• Title/Summary/Keyword: Saturable

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Characterization of Putrescine Uptake in Hamster Amelanocytic Melanoma AMEL-3 Cells

  • Garcia-Fernandez, Antonio J.;Rodriguez, Rosa A.;Perez-Pertejo, Yolanda;Balana-Fouce, Rafael
    • Molecules and Cells
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    • v.20 no.1
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    • pp.127-135
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    • 2005
  • The uptake of putrescine, spermidine and spermine by Fortner's hamster amelanocytic melanoma AMEL-3 cells was observed in this study to be time-dependent, temperature-sensitive, pH-dependent and saturable. Metabolic poisons nullified polyamine uptake, an indication that this is an energy-requiring mechanism. The presence of $Na^+$ ions was found to be requisite to full activity. Valinomycin, gramicidin, monensin and the calcium ionophore calcimycin were also observed to inhibit the process substantially. The transporter active site would seem to contain sulfhydryl groups. Other diamines and polyamine analogues, as well as cationic diamidines, suppressed putrescine uptake. The presence of the ornithine decarboxylase inhibitor DFMO in the culture medium induced putrescine inflows. Putrescine, in turn, induced the negative expression of the carrier, thus suggesting that this influx mechanism is governed by up/down regulation. The cationic diamidine CGP 40215A and its analogue CGP039937A competitively inhibited putrescine transport, with Ki values of 1.9 and $15{\mu}M$, respectively. The role of polyamine uptake in these cultures is discussed.

Helicobacter pylori Vacuolating Toxin Exhibits Polar Activity of $Cl^-$ Secretion and Secretory Response to Carbachol in T84 Cells

  • Jin, Nan-Ge;Jin, Yong-Ri;So, In-Suk;Kim, Ki-Whan
    • The Korean Journal of Physiology and Pharmacology
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    • v.8 no.5
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    • pp.289-293
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    • 2004
  • To investigate whether VacA (vacuolating toxin) produced by Helicobacter pylori Korean stain 99 induces intestinal secretion, purified VacA was added to T84 cell monolayers mounted in Ussing chambers, and electrical parameters were monitored. Mucosal addition of low pH-pretreated VacA increased short circuit current (Isc). The effect was time- and dose-dependent and saturable. The time-to-peak Isc was concentration-dependent. Chloride channel inhibitors, niflumic acid or 5-nitro-2-(3-phenylpropylamino)-benzoate (NPPB), inhibited VacA-stimulated Isc. Carbachol (CCh)-induced increase of Isc was prolonged by the addition of VacA to the mucosal side only. The effect was unaltered by the addition of niflumic acid. VacA did not show cytopathic effects. These studies indicate that VacA is a nonlethal toxin that acts in a polar manner on T84 monolayers to potentiate $Cl^-$ secretion and the response to CCh secretion without decrease in monolayer resistance. VacA may contribute to diarrhea diseases in human intestinal epithelial cells.

New ZVZCS PWM Converter For High Power Application (대전력 응용을 위한 새로운 ZVZCS PWM 컨버어터)

  • Ryoo, Hong-J.;Cho, Jung-G.;Yoo, Dong-W.;Rim, Geun-H.
    • Proceedings of the KIEE Conference
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    • 1996.07a
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    • pp.521-524
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    • 1996
  • A new zero voltage and nero current switching(ZVZCS) full bridge(FB) PWM converter b proposed to improve the performance of the previously presented ZVZCS-FB-PWM converters [7,8]. By adding a secondary active clamp and controlling the clamp switch moderately, ZVS(for leading-leg switches) are ZCS(for lagging-leg switches) are achieved without nay lossy components, the reverie avalanche break down of leading-leg IGBTs[7] or the saturable reactor in the primary[8]. Many advantages including simple circuit topology, high efficiency, and low cost mate the new converter attractive for high voltage and high power(> 10 kW) applications. The principle of operation is explained and analyzed. The features and design considerations of the new converter are also illustrated and verified on an 1.8 kW, 100 kHz IGBT based experimental circuit.

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Characterization of Binding of Treponema denticola to Immobilized Fibrinogen using the Fluorescent Fatty Acid Labeling Method

  • Hong, Jin;Lee, Si-Young
    • International Journal of Oral Biology
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    • v.35 no.3
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    • pp.107-111
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    • 2010
  • Treponema denticola is a gram-negative anaerobe that can cause periodontal disease. The adhesion of this bacterium to host tissues is considered to be the primary event in the colonization and infection of a host. Fibrinogen is generally found in damaged tissues resulting from periodontitis. The binding ability of T. denticola to fibrinogen may therefore be an important virulence factor in inducing periodontal diseases. It has been reported recently that oral spirochetes can be labeled with fluorescent fatty acids and we speculated that this labeling method could be used in an oral spirochete binding assay. The binding of several different strains of T. denticola to immobilized human fibrinogen was therefore tested using the fluorescent fatty acid labeling method. In the case of immobilized fibrinogen, the T. denticola ATCC 35405 strain showed saturable binding to immobilized fibrinogen. Indeed, all four different T. denticola strains tested in this experiment, T. denticola ATCC 35405, T. denticola ATCC 33520, T. denticola ATCC 35404 and T. denticola OTK showed binding to fibrinogen. The fluorescent fatty acid labeling method thus shows utility in binding assays for T. denticola, different strains of which can generally bind to immobilized fibrinogen.

Investigation of the Nature of the Endogenous Glucose Transporter(s) in Insect Cells

  • Lee, Chong-Kee
    • BMB Reports
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    • v.32 no.5
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    • pp.429-435
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    • 1999
  • Unlike the mammalian glucose transporter GLUT1, little is known about the nature of the endogenous sugar transporter(s) in insect cells. In order to establish the transport characteristics and other properties of the sugar transport proteins of Sf9 cells, a series of kinetic analyses was performed. A saturable transport system for hexose uptake has been revealed in the insect cells. The apparent affinity of this transport system(s) for 2-deoxy-D-glucose was relatively high, the $K_m$ for uptake being <0.5 mM. To further investigate the substrate and inhibitor recognition properties of the insect cell transporter, the ability of other sugars or drugs to inhibit 2-deoxy-D-glucose transport was examined by measuring inhibition constants ($K_j$). Transport was inhibited by D-mannose, D-glucose, and D-fructose. However, the apparent affinity of the C-4 epimer, D-galactose, for the Spodoptera transporter was relatively low, implying that the hydroxyl group at the C-4 position may play a role in the strong binding of glucose and mannose to the transporter. The results also showed that transport was stereoselective, being inhibited by D-glucose but not by L-glucose. It is therefore concluded that insect cells contain an endogenous glucose transport activity that in several aspects resembles the human erythrocyte glucose transporter. However, the mammalian and insect transporters were different in some of their kinetic properties, namely, their affinities for fructose and for cytochalasin B.

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Evidence for Sulfite Proton Symport in Saccharomyces cerevisiae

  • Park, Hoon;Alan T. Bakalinsky
    • Journal of Microbiology and Biotechnology
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    • v.14 no.5
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    • pp.967-971
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    • 2004
  • The kinetics of sulfite uptake were examined in a wild-type laboratory strain of Saccharomyces cerevisiae to determine if carrier-mediated sulfite uptake involved a proton symport, as previous studies on sulfite uptake have suggested both an active process and facilitated diffusion. Accumulation of intracellular sulfite was initially rapid and linear up to 50 sec. Uptake was saturable at final concentrations equal to or greater than 3 mM sulfite, and increased 2-fold in the presence of 2% glucose. Uptake was significantly reduced in cells pretreated with 100-500 $\mu$M carbonyl cyanide mchlorophenylhydrazone (CCCP) or 2,4-dinitrophenol (DNP), both of which dissipate proton gradients. Uptake was also significantly inhibited in the presence of 1 mM arsenate, an inhibitor of ATP synthesis. Extracellular alkalization was observed in cells incubated with 1-2 mM sulfite in a weak tartrate buffer at pH 3.5 and 4.5. These findings suggest that the bisulfite ion, $HSO_3^-$, an anionic form of sulfite, is taken up by a carrier-mediated proton symport. A met16 sull sul2 mutant, impaired in both sulfite formation and sulfate uptake, was found able to grow on a medium with sulfite as the sole Sulfur source, indicating that the sulfate transporters Sul1p and Sul2p are not required for sulfite uptake.

Characterization of Spermidine Transport System in a Cyanobacterium, Synechocystis sp. PCC 6803

  • Raksajit, Wuttinun;Yodsang, Panutda;Maenpaa, Pirkko;Incharoensakdi, Aran
    • Journal of Microbiology and Biotechnology
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    • v.19 no.5
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    • pp.447-454
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    • 2009
  • The transport of spermidine into a cyanobacterium, Synechocystis sp. pec 6803, was characterized by measuring the uptake of $^{14}C$-spermidine. Spermidine transport was shown to be saturable with an apparent affinity constant ($K_m$) value of $67{\mu}M$ and a maximal velocity ($V_{max}$) value of 0.45 nmol/min/mg protein. Spermidine uptake was pH-dependent with the pH optimum being 8.0. The competition experiment showed strong inhibition of spermidine uptake by putrescine and spermine, whereas amino acids were hardly inhibitory. The inhibition kinetics of spermidine transport by putrescine and spermine was found to be noncompetitive with $K_i$ values of 292 and $432{\mu}M$, respectively. The inhibition of spermidine transport by various metabolic inhibitors and ionophores suggests that spermidine uptake is energy-dependent. The diminution of cell growth was observed in cells grown at a high concentration of NaCl. Addition of a low concentration of spermidine at 0.5 mM relieved growth inhibition by salt stress. Upshift of the external osmolality generated by either NaCl or sorbitol caused an increased spermidine transport with about 30-40% increase at 10 mosmol/kg upshift.

The Torque Characteristics Analysis of the Single-Phase Switched Reluctance Motor According to the Starting Method (기동 방법에 따른 단상 SRM의 토크 특성 해석)

  • Kim, Jun-Ho;Kim, IL-Jung
    • Journal of the Korean Institute of Illuminating and Electrical Installation Engineers
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    • v.26 no.9
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    • pp.40-46
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    • 2012
  • The single-phase switched reluctance motor(SRM) has only one inductance variation and the positive torque is generated in the restricted section. So, it cannot be started by itself. To solve this problem, many researchers have addressed the several starting method for the single-phase SRM. This paper is focused on the torque characteristics of the single-phase SRM according to starting method. The four major starting method - permanent magnet, saturable stator pole, to grade the rotor, stepped rotor pole - is selected to analyze the torque characteristics. The analysis model of each starting method is designed to changed the pole shape or inserting other material in the basic model. The torque characteristics of each analysis model is obtained by using FEM analysis. The FEM analysis is performed at incremental rotor positions over half inductance cycle in any one pole with 250AT, 500AT, 750AT. The distortion factor of each analysis model is analyzed through the FFT to compare the distortion between basic model and four analysis model.

Choline and basic amine drugs efflux from brain to blood across the blood-brain barrier

  • Lee, Na-Young;Kang, Young-Sook
    • Proceedings of the Korean Society of Applied Pharmacology
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    • 2003.11a
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    • pp.107-107
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    • 2003
  • The purpose of this study is to examine that the efflux transport system for choline from brain to blood is present at the blood-brain barrier (BBB) using brain efflux index (BEI) method. [$^3$H]Choline was microinjected into parietal cortex area 2 (Par2) region of rat brain, and was eliminated from the brain with an apparent elimination half life of 45 min. The BBB efflux clearance of [$^3$H]choline was 0.12 $m\ell$/min/g brain, which was calculated from the efflux rate constant (1.5${\times}$10$\^$-2/ min$\^$-1/) and the distribution volume in the brain slice (8.1 $m\ell$/g brain). This process was saturable and significantly inhibited by various organic cationic compounds including hemicholinium-3, tetraethylammonium chloride (TEA) and verapamil, by antioxidant, ${\alpha}$-phenyl-n-tert-butyl nitrone (PBN), and by Alzheimer's disease therapeutics, such as acetyl $\ell$-carnitine and tacrine. In conclusion, this finding is the first direct in vivo evidence that choline is transported from brain to the blood across the BBB via a carrier-mediated efflux transport process.

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Modulation Depth Dependence of Timing Jitter and Amplitude Modulation in Mode-Locked Semiconductor Lasers (모드잠김 반도체 laser의 타이밍 지터및 크기 변조의 변조 신호 크기 의존성)

  • Kim, Ji-hoon;Bae, Seong-Ju;Lee, Yong-Tak
    • Proceedings of the Optical Society of Korea Conference
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    • 2000.02a
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    • pp.276.2-278
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    • 2000
  • In a recent years, a number of approaches have been studied, including passive, active, and hybrid mode-locking of semi-conductor lasers for short pulse generation and research has been devoted to achieve low timing-jitter operation since the timing jitter is unfavorable for system applications. Among the methods of mode locking, passive mode locking does not need external rf drives, and therefore the operation and fabrication procedures are simplified. In spite of these attractive advantages of passive mode-locked laser, it has critical drawbacks such as large timing jitter and the difficulty in synchronization with external circuits. Their inherent large timing jitter value was shown to be suppressed to certain levels by means of hybrid mode-locking technique$^{(1)}$ , where the saturable absorber section was modulated by an external signal with the cavity round trip frequency. Furthermore, the subharmonic mode-locking (SHML) technique alleviates the restrictions of high speed driving electronics. It has been demonstrated experimentally$^{(1)}$ that the hybrid subharmonic mode-locking technique has lead to significant reduction of the timing jitter. (omitted)

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