulfonate (ANS) and 2-p-toluidinylnaphthalene-6-sulfonate (TNS) made the dyes useful probes for the determination of the polarity at the binding sites of several water-soluble polyparacyclophanes. Polyparacyclophanes used were 1,6,20,25-tetraaza[ 6.1.6.1]paracyclophane (CPM 44), 1,7,21,27 -tetraaza[7.1.7.1]paracyclophane (CPM 55). 1,7,21,27 -tetraaza-14,34-dioxa[7.1.7.1]paracyclophane (CPE 55) and 1,8,22,29-tetraaza-15,36-dioxa[8.1.8.1] paracyclophane (CPE 66). The fluorescence quantum yield, emission maximum, and half bandwidth of ANS or TNS obtained in a variety of solvent systems were plotted as a function of four kinds of empirical solvent polarity scales such as dielectric constant (D), (D-l)/(2D+1). Y and Z values. It was found that the Z-value-emission maximum $(\overline}V_F,\;cm^{-1})$ profile showed the most reliable linearity. ANS and TNS interacted with CPM 44, CPM 55, CPE 55. CPE 66. ${\alpha}-cyclodextrin$ (CyD) and ${\beta}-CyD$ in the aqueous solution, and from the emission maxima the polarities (Z-value) of their binding sites were calculated to be 92.65, 87.50, 93.35, 84.52, 94.36, and 90.48 for ANS, respectively. and 91.07, 89.68, 85.44, 86.74 and 87.6 for TNS except for ${\alpha}-CyD$, respectively.

• Clinical and Experimental Reproductive Medicine
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• v.24 no.3
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• pp.369-375
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• 1997

Studies were conducted to determine the efficiency of decondensation protocols. Sperm obtained from seven normal donors was immediately washed after liquefaction and then decondensed using the method of West et al. (1989) and my original protocol. My optimized protocol entailed mixing 1 ml aliquots of semen with 4 ml phosphate buffered saline (PBS). Following centrifugation, pellets were resuspended in 1 ml PBS containing 6 mM EDTA. After centrifugation, pellets were resuspended in 1 ml PBS containing 2 mM dithiothreitol at $37^{\circ}C$ for 45 min. Following mixing with 2 ml PBS and centrifugation, pellets were resuspended by vortexing. While vortexing, 5 ml of fixative were gently added. Slide preparation was accomplished using the smear method and it was stored at $4^{\circ}C$. When comparing these protocols, the degree of sperm decondensation and head swelling was monitored by measuring nuclear length, area, perimeter, and degree of roundness using FISH analysis software. Apparent copy number for chromosome 1 and, separately, for the sex chromosomes was determined by FISH using satellite DNA probes for loci DIZ1, DXZ1 and DYZ3. Sperm treated by my decondensation protocol showed significant increases (p<0.05) in length, area, perimeter, and degree of roundness. There was a significant decrease (p<0.05) in the frequency of nuclei displaying no signal but no change in the frequency of nuclei with two signals in samples decondensed by my protocol. My data suggested that decondensation using my original protocol may lower the frequency of cells with spurious "nullisomy" due to hybridization failure without inducing spurious "disomy" resulting from increased distances between split signals.

• Biomaterials Research
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• v.22 no.4
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• pp.352-359
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• 2018

Background: Black phosphorus (BP) has emerged as a novel class of nanomaterials owing to its unique optical and electronic properties. BP, a two-dimensional (2D) nanomaterial, is a structure where phosphorenes are stacked together in layers by van der Waals interactions. However, although BP nanodots have many advantages, their biosafety and biological effect have not yet been elucidated as compared to the other nanomaterials. Therefore, it is particularly important to assess the cytotoxicity of BP nanodots for exploring their potentials as novel biomaterials. Methods: BP nanodots were prepared by exfoliation with a modified ultrasonication-assisted solution method. The physicochemical properties of BP nanodots were characterized by transmission electron microscopy, dynamic light scattering, Raman spectroscopy, and X-ray diffractometry. In addition, the cytotoxicity of BP nanodots against C2C12 myoblasts was evaluated. Moreover, their cell imaging potential was investigated. Results: Herein, we concentrated on evaluating the cytotoxicity of BP nanodots and investigating their cell imaging potential. It was revealed that the BP nanodots were cytocompatible at a low concentration, although the cell viability was decreased with increasing BP nanodot concentration. Furthermore, our results demonstrated that the cells took up the BP nanodots, and the BP nanodots exhibited green fluorescence. Conclusions: In conclusion, our findings suggest that the BP nanodots have suitable biocompatibility, and are promising candidates as fluorescence probes for biomedical imaging applications.

• 한국생물공학회:학술대회논문집
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• 2000.11a
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• pp.508-511
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• 2000

In situ hybridization with fluorescent oligonucleotides(FISH) was used to detect and localize microorganisms in the granules of lab-scale upflow anaerobic sludge blanket(UASB) reactors. An UASB reactor was seeded with mesophilically-grown($35^{\circ}\;C$) granular sludge, and thermophilically($55^{\circ}\;C$) operated by feeding with a synthetic wastewater. Sections of the granules were hybridized with 16S rRNA-targeted oligonucleotide probes for Eubacteria, Archaeabacteria, and specific phylogenetic groups of methanogens. FISH clearly showed the layed structure of thermophilic granules, which was consisted of outer bacterial cells and inner archaeal cells. Methanoseata-, Methanosarcina-like cells were also found to be localized inside the granules. These results demonstrated FISH was useful in studying the spatial organizations of methanogens and in situ morphologies and metabolic functions in thermophilic granular sludges.