• Title/Summary/Keyword: confocal

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Analysis of Enhancement Effect and Attachment Ability of Beneficial Intestinal Microflora in Puffed Grain Foods Using Confocal Laser Scanning Microscopy (곡물 소재 팽화식품에서 장내 유익균의 증진 효과 분석 및 공초점 현미경을 이용한 부착능 평가)

  • Jeong, Myeong-Kyo;Oh, Do-Geon;Kwon, Oh-Sung;Jeong, Jun-Young;Lee, Ym-Shik;Kim, Kwang-Yup
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.46 no.9
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    • pp.1071-1080
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    • 2017
  • This study examined the adhesiveness of beneficial intestinal bacteria to whole-grains using confocal scanning laser microscopy (CLSM), to demonstrate the prebiotic effects of whole-grains, and to develop prebiotic puffed snacks with these whole-grains. CLSM has been used to observe the adhesiveness of Lactobacillus acidophilus, which belongs to beneficial intestinal bacteria, to whole-grain powders using optical sectioning techniques. The enhanced effects on the growth of beneficial intestinal bacteria with the hot water grain extract were verified using an indirect count method. Finally, a puffed snack was produced with the prebiotic effect and the quality was evaluated by checking the chromaticity and degree of hardness. As a result, L. acidophilus exhibited adhesive ability to whole-grain powders and growth of selected beneficial intestinal bacteria were improved significantly. The Hunter L value of the developed puffed snack increased when seasoning was added. The hardness of the puffed snack with seasoning was higher than that of the control. The results of a sensory evaluation showed that the puffed snack with seasoning was highly rated in the overall preference compared to the control.

Enhancement of Immune Activities of Peptides from Asterias amurensis Using a Nano-encapsulation Process (나노 입자 불가사리 펩타이드의 면역 활성 증진)

  • Jeong, Hyang-Suk;Oh, Sung-Ho;Kim, Seoung-Seop;Jeong, Myoung-Hoon;Choi, Woon-Yong;Seo, Yong-Chang;Choi, Geun-Pyo;Kim, Jin-Chul;Lee, Hyeon-Yong
    • Korean Journal of Food Science and Technology
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    • v.42 no.4
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    • pp.424-430
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    • 2010
  • Immuno-modulatory activities of peptides from Asterias amurensis were investigated using a nano-encapsulation process. The molecular weights of the peptides in the range of 5-7 kDa were separated using Sephadex G-75 gel filtration. Eighty-five percent of the nano-particles were in the 300 nm range using dynamic light scattering. The cytotoxicity of the A. amurensis nano-particles against CCD-986sk human dermal fibroblast cells was 11.64% after adding 1.0 mg/mL of the samples, which was lower than that from the control (13.28% collagen). The secretion of $NO^-$ from macrophages was estimated as $40\;{\mu}M$ after adding 1.0 mg/mL of gelatin nano-particles, which was higher than the others. Prostaglandin $E_2$ production from UV-induced human skin cells decreased greatly to 860 pg/mL after adding 1.0 mg/mL of the samples. Confocal microscopy revealed that nano-particles effectively penetrated the cells within 1 hour. From these results, we consider that nano-encapsulation of the peptides from A. amurensis can improve their biological functions.

IN VITRO STUDY OF DEMINERALIZATION INHIBITION EFFECT AND FLUORIDE UPTAKE INTO ADJACENT TEETH OF LIGHT-CURED FLUORIDE-RELEASING RESTORATIVES (수종의 불소방출 수복재의 탈회억제 효과 및 불소침투에 관한 연구)

  • Kim, Song-Yi;Choi, Sung-Chul;Kim, Kwang-Chul;Choi, Yeong-Chul;Park, Jae-Hong
    • Journal of the korean academy of Pediatric Dentistry
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    • v.37 no.3
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    • pp.288-297
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    • 2010
  • The purpose of this study was to evaluate the effect of light cured fluoride-releasing materials on the inhibition of demineralization. In addition, the pattern of fluoride uptake of adjacent tooth structure was analyzed with EPMA. Eighty intact premolars were restored with $Filtek^{TM}$ Z250(control group, composite), Fuji Filling $LC^{TM}$(RMGI), Dyract $AP^{(R)}$ (compomer) and Beautifil II(giomer). Restored teeth were stored in distilled water for 30 days. Then sixty teeth(n=15) were exposed to demineralizing solution(pH 4.3). Demineralized teeth were bisected and polished. The specimens were observed with confocal laser scanning microscope. The depth of outer lesion and the thickness of inhibition zone were measured. Remained twenty teeth(n=5) were bisected for fluoride uptake analysis. The fluoride analysis were taken at enamel-restoration interface and dentin-restoration interface by electron probe micro-analyzer. The results are as follows: 1. The depth of outer lesion of Fuji Filling $LC^{TM}$ Dyract AP, Beautifil II was shallower than that of $Filtek^{TM}$ Z250 at the margin of restoration(p<0.05). 2. The thickness of caries inhibition zone of Fuji Filling $LC^{TM}$, Dyract AP, Beautifil II was greater than that of $Filtek^{TM}$ Z250 at the margin of restoration(p<0.05). 3. Fuji Filling $LC^{TM}$, Dyract AP, Beautifil II groups showed the greater fluoride uptake into enamel and dentine around restoration than $Filtek^{TM}$ Z250 group. 4. In dentin the difference of fluoride concentration were greater than in enamel, and Dyract AP showed the greatest fluoride concentration in dentin.

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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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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    • v.41 no.2
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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.

Photodynamic Therapy with Photofrin Reduces Invasiveness of U87 Malignant Human Glioma Cells (교모세포종 세포주 U87에서 Photofrin을 사용한 광역학 치료가 종양 침습성에 미치는 영향)

  • Woo, Hye Kyung;Cho, Kyung-Keun;Rha, Hyung Kyun;Lee, Kyung Jin;Park, Sung Chan;Cho, Jung Ki;Park, Hea Kwan;Kang, Joon Ki;Choi, Chang Rak
    • Journal of Korean Neurosurgical Society
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    • v.30 no.sup2
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    • pp.189-196
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    • 2001
  • Objective : We tested the hypothesis that photodynamic therapy(PDT) with Photofrin inhibits tumor invasion of U87 human glioma cells using several in vitro assay to measure tumor invasiveness. The effects of PDT on cell growth, directional migration and cell invasion were investigated. Material and Method : Tumor cells were treated with Photofrin at various doses and at a fixed optical(632nm) dose of $100mJ/cm^2$. Cytotoxicity was tested using the MTT method. Invasion assays including the matrigelartificial basement membrane barrier migration and spheroid confrontation with confocal microscopic analysis were used to study the relationship between PDT and invasiveness. Result : U87 cells showed a dose dependent cytotoxic response to increasing Photofrin dose. Data from the matrigel artificial basement membrane assay indicate that PDT inhibits the U87 cell migration dose dependently. Low doses of subcytotoxic PDT treatment, such as 2.5ug/ml Photofrin dose, also appeared to significantly inhibit migration of U87 cells(p<0.05). In co-cultures between U87 cell spheroids and brain aggregates, progressive invasion with destruction of the brain aggregate occurs. The extent of tumor cell infiltration and proportion or intact brain aggregate remaining after 24h differs in Photofrin PDT treated versus Photofrin only control, with changes suggestive of a dose-response effect. Conclusion : our data indicate that PDT with Photofrin significantly inhibits the invasiveness of U87 cells, and this inhibition is dose dependent.

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Effect of the Flavonoid Luteolin for Dextran Sodium Sulfate-induced Colitis in NF-${\kappa}B^{EGFP}$ Transgenic Mice (Dextran Sodium Sulfate 유발 장염 모델에서 루테올린의 치료효과)

  • Jang, Byung-Ik
    • Journal of Yeungnam Medical Science
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    • v.23 no.1
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    • pp.26-35
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    • 2006
  • Background: Luteolin, a flavone found in various Chinese herbal medicines is known to possess anti-inflammatory properties through its ability to inhibit various proinflammatory signaling pathways including NF-${\kappa}B$ and p38 MAPK. In this study, we investigated the potential therapeutic effect of luteolin on dextran sodium sulfate (DSS)-induced colitis. Materials and Methods: We used a transgenic mouse model expressing the enhanced green fluorescent protein (EGFP) under the transcriptional control of NF-${\kappa}B$ $cis$-elements. C57BL/6 NF-${\kappa}B^{EGFP}$ mice received 2.5% DSS in their drinking water for six days in combination with daily luteolin administration (1mg/kg body weight, 0.1ml vol, intragastric) or vehicle. NF-${\kappa}B$ activity was assessed macroscopically with a Charge-Coupled Device (CCD) camera and microscopically by confocal analysis. Results: A significant increase in the Disease Activity Index (DAI), histological score (p<0.05), IL-12 p40 secretion in colonic stripe culture (p<0.05) and EGFP expression was observed in luteolin and/or DSS-treated mice compared to water-treated mice. Interestingly, a trend toward a worse colitis (DAI, IL-12p40) was observed in luteolin-treated mice compared to non-treated DSS-exposed mice. In addition, EGFP expression (NF-${\kappa}B$ activity) strongly increased in the luteolin-treated mice compared to control mice. Confocal microscopy showed that EGFP positive cells were primarily lamina propria immune cells. Conclusions: These results suggest that luteolin is not a therapeutic alternative for intestinal inflammatory disorders derived for primary defects in barrier function. Thus, therapeutic intervention targeting these signaling pathways should be viewed with caution.

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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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Fabrication of PLGA/Dextran Double-Layered Microspheres by Oil-in-Water Solvent Evaporation Method (O/W 용매 증발법을 이용한 PLGA와 덱스트란의 이중층 미립구 제조)

  • Ko Jong Tae;Lee Jae-Ho;Lee Chang-Rae;Shin Hyung Sik;Yuk Soon Hong;Kim Moon Suk;Khang Gilson;Rhee John M.;Lee Hai Bang
    • Polymer(Korea)
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    • v.29 no.6
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    • pp.543-548
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    • 2005
  • Double-layered spheres play an important role in controlling drug delivery for pharmaceutical application, because of the low initial burst compared with single-layered spheres and targetable delivery to specific organ. But it has drawback in loading drug and controlling size. In this study, we developed double-layered spheres using relatively simple oil-in-water (O/W) solvent evaporation method witw/without ultrasonication and investigated the size variation of the double-layered microspheres on the contents of poly(lactide- co-glycolide) (PLGA). Double - layered spheres were char-acterized by scanning elecron microscope (SEM), camscope, and confocal fluorescence laser microscope (CFLM). Double-layered spheres showed smooth surfaces and obvious difference between core and corona by SEM observation and camscope. We observed the fluorescent core in the double-walled spheres composed of FlTC-dextran and PLGA using CFLM. It was found that the core of the microsphere was dextran and the corona of the fabricate microsphere was PLGA. Also, the more PLGA concentration, the more the size of the fabricating double-layered sphere observed.

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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