• Journal of the Korean Institute of Electrical and Electronic Material Engineers
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• v.18 no.8
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• pp.708-713
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• 2005

An alumina membrane with nano-sized pore array by anodic oxidation using the thin film aluminum deposited on silicon wafer was fabricated. It Is important that the sample prepared by metal deposition method has a flat aluminum surface and a good adhesion between the silicon wafer and the thin film aluminum. The oxidation time was controlled by observation of current variation. While the oxalic acid with 0.2 M was used for low voltage anodization under 100 V, the chromic acid with 0.1 M was used for high voltage anodization over 100 V. The nano-sized pores with diameter of $60\~120$ nm was obtained by low voltage anodization of $40\~80$ V and those of $200\~300$ nm was obtained by high voltage anodization of $140\~200$ V. The pore widening process was employed for obtaining the one-channel with flat surface because the pores of the alumina membrane prepared by the fixed voltage method shows the structure of two-channel with rough surface. Finally, the sample was immersed to the phosphoric acid with 0.1 M concentration to etching the barrier layer.

• The Korean Journal of Physiology and Pharmacology
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• v.8 no.5
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• pp.259-264
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• 2004

Cytolysin produced by Vibrio vulnificus has been incriminated as one of the important virulence determinants in V. vulnificus infection. Ion selectivity of cytolysin-induced pores was examined in a CPAE cell, a cell line of pulmonary endothelial cell, using inside-out patch clamp techniques. In symmetrical NaCl concentration (140 mM), intracellular or extracellular application of cytolysin formed ion-permeable pores with a single channel conductance of $37.5{\pm}4.0$ pS. The pore currents were consistently maintained after washout of cytolysin. Replacement of $Na^+$ in bath solution with monovalent ions $(K^+,\;Cs^+\;or\;TEA^+)$ or with divalent ions $(Mg^{2+},\;Ca^{2+})$ did not affect the pore currents. When the NaCl concentration in bath solution was lowered from 140 to 60 and 20 mM, the reversal potential shifted from 0 to -11.8 and -28.2 mV, respectively. The relative permeability of the cytolysin pores to anions measured at $-40\;mV\;was\;Cl^-\;=\;NO_2^-\;{\geq}\;Br^-\;=\;I^-\;> \;SCN^-\;>\;acetate^-\;>\;isethionate^-\;>\;ascorbic acid^-\;>\;EDTA^{2-},$ in descending order. The cytolysin-induced pore current was blocked by $CI^-$ channel blockers or nucleotides. These results indicate that V. vulnificus cytolysin forms anion-selective pores in CPAE cells.

• Journal of Embryo Transfer
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• v.26 no.3
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• pp.209-214
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• 2011

Endometrium undergoing hormonal change plays important roles in preparation for implantation, fetal growth, and well-being. During pregnancy, cellular remodeling and hormonal changes in endometrium could change two-pore domain K$^+$ channel (K$_{2P}$) expression. This study was performed to identify whether K$_{2P}$ channel expression is changed in endometrium of pregnant Korean cattle, and whether the expression level is modulated by progesterone treatment. We investigated changes in the mRNA and protein expressions of K$_{2P}$ channel in pregnant endometrium using RT-PCR and Western blot analyses. The expression levels of all K$_{2P}$ channel mRNAs tested in this study, except that of TREK-1, were changed in the pregnant endometrium. mRNA levels of TASK-3 and TRAAK were significantly down-regulated, whereas those of TREK-2 and TRESK were up-regulated in the pregnant endometrium. In parallel with the RT-PCR results, Western blot analysis revealed up-regulations of TREK-2 (7.9-fold) and TRESK (2-fold) proteins levels in the pregnant endometrium. In addition, TREK-2 and TRESK protein levels were up-regulated in bovine endometrial cells by progesterone treatment (10 ${\mu}g$/ml). From these results, we suggest that the up-regulation of TREK-2 and TRESK by progesterone may contribute to the regulation of physiological changes during pregnancy.

• Parasites, Hosts and Diseases
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• v.41 no.2
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• pp.129-133
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• 2003

A complete cDNA sequence encoding a pore-forming subunit (Kir6.2) of ATP-senstive potassium channel in the adult worm, Clonorchis sinensis, termed CsKir6.2, was isolated from an adult CDNA library. The cDNA contained a single open-reading frame of 333 amino acids, which has a structural motif (a GFG-motif) of the putative pore-forming loop of the Kir6.2. Peculiarly, the Cskir6.2 shows a lack-sequence structure, which deleted 57 amino acids were deleted from its N-terminus. The predicted amino acid sequence revealed a highly conserved sequence as other known other Kir6.2 subunits. The mRNA was weekly expressed in the adult worm.

• Journal of Institute of Control, Robotics and Systems
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• v.14 no.5
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• pp.420-425
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• 2008

PMMA light guiding plate with nano-sized pattern was fabricated using anodized aluminum oxide membrane as a template for nano imprint lithography. Nano-sized pore arrays were prepared by the self-organization processes of the anodic oxidation using the aluminum plate with 99.999% purity. Since the aluminum plate has a rough surface, the aluminum plate with thickness of 1mm was anodized after the pre-treatments of chemical polishing, and electrochemical polishing. The surface morphology of the alumina obtained by the first anodization process was controlled by the concentration of electrochemical solution during the first anodization. The surface morphology of the alumina was also changed according to temperature of the solution during chemical polishing performed after first anodization. The pore widening process was employed for obtaining the one-channel with flat surface and height of the channel because the pores of the alumina membrane prepared by the fixed voltage method shows the structure of two-channel with rough surface. It is shown from SPM results that the nano-sized pattern on PMMA light guiding plate fabricated by nano imprint lithography method was well transferred from that of anodized aluminum oxide template.

• Membrane Journal
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• v.7 no.3
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• pp.142-149
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• 1997

The extended analysis on the diverging flow through asymmetric membrane pores has been performed in this study. Afore rigorous equations of velocity profile relevant to the divergent slit and cone shaped channels, which are widely used as a general pore model, have been obtained by employing a creeping flow approach of Newtonian fluids. As a degree of asymmetry (i.e., diverging angle) is increased, the predicted flow function shifts Toward the center region due to the incorporated wall effect, so that the overall velocity profile becomes decreased. It is true, as expected, that when the divergent channel is in the low diverging angle limit, the channel flow results in the Poiseuillean fashion by utilizing a lubrication approximation. The flow rate equation of each type of channel has been developed from the combined solution of velocity profile and pressure fields. The effect of diverging flow on the flow rate enhancement has been remarkably predicted, in which the flow rate increases with the increase of pore asymmetry. The advantage of our theoretical results lies in the analytical expression for the diverging flow behavior through pore channels as well as its ability to play a fundamental role on the related membrane filtrations such as microfiltration and ultrafiltration.

• Korean Journal of Materials Research
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• v.13 no.9
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• pp.593-597
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• 2003

Anodic aluminum oxide (AAO) membrane was made of aluminum sheet (99.6%, 0.2 mm thickness). The regular array of hexagonal nano pores or channels were prepared by two step anodization process. A detail description of the AAO fabrication is presented. After the 1st anodization in oxalic acid (0.3 M) at 45 V, The formed AAO was removed by etching in a solution of 6 wt% $H_3$$PO_4$＋1.8 wt% $H_2$$CrO_4$. The regular arrangement of the pores was obtained by the 2nd anodization, which was carried out in the same condition as the 1st anodization. Subsequently, the alumina barrier layer at the bottom of the channel layer was removed in phosphoric acid (1M) after removing of aluminum. Pore diameter, density, and thickness could be controlled by the anodization process parameters such as applied voltage, anodizing time, pore widening time, etc. The pore diameter is proportional to the applied voltage and pore widening time. The pore density and thickness can be controlled by anodization temperature and voltage.

• Journal of the Korea Academia-Industrial cooperation Society
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• v.12 no.6
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• pp.2660-2667
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• 2011

TREK-1 channel is a member of the two-pore domain potassium (K2P) channel family that is regulated by intracellular pH, membrane stretch, polyunsaturated fatty acids, temperature, and some neuroprotectant agents. TREK-1 channel can influence neuronal excitability by regulating leakage of potassium ions and resting membrane potential. TREK-1 channel has been shown to be overexpressed in prostate cancer cells. Although the importance of these properties, relatively little is known about flavonoid effects in the regulations of TREK-1 channel. The purpose of the study was to screening of flavonoids as the TREK-1 channel modulator using one of electrophysiological techniques such as excised inside-out patch configuration. We demonstrated blocking effect on TREK-1 channel by flavonoids such as epigallocatechin-3-gallate (EGCG), curcumin and quercetin in CHO cells transiently expressing TREK-1 channel. The inhibition of TREK-1 channel by quercetin and curcumin was reversible, whereas EGCG was little reversible. Quercetin, EGCG and curcumin decreased the relative channel activity to 73%, 91% and 94%, respectively. The half-inhibitory concentration (IC50) of curcumin, quercetin and EGCG was $1.04{\pm}0.19\;{\mu}M$, $1.13{\pm}0.26\;{\mu}M$ and $13.5{\pm}2.20\;{\mu}M$ in CHO cells expressing TREK-1 channel, respectively. These results indicate that flavonoids might regulate TREK-1 and this regulation might be one of the pharmacological actions of flavonoid in nervous systems and cancer cells.

• Journal of the Korea Academia-Industrial cooperation Society
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• v.17 no.3
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• pp.127-135
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• 2016

Two-pore domain potassium (K2P) channels are the targets of physiological stimuli, such as intracellular pH, bioactive lipids, and neurotransmitters, and they set the resting membrane potential. Some types of K2P channels play a critical role in both apoptosis and tumoriogenesis. Among the K2P channels, no antagonists of the TREK2 channel have been reported. The aim of the present study was to determine if the TREK2 channel is blocked and whether cell proliferation is influenced by flavonoids in the TREK2 overexpressing HEK293 cells (HEKT2). The electrophysiological current was recorded using single channel patch clamp techniques and cell proliferation was measured using a XTT assay. The electrophysiological results showed that the TREK2 channel activity was reduced to $91.5{\pm}13.1%$ (n=5) and $82.2{\pm}13.7%$ (n=5) by flavonoids, such as epigallocatechin-3-gallate (EGCG) and quercetin in HEKT2 cells, respectively. In contrast, the EGCG analogue, epicatechin (EC), had no significant inhibitory effects on the TREK2 single channel activity. In addition, cell proliferation was reduced to $69.4{\pm}14.0%$ (n=4) by ECGG in the HEKT2 cells. From these results, EGCG and quercetin represent the first known TREK2 channel inhibitors and only EGCG reduced HEKT2 cell proliferation. This suggests that the flavonoids may work primarily by inhibiting the TREK2 channel, leading to a change in the resting membrane potential, and triggering the initiation of a change in intracellular signaling for cell proliferation. TREK2 channel may, at least in part, contribute to cell proliferation.

• Journal of Embryo Transfer
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• v.24 no.3
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• pp.189-197
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• 2009

Two-pore domain $K^+(K_{2P})$ channels contribute to setting the resting membrane potential in excitable and nonexcitable cells. However, the cellular or tissue distribution and function of $K_{2P}$ channels expressed in mammalian germ cells and reproductive organs have not yet been reviewed by researchers. In this review, we focus on expression, localization and expected properties of $K_{2P}$ channels in germ cells and reproductive organs. The $K_{2P}$ channels are expressed in human cytotrophoblast cells, myometrium, placental vascular system, uterine smooth muscle, and pregnant term tissue, suggesting that $K_{2P}$ channels might be involved in the processes of pregnance. The $K_{2P}$ channels are also expressed in mouse zygotes, monkey sperm, ovary, testis, germ cells, and embryos of Korean cattle. Interestingly, $K_{2P}$ channels are modulated by changes in temperature and oxygen concentration which play an important role in embryonic development. Also, $K_{2P}$ channels are responsible for $K^+$ efflux during apoptotic volume decreases in mouse zygotes. These expression patterns and properties of the $K_{2P}$ channels in reproductive organs and germ cells are likely to help the understanding of ion channel-related function in reproductive physiology.