• Molecules and Cells
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• 제27권2호
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• pp.191-198
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• 2009

The present study was undertaken to determine whether treatment with genistein, the plant-derived estrogen-like compound influences agonist-induced vascular smooth muscle contraction and, if so, to investigate related mechanisms. The measurement of isometric contractions using a computerized data acquisition system was combined with molecular experiments. Genistein completely inhibited KCl-, phorbol ester-, phenylephrine-, fluoride- and thromboxane $A_2$-induced contractions. An inactive analogue, daidzein, completely inhibited only fluoride-induced contraction regardless of endothelial function, suggesting some difference between the mechanisms of RhoA/Rho-kinase activators such as fluoride and thromboxane $A_2$. Furthermore, genistein and daidzein each significantly decreased phosphorylation of MYPT1 at Thr855 had been induced by a thromboxane $A_2$ mimetic. Interestingly, iberiotoxin, a blocker of large-conductance calcium-activated potassium channels, did not inhibit the relaxation response to genistein or daidzein in denuded aortic rings precontracted with fluoride. In conclusion, genistein or daidzein elicit similar relaxing responses in fluoride-induced contractions, regardless of tyrosine kinase inhibition or endothelial function, and the relaxation caused by genistein or daidzein was not antagonized by large conductance $K_{Ca}$-channel inhibitors in the denuded muscle. This suggests that the RhoA/Rho-kinase pathway rather than $K^+$- channels are involved in the genistein-induced vasodilation. In addition, based on molecular and physiological results, only one vasoconstrictor fluoride seems to be a full RhoA/Rho-kinase activator; the others are partial activators.

• The Korean Journal of Physiology and Pharmacology
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• 제27권2호
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• pp.187-196
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• 2023

Transient receptor potential canonical (TRPC) channels are non-selective calcium-permeable cation channels. It is suggested that TRPC4β is regulated by phospholipase C (PLC) signaling and is especially maintained by phosphatidylinositol 4,5-bisphosphate (PIP₂). In this study, we present the regulation mechanism of the TRPC4 channel with PIP₂ hydrolysis which is mediated by a channel-bound PLCδ1 but not by the G_qPCR signaling pathway. Our electrophysiological recordings demonstrate that the Ca²⁺ via an open TRPC4 channel activates PLCδ1 in the physiological range, and it causes the decrease of current amplitude. The existence of PLCδ1 accelerated PIP₂ depletion when the channel was activated by an agonist. Interestingly, PLCδ1 mutants which have lost the ability to regulate PIP₂ level failed to reduce the TRPC4 current amplitude. Our results demonstrate that TRPC4 self-regulates its activity by allowing Ca²⁺ ions into the cell and promoting the PIP₂ hydrolyzing activity of PLCδ1.

• Molecules and Cells
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• 제21권1호
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• pp.52-62
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• 2006

In previous reports we demonstrated that ginsenosides, active ingredients of Panax ginseng, affect some subsets of voltage-dependent $Ca^{2+}$ channels in neuronal cells expressed in Xenopus laevis oocytes. However, the major component(s) of ginseng that affect cloned $Ca^{2+}$ channel subtypes such as ${\alpha}_{1C}$(L)-, ${\alpha}_{1B}$(N)-, ${\alpha}_{1A}$(P/Q)-, ${\alpha}_{1E}$(R)- and ${\alpha}_{1G}$(T) have not been identified. Here, we used the two-microelectrode voltage clamp technique to characterize the effects of ginsenosides and ginsenoside metabolites on $Ba^{2+}$ currents ($I_{Ba}$) in Xenopus oocytes expressing five different $Ca^{2+}$ channel subtypes. Exposure to ginseng total saponins (GTS) induced voltage-dependent, dose-dependent and reversible inhibition of the five channel subtypes, with particularly strong inhibition of the ${\alpha}_{1G}$-type. Of the various ginsenosides, $Rb_1$, Rc, Re, Rf, $Rg_1$, $Rg_3$, and $Rh_2$, ginsenoside $Rg_3$ also inhibited all five channel subtypes and ginsenoside $Rh_2$ had most effect on the ${\alpha}_{1C}$- and ${\alpha}_{1E}$-type $Ca^{2+}$ channels. Compound K (CK), a protopanaxadiol ginsenoside metabolite, strongly inhibited only the ${\alpha}_{1G}$-type of $Ca^{2+}$ channel, whereas M4, a protopanaxatriol ginsenoside metabolite, had almost no effect on any of the channels. $Rg_3$, $Rh_2$, and CK shifted the steady-state activation curves but not the inactivation curves in the depolarizing direction in the ${\alpha}_{1B}$- and ${\alpha}_{1A}$-types. These results reveal that $Rg_3$, $Rh_2$ and CK are the major inhibitors of $Ca^{2+}$ channels in Panax ginseng, and that they show some $Ca^{2+}$ channel selectivity.

• The Korean Journal of Physiology and Pharmacology
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• 제11권5호
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• pp.183-188
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• 2007

Using BHK cells with stable expression of cardiac $Na^+/Ca^{2+}$ exchanger(BHK-NCX1), reverse mode(i.e. $Ca^{2+}$ influx mode) of NCX1 current was recorded by whole-cell patch clamp. Repeated stimulation of reverse NCX1 produced a cytosolic $Ca^{2+}$-dependent long-term inactivation of the exchanger activity. The degrees of inactivation correlated with NCX1 densities of the cells and were attenuated by reduced $Ca^{2+}$ influx via the reverse exchanger. The inactivation of NCX1 was attenuated by(i) inhibition of $Ca^{2+}$ influx with reduced extracellular $Ca^{2+}$, (ii) treatment with NCX1 blocker($Na^{2+}$), and (iii) increase of cytoplasmic $Ca^{2+}$ buffer(EGTA). In BHK-NCX1 cells transiently expressing TRPV1 channels, $Ca^{2+}$ influx elicited by capsaicin produced a marked inactivation of NCX1. We suggest that cytoplasmic $Ca^{2+}$ has a dual effect on NCX1 activities, and that allosteric $Ca^{2+}$ activation of NCX1 can be opposed by the $Ca^{2+}$-dependent long-term inactivation in intact cells.

• Archives of Pharmacal Research
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• 제26권1호
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• pp.64-69
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• 2003

To analyze vasopressin-induced $Ca^{2+}$ increase in liver cells, rat hepatocytes were isolated and attached to collagen-coated cover slips. Using fura-2, a $Ca^{2+}$-sensing dye, changes in intracellular $Ca^{2+}$ concentration by vasopressin were monitored. Results in this communication suggested that vasopressin-induced $Ca^{2+}$ increase were composed of both $Ca^{2+}$ release from internal $Ca^{2+}$ stores and influx from the plasma membrane. The $Ca^{2+}$ influx consisted of two distinguishable components. One was dependent on the presence of vasopressin and the other was not. SK＆F96365 blocked vasopressin-induced $Ca^{2+}$ influx in a dose-dependent manner. Vasopressin-induced $Ca^{2+}$ release from internal stores diminished in a primary culture of hepatocytes according to the culture time. However, changes in vasopressin-induced $Ca^{2+}$ influx across the plasma membrane differed from changes in the $Ca^{2+}$ release from internal stores, suggesting two separate signalings from receptor activation to internal stores and to the plasma membrane.

• Molecules and Cells
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• 제37권9호
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• pp.656-663
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• 2014

Gintonin, a novel, ginseng-derived G protein-coupled lysophosphatidic acid (LPA) receptor ligand, elicits $[Ca^{2+}]_i$ transients in neuronal and non-neuronal cells via pertussis toxin-sensitive and pertussis toxin-insensitive G proteins. The slowly activating delayed rectifier $K^+$ ($I_{Ks}$) channel is a cardiac $K^+$ channel composed of KCNQ1 and KCNE1 subunits. The C terminus of the KCNQ1 channel protein has two calmodulin-binding sites that are involved in regulating $I_{Ks}$ channels. In this study, we investigated the molecular mechanisms of gintonin-mediated activation of human $I_{Ks}$ channel activity by expressing human $I_{Ks}$ channels in Xenopus oocytes. We found that gintonin enhances $I_{Ks}$ channel currents in concentration- and voltage-dependent manners. The $EC_{50}$ for the $I_{Ks}$ channel was $0.05{\pm}0.01{\mu}g/ml$. Gintonin-mediated activation 1 of the $I_{Ks}$ channels was blocked by an LPA1/3 receptor antagonist, an active phospholipase C inhibitor, an $IP_3$ receptor antagonist, and the calcium chelator BAPTA. Gintonin-mediated activation of both the $I_{Ks}$ channel was also blocked by the calmodulin (CaM) blocker calmidazolium. Mutations in the KCNQ1 $[Ca^{2+}]_i$/CaM-binding IQ motif sites (S373P, W392R, or R539W)blocked the action of gintonin on $I_{Ks}$ channel. However, gintonin had no effect on hERG $K^+$ channel activity. These results show that gintonin-mediated enhancement of $I_{Ks}$ channel currents is achieved through binding of the $[Ca^{2+}]_i$/CaM complex to the C terminus of KCNQ1 subunit.

• 한국통신학회논문지
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• 제33권12B호
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• pp.1050-1057
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• 2008

무선 메쉬 네트워크 (Wireless Mesh Network, WMN)는 최근 들어 무선 접속 네트워크에서 높은 네트워크 용량을 지원하기 위해 중요한 이슈로 대두되고 있다. WMN에서 주로 사용하는 네트워크 인터페이스 기술인 IEEE 802.11 표준에서는 IEEE 802.11b/g 또는 IEEE 802.11a와 같은 스펙에 따라 3개 또는 12개의 멀티채널을 지원한다. 하지만, WMN에서 채널 할당 알고리즘을 디자인 할 때 두 가지 중요한 문제, 즉 채널 의존성 문제와 채널 스캐닝 딜레이를 고려해야 한다. 채널 의존성 문제는 인터페이스의 동적인 채널 변경이 노드 연결성을 유지하기 위해 다른 인터페이스의 채널 변경을 일으키는 문제이다. 또한, 채널 스캐닝 밀레이는 인터페이스가 채널을 변경할 때마다 발생하며 성능에 영향을 미친다. 따라서, 본 논문에서는 멀티채널 WMN환경에서 이러한 문제점들을 위해 메쉬 게이트웨이로부터의 거리에 기반한 채널 할당 (Distance-Based Channel Assignment, DB-CA)을 제안한다. DB-CA에서는 서로 다른 채널을 사용하는 노드들이 통신을 하기 위해 채널 스캐닝을 수행하지 않고 오직 채널 스위칭만 수행하며, WMN의 메쉬 게이트웨이에 가까운 노드들이 사용하는 채널에 미칠 수 있는 간섭을 최소화 한다. 실험 결과를 통해 WMN에서 DB-CA가 기존 채널 할당 방식에 비해 향상된 성능을 나타냄을 확인하였다.

• 한국정보과학회논문지:컴퓨팅의 실제 및 레터
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• 제16권3호
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• pp.341-345
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• 2010

무선 메쉬 네트워크(Wireless Mesh Network, WMN)는 무선 링크를 이용한 무선 백본 네트워크를 형성하여 사용자에게 인터넷 서비스를 효율적으로 제공할 수 있는 해결책 중의 하나이다. WMN에서 널리 사용되고 있는 기술은 IEEE 802.11 이며, 이는 멀티 채널 (multichannel)과 멀티 레이트(multirate)를 제공한다. WMN에서 중요한 이슈 중의 하나는 네트워크 용량(capacity) 이며, 이를 높이기 위한 멀티 채널 프로토콜이 절실하다 하지만, 다른 레이트로 동작하는 무선 링크가 같은 채널에서 동작하면, 높은 레이트(high-rate) 링크의 성능이 낮은 레이트(low-rate) 링크로 인해 저하되는 성능 이상(performance anomaly) 현상이 발생한다. 이러한 문제를 완화하기 위해 본 논문에서는 데이터 레이트를 멀티채널로 분산시키기 위한 트리기반 채널 할당(Tree-based Channel Assignment, TreeCA) 프로토콜을 제안한다. TreeCA는 인터넷 트래픽을 효율적으로 전달하기 위해 트리 WMN 구조를 기반으로 채널 할당을 수행한다. 부모 노드는 다른 레이트로 연결된 자식 노드를 다른 채널로 분산시켜서, 성능 이상 현상을 완화한다. 시뮬레이션을 통해 TreeCA가 기존 멀티 채널 WMN 프로토콜 보다 향상된 성능을 보임을 관찰하였다.

• 한국응용약물학회:학술대회논문집
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• 한국응용약물학회 1998년도 Proceedings of UNESCO-internetwork Cooperative Regional Seminar and Workshop on Bioassay Guided Isolation of Bioactive Substances from Natural Products and Microbial Products
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• pp.148-149
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• 1998

It has been known that potassium channel openers are a new class of molecules that have attracted general interest because of their potent antihypertensive activity in vivo and vasorelaxant activity in vitro (Hamilton and Weston, 1989). In the present study, it was attempted to examine the effect of the potassium channel opener on catecholamine (CA) secretion evoked by cholinergic stimulation, membrane depolarization and calcium mobilization from the isolated perfused rat adrenal gland. The perfusion of pinacidil (30-300 uM) into an adrenal vein for 20 min produced relatively dose-dependent inhibition in CA secretion evoked by ACh (5.32 mM), high $K^{+}$ (56 mM), DMPP (100 uM for 2 min), McN-A-343 (100 uM for 2 min), cyclopiazonic acid (10 uM for 4 min) and Bay-K-8644 (10 uM for 4 min). Also, under the presence of minoxidil (100 uM), which is also known to be a potassium channel activator, CA secretory responses evoked by ACh, high potassium, DMPP, McN-A-343, Bay-K-8644 and cyclopiazonic acid were also significantly depressed. However, in adrenal glands preloaded with pinacidil (100 uM) under the presence of glibenclamide (1 uM), an antidiabetic sulfonylurea that has been shown to be a specific blocker of ATP-regulated potassium channels (for 20 min), CA secretory responses evoked by ACh, high potassium, DMPP, McN-A-343, Bay-K-8644 and cyclopiazonic acid were considerably recovered to a considerable extent of the normal release as compared to that of pinacidil only. These results, taken together, suggest that pinacidil cause the marked inhibition of CA secretion evoked by stimulation of cholinergic (both nicotinic and muscarinic) receptors as well as by membrane depolarization, indicating strongly that this effect may be mediated by inhibiting influx of extracellular calcium and release in intracellular calcium in the rat adrenomedullary chromaffin cells. Furthermore, these findings suggest strongly that these potassium channel openers-sensitive membrane potassium channels also play an important role in regulating CA secretion.

• 한국통신학회논문지
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• 제34권10B호
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• pp.964-972
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• 2009

무선 메쉬 네트워크(Wireless Mesh Network, WMN)는 다중 인터페이스와 다중 채널을 지원함으로써 기존의 무선 네트워크에서 발생되는 여러 문제점을 해결할 수 있는 핵심적인 미래기술로 대두되고 있다. WMN에서 다중 채널의 사용은 무선 네트워크 노드들이 사용하는 대역폭을 본질적으로 증가시켜준다. 그러나 이러한 대역폭을 충분히 사용하기 위해서는 다중 채널 환경에서 발생할 수 있는 채널 간 간섭을 최소화하는 채널 할당 기법이 요구된다. 본 논문에서는 WMN의 성능 향상을 위하여 클러스터링 기반의 최소 간섭 채널 할당(Minimum Interference Channel Assignment, MI-CA) 알고리즘을 제안한다. MI-CA 알고리즘은 클러스터 간의 Inter-Cluster 채널 할당과 클러스터 내부의 Intra-Cluster 채널 할당으로 구성된다. Inter-Cluster 채널 할당에서는 전체 WMN을 위한 베어본 채널을 MST(Minimum Spanning Tree) 기반으로 클러스터 헤더와 보더 노드에 할당하고, Intra-Cluster 채널 할당에서는 클러스터 멤버들 간에 직교채널을 재 할당함으로써 간섭을 최소화 한다. 시뮬레이션 결과 본 논문에서 제안한 MI-CA 알고리즘이 채널 간섭을 최소화함으로써 WMNs의 성능을 향상시킬 수 있음을 입증하였다.