• Title/Summary/Keyword: Confocal laser scanning microscope

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Development of Female Gametophyte of Six Species of Corydalis (Fumariaceae) (현호색속(현호색과) 6종의 자성배우자체 발달과정)

  • Oh, Byoung-Un;Jang, Chang-Gee
    • Korean Journal of Plant Taxonomy
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    • v.32 no.4
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    • pp.467-480
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    • 2002
  • The development of the female gametophyte of Corydalis albipetala, C. ambigua, C. filistipes, C. nobilis, C. solida, C. ophiocarpa have been comparatively investigated using laser scanning confocal microscope (LSCM) and light microscope. An archesporium was originated from one of the outmost parietal cells beneath the one-layered epidermis of protuberant nucellus, and acted directly as a megaspore mother cell (MMC). These species had linear tetrads after successional meiotic division during the megasprogenesis. A functional megasprore developed from one of the tetrad in the chalazal end, and the rest three being degenerated. The developmental type of the female gametophyte was monosporic in accordance with the Polygonum type. Prior to anthesis the female gametophyte was organized. So mature embryo sac was comprised a three-celled egg apparatus, three large antipodals were developed from the apex of each antipodal cell, and extended toward micropylar end to be contacted with egg apparatus. Two synergids were usually observed as degenerated condition, and in this time the apices of antipodal haustoria were connected with the degenerated synergids. The developmental characteristics of seven-nucleate female gametophytes were common in all the species investigated. But the shape of mature embryo sac was ovoidal in C. albipetala, C. filistipes, C. ophiocarpa and C. solida, reflexed in C. ambigua, and rather flattened ovoidal in C. nobilis. Also, the type of megasporangium was anatropous in all the species except C. ambigua with campylotropous ovule.

A periodontitis-associated multispecies model of an oral biofilm

  • Park, Jong Hwa;Lee, Jae-Kwan;Um, Heung-Sik;Chang, Beom-Seok;Lee, Si-Young
    • 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.

Comparison of periodontitis-associated oral biofilm formation under dynamic and static conditions

  • Song, Won sub;Lee, Jae-Kwan;Park, Se Hwan;Um, Heung-Sik;Lee, Si Young;Chang, Beom-Seok
    • 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.

Preparation and Characterization of Double-Layered Coated Capsule Containing Low Molecular Marine Collagen and γ-Aminobutyric Acid Producing Lactobacillus brevis CFM20 (저분자 해양성 콜라겐과 γ-Aminobutyric Acid 생성 Lactobacillus brevis CFM20을 함유하는 이중코팅캡슐의 제조 및 특성)

  • Kim, Sun-Yeong;Oh, Do-Geon;Kim, Kwang-Yup
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.46 no.7
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    • pp.857-867
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    • 2017
  • This study was performed to encapsulate low molecular weight marine collagen and ${\gamma}$-aminobutyric acid (GABA)-producing lactic acid bacteria to inhibit degradation and improve survival rate during exposure to adverse conditions of the gastro-intestinal tract. Calcium-alginate method was used for the manufacture of a double-layered coated capsule. The inner core material was composed of collagen and lactic acid bacteria, and the coating materials were alginate and chitosan. The sizes and shapes of the double-coated capsule were affected mainly by centrifuge speed and pH. Manufactured capsules were observed with a scanning electron microscope and by confocal laser scanning microscopy to confirm the micromorphological changes of capsules and bacterial cells. As a result, double-layered coated capsules were not degraded at pH 1.2, whereas degradation occurred at pH 7.4. In addition, GABA and collagen were maintained in stable state at pH 1.2. Therefore, double-layered coated capsules developed in this study would not be degraded in the stomach and could be stably delivered to the small intestine to benefit intestinal and dermatic health.

CHANGE OF VASOACTIVE INTESTINAL POLYPEPTIDE(VIP) IMMUNOREACTIVE CELLS FOLLOWING PULP EXTIRPATION IN RAT TRIGEMINAL GANGLION: A CONFOCAL LASER SCANNING MICROSCOPIC STUDY (치수제거 후 흰쥐 삼차신경절에서 VIP 면역반응세포의 변화: 공초점레이저주사현미경적 연구)

  • Kim, Heung-Joong;Kim, Seung-Jae;Park, Joo-Cheol;Lee, Chang-Seop;Lee, Sang-Ho
    • 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.

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Influence of finish line design on the marginal fit of nonprecious metal alloy coping fabricated by 3D printing, milling and casting using CAD-CAM (CAD-CAM을 이용한 3D printing, milling, casting 방법의 비귀금속 코핑의 지대치 변연 적합도 연구)

  • Seo-Rahng Kim;Myung-Joo Kim;Ji-Man Park;Seong-Kyun Kim;Seong-Joo Heo;Jai-Young Koak
    • The Journal of Korean Academy of Prosthodontics
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    • v.61 no.1
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    • pp.1-17
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    • 2023
  • Purpose. The purpose of this study was to examine the correlation between the finish line designs and the marginal adaptation of nonprecious metal alloy coping produced by different digital manufacturing methods. Materials and methods. Nonprecious metal alloy copings were made respectively from each master model with three different methods; SLS, milling and casting by computer aided design and computer aided manufacturing (CAD-CAM). Twelve copings were made by each method resulting in 72 copings in total. The measurement was conducted at 40 determined reference points along the circumferential margin with the confocal laser scanning microscope at magnification ×150. Results. Mean values of marginal gap of laser sintered copings were 11.8 ± 7.4 ㎛ for deep chamfer margin and 6.3 ± 3.5 ㎛ for rounded shoulder margin and the difference between them was statistically significant (P < .0001). Mean values of marginal gap of casted copings were 18.8 ± 20.2 ㎛ for deep chamfer margin and 33 ± 20.5 ㎛ for rounded shoulder margin and the difference between them was significant (P = .0004). Conclusion. Within the limitation of this study, the following conclusions were drawn. 1. The variation of finish line design influences the marginal adaptation of laser sintered metal coping and casted metal coping. 2. Laser sintered copings with rounded shoulder margin had better marginal fit than deep chamfer margin. 3. Casted copings with deep chamfer margin had better marginal fit than rounded shoulder margin. 4. According to the manufacturing method, SLS system showed the best marginal fit among three different methods. Casting and milling method followed that in order.

Effect of Radiation Dosage Changes on the Cell Viability and the Apoptosis Induction on Normal and Tumorigenic Cells (방사선의 선량변화가 수종의 정상세포와 종양세포주의 세포활성도와 apoptosis 유발에 미치는 영향)

  • Park In-Woo;Lee Sam-Sun;Heo Min-Suk;Choi Soon-Chul
    • 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.

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Surface roughness and surface free energy components of various orthodontic adhesives (다양한 교정용 접착제의 표면거칠기와 표면에너지 요소 분석)

  • Ahn, Hyo-Beom;Ahn, Sug-Joon;Nahm, Dong-Seok
    • 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.

The Effect of Silver Diamine Fluoride on Salivary Biofilm (Silver diamine fluoride가 타액 생물막에 미치는 영향)

  • Seo, Meekyung;Song, Ji-Soo;Shin, Teo Jeon;Hyun, Hong-Keun;Kim, Jung-Wook;Jang, Ki-Taeg;Lee, Sang-Hoon;Kim, Young-Jae
    • Journal of the korean academy of Pediatric Dentistry
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    • v.47 no.4
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    • pp.406-415
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    • 2020
  • Silver diamine fluoride (SDF) is an effective and efficient agent for arresting dental caries. It can be useful in treating children with behavioral or medical limitations. The purpose of this study was to evaluate the antimicrobial effect of SDF by using salivary biofilm. Pellicle-like saliva coated structure was prepared by using unstimulated saliva. For developing cariogenic biofilm, Streptococcus mutans was added to the mixture of pooled saliva and inoculated into a saliva coated glass or chamber. SDF was applied to cariogenic biofilm to evaluate the antimicrobial effect of SDF. As time passed, total bacteria and S. mutans were reduced after application of SDF (p < 0.000). Confocal laser scanning microscope also showed the increment of the ratio of dead cell. As a result of experiment using enamel and dentin of primary teeth, it was confirmed that the growth of cariogenic biofilm was inhibited when the SDF was treated (p = 0.029 each). This study showed excellent anti-microbial effect of SDF. And anti-caries effect in clinical practice can be expected.

Crocetin Induces Cytotoxicity in Colon Cancer Cells Via p53-independent Mechanisms

  • Li, Cai-Yan;Huang, Wen-Feng;Wang, Qun-Li;Wang, Fan;Cai, E.;Hu, Bing;Du, Jia-Cheng;Wang, Jing;Chen, Rong;Cai, Xiao-Jing;Feng, Jing;Li, Hui-Hui
    • Asian Pacific Journal of Cancer Prevention
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    • v.13 no.8
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    • pp.3757-3761
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    • 2012
  • Objective: Crocin has been proposed as a promising candidate for cancer chemoprevention. The purpose of this investigation was to investigate the chemopreventive action and the possible mechanisms of crocin against human colon cancer cells in vitro. Methods: Cell proliferation was examined using MTT assay and the cell cycle distribution fractions were analyzed using fow cytometric analysis after propidium iodide staining. Apoptosis was detected using theTUNEL Apoptosis Detection Kit with laser scanning confocal microscope. DNA damage was assessed using the alkaline single-cell gel electrophoresis assay, while expression levels of p53, cdk2, cyclinA and P21 were examined by Western blot analysis. Results: Treatment of SW480 cells with crocetin (0.2, 0.4, 0.8 mmol/L) for 48 h signifcantly inhibited their proliferation in a concentration-dependent manner. Crocetin (0.8 mmol/L) signifcantly induced cell cycle arrest through p53-independent mechanisms accompanied by P21 induction. Crocetin (0.8 mmol/L) caused cytotoxicity in the SW480 cells by enhancing apoptosis and decreasing DNA repair capacity in a time-dependent manner. Conclusions: This report provides evidence that crocetin is a potential anticancer agent, which may be used as a chemotherapeutic drug.