• Journal of Biomedical Engineering Research
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• v.43 no.5
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• pp.331-340
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• 2022

Early afterdepolarization (EAD), a significant cause of fatal ventricular arrhythmias including Torsade de Pointes (TdP) in long QT syndromes, is a depolarizing afterpotential at the plateau or repolarization phase in action potential (AP) profile early before completing one pace. AP duration prolongation is related to EAD but is not necessarily accounted for EAD. Several computational studies suggested EAD can form from an abnormality in the late plateau and/or repolarization phase of AP shape. In this sense, we hypothesized the slope during repolarization has the characteristics to predict TdP risk, mainly focusing on the maximum slope during repolarization (dVm/dt_{max_repol}). This study aimed to predict the sensitivity of dVm/dt_{max_repol} to ion channel conductances as a TdP risk metric through a population simulation considering multiple effects of simultaneous reduction in six ion channel conductances of g_NaL, g_Kr, g_Ks, g_to, g_K1, and g_CaL. Additionally, we verified the availability of dVm/dt_{max_repol} for TdP risk prediction through the correlation analysis with qNet, the representative TdP metric. We performed the population simulations based on the methodology of Gemmel et al. using the human ventricular myocyte model of Dutta et al. Among the sixion channel conductances, dVm/dt_{max_repol} and qNet responded most sensitively to the change in g_Kr, followed by g_NaL. Furthermore, dVm/dt_{max_repol} showed a statistically significant high negative correlation with qNet. The dVm/dt_{max_repol} values were significantly different according to three TdP risk levels of high, intermediate, and low by qNet (p<0.001). In conclusion, we suggested dVm/dt_{max_repol} as a new biomarker metric for TdP risk assessment.

• The Korean Journal of Physiology and Pharmacology
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• v.24 no.6
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• pp.529-543
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• 2020

In contrast to ventricular myocytes, the structural and functional importance of atrial transverse tubules (T-tubules) is not fully understood. Therefore, we investigated the ultrastructure of T-tubules of living rat atrial myocytes in comparison with ventricular myocytes. Nanoscale cell surface imaging by scanning ion conductance microscopy (SICM) was accompanied by confocal imaging of intracellular T-tubule network, and the effect of removal of T-tubules on atrial excitation-contraction coupling (EC-coupling) was observed. By SICM imaging, we classified atrial cell surface into 4 subtypes. About 38% of atrial myocytes had smooth cell surface with no clear T-tubule openings and intracellular T-tubules (smooth-type). In 33% of cells, we found a novel membrane nanostructure running in the direction of cell length and named it 'longitudinal fissures' (LFs-type). Interestingly, T-tubule openings were often found inside the LFs. About 17% of atrial cells resembled ventricular myocytes, but they had smaller T-tubule openings and a lower Z-groove ratio than the ventricle (ventricular-type). The remaining 12% of cells showed a mixed structure of each subtype (mixed-type). The LFs-, ventricular-, and mixed-type had an appreciable amount of reticular form of intracellular T-tubules. Formamide-induced detubulation effectively removed atrial T-tubules, which was confirmed by both confocal images and decreased cell capacitance. However, the LFs remained intact after detubulation. Detubulation reduced action potential duration and L-type Ca²⁺ channel (LTCC) density, and prolonged relaxation time of the myocytes. Taken together, we observed heterogeneity of rat atrial T-tubules and membranous ultrastructure, and the alteration of atrial EC-coupling by disruption of T-tubules.

• The Korean Journal of Physiology and Pharmacology
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• v.7 no.1
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• pp.25-28
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• 2003

Ion channel inhibitors are widely used for pharmacological discrimination between the different channel types as well as for determination of their functional role. In the present study, we tested the hypothesis that 4-aminopyridine (4-AP) could affect the large conductance $Ca^{2+}$-activated $K^+$ channel ($BK_{Ca}$) currents using perforated-patch or cell-attached configuration of patch-clamp technique in the rabbit pulmonary arterial smooth muscle. Application of 4-AP reversibly inhibited the spontaneous transient outward currents (STOCs). The reversal potential and the sensitivity to charybdotoxin indicated that the STOCs were due to the activation of $BK_{Ca}$. The $BK_{Ca}$ currents were recorded in single channel resolution under the cell-attached mode of patch-clamp technique for minimal perturbation of intracellular environment. Application of 4-AP also inhibited the single $BK_{Ca}$ currents reversibly and dose-dependently. The membrane potential of rabbit pulmonary arterial smooth muscle cells showed spontaneous transient hyperpolarizations (STHPs), presumably due to the STOC activities, which was also inhibited by 4-AP. These results suggest that 4-AP can inhibit $BK_{Ca}$ currentsin the intact rabbit vascular smooth muscle. The use of 4-AP as a selective voltage-dependent $K^+$ (KV) channel blocker in vascular smooth muscle, therefore, must be reevaluated.

• 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.

• JSTS:Journal of Semiconductor Technology and Science
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• v.8 no.3
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• pp.251-263
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• 2008

In this paper, we report an analytical modeling and 2-D Synopsys Sentaurus TCAD simulation of ion implanted silicon carbide MESFETs. The model has been developed to obtain the threshold voltage, drain-source current, intrinsic parameters such as, gate capacitance, drain-source resistance and transconductance considering different fabrication parameters such as ion dose, ion energy, ion range and annealing effect parameters. The model is useful in determining the ion implantation fabrication parameters from the optimization of the active implanted channel thickness for different ion doses resulting in the desired pinch off voltage needed for high drain current and high breakdown voltage. The drain current of approximately 10 A obtained from the analytical model agrees well with that of the Synopsys Sentaurus TCAD simulation and the breakdown voltage approximately 85 V obtained from the TCAD simulation agrees well with published experimental results. The gate-to-source capacitance and gate-to-drain capacitance, drain-source resistance and trans-conductance were studied to understand the device frequency response. Cut off and maximum frequencies of approximately 10 GHz and 29 GHz respectively were obtained from Sentaurus TCAD and verified by the Smith's chart.

• The Korean Journal of Physiology and Pharmacology
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• v.2 no.2
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• pp.133-145
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• 1998

$Ca^{2+}-signals$ in endothelial cells are determined by release from intracellular stores and entry through the plasma membrane. In this review, the nature of $Ca^{2+}$ entry and mechanisms of its control are reviewed. The following ion channels play a pivotal role in regulation of the driving force for $Ca^{2+}$ entry: an inwardly rectifying $K^+$ channel, identified as Kir2.1, a big-conductance, $Ca^{2+}-activated$ $K^+$ channel (hslo) and at least two $Cl^-$ channels (a volume regulated $Cl^-$ channel, VRAC, and a $Ca^{2+}$ activated $Cl^-$ channel, CaCC). At least two different types of $Ca^{2+}$-entry channels exist: 1. A typical CRAC-like, highly selective $Ca^{2+}$ channel is described. Current density for this $Ca^{2+}$ entry is approximately 0.1pA/pF at 0 mV and thus 10 times smaller than in Jurkat or mast cells. 2. Another entry pathway for $Ca^{2+}$ entry is a more non-selective channel, which might be regulated by intracellular $Ca^{2+}$. Although detected in endothelial cells, the functional role of trp1,3,4 as possible channel proteins is unclear. Expression of trp3 in macrovascular endothelial cells from bovine pulmonary artery induced non-selective cation channels which are probably not store operated or failed to induce any current. Several features as well as a characterisation of $Ca^{2+}$-oscillations in endothelial cells is also presented.

• Journal of Chest Surgery
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• v.18 no.1
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• pp.1-6
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• 1985

Isolated sinus node cells of the rabbit were used to assess the effects of extracellular Na, K, Ca and Mg concentrations on cardiac pacemaker activity. With intracellular glass micro-electrodes spontaneous action potentials of SA node were recorded and the effects of various ions and their blockers were analyzed in terms of the cycle length, the amplitude and the duration of action potentials, the results obtained were as follows. 1. Sodium reduction [up to 30%] decreased the amplitude of action potential and lengthened the cycle length. TTX, specific blocker of Na channel slightly lengthened the cycle length. 2. Increasing potassium ion concentration, the duration of action potential decreased and the frequency increased in 6mM, however, spontaneous action potential was stopped in 24 mM. Barium ion known to be decreasing K conductance increased the duration of action potential but no significant change in the cycle length was noticed. 3. Calcium ion has shortening effect on the duration and the cycle length of action potential but not with dose-dependent manner. Cadmium ion .[0.02mM] lengthened cycle length and the duration of action potential. 4. Increasing the concentration of magnesium ion the cycle length was lengthened, significantly.

• BMB Reports
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• v.37 no.4
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• pp.460-465
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• 2004

A 16-residue polypeptide model with the sequence acetyl-YALSLAATLLKEAASL-OH was derived by rational de novo peptide design. The designed sequence consists of amino acid residues with high propensity to adopt an alpha helical conformation, and sequential order was arranged to produce an amphipathic surface. The designed sequence was chemically synthesized using a solid-phase method and the polypeptide was purified by reverse-phase liquid chromatography. Molecular mass analysis by electro-spray ionization mass spectroscopy confirmed the correct designed sequence. Structural characterization by circular dichroism spectroscopy demonstrated that the peptide adopts the expected alpha helical conformation in 50% acetonitrile solution. Liposome binding assay using Small Unilamellar Vesicle (SUV) showed a marked release of entrapped glucose by interaction between the lipid membrane and the tested peptide. The channel-forming activity of the peptide was revealed by a planar lipid bilayer experiment. An analysis of the conducting current at various applied potentials suggested that the peptide forms a cationic ion channel with an intrinsic conductance of 188 pS. These results demonstrate that a simple rational de novo design can be successfully employed to create short peptides with desired structures and functions.

• The Korean Journal of Physiology and Pharmacology
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• v.7 no.5
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• pp.267-273
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• 2003

We have investigated the effect on inducing substate(s) of positively charged residues replaced in position 172 of the second transmembrane domain in murine inward rectifier potassium channels, formed by stable or transient transfection of Kir2.1 gene in MEL or CHO cells. Single channel recordings were obtained from either cell-attached patches or inside-out patches excised into solution containing 10 mM EDTA to rule out the effect of $Mg^{2+}$ on the channel gating. The substate(s) could be recorded with all mutants D172H, D172K and D172R. The unitary current-voltage (I-V) relation was not linear with D172H at $pH_i$ 6.3, whereas the unitary I-V relation was linear at $pH_i$ 8.0. The relative occupancy at $S_{LC}$ was increased from 0.018 at $pH_i$ 8.0 to 0.45 at $pH_i$ 5.5. In H-N dimer, that was increased from 0.016 at $pH_i$ 8.0 to 0.23 at $pH_i$ 5.5. The larger the size of the side chain or $pK_a$ with mutants (D172H, D172K and D172R), the more frequent the transitions between the fully open state and substate within an opening. The conductance of the substate also depended upon the pKa or the size of the side chain. The relative occupancy at substate $S_{LC}$ with monomer D172K (0.50) was less than that in K-H dimer (0.83). However, the relative occupancy at substate with D172R (0.79) was similar to that with R-N dimer (0.82). In the contrary to ROMK1, positive charge as well as negative charge in position 172 can induce the substate rather than block the pore in murine Kir2.1. The single channel properties of the mutant, that is, unitary I-V relation, the voltage dependence of the mean open time and relative occupancy of the substates and the increased latency to the first opening, explain the intrinsic gating observed in whole cell recordings.

• Journal of the Institute of Electronics Engineers of Korea SD
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• v.47 no.10
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• pp.14-22
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• 2010

The source/channel/drain regions are formed by ion implantation with different dopant types of $n^+/p^{(+)}/n^+$ in the fabrication of the conventional n-type metal-oxide-semiconductor field effect transistor(NMOSFET). In implementing the ultra-small devices with channel length of sub-30 nm, in order to achieve the designed effective channel length accurately, low thermal budget should be considered in the fabrication processes for minimizing the lateral diffusion of dopants although the implanted ions should be activated as completely as possible for higher on-current level. Junctionless (JL) MOSFETs fully capable of the the conventional NMOSFET operations without p-type channel for enlarging the process margin are under researches. In this paper, the optimum design of the JL MOSFET based on silicon nanowire (SNW) structure is carried out by 3-D device simulation and the basic radio frequency (RF) characteristics such as conductance, maximum oscillation frequency($f_{max}$), current gain cut-off frequency($f_T$) for the optimized device. The channel length was 30 run and the design variables were the channel doping concentration and SNW radius. For the optimally designed JL SNW NMOSFET, $f_T$ and $f_{max}$ high as 367.5 GHz and 602.5 GHz could be obtained, respectively, at the operating bias condition $V_{GS}$ = $V_{DS}$ = 1.0 V).