• 제목/요약/키워드: Confocal laser scanning microscopy(CLSM)

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Bacterial Dynamics of Biofilm Development During Toluene Degradation by Burkholderia vietnamiensis G4 in a Gas Phase Membrane Bioreactor

  • Kumar, Amit;Dewulf, Jo;Wiele, Tom Van De;Langenhove, Herman Van
    • Journal of Microbiology and Biotechnology
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    • 제19권9호
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    • pp.1028-1033
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    • 2009
  • In this study, the dynamics of living cells (LC) and dead cells (DC) in a laboratory-scale biofilm membrane bioreactor for waste gas treatment was examined. Toluene was used as a model pollutant. The bacterial cells were enumerated as fluoromicroscopic counts during a 140 operating day period using BacLight nucleic acid staining in combination with epifluorescence and confocal laser scanning microscopy (CSLM). Overall, five different phases could be distinguished during the biofilm development: (A) cell attachment, (B) pollutant limitation, (C) biofilm establishment and colonization, (D) colonized biofilm, and (E) biofilm erosion. The bioreactor was operated under different conditions by applying different pollutant concentrations. An optimum toluene removal of 89% was observed at a loading rate of 14.4 kg $m^{-3}d^{-1}$. A direct correlation between the biodegradation rate of the reactor and the dynamics of biofilm development could be demonstrated. This study shows the first description of biofilm development during gaseous toluene degradation in MBR.

Use of Cationic PAM as a Surface Sizing Additive to Improve Paper Properties

  • Seo, Man-Seok;Lee, Hak-Lae;Youn, Hye-Jung
    • 한국펄프종이공학회:학술대회논문집
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    • 한국펄프종이공학회 2006년도 PAN PACIFIC CONFERENCE vol.2
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    • pp.245-250
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    • 2006
  • This study was focused on the use of cationic PAM (Polyacrylamide) as a surface sizing additive to improve the surface sizing properties of paper. Effects of the ionic property, viscosity and charge density of PAM on bending stiffness of surface sized papers were investigated. Use of cationic PAM as a surface sizing additive improved bending stiffness while addition of anionic PAM did not show any effect. Increase of starch holdout with the addition of cationic PAM was attributed as a prime reason of stiffness increase. Viscosity of PAM was one of the most important factors affecting surface sizing due to its influence on the interaction between cationic PAM and oxidized starch solution. Greater improvement of bending stiffness of paper was obtained when high charged PAM was used as an additive. The order of addition was found to have significant influence on the effect of additives since it influences the formation of network structure among starch, cationic PAM, and SA (styrene acrylic acid copolymer). Investigation on the penetration of starch solution was carried out with CLSM (Confocal Laser Scanning Microscopy), and it was shown that the addition of cationic PAM to oxidized starch solution made starch molecules stay on the paper surface rather than penetrating into the paper structure because of the electrostatic interaction between negatively charged fibers and positively charged cationic PAM.

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Evaluation of Volumetric Texture Features for Computerized Cell Nuclei Grading

  • Kim, Tae-Yun;Choi, Hyun-Ju;Choi, Heung-Kook
    • 한국멀티미디어학회논문지
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    • 제11권12호
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    • pp.1635-1648
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    • 2008
  • The extraction of important features in cancer cell image analysis is a key process in grading renal cell carcinoma. In this study, we applied three-dimensional (3D) texture feature extraction methods to cell nuclei images and evaluated the validity of them for computerized cell nuclei grading. Individual images of 2,423 cell nuclei were extracted from 80 renal cell carcinomas (RCCs) using confocal laser scanning microscopy (CLSM). First, we applied the 3D texture mapping method to render the volume of entire tissue sections. Then, we determined the chromatin texture quantitatively by calculating 3D gray-level co-occurrence matrices (3D GLCM) and 3D run length matrices (3D GLRLM). Finally, to demonstrate the suitability of 3D texture features for grading, we performed a discriminant analysis. In addition, we conducted a principal component analysis to obtain optimized texture features. Automatic grading of cell nuclei using 3D texture features had an accuracy of 78.30%. Combining 3D textural and 3D morphological features improved the accuracy to 82.19%. As a comparative study, we also performed a stepwise feature selection. Using the 4 optimized features, we could obtain more improved accuracy of 84.32%. Three dimensional texture features have potential for use as fundamental elements in developing a new nuclear grading system with accurate diagnosis and predicting prognosis.

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고삼, 인삼 및 단삼 혼합물에 의한 모발의 성장과 분포에 미치는 영향 (The Effect of Hair Growth and Distribution by Sophorae Radix, Panax ginseng, Salvia miltiorrhiza BUNGE Water Extracts)

  • 황초원;심재환;김상태
    • 대한화장품학회지
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    • 제36권3호
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    • pp.215-219
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    • 2010
  • 본 연구에서는 발모의 성장결여로부터 모내기의 천연성분에 의한 침투력, 침투면적, 모발성장속도, 모발의 수 및 표면에 미치는 영향에 관하여 조사를 수행하였다. 제모를 통한 모발의 성장 속도와 모내기의 피부 침투력, 약물도달 부위를 CLSM710 공초점 현미경으로 관찰하였는데, 고삼, 단삼 및 인삼의 혼합물인 모내기를 $10\;{\mu}g/mL$ 농도로 처리한 그룹에서는 정상군과 대조군과 비교시 $33{\pm}4$$60{\pm}6$ (AFI: 인위적 형광강도)인데 반해, 약 $6782{\pm}67$ AFI (인위적 형광강도)으로 약 205배와 113배의 침투능력을 나타내었고 면적은 정상군과 대조군은 각각 약 $22{\pm}3$$44{\pm}6$ AFI인데 반해 모내기군은 약 $5602{\pm}45$ AFI였고 각각 255배와 127배 정도 침투면적의 확대되었음을 관찰하였다. 또한 confocal LSM700으로 관찰한 결과 대조군은 진피층에 존재하는 collagen IV에서 아무런 녹색형광이 없는 반면 모내기군은 collagen IV이 함유한 진피층에서 강한 녹색형광이 관찰하였는데, 이는 모내기 주성분이 피부 진피층까지 도달한 것이라 사료되고 남자 15명과 여자 5명 중 20대 4명(20 %), 30대 10명(50 %), 그리고 40대 이상 6명(30 %)의 내원환자의 탈모의 털 직경과 표면, 탈모의 수, 그리고 발모의 수를 조사하여 본 결과 평균 약 75 % 개선효과를 나타내므로 모내기의 천연성분이 발모인자의 활성화를 촉진하여 모낭 성장을 촉진시키는 기능을 유도하리라 사료된다.

Defense-Related Responses in Fruit of the Nonhost Chili Pepper against Xanthomonas axonopodis pv. glycines Infection

  • Chang, Sung Pae;Jeon, Yong Ho;Kim, Young Ho
    • The Plant Pathology Journal
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    • 제32권4호
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    • pp.311-320
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    • 2016
  • Xanthomonas axonopodis pv. glycines (Xag) is a necrotrophic bacterial pathogen of the soybean that causes bacterial pustules and is a nonhost pathogen of the chili pepper. In the current study, chili pepper fruit wound inoculated in planta with Xag 8ra formed necrotic lesions on the fruit surface and induced several structural and chemical barriers systemically in the fruit tissue. The initial defense response included programmed cell death of necrotizing and necrotized cells, which was characterized by nuclear DNA cleavage, as detected by TUNEL-confocal laser scanning microscopy (CLSM), and phosphatidylserine exposure on cell walls distal to the infection site, as detected by Annexin V FLUOS-CLSM. These two responses may facilitate cell killing and enhance transportation of cell wall materials used for cell wall thickening, respectively. The cells beneath the necrotic tissue were enlarged and divided to form periclinal cell walls, resulting in extensive formation of several parallel boundary layers at the later stages of infection, accompanying the deposition of wall fortification materials for strengthening structural defenses. These results suggest that nonhost resistance of chili pepper fruit against the nonhost necrotrophic pathogen Xag 8ra is activated systematically from the initial infection until termination of the infection cycle, resulting in complete inhibition of bacterial pathogenesis by utilizing organspecific in situ physiological events governed by the expression of genes in the plant fruit organ.

초지법에 따른 한지의 물성비교 (Comparison of Physical Properties of Hanjis Made by Different Sheet Forming Processes)

  • 최태호;조남석;최인호;정택상
    • 펄프종이기술
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    • 제33권4호
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    • pp.21-27
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    • 2001
  • Korean traditional paper (Hanji) making technology has adopted two kinds of sheet forming processes, which called "Oebal-choji": and "Ssangbal-choji". The sheet forming process of Oebal-choji is an original method developed in Korea. At first, paper stock is dipped onto the mold and flow away in the forward direction. Then, paper stock is scooped again and rhythmically rocked from side to side, this work is repeated several times. Through this operation the fibers intertwine and paper layers are formed. Ssangbal-choji is almost same as the Nagashizuki, which used in Japan. In this method, paper stock is scooped onto the mold and rhythmically rocked backwards and forwards several times, the water drains slowly through the bamboo screen and then sheet is formed. Tamezuki method is used in Japan and China. This is a method in which the mold is dipped into the paper stock once and left to drain. In the Ssangbal-choji and Nagashizuki methods, the most of excess solution is cast out while in the Tamezuki all of it is allowed to drain through the mold. This study was carried out to investigate the physical properties of the Hanjis that were made by Oebal-choji, Ssangbal-choji, Nagashizuki, and Tamezuki sheet forming processes. The results were follows; Physical properties of the Oebal-choji Hanji were better than those of Ssangbal-choji, Nagashizuki, and Tamezuki. Oebal-choji Hanji made little difference of paper strength between MD and CD, but Ssangbal-chjo and Nagashizuki Hanjis made wide difference. And there are no difference of paper strength between MD and CD on the Tamezuki Hanji. On the confocal laser scanning microscopy (CLSM) observation of the Hanjis, Oebal-choji made well crossed fiber orientation than those of other forming processes.r forming processes.

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Tuberostemonine에 의한 Staphylococcus aureus의 생물막 억제 효과 (Inhibitory effects of tuberostemonine on Staphylococcus aureus biofilm)

  • 염수진;김승민;권준혁;정희곤
    • 한국식품과학회지
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    • 제54권2호
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    • pp.241-246
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    • 2022
  • 본 연구는 S. aureus에 대한 tuberostemonine의 항균 및 항생물막 효과에 대하여 확인하였다. S. aureus에 대한 tuberostemonine의 생장 저해 효과가 없음을 확인하였으나, crystal violet 염색법과 CLSM 이미지 측정을 통해 tuberostemonine이 유의한 S. aureus 항생물막 효과 가지는 것을 알 수 있었다. S. aureus의 생물막 형성과 분해 관련 유전자인 icaA와 agrA의 발현은 tuberostemonine를 처리하였을 때 유의미하게 각각 감소 또는 증가하는 것으로 나타났다. 따라서 본 연구에서 생물막 형성 저해 및 분해 효과가 확인된 천연화합물인 tuberostemonine은 S. aureus의 내성 발생 위험이 적은 새로운 항생물막제제로서 사용가능 할 것으로 사료된다.

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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    • 제47권4호
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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.

EDTA Surface Capped Water-Dispersible ZnSe and ZnS:Mn Nanocrystals

  • Lee, Jae-Woog;Lee, Sang-Min;Huh, Young-Duk;Hwang, Cheong-Soo
    • Bulletin of the Korean Chemical Society
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    • 제31권7호
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    • pp.1997-2002
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    • 2010
  • ZnSe and ZnS:Mn nanocrystals were synthesized via the thermal decomposition of their corresponding organometallic precursors in a hot coordinating solvent (TOP/TOPO) mixture. The organic surface capping agents were substituted with EDTA molecules to impart hydrophilic surface properties to the resulting nanocrystals. The optical properties of the water-dispersible nanocrystals were analyzed by UV-visible and room temperature solution photoluminescence (PL) spectroscopy. The powders were characterized by X-ray diffraction (XRD), high resolution transmission electron microscopy (HR-TEM), and confocal laser scanning microscopy (CLSM). The solution PL spectra revealed emission peaks at 390 (ZnSe-EDTA) and 597 (ZnS:Mn-EDTA) nm with PL efficiencies of 4.0 (former) and 2.4% (latter), respectively. Two-photon spectra were obtained by fixing the excitation light source wavelengths at 616 nm (ZnSe-EDTA) and 560 nm (ZnS:Mn-EDTA). The emission peaks appeared at the same positions to that of the PL spectra but with lower peak intensity. In addition, the morphology and sizes of the nanocrystals were estimated from the corresponding HR-TEM images. The measured average particle sizes were 5.4 nm (ZnSe-EDTA) with a standard deviation of 1.2 nm, and 4.7 nm (ZnS:Mn-EDTA) with a standard deviation of 0.8 nm, respectively.

Antifungal effects of synthetic human β-defensin 3-C15 peptide

  • Lim, Sang-Min;Ahn, Ki-Bum;Kim, Christine;Kum, Jong-Won;Perinpanayagam, Hiran;Gu, Yu;Yoo, Yeon-Jee;Chang, Seok Woo;Han, Seung Hyun;Shon, Won-Jun;Lee, Woocheol;Baek, Seung-Ho;Zhu, Qiang;Kum, Kee-Yeon
    • Restorative Dentistry and Endodontics
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    • 제41권2호
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    • pp.91-97
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    • 2016
  • Objectives: The purpose of this ex vivo study was to compare the antifungal activity of a synthetic peptide consisting of 15 amino acids at the C-terminus of human ${\beta}$-defensin 3 (HBD3-C15) with calcium hydroxide (CH) and Nystatin (Nys) against Candida albicans (C. albicans) biofilm. Materials and Methods: C. albicans were grown on cover glass bottom dishes or human dentin disks for 48 hr, and then treated with HBD3-C15 (0, 12.5, 25, 50, 100, 150, 200, and $300{\mu}g/mL$), CH ($100{\mu}g/mL$), and Nys ($20{\mu}g/mL$) for 7 days at $37^{\circ}C$. On cover glass, live and dead cells in the biomass were measured by the FilmTracer Biofilm viability assay, and observed by confocal laser scanning microscopy (CLSM). On dentin, normal, diminished and ruptured cells were observed by field-emission scanning electron microscopy (FE-SEM). The results were subjected to a two-tailed t-test, a one way analysis variance and a post hoc test at a significance level of p = 0.05. Results: C. albicans survival on dentin was inhibited by HBD3-C15 in a dose-dependent manner. There were fewer aggregations of C. albicans in the groups of Nys and HBD3-C15 (${\geq}100{\mu}g/mL$). CLSM showed C. albicans survival was reduced by HBD3-C15 in a dose dependent manner. Nys and HBD3-C15 (${\geq}100{\mu}g/mL$) showed significant fungicidal activity compared to CH group (p < 0.05). Conclusions: Synthetic HBD3-C15 peptide (${\geq}100{\mu}g/mL$) and Nys exhibited significantly higher antifungal activity than CH against C. albicans by inhibiting cell survival and biofilm.