• The Korean Journal of Physiology and Pharmacology
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• 제4권6호
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• pp.445-453
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• 2000

The present investigation tested the hypothesis that the activation of protein kinase G (PKG) leads to a phosphorylation of $Ca^{2+}-activated$ potassium channel $(K_{Ca}\;channel)$ and is involved in the activation of $K_{Ca}$ channel activity in cerebral arterial smooth muscle cells of the rabbit. Single-channel currents were recorded in cell-attached and inside-out patch configurations of patch-clamp techniques. Both molsidomine derivative 3-morpholinosydnonimine-N-ethylcarbamide $(SIN-1,\;50\;{\mu}M)$ and 8-(4-Chlorophenylthio)-guanosine-3',5'-cyclic monophosphate $(8-pCPT-cGMP,\;100\;{\mu}M),$ a membrane-permeable analogue of cGMP, increased the $K_{Ca}$ channel activity in the cell-attached patch configuration, and the effect was removed upon washout of the drugs. In inside-out patches, single-channel current amplitude was not changed by SIN-1 and 8-pCPT-cGMP. Application of ATP $(100\;{\mu}M),$ cGMP $(100\;{\mu}M),$ ATP+cGMP $(100\;{\mu}M\;each),$ PKG $(5\;U/{\mu}l),$ ATP $(100\;{\mu}M)+PKG\;(5\;U/{\mu}l),$ or cGMP $(100\;{\mu}M)+PKG\;(5\;U/{\mu}l)$ did not increase the channel activity. ATP $(100\;{\mu}M)+cGMP\;(100\;{\mu}M)+PKG\;(5\;U/{\mu}l)$ added directly to the intracellular phase of inside-out patches increased the channel activity with no changes in the conductance. The heat-inactivated PKG had no effect on the channel activity, and the effect of PKG was inhibited by 8-(4-Chlorophenylthio)-guanosine-3',5'-cyclic monophosphate, Rp-isomer $(Rp-pCPT-cGMP,\;100\;{\mu}M),$ a potent inhibitor of PKG or protein phosphatase 2A (PP2A, 1 U/ml). In the presence of okadaic acid (OA, 5 nM), PP2A had no effect on the channel activity. The $K_{Ca}$ channel activity spontaneously decayed to the control level upon washout of ATP, cGMP and PKG, and this was prevented by OA (5 nM) in the medium. These results suggest that the PKG-mediated phosphorylations of $K_{Ca}$ channels, or some associated proteins in the membrane patch increase the activity of the $K_{Ca}$ channel, and the activation may be associated with the vasodilating action.

• The Korean Journal of Physiology and Pharmacology
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• 제24권4호
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• pp.329-338
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• 2020

Rhinorrhea in allergic rhinitis (AR) is characterized by the secretion of electrolytes in the nasal discharge. The secretion of Cl^- and HCO₃^- is mainly regulated by cystic fibrosis transmembrane conductance regulator (CFTR) or via the calcium-activated Cl^- channel anoctamin-1 (ANO1) in nasal gland serous cells. Interleukin-4 (IL-4), which is crucial in the development of allergic inflammation, increases the expression and activity of ANO1 by stimulating histamine receptors. In this study, we investigated ANO1 as a potential therapeutic target for rhinorrhea in AR using an ANO1 inhibitor derived from a natural herb. Ethanolic extracts (30%) of Spirodela polyrhiza (SP_EtOH) and its five major flavonoids constituents were prepared. To elucidate whether the activity of human ANO1 (hANO1) was modulated by SP_EtOH and its chemical constituents, a patch clamp experiment was performed in hANO1-HEK293T cells. Luteolin, one of the major chemical constituents in SP_EtOH, significantly inhibited hANO1 activity in hANO1-HEK293T cells. Further, SP_EtOH and luteolin specifically inhibited the calcium-activated chloride current, but not CFTR current in human airway epithelial Calu-3 cells. Calu-3 cells were cultured to confluency on transwell inserts in the presence of IL-4 to measure the electrolyte transport by Ussing chamber. Luteolin also significantly inhibited the ATP-induced increase in electrolyte transport, which was increased in IL-4 sensitized Calu-3 cells. Our findings indicate that SP_EtOH and luteolin may be suitable candidates for the prevention and treatment of allergic rhinitis. SP_EtOH^- and luteolin-mediated ANO1 regulation provides a basis for the development of novel approaches for the treatment of allergic rhinitis-induced rhinorrhea.

• 대한수의학회지
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• 제34권1호
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• pp.25-36
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• 1994

The inward tail current after a short depolarizing pulse has been known as Na-Ca exchange current activated by intracellular calcium which forms late plateau of the action potential in rabbit atrial myocytes. Chloride conductance which is also dependent upon calcium concentration has been reported as a possible tail current in many other excitable tissues. Thus, in order to investigate the exsitance of the calcium activated chloride current and its contribution to tail current, whole cell voltage clamp measurement has been made in single atrial cells of the rabbit. The current was recorded during repolarization following a brief 2 ms depolarizing pulse to +40mV from a holding potential of -70mV. When voltage-sensitive transient outward current was blocked by 2 mM 4-aminopyridine or replacement potassium with cesium, the tail current were abolished by ryanodine$(1{\mu}M)$ or diltiazem$(10{\mu}M)$ and turned out to be calcium dependent. The magnitudes of the tail currents were increased when intracellular chloride concentration was increased to 131 mM from 21 mM. The current was decreased by extracellular sodium reduction when intracellular chloride concentration was low(21 mM), but it was little affected by extracellular sodium reduction when intracellual chloride concentration was high(131 mM). The current-voltage relationship of the difference current before and after extracellular sodium reduction, shows an exponential voltage dependence with the largest magnitude of the current occurring at negative potentials, with is similar to current-voltage relationship at negative potentials, which is similar to current-voltage relationship of Na-Ca exchange current. The current was also decreased by $10{\mu}M$ niflumic acid and 1 mM bumetanide, which is well known anion channel blockers. The reversal potentials shifted according to changes in chloride concentration. The current-voltage relationships of the niflumic acid-sensitive currents in high and low concentration of chloride were well fitted to those predicted as chloride current. From the above results, it is concluded that calcium activated chloride component exists in the tail current with Na-Ca exchange current and it shows the reversal of tail current. Therefore it is thought that in the physiologic condition it leads to rapid end of action potential which inhibits calcium influx and it contributes to maintain the low intracellular calcium concentration with Na-Ca exchange mechanism.

• 한국진공학회:학술대회논문집
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• 한국진공학회 2011년도 제40회 동계학술대회 초록집
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• pp.27-27
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• 2011

For the design of real applicable molecular devices, current-voltage properties through molecular nanostructures such as metal-molecule-metal junctions (molecular junctions) have been studied extensively. In thiolate monolayers on the gold electrode, the chemical bonding of sulfur to gold and the van der Waals interactions between the alkyl chains of neighboring molecules are important factors in the formation of well-defined monolayers and in the control of the electron transport rate. Charge transport through the molecular junctions depends significantly on the energy levels of molecules relative to the Fermi levels of the contacts and the electronic structure of the molecule. It is important to understand the interfacial electron transport in accordance with the increased film thickness of alkyl chains that are known as an insulating layer, but are required for molecular device fabrication. Thiol-tethered RuII terpyridine complexes were synthesized for a voltage-driven molecular switch and used to understand the switch-on mechanism of the molecular switches of single metal complexes in the solid-state molecular junction in a vacuum. Electrochemical voltammetry and current-voltage (I-V) characteristics are measured to elucidate electron transport processes in the bistable conducting states of single molecular junctions of a molecular switch, Ru(II) terpyridine complexes. (1) On the basis of the Ru-centered electrochemical reaction data, the electron transport rate increases in the mixed self-assembled monolayer (SAM) of Ru(II) terpyridine complexes, indicating strong electronic coupling between the redox center and the substrate, along the molecules. (2) In a low-conducting state before switch-on, I-V characteristics are fitted to a direct tunneling model, and the estimated tunneling decay constant across the Ru(II) terpyridine complex is found to be smaller than that of alkanethiol. (3) The threshold voltages for the switch-on from low- to high-conducting states are identical, corresponding to the electron affinity of the molecules. (4) A high-conducting state after switch-on remains in the reverse voltage sweep, and a linear relationship of the current to the voltage is obtained. These results reveal electron transport paths via the redox centers of the Ru(II) terpyridine complexes, a molecular switch.

• 지질공학
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• 제25권3호
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• pp.413-420
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• 2015

안성천에 위치한 공도 수위표 상류 구간에 대해 하상퇴적층을 통과하는 하천수-지하수 상호교환량과 수리경사를 측정하여 하상퇴적층의 수리전도도를 산정하였다. 하천수-지하수 상호유동량은 자체 제작한 시피지 미터를 이용하여 측정하였고, 수리경사는 장심도와 단심도 두 가지 종류의 관입형 피조미터를 하상퇴적층에 설치하여 피조미터내 수두 차이와 관입깊이 차이를 측정하여 산정하였다. 하천수-지하수 교환량은 -1.55 × 10^-6~1.77 × 10^-5 m/s, 연직방향 수리경사는 -0.122에서 0.030로 측정되었다. 교환량과 수리경사로 부터 연직방향 수리전도도를 산정한 결과 1.77 × 10^-5~1.97 × 10^-3 m/s로 지점별로 큰 차이를 나타내었다. 측정 결과는 Calver (2001)가 제시한 하상퇴적물 수리전도도의 일반적인 범위내에서 분포하였으며, 모래, 자갈층이 잘 발달되어 있어 기존의 국내 측정 결과 사례에 비해 큰 값을 보이는 것으로 분석되었다. 본 논문에서 적용한 고강도 스테인레스 재질의 피조미터와 시피지미터를 이용하여 하상퇴적층의 수리전도도를 측정하는 방법은 내구성과 휴대성이 우수하여 실무에서 매우 유용하게 활용될 수 있을 것으로 기대된다.

• 감성과학
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• 제5권2호
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• pp.29-34
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• 2002

이전 연구에서 가상현실에 몰입하는 동안 넓은 시야(field of view : 150˚)와 빠른 운행속도(70km/sec)가 사이버 멀미를 심화시킨다는 결과를 얻었다. 피험자의 90%가 좁은 시야(50˚)와 느린 운행속도(30km/sec)에서 사이버멀미 증상이 적었다. 본 실험에서는 피험자가 생리적인 동요를 경험할 때마다 바이오피드백 방법을 사용하여 사이버 멀미 감소 가상환경(cybersickness alleviating virtual environment ; CAVE)을 제시한 후, 그 효과를 관찰하였다. 피부전도도, 말초체온, 말초 혈류량, 심박률, 눈 깜박임, 뇌전위의 변수들을 입력하는 인공 신경망으로 구성된 실시간 멀미 탐지 시스템과 CAVE-제시 피드백 시스템을 구축하였다 이 시스템들은 생리적 측정치들이 사이버 멀미의 출현을 신호할 때마다 피드백 출력으로 좁은 화면과 감소된 운행속도를 일시적으로 제공하였다. 36명의 피험자를 대상으로 SSQ(simulator sickness questionnaire)와 자기보고를 이용하여 사이버 멀미의 빈도와 심각도를 조사하였다. 모든 피험자는 한달 간격으로 CAW 조건과 non-CAYE 조건에서 두 번 가상현실을 경험하였다. 사이버멀미의 빈도와 심각도는 non-CAVE 조건보다 CAVE 조건에서 유의미하게 감소하였다. 즉, 전기 생리학적 특징들에 기반한 인공 신경망에 의해 제공된 좁은 시야와 느린 운행의 가상환경은 사이버 멀미 증상들을 감소시켰다 이러한 결과들은 생체신호 피드백 시스템을 이용하여 인간 친화적 가상환경을 구축할 수 있는 가능성을 보인 것이다.

• 한국농림기상학회지
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• 제21권4호
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• pp.261-268
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• 2019

한발 발생이 갈수록 증가하고 있는 가운데, 작물의 생리생화학적 기작과 광합성능을 효율적으로 진단 및 평가 할 수 있는 방법을 찾는 것은 매우 중요한 과제이다. 본 연구를 통해 광화학 반사 지수인 PRI가 콩의 한발 스트레스에 대하여 고온과 고CO₂ 등 미래 기후 변화 환경에서도 유의미한 생육 지표로 활용될 수 있음을 확인하였다. PRI와 대표적 한발 생육지표들(군락 광이용효율, 건물중 변동율, 엽록소 형광, 기공전도도)과의 밀접한 관계를 통해 한발 스트레스 하에서 콩의 광합성 활성과 생장량을 원격으로 모니터링할수 있는 진단 모델 개발의 가능성을 볼 수 있었다. 다만 향후 고온, 고CO₂ 등 기후변화 조건에서 PRI의 성능에 대한 지속적인 평가와 함께 민감도 향상을 위한 광학지표 개발 및 모델 개선 연구가 선행되어야 할 것으로 보인다.

• 환경생물
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• 제20권4호
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• pp.303-308
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• 2002

3-weeks old Commelina was transferred to and grown in Hoagland solution ($\pm$ 100 $\mu{M}$ $Cd^{2+}$, 100 $\mu{M}$ $Cd^{2+}$ ＋ 100$\mu{M}$ $Ca^{2+}$, 100 $\mu{M}$ $Cd^{2+}$ ＋ 200 $\mu{M}$ EGTA) for two weeks and then a number of physiological activities was investigated. $Cd^{2+}$ reduced total chlorophyll content up to 29% at a week and 75% at two weeks. In the treatment of $Cd^{2+}$ ＋ $Ca^{2+}$, the total chlorophyll content was reduced to 29% at a week and 80% at two weeks. $Cd^{2+}$ reduced 24% of Fv/Fm after two weeks. In case of $Cd^{2+}$ ＋ $Ca^{2+}$, Fv/Fm was reduced 55% at a week, but after two weeks, the plants were almost dead and Fv/Fm could not be measured. When EGTA was treated with $Cd^{2+}$, the value of Fv/Fm was not affected. There were no differences of water potential between the control and the treatment of $Cd^{2+}$+EGTA toy a week, but in other treatments. water potential was reduced. $Cd^{2+}$ reduced about 21% of water potential and $Cd^{2+}$ ＋ $Ca^{2+}$ reduced 43% of water potential after two weeks. $Cd^{2+}$ inhibited 21% of photosynthetic activity at a week and 32% at two weeks. In case of photosynthetic activity, $Cd^{2+}$ ＋ $Ca^{2+}$ inhibited 58% at a week and 73% at two weeks. $Cd^{2+}$+EGTA inhibited 15% of photosynthetic activity at a week and 21% at two weeks. Similar results were found in stomatal conductance. From the above results, it was observed that the treatment of $Ca^{2+}$ with $Cd^{2+}$ induced more reduction of a series of physiological responses than those of the treatment of $Cd^{2+}$ alone. Therefore, it could be concluded that $Ca^{2+}$ did not reduce the toxicity of $Cd^{2+}$, but enhanced $Cd^{2+}$ -induced physiological toxicities, but EGTA induced an decrease of $Cd^{2+}$ -induced physiological toxicities.

• 한국농림기상학회지
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• 제7권4호
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• pp.296-302
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• 2005

건조 스트레스에 의한 광합성 기구의 피해 양상을 파악하기 위하여 사시나무를 대상으로 광합성 특성의 변화를 조사하였다. 사시나무의 뿌리를 채취하여 근 맹아를 유도하고 각 개체를 증식한 후 실험에 사용하였다. 건조처리는 PEG농도를 무처리구인 대조구와 $0\%,\;2\%,\;5\%,\;and\;10\%$ 처리구로 구분하여, 주1회 300ml씩 관수하여 4주 후 광합성 특성과 광화학계 및 탄소고정계를 분석하였다 수분스트레스의 강도가 증가함에 따라 사시나무의 광합성속도, 기공전도도 및 증산속도는 모두 감소하는 경향을 보였다. 광화학계에서 PEG 처리농도의 증가는 PSII의 양자수율을 감소시켰으며, 이러한 전자에너지 전달의 감소는 순양자수율도 감소시켰다. 한편 호흡속도는 PEG 처리 농도가 증가함에 따라 감소하였으나, 탄소고정계의 광호흡속도는 반대로 증가하였다. 결론적으로 사시나무는 건조스트레스를 받으면 광합성관련 기구가 매우 민감하게 반응하며, 그들의 민감성은 수분스트레스의 강도에 좌우된다. 한편 사시나무는 수분이용효율의 증가와 같은 수분스트레스에 대한 내성 조절 능력을 보여준다.

• 생명과학회지
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• 제27권11호
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• pp.1349-1356
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• 2017

$K^+$ 통로 개방제들은 심근, 뇌, 골격근 등에서 세포막 혹은 미토콘드리아 내막에 존재하는 큰 전도성의 $Ca^{2+}$-의 존성 $K^+$ (BK) 통로 및 ATP-조절성 $K^+$ 통로(ATP-sensitive $K^+$ channels, $K_{ATP}$)에 작용하여 허혈성 혹은 산화성 세포 손상을 완화하는 효과가 있는 것으로 보고되어 있다. 본 연구에서는 망막 색소 상피세포주인 ARPE-19 세포를 실험 모델로 하여 큰 전도성의 BK 통로 개방제인 NS 1619가 유사한 보호 효과를 나타낼 수 있는지, 또한 그 작용기전이 무엇인지를 확인하고자 하였다. AREE-19 세포를 여러 형태의 산화 스트레스에 노출시켜 세포 손상을 유발하고 그 손상의 정도 및 이에 미치는 NS 1619의 효과를 trypan blue 배출능, Tunel 염색 분석을 통하여 측정하였다. NS 1619는 여러 형태의 산화 스트레스에 의한 괴사성 및 apoptosis에 의한 세포 손상을 효과적으로 방지하였으며 그 보호 효과는 BK 통로 봉쇄제인 paxilline 의해 차단되었다. NS 1619는 $H_2O_2$에 의한 세포내 ATP 고갈을 현저히 완화시켰으며, 또한 MTT 환원능으로 측정한 미토콘드리아의 기능을 보호하는 효과를 보였다. 유세포형광 분석법을 이용한 실험에서 NS 1619는 $H_2O_2$에 의한 미토콘드리아 막전압의 소실을 유의하게 방지하였다. 이상의 결과들을 종합하면 NS 1619는 망막 색소 상피세포에서 산화성 세포 손상을 방지하는 효과를 나타내며 그 기전에 미토콘드리아 기능에 대한 보호 작용이 연관되어 있는 것으로 사료된다.