• Title/Summary/Keyword: Transmission Electron Microscopy

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Establishment and Characterization of Multipotent Germ Line Stem Cells (MGSCs) from Neonatal Mouse Testis (신생 생쥐 고환에서 기인한 다분화능 생식줄기세포주의 확립 및 특성 분석)

  • Han, Sang-Chul;Song, Haeng-Seok;Jun, Jin-Hyun
    • Clinical and Experimental Reproductive Medicine
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    • v.35 no.1
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    • pp.39-48
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    • 2008
  • Objective: The aim of this study was to investigate whether multipotent germline stem cells (MGSCs) can be established from neonatal mouse testis. Methods: Various cells containing MGSCs were collected from neonatal testis of ICR mice and allocated to plates for in vitro culture. After 7 days in culture, the cells were passed to a fresh culture plate and continuously cultured. From the third or fourth passage, the presumed MGSCs were cultured and maintained on mitomycin C-inactivated STO feeder cells. The MGSCs were cultured in a condition where mouse embryonic stem cells (ESCs) are cultured. Characteristics of the MGSCs were evaluated by RT-PCR, immunocytochemistry, alkaline phosphatase activity, karyotyping, and transmission electron microscopy. Results: Two MGSCs lines were established from 9 pooled sets of neonatal testicular cells. MGSCs colonies were morphologically undistinguishable from ESCs colonies and both MGSC lines as well as ESCs expressed undifferentiated stem cell markers, such as Thy-1, Oct-4, Nanog, Sox2 and alkaline phosphatase. Fine structure of undifferentiated MGSCs were similar to those of ESCs and 60% of MGSCs (12/20) had normal karyotype at passage 10. They were able to form embryoid bodies (EBs) and MGSC-derived EBs expressed marker genes of three germ layers. Conclusion: We could establish the MGSCs from neonatal mouse testis and they were differentiated to multipotent lineages of three germ layers. Molecular characteristics of MGSCs were similar to those of ESCs. Our results suggest a possibility that multipotent stem cells derived from testis, the MGSCs, could replace the ESCs in biotechnology and regenerative medicine.

Gene Silencing of β-catenin by RNAi Inhibits Proliferation of Human Esophageal Cancer Cells by Inducing G0/G1 Cell Cycle Arrest

  • Wang, Jin-Sheng;Ji, Ai-Fang;Wan, Hong-Jun;Lu, Ya-Li;Yang, Jian-Zhou;Ma, Li-Li;Wang, Yong-Jin;Wei, Wu
    • Asian Pacific Journal of Cancer Prevention
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    • v.13 no.6
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    • pp.2527-2532
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    • 2012
  • Objectives: The aim of the present study was to explore mechanisms underlying the effects of down-regulating ${\beta}$-catenin expression on esophageal carcinoma (EC) cells. Methods: Cell cycle distribution and apoptosis were determined using flow cytometry and annexin V apoptosis assay, respectively. Transmission electron microscopy (TEM) was used to examine changes in ultrastructure, while expression of cyclin D1 protein and mRNA was detected by western blot and real-time PCR. Proliferating cell nuclear antigen (PCNA) and extracellular signal-regulated kinase (ERK) 1-2 were evaluated by Western blot analysis. PCNA labeling index (LI) was determined by immunocytochemistry. Results: Compared with pGen-3-con transfected and Eca-109 cells, the percentage of G0/G1-phase pGen-3-CTNNB1 transfected cells was obviously increased (P<0.05), with no significant difference among the three groups with regard to apoptosis (P>0.05). pGen-3-CTNNB1 transfected cells exhibited obvious decrease in cyclin D1 mRNA and protein expression (P<0.05) and the ultrastructure of Eca-109 cells underwent a significant change after being transfected with pGen-3-CTNNB1, suggesting that down-regulating ${\beta}$-catenin expression can promote the differentiation and maturation. The expression of PCNA and the ERKI/2 phosphorylation state were also down-regulated in pGen-3-CTNNB1 transfected cells (P<0.05). At the same time, the PCNA labeling index was decreased accordingly (P<0.05). Conclusion: Inhibition of EC Eca-109 cellproliferation by down-regulating ${\beta}$-catenin expression could improve cell ultrastructure by mediating blockade in G0/G1 through inhibiting cyclin D1, PCNA and the MAPK pathway (p-ERK1/2).

Stimulative Effects of Hominis Placental Pharmacopuncture Solution Combined with Zinc-oxide Nanoparticles on RAW 264.7 Cells - ZnO HPPS more easily stimulates RAW 264.7 cells -

  • Hong, Tae-Keun;Kim, Jee-Hye;Woo, Ju-Youn;Ha, Ki-Tae;Joo, Myung-Soo;Hahn, Yoon-Bong;Jeong, Han-Sol
    • Journal of Pharmacopuncture
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    • v.15 no.3
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    • pp.13-18
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    • 2012
  • Objectives: The purpose of this study is to examine whether Hominis Placental pharmacopuncture solution (HPPS) combined with zinc-oxide nanoparticles (ZnO NP) activates RAW 264.7 cells. Methods: We soaked ZnO nanoparticles in the Hominis Placenta pharmacopuncture solution, thereby making a combined form (ZnO NP HPPS). The effect of ZnO NP HPPS on the intracellular reactive oxygen species (ROS) production was measured by 2', 7'-dichlorofluorescin diacetate (DCFH-DA) assay. The effect of ZnO NP HPPS on NF-${\kappa}B$ was measured by using a luciferase assay. The effect of ZnO NP HPPS on the cytokine expression was assessed by semi-quantitative reverse transcriptase polymerase chain reaction (RT-PCR). The cellular uptake of ZnO NP HPPS was measured by using a flow cytometric analysis, and cellular structural alterations were analyzed by using transmission electron microscopy (TEM). Results: Neither the HPPS nor the ZnO NPs induced intracellular ROS production in RAW 264.7 cells. Neither of the materials activated NF-${\kappa}B$ or it's dependent genes, such as TNF-${\alpha}$, IL-1, and MCP-1. However, ZnO NP HPPS, the combined form of ZnO NPs and HPPS, did induce the intracellular ROS production, as well as prominently activating NF-${\kappa}B$ and it's dependent genes. Also, compared to ZnO NPs, it effectively increa-sed the uptake by RAW 264.7 cells. In addition, cellular structural alterations were observed in groups treated with ZnO NP HPPS. Conclusions: Neither ZnO NP nor HPPS activated RAW 264.7 cells, which is likely due to a low cellular uptake. The ZnO NP HPPS, however, significantly activated NF-${\kappa}B$ and up-regulated its dependent genes such as TNF-${\alpha}$, IL-1, and MCP-1. ZnO NP HPPS was also more easily taken into the RAW 264.7 cells than either ZnO NP or HPPS.

Effect of Na Salt on the Formation of MgO Obtained from Mg(OH)2 by Precipitation Method (침전법으로 제조된 Mg(OH)2의 잔류 Na염이 MgO 입자 형성에 미치는 영향)

  • Lee, Dong-Hyun;Ryu, Seung-Bom;Kim, Dae-Sung;Lim, Hyung-Mi;Lee, Seung-Ho
    • Korean Journal of Materials Research
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    • v.22 no.1
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    • pp.54-60
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    • 2012
  • The particle size of MgO was examined as a function of the Na content in $Mg(OH)_2$ powders and the calcination temperature. $Mg(OH)_2$ suspension was obtained by dropwise precipitation of $Mg(NO_3)_2{\cdot}6H_2O$ and NaOH solutions. The suspension was diluted by varying the dilution volume ratio of distilled water to $Mg(OH)_2$ suspension to change the Na salt concentration in the suspension. $Mg(OH)_2$ slurry was filtered and dried at $60^{\circ}C$ under vacuum, and then its $Mg(OH)_2$ powder was calcined to produce MgO with different amount of Na content at $500\sim900^{\circ}C$ under air. Investigation of the physical and chemical properties of the various MgO powders with dilution ratio and calcination temperature variation was done by X-ray diffraction, transmission electron microscopy, BET specific surface area and thermal gravimetric analysis. It was observed that MgO particle size could depend on the condition of calcination temperature and dilution ratio of the $Mg(OH)_2$ suspension. The particle size of the MgO depends on the Na content remaining in the $Mg(OH)_2$ powder, which powder was prepared by changing the dilution ratio of the $Mg(OH)_2$ suspension. This change increased as the calcination temperature increased and decreased as the dilution ratio increased. The growth of MgO particle size according to the increase of temperature was more effective when there was a relatively high content of Na. The increase of Na content lowered the temperature at which decomposition of $Mg(OH)_2$ to MgO took place, thereby promoting the crystal growth of MgO.

Formation of Passivation Layer and Its Effect on the Defect Generation during Trench Etching (트렌티 식각시 식각 방지막의 형성과 이들이 결함 생성에 미치는 영향)

  • Lee, Ju-Wook;Kim, Sang-Gi;Kim, Jong-Dae;Koo, Jin-Gon;Lee, Jeong-Yong;Nam, Kee-Soo
    • Korean Journal of Materials Research
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    • v.8 no.7
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    • pp.634-640
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    • 1998
  • A well- shaped trench was investigated in view of the defect distribution along trench sidewall and bottom using high resolution transmission electron microscopy. The trench was formed by HBr plasma and additive gases in magnetically enhanced reactive ion etching system. Adding $0_2$ and other additive gases into HBr plasma makes it possible to eliminate sidewall undercut and lower surface roughness by forming the passivation layer of lateral etching, resulted in the well filled trench with oxide and polysilicon by subsequent deposition. The passivation layer of lateral etching was mainly composed of $SiO_xF_y$ $SiO_xBr_y$ confirmed by chemical analysis. It also affects the generation and distribution of lattice defects. Most of etch induced defects were found in the edge region of the trench bottom within the depth of 10$\AA$. They are generally decreased with the thickness of residue layer and almost disappeared below the uni¬formly thick residue layer. While the formation of crystalline defects in silicon substrate mainly depends on the incident angle and energy of etch species, the region of surface defects on the thickness of residue layer formed during trench etching.

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Effect of antibacterial effects of myrrh, rhatany, chamomomilla against to oral microorganisms (몰약, 라타니아, 카모밀레 등의 구강 내 병원균에 대한 항균작용)

  • Baek, Han-Seung;Kang, Soo-Kyung;Auh, Q-Schick;Chun, Yang-Hyun;Hong, Jung-Pyo
    • Journal of Oral Medicine and Pain
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    • v.38 no.4
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    • pp.299-312
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    • 2013
  • Even though there exist a lot of study about antibacterial effects and reactions of extracted materials from plant, few study exist about oral pathogenic bacteria. Therefore we tried to recognize about the suppression effect to the periodontal pathogenic bacteria and halitosis, when add some kinds of plant extracted materials, myrrh, rhatany, chamomolilla to saliva. We used Crude drug : Myrrh tincture (100mg/ml), Ratanhia tincture (100mg/ml), Chamomile tincture(100mg/ml). We inspected about the cariogenic bateriae, S. mutans GS5 and S. sobrinus 6715, periodontal pathogenic bacteria, P. gingivalis 2561, P. intermedia ATCC 25611, Candida albicans ATCC 18804, and E. feacalis ATCC 4083, then the result follow. The plant extracted material, myrrh, rhatany, chamomomilla, which have convergence effect, bacteriocidal effect and anti-inflammation effect, show an antibacterial effect and reaction to the oral pathogenic bacteria. And with treating rhatany that have the most strong antibacterial effect, through transmission electron microscopy we could see a severe morphologic change of bacteria. This means with the plant extracted material, we can suppress the oral harmful bacteria and prevent periodontal diseases, caries, halitosis and oral inflammations. And within the future studies for the improvement of oral hygiene, our result might be a clinical evidence.

A Study on the Perpendicular Magnetic Anisotropy of Co-Pt Alloy Thin Films Deposited by DC Magnetron Sputtening (직류 마그네트론 스퍼터링으로 형성한 Co-Pt 합금박막의 수직자화기구에 대한 연구)

  • 박성언;김기범
    • Journal of the Korean Magnetics Society
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    • v.4 no.3
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    • pp.263-271
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    • 1994
  • We have produced $Co_{1-x}Pt_{x}(X\;=\;0.53\;and\;0.75)$ alloy films by DC magnetron sputtering at various substrate temperatures and sputtering pressures. Sputter-deposited Co-Pt alloy films showed a strong (111) texture, and the degree of (111) texture of the as-deposited film was found to depend on the substrate temperature and Ar pressure. However, we observed that the degree of (111) texture did not affect the magnetic properties. In addition, we have investigated the effect of heat-treatment on magnetic properties of these films. While the magnetic properties of the $Co_{0.25}Pt_{0.75}$ alloy films showed no noticeable changes, the coercivities and the squarenesses of the $Co_{0.47}Pt_{0.53}$ alloy films were drastically increased by annealing. Structural analysis using transmission electron microscopy(TEM) and x-ray diffractornetry(XRD) revealed that $CoPt(L1_{0})$ and $CoPt_3(L1_{2})$ ordered phases, respectively, were formed, each with a strong (111) texture. By comparing the magnetic properties between $CoPt(L1_{0})$ and $CoPt_3(L1_{2})$ ordered phases in relation to the atomic arrangements in a unit cell, we conclude that the magnetic anisotropy in the Co-Pt alloy system depends mainly on the atomic arrangements of Co and Pt.

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Fate and Bioaccumulation of Zinc Oxide Nanoparticles in a Microcosm (산화아연 나노물질의 미소생태계 내 거동 및 생물축적)

  • Kim, Eunjeong;Lee, Jae-woo;Jo, Eunhye;Sung, Hwa Kyung;Yoo, Sun Kyoung;Kim, Kyung-tae;Shin, Yu-jin;Kim, Ji-eun;Park, Sun-Young;Eom, Ig-chun;Kim, Pilje
    • Journal of Environmental Health Sciences
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    • v.43 no.3
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    • pp.194-201
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    • 2017
  • Objectives: Zinc oxide nanoparticles (ZnO NPs) are widely used in various commercial products, but they are exposed to the environment and can induce toxicity. In this study, we investigated the environmental fate and bioaccumulation of ZnO NPs in a microcosm. Methods: The microcosm was composed of water, soil (Lufa Soil 2.2) and organisms (Oryzias latipes, Neocaridina denticulata, Semisulcospira libertina). Point five and 5 mg/L of ZnO NPs were exposed in the microcosm for 14 days. Total Zn concentrations were measured using an Inductively Coupled Plasma Mass Spectrometer (ICP-MS) and intracellular NPs were observed using Transmission Electron Microscopy (TEM). Results: In the initial stages of exposure, the Zn concentrations in water increased in all exposure groups and then decreased, while the Zn concentration in soil increased after three hours for the 5 mg/L solution. Zn concentrations also showed increasing trends in N. denticulata and S. libertina at 0.5 and 5 mg/L, and in O. latipes at 5 mg/L. Accumulation of NPs was found in the livers of O. latipes and hepatopancreas of N. denticulata and S. libertina. Conclusions: In the early stages of exposure, ZnO NPs remained in the water, and then were transported to the soil and test species. Unlike other species, total Zn concentrations in N. denticulata and S. libertina increased for both 0.5 mg/L and 5 mg/L. Therefore, ZnO NPs were more easily accumulated in zoobenthos than in fish.

Study on Low-Temperature Solid Oxide Fuel Cells Using Y-Doped BaZrO3 (Y-doped BaZrO3을 이용한 저온형 박막 연료전지 연구)

  • Chang, Ik-Whang;Ji, Sang-Hoon;Paek, Jun-Yeol;Lee, Yoon-Ho;Park, Tae-Hyun;Cha, Suk-Won
    • Transactions of the Korean Society of Mechanical Engineers B
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    • v.36 no.9
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    • pp.931-935
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    • 2012
  • In this study, we fabricate and investigate low-temperature solid oxide fuel cells with a ceramic substrate/porous metal/ceramic/porous metal structure. To realize low-temperature operation in solid oxide fuel cells, the membrane should be fabricated to have a thickness of the order of a few hundreds nanometers to minimize IR loss. Yttrium-doped barium zirconate (BYZ), a proton conductor, was used as the electrolyte. We deposited a 350-nm-thick Pt (anode) layer on a porous substrate by sputter deposition. We also deposited a 1-${\mu}m$-thick BYZ layer on the Pt anode using pulsed laser deposition (PLD). Finally, we deposited a 200-nm-thick Pt (cathode) layer on the BYZ electrolyte by sputter deposition. The open circuit voltage (OCV) is 0.806 V, and the maximum power density is 11.9 mW/$cm^2$ at $350^{\circ}C$. Even though a fully dense electrolyte is deposited via PLD, a cross-sectional transmission electron microscopy (TEM) image reveals many voids and defects.

Identification of Daphne Mottle Virus Isolated from Daphne odora, a New Member of the Genus Potyvirus (서향에서 분리한 신종 포티바이러스(Daphne Mottle Virus)의 동정)

  • Park, Chung Youl;Park, Jungan;Lee, Boo-Ja;Bak, Sangmin;Lee, Hong-Kyu;Kim, Jeong-Sun;Yoon, Youngnam;Suh, Sang Jae;Lee, Su-Heon
    • Research in Plant Disease
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    • v.22 no.1
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    • pp.59-63
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    • 2016
  • A new poty-like virus was isolated from plants of winter daphne (Daphne odora) that showed virus-like symptoms on leaves, from four regions of Korea during 2014. Filamentous-shaped particles were observed by transmission electron microscopy of preparations extracted from symptomatic leaves and examined by the direct negative stain method. RT-PCR assay showed that three samples were positive for both Cucumber mosaic virus and potyvirus, and only one sample was positive for potyvirus only. A BLAST comparison to partial sequences from helper-component proteinase, cylindrical inclusion and coat protein genes detected the highest nucleotide identity of 76%, 72%, and 72% with Daphne mosaic virus, respectively, levels below the potyvirus species discrimination threshold. The new potyvirus was isolated using indicator plants (Chenopodium amaranticolor), in which local lesions were produced. In this study, we identified a novel potyvirus from winter daphne, which we have named Daphne mottle virus (DapMoV).