• Title/Summary/Keyword: cell junctions

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Substrate roughness induces the development of defective E-cadherin junctions in human gingival keratinocytes

  • Jin, Chengbiao;Lee, Gayoung;Oh, Changseok;Kim, Hyun Jung;Kim, Hyun-Man
    • Journal of Periodontal and Implant Science
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    • v.47 no.2
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    • pp.116-131
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    • 2017
  • Purpose: The entry of bacteria or harmful substances through the epithelial seal of human gingival keratinocytes (HGKs) in the junctional epithelium (JE) is blocked by specialized intercellular junctions such as E-cadherin junctions (ECJs). However, the influence of roughened substrates, which may occur due to apical migration of the JE, root planing, or peri-implantitis, on the development of the ECJs of HGKs remains largely unknown. Methods: HGKs were cultured on substrates with varying levels of roughness, which were prepared by rubbing hydrophobic polystyrene dishes with silicon carbide papers. The activity of c-Jun N-terminal kinase (JNK) was inhibited with SP600125 or by transfection with JNK short hairpin RNA. The development of intercellular junctions was analyzed using scanning electron microscopy or confocal laser scanning microscopy after immunohistochemical staining of the cells for E-cadherin. The expression level of phospho-JNK was assessed by immunoblotting. Results: HGKs developed tight intercellular junctions devoid of wide intercellular gaps on smooth substrates and on rough substrates with low-nanometer dimensions (average roughness $[Ra]=121.3{\pm}13.4nm$), although the ECJs of HGKs on rough substrates with low-nanometer dimensions developed later than those of HGKs on smooth substrates. In contrast, HGKs developed short intercellular junctions with wide intercellular gaps on rough substrates with mid- or high-nanometer dimensions ($Ra=505.3{\pm}115.3nm$, $867.0{\pm}168.6nm$). Notably, the stability of the ECJs was low on the rough substrates, as demonstrated by the rapid destruction of the cell junction following calcium depletion. Inhibition of JNK activity promoted ECJ development in HGKs. JNK was closely associated with cortical actin in the regulation of ECJs in HGKs. Conclusions: These results indicate that on rough substrates with nanometer dimensions, the ECJs of HGKs develop slowly or defectively, and that this effect can be reversed by inhibiting JNK.

Computer Simulation of Switching Characteristics and Magnetization Flop in Magnetic Tunnel Junctions Exchange Biased by Synthetic Antiferromagnets

  • Lim, S.H.;Uhm, Y.R.
    • Journal of Magnetics
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    • v.6 no.4
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    • pp.132-141
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    • 2001
  • The switching characteristics and the magnetization-flop behavior in magnetic tunnel junctions exchange biased by synthetic antiferromagnets (SyAFs) are investigated by using a computer simulations based on a single-domain multilayer model. The bias field acting on the free layer is found to be sensitive to the thickness of neighboring layers, and the thickness dependence of the bias field is greater at smaller cell dimensions due to larger magnetostatic interactions. The resistance to magnetization flop increases with decreasing cell size due to increased shape anisotropy. When the cell dimensions are small and the synthetic antiferromagnet is weakly, or not pinned, the magnetization directions of the two layers sandwiching the insulating layer are aligned antiparallel due to a strong magnetostatic interaction, resulting in an abnormal magneto resistance (MR) change from the high-MR state to zero, irrespective of the direction of the free-layer switching. The threshold field for magnetization-flop is found to increase linearly with increasing antiferromagnetic exchange coupling in the synthetic antiferromagnet. Irrespective of the magnetic parameters and cell sizes, magnetization flop does not exist near zero applied field, indicating that magnetization flop is driven by the Zeeman energy.

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Physiological understanding of host-microbial pathogen interactions in the gut

  • Lee, Sei-Jung;Choi, Sang Ho;Han, Ho Jae
    • Korean Journal of Veterinary Research
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    • v.56 no.2
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    • pp.57-66
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    • 2016
  • The gut epithelial barrier, which is composed of the mucosal layer and the intestinal epithelium, has multiple defense mechanisms and interconnected regulatory mechanisms against enteric microbial pathogens. However, many bacterial pathogens have highly evolved infectious stratagems that manipulate mucin production, epithelial cell-cell junctions, cell death, and cell turnover to promote their replication and pathogenicity in the gut epithelial barrier. In this review, we focus on current knowledge about how bacterial pathogens regulate mucin levels to circumvent the epithelial mucus barrier and target cell-cell junctions to invade deeper tissues and increase their colonization. We also describe how bacterial pathogens manipulate various modes of epithelial cell death to facilitate bacterial dissemination and virulence effects. Finally, we discuss recent investigating how bacterial pathogens regulate epithelial cell turnover and intestinal stem cell populations to modulate intestinal epithelium homeostasis.

Role of Gap Junctions in the Endothelium-Dependent Hyperpolarization of Vascular Smooth Muscle Cells

  • Yamamoto, Yoshimichi;Klemm, Megan F.;Hashitani, Hikaru;Lang, Richard J.;Soji, Tsuyoshi;Suzuki, Hikaru
    • The Korean Journal of Physiology and Pharmacology
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    • v.5 no.1
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    • pp.1-8
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    • 2001
  • Hyperpolarization of arterial smooth muscle by acetylcholine is considered to be produced by the release of an unidentified chemical substance, an endothelium-derived hyperpolarizing factor (EDHF). Several chemicals have been proposed as the candidate for EDHF. However, none of them fulfil completely the nature and property of EDHF. Ultrastructural observation with electron microscope reveals that in some arteries, gap junctions are formed between endothelial and smooth muscle cells. In small arterioles, injection of gap junction permeable dyes into an endothelial cell results in a distribution of the dye to surrounding cells including smooth muscle cells. These observations allow the speculation that myoendothelial gap junctions may have a functional significance. Simultaneous measurement of the electrical responses in both endothelial and smooth muscle cells using the double patch clamp method demonstrates that these two cell types are indeed electrically coupled, indicating that they behave as a functional syncytium. The EDHF-induced hyperpolarization is produced by an activation of $Ca^{2+}-sensitive\;K^+-channels$ that are inhibited by charybdotoxin and apamin. Agonists that release EDHF increase $[Ca^{2+}]_i$ in endothelial cells but not in smooth muscle cells. Inhibition of gap junctions with chemical agents abolishes the agonist-induced hyperpolarization in smooth muscle cells but not in endothelial cells. All these observations can be explained if EDHF is an electrotonic signal propagating from endothelium to smooth muscle cells through gap junctions.

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Co-Silicide Device Characteristics in Embedded DRAM

  • Kim, Jong-Chae;Kim, Yeong-Cheol;Kim, Byung-Kook
    • Korean Journal of Crystallography
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    • v.12 no.3
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    • pp.162-165
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    • 2001
  • The EDL (Embedded DRAM and Logic) technologies with stack cell capacitors based on NO dielectric and Co-silicided source/drain junctions using a Ti capping material, were successfully implemented. The employed Co-silicided film exhibited junction leakage characteristics comparable to those of non-silicided junctions. Improved device characteristics without degradation of I/sub off/ was also achieved.

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A Freeze-fracture Study on the Odontoblast of Dental Pulp in the Rat Incisor (흰쥐 절치치수의 Odontoblast에 관한 Freeze-Fracture 연구)

  • Kim, Myung-Kook
    • Applied Microscopy
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    • v.16 no.2
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    • pp.1-13
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    • 1986
  • The purpose of this study was to investigate the morphology and intercellular junctions of the odontoblast of dental pulp in the rat incisor by means of the freeze fracture electron microscopy. Twenty male Sprague-Dawley rats weighing $150{\sim}200g$ were used. After being anesthetized by an intraperitoneal injection of 0.5 ml sodium pentobarbital per kg in body weight(60 mg/ml) the animals were perfused with 2.5% glutaraldehyde-2% paraformaldehyde fixative in 0.1 M cacodylate buffer, pH 7.2 through the ascending aorta for one hour. The incisors were carefully extracted from the jaws and demineralized by suspending them in 0.1 M EDTA in 3% glutaraldehyde (pH 7.2) for two weeks. After demineralization, the specimens were obtained from the portion divided into five equal parts. For freeze-fracture replication, demineralized tissues were infiltrated for several hours with 10%, 25% glycerol in 0.1M cacodylate buffer as a cryoprotectant and then frozen in liquid Freon 22 and stored in liquid nitrogen. Fracturing and replication were done in Balzers BAF 400D high-vacuum freeze-fracture apparatus at $-120^{\circ}C$ under routine $5X10^{-7}$ Torr vacuum. The tissue was immediately replicated with platinum unidirectionally at $45^{\circ}$ angle and reinforced with carbon at $90^{\circ}$ angle unidirectionally or by using a rotary stage. The replication process was monitored by a quartz-crystal device. The replicas were immersed in 100% methanol overnight. The tissue was then digested from the replica by clorox (laundry bleach), placed into 5% EDTA, and washed repeatedly with distilled water. The replicas were picked up on 0.3% formvar-coated 75 mesh grids and examined in the JEOL 100B electron microscope. The results were as follows; 1. Both in thin sections and freeze-fracture replicas, three types of intercellular junctions were recognizable in the plasma membrane of odontoblast: gap junction, tight junction and desmosome-like junction. 2. The nuclear pores were evenly distributed over the nuclear envelope. The pore complex formed a ring about 70 nm in diameter. 3. Gap junctions were found between odontoblasts as well as odontoblasts and neighbouring pulp cells (fibroblast, subodontoblastic cell process, nerve-like fibre). Gap junctions, which were round, ellipsoid and pear-shaped and 600 nm in diameter, were observed in the odontoblast. 4. Numerous round and ellipsoid gap junctions could be frequently seen on the plasma membranes in cell body and apical part of the odontoblasts. On the P face, the junctions were recognized as a cluster of closely packed particles, measuring about 9 nm in diameter, and on the E face, the junctions were recognized as a shallow grooves.

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Gilgyung-tang Inhibits the Migration and Invasion of Human Bladder Cancer 5637 Cells through the Tightening of Tight Junctions and Inhibition of Matrix Metalloproteinase Activity (길경탕의 치밀결합 강화 및 MMPs의 활성 억제를 통한 인체방광암세포의 이동성 및 침윤성의 억제)

  • Hong, Su-hyun;Choi, Yung-hyun
    • The Journal of Internal Korean Medicine
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    • v.37 no.1
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    • pp.16-25
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    • 2016
  • Objectives: Gilgyung-tang (GGT) has been used as one of the main multi-herb formulas to treat “Peo-ong” (lung abscess). In this study, we investigated the inhibitory effects of water extracts of GGT on cell migration and invasion, two critical cellular processes that are often deregulated during metastasis, in human bladder cancer 5637 cells.Methods: Effects on cell viability were quantified using an MTT assay. To analyze the anti-metastatic effects, we conducted a wound healing migration assay, an in vitro invasiveness assay, and a measurement of the transepithelial electrical resistance (TER). The expression of protein and mRNA were measured by Western blotting and real-time polymerase chain reaction (RT-PCR), respectively.Results: GGT markedly inhibited the cell motility and invasiveness of 5637 cells within the concentration range that was not cytotoxic. The inhibitory effects of GGT on cell invasiveness were associated with tightening of the tight junctions (TJs), which was demonstrated by an increase in the TER. The RT-PCR and Western blotting results indicated that GGT decreased the levels of claudin proteins. GGT also inhibited the activity and expression of matrix metalloproteinase (MMP)-2 and -9 and simultaneously increased the levels of tissue inhibitor of metalloproteinase-1 and -2.Conclusions: Our findings suggest that GGT reduces both the migration and the invasion of 5637 cells by modulating the activity of TJs and MMPs.

Low-Temperature Poly-Si TFT Charge Trap Flash Memory with Sputtered ONO and Schottky Junctions

  • An, Ho-Myoung;Kim, Jooyeon
    • Transactions on Electrical and Electronic Materials
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    • v.16 no.4
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    • pp.187-189
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    • 2015
  • A charge-trap flash (CTF) thin film transistor (TFT) memory is proposed at a low-temperature process (≤ 450℃). The memory cell consists of a sputtered oxide-nitride-oxide (ONO) gate dielectric and Schottky barrier (SB) source/drain (S/D) junctions using nickel silicide. These components enable the ultra-low-temperature process to be successfully achieved with the ONO gate stacks that have a substrate temperature of room temperature and S/D junctions that have an annealing temperature of 200℃. The silicidation process was optimized by measuring the electrical characteristics of the Ni-silicided Schottky diodes. As a result, the Ion/Ioff current ratio is about 1.4×105 and the subthreshold swing and field effect mobility are 0.42 V/dec and 14 cm2/V·s at a drain voltage of −1 V, respectively.

Fine structure of the intercalated disc and cardiac junctions in the black widow spider Latrodectus mactans

  • Yan Sun;Seung-Min Lee;Bon-Jin Ku;Myung-Jin Moon
    • Applied Microscopy
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    • v.50
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    • pp.20.1-20.9
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    • 2020
  • Arthropods have an open circulatory system with a simple tubular heart, so it has been estimated that the contractile pumping structure of the cardiac muscle will be less efficient than that of vertebrates. Nevertheless, certain arthropods are known to have far superior properties and characteristics than vertebrates, so we investigated the fine structural features of intercalated discs and cardiac junctions of cardiac muscle cells in the black widow spider Latrodectus mactans. Characteristically, the spider cardiac muscle has typical striated features and represents a functional syncytium that supports multiple connections to adjacent cells by intercalated discs. Histologically, the boundary lamina of each sarcolemma connects to the basement membrane to form an elastic sheath, and the extracellular matrix allows the cells to be anchored to other tissues. Since the intercalated disc is also part of sarcolemma, it contains gap junctions for depolarization and desmosomes that keep the fibers together during cardiac muscle contraction. Furthermore, fascia adherens and macula adherens (desmosomes) were also identified as cell junctions in both sarcolemma and intercalated discs. To enable the coordinated heartbeat of the cardiac muscle, the muscle fibers have neuronal innervations by multiple axons from the motor ganglion.