• KSBB Journal
• /
• v.10 no.1
• /
• pp.63-70
• /
• 1995

The effect of operating conditions on separation of soybean proteins in a home-made free-flow electrophoresis apparatus was investigated. Measurement of the pH, conductivity, and UV-absorbance(280 nm) were carried out at each run and the purity of the sample was tested with SDS-PAGE analysis. The soybean extract pretreated with Tris and boric acid was mixed with the amino acids composed of glutamic acid, histidine, arginine, glycine(1 mM each) with glycyl-glycine(2mM) and KCl(1mM). When the cellulose acetate was used as a compartment between the electrode and the buffer solution in the cell, pH distribution in the separation cell varied from 3.0 at the anodic side to 8.0 at the cathodic side and had two inflection point. The applied voltage was from 300V to 1000V and the separation was better at a higher voltage but the voltage was limited by the capability of the cooling system due to Joule heat. The proteins focused near the middle of the channel. From the change of pH and conductivity it was found that the ions in the channel moved out to the electrodes through the membrane. In the case when the concentration of the buffer solution was increased 5 times, proteins were focused at 300V. We could not increase up to the ten times of the concentration since the temperature difference between inlet and outlet was more than $25^{\circ}C$ and denaturation of proteins was expected. When ion-exchange membranes were used U-type pH distribution was set up due to the ionic polarization near the membrane. The commercial ampholytes, instead of the mixed amino acids showed not much improvements in purity of the separated sample.

• The Korean Journal of Physiology and Pharmacology
• /
• v.17 no.2
• /
• pp.127-132
• /
• 2013

Ginsenosides, one of the active ingredients of Panax ginseng, show various pharmacological and physiological effects, and they are converted into compound K (CK) or protopanaxatriol (M4) by intestinal microorganisms. CK is a metabolite derived from protopanaxadiol (PD) ginsenosides, whereas M4 is a metabolite derived from protopanaxatriol (PT) ginsenosides. The ${\gamma}$-aminobutyric acid $receptor_C$ ($GABA_C$) is primarily expressed in retinal bipolar cells and several regions of the brain. However, little is known of the effects of ginsenoside metabolites on $GABA_C$ receptor channel activity. In the present study, we examined the effects of CK and M4 on the activity of human recombinant $GABA_C$ receptor (${\rho}$ 1) channels expressed in Xenopus oocytes by using a 2-electrode voltage clamp technique. In oocytes expressing $GABA_C$ receptor cRNA, we found that CK or M4 alone had no effect in oocytes. However, co-application of either CK or M4 with GABA inhibited the GABA-induced inward peak current ($I_{GABA}$). Interestingly, pre-application of M4 inhibited $I_{GABA}$ more potently than CK in a dose- dependent and reversible manner. The half-inhibitory concentration ($IC_{50}$) values of CK and M4 were $52.1{\pm}2.3$ and $45.7{\pm}3.9{\mu}M$, respectively. Inhibition of $I_{GABA}$ by CK and M4 was voltage-independent and non-competitive. This study implies that ginsenoside metabolites may regulate $GABA_C$ receptor channel activity in the brain, including in the eyes.

• The Korean Journal of Physiology and Pharmacology
• /
• v.3 no.1
• /
• pp.1-10
• /
• 1999

We sought to find out the mechanism of vascular relaxation by extracellular $K^+$ concentration $([K^+]_o)$ in the cerebral resistant arteriole from rabbit. Single cells were isolated from the cerebral resistant arteriole, and using voltage-clamp technique barium-sensitive $K^+$ currents were recorded, and their characteristics were observed. Afterwards, the changes in membrane potential and currents through the membrane caused by the change in $[K^+]_o$ was observed. In the smooth muscle cells of cerebral resistant arteriole, ion currents that are blocked by barium, 4-aminopyridine (4-AP), and tetraethylammonium (TEA) exist. Currents that were blocked by barium showed inward rectification. When the $[K^+]_o$ were 6, 20, 60, and 140 mM, the reversal potentials were $-82.7{\pm}1.0,\;-49.5{\pm}1.86,\;-26{\pm}1.14,\;-5.18{\pm}1.17$ mV, respectively, and these values were almost identical to the calculated $K^+$ equilibrium potential. The inhibition of barium-sensitive inward currents by barium depended on the membrane potential. At the membrane potentials of -140, -100, and -60 mV, $K_d$ values were 0.44, 1.19, and 4.82 ${\mu}M,$ respectively. When $[K^+]_o$ was elevatedfrom 6 mM to 15 mM, membrane potential hyperpolarized to -50 mV from -40 mV. Hyperpolarization by $K^+$ was inhibited by barium but not by ouabain. When the membrane potential was held at resting membrane potential and the $[K^+]_o$ was elevated from 6 mM to 15 mM, outward currents increased; when elevated to 25 mM, inward currents increased. Fixing the membrane potential at resting membrane potential and comparing the barium-sensitive outward currents at $[K^+]_o$ of 6 and 15 mM showed that the barium- sensitive outward current increased at 15 mM $K^+.$ From the above results the following were concluded. Barium-sensitive $K^+$?channel activity increased when $[K^+]_o$ is elevated and this leads to an increase in $K^+-outward$ current. Consequently, the membrane potential hyperpolarizes, leading to the relaxation of resistant arteries, and this is thought to contribute to an increase in the local blood flow of brain.

• The Korean Journal of Physiology and Pharmacology
• /
• v.26 no.4
• /
• pp.277-285
• /
• 2022

To investigate the adverse effects of clozapine on cardiovascular ion channels, we examined the inhibitory effect of clozapine on voltage-dependent K⁺ (Kv) channels in rabbit coronary arterial smooth muscle cells. Clozapine-induced inhibition of Kv channels occurred in a concentration-dependent manner with an half-inhibitory concentration value of 7.84 ± 4.86 µM and a Hill coefficient of 0.47 ± 0.06. Clozapine did not shift the steady-state activation or inactivation curves, suggesting that it inhibited Kv channels regardless of gating properties. Application of train pulses (1 and 2 Hz) progressively augmented the clozapine-induced inhibition of Kv channels in the presence of the drug. Furthermore, the recovery time constant from inactivation was increased in the presence of clozapine, suggesting that clozapine-induced inhibition of Kv channels is use (state)-dependent. Pretreatment of a Kv1.5 subtype inhibitor decreased the Kv current amplitudes, but additional application of clozapine did not further inhibit the Kv current. Pretreatment with Kv2.1 or Kv7 subtype inhibitors partially blocked the inhibitory effect of clozapine. Based on these results, we conclude that clozapine inhibits arterial Kv channels in a concentration-and use (state)-dependent manner. Kv1.5 is the major subtype involved in clozapine-induced inhibition of Kv channels, and Kv2.1 and Kv7 subtypes are partially involved.

• The Korean Journal of Physiology and Pharmacology
• /
• v.27 no.6
• /
• pp.521-531
• /
• 2023

Transmembrane protein TMEM16A, which encodes calcium-activated chloride channel has been implicated in tumorigenesis. Overexpression of TMEM16A is associated with poor prognosis and low overall survival in multiple cancers including lung adenocarcinoma, making it a promising biomarker and therapeutic target. In this study, three structure-related sesquiterpene lactones (mecheliolide, costunolide and dehydrocostus lactone) were extracted from the traditional Chinese medicine Aucklandiae Radix and identified as novel TMEM16A inhibitors with comparable inhibitory effects. Their effects on the proliferation and migration of lung adenocarcinoma cells were examined. Whole-cell patch clamp experiments showed that these sesquiterpene lactones potently inhibited recombinant TMEM16A currents in a concentration-dependent manner. The half-maximal concentration (IC50) values for three tested sesquiterpene lactones were 29.9 ± 1.1 µM, 19.7 ± 0.4 µM, and 24.5 ± 2.1 µM, while the maximal effect (E_max) values were 100.0% ± 2.8%, 85.8% ± 0.9%, and 88.3% ± 4.6%, respectively. These sesquiterpene lactones also significantly inhibited the endogenous TMEM16A currents and proliferation, and migration of LA795 lung cancer cells. These results demonstrate that mecheliolide, costunolide and dehydrocostus lactone are novel TMEM16A inhibitors and potential candidates for lung adenocarcinoma therapy.

• Proceedings of the KIEE Conference
• /
• 1995.07c
• /
• pp.1085-1087
• /
• 1995

A large leakage current may be one of the critical issues for poly-silicon thin film transistors(poly-Si TFTs) for LCD applications. In order to reduce the leakage current of poly-Si TFTs, several offset gated structures have been reported. However, those devices, where the offset length in the source region is not same as that in the drain region, exhibit the asymmetric electrical performances such as the threshold voltage shift and the variation of the subthreshold slope. The different offset length is caused by the additional mask step for the conventional offset structures. Also the self-aligned implantation may not be applicable due to the mis-alignment problem. In this paper, we propose a new fabrication method for poly-Si TFTs with a self-aligned offset gated structure by employing a photo resistor reflow process. Compared with the conventional poly-Si TFTs, the device is consist of two gate electrodes, of which one is the entitled main gate where the gate bias is employed and the other is the entitled subgate which is separate from both sides of the main gate. The poly-Si channel layer below the offset oxide is protected from the injected ion impurities for the source/drain implantation and acts as an offset region of the proposed device. The key feature of our new device is the offset lesion due to the offset oxide. Our experimental results show that the offset region, due to the photo resistor reflow process, has been successfully obtained in order to fabricate the offset gated poly-Si TFTs. The advantages of the proposed device are that the offset length in the source region is the same as that in the drain region because of the self-aligned implantation and the proposed device does not require any additional mask process step.

• Transactions of the Korean Society of Mechanical Engineers B
• /
• v.36 no.6
• /
• pp.625-632
• /
• 2012

The objective of this paper is to demonstrate the cathode channel flooding effects at different stoichiometries in proton exchange membrane (PEM) fuel cells by using visualization techniques. The phenomena of liquid water formation and removal caused by current variations were also examined experimentally. Tests were conducted at cathode stoichiometries of 1.5 and 2.0, and the anode stoichiometry was fixed at 1.5. It is found that at an air-side stoichiometry of 2.0, liquid water begins to form and the flooding occurs faster than at an air-side stoichiometry of 1.5. Also, when the air-side stoichiometry of 1.5 is maintained, the dry-out phenomena is observed in the dry-out area 7.8 A following the field of flooding. Thus, a stoichiometry of 1.5 produced better performance in terms of membrane electrode assembly (MEA) durability and hydrogen ion conductivity than did a stoichiometry of 2.0, in which dry-out occurs beyond 8A.

• Journal of Physiology & Pathology in Korean Medicine
• /
• v.25 no.6
• /
• pp.996-999
• /
• 2011

This experiment was performed to investigate whether the combined-preparation of crude drugs (The $Well^{TM}$ Preparation, TW), has hypnotic effects and/or enhances pentobarbital-induced sleep behaviors. TW was mixed with water extracts of Ginseng Radix red, Germinated brown rice, cultured mountain ginseng, and 50% ethanol extracts of Longanae Arillus, Nelumbinis Folium and Chrysanthemi Flos. TW (100 mg/kg, p.o.) reduced sleep onset and prolonged sleep time induced by pentobarbital similar to muscimol (0.2 ${\mu}M$), a $GABA_A$ receptor agonist. Also, TW (2 ${\mu}g$/ml) and pentobarbital (2.5 ${\mu}M$) did not affect the chloride influx in primary cultured cerebellar granule cells, respectively, but the combined-treatment of TW (2 ${\mu}g$/ml) and pentobarbital (2.5 ${\mu}M$) increased the chloride influx onto the cells. In conclusion, TW augments pentobarbital-induced sleep behaviors; these effects may result from chloride channel activation.

• Journal of Ginseng Research
• /
• v.39 no.3
• /
• pp.257-264
• /
• 2015

Background: Korean Red Ginseng-a steamed root of Panax ginseng Meyer-has long been used as a traditional medicine in Asian countries. Its antipruritic effect was recently found, but no molecular mechanisms were revealed. Thus, the current study focused on determining the underlying molecular mechanism of Korean Red Ginseng extract (RGE) against histamine-induced itch at the peripheral sensory neuronal level. Methods: To examine the antipruritic effect of RGE, we performed in vivo scratching behavior test in mice, as well as in vitro calcium imaging and whole-cell patch clamp experiments to elucidate underlying molecular mechanisms. Results: The results of our in vivo study confirmed that RGE indeed has an antipruritic effect on histamine-induced scratching in mice. In addition, RGE showed a significant inhibitory effect on histamine-induced responses in primary cultures of mouse dorsal root ganglia, suggesting that RGE has a direct inhibitory effect on sensory neuronal level. Results of further experiments showed that RGE inhibits histamine-induced responses on cells expressing both histamine receptor subtype 1 and TRPV1 ion channel, indicating that RGE blocks the histamine receptor type 1/TRPV1 pathway in sensory neurons, which is responsible for histamine-dependent itch sensation. Conclusion: The current study found for the first time that RGE effectively blocks histamine-induced itch in peripheral sensory neurons. We believe that the current results will provide an insight on itch transmission and will be helpful in understanding how RGE exerts its antipruritic effects.

• Development and Reproduction
• /
• v.4 no.2
• /
• pp.243-250
• /
• 2000

To find the mechanism underlying the ADP-induced increase in the outward current in ovulated mouse oocytes, we examined changes in voltage-dependent currents using the whole cell voltage clamp technique and the internal perfusion technique. Eggs were collected from the oviduct of superovulated mice with PMSG and hCG. Membrane potential was held at -60 mV (or -80 mV in the case of recording $Ca^{2＋}$ currents) and step depolarizations or hyperpolarizations were applied for 300 ms. By step depolarizations, outward currents comprising steady-state and time-dependent components were elicited. They were generated in response to the positive potential more than 20 mV with severe outward rectification and were blocked by external TEA, a specific $K^{＋}$ channel blocker, suggesting that they be carried via $K^{＋}$ channels. Internally-perused 5 mM ADP gradually increased outward $K^{＋}$ currents (IK) 1 min after perfusion of ADP and reached slowly to maximum (150~170％) 5 min later over the positive potential range, implying that ADP might not be acted directly to the $K^{＋}$ channels. IK were decreased by 5 mM ATP without affecting the steady-state component of outward current. In contrast to the effect of ADP and ATP on IK, both effect of ATP and ADP on inward $Ca^{2＋}$ currents (ICa) could not be detected due to the continuous decrease in current amplitudes with time-lapse ("run-down" phenomena). To check if there is a G protein-involved regulation in the ionic current of mouse oocytes, 1 mM GTP was applied to the cytoplasmic side, and the outward current and inward currents were recorded. ICa was promptly increased in the presence of GTP whereas IK was not changed. from these results, it is concluded that the ATP-dependent regulation is likely linked in the ADP-induced increase in the outward $K^{＋}$ current, and G protein-involved cellular signalling might affect ion channels carrying $Ca^{2＋}$ and $K^{＋}$ in mouse oocytes.