• 한국자기공명학회논문지
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• 제23권3호
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• pp.81-86
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• 2019

Overhauser dynamic nuclear polarization (O-DNP) has been an efficient method to boost the thermal nuclear polarization in liquids at room temperature. However, O-DNP for a benchtop NMR using a permanent magnet has remained unexplored yet. In this work, we report the development of an O-DNP system adopting a permanent magnet of 1.6 T. Q-band (~43 GHz) high-power amplifier produced 6 W microwave for saturation. Instead of resonator, we used an open-type antenna for the microwave irradiation. For several representative small molecules, we measured the concentration and frequency dependences of the enhancement factor. This work paves the way for the development of a benchtop DNP-NMR system overcoming its disadvantage of low quality signal when using a permanent magnet.

• Journal of Microbiology and Biotechnology
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• 제33권7호
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• pp.857-863
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• 2023

Pathogenic bacteria that colonize the human intestinal tract have evolved strategies to overcome acidic conditions when they pass through the gastrointestinal tract. Amino acid-mediated acid resistance systems are effective survival strategies in a stomach that is full of amino acid substrate. The amino acid antiporter, amino acid decarboxylase, and ClC chloride antiporter are all engaged in these systems, and each one plays a role in protecting against or adapting to the acidic environment. The ClC chloride antiporter, a member of the ClC channel family, eliminates negatively charged intracellular chloride ions to avoid inner membrane hyperpolarization as an electrical shunt of the acid resistance system. In this review, we will discuss the structure and function of the prokaryotic ClC chloride antiporter of amino acid-mediated acid resistance system.

• 한국수정란이식학회지
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• 제21권1호
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• pp.35-43
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• 2006

이온 통로 및 이온 농도의 변화는 수정 현상을 포함한 다양한 세포 기능에 중요한 역할을 한다. 그러나 이러한 이온의 변화가 포유동물 배의 발달과정에 어떻게 관여하는지에 대해서는 알려진 바가 적다. 본 연구에서는 생쥐난자가 수정 이후 배 발달 과정을 거치는 동안 나타나는 칼슘과 포타슘 이온의 변화를 전기생리학적 실험 기법과 공초점 현미경을 이용하여 조사하였다. 수정 시에 나타나는 일시적인 세포내 칼슘 농도 변화는 활성 전류(수정 전류)와 함께 동반되었다. 그러나 수정과 같은 극적인 현상이나 자극이 없는 시기에는 세포내 칼슘 농도가 배 발달 시기와 상관없이 일정한 수준으로 유지되었다. 이것은 세포내외의 칼슘 농도의 보상현상으로도 설명할 수 있을 것이다. 배 발달이 진행됨에 따라 난관액의 포타슘 농도는 계속 증가하여 8세포기 배에서는 난자보다 26% 증가하였다. 상실배, 포배기에서는 포타슘 농도가 감소하였다. 배 발달이 진행됨에 따라 주로 포타슘 이온에 의해 조절되는 막 전압은 탈분극되고, 칼슘 이온의 세포 안으로의 유입은 점점 감소하였다. 생쥐 난자에 5 mM의 칼슘을 처리하였을 때 막 전압은 일시적인 과분극 현상을 보이다가 회복되었다. 칼슘 유입에 따른 막 전압 변화에 관여하는 포타슘 통로를 확인하기 위하여 포타슘 통로 차단제를 전 처리한 후 칼슘을 처리한 결과, 칼슘만을 단독으로 처리한 결과와 유의한 차이를 보이지 않았다. 막 전압의 과분극 현상은 잘 알려진 포타슘 통로 차단제인 TEA에 억제되지 않았다. 그리고 small conductance $Ca^{2+}$-activated 포타슘 통로 차단제 인 apamin에 의해서도 억제되지 않았다. 따라서 생쥐 난자에서 과분극을 유발시키는 포타슘 통로는 TEA와 apamin에 억제되지 않는 다른 포타슘 통로로 생각된다. 이상의 결과로부터 배 발달 동안 변화되는 칼슘과 포타슘 이온은 수정 및 초기 배 발달에 중요한 인자로써 작용할 것으로 생각되며, two-pore domain 포타슘 통로가 난자의 막 전압 조절에 관여할 가능성을 제시한다.

• The Korean Journal of Physiology and Pharmacology
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• 제19권5호
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• pp.435-440
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• 2015

This study aimed to investigate the effect of pituitary adenylate cyclase-activating peptide (PACAP) on the pacemaker activity of interstitial cells of Cajal (ICC) in mouse colon and to identify the underlying mechanisms of PACAP action. Spontaneous pacemaker activity of colonic ICC and the effects of PACAP were studied using electrophysiological recordings. Exogenously applied PACAP induced hyperpolarization of the cell membrane and inhibited pacemaker frequency in a dose-dependent manner (from 0.1 nM to 100 nM). To investigate cyclic AMP (cAMP) involvement in the effects of PACAP on ICC, SQ-22536 (an inhibitor of adenylate cyclase) and cell-permeable 8-bromo-cAMP were used. SQ-22536 decreased the frequency of pacemaker potentials, and cell-permeable 8-bromo-cAMP increased the frequency of pacemaker potentials. The effects of SQ-22536 on pacemaker potential frequency and membrane hyperpolarization were rescued by co-treatment with glibenclamide (an ATP-sensitive $K^+$ channel blocker). However, neither $N^G$-nitro-L-arginine methyl ester (L-NAME, a competitive inhibitor of NO synthase) nor 1H-[1,2,4]oxadiazolo[4,3-${\alpha}$]quinoxalin-1-one (ODQ, an inhibitor of guanylate cyclase) had any effect on PACAP-induced activity. In conclusion, this study describes the effects of PACAP on ICC in the mouse colon. PACAP inhibited the pacemaker activity of ICC by acting through ATP-sensitive $K^+$ channels. These results provide evidence of a physiological role for PACAP in regulating gastrointestinal (GI) motility through the modulation of ICC activity.

• 한국생물물리학회:학술대회논문집
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• 한국생물물리학회 2003년도 정기총회 및 학술발표회
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• pp.39-39
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• 2003

In our previous study, WEHI-231, an immature B cell line, showed intractable increase in [C $a^{2+}$]$_{c}$ after the B-cell receptor (BCR) ligation and treatment with 2-aminoethoxydiphenylborate (2-APB), which was never observed in Bal-17, a mature B cell line (Nam et al., 2003, FEBS Lett). In this study, a whole cell voltage clamp study revealed a specific expression of a novel type of $K^{+}$ current, namely voltage-independent background-type $K^{+}$ channels (IK-bg), in WEHI-231 cells. IK-bg was dramatically increase by the application of 2-APB (50 $\square$M), which induced severe hyperpolarization of WEHI-231 from -45 ㎷ to -90 ㎷, When dialyzed with $Mg^{2+}$ and ATP-free pipette solution, a spontaneous development of IK-bg and membrane hyperpolarization were observed. IK-bg was insensitive to classical $K^{+}$ channel blockers (TEA, glibenclamide, $Ba^{2+}$(1 mM)), whereas blocked by quinine and quinidine in a voltage-dependent manner ($IC_{50}$/=6~9 $\square$M at +60㎷). Phorbol myrstate, a PKC activator, decreased the amplitude of IK-bg. Extracellular acidification (pH 6.5) slightly inhibited IK-bg. Arachidonic acid, riluzole, or hyposmotic stress could not affect the IK-bg after the full development by the intracellular dialysis with Mg-ATP-free solution. In a cell-attached mode of single channel recording from WEHI231, we found two types of voltage-independent $K^{+}$ channels with unitary conductance of 300 pS and 120 pS, respectively. Both channels showed very short mean open times and their open probabilities were increase by the application of 2-APB. In Bal-17 cells, no such $K^{+}$ current was observed in 50 cells tested. In summary, WEHI-231 immature B cells express background $K^{+}$ channels. The pharmacological properties and the large unitary conductance suggest that novel types of two-pore domain $K^{+}$ channels (2-P-K channels) might be expressed in WEHI-231, which may provide an intriguing targets of signal transduction in the immature B lymphocytes.e B lymphocytes.

• Bulletin of the Korean Chemical Society
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• 제33권2호
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• pp.511-514
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• 2012

A continuous flow hyperpolarized (HP) $^{129}Xe$ NMR spectroscopy was employed for the first time to investigate $TiO_2$ nanotubes (Ti-NTs) synthesized from commercial nanoparticles with different reaction times. A single peak attributing to channels for Ti-NTs was observed for variable temperature HP $^{129}Xe$ NMR spectra. It was also noted that there is alteration in value for heat of adsorption, ${\Delta}H$ from $12.6{\pm}1.3$ to $16.4{\pm}0.4kJ/mol$ and variation in chemical shift of the xenon adsorbed in channels, ${\delta}_s$ from $120{\pm}2\sim135{\pm}9ppm$ which were closely correlated to channel length and it was shown that P25-24 Ti-NTs with longest channel is most favorite Ti-NTs for Xe adsorption.

• 대한약리학회지
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• 제30권2호
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• pp.181-190
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• 1994

In rat atrium the characteristics of purinergic receptors were investigated by observing the effects of some purinergic receptor agonists and antagonists on action potential and contractile force. The statistically significant effects of $ATP(10^{-6}{\sim}10^{-3}M)$ and adenosine $(10^{-6}{\sim}10^{-3}M)$ on normal action potential characteristics were a dose-dependent shortening of action potential duration $(APD_{90})$ by both agents and hyperpolarization by $ATP(10^{-4},10^{-3}M)$. $CAP(10^{-8}{\sim}10^{-4}M)$, an $A_1$ adenosine receptor agonist, shortened $(APD_{90})$ markedly in a dose-dependent manner and these effects were almost abolished by $DPCPX\;(10^{-6}\;M), an $A_1$, adenosine receptor antagonist, but not affected by $DMPX(2{\times}10^{-6}\;M)$, an $A_2$ adenosine receptor agonist. On the other hand, CGS $21680(10^{-7}{\sim}10^{-4}M)$, an $A_2$ adenosine receptor agonist, elicited a slight shortening of $(APD_{90})$ and these effects were inhibited by DPCPX but persisted in the presence of DPMX. Adenosine $(10^{-6}{\sim}10{\-4}\;M)$ decreased the basal contraction of atrial muscle in a dose-dependent manner and these effects were not inhibited by DMPX but by DPCPX. These results suggests that purinergic receptor agonists depress the cardiac activity by a short ening of action potential duration and this effect is mostly mediated by $A_1$ adenosine receptors in rat atrium.

• 대한의생명과학회지
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• 제17권3호
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• pp.177-184
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• 2011

Potassium cyanate (KOCN) is an inorganic compound and induces the carbamylation of proteins with cytotoxic effects on human cells. Although there is a potential cytotoxic molecule, the role of KOCN on the apoptosis of cancer cell is not well understood. The present study investigated the effects of KOCN on the human colorectal cancer cell line, HCT 116 cells. To understand the anti-cancer effect of KOCN on HCT 116 cells, we examined alteration of apoptosis, the intracellular $Ca^{2+}$ concentration, the intracellular signaling pathway and generation of reactive oxygen species (ROS) in these cells treated with KOCN. The apoptosis of HCT 116 cells was induced by KOCN in a dose-dependent manner at 24 hours and 48 hours, respectively. The apoptosis was processed via the cleavage of poly ADP-ribose polymerase (PARP) and activation of caspase 3 in HCT 116 cells. KOCN induced the elevation of intracellular $Ca^{2+}$ concentration and changed the expressions of Bcl-2 family proteins. The pro-apoptotic Bax was continuously up-regulated, and the anti-apoptotic Bcl-2 was down-regulated by KOCN. KOCN also induced the hyperpolarization of mitochondria and the generation of ROS in HCT 116 cells. Taken together, these results indicate that KOCN induces the apoptosis of HCT 116 cells by disruption of $Ca^{2+}$ homeostasis and via mitochondrial pathway. This study provides the compound that may be used as a potent agent for the treatment of colorectal cancer.

• BMB Reports
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• 제45권11호
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• pp.635-640
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• 2012

To understand the effects of HCN as potential mediators in the pathogenesis of epilepsy that evoke long-term impaired excitability; the present study was designed to elucidate whether the alterations of HCN expression induced by status epilepticus (SE) is responsible for epileptogenesis. Although HCN1 immunoreactivity was observed in the hippocampus, its immunoreactivities were enhanced at 12 hrs following SE. Although, HCN1 immunoreactivities were reduced in all the hippocampi at 2 weeks, a re-increase in the expression at 2-3 months following SE was observed. In contrast to HCN1, HCN 4 expressions were un-changed, although HCN2 immunoreactive neurons exhibited some changes following SE. Taken together, our findings suggest that altered expressions of HCN1 following SE may be mainly involved in the imbalances of neurotransmissions to hippocampal circuits; thus, it is proposed that HCN1 may play an important role in the epileptogenic period as a compensatory response.

• The Korean Journal of Physiology and Pharmacology
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• 제16권1호
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• pp.71-77
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• 2012

Mitochondrial dynamics and distribution is critical for their role in bioenergetics and cell survival. We investigated the consequence of altered fission/fusion on mitochondrial function and motility in INS-1E rat clonal ${\beta}$-cells. Adenoviruses were used to induce doxycycline-dependent expression of wild type (WT-Mfn1) or a dominant negative mitofusin 1 mutant (DN-Mfn1). Mitochondrial morphology and motility were analyzed by monitoring mitochondrially-targeted red fluorescent protein. Adenovirus-driven overexpression of WT-Mfn1 elicited severe aggregation of mitochondria, preventing them from reaching peripheral near plasma membrane areas of the cell. Overexpression of DN-Mfn1 resulted in fragmented mitochondria with widespread cytosolic distribution. WT-Mfn1 overexpression impaired mitochondrial function as glucose- and oligomycin-induced mitochondrial hyperpolarization were markedly reduced. Viability of the INS-1E cells, however, was not affected. Mitochondrial motility was significantly reduced in WT-Mfn1 overexpressing cells. Conversely, fragmented mitochondria in DN-Mfn1 overexpressing cells showed more vigorous movement than mitochondria in control cells. Movement of these mitochondria was also less microtubule-dependent. These results suggest that Mfn1-induced hyperfusion leads to mitochondrial dysfunction and hypomotility, which may explain impaired metabolism-secretion coupling in insulin-releasing cells overexpressing Mfn1.