• Title/Summary/Keyword: Specific migration

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Effects of Extracellular Stimulation of Different Niche Condition on the Transcriptional Regulation of Matrix Metalloproteinase Genes in the Mouse Embryonic Stem Cells

  • Yun, Jung Im;Kim, Min Seong;Lee, Seung Tae
    • Reproductive and Developmental Biology
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    • v.37 no.2
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    • pp.79-83
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    • 2013
  • Matrix metalloproteinases (MMPs) have been known to affect to cell migration, proliferation, morphogenesis and apoptosis by degrading the extracellular matrix. In the previous studies, undifferentiated mouse embryonic stem cells (ESCs) were successfully proliferated inside the extracellular matrix (ECM) analog-conjugated three-dimensional (3D) poly ethylene glycol (PEG)-based hydrogel. However, there is no report about MMP secretion in ESCs, which makes it difficult to understand and explain how ESCs enlarge space and proliferate inside 3D PEG-based hydrogel constructed by crosslinkers containing MMP-specific cleavage peptide sequence. Therefore, we investigated what types of MMPs are released from undifferentiated ESCs and how extracellular signals derived from various niche conditions affect MMP expression of ESCs at the transcriptional level. Results showed that undifferentiated ESCs expressed specifically MMP2 and MMP3 mRNAs. Transcriptional up-regulation of MMP2 was caused by the 3D scaffold, and activation of integrin inside the 3D scaffold upregulated MMP2 mRNAs synergistically. Moreover, mouse embryonic fibroblasts (MEFs) on 2D matrix and 3D scaffold induced upregulation of MMP3 mRNAs, and activation of integrins through conjugation of extracellular matrix (ECM) analogs with 3D scaffold upregulated MMP3 mRNAs synergistically. These results suggest that successful proliferation of ESCs inside the 3D PEG-based hydrogel may be caused by increase of MMP2 and MMP3 expression resulting from 3D scaffold itself as well as activation of integrins inside the 3D PEG-based scaffold.

An Electron Microscopic Radioautographic Study of the Synthesis and Migration of the Glycoproteins in the Osteoclast of the Mice Maxillary Alveolar Bone (생쥐 상악치조부에서의 파골세포의 당단백 합성 및 이동에 관한 전자현미경 자기방사법적 연구)

  • Kim, Myung-Kook
    • Applied Microscopy
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    • v.22 no.2
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    • pp.118-126
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    • 1992
  • The pathway and time course of fucose-containing glycoprotein synthesis and intracellular translocation in osteoclasts of the mice maxillary alveolar bone were investigated by electron microscopic radioautography. Male Balb-C mice weighing 17gm were anesthetized with Nembutal and injected via the external jugular vein with 2.5 mCi of $L-[6-^{3}H]-fucose$ (specific activity 16.8 mCi/mmol) in 0.1 ml of sterile saline solution. At 5, 10, 20, 35 minutes and 8 hours after administration of the $^{3}H-fucose$, animals were killed by intracardiac perfusion of 30ml of 2% glutaraldehyde in a modified Tyroid solution, pH 7.4. The maxillae were then removed and further fixed in Karnovsky fixative for an additional 3-4 hours. After rinsing in 0.1M cacodylate buffer for 10 minutes, the maxillae were demineralized for 2 weeks at $4^{\circ}C$ in ethylene diamine tetra acetate containing 2% glutaraldehyde. The first interdental areas were mesiodistally sectioned into slices of 1mm thickness and postfixed in osmium tetroxide. Tissues were then dehydrated and embedded in Poly Bed. To prepare electron microscopic radioautography, the dipping method of Kopriwa (1973) was employed. Thin sections were coated with a crystalline monolayer of ILford $L_4$ photographic emulsion. After exposure for 4 months at $4^{\circ}C$, the sections were developed Kodak Microdol-X and Phenidon (for compact grains), fixed in 30% sodium thiosulfate, stained with uranyl acetate and lead citrate and examined in the electron microscope (JEOL 1200 EX). At 5, 10 and 20 minutes after injection, $^{3}H-fucose$ was concentrated in Golgi cisternae of the osteoblasts. By 35 minutes the labels were observed over small vesicles in the suprannclear area of osteoclasts. At 8 hours, numerous silver grains were located on the ruffled border and cell membrane of osteoclasts. These results indicate that fucose molecules are added in the Golgi apparatus and small vesicles appear to be responsible for translocation of the glycoproteins to the marginal portion of osteoblasts. The glycoproteins are distributed on the osteoclast cell surface and especially over the ruffled border.

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Anti-tumor Immunity Induced by Tumor Cells Expressing a Membrane-bound Form of IL-2 and SDF-1

  • Choi, Jin-Wha;Lim, Ho-Yong;Chang, Mi-Ra;Cheon, Ji-Yeon;Kim, Young-Sang
    • Animal cells and systems
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    • v.12 no.4
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    • pp.193-201
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    • 2008
  • The eventual goal of tumor immunotherapy is to develop a vaccine inducing a specific anti-tumor immunity. Cytokine gene therapy is an effective way at least in animal models, but limited efficacy and various side effects obstruct clinical applications. In this study, we developed a tumor vaccine expressing a membrane-bound form of IL-2(mbIL-2) and SDF-1 in B16F10 melanoma cells. The tumor clones expressing mbIL-2 showed reduced tumorigenicity, and additional expression of SDF-1 to mbIL-2 expressing tumor cells caused more severe reduction in tumorigenicity. However, expression of the SDF-1 alone did not affect on the tumorigenicity, probably because of limited production of SDF-1 in the SDF-1 transfected clones. When the mice once rejected mbIL-2/SDF-1 expressing tumor clone were re-challenged with wild type B16F10 tumor cells, all of the mice survived. This result suggests that mbIL-2/SDF-1 tumor clone is effective in inducing systemic anti-tumor immunity against wild type B16 melanoma. Furthermore, culture supernatant of tumor clones expressing SDF-1 induced lymphocyte migration in vitro. These results, all together, suggest that expression of mbIL-2 and SDF-1 in tumor cells enhances anti-tumor immune responses through different roles; the secreted SDF-1 may function as a chemoattractant to recruit immune cells to tumor vaccine injection site, and the mbIL-2 on tumor cells may provide costimulatory signal for CTL activation in physical contacts.

OWL/Relational Mapping Rules to Use Relational Databases as OWL 2 Web Ontologies (관계형 데이터베이스를 OWL 2 웹 온톨로지로 사용하기 위한 OWL/관계형 매핑 규칙)

  • Choi, Ji-Woong;Kim, Myung-Ho
    • Journal of the Korea Society of Computer and Information
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    • v.16 no.7
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    • pp.35-47
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    • 2011
  • This paper proposes a set of rules to automatically generate OWL ontologies from relational databases. The purpose of the rules is to allow semantic access to existing RDB data without any database schema transformation and data migration process. In other words, the rules help a RDBMS play as a web ontology repository as well. However, the use of the mapping rules between RDB and OWL proposed by other studies for the objective causes troubles as follows. First, databases including the tables with a specific structure can't be translated into OWL. Second, the process for extracting an OWL individual unnecessarily lead to database join operations, or several SQL queries. On the other hand, our rules is designed to prevent these problems, can generate OWL classes and properties from database schemas and can generate OWL individuals from the database instances. In addition, an ontology generated by our rules is an OWL 2 DL ontology.

DCT-based Digital Dropout Detection using SVM (SVM을 이용한 DCT 기반의 디지털 드롭아웃 검출)

  • Song, Gihun;Ryu, Byungyong;Kim, Jaemyun;Ahn, Kiok;Chae, Oksam
    • Journal of the Institute of Electronics and Information Engineers
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    • v.51 no.7
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    • pp.190-200
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    • 2014
  • The video-based system of the broadcasters and the video-related institutions have shifted from analogical to digital in worldwide. This migration process can generate a defect, digital dropout, in the quality of the contents. Moreover, there are limited researches focused on these kind of defects and those related have limitations. For that reason, we are proposing a new method for feature extraction emphasizing in the peculiar block pattern of digital dropout based on discrete cosine transform (DCT). For classification of error block, we utilize support vector machine (SVM) which can manage feature vectors efficiently. Further, the proposed method overcome the limitation of the previous one using continuity of frame by frame. It is using only the information of a single frame and works better even in the presence of fast moving objects, without the necessity of specific model or parameter estimation. Therefore, this approach is capable of detecting digital dropout only with minimal complexity.

Gintonin stimulates autophagic flux in primary cortical astrocytes

  • Rahman, Md. Ataur;Hwang, Hongik;Nah, Seung-Yeol;Rhim, Hyewhon
    • Journal of Ginseng Research
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    • v.44 no.1
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    • pp.67-78
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    • 2020
  • Background: Gintonin (GT), a novel ginseng-derived exogenous ligand of lysophosphatidic acid (LPA) receptors, has been shown to induce cell proliferation and migration in the hippocampus, regulate calcium-dependent ion channels in the astrocytes, and reduce β-amyloid plaque in the brain. However, whether GT influences autophagy in cortical astrocytes is not yet investigated. Methods: We examined the effect of GT on autophagy in primary cortical astrocytes using immunoblot and immunocytochemistry assays. Suppression of specific proteins was performed via siRNA. LC3 puncta was determined using confocal microscopy. Results: GT strongly upregulated autophagy marker LC3 by a concentration- as well as time-dependent manner via G protein-coupled LPA receptors. GT-induced autophagy was further confirmed by the formation of LC3 puncta. Interestingly, on pretreatment with an mammalian target of rapamycin (mTOR) inhibitor, rapamycin, GT further enhanced LC3-II and LC3 puncta expression. However, GT-induced autophagy was significantly attenuated by inhibition of autophagy by 3-methyladenine and knockdown Beclin-1, Atg5, and Atg7 gene expression. Importantly, when pretreated with a lysosomotropic agent, E-64d/peps A or bafilomycin A1, GT significantly increased the levels of LC3-II along with the formation of LC3 puncta. In addition, GT treatment enhanced autophagic flux, which led to an increase in lysosome-associated membrane protein 1 and degradation of ubiquitinated p62/SQSTM1. Conclusion: GT induces autophagy via mTOR-mediated pathway and elevates autophagic flux. This study demonstrates that GT can be used as an autophagy-inducing agent in cortical astrocytes.

Ni/Si/Ni Ohmic contacts to n-type 4H-SiC (Ni/Si/Ni n형 4H-SiC의 오옴성 접합)

  • 이주헌;양성준;노일호;김창교;조남인;정경화;김은동;김남균
    • Proceedings of the Korean Institute of Electrical and Electronic Material Engineers Conference
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    • 2001.11a
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    • pp.197-200
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    • 2001
  • In this letter, we report on the investigation of Ni/Si/Ni Ohmic contacts to n-type 4H-SiC. Ohmic contacts have been formed by a vacuum annealing and N$_2$ gas ambient annealing method at 950$^{\circ}C$ for 10 min. The specific contact resistivity($\rho$$\sub$c/), sheet resistance(R$\sub$S/), contact resistance(R$\sub$S/), transfer length(LT) were calculated from resistance(R$\sub$T/) versus contact spacing(d) measurements obtained from 10 TLM(transmission line method) structures. The resulting average values of vacuum annealing sample were $\rho$$\sub$c/=3.8x10$\^$-5/ Ω$\textrm{cm}^2$ , R$\sub$c/=4.9Ω, R$\sub$T/=9.8Ω and L$\sub$T/=15.5$\mu\textrm{m}$, resulting average values of another sample were $\rho$$\sub$c/=2.29x10$\^$-4/ Ω$\textrm{cm}^2$ , R$\sub$c/=12.9Ω, R$\sub$T/=25.8Ω. The Physical properties of contacts were examined using X-Ray Diffraction and Auger analysis, there was a uniform intermixing of the Si and Ni, migration of Ni into the SiC.

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Antiangiogenic and Antitumor Activities of the Cryptic Fragments with Kringle Architecture

  • Joe, Young-Ae;Kim, Myung-Rae;Shim, Byoung-Shik;Oh, Dae-Shik;Hong, Sung-Hee;Hong, Yong-Kil
    • Biomolecules & Therapeutics
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    • v.11 no.4
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    • pp.205-213
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    • 2003
  • Various angiogenesis inhibitors target vascular endothelial cells and block tumor angiogenesis. Angiostatin is a specific endogenous angiogenesis inhibitor in clinical trials, which contains only the first four triple loop structures, known as kringle domains. Its generated by proteolytic cleavage of its parent molecule plasminogen, which itself does not exhibit antiangiogenic activity. Kringle domains from prothrombin, apolipoprotein, hepatocyte growth factor, urokinase and tissue-type plasminogen activator also elicit anti-angiogenic or antitumor activities in several model systems, albeit low amino acid sequence identity between angiostatin and each individual kringle. However, the differential effects of each kringle domain on endothelial cell proliferation, and migration observed in these kringle domains, suggest that the amino acid sequence of the primary structure is still important although kringle architecture is essential for anti-mlgiogenic activity. If it is further studied as to how amino acid sequence and kringle architecture contributes in anti-angiogenic activity, with studies on underlying mechanisms of anti-angiogenesis by kringle-based angiogenesis inhibitors, it will provide basis for the development of new potent anti-angiogenesis inhibitors and improvement of the efficacy of angiogenesis inhibitors.

Studies on the phosphotyrsine-proteins in the rat cerbellar PSD fraction (흰쥐 소뇌 연접후치밀질내 phosphotryrosine 함유 단백질에 대한 연구)

  • 전일수;함소희;고복현
    • Journal of Life Science
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    • v.7 no.3
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    • pp.198-204
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    • 1997
  • The signal transduction through tyrosine kinases play important roles in neuronal development and synaptic regulation. We carried out immunoblot analyses to study tyrosine=phosphorylated proteins in the rat cerebellar postsynaptic density (PSD), a protein-rich cytoskeletal specialization underlying beneath the postsynaptic membrane. The overall protein composition of cerebellar PSD fractions was similar to that of the forebrain’s and only a few bands were different in Coomassie stain. Immunoblot analyses with phosphtyrosine-specific antiboy (4G10) showed that there are many more tyrosine-phosphorylated proteins in the cerebellar PSD than in the forebrain PSD. Interestiingly, a major phosphotyrosine signals in cerebellar PSD fractions was associated with a 50 kD molecular size, named as PSD-50. Migration of PSD-50 coincided with that of $\alpha$CaMKII and remained in the pellet fraction after N-octylglucoside extraction. These results indicate that tyrosine phosphorylation is important in cerebellar synaptic regulation and that the PSD-50 may be same as $\alpha$CaMKIIor a new protein which is a major substrate of tyrosine kinase.

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Hybrid PtCo Alloy Nanocatalysts Encapsulated by Porous Carbon Layers for Oxygen Reduction Reactions (다공성 탄소층이 코팅된 하이브리드 표면 구조를 갖는 산소 환원 반응용 PtCo 합금 나노 촉매)

  • Jang, Jeonghee;Sharma, Monika;Sung, Hukwang;Kim, Sunpyo;Jung, Namgee
    • Korean Journal of Materials Research
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    • v.28 no.11
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    • pp.646-652
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    • 2018
  • During a long-term operation of polymer electrolyte membrane fuel cells(PEMFCs), the fuel cell performance may degrade due to severe agglomeration and dissolution of metal nanoparticles in the cathode. To enhance the electrochemical durability of metal catalysts and to prevent the particle agglomeration in PEMFC operation, this paper proposes a hybrid catalyst structure composed of PtCo alloy nanoparticles encapsulated by porous carbon layers. In the hybrid catalyst structure, the dissolution and migration of PtCo nanoparticles can be effectively prevented by protective carbon shells. In addition, $O_2$ can properly penetrate the porous carbon layers and react on the active Pt surface, which ensures high catalytic activity for the oxygen reduction reaction. Although the hybrid catalyst has a much smaller active surface area due to the carbon encapsulation compared to a commercial Pt catalyst without a carbon layer, it has a much higher specific activity and significantly improved durability than the Pt catalyst. Therefore, it is expected that the designed hybrid catalyst concept will provide an interesting strategy for development of high-performance fuel cell catalysts.