• Title/Summary/Keyword: Confocal Microscope

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Effects of Salviae Miltiorrhizae Radix on Blood-Brain Barrier Impairment of ICH-Induced Rats (단삼(丹蔘)이 뇌조직출혈 흰쥐의 혈액뇌관문 손상에 미치는 영향)

  • Park, Chang-Hoon;Kim, Youn-Sub
    • The Korea Journal of Herbology
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    • v.29 no.1
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    • pp.19-26
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    • 2014
  • Objectives : This study was performed in order to evaluate the effects of Salviae Miltiorrhizae Radix (SMR) water extract against the cerebral hemorrhage and the blood-brain barrier (BBB) impairment in the intracerebral hemorrhage (ICH). Method : ICH was induced by the stereotaxic intrastriatal injection of bacterial collagenase type IV in Sprague-Dawley rats. SMR was orally given three times every 20 hours during 3 days after the ICH induction. Hematoma volume, water content of brain tissue and volume of evans blue leakage were examined. Myeloperoxidase (MPO) positive neutrophils and tumor necrosis factor-${\alpha}$ (TNF-${\alpha}$) were observed with immunofluorescence labeling and confocal microscope. Results : SMR significantly reduced the hematoma volume of the ICH-induced rat brain. SMR significantly reduced the water content of brain tissue of the ICH-induced rat brain. SMR reduced the percentage of the evans blue leakage around the hematoma on the caudate putamen compared to the ICH group, especially on the cerebral cortex. SMR significantly reduced the volume of the evans blue leakage level in the peri-hematoma regions of the ICH-induced rat brain. SMR significantly reduced MPO positive neutrophils in the peri-hematoma regions of the ICH-induced rat brain. SMR reduced the TNF-${\alpha}$ expression in peri-hematoma regions of the ICH-induced rat brain. TNF-${\alpha}$ immuno-labeled cells were coincided with MPO immuno-labeled neutrophils in peri-hematoma regions of the ICH-induced rat brain. Conclusion : These results suggest that SMR plays a protective role against the blood-brain barrier impairment in the ICH through suppression of inflammation in the rat brain tissues.

Tribological Characteristics of DLC-Coated Part in Compressor (컴프레서 부품의 DLC코팅 적용에 따른 트라이볼로지적 특성 연구)

  • Yoon, Joo-Yong;Seo, Kuk-Jin;Han, Jae-Ho;Chun, Jihwan;Song, Jiyoung;Koh, Youngdeog;Nam, Jahyun;Kim, Seonkyo;Kim, Dae-Eun
    • Tribology and Lubricants
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    • v.38 no.1
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    • pp.8-14
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    • 2022
  • Amorphous carbon (a-C) has excellent wear resistance and, therefore is used as a coating to protect numerous mechanical components to prolong their lifetimes. Among the a-C coatings, diamond-like carbon (DLC) and DLC-containing silicon (Si-DLC) receive extensive attention owing to their enhanced wear resistance and low frictional characteristics. In this study, the friction and wear characteristics of DLC and Si-DLC coatings are analyzed. For comparative analysis, DLC-coated and Si-DLC-coated vanes are utilized with the counterpart of a roller for the friction tests. Since the lubricated mechanical components are generally vulnerable to wear when a lubricant film does not form properly, friction tests are conducted under boundary lubrication conditions to promote wear. A cylinder-on-cylinder type tribometer is used to perform the friction tests with various normal load conditions. After the friction test, a 3D laser confocal microscope is used for quantifying the wear volume to calculate the wear rate of each specimen. Consequently, the DLC-coated specimen shows a lower coefficient of friction (COF) and wear rate than the specimen without the coating, while the Si-DLC coating shows a higher COF than the bare specimen. The results of this study are expected to contribute to improving the efficiency and reliability of compressors.

Evaluation of intracellular uptake of cyclic RGD peptides in integrin αvβ3-expressing tumor cells

  • Soyoung Lee;Young-Hwa Kim;In Ho Song;Ji Young Choi;Hyewon Youn;Byung Chul Lee;Sang Eun Kim
    • Journal of Radiopharmaceuticals and Molecular Probes
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    • v.6 no.2
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    • pp.92-101
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    • 2020
  • The cyclic Arg-Gly-Asp (cRGD) peptide is well-known as a binding molecule to the integrin αvβ3 receptor which is highly expressed on activated endothelial cells and new blood vessels in tumors. Although numerous results have been reported by the usage of cRGD peptide-based ligands for cancer diagnosis and therapy, the distinct mechanisms, and functions of cRGD-integrin binding to cancer cells are still being investigated. In this study, we evaluated the internalization efficacy of different types of cRGD peptides (monomer, dimer and tetramer form) in integrin αvβ3 overexpressing cancer cells. Western blot and flow cytometric analysis showed U87MG expresses highly integrin αvβ3, whereas CT-26 does not show integrin αvβ3 expression. Cytotoxicity assay indicated that all cRGD peptides (0-200 µM) had at least 70-80% of viability in U87MG cells. Fluorescence images showed cRGD dimer peptides have the highest cellular internalization compare to cRGD monomer and cRGD tetramer peptides. Additionally, transmission electron microscope results clearly visualized the endocytic internalization of integrin αvβ3 receptors and correlated with confocal microscopic results. These results support the rationale for the use of cRGD dimer peptides for imaging, diagnosis, or therapy of integrin αvβ3-rich glioblastoma.

Characterization of Oleogels and Oleogel Emulsions Made with Sucrose Ester and Ceramide as Mixed Gelators (수크로스 에스터와 세라마이드를 혼합 겔레이터로 사용한 올레오겔과 올레오겔 에멀젼의 특성 분석)

  • Dayeon Lee;Byung Suk Jin
    • Applied Chemistry for Engineering
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    • v.34 no.5
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    • pp.501-506
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    • 2023
  • Oleogel and oleogel emulsions of sunflower oil were prepared using sucrose ester and ceramide as mixed gelators. The crystal structure of the gelator in the oleogels and oleogel emulsion formulations was observed with a polarized optical microscope, and the dispersion form of water was confirmed with confocal laser scanning microscopy. Through the DSC thermogram analysis, it was confirmed that the crystal structure of ceramide disappeared when sucrose ester and ceramide were mixed, and the crystallinity of the mixed gelator increased further when water was added to the formulation. Changes in rheological properties such as viscosity and viscoelasticity according to the ratio of sucrose ester, ceramide, and water in the formulation were examined. As the content of ceramide and water increased, the viscosity, storage modulus, and loss modulus all increased, and the stability of the formulation also tended to increase.

Exosomes from Tension Force-Applied Periodontal Ligament Cells Promote Mesenchymal Stem Cell Recruitment by Altering microRNA Profiles

  • Maolin Chang;Qianrou Chen;Beike Wang;Zhen Zhang;Guangli Han
    • International Journal of Stem Cells
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    • v.16 no.2
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    • pp.202-214
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    • 2023
  • Background and Objectives: To investigate the role of exosomes from periodontal ligament cells (PDLCs) in bone marrow mesenchymal stem cell (BMSC) migration. Methods and Results: Human PDLCs were applied cyclic tension stretching. Exosomes were extracted from cultured PDLCs by ultracentrifugation, then characterized for their size, morphology and protein markers by NTA, TEM and western blotting. The process that PKH26-labeled exosomes taken up by BMSCs was assessed by confocal microscope. BMSC migration was examined by Transwell assay. Exosomes derived from PDLCs were identified. Cyclic tension stretch application on PDLCs can enhance the migration ability of BMSCs through exosomes. The exosomal miRNA expression profiles of unstretched and stretched PDLCs were tested by miRNA microarray. Four miRNAs (miR-4633-5p, miR-30c-5p, miR-371a-3p and let-7b-3p) were upregulated and six (miR-4689, miR-8485, miR-4655-3p, miR-4672, miR-3180-5p and miR-4476) were downregulated in the exosomes after stretching. Sixteen hub proteins were found in the miRNA-mRNA network. Gene Ontology and KEGG pathway analyses demonstrated that the target genes of differentially expressed exosomal miRNAs closely related to the PI3K pathway and vesicle transmission. Conclusions: The exosomes derived from cyclic tension-stretched PDLCs can promote the migration of BMSCs. Alternation of microRNA profiles provides a basis for further research on the regulatory function of the exosomal miRNAs of PDLCs during orthodontic tooth movement.

Heat-Killed Enterococcus faecalis Prevents Adipogenesis and High Fat Diet-Induced Obesity by Inhibition of Lipid Accumulation through Inhibiting C/EBP-α and PPAR-γ in the Insulin Signaling Pathway

  • Jin-Ho Lee;Keun-Jung Woo;Min-Ah Kim;Joonpyo Hong;Jihee Kim;Sun-Hong Kim;Kwon-Il Han;Masahiro Iwasa;Tack-Joong Kim
    • Journal of Web Engineering
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    • v.14 no.6
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    • pp.1308-1322
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    • 2022
  • Increasing consumption of food with high caloric density and a sedentary lifestyle have influenced the increasing obesity prevalence worldwide. The recent pandemic has contributed to this problem. Obesity refers to a state in which lipid accumulates excessively in adipocytes and adipose tissues. Dried heat-killed Enterococcus faecalis (EF-2001) prevents allergic mechanisms, inflammation, and tumor progression. In the present study, we investigated the effects of EF-2001 on high fat diet (HFD)-induced obese rats. The degree of obesity in experimental rats was reduced after 6 weeks of oral administration of 3 mg/kg or 30 mg/kg dosages of EF-2001, indicating regulating effects in rats with HFD-induced obesity. We found that EF-2001 decreased the amounts of total cholesterol, triglyceride, and non-high density lipoprotein (HDL) in HFD-induced obese rats. The effects of EF-2001 on 3T3-L1 adipocytes stained with Oil red O stain are shown in reductions of lipid accumulation, respectively. In addition, we examined the relationships between EF-2001 treatment and mechanisms for the insulin signaling of adipogenesis in 3T3-L1 cells. EF-2001 induced down-regulation in phosphorylation of Erk, JNK, and Akt through the inhibition of insulin receptor phosphorylation. EF-2001 inhibits the expressions of C/EBP-α and PPAR-γ, a lipid metabolism-related transcription factor through confocal microscope observation and Western blot on 3T3-L1 adipocytes and HFD-induced obese rats. Based on our results, intake of EF-2001 significantly prevented HFD-induced obesity in rats through inhibition of C/EBP-α and PPAR-γ in the insulin signaling pathway on lipid accumulation.

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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Adhesion properties of Microorganisms onto surfaces of phosphorylcholine(PC)-modified copolymer for sensor applications (센서 응용을 위해 포스포릴 콜린으로 개질된 고분자 막 표면의 미생물 점착 특성)

  • Kim, Sun-Yong;Sohn, Ok-Jae;Chae, Kyu-Ho;Rhee, Jong-II
    • KSBB Journal
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    • v.23 no.3
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    • pp.226-230
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    • 2008
  • In this study we have studied adhesive properties of various microorganisms onto surfaces of phosphorylcholine-based copolymer for the application of optical biosensors. Three microorganisms, E.coli JM109, B.cereus 318, P.pastoris X-33 were cultivated in confocal cultivation dishes with glass surface, respectively. The glass surface was coated with copolymer containing 0% 5% and 10% MPC (2-methacryloxyethyl phosphorylcholine). After cultivation, culture medium was discarded and adhered microorganisms were dyed by gram staining method. Adhered microorganisms were analyzed using an optical microscope and scanning electronic microscope (SEM). A great number of microorganisms, $2-3{\times}10^3/mm^2$ were adhered on the surfaces of glass and copolymer membrane without MPC. But the antifouling effects of copolymer containing 5% and 10% phosphorylcholine were large, that microorganisms of less than $50-100/mm^2$ were attached on the copolymer membranes. Thus, the copolymer containing phosphorylcholine is very useful as an antifouling coating material for optical biosensor.

A study on the differentiation of MC3T3-E1 incubated on the layer-built silica/polycaprolactone non-woven fabric produced by electrospinning (전기방사법으로 제조된 실리카/폴리카프로락톤 적층형 부직포에 배양한 골아 세포의 중식, 분화에 관한 연구)

  • AN, Min-Kuk;Kim, Kyoung-Hwa;Kim, Tae-II;Lee, Yong-Moo;Rhee, Sang-Hoon;Ku, Young;Rhyu, In-Chul;Chung, Chong-Pyoung;Han, Soo-Boo;Seol, Yang-Jo
    • Journal of Periodontal and Implant Science
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    • v.37 no.1
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    • pp.115-124
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    • 2007
  • Silica is known as a promising osteoconductive material, and polycaprolactone is a bioactive and degradable material. The purpose of this study was to monitor the differentiation of MC3T3-E1 cells cultured on the layer-built silica/poly caprolactone non-woven fabric produced by electrospinning. Non-woven fabric (silica, polycaprolactone, PSP, SPS) was made by electrospinning and they were inserted in the 48 well cell culture plate. MC3T3-E1 cells were prepared by subculture. Cells were seeded to each well $1{\times}10^5$ concentration per well. Dulbecco's modified eagle medium with 10% FBS and 1% antibiotic-antimycotic solution was used. Confocal laser scanning microscope was taken 4 hours after incubation (95% air. 5% $CO_2$, $37^{\circ}C$). Cell proliferation was monitored by spectrophotometer on 1, 7, 14 days, and the morphology of the growing cells was observed by field emission scanning electron microscope. To monitor the differentiation of osteoblasts on the materials, MC3T3-E1 cells were incubated in 48 well culture plate after seeding with the density of $1{\times}10^5$ concentration. Then ELISA kit & EIA kit were used on to assess osteocalcin and osteopontin expression respectively. The other conditions were the same as above. MC3T3-E1 cells were proliferated well on all of the materials. There were no statistical differences among them. The osteopontin expression of silica, PSP, SPS was significantly higher than other groups on day 3 (p/0,05), but after that time, there were no statistically signigicant differences. The osteocalcin expression was significantly higher in silica and PSP than other groups on day 14. These findings show that PSP was as good as silica on the effect of osteoblast differentiation. The PSP non-woven fabric may have the possibility as bone graft materials.