• 제목/요약/키워드: CLSM Confocal laser scanning microscope

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Application of the Homogenization Analysis to Calculation of a Permeability Coefficient (투수계수 산정을 위한 균질화 해석법의 적응)

  • 채병곤
    • Journal of Soil and Groundwater Environment
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    • v.9 no.1
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    • pp.79-86
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    • 2004
  • Hydraulic conductivity along rock fracture is mainly dependent on fracture geometries such as orientation, aperture, roughness and connectivity. Therefore, it needs to consider fracture geometries sufficiently on a fracture model for a numerical analysis to calculate permeability coefficient in a fracture. This study performed new type of numerical analysis using a homogenization analysis method to calculate permeability coefficient accurately along single fractures with several fracture models that were considered fracture geometries as much as possible. First of all, fracture roughness and aperture variation due to normal stress applied on a fracture were directly measured under a confocal laser scaning microscope (CLSM). The acquired geometric data were used as input data to construct fracture models for the homogenization analysis (HA). Using the constructed fracture models, the homogenization analysis method can compute permeability coefficient with consideration of material properties both in microscale and in macroscale. The HA is a new type of perturbation theory developed to characterize the behavior of a micro inhomogeneous material with a periodic microstructure. It calculates micro scale permeability coefficient at homogeneous microscale, and then, computes a homogenized permeability coefficient (C-permeability coefficient) at macro scale. Therefore, it is possible to analyze accurate characteristics of permeability reflected with local effect of facture geometry. Several computations of the HA were conducted to prove validity of the HA results compared with the empirical equations of permeability in the previous studies using the constructed 2-D fracture models. The model can be classified into a parallel plate model that has fracture roughness and identical aperture along a fracture. According to the computation results, the conventional C-permeability coefficients have values in the range of the same order or difference of one order from the permeability coefficients calculated by an empirical equation. It means that the HA result is valid to calculate permeability coefficient along a fracture. However, it should be noted that C-permeability coefficient is more accurate result than the preexisting equations of permeability calculation, because the HA considers permeability characteristics of locally inhomogeneous fracture geometries and material properties both in microscale and macroscale.

The Study of Restoration Technique of Wax-Treated Volume for the Annals of the Joseon Dynasty (III) -Evaluation of Durability of Korean Traditional Paper using Moist-heat Aging Treatment- (조선왕조실록 밀랍본 복원기술 연구(제3보) -습열열화처리를 이용한 복원용 한지의 내구성 평가-)

  • Jeong, Seon-Hwa;Jeong, Sun-Young;Seo, Jin-Ho;Jeong, So-Young
    • Journal of Korea Technical Association of The Pulp and Paper Industry
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    • v.45 no.5
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    • pp.49-55
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    • 2013
  • To explore the paper materials for restoration of the Annals of the Joseon Dyansty, durability of the three type of the traditional Korean Papers were estimated in this study, through moist heat artificial aging test. Three types(D, F, and G) which showed the best preservation performance in dry heat and UV treatment in the previous study were selected and artificial accelerated aging treatment with moist-heat process was conducted; the viscosity change rate was D>G>F; folding endurance G>D>F; $L^*$ value F>D>G; $a^*$ and $b^*$ change rate D>G>F; brightness decrease rate D>G>F, suggesting paper F showed the least change rate in physical/optical properties. Also the CLSM image observation showed fair coherence among fibers and confirmed paper mulberry. And in FDI extraction from each sample, paper F showed the highest value. Overall, paper F (traditional glossy paper) showed the highest stability against thermal treatment. It confirms that paper F is suitable as restoration paper for tributary remains including the annals of the Joseon Dynasty for its steady strength/viscosity decrease rate and color change rate.

Effect of NaCl on Biofilm Formation of the Isolate from Staphylococcus aureus Outbreak Linked to Ham

  • Lee, Soomin;Choi, Kyoung-Hee;Yoon, Yohan
    • Food Science of Animal Resources
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    • v.34 no.2
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    • pp.257-261
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    • 2014
  • The objective of this study was to evaluate the effects of NaCl on the biofilm formations of the isolate from Staphylococcus aureus outbreaks linked to ham. The S. aureus ATCC13565 isolated from ham was exposed to NaCl concentrations of 0%, 2%, 4%, and 6% supplemented in tryptic soy broth (TSB) for 24 h at $35^{\circ}C$, followed by plating 0.1 mL of the culture on tryptic soy agar containing 0%, 2%, 4%, and 6% NaCl, respectively. After incubating at $35^{\circ}C$ for 24 h, the colonies on the plates were collected and diluted to $OD_{600}$ = 0.1. The diluents of S. aureus were incubated on a 96-well flat bottom plate containing TSB plus the appropriate NaCl concentrations, and the biofilm formation was quantified by crystal violet staining after being incubated at $35^{\circ}C$ for 9 h. Confocal laser scanning microscope (CLSM) was also used for visualizing the biofilm formation of S. aureus at NaCl concentrations of 0%, 2%, 4%, and 6%. The transcriptional analysis of biofilm-related genes, such as icaA, atl, clfA, fnbA, sarA, and rbf, was conducted by quantitative real-time PCR. Crystal violet staining and CLSM showed that the biofilm formations of S. aureus increased (p<0.05) along with the NaCl concentrations. Moreover, the expression of the icaA genes was higher at the NaCl concentrations of 4% and 6% as compared with 0% of NaCl by approximately 9-folds and 20-folds, respectively. These results indicated that the NaCl formulated in processed food may increase the biofilm formations of S. aureus by increasing the icaA gene expressions.

Preliminary study on the effect of inflamed TMJ synovial fluid on the intracellular calcium concentration and differential expression of iNOS and COX-2 in human immortalized chondrocyte C28/I2

  • Choi, Eun-Ah;Lee, Dong-Geun;Chae, Chang-Hoon;Chang, Young-Il;Park, Young-Ju;Kim, Young-Kyun
    • Journal of the Korean Association of Oral and Maxillofacial Surgeons
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    • v.32 no.1
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    • pp.36-41
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    • 2006
  • Objective. The objective of this study was to examine the hypothesis that inflammatory synovial fluid from TMJ internal derangement initiates a transient increase in intracellular calcium concentration ([$Ca^{2+}$]i) in chondrocytes and the induced Ca2+ signaling affects iNOS/COX-2 gene expression patterns following exposure to inflamed synovial fluid. Materials and Methods. Two female adult patients with symptoms of TMD who agreed to participate in the study were selected for this study. Immortalized human juvenile costal chondrocyte C-28/I2 was grown to 80% confluency and synovial fluids from two patients were added respectively to culture media for 24 hours at the concentration of 100ng/10ml. Confocal laser scanning microscope (CLSM) was used to examine changes of intracellular calcium concentration ([$Ca^{2+}$]i). RT-PCR was performed to identify the expression profile of IL-1${\alpha}$, iNOS, COX-2. Results. Increased [$Ca^{2+}$]i was observed in chondrocytes subjected to inflamed synovial fluid compared to control cultures and in respective cultures exposed to inflamed synovial fluids from each patient, IL-1${\beta}$, COX-2 mRNA were detected. However, in neither case iNOS mRNA was expressed. IL-1${\alpha}$, COX-2, and iNOS mRNA were expressed in control culture. Conclusion. Our results show that immortalized chondrocytes cultured with inflamed synovial fluids from patients diagnosed as disc displacement without reduction and limitation in mouth opening showed increased calcium concentration and expression of COX-2 while inhibiting the production of iNOS, which in turn could adversely affect the chondrocytes in at least short term by hindering physiologic role of NO against inflammatory cascades. These findings suggest that inflamed synovial fluid may differentially regulate the transcriptomes of relevant inflammatory mediators, especially iNOS/COX-2 axis in chondrocytes through adjusting calcium transients.

The Surface Characteristic Changes of Hydroxyapatite Coated Ti Disc When Immersed in NaCl Solution (NaCl 수용액에 담근 Hydroxyapatite 코팅된 타이타늄 시편의 표면 변화)

  • Baek, Yeon-Wha;Kim, Myung-Joo;Kwon, Ho-Beom;Lim, Young-Jun
    • Journal of Dental Rehabilitation and Applied Science
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    • v.28 no.4
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    • pp.339-347
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    • 2012
  • In previous studies, methods for enhancing cellular response on the Hydroxyapatite coated implant surface were described. In this study, the changes of surface characteristics such as surface roughness, contact angle, surface energy and surface morphology were observed when Hydroxyapatite coated Ti discs were immersed in NaCl solution for various time. Hydroxyapatite coated Ti discs were immersed in 0.9% NaCl solution for 7, 14 and 21 days at $37^{\circ}C$. The control group comprises dry identical discs not immersed in a solution. (n=3) All discs were dried in air completely and the surface roughness was measured using confocal laser scanning microscopy(CLSM). Static contact angle was recorded by video contact angle analyzer after dropping distilled water on the surface. The surface energy was calculated from contact angles of the three liquids. Surface was observed using a field emission-scanning electron microscope(FE-SEM). As a result, the surface roughness of immersed Hydroxyapatite coated Ti discs increased significantly and the contact angle decreased comparing with control group discs. The surface energy of immersed discs increased except for discs immersed for 14 days.

Red-Colored Phenomena and Morphochemical Characteristics of Red-Colored Substances in Ginseng Roots (Panax ginseng C.A. Meyer) (인삼 적변현상과 적변물질의 형태-화학적 특성)

  • 윤길영;양덕조
    • Journal of Ginseng Research
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    • v.24 no.3
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    • pp.107-112
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    • 2000
  • One of the physiologically important ginseng diseases is red-colored phenomena (RCP) that is caused by accumulation of red-colored substances on the epidermis of ginseng roots. Although RCP severely deteriorates the quality of ginseng products, there has been little information on what red-colored substance is and how RCP occurs. Therefore, the heavy losses of cultivators and ginseng industry are suffering by RCP, For this reason, we have investigated with the morphochernical characteristics of RCP to find out main cause of it. The red-colored substances (RS) on the epidermis of red-colored ginseng (RCG) were examined using inverted light microscope, confocal laser scanning microscope (CLSM)and furier transform infrared (FT/IR) spectrometer. Red brown substances were accumulated in the cell wall of the epidermis from early stage to late stage of RCC. Especially, cell wall of the late stage of RCG was covered with the sub-stances with 80~ 130 fm thick. Therefore, the cell wall of RCG cannot protect the ginseng root cells from the mechanical damages, bacteria and fungi. To analyse red substances of roots, RS were isolated from epidermis of RCG and extracted using various solvents. RS is strongly insoluble but it was bleached by oxidizing agents including 12% (v/v) NaOCl. Therefore, RS was Presumed to make up of high chelation power. The proriles of FT/IR spectra or both healthy ginseng (HEG) and RCG showed a significant difference at two wavelength,2857 cm$\^$-1/(C-H) and 1032 cm$\^$-1/(S=O), respectively. Furthermore, absorption peak of 2857cm$\^$-l/ appears on the only epidermis of RCG. The other peak is shown lower absorption rate on the epidermis of RCG than that of healthy ginseng. Also, FT/IR spectra of the mixture of carboxym-ethylcellulose (CMC) and iron (Fe$\^$3+/) were very similar to RCG spectrum profiles. One of a interesting fact is that the contents of phenolic compounds at the epidermis of healthy ginseng were highest. The results of these experiments sup-port the RCP was closely related with the chemical interaction between inorganic elements (Fe) of rhizosphere and organic matters (cellulose, cellobiose, cell sap, etc.) of ginseng roots.

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EFFECTS OF DENTIN SURFACE WETNESS OR DESICCATION AFTER ACID ETCHING ON DENTIN BONDING (산부식후 상아질 표면의 습윤 또는 건조가 상아질 결합에 미치는 영향)

  • Yang, Won-Kyung;Kwon, Hyuk-Choon;Son, Ho-Hyun
    • Restorative Dentistry and Endodontics
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    • v.25 no.2
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    • pp.243-253
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    • 2000
  • The purpose of this in vitro study was to evaluate dentin bonding by two different dentin bonding systems(DBS) using acetone based primer or adhesive [All Bond 2(AB2), One Step(OS)] when they were applied by wet or dry bonding technique. Morphology of resin-dentin interface and hybrid layer thickness(HLT) were investigated using Confocal Laser Scanning Microscope(CLSM) and compared to shear bond strength(SBS). 72 extracted sound human molars were randomly divided into 4 groups of 18 teeth each - Group 1.(AW); AB2 by wet bonding. Group 2(AD); AB2 by dry bonding. Group 3.(OW); OS by wet bonding, Group 4.(OD); OS by dry bonding. In 6 teeth of each group, notch-shaped class V cavities(depth 2mm) were prepared on buccal and lingual surface at the cementoenamel juction(12 cavities per group). To obtain color contrast in CLSM observation, bonding resins of each DBS were mixed with rhodamine B and primer of AB2 was mixed with sodium fluorescein. Prepared teeth of each group were treated with AB2, OS, respectively according to the manufacturer's instructions except for dentin surface moisture treatment after acid etching. In group 1 and 3, after acid etching, excess water was removed with wet tissue(Kimwipes), leaving consistently shiny, visibly hydrated dentin surface. In group 2 and 4, dentin surface was dried for 10 seconds at 1 inch distance. The treated teeth were then packed with composite resin(${\AE}$litefil) and light-cured. 12 microscopic samples($60{\sim}80{\mu}m$ thickness) of each group were obtained after longitudinal section and grinding(Exakt cutting and grinding system). Morphological investigation of resin-dentin interface and HLT measurement using CLSM were done. For measurement of SBS, remaining 12 teeth of each group were flattened occlusally to remove all enamel and grinded to 500 grit SiC(Pedemet Specimen Preparation Equipment). After applying DBS on the exposed dentin surface, composite resin was applied in the shape of cylinder, which has 5mm diameter, 1.5mm thickness, and light cured. SBS was measured using Instron with a crosshead speed of 0.5mm/min. It was concluded as follows, 1. HLT of AW(mean: $2.59{\mu}m$) was thicker than any other group, and followed by AD, OW, OD in descending order(mean; 2.37, 2.28, $1.92{\mu}m$). Only OD had statistically significant differences(p<0.05) to AW and AD. 2. There were intimate contact of resin and dentin at the interface in wet bonding groups, but gaps or irregular interfaces were observed in dry bonding groups. 3. The length, diameter, density of resin tags were various even in the same group without significant differences between groups and lots of adhesive lateral branches were observed. 4. There were no statistically significant difference of SBS between AB2 and OS, but SBS of wet bonding groups were significantly higher(p<0.05) than dry bonding groups. 5. There were no consistent relationships between HLT and SBS.

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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.

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.