• Title/Summary/Keyword: Fluorescence Microscope

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Cytoprotective effect of rhamnetin on miconazole-induced H9c2 cell damage

  • Lee, Kang Pa;Kim, Jai-Eun;Park, Won-Hwan
    • Nutrition Research and Practice
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    • v.9 no.6
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    • pp.586-591
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    • 2015
  • BACKGROUND/OBJECTIVES: Reactive oxygen species (ROS) formation is closely related to miconazole-induced heart dysfunction. Although rhamnetin has antioxidant effects, it remained unknown whether it can protect against miconazole-induced cardiomyocyte apoptosis. Thus, we investigated the effects of rhamnetin on miconazole-stimulated H9c2 cell apoptosis. MATERIALS/METHODS: Cell morphology was observed by inverted microscope and cell viability was determined using a WelCount$^{TM}$ cell proliferation assay kit. Miconazole-induced ROS production was evaluated by fluorescence-activated cell sorting with 6-carboxy-2',7'-dichlorofluoroscein diacetate ($H_2DCF$-DA) stain. Immunoblot analysis was used to determine apurinic/apyrimidinic endonuclease 1 (APE/Ref-1) and cleaved cysteine-aspartic protease (caspase) 3 expression. NADPH oxidase levels were measured using real-time polymerase chain reaction. RESULTS: Miconazole (3 and $10{\mu}M$) induced abnormal morphological changes and cell death in H9c2 cells. Rhamnetin enhanced the viability of miconazole ($3{\mu}M$)-treated cells in a dose-dependent manner. Rhamnetin (1 and $3{\mu}M$) treatment downregulated cleaved caspase 3 and upregulated APE/Ref-1 expression in miconazole-stimulated cells. Additionally, rhamnetin significantly reduced ROS generation. CONCLUSIONS: Our data suggest that rhamnetin may have cytoprotective effects in miconazole-stimulated H9c2 cardiomyocytes via ROS inhibition. This effect most likely occurs through the upregulation of APE/Ref-1 and attenuation of hydrogen peroxide levels.

Clonazepam Release from Core-shell Type Nanoparticles In Vitro

  • Kim, Hyun-Jung;Jeong, Young-Il;Kim, Sung-Ho;Lee, Young-Moo;Cho, Chong-Su
    • Archives of Pharmacal Research
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    • v.20 no.4
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    • pp.324-329
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    • 1997
  • AB-type amphiphilic copolymers (abbreviated as LE) composed of poly (L-leucine) (PLL) as the A component and poly (ethylene oxide) (PEO) as the B component were synthesized by the ring-opening polymerization of L-leucine N-carboxy-anhydride initiated by methoxy polyoxyethylene amine $(Me-PEO-NH_2)$ and characterized. Core-shell type nanoparticles were prepared by the diafiltration method. Particle size distribution obtained by dynamic light scattering was dependent on PLL composition and the size for LE-1, LE-2 and LE-3 was $369.6{\pm}267$, $523.4{\pm}410$ and $561.2{\pm}364 nm$, respectively. Shapes of the nanoparticies observed by transmission electron microscope (TEM) were almostly spherical. The critical micelle concentration (CMC) of the nanoparticles determined by a fluorescence probe technique was dependent on the composition of hydrophobic PLL, and the CMC for LE-1, LE-2 and LE-3 was $2.0{\times}10^{-6},1.7{\times}10^{-6}$ and $1.5{\times}10^{-6}(mol/l) $, respectively. Clonazepam release from core-shell type nanoparticles in vitro was dependent on PLL composition and drug loading content.

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ADHESIVENESS EVALUATION OF ACTIVATED PLATELET USING Arg-Gly-Asp-Phe(RGDF)-IMMOBILIZED SURFACE

  • Kim, J.H.;Kim, H.J.;Kim, J.;Ryu, G.H.;Min, B.G.;Choe, T.B.
    • Proceedings of the KOSOMBE Conference
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    • v.1997 no.11
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    • pp.333-336
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    • 1997
  • The adhesion of activated and normal platelets to fibrinogen requires the receptor binding site of GPIIb/IIIa. These recognition sites exists in the A ${\alpha}$ chain(RGDS at 572-575 and RGDF at 95-98) and the carboxy-terminal of ${\gamma}$ chain (HHLGGAKQAGDV at 400-411) of fibrinogen. In this study, we developed RGDF-immobilized surface to detect the unctional state of platelet. RGDF-immobilized surface was prepared on the glass using photolithographic technology. Platelet adhesion to RGDF-immobilized surface was observed by staining platelets with mepacrine using a fluorescence microscope using mepacrine. Using the RGDF peptide of fragment E, we observed that the platelets pretreated with PGE1 interacted incompletely with RGDF-immobilized surface, whereas ADP activated platelets interacted with the surface extensively. These results show that the distinct selectivity of RGDF-immobilized micro-patterned surface can be used to detect the unctional state of platelets.

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Quantitative Visualization of Oxygen Transfer in Micro-channel using Micro-LIF Technique (마이크로 레이저 형광 여기법을 이용한 미세채널 내부에서의 산소 확산에 대한 정량적 가시화)

  • Chen, Juan;Kim, Hyun-Dong;Kim, Kyung-Chun
    • Journal of the Korean Society of Visualization
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    • v.10 no.1
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    • pp.34-39
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    • 2012
  • In the present study, oxygen transfer process across gas-liquid interface in a Y-shape micro-channel is quantitatively visualized using the micro laser induced fluorescence (${\mu}$-LIF) technique. Diffusion coefficient of Oxygen ($D_L$) is estimated based on the experimental results and compared to its theoretical value. Tris ruthenium (II) chloride hexahydrate was used as the oxygen quenchable fluorescent dye. A light-emitting diode (LED) with wavelength of 450 nm was used as the light source and phosphorescence images of fluorescent dye were captured by a CMOS high speed camera installed on the microscope system. Water having dissolved oxygen (DO) value of 0% and pure oxygen gas were injected into the Y-shaped microchannel by using a double loading syringe pump. In-situ pixel-by-pixel calibration was carried out to obtain Stern-Volmer plots over whole flow field. Instantaneous DO concentration fields were successfully mapped according to Stern-Volmer plots and DL was calculated as $2.0675{\times}10^{-9}\;m^2/s$.

A Study on the Removel of Metallic Impurities on Silicon Surface and Mechanism using Remote Hydrogen Plasma (리모트 수소 플라즈마를 이용한 Si 표면의 금속오명 제거)

  • Park, Myeong-Gu;An, Tae-Hang;Lee, Jong-Mu;Jeon, Hyeong-Tak;Ryu, Geun-Geol
    • Korean Journal of Materials Research
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    • v.6 no.7
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    • pp.661-670
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    • 1996
  • 리모트 수소 플라즈마를 이용하여 실리콘 웨이퍼 표면 위에 있는 금속불순물의 제거 및 제거기구에 관하여 조사하였다. 실리콘의 표면과 내부분석을 위하여 TXRF(total reflection x-ray fluorescence)와 SPV(surface photovoltage), AFM(atomic force microscope)을 사용하였다. TXRF 분석결과 리모트 수소 플라즈마가 금속오염물질 제거에 상당한 효과가 있는 것으로 나타났다. TXRF분석결과 리모트 수소 플라즈마가 금속오염물질 제거에 상당한 효과가 있는 것으로 나타났다. 리모트 수소플라트마 처리 후 금속오염은 금속원소의 종류에 따라 1010atoms/$\textrm{cm}^2$-1011atoms/$\textrm{cm}^2$수준이었다. SPV분석결과를 보면 수소 플라즈마 처리에 의해 minority carrier 수명이 전반적으로 증가하였다. AFM 분석을 통하여 수소 플라즈마 처리가 표면 손상을 일으키지 않으며 표면의 거칠기에 나쁜 영향을 미치지 않음을 알 수 있었다. 또한 본 실험에서 나타난 결과들을 종합해 볼 때 금속오염물의 제거기구는 자연산화막 혹은 수소로 passivate된 실리콘 웨이퍼 표면을 수소 플라즈마에서 발생된 수소원자가 실리콘표면을 약하게 에칭할 때 떨어져 나가는 'lift-off'가 유력한 것으로 판단된다.

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Rapid Topological Patterning of Poly(dimethylsiloxane) Microstructure (Poly(dimethylsiloxane) 미세 구조물의 신속한 기하학적 패터닝)

  • Kim, Bo-Yeol;Song, Hwan-Moon;Son, Young-A;Lee, Chang-Soo
    • Textile Coloration and Finishing
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    • v.20 no.1
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    • pp.8-15
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    • 2008
  • We presented the modified decal-transfer lithography (DTL) and light stamping lithography (LSL) as new powerful methods to generate patterns of poly(dimethylsiloxane) (PDMS) on the substrate. The microstructures of PDMS fabricated by covalent binding between PDMS and substrate had played as barrier to locally control wettability. The transfer mechanism of PDMS is cohesive mechanical failure (CMF) in DTL method. In the LSL method, the features of patterned PDMS are physically torn and transferred onto a substrate via UV-induced surface reaction that results in bonding between PDMS and substrate. Additionally we have exploited to generate the patterning of rhodamine B and quantum dots (QDs), which was accomplished by hydrophobic interaction between dyes and PDMS micropatterns. The topological analysis of micropatterning of PDMS were performed by atomic force microscopy (AFM), and the patterning of rhodamine B and quantum dots was clearly shown by optical and fluorescence microscope. Furthermore, it could be applied to surface guided flow patterns in microfluidic device because of control of surface wettability. The advantages of these methods are simple process, rapid transfer of PDMS, modulation of surface wettability, and control of various pattern size and shape. It may be applied to the fabrication of chemical sensor, display units, and microfluidic devices.

Generation of Reactive Oxygen Species in Bovine Somatic Cell Nuclear Transfer Embryos during Micromanipulation Procedures

  • Hwang, In-Sun;Bae, Hyo-Kyung;Park, Choon-Keun;Yang, Boo-Keun;Cheong, Hee-Tae
    • Reproductive and Developmental Biology
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    • v.36 no.1
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    • pp.49-53
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    • 2012
  • The present study was conducted to examine the generation of reactive oxygen species (ROS) during micromanipulation procedures in bovine somatic cell nuclear transfer (SCNT) embryos. Bovine enucleated oocytes were electrofused with donor cells, activated by a combination of Ca-ionophore and 6-dimethylaminopurine culture. Oocytes and embryos were stained in dichlorodihydrofluorescein diacetate or 3'-(p-hydroxyphenyl) fluorescein dye and the $H_2O_2$ or $^.OH$ radical levels were measured. $In$ $vitro$ fertilization (IVF) was performed for controls. The samples were examined with a fluorescent microscope, and fluorescence intensity was analyzed in each oocyte and embryo. The $H_2O_2$ and $^.OH$ radical levels of reconstituted oocytes were increased during manipulation (37.2~49.7 and 51.0~55.2 pixels, respectively) as compared to those of mature oocytes ($p$<0.05). During early $In$ $vitro$ culture, the ROS levels of SCNT embryos were significantly higher than those of IVF embryos ($p$<0.05). These results suggest that the cellular stress during micromanipulation procedures can generate the ROS in bovine SCNT embryos.

The Localization of Cytokeratin 19 and Vimentin in Sprague Dawley Albino Rat Skin Tissue

  • Kim, Tae Keun;Kim, Yong Joo;Min, Byoung Hoon;Kim, Soo Jin
    • Applied Microscopy
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    • v.44 no.1
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    • pp.15-20
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    • 2014
  • Cytokeratin 19 (CK19) expressed in epidermis of skin, bulge region of hair follicle, outermost layer of outer root sheath and proximal and distal to bulge. Vimentin is a fibrous protein that localized in cytoplasm of fibroblast and forms cytoskeleton to maintain shape of cell and nucleus. In this study, CK19 and vimentin in skin were confirmed with light, fluorescence and transmission electron microscope. As a result, CK19 was localized epidermis, hair follicles, outer root sheath and nucleus of Merkel's cell. However, vimentin was localized some epidermis, dermis, hypodermis and nucleus of Merkel's cell. The role of CK19 is self-renewal and homeostasis in skin. Also, hair follicle regeneration and hair growth is known to be related. It is supposed that required of structural proteins that make up cytoskeleton is increased. Thereby, expression of CK19 is increased. It is considered that vimentin localized in order to stabilize structure of cell and cytoskeleton of fibroblasts. Also, CK19 and vimentin present in nuclei of Merkel's cell, and to act as a fibrous protein that make up end of a nerve fiber present in Merkel's cell and paracrine function of Merkel's cell.

Drug Release from Thermo-Responsive Self-assembled Polymeric Micelles Composed of Cholic Acid and Poly(N-isopropylacrylamide)

  • Kim, In-Sook;Jeong, Young-Il;Lee, Yun-Ho;Kim, Sung-Ho
    • Archives of Pharmacal Research
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    • v.23 no.4
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    • pp.367-373
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    • 2000
  • Cholic acid, conjugated with amine-terminated poly(W-isopropylacrylamide) (abbreviated as CA/ATPNIPAAm), was synthesized by a N, N'-dicyclohexyl carbodiimide (DCC)-mediated coupling reaction. Self-assembled CA/ATPNIPAAm micelles were prepared by a diafiltration method in aqueous media. The CA/ATPNIPAAm micelles exhibited a lower critical solution temperature (LCST) at $31.5^{\circ}C$. Micelle sizes measured by photon correlation spectroscopy (PCS) were approximately 31.6 $\times$$\times$ 5.8 nm. The CA/ATPNIPAAm micelles were spherical and their thermal size transition was observed by transmission electron microscope (TEM). A fluorescence probe technique was used for determining the micelle formation behavior of CA/ATPNIPAAm in aqueous solutions using Pyrene as a hydrophobic Probe. The critical micelle concentration (CMC) was evaluated as $8.9{\times}0^{-2}$ g/L. A drug release study was performed using indomethacin (IN) as a hydrophobic model drug. The release kinetics of IN from the CA/ATPNIPAAm micelles revealed a thermo-sensitivity by the unique character of poly(N-isopropylacrylamide) i.e. the release rate was higher at $25^{\circ}C$ than at $37^{\circ}C$.

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Studies on the Multiplication and Induction of Hybrid Plant in Cremastra appendiculata Use the the Embryo and Tissue Culture (배배양 및 조직배양을 이용한 약난초 (Cremastra appendiculata) 의 증식과 잡종식물의 유도에 관한 연구)

  • 이정석;황백김영준
    • KSBB Journal
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    • v.5 no.1
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    • pp.43-47
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    • 1990
  • It was determined optimal Culture conditions and suitable growth regulators for seed germination, growth of callus, and protoplasts derived from cultured and mesophyll cells in Gremastra appendiculata. Induction of fusion between protoplasts of cultured and mesophyll cells was examined. The best conditions of seed germination and growth of callus were achieved on Hyponex medium contained plant growth regulators(2mg/l 2, 4-D, lmg/l Kinetin). Viability and regeneration of cell wall in protoplasts was determined with fluorescence microscope. Also, fused protoplasts were achieved by using PEG solution between protoplasts of cultured and mesophyll cells.

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