• BMB Reports
• /
• v.38 no.3
• /
• pp.354-359
• /
• 2005

Open reading frame 29 (ha29) is a gene specific for Helicoverpa armigera single-nucleocapsid nucleopolyhedrovirus (HearSNPV). Sequence analyses showed that the transcription factor Tfb2 motif, bromodomain and Half-A-TPR (HAT) repeat were present at aa 66-82, 4-76, 55-90 of the Ha29 protein respectively. The product of Ha29 was detected in HearSNPV-infected HzAM1 cells at 3 h post-infection. Western blot analysis using a polyclonal antibody produced by immunizing a rabbit with purified GST-Ha29 fusion protein indicates that Ha29 is an early gene. The size of Ha29 product in infected HzAM1 cells was about 25 kDa, which was larger than the presumed size of 20.4 kDa. Tunicamycin treatment of HearSNPV-infected HzAM1 cells suggested that the Ha29 protein is N-glycosylated. Fluorescent confocal laser scanning microscope examination, and Western blot analysis of purified budded virus (BVs), occlusion-derived virus (ODVs), cell nuclear and cytoplasmic fraction, showed that the Ha29 protein was localized in the nucleus. Our results suggested that ha29 of HearSNPV encodes a non-structurally functional protein that may be associated with virus gene transcription in Helicoverpa hosts.

• Korean Journal of Microbiology
• /
• v.52 no.2
• /
• pp.125-139
• /
• 2016

Biofilms are considered a complexly structured community of microorganisms derived from their attached growth to abiotic and biotic surfaces. In human life, they mediate serious infections and cause many problems in civil and industrial facilities. While it is of huge interest for scientists to understand biofilms, it has been very hard to directly analyze the various biofilms in nature. A variety of biofilm models have been suggested for laboratory-scale biofilm formation and many methods based on these models are widely used for the biofilm researches. These biofilm models mimic characteristics of environmental biofilms with different advantages and disadvantages. In this review, we will introduce these currently used biofilm model systems and explain their relative merits.

• Restorative Dentistry and Endodontics
• /
• v.43 no.1
• /
• pp.2.1-2.10
• /
• 2018

Objectives: To determine the effect of size and insertion depth of irrigation needle on the amount of apical extruded debris and the amount of penetration depth of sealer using a confocal laser scanning microscope (CLSM). Materials and Methods: Twenty maxillary premolars were assigned to 2 groups (n = 10), according to the size of needle tip, 28 G or 30 G. Buccal roots of samples were irrigated with respective needle type inserted 1 mm short of the working length (WL), while palatal roots were irrigated with respective needle type inserted 3 mm short of the WL. Prepared teeth were removed from the pre-weighed Eppendorf tubes. Canals were filled with F3 gutta-percha cone and rhodamine B dye-labeled AH 26 sealer. Teeth were transversally sectioned at 1 and 3 mm levels from the apex and observed under a CLSM. Eppendorf tubes were incubated to evaporate the irrigant and were weighed again. The difference between pre- and post-weights was calculated, and statistical evaluation was performed. Results: Inserting needles closer to the apex and using needles with wider diameters were associated with significantly more debris extrusion (p < 0.05). The position of needles and level of sections had statistically significant effects on sealer penetration depth (p < 0.05 for both). Conclusions: Following preparation, inserting narrower needles compatible with the final apical diameter of the prepared root canal at 3 mm short of WL during final irrigation might prevent debris extrusion and improve sealer penetration in the apical third.

• Journal of Life Science
• /
• v.22 no.10
• /
• pp.1415-1419
• /
• 2012

Thymosin ${\beta}4$ ($T{\beta}4$) has been reported to be overexpressed in CD133-positive colorectal cancer stem cells. We analyzed the relationship between $T{\beta}4$ and CD133-positive stem cells in normal stomach by examining the expression patterns of $T{\beta}4$ and CD133 in normal stomach tissues by immunohistochemical staining; co-localization of $T{\beta}4$ and CD133 was studied by immunofluorescence and confocal laser-scanning microscopy. Both $T{\beta}4$ and CD133 were expressed in stomach glands and showed similar expression patterns. Immunofluorescence staining of $T{\beta}4$ and CD133 showed that the expression of $T{\beta}4$ and CD133 was co-localized. In summary, both $T{\beta}4$ and CD133 were expressed in glands of normal stomachs and expression patterns were co-localized. These data suggest that $T{\beta}4$ expression is strongly related to CD133 expression.

• Journal of the Korea Convergence Society
• /
• v.8 no.6
• /
• pp.139-145
• /
• 2017

The aim of this study was to measure the remineralization effect of APF gel and fluride varnish on artificial enamel caries using CLSM in vitro. The samples were divided into 3 groups: control, 1.23% APF gel, 5% NaF varnish. The specimen surfaces were observed by CLSM and measured average fluorescence of the lesion(AFL). The results were analyzed using one-way ANOVA and Pearson's correlation analysis at a significance level off 0.05(PSWA 18.0, SPSS Inc., USA). There were significant differences between AFL at baseline and 1 day after fluoride application(p<0.05) but there are no significant differences between ${\Delta}$ AFL of all groups (p=0.222). Result of Pearson's correlation analysis, there are no significant correlation between VHN and AFL, but there were significant correlation between AFL at baseline and 1 day after fluoride application(r=0.811, p<0.001). Although AFL decreased after fluoride application, but there was no difference between the groups. In the future, it is necessary to test the oral environment model or in situ experiment supplemented the limitations of this study.

• The Journal of Korean Academy of Prosthodontics
• /
• v.44 no.3
• /
• pp.343-355
• /
• 2006

Statement of problem. The success of osseointegration can be enhanced with an implant that has improved surface characteristics. Anodic oxidation is one of the surface modifying method to achieve osseointegration. Voltage of anodic oxidation can change surface characteristics and cell activity Purpose. This study was performed to evaluate MG63 cell responses such as affinity, proliferation and to compare surface characteristics of anodic oxidized titanium in various voltage. Material and method. The disks for cell culture were fabricated from grade 3 commercially pure titanium,1 m in thickness and 12 mm in diameter. Surfaces of 4 different roughness were prepared. Group 1 had a machined surface, used as control. Group 2 was anodized under 220 V, group 3 was anodized under 300 V and group 4 was anodized under 320 V. The microtopography of specimens was observed by scanning electron microscope (JSM-840A, JEOL, Japan) and atomic force microscope(Autoprobe CP, Park Scientific Instrument, USA). The surface roughness was measured by confocal laser scanning microscope(Pascal, LSM5, Zeiss, Germany). The crystal structure of the titanium surface was analyzed with x-ray diffractometer(D8 advanced, Broker, Germany). MG63 osteoblast-like cells were cultured on these specimens. The cell morpholgy was observed by field emission electron microscope(Hitachi S-4700, Japan). The cell metabolic and proliferative activity was evaluated by MTT assay Results and conclusion. With in limitations of this in vitro study, the following conclusions were drawn. 1. In anodizing titanium surface, we could see pores which did not show in control group. In higher anodizing voltage, pore size was increased. 2. In anodizing titanium surface, we could see anatase. In higher anodizing voltage, thicker oxide layer increased crystallinity(anatase, anatase and rutile mixed). 3. MG63 cells showed more irregular, polarized and polygonal shape and developed more lamellipodi in anodizing group as voltage increased. 4. The activity of cells in MTT assay increased significantly in group 3 and 4 in comparison with group 1 and 2. However, there was no difference between group 3 and 4 at P<0.05. Proliferation of MG63 cells increased significantly in pore size($3-5.5{\mu}m$) of group 3 and 4 in comparison with in pore size($0.2-1{\mu}m$ ) of group 2.

• Journal of Pharmacopuncture
• /
• v.11 no.2
• /
• pp.5-12
• /
• 2008

Objective In this article, we report on the intradermal Alcian blue staining method for tracing the meridians of acupuncture. Methods 1% Alcian blue solution was injected into acupoints by using a 0.5mL insulin syringe with a 31-gauge needle, then the skin was incised and was observed under a stereoscopic microscope. The specimens were examined by using immunohistochemical methods and were observed under a confocal laser scanning microscope. Results A threadlike structure, which was visualized with Alcian blue, existed in dermis layer and proceeded to hypodermis. In this structure, characteristic alignments of rod- shaped nuclei and $1-2{\mu}m$ sized DNA granules were observed. Furthermore, abundant blood capillary plexuses, peripheral nerve endings, and a corpuscle-like structure(about $300{\mu}m$ in diameter) were visualized in the skin tissues of acupoints. Conclusion It was concluded that the specific threadlike and corpuscle-like structures corresponded to superficial Bonghan duct and corpuscle, respectively.

• The Journal of Advanced Prosthodontics
• /
• v.10 no.2
• /
• pp.147-154
• /
• 2018

PURPOSE. This study was performed to evaluate the osteogenic potential of 3mol% yttria-stabilized tetragonal zirconia polycrystals (3Y-TZP) and niobium oxide containing Y-TZPs with specific ratios, new (Y,Nb)-TZPs, namely YN4533 and YN4533/Al20 discs. MATERIALS AND METHODS. 3Y-TZP, YN4533 and YN4533/Al20 discs (15 mm diameter and 1 mm thickness) were prepared and their average surface roughness ($R_a$) and surface topography were analyzed using 3-D confocal laser microscope (CLSM) and scanning electron microscope (SEM). Mouse pre-osteoblast MC3T3-E1 cells were seeded onto all zirconia discs and evaluated with regard to cell attachment and morphology by (CLSM), cell proliferation by PicoGreen assay, and cell differentiation by Reverse-Transcription PCR and Quantitative Real-Time PCR, and alkaline phosphatase (Alp) staining. RESULTS. The cellular morphology of MC3T3-E1 pre-osteoblasts was more stretched on a smooth surface than on a rough surface, regardless of the material. Cellular proliferation was higher on smooth surfaces, but there were no significant differences between 3Y-TZP, YN4533, and YN4533/Al20. Osteoblast differentiation patterns on YN4533 and YN4533/Al20 were similar to or slightly higher than seen in 3Y-TZP. Although there were no significant differences in bone marker gene expression (alkaline phosphatase and osteocalcin), Alp staining indicated better osteoblast differentiation on YN4533 and YN4533/Al20 compared to 3Y-TZP. CONCLUSION. Based on these results, niobium oxide containing Y-TZPs have comparable osteogenic potential to 3Y-TZP and are expected to be suitable alternative ceramics dental implant materials to titanium for aesthetically important areas.

• Journal of dental hygiene science
• /
• v.23 no.4
• /
• pp.369-377
• /
• 2023

Background: In this study, we investigated the potential use of keratin for oral tissue regeneration. Keratin is well-known for its effectiveness in skin regeneration by promoting keratinization and enhancing the elasticity and activity of fibroblasts. Because of its structural stability, high storability, biocompatibility, and safety in humans, existing research has predominantly focused on its role in skin wound healing. Herein, we propose using keratin proteins as biocompatible materials for dental applications. Methods: To assess the suitability of alpha-keratin protein as a substrate for cell culture, keratin was extracted from human hair via PEGylation. Viabilities of primary human gingival fibroblasts (HGFs) and human oral keratinocytes (HOKs) were assessed. Fluorescence immunostaining and migration assays were conducted using a fluorescence microscope and confocal laser scanning microscope. Wound healing and migration assays were performed using automated software to analyze the experimental readout and gap closure rate. Results: We confirmed the extraction of alpha-keratin and formation of the PEG-g-keratin complex. Treatment of HGFs with keratin protein at a concentration of 5 mg/ml promoted proliferation and maintained cell viability in the test group compared to the control group. HOKs treated with 5 mg/ml keratin exhibited a slight decrease in cell proliferation and activity after 48 hours compared to the untreated group, followed by an increase after 72 hours. Wound healing and migration assays revealed rapid closure of the area covered by HOKs over time following keratin treatment. Additionally, HOKs exhibited changes in cell morphology and increased the expression of the mesenchymal marker vimentin. Conclusion: Our study demonstrated the potential of hair keratin for soft tissue regeneration, with potential future applications in clinical settings for wound healing.

• The Journal of Engineering Geology
• /
• v.14 no.1
• /
• pp.47-60
• /
• 2004

The permeability coefficients were calculated by the homogenization analysis method with sufficient consideration of fracture geometry dependent on aperture change. According to the results of aperture measurements using a confocal laser scanning microscope, apertures on each measuring point display different magnitudes, indicating that fracture walls can not be assumed as parallel feature. After construction of fracture model based on the aperture values measured on each pressure level, the homogenization analysis was conducted to compute permeability coefficients. The calculated permeability coefficients distribute in the ranges of $10^{-1}~10^{-3}cm/sec$. Most of the specimens show decreasing permeability coefficients with the increase of the applied pressure. However, the decreasing rates of permeability coefficients do not show a constant trend on each pressure level. This phenomenon is well matched to the observation results of Chae et al. (2003). It proves that aperture change strongly influences on permeability characteristics. Three sections of each specimen have all different values of permeability coefficient. It suggests that the variation of permeability coefficient depends sensitively on aperture magnitudes and characteristics of fracture geometry. It is very important to consider accurate fracture geometries for analysis of permeability characteristics in rock fractures bearing different aperture distribution. Therefore, it needs to consider sufficiently the fracture geometries for calculating the permeability coefficients of fractures.