• Title/Summary/Keyword: resting cells

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Effects of Allicin on Cytokine Production Genes of Human Peripheral Blood Mononuclear Cells (마늘의 Allicin이 사람 단핵세포의 사이토카인 생산 유전자의 발현에 미치는 영향)

  • 박란숙
    • The Korean Journal of Food And Nutrition
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    • v.15 no.3
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    • pp.191-196
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    • 2002
  • The effect of allicin, the major component of garlic (Allium sativum), on the gene expression profiles of peripheral blood mononuclear cells from healthy donors was analyzed. DNA microarray which can detect expression signal of 862 genes revealed that allicin induced the expression of cytokine, chemokine, and immune-related genes in peripheral blood mononuclear cells. In contrast, allicin repressed the expression of adaptive immune-related genes, which are expressed in T helper 1 Iymphocytes. Simultaneous inhibitory and stimulatory effects of allicin were found on inflammatory cells. It is likely that allicin down-regulated the expression of specific genes that were previously up-regulated in resting cells, suggesting a new mechanism by which they exert positive and negative effect. Considering the broad and renewed interest in allicin, the profiles we describe here will be useful in designing more specific and efficient treatment strategies.

Regional Differences in Voltage-tension Relationship of Gastric Smooth Muscles in Guinea-pig (위 평활근의 부위별 전압-장력 관계에 관한 연구)

  • Kim, Ki-Whan;Lee, Sang-Jin;Suh, Suk-Hyo
    • The Korean Journal of Physiology
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    • v.23 no.2
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    • pp.263-275
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    • 1989
  • Mechanical contractions and electrical activities of the fundic longitudinal and antral circular muscle fibers were investigated in order to elucidate topical differences of gastric motility. K-induced contracture was produced by exposure of muscle strips to high K Tyrode solution. Membrane potential and mechanical contraction were simultaneously recorded by conventional glass microelectrode method and single sucrose-gap technique. All experiments were performed in tris-buffered Tyrode solution which was aerated with $100%\;O_2\;and\;kept\;35^{\circ}C$. The results obtained were as follows: 1) The resting membrane potential of circular muscle cells in the antral region was about 10 mV more negative than that in the fundic region. 2) The membrane potentials decreased almost linearly as the extracellular KCI concentration was increased both in antral circular muscle cells and in fundic longitudinal muscle cells. 3) The thresholdal K concentration of K-contracture was 15 mM (membrane potential, -48 mV) for the antral circular muscle strip and 20 mM for the fundic longitudinal muscle cells. 4) The ratio of membrane permeability coefficient for $Na^+\;and\;K^+,\;P_{Na}/P_K\;({\alpha})$ was 0.065 for antral circular muscle cells and was 0.108 for fundic longitudinal muscle cells. 5) K-contracture of antral and fundic smooth muscle strips showed the contracture composed of phasic and tonic components. The amplitude of the phasic component increased sigmoidally in a dose-dependent manner, whereas that of the tonic component was maximal at a concentration of 40 mM KCI and at the concentrations above or below 40 mM KCI the amplitude was reduced. 6) The inverse relationship between the amplitude of tonic component and extracellular KCI concentration in the range of 40 to 150 mM KCI was more prominent in the antral circular muscle strip than in the fundic longitudinal muscle strip, where the amplitude of the tonic component decreased less steeply and was maintained higher at the same high K concentrations. 7) The tonic component was totally dependent on the external $Ca^{2+}$ and completely abolished by verapamil, while tile phasic component was far less dependent on the external $Ca^{2+}$ and partially suppressed by verapamil. From the above results, the following conclusions could be made. 1) The phasic component of K-contracture is produced both by intracellular $Ca^{2+}$ mobilization and by $Ca^{2+}$-influx from outside, while the tonic component is generated and maintained by the $Ca^{2+}-influx$ through the potential-dependent $Ca^{2+}$ channel. 2) The mechanism of reducing the free $Ca^{2+}$ concentration in the myoplasm seems to be more developed in the antral circular muscle than in the fundic longitudinal muscle. 3) The lower resting membrane potential of the fundic longitudinal muscle cell reflects a relatively high $P_{Na}/P_K$ ratio of about 0.108.

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Bone Marrow-derived Side Population Cells are Capable of Functional Cardiomyogenic Differentiation

  • Yoon, Jihyun;Choi, Seung-Cheol;Park, Chi-Yeon;Choi, Ji-Hyun;Kim, Yang-In;Shim, Wan-Joo;Lim, Do-Sun
    • Molecules and Cells
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    • v.25 no.2
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    • pp.216-223
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    • 2008
  • It has been reported that bone marrow (BM)-side population (SP) cells, with hematopoietic stem cell activity, can transdifferentiate into cardiomyocytes and contribute to myocardial repair. However, this has been questioned by recent studies showing that hematopoietic stem cells (HSCs) adopt a hematopoietic cell lineage in the ischemic myocardium. The present study was designed to investigate whether BM-SP cells can in fact transdifferentiate into functional cardiomyocytes. Phenotypically, BM-SP cells were $19.59%{\pm}9.00\;CD14^+$, $8.22%{\pm}2.72\;CD34^+$, $92.93%{\pm}2.68\;CD44^+$, $91.86%{\pm}4.07\;CD45^+$, $28.48%{\pm}2.24\;c-kit^+$, $71.09%{\pm}3.67\;Sca-1^+$. Expression of endothelial cell markers (CD31, Flk-1, Tie-2 and VEGF-A) was higher in BM-SP cells than whole BM cells. After five days of co-culture with neonatal cardiomyocytes, $7.2%{\pm}1.2$ of the BM-SP cells expressed sarcomeric ${\alpha}$-actinin as measured by flow cytometry. Moreover, BM-SP cells co-cultured on neonatal cardiomyocytes fixed to inhibit cell fusion also expressed sarcomeric ${\alpha}$-actinin. The co-cultured BM-SP cells showed neonatal cardiomyocyte-like action potentials of relatively long duration and shallow resting membrane potential. They also generated calcium transients with amplitude and duration similar to those of neonatal cardiomyocytes. These results show that BM-SP cells are capable of functional cardiomyogenic differentiation when co-cultured with neonatal cardiomyocytes.

The Role of Intracellular $Mg^{2+}$ in Regulation of $Ca^{2+}-activated$ $K^+$ Channel in Pulmonary Arterial Smooth Muscle Cells of the Rabbit

  • Lee, Suk-Ho;Park, Myoung-Kyu
    • The Korean Journal of Physiology and Pharmacology
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    • v.2 no.5
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    • pp.611-616
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    • 1998
  • Although the $Ca^{2+}-activated\;K^+\;(I_{K,Ca})$ channel is known to play an important role in the maintenance of resting membrane potential, the regulation of the channel in physiological condition is not completely understood in vascular myocytes. In this study, we investigated the role of cytoplasmic $Mg^{2+}$ on the regulation of $I_{K,Ca}$ channel in pulmonary arterial myocytes of the rabbit using the inside-out patch clamp technique. $Mg^{2+}$ increased open probability (Po), but decreased the magnitude of single channel current. $Mg^{2+}-induced$ block of unitary current showed strong voltage dependence but increase of Po by $Mg^{2+}$ was not dependent on the membrane potential. The apparent effect of $Mg^{2+}$ might, thus, depend on the proportion between opposite effects on the Po and on the conductance of $I_{K,Ca}$ channel. In low concentration of cytoplasmic $Ca^{2+},\;Mg^{2+}$ increased $I_{K,Ca}$ by mainly enhancement of Po. However, at very high concentration of cytoplasmic $Ca^{2+},$ such as pCa 5.5, $Mg^{2+}$ decreased $I_{K,Ca}$ through the inhibition of unitary current. Moreover, $Mg^{2+}$ could activate the channel even in the absence of $Ca^{2+}.\;Mg^{2+}$ might, therefore, partly contribute to the opening of $I_{K,Ca}$ channel in resting membrane potential. This phenomenon might explain why $I_{K,Ca}$ contributes to the resting membrane potential where membrane potential and concentration of free $Ca^{2+}$ are very low.

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Induction of PCB degradative pathway by plant terpenoids as growth substrates or inducers

  • Jeong, Gyeong-Ja;Kim, Eung-Bin;So, Jae-Seong;Go, Seong-Cheol
    • 한국생물공학회:학술대회논문집
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    • 2000.11a
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    • pp.489-492
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    • 2000
  • The eventual goal of this study is to elucidate roles of plant terpenoids (e.g., cymene, limonene and others) as natural substrates in the cometabolic biodegradation of PCBs and to develop an effective PCB bioremediation technology. The aim of this study was to examine how plant terpenoids, as natural substrates or inducers would affect the biodegradation of PCB congeners. Various PCB degraders that could grow on biphenyl and several terpenoids were tested for their PCB degradation capabilities. The PCB congener degradation activities were first monitored through resting cell assay technique that could detect degradation products of the substrate. The congener removal was also confirmed by concommitant GC analysis. The PCB degraders, Pseudononas sp. P166 and Caynebacterium sp. T104 were found to grow on both biphenyl and terpenoids ((S)-(-) limonene, p-cymene and ${\alpha}-terpinene$) whereas Arthrobacter B1B could not grow on the terpenoids as a sole carbon source. The strain B1B grown on biphenyl showed a good degradation activity for 4,4'-dichlorobiphenyl (DCBp) while strains P166 and T104 gave about 25% of B1B activity. Induction of degradation by cymene, limonene and terpine was hardly detected by the resting cell assay technique. This appeared to be due to relatively lower induction effect of these terpenoids compared with biphenyl. However, a subsequent GC analysis showed that the congener could be removed up to 30% by the resting cells of T104 grown on the terpenoids. This indicates that terpenoids, widely distributed in nature, could be utilized as both growth and/or inducer substrate for PCB biodegradation.

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Inhibitory Effects of EGCG on the Dopaminergic Neurons

  • Heo, Tag;Jang, Su-Jeong;Kim, Song-Hee;Jeong, Han-Seong;Park, Jong-Seong
    • Biomedical Science Letters
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    • v.15 no.2
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    • pp.127-133
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    • 2009
  • This study was designed to investigate the effects of high concentration of (-)-epigallocatechin-3-gallate(EGCG) on the neuronal activity of rat substantia nigra dopaminergic neurons. Sprague-Dawley rats aged 14 to 16 days were decapitated under ether anesthesia. After treatment with pronase and thermolysin, the dissociated dopaminergic neurons were transferred into a chamber on an inverted microscope. Spontaneous action potentials and potassium currents were recorded by standard patch-clamp techniques under current and voltage-clamp modes respectively. 18 dopaminergic neurons(80%) revealed inhibitory responses to 40 and 100 ${\mu}M$ of EGCG and 4 neurons(20%) did not respond to EGCG. The spike frequency and resting membrane potential of these cells were decreased by EGCG. The amplitude of afterhyperpolarization was increased by EGCG. Whole potassium currents of dopaminergic neurons were increased by EGCG(n=10). These experimental results suggest that high concentration EGCG decreases the neuronal activity of the dopaminergic neurons by altering the resting membrane potential and afterhyperpolarization.

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Effects of Sphingosine-1-phosphate on Vestibular Nuclear Neurons

  • Lee, Jae-Hyuk;Jang, Su-Jeong;Kim, Song-Hee;Jeong, Han-Seong;Park, Jong-Seong
    • Biomedical Science Letters
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    • v.16 no.1
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    • pp.46-52
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    • 2010
  • This study was designed to investigate the effects of sphingosine-1-phosphate on the neuronal activity of rat medial vestibular nuclear neurons. Sprague-Dawley rats aged 14 to 16 days were decapitated under ether anesthesia. After treatment with pronase and thermolysin, the dissociated medial vestibular nuclear neurons were transferred into a chamber on an inverted microscope. Spontaneous action potentials and potassium currents were recorded by standard patch-clamp techniques under current and voltage-clamp modes respectively. 15 medial vestibular nuclear neurons revealed excitatory responses to 1 and $5\;{\mu}M$ of sphingosine-1-phosphate. The spike frequency and resting membrane potential of these cells were increased by sphingosine-1-phosphate. The amplitude of afterhyperpolarization was decreased by sphingosine-1-phosphate. Whole potassium currents of medial vestibular nuclear neurons were decreased by sphingosine-1-phosphate (n=12). Sphingosine-1-phosphate did not affect the charybdotoxin-treated potassium currents. These experimental results suggest that sphingosine-1-phosphate increases the neuronal activity of the medial vestibular nuclear neurons by altering the resting membrane potential and afterhyperpolarization.

Production of Ascorbic acid-2-phosphate from Ascorbic acid by Pseudomonas sp.. (Pseudomonas sp.에 의한 Ascorbic acid로부터 Ascorbic acid-2-phosphate의 생산)

  • 권기성;이상협;방원기
    • Microbiology and Biotechnology Letters
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    • v.28 no.1
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    • pp.33-38
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    • 2000
  • In order to produce ascorbic acid-2-phosphate from ascorbic acid, bacteria capable of transforming ascorbic acid to ascorbic acid-2-phosphate were isolated from soils and the stock cultures in our laboratory. Among them, a newly isolated bacterium LSH-3 having the best ability of producing ascorbic acid-2-phosphate was selected and partially identified as Pseudomonas sp. The optimum conditions for the production of ascorbic acid-2-phosphate from ascorbic acid and using its resting cells as the source os enzyme were investigated. The results were summarized as follows: The optimum cultivation time and the cell weight for the production of ascorbic acid-2-phosphate was 14 hours and 100g/I(wet weight), respectively. And 0.1%(v/v) Trition X-100 was the most effective surfactant. The optimum concentrations of ascorbic acid and pyrophosphate were 400mM and 500mM, respectively, which led to produce 14.54g/I of ascorbic acid-2-phosphate. The most effective buffer was 50mM sodium acetate. The optimum pH and temperature were 4.5 and $40^{\circ}C$, respectively. Under the above conditions, 17.71 g/I of ascorbic acid-2-phosphate was produced from ascorbic acid after 32 hour-incubation, which corresponded to 17.5% of conversion rate based on ascorbic acid.

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Optimization of Ascorbic Acid-2-Phosphate Production from Ascorbic Acid Using Resting Cell of Brevundimonas diminuta

  • Shin, Woo-Jung;Kim, Byung-Yong;Bang, Won-Gi
    • Journal of Microbiology and Biotechnology
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    • v.17 no.5
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    • pp.769-773
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    • 2007
  • With the aim to produce ascorbic acid-2-phosphate(AsA-2-P) from L-ascorbic acid(AsA, Vitamin C), nine bacteria conferring the ability to transform AsA to AsA-2-P were isolated from soil samples alongside known strains from culture collections. Most isolates were classified to the genus Brevundimonas by 16S phylogenetic analysis. Among them, Brevundimonas diminuta KACC 10306 was selected as the experimental strain because of its the highest productivity of AsA-2-P. The optimum set of conditions for the AsA-2-P production from AsA using resting cells as the source of the enzyme was also investigated. The optimum cultivation time was 16 h and the cell concentration was 120g/l(wet weight). The optimum concentrations of AsA and pyrophosphate were 550mM and 450mM, respectively. The most effective buffer was 50mM sodium formate. The optimum pH was 4.5 and temperature was $40^{\circ}C$. Under the above conditions, 27.5g/l of AsA-2-P was produced from AsA after 36 h of incubation, which corresponded to a 19.7% conversion efficiency based on the initial concentration of AsA.

Effects of Ethanol Extract of Rehmanniae Radix(RREE) on Resting and Action Potential of Rabbit Sinoatrial Node and Papillary Muscle (Rehmanniae Radix에 의한 가토 동방결절 및 유두근의 활동전압의 변동)

  • Hwang, Bo-Sang;Koh, Sang-Don;Shin, Hong-Kee;Kim, Kee-Soon
    • The Korean Journal of Physiology
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    • v.19 no.2
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    • pp.127-137
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    • 1985
  • The present study was undertaken in order to investigate effect of ethanol extract of Rehmanniae radix(RREE) on electrophysiology of sinus node and papillary muscle. Rehmanniae radix is a herbal medicine which has been known to have diuretic, antipyretic, hemopoietic and cardiotonic effects. Action potentials were recorded by means of glass capillary microelectrode(technique) in rabbit sinoatrial nodal cells and papillary muscle cells which were superperfused with either tyrode solution or tyrode solutions containing different amount of RREE. The results obtained were as follows ; 1) In both central and peripheral nodal cells maximum diastolic potential (MDP) and amplitude of action potential (APA) were not affected by RREE. 2) Action potential duration as expressed $APD_{60}$(time to 60% repolarization) of central and peripheral pacemaker cells were significantly prolonged following perfusion with tyrode solution containing 0.1% RREE. 3) The rates of spontaneous firing from central pecemaker cell were decreased by RREE at concentration of 0.05% and 0. 1% while spontaneous rhythm of perinodal cell was decreased by 0.1% RREE. 4) The action potential duration of papillary muscle as expressed $APD_{60}$ were prolonged by 0.1% RREE.

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