• Title/Summary/Keyword: Confocal laser scanning microscopy

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Antifungal Mechanism of Antifungal Peptide Derived from Cecropin A(1-8)- Melittin(1-12) Hybrid against Aspergillus fumigatus

  • Lee, Dong-Gun;Jin, Zhe-Zhu;Maeng, Cheol-Young;Shin, Song-Yub;Seo, Moo-Yeol;Kim, Kil-Lyong;Hahm, Kyung-Soo
    • Journal of Microbiology and Biotechnology
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    • v.9 no.2
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    • pp.168-172
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    • 1999
  • The antifungal mechanism of the antifungal peptide against Aspergillus fumigatus, $K^{18,19}$-CA(l-8)-ME(l-12), derived from cecropin A(l-8)-melittin(l-12) was investigated by confocal laser scanning microscopy, cell wall regeneration, ATPase activity inhibition, and released potassium ion. By confocal laser scanning microscopy, $K^{18,19}$-CA(l-8)-ME(l-12) was detected on the surface of A. fumigatus, while cecropin A used as a negative control peptide was not detected. The protoplast of A. fumigatus treated with$K^{18,19}$-CA(1-8)-ME(1-12) failed to regenerate the fungal cell walls. Compared with cecropin A, the amount of potassium ion released by $K^{18,19}$-CA(l-8)-ME(l-12) was increased. Furthermore, $K^{18,19}$-CA(l-8)-ME(l-12) inhibited the ATPase activity on the plasma membrane. These results suggested that $K^{18,19}$-CA(l-8)-ME(1-12) acts on the plasma membrane of A. fumigatus and its antifungal action is due to the ion channel or pore formation on the plasma membrane.

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Sheet Formation Properties of Morus Hanji (뽕나무 한지의 지합특성)

  • Yi, Il-Roh;Cho, Nam-Seok
    • Journal of Korea Technical Association of The Pulp and Paper Industry
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    • v.38 no.3 s.116
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    • pp.47-60
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    • 2006
  • This study was carried out to investigate the sheet formation properties of Morus Hanjis, made of bast and whole stalk pulps by different pulping methods, such as alkali, alkali-peroxide and sulfomethylated pulping. Two species of Morus, M. alba and M. lhou, were used. Effect of morphological properties of pulp stocks on the sheet formation and its gray levels based on optical property were evaluated using an Image analyzer. In addition, the effect of fiber distribution index(FDI) which was calculated from tile data of Confocal laser scanning microscopy(CLSM) on the sheet formation and optical properties of Morus Hanji were also discussed. On the sheet formation, Hanji from whole stalk pulp was superior than that of bast pulp. The more the sheet formation improved, the more paper opacity decreased. In the aspect of Hanji's surface characteristics analyzed by an Image analyzer, the average gray level and its standard deviation of Hanji from the whole stalk pulp were rather lower than those of bast pulp because of better sheet formation of the former. However, high brightness Hanji showed high value of gray level. The sheet formation and paper opacity were increased with the decrease of standard deviation of gray level. From these results, gray level measurement could be used to predict the paper opacity as well as sheet formation. Sheet formation of whole stalk Hanji and its FDI measured by CLSM were higher than those of bast fibers. In conclusion, the sheet formation and opacity of Hanji could be evaluated by standard deviation value of Hanji's gray level using an Image analyzer and by fiber distribution index using CLSM.

Effects of Nutritional and Environmental Conditions on Planktonic Growth and Biofilm Formation of Citrobacter werkmanii BF-6

  • Zhou, Gang;Li, Long-Jie;Shi, Qing-Shan;Ouyang, You-Sheng;Chen, Yi-Ben;Hu, Wen-Feng
    • Journal of Microbiology and Biotechnology
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    • v.23 no.12
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    • pp.1673-1682
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    • 2013
  • Citrobacter sp. is a cause of significant opportunistic nosocomial infection and is frequently found in human and animal feces, soil, and sewage water, and even in industrial waste or putrefaction. Biofilm formation is an important virulence trait of Citrobacter sp. pathogens but the process and characteristics of this formation are unclear. Therefore, we employed in vitro assays to study the nutritional and environmental parameters that might influence biofilm formation of C. werkmanii BF-6 using 96-well microtiter plates. In addition, we detected the relative transcript levels of biofilm formation genes by RT-PCR. Our results indicated that the capacity of C. werkmanii BF-6 to form biofilms was affected by culture temperature, media, time, pH, and the osmotic agents glucose, sucrose, NaCl, and KCl. Confocal laser scanning microscopy results illustrated that the structure of biofilms and extracellular polysaccharide was influenced by 100 mM NaCl or 100 mM KCl. In addition, nine biofilm formation genes (bsmA, bssR, bssS, csgD, csgE, csgF, mrkA, mrkB, and mrkE) were found to contribute to planktonic and biofilm growth. Our data suggest that biofilm formation by C. werkmanii BF-6 is affected by nutritional and environmental factors, which could pave the way to the prevention and elimination of biofilm formation using proper strategies.

Improved dentin disinfection by combining different-geometry rotary nickel-titanium files in preparing root canals

  • Bedier, Marwa M.;Hashem, Ahmed Abdel Rahman;Hassan, Yosra M.
    • Restorative Dentistry and Endodontics
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    • v.43 no.4
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    • pp.46.1-46.10
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    • 2018
  • Objectives: This study was to evaluate the antibacterial effect of different instrumentation and irrigation techniques using confocal laser scanning microscopy (CLSM) after root canal inoculation with Enterococcus faecalis (E. faecalis). Materials and Methods: Mesiobuccal and mesiolingual canals of extracted mandibular molars were apically enlarged up to a size 25 hand K-file, then autoclaved and inoculated with E. faecalis. The samples were randomly divided into 4 main groups according to the system of instrumentation and irrigation: an XP-endo Shaper (XPS) combined with conventional irrigation (XPS/C) or an XP-endo Finisher (XPF) (XPS/XPF), and iRaCe combined with conventional irrigation (iRaCe/C) or combined with an XPF (iRaCe/XPF). A middle-third sample was taken from each group, and then the bacterial reduction was evaluated using CLSM at a depth of $50{\mu}m$ inside the dentinal tubules. The ratio of red fluorescence (dead cells) to green-and-red fluorescence (live and dead cells) represented the percentage of bacterial reduction. The data were then statistically analyzed using the Kruskal-Wallis test for comparisons across the groups and the Dunn test was used for pairwise comparisons. Results: The instrumentation and irrigation techniques had a significant effect on bacterial reduction (p < 0.05). The iRaCe/XPF group showed the strongest effect, followed by the XPS/XPF and XPS/C group, while the iRaCe/C group had the weakest effect. Conclusions: Combining iRaCe with XPF improved its bacterial reduction effect, while combining XPS with XPF did not yield a significant improvement in its ability to reduce bacteria at a depth of $50{\mu}m$ in the dentinal tubules.

COMPARATIVE STUDY ON THE EFFICACY OF DIGITAL IMAGING FIBER-OPTIC TRANS-ILLUMINATION AND LASER FLUORESCENCE IN MONITORING THE REMINERALIZATION PROCESS OF INCIPIENT SMOOTH SURFACE ENAMEL LESIONS (Digital Imaging Fiber-Optic Trans-Illumination과 Laser Fluorescence를 이용한 평활면 초기우식증의 재광화에 관한 비교 연구)

  • Hwang, Kyu-Seon;Yoo, Seung-Hoon;Kim, Jong-Soo
    • Journal of the korean academy of Pediatric Dentistry
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    • v.34 no.2
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    • pp.183-191
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    • 2007
  • Through out the world dental caries seems to be decreased as it is difficult to make an accurate diagnosis for dental caries. The traditional diagnostic method which is probing and x-ray taking has many limitations to diagnose the early caries, so there were recommendations for the needs of new equipments such as laser fluorescence(LF), digital imaging fiber-optic trans-illumination(DIFOTI), and quantitative light fluorescence (QLF) which were developed from various study results. Also confocal laser scanning microscopy(CLSM) and ultrasonics are used for research progression. This study is to evaluate whether it is possible to monitor accurately for remineralization amount of enamel surface early caries using DIFOTI or LF After inducing artificial caries to bovine teeth to 10 participants remineralization was enhanced by 0 ppm and 500 ppm fluoride mouth rinse solution for 3 weeks. Then they were cross sectioned and analyzed using gold standard of the lesion depth measured by CLSM. The following results were obtained: 1. The measured percentage of light intensity(luminosity ratio) by DIFOTI increased with remineralization period, and showed significant reverse correlation with lesion depth measured by CLSM (p<0.01). 2. The measurement of laser fluorescence increased with remineralization period, and showed significant correlation with lesion depth measured by CLSM (p<0.01). 3. To the result for CLSM, 500 ppm fluoride mouth rinse group showed rapid rate for decreased tendency of lesion depth than 0 ppm fluoride mouth rinse group. In conclusion DIFOTI system was used to measure accurately for the remineralization amount of early surface caries, it is a very useful equipment to detect precisely the changes for early enamel caries remineralization during treatments.

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Detection of Mitochondrial Reactive Oxygen Species in Living Rat Trigeminal Caudal Neurons

  • Lee, Hae In;Chun, Sang Woo
    • International Journal of Oral Biology
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    • v.40 no.2
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    • pp.103-109
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    • 2015
  • Growing evidence suggests that mitochondrial reactive oxygen species (ROS) are involved in various pain states. This study was performed to investigate whether ROS-induced changes in neuronal excitability in trigeminal subnucleus caudalis are related to ROS generation in mitochondria. Confocal scanning laser microscopy was used to measure ROS-induced fluorescence intensity in live rat trigeminal caudalis slices. The ROS level increased during the perfusion of malate, a mitochondrial substrate, after loading of 2',7'-dichlorofluorescin diacetate ($H_2DCF-DA$), an indicator of the intracellular ROS; the ROS level recovered to the control condition after washout. When pre-treated with phenyl N-tert-butylnitrone (PBN) and 4-hydroxy-2,2,6,6-tetramethylpiperidene-1-oxyl (TEMPOL), malate-induced increase of ROS level was suppressed. To identify the direct relation between elevated ROS levels and mitochondria, we applied the malate after double-loading of $H_2DCF-DA$ and chloromethyl-X-rosamine (CMXRos; MitoTracker Red), which is a mitochondria-specific fluorescent probe. As a result, increase of both intracellular ROS and mitochondrial ROS were observed simultaneously. This study demonstrated that elevated ROS in trigeminal subnucleus caudalis neuron can be induced through mitochondrial-ROS pathway, primarily by the leakage of ROS from the mitochondrial electron transport chain.

CLSM [Confocal Laser Scanning Microscope] Observation of the Surface Roughness of Pressurized Rock Samples During Freeze/Thaw Cycling

  • Kim, Hye-jin;Choi, Junghae;Chae, Byung-gon;Kim, Gyo-won
    • The Journal of Engineering Geology
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    • v.25 no.2
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    • pp.165-178
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    • 2015
  • Physical and chemical weathering degrades rock, affecting its structural properties and thus the stability of stone buildings or other structures. Confocal laser scan microscopy (CLSM) is used here to observe temporal changes in the surface roughness of rock samples under simulated accelerated weathering. Samples were pressurized to 50, 55, or 70 MPa using a pressure frame, and subjected to freeze/thaw cycling controlled by a thermostat. The temperature was cycled from -20℃ to 40℃ and back. After each 20 cycles, CLSM was used to assess the change in surface roughness, and roughness factors were calculated to quantify the progression of the surface condition over time. Variations in cross-section line-roughness parameters and surface-roughness parameters were analyzed for specific parts of the sample surfaces at 5× and 50× magnification. The result reveals that the highest and lowest values of the roughness factors are changed according to elapsed time. Freezing/thawing at high pressure caused larger changes in the roughness factor than at low pressure.

Effect of Photothermal Therapy with Indocyanine Green in Multispecies Biofilm (Indocyanine Green을 이용한 광열 치료의 다종 우식원성 바이오필름에 대한 효과)

  • Kim, Myunghwan;Park, Howon;Lee, Juhyun;Seo, Hyunwoo;Lee, Siyoung
    • Journal of the korean academy of Pediatric Dentistry
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    • v.48 no.1
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    • pp.21-30
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    • 2021
  • The purpose of this study is to investigate the antibacterial effects of indocyanine green (ICG) and near-infrared diode lasers on multispecies biofilms. Multispecies biofilms of Streptococcus mutans, Lactobacillus casei and Candida albicans were treated with different irradiation time using photosensitizer ICG and 808 nm near-infrared diode laser. Colony forming unit (CFU) was measured, and qualitative evaluation of biofilm was performed with confocal laser scanning microscopy (CLSM). Temperature measurement was conducted to evaluate photothermal effect. In the groups using ICG and diode laser, reduction in CFU was statistically significant, but the difference in antibacterial effect on L. casei and C. albicans with irradiation time was not significant, and similar results were confirmed with CLSM. Groups with ICG and diode laser showed higher temperature elevation than groups without ICG, and results of measured temperature were similar to the range of hyperthermia. In conclusion, ICG and near-infrared diode laser showed antibacterial effects on multispecies biofilms, but studies on protocol are necessary for clinical application.

SURFACE ROUGHNESS OF EXPERIMENTAL COMPOSITE RESINS USING CONFOCAL LASER SCANNING MICROSCOPE (공초점 레이저 주사 현미경을 이용한 실험적 레진의 표면 조도에 대한 연구)

  • Bae, J.H.;Lee, M.A.;Cho, B.H.
    • Restorative Dentistry and Endodontics
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    • v.33 no.1
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    • pp.1-8
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    • 2008
  • The purpose of this study was to evaluate the effect of a new resin monomer, filler size and polishing technique on the surface roughness of composite resin restorations using confocal laser scanning microscopy. By adding new methoxylated Bis-GMA (Bis-M-GMA, 2,2-bis[4-(2-methoxy-3-methacryloyloxy propoxy) phenyl] propane) having low viscosity, the content of TEGDMA might be decreased. Three experimental composite resins were made: EX1 (Bis-M-GMA/TEGDMA = 95/5 wt%, 40 nm nanofillers); EX2 (Bis-M-GMA/TEGDMA = 95/5 wt%, 20 nm nanofillers); EX3 (Bis-GMA/TEGDMA = 70/30 wt%, 40 nm nanofillers). Filtek Z250 was used as a reference. Nine specimens (6 mm in diameter and 2 mm in thickness) for each experimental composite resin and Filtek Z250 were fabricated in a teflon mold and assigned to three groups. In Mylar strip group, specimens were left undisturbed. In Sof-lex group, specimens were ground with #1000 SiC paper and polished with Sof-lex discs. In DiaPolisher group, specimens were ground with #1000 SiC paper and polished with DiaPolisher polishing points. The Ra (Average roughness), Rq (Root mean square roughness), Rv (Valley roughness), Rp (Peak roughness), Rc (2D roughness) and Sc (3D roughness) values were determined using confocal laser scanning microscopy. The data were statistically analyzed by Two-way ANOVA and Tukey multiple comparisons test (p = 0.05). The type of composite resin and polishing technique significantly affected the surface roughness of the composite resin restorations (p < 0.001). EX3 showed the smoothest surface compared to the other composite resins (p < 0.05). Mylar strip resulted in smoother surface than other polishing techniques (p < 0.05). Bis-M-GMA. a new resin monomer having low viscosity, might reduce the amount of diluent, but showed adverse effect on the surface roughness of composite resin restorations.

Morphology of Silk Fibroin/Poly(vinyl alcohol) Blend Film (견피브로인/Poly(vinyl alcohol) 브렌드 필름의 형태학적 구조)

  • 엄인철;박영환
    • Journal of Sericultural and Entomological Science
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    • v.40 no.2
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    • pp.169-175
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    • 1998
  • The morphology of silk fibroin/poly(vinyl alcohol)(PVA)blend films was investigated using optical microscopy and confocal laser scanning microscopy. The effects of blend ratio and molecular weight of silk fibroin and PVA on phase separation were studied. Macro-phase separation occurred for the silk fibroin-rich/poor region whereas micro-phase separation took place for the dispersed/continuous phase, In spite of differences in molecular weight and blend ratio, it is observed that the dispersed phase and continuous one are composed of silk fibroin and PVA component, respectively. As the molecular weight of silk fibroin and silk fibroin content in blend ratio are decreased, the compatibility of blend is increased due to the reduction of micro-phase separation.

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