• Title/Summary/Keyword: laser confocal system

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Interaction between the p75 neurotrophin receptor and a novel adaptor protein

  • Lee, Yun-Hee;Yu, Ji-Hee;Cho, Jung-Sun;Park, Han-Jeong;Lee, Seung-Pyo;Paik, Ki-Suk;Chang, Mi-Sook
    • International Journal of Oral Biology
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    • v.33 no.2
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    • pp.71-76
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    • 2008
  • The neurotrophin plays an important role in the development, differentiation and survival of the nervous system in vertebrates. It exerts its cellular effects through two different receptors, the Trk receptor tyrosine kinase neurotrophin receptor and the p75 neurotrophin receptor, a member of the tumor necrosis factor receptor superfamily. Trk and p75 neurotrophin receptors utilize specific target proteins to transmit signals into the cell. An ankyrin-rich membrane spanning protein (ARMS) was identified as a new p75 interacting protein and serves as a novel downstream target of p75 neurotrophin receptor. We sought to delineate the interaction between p75 and ARMS by deletion constructs of p75 and green fluorescent protein (GFP)-tagged ARMS. We examined the interaction between these two proteins after overexpressing them in HEK-293 cells. Using both Western blot analysis and immunocytochemistry followed by confocal laser scanning microscopy, we found out that the intracellular domain of the p75 neurotrophin receptor was important for the interaction with ARMS. The results from this study suggest that ARMS may play an important role for mediating the signals from p75 neurotrophin receptor into the cell.

GAP JUNCTION, A BIOMARKER FOR CANCER AND CHEMOPREVENTION: PREVENTIVE EFFECT OF EPICATECHIN AND GINSENOSIDE $Rb_$ ON THE INHIBITION OF GAP JUNCTIONAL INTERCULLULAR COMMUNICATION BY TPA AND $H_2O_2$

  • Kang, Kyung-Sun;Lee, Yong-Soom
    • Proceedings of the Korean Society of Toxicology Conference
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    • 2002.05b
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    • pp.59-72
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    • 2002
  • The anticarcinogenic effects of epicatechin(EC) and ginsenoside Rb2(Rb2), which are major components of green tea and Korea ginsen, respectively, were investigated using a model system of gap junctional intercellular communication (GJIC) in WB-F344 rat liver epithelial cells. 12-O-Tetradecanoylphorbol-13-accetate (TPA) and hydrogen preoxide, known as cancer promoters, inhibited GJIC in the epithelial cells as determined by the scrape loading/dye transfer assay, fluorescence redistribution assay after photobleaching, and immunofluorescent staining of connexin 43 using a laser confocal microscope. The inhibition of GJIC by TPA and H2O2 was prevented with treatment of Rb2 or Ec. The effect of EC on GJIC was stronger in TPA-treated cells than in H2O2-treated cells, while the effect of Rb2 was opoosite to that of EC. EC, at the concentration of 27.8$\mu$g/ml, prevented the TPA-induced GJIC inhibition by about 60%. Rb2, at the concentration of 277$\mu$g/ml, recovered the H2O2-induced GJIC inhibition by about 60%. These results suggest that Rb2 and EC may prevent human cancers by preventing the down-regulation of GJIC during the cancer promotion phase and that the anticancer effect of green tea and Korea ginseng may come from the major respective conponents, EC and Rb2.

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Recombinant Human Epidermal Growth Factor (rhEGF)-loaded Solid Lipid Nanoparticles: Fabrication and Their Skin Accumulation Properties for Topical rhEGF Delivery

  • Hwang, Hee-Jin;Han, Sunhui;Jeon, Sangok;Seo, Joeun;Oh, Dongho;Cho, Seong-Wan;Choi, Young Wook;Lee, Sangkil
    • Bulletin of the Korean Chemical Society
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    • v.35 no.8
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    • pp.2290-2294
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    • 2014
  • For the present study, rhEGF was encapsulated into solid lipid nanoparticles (SLNs). The SLNs were prepared by the $W_1/O/W_2$ double emulsification method combined with the high pressure homogenization method and the physical properties such as particle size, zeta-potential and encapsulation efficiency were measured. The overall particle morphology of SLNs was investigated using a transmission electron microscopy (TEM). The percutaneous skin permeation and accumulation property of rhEGF was evaluated using Franz diffusion cell system along with confocal laser scanning microscopy (CLSM). The mean particle size of rhEGF-loaded SLNs was $104.00{\pm}3.99nm$ and the zeta-potential value was in the range of -$36.99{\pm}0.54mV$, providing a good colloidal stability. The TEM image revealed a spherical shape of SLNs about 100 nm and the encapsulation efficiency was $18.47{\pm}0.22%$. The skin accumulation of rhEGF was enhanced by SLNs. CLSM image analysis provided that the rhEGF rat skin accumulation is facilitated by an entry of SLNs through the pores of skin.

Colonization and Population Changes of a Biocontrol Agent, Paenibacillus polymyxa E681, in Seeds and Roots

  • Park, Okhee;Kim, Jinwoo;Ryu, Choong-Min;Park, Chang-Seuk
    • The Plant Pathology Journal
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    • v.20 no.2
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    • pp.97-102
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    • 2004
  • Paenibacillus polymyxa E681, with its plant growth promotion and root colonization ability, has been proven to be a promising biocontrol agent of cucumber and barley. This study investigated the attributes related to the movement of bacteria from the seed to the radicle and to the whole root system. It also illustrated the existing form and population changes of the bacteria on seed and root using the scanning electron microscope and confocal laser scanning microscopy. The bacteria invaded and colonized the inside of the seed coat while the seeds were soaked in bacterial suspension. Almost the same number of bacteria on seed surface invaded the inside of the seed coat right after seed soaking. The population densities of E681 increased greatly inside as well as on the surface of the seed before the radicle emerged. The bacteria attached on the emerging radicle directly affected the initial population of newly emerg-ing root. The colonized cells on the root were arranged linearly toward the elongation of the root axis. In addition to colonizing the root surface, strain E681 was found inside the roots, where cells colonized the inter-cellular space between certain epidermal and cortical cells. When the cucumber seeds were soaked in bacterial suspension and sown in pot, the bacterial populations attached on both the surface and inside of the root were sustained up to harvesting time. This means that E681 successfully colonized the root of cucumber and sustained its population density up to harvesting time through seed treatment.

Physicochemical Characterization and Carcinoma Cell Interaction of Self-Organized Nanogels Prepared from Polysaccharide/Biotin Conjugates for Development of Anticancer Drug Carrier

  • Park Keun-Hong;Kang Dong-Min;Na Kun
    • Journal of Microbiology and Biotechnology
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    • v.16 no.9
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    • pp.1369-1376
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    • 2006
  • Self-organized nanogels were prepared from pullulan/biotin conjugates (PU/Bio) for the development of an effective anticancer drug delivery system. The degree of biotin substitution was 11, 19, and 24 biotin groups per 100 anhydroglucose units of pullulan. The physicochemical properties of the nanogels (PU/Bio1, 2 and 3) in aqueous media were characterized by dynamic light scattering, transmission electron microscopy, and fluorescence spectroscopy. The mean diameter of all the samples was less than 300 nm with a unimodal size distribution. The critical aggregation concentrations (CACs) of the nanoparticles in distilled water were $2.8{\times}10^{-2},\;1.6{\times}10^{-2}$, and $0.7{\times}10^{-2}mg/ml$ for the PU/Bio1, 2, and 3, respectively. The aggregation behavior of the nanogels indicated that biotin can perform as a hydrophobic moiety. To observe the specific interaction with a hepatic carcinoma cell line (HepG2), the conjugates were labeled with rhodamine B isothiocyanate (RITC) and their intensities measured using a fluorescence microplate reader. The HepG2 cells treated with the fluorescence-labeled PU/Bio nanoparticles were strongly luminated compared with the control (pullulan). Confocal laser microscopy also confirmed internalization of the PU/Bio nanogels into the cancer cells. Such results demonstrated that the biotin in the conjugate acted as both a hydrophobic moiety for self-assembly and a tumor-targeting moiety for specific interaction with tumor cells. Consequently, PU/Bio nanogels would appear to be a useful drug carrier for the treatment of liver cancer.

Push-out bond strength and dentinal tubule penetration of different root canal sealers used with coated core materials

  • Sungur, Derya Deniz;Purali, Nuhan;Cosgun, Erdal;Calt, Semra
    • Restorative Dentistry and Endodontics
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    • v.41 no.2
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    • pp.114-120
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    • 2016
  • Objectives: The aim of this study was to compare the push-out bond strength and dentinal tubule penetration of root canal sealers used with coated core materials and conventional gutta-percha. Materials and Methods: A total of 72 single-rooted human mandibular incisors were instrumented with NiTi rotary files with irrigation of 2.5% NaOCl. The smear layer was removed with 17% ethylenediaminetetraacetic acid (EDTA). Specimens were assigned into four groups according to the obturation system: Group 1, EndoRez (Ultradent Product Inc.); Group 2, Activ GP (Brasseler); Group 3, SmartSeal (DFRP Ltd. Villa Farm); Group 4, AH 26 (Dentsply de Trey)/gutta-percha (GP). For push-out bond strength measurement, two horizontal slices were obtained from each specimen (n = 20). To compare dentinal tubule penetration, remaining 32 roots assigned to 4 groups as above were obturated with 0.1% Rhodamine B labeled sealers. One horizontal slice was obtained from the middle third of each specimen (n = 8) and scanned under confocal laser scanning electron microscope. Tubule penetration area, depth, and percentage were measured. Kruskall-Wallis test was used for statistical analysis. Results: EndoRez showed significantly lower push-out bond strength than the others (p < 0.05). No significant difference was found amongst the groups in terms of percentage of sealer penetration. SmartSeal showed the least penetration than the others (p < 0.05). Conclusions: The bond strength and sealer penetration of resin-and glass ionomer-based sealers used with coated core was not superior to resin-based sealer used with conventional GP. Dentinal tubule penetration has limited effect on bond strength. The use of conventional GP with sealer seems to be sufficient in terms of push-out bond strength.

An experimental study of cutting efficiency of air-driven diamond burs on human tooth (수종 air-turbine 다이아몬드 버의 절삭 효과에 관한 실험적 연구)

  • Hong, Jin-Sun;Yeo, In-Sung;Kim, Sung-Hun;Lee, Jai-Bong;Han, Jung-Suk;Yang, Jae-Ho
    • The Journal of Korean Academy of Prosthodontics
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    • v.49 no.1
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    • pp.1-7
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    • 2011
  • Purpose: The purpose of this study was to investigate the cutting efficiency of coarse grit diamond burs with air-turbine handpiece on natural tooth. Materials and methods: Four groups of coarse grit diamond bur were selected: Komet (A), Shofu (B), Premier (C), and Mani (D). The extracted maxillary central incisors were used, and ten cuts were made on each specimen, using the rotary diamond burs. The surface of each bur was measured at the upper, middle, and bottom of the bur with confocal laser scanning microscope and imaged with SEM. The data were analyzed with one-way ANOVA and t-test at the significance level of 0.05. Results: The surface roughness was measured. At the A diamond bur, the Sa values were $52.93\;{\mu}m$, $48.32\;{\mu}m$, $46.79\;{\mu}m$, $45.06\;{\mu}m$, and $43.43\;{\mu}m$ for control, test 1, 2, 3, and 4 respectively. The Sa values were $50.68\;{\mu}m$, $45.62\;{\mu}m$, $44.41\;{\mu}m$, $44.10\;{\mu}m$, and $42.46\;{\mu}m$ for B diamond bur, $58.02\;{\mu}m$, $55.53\;{\mu}m$, $52.22\;{\mu}m$, $48.26\;{\mu}m$, and $45.36\;{\mu}m$ for C diamond bur, and $50.11\;{\mu}m$, $46.73\;{\mu}m$, $45.46\;{\mu}m$, $42.58\;{\mu}m$, and $41.80\;{\mu}m$ for D diamond bur. Surface roughness after each bur use showed significant changes, but no significant difference was found in surface roughness change between bur systems. Conclusions: Surface roughness in the same bur system showed significant differences after each tooth preparation. However no statistically significant differences were found in surface roughness between bur systems. The SEM images between control and test 4 showed the abraded particles.

Influence of finish line design on the marginal fit of nonprecious metal alloy coping fabricated by 3D printing, milling and casting using CAD-CAM (CAD-CAM을 이용한 3D printing, milling, casting 방법의 비귀금속 코핑의 지대치 변연 적합도 연구)

  • Seo-Rahng Kim;Myung-Joo Kim;Ji-Man Park;Seong-Kyun Kim;Seong-Joo Heo;Jai-Young Koak
    • The Journal of Korean Academy of Prosthodontics
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    • v.61 no.1
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    • pp.1-17
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    • 2023
  • Purpose. The purpose of this study was to examine the correlation between the finish line designs and the marginal adaptation of nonprecious metal alloy coping produced by different digital manufacturing methods. Materials and methods. Nonprecious metal alloy copings were made respectively from each master model with three different methods; SLS, milling and casting by computer aided design and computer aided manufacturing (CAD-CAM). Twelve copings were made by each method resulting in 72 copings in total. The measurement was conducted at 40 determined reference points along the circumferential margin with the confocal laser scanning microscope at magnification ×150. Results. Mean values of marginal gap of laser sintered copings were 11.8 ± 7.4 ㎛ for deep chamfer margin and 6.3 ± 3.5 ㎛ for rounded shoulder margin and the difference between them was statistically significant (P < .0001). Mean values of marginal gap of casted copings were 18.8 ± 20.2 ㎛ for deep chamfer margin and 33 ± 20.5 ㎛ for rounded shoulder margin and the difference between them was significant (P = .0004). Conclusion. Within the limitation of this study, the following conclusions were drawn. 1. The variation of finish line design influences the marginal adaptation of laser sintered metal coping and casted metal coping. 2. Laser sintered copings with rounded shoulder margin had better marginal fit than deep chamfer margin. 3. Casted copings with deep chamfer margin had better marginal fit than rounded shoulder margin. 4. According to the manufacturing method, SLS system showed the best marginal fit among three different methods. Casting and milling method followed that in order.

Evaluation of Alginate Microspheres Prepared by Emulsion and Spray Method for Oral Vaccine Delivery System (유화법과 분무법에 의해 제조된 경구백신용 알긴산 마이크로스피어의 평가)

  • Jiang, Ge;Jee, Ung-Kil;Maeng, Pil-Jae;Hwang, Sung-Joo
    • Journal of Pharmaceutical Investigation
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    • v.31 no.4
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    • pp.241-256
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    • 2001
  • Alginate microspheres, containing fluorescein isothiocyanate-bovine serum albumin (FITC-BSA) or green fluorescent protein (GFP) were prepared and used as a model drug to develop the oral vaccine delivery system. The alginate microspheres were coated with poly-L-lysine or chitosan. Two methods, w/o-emulsion and spray, were used to prepare alginate microspheres. To optimize preparation conditions, effects of several factors on the particle size and particle morphology of microsphere, and loading efficiency of model antigen were investigated. In both preparation methods, the particle size and the loading efficiency were enhanced when the concentration of sodium alginate increased. In the w/o-emulsion preparation method, as the concentration of Span 80 was increased from 0.5% to 2%, the particle size was decreased, but the loading efficiency was increased. The higher the emulsification speed was, the smaller the particle size and loading efficiency were. The concentration of calcium chloride did not show any effect on the particle size and loading efficiency. In the spray preparation method, the particle size was increased as the nozzle pressure $(from\;1\;kgf/m^2\;to\;3\;kgf/m^2)$ and spray rate was raised. Increasing calcium chloride concentration (<7%) decreased the particle size, in contrast to no effect of calcium chloride concentration on the w/o-emulsion preparation method. Alginate microspheres prepared by two methods were different in the particle size and loading efficiency, the particle size of microspheres prepared by the spray method was about $2-6\;{\mu}m$, larger than that prepared by the w/o emulsion method $(about\;2{\mu}m)$, and the loading efficiency was also higher with spray method. Furthermore, drying process for the microspheres prepared by the spray was simpler and easier, compared with the w/o emulsion preparation. Therefore, the spray method was chosen to prepare alginate microspheres for further experiments. Release pattern of FITC-BSA in alginate microspheres was evaluated in simulated intestinal fluid and PBS (phosphate buffered saline). Dissolution rate of FITC-BSA from alginate/chitosan microsphere was lower than that from alginate microsphere and alginate/poly-L-lysine microsphere. By confocal laser scanning microscope, it was revealed that alginate/FITC-poly-L-lysine microspheres were present in close apposition epithelium of the Peyer's patches of rabbits following inoculation into lumen of intestine, which proved that microspheres could be taken up by Peyer's patch. In conclusion, it is suggested that alginate microsphere prepared by spray method, showing a particle size of & $10\;{\mu}m$ and a high loading efficiency, can be used as a model drug for the development of oral vaccine delivery system.

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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.