• Title/Summary/Keyword: multi confocal system

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Multi-dimensional analyses of plant chromosomes and genomes.

  • Fukui, Kiichi;Ohmido, Nobuko;Wako, Toshiyuki
    • Proceedings of the Botanical Society of Korea Conference
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    • 1998.07a
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    • pp.61-70
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    • 1998
  • Genome and chromosome analyses in plants using fluorescence in situ hybridization (FISH) and immuno-staining (IMS) methods are reviewed by presenting the recent results obtained by the Chromosome Link, a group of chromosome and genome researchers. FISH is now effective to detect unique nucleotide sequences with 153 bp on the extended DNA fibers. Genomic in situ hybridization (GISH) also allows painting plant chromosomes of different genomes. GISH is quite effective to detect the genomic differentiation in the individual chromosomes within a nucleus. Three dimensional (3D) analyses are now available by confocal microscopy and a deconvolution system. These techniques are invaluable to visualize both the structural and functional dynamics within a nucleus. 3D-FISH revealed the spatial differentiation of different genomees within a nucleus. 3D-FISH also proved structural partition of centromeric and telomeric domains within a barely nucleus. The dynamic acetylation of histone H4 at the specific regions of a genome during a cell cycle is also analyzed using 3D-IMS. It is anticipated that these methods will provide us powerful tools to understand the structural and functional significance of plant chromosomes and genomes.

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CONFOCAL LASER SCANNING MICROSCOPIC MORPHOLOGY OF DENTIN-RESIN INTERFACE AND ITS RELATIONSHIP WITH SHEAR BOND STRENGTH (상아질-레진 계면의 공초점 현미경적 형태 및 전단결합강도와의 관계)

  • Choi, Nak-Won;Cho, Byeong-Hoon;Son, Ho-Hyun
    • Restorative Dentistry and Endodontics
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    • v.24 no.2
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    • pp.310-321
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    • 1999
  • In this in vitro study, confocal laser scanning microscopic morphology of dentin-resin interface and its relationship to shear bond strength were investigated after the exposed dentin surfaces were treated with 3 different kinds of dentin adhesive systems[three-step; Scotchbond Multi-Purpose Plus(SMPP), self-priming bonding resin; Single Bond(SB), self-etching primer; Clearfil Liner Bond 2(LB2)]. 52 extracted human molar teeth without caries and/or restorations. The experimental teeth were randomly divided into three groups of seventeen teeth each. In five teeth of each group, class V cavities(depth: 1.5mm) with 900 cavosurface angles were prepared at the cementoenamel junction on buccal and lingual surfaces. Bonding resins of each dentin adhesive system were mixed with rhodamine B. Primer of SMPP was mixed with fluorescein. In group 1. the exposed dentin was conditioned with etchant, applied with above primer and bonding resin of SMPP. In group 2, with etchant and self-priming bonding agent of SB. In group 3, with self-etching primer and bonding agent of LB2. After treatment with dentin adhesive systems, composite resin were applied and photocured. The experimental teeth were cut longitudinally through the center line of restoration and grounded so that about $90{\mu}m$-thick wafers of buccolingually orientated dentin were obtained. And, $70{\sim}80{\mu}m$-thick wafers sectioned horizontally, thus presenting a dentinal tubules at 900 to the cut surface of a remaining tooth, were obtained. Primer of SMPP mixed with rhodamine B was applied to these wafers. Confocal laser scanning microscopic investigations of these wafers were done within of 24 hours after treatment. To measure shear bond strength, the remaining twelve teeth of each group were grounded horizontally below the dentinoenamel junction, so that no enamel remained. After applying dentin adhesive systems on the dentin surface, composite was applied in the shape of cylinder. The cylinder was 5mm in diameter, and 2mm in thickness. Shear bond strength was measured using Instron with a cross-head speed of 0.5mm/min. It was concluded as follows ; 1. Hybrid layer of SMPP(mean: $4.56{\mu}m$) was thicker than that of any other groups. This value was not statistically significant thicker than that of SB(mean: $3.41{\mu}m$, p>0.05), and significant thicker than that of LB2(mean: $1.56{\mu}m$, p<0.05). There was a statistical difference between SB and LB2(p<0.05). 2. Although there were variations in the length of resin tag even in a sample, and in a group, most samples in SMPP and SB showed resin tags extending above $20{\mu}m$. But samples in LB2 showed resin tags of $10{\mu}m$ at best. 3. Besides primer's infiltration into demineralized peritubular dentin and dentinal tubules, fluorophore of primer was detected in the lateral branches of dentinal tubules. 4. All groups demonstrated statistically significant differences from one another(p<0.05), with shear bond strengths given in descending order as follows: SMPP(18.3MPa), SB(16.0MPa) and LB2(12.4MPa). 5. LB2 having thinnest hybrid layer($1.56{\mu}m$) showed the lowest shear bond strength(12.4MPa).

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A Study on the Stabilization of the Papain Enzyme in the Moderately Concentrated Anionic Surfactant System (음이온 계면활성제에서 파파인 효소의 안정도에 관한 연구)

  • Kim, Ji-Yeong;Kim, Jin-Woo;Kim, Yong-Jin;Lee, Jae-Wook;Lee, Hae-Kwang;Kang, Hak-Hee
    • Journal of the Society of Cosmetic Scientists of Korea
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    • v.33 no.2
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    • pp.93-97
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    • 2007
  • Even in the moderately concentrated anionic surfactant system, some special encapsulation method can shield the papain enzyme from proteolytic attacks. The stabilization of enzyme has been a major issue for successful therapies. In this study, we first stabilized an enzyme, papain in the microcapsules by using polyols, polyethyleneglycol (PEG), poly-propyleneglycol (PPG), and PEG-PPG-PEG block copolymer. In the analysis of EDS and CLSM, it was demonstrated that polyols are effectively located in the interface of papain and polymer. Polyols located in the interface had an ability to buffer the external triggers by hydrophobic partitioning, preventing consequently the catalytic activity of papain in the micro-capsules. Second. we introduced multi-layer capsulation methods containing ion complex. Such a moderately concentrated anionic surfactant system as wash-off cleansers, surfactants and waters can cause instability of entrapped enzymes. Surfactants and water in our final products swell the surface of enzyme capsules and penetrate into the core so easily that we can not achieve the effect of enzyme, papain. In this case, the ion complex multi-layer capsule composed of sodium lauroyl sarcosinate and polyquaternium-6 could effectively prevent water from penetration into the core enzyme, followed by in vivo test, and evaluate the stratum corneum (SC) turn-over speed.

Fabrication of Fluorescent Labeled Bi-compartmental Particles via the Micromolding Method (미세 성형 방법을 이용한 형광 표지된 이중 분획 입자의 제조)

  • Shim, Gyurak;Jeong, Seong-Geun;Hong, Woogyeong;Kang, Koung-Ku;Lee, Chang-Soo
    • Korean Chemical Engineering Research
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    • v.56 no.6
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    • pp.826-831
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    • 2018
  • This study presents fabrication of bi-compartmental particles labeled by multiple fluorescence. To compartmentalize fluorescent expression at the particle, two fluorescent dyes with less overlap of the excitation and emission spectra are selected. To ensure the fluorescence stability, the fluorescent dyes contain acrylate functional groups in the molecules so that they can be cross-linked together with monomers constituting the particle. Strong fluorescent expression and compartmentalization were observed at the particle fabricated using the selected fluorescent dyes through confocal microscopy. Furthermore, long-term fluorescence stability was verified by measuring fluorescent expression and intensity for 4 weeks. We anticipate that the bi-compartmental particles labeled by multiple fluorescence can be widely used for multi-target drug delivery system, analysis of 3 dimensional Brownian motion, and investigation of 3 dimensional complex self-assembled morphologies.

Anti-atopic dermatitis effects of Parasenecio auriculatus via simultaneous inhibition of multiple inflammatory pathways

  • Kwon, Yujin;Cho, Su-Yeon;Kwon, Jaeyoung;Hwang, Min;Hwang, Hoseong;Kang, Yoon Jin;Lee, Hyeon-Seong;Kim, Jiyoon;Kim, Won Kyu
    • BMB Reports
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    • v.55 no.6
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    • pp.275-280
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    • 2022
  • The treatment of atopic dermatitis (AD) is challenging due to its complex etiology. From epidermal disruption to chronic inflammation, various cells and inflammatory pathways contribute to the progression of AD. As with immunosuppressants, general inhibition of inflammatory pathways can be effective, but this approach is not suitable for long-term treatment due to its side effects. This study aimed to identify a plant extract (PE) with anti-inflammatory effects on multiple cell types involved in AD development and provide relevant mechanistic evidence. Degranulation was measured in RBL-2H3 cells to screen 30 PEs native to South Korea. To investigate the anti-inflammatory effects of Parasenecio auriculatus var. matsumurana Nakai extract (PAE) in AD, production of cytokines and nitric oxide, activation status of FcεRI and TLR4 signaling, cell-cell junction, and cell viability were evaluated using qRT-PCR, western blotting, confocal microscopy, Griess system, and an MTT assay in RBL-2H3, HEK293, RAW264.7, and HaCaT cells. For in vivo experiments, a DNCBinduced AD mouse model was constructed, and hematoxylin and eosin, periodic acid-Schiff, toluidine blue, and F4/80-staining were performed. The chemical constituents of PAE were analyzed by HPLC-MS. By measuring the anti-degranulation effects of 30 PEs in RBL-2H3 cells, we found that Paeonia lactiflora Pall., PA, and Rehmannia glutinosa (Gaertn.) Libosch. ex Steud. show an inhibitory activity of more than 50%. Of these, PAE most dramatically and consistently suppressed cytokine expression, including IL-4, IL-9, IL-13, and TNF-α. PAE potently inhibited FcεRI signaling, which mechanistically supports its basophil-stabilizing effects, and PAE downregulated cytokines and NO production in macrophages via perturbation of toll-like receptor signaling. Moreover, PAE suppressed cytokine production in keratinocytes and upregulated the expression of tight junction molecules ZO-1 and occludin. In a DNCB-induced AD mouse model, the topical application of PAE significantly improved atopic index scores, immune cell infiltration, cytokine expression, abnormal activation of signaling molecules in FcεRI and TLR signaling, and damaged skin structure compared with dexamethasone. The anti-inflammatory effect of PAE was mainly due to integerrimine. Our findings suggest that PAE could potently inhibit multi-inflammatory cells involved in AD development, synergistically block the propagation of inflammatory responses, and thus alleviate AD symptoms.