• Title/Summary/Keyword: confocal laser scanning microscopy (CLSM)

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The Cosmeceutical Property of Antioxidant Astaxanthin is Enhanced by Encapsulation Using Glyceryl Based New Vesicle (글리세릴 베이스의 신규베지클 이용 캡슐화를 통한 항산화성 아스타잔틴의 성질 강화)

  • Kim, Dong Myung;Hong, Weon Ki;Kong, Soo Sung;Lee, Chung Hyun
    • Journal of the Society of Cosmetic Scientists of Korea
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    • v.40 no.3
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    • pp.247-257
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    • 2014
  • Oil-in-water nanoemulsions of astaxanthin prepared by new vesicle, glyceryl citrate/ lactate/ linoleate/ oleate, were evaluated thoroughly in terms of cosmeceutical properties such as antioxidant effect, cell viability, influence of protein related enzyme, skin penetration, skin hydration and elasticity. Antioxidant effect and cell viability of nanoemulsion of astaxanthin were evaluated by DPPH and MTT assay. Also other properties of nanoemulsions of astaxanthin were measured by proteome analysis using 2D-PAGE, confocal laser scanning microscope and in-vivo test. We were able to find that the nanoemulsion of astaxanthin is good at scavenging of radical and inhibits the degradation of dermal extracellular matrix with the down-regulation of MMPs and other proteins related to MMP expression. CLSM was adopted for observing penetration of nanoemulsion of astaxanthin and showed high effective penetration rate compared to the nanoemulsion of astaxanthin prepared by conventional lecithin. In-vivo measurement of the nanoemulsions in hydration and elasticity were conducted to 11 Korean female adults for 28 days and showed better results.

Effect of Potassium Iodide on Erythrosine-Mediated Photodynamic Therapy on Streptococcus Mutans Biofilms (Streptococcus mutans 바이오필름에 대한 에리스로신 매개 광역동 치료 시 potassium iodide의 효과)

  • Yongsoon, Kim;Howon, Park;Juhyun, Lee;Haeni, Kim;Siyoung, Lee
    • Journal of the korean academy of Pediatric Dentistry
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    • v.49 no.3
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    • pp.321-328
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    • 2022
  • The aim of this in-vitro study is to evaluate the effect of potassium iodide (KI) on erythrosine-mediated photodynamic therapy (PDT) against Streptococcus mutans biofilms. S. mutans ATCC 25175 was cultured to form a biofilm on a hydroxyapatite disk. After diluting erythrosine to 20 μM and KI to 10, 50, and 100 mM, respectively, PDT was performed. The number of surviving bacteria was calculated as colony forming units (CFU)/mL and the statistical significance of the difference between groups was confirmed by Bonferroni post-hoc analysis. Cell viability was visually evaluated using confocal laser scanning microscopy (CLSM). As a result of the experiment, a significant decrease (p < 0.05) in CFU was observed in the experimental groups in which PDT was performed after applying KI regardless of the concentration of KI. In addition, a significant reduction (p < 0.05) in CFU was observed in the experimental group to which 100 mM KI was applied compared to 10 mM KI. The same results were confirmed when observing CLSM. KI significantly improved the efficacy of erythrosine-mediated PDT on S. mutans biofilms at all concentrations. This may compensate for the low sensitivity of PDT to biofilm-state bacteria strains, but it is necessary to establish an optimal clinical protocol through further research.

COMPARATIVE STUDY ON THE EARLY DETECTION OF ENAMEL LESIONS USING DIFOTI AND LASER FLUORESCENCE (Digital Imaging Fiber-Optic Trans-Illumination과 Laser Fluorescence를 이용한 법랑질 우식증의 조기 진단에 관한 비교 연구)

  • Maeng, Myoung-Ho;Kim, Seung-Oh;Kim, Jong-Soo
    • Journal of the korean academy of Pediatric Dentistry
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    • v.33 no.2
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    • pp.207-220
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    • 2006
  • The newly developed equipments for the early detection of carious lesion are LFD (laser fluorescence device), Ultrasonic diagnostic system, CLSM(confocal laser scanning microscopy), QLF(quantitative light-induced fluorescence) and DIFOTI (digital imaging fiber-optic trans-illumination) system. In this study, DIFOTI system and LFD were used for the detection of early enamel caries. Twenty five primary teeth extracted from twenty one children at around the dentitional exchanging period were selected as samples. The results obtained from DIFOTI imaging and LFD measurement were compared with those of CLSM and comprehensive evaluations were made for the diagnostic capacity of each device. In vitro test, 40 sample teeth with their buccal & lingual surface formed by a window of $2{\times}3mm$ in diameter were immersed in artificial demineralizing solution for the period of 4, 8, 12 and 16 days. The results obtained from the experimental groups (DIFOTI, LFD) were compared to control group (CLSM) and we have reached to the following conclusions. 1. The sensitivity and specificity of DIFOTI system operated in oral environment was 88.2% and 76.9% respectively. 2. The sensitivity and specificity of LFD measured in oral environment was 76.5% and 69.2% respectively. 3, Regression analysis on the light transparent rate of DIFOTI showed its decrease according to the length of primary enamel decalcification performed in vitro(r=-0.96, p<0.05). 4. No statistically significant difference between LFT measurement and the length of in vitro decalcification was found in regression analysis (p>0.05). 5. The correlation coefficient of DIFOTI image transparent rate and the lesion depth of CLMS was -0.6988 (p<0.05), whereas no statistically significant difference was found for LFD measurement.

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Morphology and Adhesion of Campylobacter jejuni to Chicken Skin Under Varying Conditions

  • Jang, Keum-Il;Kim, Min-Gon;Ha, Sang-Do;Kim, Keun-Sung;Lee, Kyu-Ho;Chung, Duck-Hwa;Kim, Cheorl-Ho;Kim, Kwang-Yup
    • Journal of Microbiology and Biotechnology
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    • v.17 no.2
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    • pp.202-206
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    • 2007
  • The adhesion of Campylobacter jejuni to chicken skin, along with the associated morphological changes under aerobic conditions at 4, 25, and $37^{\circ}C$ and microaerobic ($O_2\;5%,\;CO_2\;10%,\;N_2\;85%$) conditions, were investigated using confocal laser scanning microscopy (CLSM), flow cytometry, and plate counting. The morphological change of C. jejuni from a spiral shape to a coccoid form or VBNC form (viable but nonculturable form) progressed rapidly under aerobic conditions at 25, 37, and $4^{\circ}C$. As regards adhesion, the C. jejuni cells were mostly located in the crevices and feather follicles of the chicken skin, where the cells in the feather follicles floated freely in the entrapped water, even after the skin was rinsed quite thoroughly. CLSM also revealed the penetration of some spiral-shaped C. jejuni cells into the chicken skin. Even after changing their shape at various temperatures, coccoid-form C. jejuni cells were still found in the crevices and feather follicles of the chicken skin.

The remineralization effect of topical fluoride agents using confocal laser scanning microscope on artificial enamel caries aspects of convergence observation (공초점 레이저 주사 현미경을 이용한 우치 인공우식법랑질에서 불소도포 전·후의 재광화 효과에 대한 융합적 관찰)

  • Lee, Min-Sun;Kim, Hyo-Jin
    • Journal of the Korea Convergence Society
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    • v.8 no.6
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    • pp.139-145
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    • 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.

Antibacterial and Antibiofilm Effect of Cell-Free Supernatant of Lactobacillus brevis KCCM 202399 Isolated from Korean Fermented Food against Streptococcus mutans KCTC 5458

  • Kim, Jong Ha;Jang, Hye Ji;Lee, Na-Kyoung;Paik, Hyun-Dong
    • Journal of Microbiology and Biotechnology
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    • v.32 no.1
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    • pp.56-63
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    • 2022
  • This study aims to determine the antibiofilm effect of cell-free supernatant (CFS) of Lactobacillus brevis strains against Streptococcus mutans strains. To study the antibiofilm mechanism against S. mutans strains, antibacterial effects, cell surface properties (auto-aggregation and cell surface hydrophobicity), exopolysaccharide (EPS) production, and morphological changes were examined. The antibiofilm effect of L. brevis KCCM 202399 CFS as morphological changes were evaluated by scanning electron microscopy (SEM) and confocal laser scanning microscopy (CLSM), compared with the control treatment. Among the L. brevis strains, L. brevis KCCM 202399 showed the highest antibiofilm effect on S. mutans KCTC 5458. The antibacterial effect of L. brevis KCCM 202399 against S. mutans KCTC 5458 was investigated using the deferred method (16.00 mm). The minimum inhibitory concentration of L. brevis KCCM 202399 against S. mutans KCTC 5458 was 25.00%. Compared with the control treatment, L. brevis KCCM 202399 CFS inhibited the bacterial adhesion of S. mutans KCTC 5458 by decreasing auto-aggregation, cell surface hydrophobicity, and EPS production (45.91%, 40.51%, and 67.44%, respectively). L. brevis KCCM 202399 CFS inhibited and eradicated the S. mutans KCTC 5458 biofilm. Therefore, these results suggest that L. brevis KCCM 202399 CFS may be used to develop oral health in the probiotic industry.

Comparative lectin binding patterns of Cochlodinium polykrikoides Margalef

  • Rhodes, Lesley L.;Cho, Yong-Chul;Cho, Eun-Seob
    • Journal of the korean society of oceanography
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    • v.35 no.3
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    • pp.153-157
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    • 2000
  • Four different FITC-conjugated lectins were used to visually evaluate lectin binding activity by optical staining quality using confocal laser scanning microscopy (CLSM) of Cochzodinium polykrikoides in nature (wild type) and culture (cultured type). Cells from the field and cultures treated with ConA fluoresced only at the outer cell wall, and the abundance and distribution of the fluorescent signal were similar. Treatment with PWM and HPA did not elicit fluorescence at the cell surface, but the wild type exposed to HPA showed greater binding than did the cultured cells, possibly due to greater concentrations of glucosamine. The wild type cells treated with LBL lectin showed a strong green fluorescence on the cell surface, whereas cultured cells did not. Signal intensity and abundance were greater than for any other lectins tested in this study. These results suggest that wild type and cultured type are significantly different based on surface sugar production. In particular, the wild type cells apear richer in galactosamine-like moieties. Neither glucose nor mannose-like moieties were present in either wild types or cultured cells.

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Photoresponsive Behavior of Liquid-Crystalline Networks

  • Yu, Yanlei;Ikeda, Tomiki;Nakano, Makoto
    • 한국정보디스플레이학회:학술대회논문집
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    • 2002.08a
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    • pp.34-37
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    • 2002
  • Freestanding azobenzene-containing liquidcrystalline network (LCN) films. with macroscopic uniaxial molecular alignment were prepared by insitu photopolymerization. By polarizing microscopy, fiber-like structures aligned in one direction were observed. Furthermore, with a confocal laser scanning microscope (CLSM), it was confirmed that the fiber-like structures were formed even in the bulk of the LCNs. Upon UV light irradiation to cause trans-cis photoisomerization of the azobenzene molecules, the LCNfilms underwent a significant and anisotropic bending toward the irradiation direction of UV light. When the bent LCNfilms were exposed to Vis light, unbending of the LCN films immediately took place and the initial flat LCN films were restored. This bending and unbending behavior of the LCN films could be repeated just by changing the wavelength of the irradiation light. It was suggested that the bending was induced by an absorption gradient which produced a volume difference between the front surface area and the bulk of the network films.

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Anti-biofilm Activity of Xanthorrhizol Isolated from Curcuma xanthorrhiza Roxb. against Bacterial Biofilms Formed by Saliva and Artificial Multi-species Oral Strains

  • Yanti, Yanti;Rukayadi, Yaya;Lee, Kwan-Hyoung;Han, Sung-Hwa;Hwang, Jae-Kwan
    • Food Science and Biotechnology
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    • v.18 no.2
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    • pp.556-560
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    • 2009
  • Xanthorrhizol, a sesquiterpene isolated from Curcuma xanthorrhiza Roxb., was used to investigate its effect on reducing the saliva and multi-species oral biofilms consisting of Streptococcus mutans, Streptococcus sanguis, and Actinomyces viscosus by anti-biofilm and confocal laser scanning microscopy (CLSM) assays. Xanthorrhizol exhibited significant antibiofilm activity in the dose- and time-dependent manners. Exposure to 2 and $5{\mu}g/mL$ xanthorrhizol for 30 min remained <50% of saliva and multi-species biofilms formed for 24 hr. In addition, exposure to $10{\mu}g/mL$ xanthorrhizol for 30 min reduced 65 and 77% of 24 hr saliva and multi-species oral biofilms, respectively. CLSM results visually demonstrated that xanthorrhizol reduced bacterial viability in the saliva and multi-species oral biofilms. These results suggest that C. xanthorrhiza Roxb. containing xanthorrhizol with strong anti-biofilm activity can be employed as a plant source for oral care functional foods.

Modulation of Electroosmotic Flow through Skin: Effect of Poly(Amidoamine) Dendrimers

  • Kim, Hye Ji;Oh, Seaung Youl
    • Biomolecules & Therapeutics
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    • v.26 no.2
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    • pp.182-190
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    • 2018
  • The objective of this work is to evaluate the effect of polyamidoamine (PAMAM) dendrimers on electroosmotic flow (EOF) through skin. The effect of size and concentration of dendrimer was studied, using generation 1, 4 and 7 dendrimer (G1, G4 and G7, respectively). As a marker molecule for the direction and magnitude of EOF, a neutral molecule, acetoaminophen (AAP) was used. The visualization of dendrimer permeation into the current conducting pore (CCP) of skin was made using G4-fluorescein isothiocyanate (FITC) conjugate and confocal microscopy. Without dendrimer, anodal flux of AAP was much higher than cathodal or passive flux. When G1 dendrimer was added, anodal flux decreased, presumably due to the decrease in EOF by the association of G1 dendrimer with net negative charge in CCP. As the generation increased, larger decrease in anodal flux was observed, and the direction of EOF was reversed. Small amount of methanol used for the preparation of dendrimer solution also contributed to the decrease in anodal flux of AAP. Cross-sectional view perpendicular to the skin surface by confocal laser scanning microscope (CLSM) study showed that G4 dendrimer-FITC conjugate (G4-FITC) can penetrate into the viable epidermis and dermis under anodal current. The permeation route seemed to be localized on hair follicle region. These results suggest that PAMAM dendrimers can permeate into CCP and change the magnitude and direction of EOF. Overall, we obtained a better understanding on the mechanistic insights into the electroosmosis phenomena and its role on flux during iontophoresis.