• Restorative Dentistry and Endodontics
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• 제28권5호
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• pp.392-401
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• 2003

The purpose of this study was to evaluate the penetration pattern of dentin adhesives according to the orientation of dentinal tubules with confocal laser scanning microscopy. Specimens having perpendicular. parallel and oblique surface to dentinal tubules were fabricated. The primer of dentin adhesives (ALL $BOND^{\circledR}{\;}2,{\;}CLEARFIL^{TM}$ SE BOND and PQ1) was mixed with fluorescent material. rhodamine B isothio-cyanate (Aldrich Cherm. CO., Milw., USA), It was applied to the specimens according to the instructions of manufactures. The specimens were covered with composite resin (Estelite, shade A2) and then cut to a thickness of 500$\mu\textrm{m}$ with low speed saw (Isomet^{TM}, Buehler. USA). The adhesive pattern of dentin adhesives were observed by fluorescence image using confocal laser scanning microscopy. The results were as follows. 1. For the groups with tubules perpendicular to bonded surface. funnel shape of resin tag was observed in all specimen. However. resin tags were more prominent in phosphoric acid etching system (ALL $BOND^{\circledR}$ 2 and PQ1) than self etching system ($CLEARFIL^{TM}$ SE BOND). 2. For the groups with tubules parallel to bonded surface. rhodamine-labeled primer penetrated into peritubular dentin parallel to the orientation of dentinal tubules. But rhodamine-labeled primer of PQ1 diffused more radially into surrounding intertubular dentin than other dentin adhesive systems. 3. For the groups with tubules oblique to bonded surface. resin tags appeared irregular and discontinuous. But they penetrated deeper into dentinal tubules than other groups.

• 한국산학기술학회논문지
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• 제20권5호
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• pp.1-6
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• 2019

유기 염료가 도핑 된 실리카 나노입자는 바이오 라벨링, 바이오 이미징 및 바이오 센싱에 사용되고 있는 유망한 나노소재이다. 일반적으로 형광 실리카 나노입자는 수정된 스토버 방법($St{\ddot{o}}ber$ Method)으로 합성된다. 본 연구에서는 다양한 크기를 갖는 염료가 첨가되지 않은 형광 실리카 나노입자를 수정된 스토버 합성법인 졸겔 공정으로 합성하였다. 졸겔 공정 중에 기능성 물질인 APTES를 첨가제로 첨가하였다. 졸겔 공정으로 합성된 실리카 나노입자는 $400^{\circ}C$에서 2시간 동안 하소되었다. 합성된 실리카 나노입자의 표면형상과 크기를 전계방출 주사전자현미경으로 조사하였고, 합성된 실리카 나노입자의 형광 특성은 파장 365 nm의 자외선 램프를 조사하여 확인하였다. 또한 합성된 실리카 나노입자의 광발광 (PL) 특성을 형광 분석 형광법으로 조사하였다. 그 결과 합성된 실리카 나노입자는 입자의 크기와 무관하게 모두 청색 형광 특성을 갖는 것으로 확인되었다. 특히, 실리카 나노입자의 크기가 증가할수록 PL 강도는 감소하였다. 염료가 첨가되지 않은 실리카 나노입자의 청색 형광 특성은 APTES 층의 $NH_2$ 기능기와 실리카 매트릭스 뼈대 내부의 산소관련 결함과의 결합에 기인하는 것으로 추정된다.

• Journal of Microbiology and Biotechnology
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• 제19권5호
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• pp.495-501
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• 2009

A new spore display method is presented that enables recombinant proteins to be displayed on the surface of Bacillus spores via fusion with InhA, an exosporium component of Bacillus thuringiensis. The green fluorescent protein and $\beta$-galactosidase as model proteins were fused to the C-terminal region of InhA, respectively. The surface expression of the proteins on the spores was confirmed by flow cytometry, confocal laser scanning microscopy, measurement of the enzyme activity, and an immunogold electron microscopy analysis. InhA-mediated anchoring of foreign proteins in the exosporium of Bacillus spores can provide a new method of microbial display, thereby broadening the potential for novel applications of microbial display.

• Asian Pacific Journal of Cancer Prevention
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• 제13권6호
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• pp.2753-2758
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• 2012

Rubia cordifolia Linn, which belongs to the Rubiaceae family, is a well-known herb used in Ayurvedic medicine. In the present study, we investigated the influence of a methanolic extract (RC) on the induction of apoptosis in HEp-2 (human laryngeal carcinoma) cell line, as evidenced by cytotoxicity, morphological changes and modification in the levels of pro-oxidants. Inhibition of cell proliferation and lactate dehydrogenase (LDH) release increased in a time and dose-dependent manner. Further, reduced glutathione (GSH), glutathione transferase (GST) and protein levels decreased and lipid peroxidation increased significantly on RC treatment in a dose dependent manner when compared to controls. Based on the results we determined the optimal dose as 30mg/ ml and the apoptotic effect of RC extract (30 mg/ml) on HEp-2 cells was confirmed by fluorescent microscopy and transmission electron microscopy (TEM) based on morphological and ultrastructural changes. RC extract suppressed the proliferation of HEp-2 oral cancer cells inducing apoptotic cell death in vitro. These results point to potential of RC extract as an agent for the treatment of laryngeal squamous cell carcinoma.

• International Journal of Oral Biology
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• 제40권2호
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• pp.103-109
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• 2015

Growing evidence suggests that mitochondrial reactive oxygen species (ROS) are involved in various pain states. This study was performed to investigate whether ROS-induced changes in neuronal excitability in trigeminal subnucleus caudalis are related to ROS generation in mitochondria. Confocal scanning laser microscopy was used to measure ROS-induced fluorescence intensity in live rat trigeminal caudalis slices. The ROS level increased during the perfusion of malate, a mitochondrial substrate, after loading of 2',7'-dichlorofluorescin diacetate ($H_2DCF-DA$), an indicator of the intracellular ROS; the ROS level recovered to the control condition after washout. When pre-treated with phenyl N-tert-butylnitrone (PBN) and 4-hydroxy-2,2,6,6-tetramethylpiperidene-1-oxyl (TEMPOL), malate-induced increase of ROS level was suppressed. To identify the direct relation between elevated ROS levels and mitochondria, we applied the malate after double-loading of $H_2DCF-DA$ and chloromethyl-X-rosamine (CMXRos; MitoTracker Red), which is a mitochondria-specific fluorescent probe. As a result, increase of both intracellular ROS and mitochondrial ROS were observed simultaneously. This study demonstrated that elevated ROS in trigeminal subnucleus caudalis neuron can be induced through mitochondrial-ROS pathway, primarily by the leakage of ROS from the mitochondrial electron transport chain.

• 한국미생물·생명공학회지
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• 제47권2호
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• pp.303-309
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• 2019

Earlier studies by our group revealed that gallic acid in phytochemicals stimulated biofilm production in epiphytes, while caffeic acid in phytochemicals inhibited biofilm production in non-epiphytes. It is well documented that antimicrobial secretion by some epiphytic bacteria inhibits non-epiphytic bacterial growth on leaf surfaces. These selection criteria help plants choose their microbial inhabitants. Calcium and iron in phytochemicals also stimulate biofilm formation and thus, may be selection criteria adopted by plants with respect to their native epiphytic population. Furthermore, the processing of leaves during phytochemical extraction impacts the composition of the extract, and therefore its ability to affect bacterial biofilm formation. Computation of the Hurst exponent using biofilm thickness data obtained from the Ellipsometry of Brewster Angle Microscopic (BAM) images is an efficient tool for understanding the impact of phytochemicals on epiphytic and non-epiphytic populations when compared to fluorescent microscopy, scanning electron microscopy, and staining techniques. To the best of our knowledge, this is the first report that uses the Hurst exponent to elucidate the mechanism involved in plant microbe interaction.

• The Plant Pathology Journal
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• 제37권4호
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• pp.347-355
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• 2021

Ripe rot caused by Botryosphaeria dothidea is one of the serious diseases of postharvest kiwifruit. In order to control ripe rot on Actinidia chinensis cultivar 'Zesy002', several commercial agrofungicides were selected by an antifungal test on an artificial medium. Furthermore, disease suppression by the selected fungicides was evaluated on the kiwifruit by inoculation with a conidial suspension of B. dothidea. On the artificial media containing boscalid + fludioxonil was shown to be the most effective antifungal activity. However, in the bio-test pyraclostrobin + boscalid and iminoctadinetris were the most effective agrochemicals on the fruit. On the other hand, the infection structures of B. dothidea on kiwifruit treated with pyraclostrobin + boscalid were observed with a fluorescent microscope. Most of the fungal conidia had not germinated on the kiwifruit treated with the agrochemicals whereas on the untreated fruit the fungal conidia had mostly germinated. Electron microscopy of the fine structures showed morphological changes to the conidia and branch of hyphae on the kiwifruit pre-treated with pyraclostrobin + boscalid, indicating its suppression effect on fungal growth. Based on this observation, it is suggested that ripe rot by B. dothidea may be suppressed through the inhibition of conidial germination on the kiwifruit treated with the agrochemicals.

• Journal of the Optical Society of Korea
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• 제18권5호
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• pp.551-557
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• 2014

In this study, we demonstrated multimodal nonlinear optical (NLO) microscopy integrated simultaneously with two-photon excitation fluorescence (TPEF), second-harmonic generation (SHG), and coherent anti-Stokes Raman scattering (CARS) in order to obtain targeted cellular and label-free images in an immunofluorescence assay of the atherosclerotic aorta from apolipoprotein E-deficient mice. The multimodal NLO microscope used two laser systems: picosecond (ps) and femtosecond (fs) pulsed lasers. A pair of ps-pulsed lights served for CARS (817 nm and 1064 nm) and SHG (817 nm) images; light from the fs-pulsed laser with the center wavelength of 720 nm was incident into the sample to obtain autofluorescence and targeted molecular TPEF images for high efficiency of fluorescence intensity without cross-talk. For multicolor-targeted TPEF imaging, we stained smooth-muscle cells and macrophages with fluorescent dyes (Alexa Fluor 350 and Alexa Fluor 594) for an immunofluorescence assay. Each depth-sectioned image consisted of $512{\times}512$ pixels with a field of view of $250{\times}250{\mu}m^2$, a lateral resolution of $0.4{\mu}m$, and an axial resolution of $1.3{\mu}m$. We obtained composite multicolor images with conventional label-free NLO images and targeted TPEF images in atherosclerotic-plaque samples. Multicolor 3-D imaging of atherosclerotic-plaque structural and functional composition will be helpful for understanding the pathogenesis of cardiovascular disease.

• Journal of Plant Biotechnology
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• 제2권3호
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• pp.129-134
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• 2000

Clear visualization of pepper (Capsicum annuum L.) microspore nuclei with common stains such as acetocarmine or propionocarmine is difficult, hindering cytological analysis. The DAPI stain after the addition of ferric chloride solution to fixative resulted in clear visualization of nuclei. For clear visualization of nuclei and slight fluorescence of microspore wall, addition of 40-60 ${mu}ell$ of ferric chloride solution to the 1 $m\ell$ fixative was identified as most effective. At all stages of gametophytic development, the nuclei can be distinctly visualized. Starch granules does not intefere with the fluorochrome, and so the vegetative and generative nuclei were cleary visible in binucleate pollens. With its rapidity and reliability, this technique represents an efficient tool for routine staging or investigation of the nuclear status of the microspore during culture.

• 한국생물공학회:학술대회논문집
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• 한국생물공학회 2003년도 생물공학의 동향(XII)
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• pp.315-319
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• 2003

A novel method is developed to yield virus titers in 10 h, is easy to .perform using 96-well plates, and applicable to both any Autographa californica nucleopolyhyderovirus (AcNPV) and Bombyx mori nucleopolyhedrovirus (BmNPV)-based recombinant baculovirus. This assay uses an antibody to a DNA-binding protein to detect the infected cells via immune-staining. The titer is determined by counting foci produced due to infection of virus under a fluorescent microscopy. The required incubation period was shortened considerably because infected cells expressed viral antigens at the post infection time of 4 h. Therefore, 10 hours were enough to estimate the virus titer including virus infection time, insect cell culture, and estimation of virus titer.