• Journal of the korean academy of Pediatric Dentistry
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• v.28 no.1
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• pp.25-31
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• 2001

The purpose of this study was to investigate the distribution and fluorescence intensity of vasoactive intestinal polypeptide immunoreactive (VIP-IR) cells in rat trigeminal ganglion following pulp extirpation of rat mandibular molar. The animals were divided into control group(n=6) and experimental group(n=6). The experimental animals were sacrificed at 14 days after pulp extirpation. The trigeminal ganglion was removed and immersed in the 4% paraformaldehyde in 0.1M phosphate buffer. Serial frozen sections about $20{\mu}m$ in thickness were cut with a cryostat. The immunofluorescence staining was performed. The rabbit anti-VIP(1 : 8,000) was used as primary antibody and fluorescene isothiocynate(FITC) conjugated anti-rabbit IgG(1 : 80) as secondary antibody. The slides were observed under confocal laser scanning microscope (CLSM). Unprocessed optical sections were obtained and stored on a optical disk. Color pictures were printed by a video copy processor. The results were as follows; 1. The positive ratio of VIP-IR cells in mandibular part of trigeminal ganglion were 7.40% in control group and 28.42% in experimental group(14 days after pulp extirpation). 2. The relative fluorescence intensity of VIP-IR cells in mandibular part of trigeminal ganglion were 87.78 in control group and 138.65 in experimental group. The relative fluorescence intensity of experimental group was 58% higher than that of control group. 3. In optical serial section analysis of VIP-IR cells of experimental group, most of the 9 sections showed high fluorescence intensity. At high magnification, axons of the experimental group displayed greater VIP-IR than in the control group, and the positive cells were mainly of medium size. The result indicate that number and fluorescence intensity of VIP-IR cells were increased in the mandibular part of trigeminal ganglion following pulp extirpation of mandibular molar, and it suggests that VIP could play a role in processing of nociception.

• Childhood Kidney Diseases
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• v.15 no.2
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• pp.138-145
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• 2011

Purpose : To test whether the expression of ${\beta}$-catenin, a component of podocyte as a filtration molecule, would be altered by puromycin aminonucleoside (PAN) in the cultured podocyte in vitro. Methods : We cultured rat glomerular epithelial cells (GEpC) with various concentrations of PAN and examined the distribution of ${\beta}$-catenin by confocal microscope and measured the change of ${\beta}$-catenin expression by Western blotting and reverse transcriptase-polymerase chain reaction (RT-PCR). Results :We found that ${\beta}$-catenin relocalized from peripheral cytoplasm to inner cytoplasm, therefore, intercellular separations were seen in confluently cultured cells by high concentrations of PAN in immunofluorescence views. In Western blotting of GEpC, PAN ($50{\mu}g/mL$) decreased ${\beta}$-catenin expression by 34.9% at 24 hrs and 34.3% at 48 hrs, compared to those in without PAN condition (P<0.05). In RT-PCR, high concentrations ($50{\mu}g/mL$) of PAN also decreased ${\beta}$-catenin mRNA expression similar to protein suppression by 25.4% at 24 hrs and 51.8% at 48 hrs (P<0.05). Conclusion : Exposure of podocytes to PAN in vitro relocates ${\beta}$-catenin internally and reduces ${\beta}$-catenin mRNA and protein expression, which could explain the development of proteinuria in experimental PAN-induced nephropathy.

• Applied Microscopy
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• v.34 no.4
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• pp.231-239
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• 2004

Cerebral microvessel endothelial cells that form blood-brain barrier (BBB) have tight junction for maintaining brain homeostasis. Occludin, one of tight junction protein, is crucial for BBB function. $H_2O_2$ induced occludin changes and effects in bovine brain BBB endothelial cells were examined in this study. The decrease of transendothelial electrical resistance (TEER) by $H_2O_2$ was due to disruption of occludin localization. Cytotoxicity test revealed that $H_2O_2$ did not cause cell death below 1 mM $H_2O_2$ within 4 hr. $H_2O_2$ caused intermittent disruption and loss of occludin at tight junctions and occludin disappeared with dose dependent manner from tight junction in confocal laser microscopy. But Western blot revealed that the total amounts of occludin increased by $H_2O_2$ administration. Transmission electron microscopy revealed that the ultrastructure of tight junction was not changed by $H_2O_2$. These data suggest that functional disruption of BBB by $H_2O_2$ was due to the localized loss of occludin in tight junction, but the expression of occludin increased in order to compensate the disrupted function in BBB.

• Journal of Korean Academy of Oral and Maxillofacial Radiology
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• v.29 no.2
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• pp.435-449
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• 1999

Purpose : The study was aimed to detect the differences in the cell viability and the apoptosis induction after irradiation on normal and tumorigenic cells. Materials and Methods : The study. that was generated for two human normal cells(RHEK, HGF-l) and two human tumor cells(KB. HT-1080). was tested using MTT assay at 1 day and 3 day after irradiation and TUNEL assay under confocal laser scanning microscope at 1 day after irradiation. Single irradiation of 0.5. 1, 2. 4. and 8Gy were applied to the cells. The two fractions of 1. 2. 4. and 8Gy were separated with a 4-hour time interval. The irradiation was done with 5.38Gy/min dose rate using Cs-137 irradiator at room temperature. Results and Conclusions : 1. In 3-day group. the cell viability of HGF-1 cell was significantly decreased at 2. 4 and 8Gy irradiation, the cell viability of KB cell was significantly decreased at 8Gy irradiation and the cell viability of HT-I080 cell was significantly decreased at 4 and 8Gy irradiation. 2. There was significant difference between RHEK and KB cell line in the cell viability of 3-day group at 8Gy irradiation. There was significant difference between RHEK and HGF-1 cell line in the cell viability of 3-day group at 4 and 8Gy irradiation. 3. There was a significantly decreased cell viability in 3-day group than those in 1-day group at 2. 4 and 8Gy on HGF-1 cell. at 4 and 8Gy on HT-I080 cell. at 8Gy on KB cell. 4. We could detect DNA fragmented cells only on KB cell. Number of apoptotic cells of KB cell was significantly increased at 4 and 8Gy irradiation. However, there was no correlation between cell viability and apoptosis. 5. On all 4 cell lines, there were no differences between single and split irradiation method in cell viability and apoptosis.

• Journal of the korean academy of Pediatric Dentistry
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• v.45 no.3
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• pp.290-298
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• 2018

This study aimed to evaluate surface morphology and resin tag penetration of resin infiltration into primary anterior teeth after enamel deproteinization with sodium hypochlorite (NaOCl) prior to phosphoric acid ($H_3PO_4$) etching. Ninety primary anterior teeth with non-cavitated caries lesion were devided five groups according to enamel pretreatment as follows, group I-15% hydrochloric acid (HCl) 2min. ; group II-5.25% NaOCl 1min., 35% $H_3PO_4$ 1min. ; group III-5.25% NaOCl 2min., 35% $H_3PO_4$ 1min. ; group IV-5.25% NaOCl 1min., 35% $H_3PO_4$ 2min. ; group V-5.25% NaOCl 2min., 35% $H_3PO_4$ 2min. Fifteen teeth were examined etched surface structure using field emission-scanning electron microscope. Seventy five teeth were infiltrated with resin, maximum penetration depth and percentage penetration were analysed using dual fluorescence confocal microscopy. As the application time of NaOCl increased, ratio of enamel type I, II were increased. Percentage penetration (PP) was higher in group V than group II, III (p < 0.05). PP of group IV, V did not show any differences. Non-cavitated caries of primary anterior teeth can be treated with resin infiltration. Enamel deproteinization with NaOCl prior to 35% $H_3PO_4$ etching could be an alternative of 15% HCl etching in resin infiltration.

• The korean journal of orthodontics
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• v.36 no.5
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• pp.360-368
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• 2006

Objective: Surface characteristics of dental materials play an important role in bacterial adhesion. The purpose of this study was to investigate surface characteristics of 5 different light-cured orthodontic adhesives (1 fluoride-releasing composite, 3 non-fluoride-releasing composites, and f resin-modified glass ionomer). Methods: Surface roughness was measured using a confocal laser scanning microscope. Contact angle and surface free energy components were analyzed using the sessile drop method. Results: Surface roughness was significantly different between adhesives despite a relatively small variation (less than $0.05\;{\mu}m$). Lightbond and Monolok2 were rougher than Enlight and Transbond XT. There were also significant differences in contact angles and surface free energy components between adhesives. In particular, considerable differences in contact angles and surface free energy components were found between resin modified glass ionomer and the composites. Resin modified glass ionomer showed significantly smaller contact angles in 3 different probe liquids and had higher total surface free energy and stronger polarity, with notably stronger basic property than the composites. Conclusion: Resin modified glass ionomer may provide a more favourable environment for bacterial adhesion than composite adhesives.

• Journal of Conservation Science
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• v.30 no.2
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• pp.123-136
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• 2014

Considering that decision in conservation treatment for damaged objects should consider not only various options of methodology of intervention but also possible consequences of different types of intervention, it is a difficult task to decide and propose clear and safest solution of preserving an object. In addition, it should be constantly challenged by conservators even if it is proved technique or methodology in a past treatment. Therefore, there is no absolute solution which can be applied to all practice but each decision can be different case by case. It is not possible to estimate the way how the present condition of material and environmental aspects would affect to the condition of an object in future. However if conservators keep trying to set out various ways of analysing pro and against effect of past treatments, it would be able to provide useful logics of proving efficiency and appropriateness of a certain treatment. Understanding that the advantage of laser technique is to adopt a way of cleaning an object without making a direct contact, which is different from other techniques, this paper aims at securing stability of laser techniques, although it remains a limitation in the compatibility to all other materials. This study has examined reacting process on the painted pigments against laser beam by using CLSM in order for it to display both the problems from such reacting process and the efficiency of it as a cleaning methodology. It has intended to estimate the result of laser techniques and propose the range of applicability.

• Korean Journal of Optics and Photonics
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• v.19 no.2
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• pp.87-94
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• 2008

We present a new design for an endoscope objective lens composed of a lexible fiber bundle with 30,000 core, and a gradient-index (GRIN) objective lens with an optical adaptor. The characteristic of this objective lens is to be minimally-invasive to be able to insert easily in the internal organs of live animals. The GRIN lens has a small diameter and a very simple construction, which is selected with the diameter of 1.0 mm and numerical aperture of 0.5 to achieve a minimally-invasive outer diameter and a high resolution. The resultant designed lens shows the performance as follows; a lateral resolution of 1.63 um and diameters of 100% encircled energy of $0.3\;{\mu}m$ and $0.83\;{\mu}m$ for the on-axis and the off-axis image point, respectively. Also, we can present a cheap solution with a lateral resolution of 1.74 um and diameters of 100% encircled energy of $1.10\;{\mu}m$ and $2.84\;{\mu}m$ for the on-axis and the off-axis image point, respectively.

• Journal of Periodontal and Implant Science
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• v.44 no.2
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• pp.79-84
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• 2014

Purpose: While single-species biofilms have been studied extensively, we know notably little regarding multispecies biofilms and their interactions. The purpose of this study was to develop and evaluate an in vitro multispecies dental biofilm model that aimed to mimic the environment of chronic periodontitis. Methods: Streptococcus gordonii KN1, Fusobacterium nucleatum ATCC23726, Aggregatibacter actinomycetemcomitans ATCC33384, and Porphyromonas gingivalis ATCC33277 were used for this experiment. The biofilms were grown on 12-well plates with a round glass slip (12 mm in diameter) with a supply of fresh medium. Four different single-species biofilms and multispecies biofilms with the four bacterial strains listed above were prepared. The biofilms were examined with a confocal laser scanning microscope (CLSM) and scanning electron microscopy (SEM). The minimum inhibitory concentrations (MIC) for four different planktonic single-species and multispecies bacteria were determined. The MICs of doxycycline and chlorhexidine for four different single-species biofilms and a multispecies biofilm were also determined. Results: The CLSM and SEM examination revealed that the growth pattern of the multispecies biofilm was similar to those of single-species biofilms. However, the multispecies biofilm became thicker than the single-species biofilms, and networks between bacteria were formed. The MICs of doxycycline and chlorhexidine were higher in the biofilm state than in the planktonic bacteria. The MIC of doxycycline for the multispecies biofilm was higher than were those for the single-species biofilms of P. gingivalis, F. nucleatum, or A. actinomycetemcomitans. The MIC of chlorhexidine for the multispecies biofilm was higher than were those for the single-species biofilms of P. gingivalis or F. nucleatum. Conclusions: To mimic the natural dental biofilm, a multispecies biofilm composed of four bacterial species was grown. The 24-hour multispecies biofilm may be useful as a laboratory dental biofilm model system.

• Journal of Periodontal and Implant Science
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• v.47 no.4
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• pp.219-230
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• 2017

Purpose: The purpose of this study was to compare the characteristics of single- and dualspecies in vitro oral biofilms made by static and dynamic methods. Methods: Hydroxyapatite (HA) disks, 12.7 mm in diameter and 3 mm thick, were coated with processed saliva for 4 hours. The disks were divided into a static method group and a dynamic method group. The disks treated with a static method were cultured in 12-well plates, and the disks in the dynamic method group were cultured in a Center for Disease Control and Prevention (CDC) biofilm reactor for 72 hours. In the single- and dual-species biofilms, Fusobacterium nucleatum and Porphyromonas gingivalis were used, and the amount of adhering bacteria, proportions of species, and bacterial reduction of chlorhexidine were examined. Bacterial adhesion was examined with scanning electron microscopy (SEM) and confocal laser scanning microscopy (CLSM). Results: Compared with the biofilms made using the static method, the biofilms made using the dynamic method had significantly lower amounts of adhering and looser bacterial accumulation in SEM and CLSM images. The proportion of P. gingivalis was higher in the dynamic method group than in the static method group; however, the difference was not statistically significant. Furthermore, the biofilm thickness and bacterial reduction by chlorhexidine showed no significant differences between the 2 methods. Conclusions: When used to reproduce periodontal biofilms composed of F. nucleatum and P. gingivalis, the dynamic method (CDC biofilm reactor) formed looser biofilms containing fewer bacteria than the well plate. However, this difference did not influence the thickness of the biofilms or the activity of chlorhexidine. Therefore, both methods are useful for mimicking periodontitis-associated oral biofilms.