• The Korean Journal of Pharmacology
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• v.26 no.2
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• pp.127-133
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• 1990

Dibutyryl-cyclic AMP (db-cAMP) and forskolin were used to investigate vasodilating mechanism of cAMP in rabbit aorta. Db-cAMP and forskolin inhibited the development of contractile tension induced by norepinephrine (NE) concentration-dependently. However, high $K{^+}-induced$ contractile tension was inhibited less effectively by db-cAMP and forskolin. Db-cAMP and forskolin inhibited $^{45}Ca^{2+}$ uptake increased by NE. Forskolin seemed to inhibit $^{45}Ca^{2+}$ uptake increased by high $K{^+}$, but this inhibition was not significant statistically. Db-cAMP inhibited $Ca^{2+}-transient$ contraction by NE in $Ca^{2+}-free$ solution. In conclusion, it seems that cAMP blocks $Ca^{2+}$ influx through receptor operated $Ca^{2+}$ channels (ROCs), but that the effect of cAMP on $Ca^{2+}$ influx through voltage gated $Ca^{2+}$ channels (VGCs) is not clear in this experiment. Furthermore, cAMP is likely to inhibit calcium release from the intracellular stores.

• The Korean Journal of Pharmacology
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• v.29 no.2
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• pp.165-174
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• 1993

$Ca^{2+}$ is an important regulator of neurite elongation and growth cone movements but the mechanism(s) mediating these $Ca^{2+}-dependent$ effects is unclear. Since cytoskeletal proteins are rapidly degraded by $Ca^{2+}-dependent $ proteinases (calpains) in vitro and in vivo, we investigated whether $Ca^{2+}-induced$ pruning or regression of neuronal processes is mediated by calpains. Isolated hippocampal pyramidal-like neurons were cultured and the ability of the membrane-permeable calpain inhibitors EST (etyl (+)-(2S,3S)-3-[(S)-methyl-1-(3-methlbutylcarbamoyl)-butylcarbamoyl]2-oxiranecarboxylate) and MDL28170 (carbobenzoxyl-Val-Phe-H) to block the $Ca^{2+}$ ionophore A23187-induced suppression in neurite outgrowth was investigated. Addition of 100 nM A23187 to the culture medium resulted in a retraction of dendrites without altering axonal elongation. The addition of 300 nM A23187 to the culture medium resulted in a signiciant decrease in the rate of axonal elongation as well as a retraction of dendritic processes. Administration of EST $(5\;or\;20{\mu}M)$ to the culture medium completely blocked the pruning effect of 100 nM A23187 on dendrites and of 300 nM A23187 on axons, while EST alone did not significantly affect neurite outgrowth rate. MDL 28170 $(20\;{\mu}M)$ showed the same effect as EST in preventing ionophore-induced pruning of dendrites and axons at 100 nM and 300 nM concentrations, respectively, of A23187. EST $(20\;{\mu}M)$ did not block the A23187-induced rise of $[Ca^{2+}]_{i}$ as measured with fura-2. These results show that $Ca^{2+}-induced$ pruning of neurites in isolated hippocampal pyramidal neurons is mediated by calpains.

• The Korean Journal of Pharmacology
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• v.24 no.1
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• pp.71-81
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• 1988

Glutamate and aspartate may evoke an increase in membrane permeability to monovalent cations and $Ca^{++}$. However, it is uncertain whether $Ca^{++}$ influx is mediated by voltage dependent $Ca^{++}$ channels or by excitatory amino acid activated channels. In addition, the influences of excitatory amino acids on $Ca^{++}$ uptake by neuronal tissues as well as the responses of their actions to extracellular $Mg^{++}$ concentration are different. $K^{+}$ induced $Ca^{++}$ uptake by synaptosomes was dependent on extracellular $Mg^{++}$ up to 5 mM and at concentration of 10 mM, $Ca^{++}$ influx was rather reduced. In $Na^{+}$ rich media, glutamate-and aspartate-induced $Ca^{++}$ uptake was increased by $Mg^{++}$ in a dose independent manner. However, the response for NMDA was inhibited by $Mg^{++}$ at concentrations above 2 mM. $K^+$-and glutamate-induced $Ca^{++}$ influx s were inhibited by 2,4-dinitrophenol, chlorprom-azine and verapamil but not by tetraethylammonium chloride. Tetrodotoxin effectively inhibited the action of glutamate but did not affect that of $K^+$. The response for MNDA was inhibited by 2, 4-dinitrophenol and tetrodotoxin, slightly inhibited by verapamil, and not affected by tetraethylammonium chloride. In $Na^{++}$ rich medium, depolarizing action of glutamate, aspartate and MNDA on synaptosomes was not demonstrated, whereas these agents stimulated $Ca^{++}$ uptake and caused $Ca^{++}$ influx induced depolarization at mitochondria. On the other hand, the activities of synaptosomal ATPases were not affected by excitatory amino acids at 5 mM. The results suggest that glutamate or NMDA induced $Ca^{++}$ influx at synaptosomes exhibits different responses for extracellular $Mg^{++}$ Ex citatory amino acids induced $Ca^{++}$ influx at synaptosomes may be associated with increased permeability of membrane for $Na^{++}$ and $Ca^{++}$ except $K^{++}$ and membrane depolarization due to increased ionic permeability.

• The Korean Journal of Pharmacology
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• v.29 no.2
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• pp.195-202
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• 1993

Oxidative modification of cellular proteins and lipids may play a role in the development of diabetic complications. Diabetic cardiomyopathy has been suggested to be caused by the intracellular $Ca^{2+}$ overload in the myocardium, which is partly due to the defect of calcium transport of the cardiac sarcoplasmic reticulum (SR). In the present study, the possible mechanism of the functional defect of cardiac SR in diabetic rats was studied. Both of the maximal $Ca^{2+}$ uptake and the affinity for $Ca^{2+}$ were decreased in the diabetic rat SR in comparison with the control. To investigate whether the functional defect of the cardiac SR in streptozotocin-induced diabetic rat is associated with the oxidative changes of cardiac SR proteins, the carbonyl group content and glycohemoglobin levels were determined. The increase in carbonyl group content of cardiac SR (2.30 nmols/mg protein, DM; 1.78, control) and in glycohemoglobin level $(13{\sim}17%,\;DM;\;3{\sim}5%,\;control)$ were observed in the diabetics. The extent of increase in calcium transport by phospholamban phosphorylation was greater in the diabetic cardiac SR membranes than that in the control. The phosphorylation levels of phospholamban, as determined by SDS-PAGE and autoradiography with $[{\gamma}^{32}P]ATP$, were increased in diabetic cardiac SR. These results suggest that the impaired cardiac SR function in diabetic rat could be a consequence of the less-phosphorylation of phospholamban in the basal state, which is partly due to the depleted norepinephrine stores in the heart. Furthermore, the oxidative damages in cardiac SR membranes might be one of the additional factors leading to the diabetic cardiomyopathy.

• Journal of Yeungnam Medical Science
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• v.11 no.2
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• pp.363-374
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• 1994

The purpose of this study was to investigate the characteristics or the potassium channels existing in the rat urinary bladders. Smooth muscle strips of rat detrusor urinae were examined by isometric myography. Relaxation responses of detrusor muscle strips to the three potassium channel openers pinacidil, a cyanoguanidine derivative, BRL 38227, a benzopyran derivative and RP 52891, a tertrahydrothiopyran derivative were examined. The potassium channel openers reduced the basal tone, and the rank order of potency was RP 52891>pincidil>BRL 38227. Procaine, an inhibitor of the voltage-sensitive potassium channel tended to increase the basal tone, but it did not affect the relaxant effects of the calcium-activated potassium channel opener did not antagonize the relaxant effects, but it reduced the Emax of RP 52891 and BRL 38227. Glibenclamide, an inhibitor of the ATP-sensitive potassium channel, antagonized the relaxant effects of pinacidil, RP 52891 and BRL 38227 reducing the Emax of RP 52891 and BRl 38227. Galanin which inhibits secretion of insulin through opening the ATP-sensitive potassium channels in pancreatic ${\beta}$-cells rather increased the basal tone of the isolated detrusor strips. These results suggest that the urinary bladder of the rat has mainly the ATP-sensitive, glibenclamide sensitive potassium channel, which is a different type from that in the pancreatic ${\beta}$-islet cells..

• Environmental Analysis Health and Toxicology
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• v.18 no.2
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• pp.89-94
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• 2003

Tributyltin (TBT) used world-wide in antifouling paints for ships is a widespread environmental pollutant and cause reproductive organs atrophy in rodents. At low doses, antiproliferative modes of action have been shown to be involved, whereas at higher doses apoptosis seems to be the mechanism of toxicity in reproductive organs by TBT. In this study, we investigated that the mechanisms underlying DNA fragmentation induced by TBT in the rat leyding cell line, R2C. Effects of TBT on intracellular Ca$\^$2+/ level and reactive oxygen species (ROS) were investigated in R2C cells by fluorescence detector. TBT significantly induced intracellular Ca$\^$2+/ level in a time-dependent manner. The rise in intracellular Ca$\^$2+/ level was followed by a time-dependent generation of reactive oxygen species (ROS) at the cytosol level. Simultaneously, TBT induced the release of cytochrome c from the mitochondrial membrane into the cytosol. Furthermore, ROS production and the release of cytochrome c were reduced by BAPTA, an intracellular Ca$\^$2+/ chelator, indicating the important role of Ca$\^$2+/ in R2C during these early intracellular events. In addition, Z-DEVD FMK, a caspase-3 inhibitor, decreased apoptosis by TBT. Taken together, the present results indicated that the apoptotic pathway by TBT might start with an increase in intracellular Ca$\^$2+/ level, continues with release of ROS and cytochrome c from mitochondria, activation of caspases,and finally results in DNA fragmentation.

• Proceedings of the Korean Society of Applied Pharmacology
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• 1994.04a
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• pp.274-274
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• 1994

본 연구에서는 토끼신장 근위세뇨관 일차배양세포에서 브라디키닌의 생리작용이 phospholipase D (PLD)에 의해 매개되는지를 살펴 보기위해 PLD 효소반응의 특이한 성질인 transphosphatidylation 반응의 생성물인 phosphatidylethanol (PEth) 의 세포내 양을 측정함으로 PLD 효소의 관련성을 규명할 수 있었다. 시간경과에 따른 phosphatidic acid (PA) 및 diacylglycerol (DAG) 의 생성을 살펴본 결과 PA가 DAG보다 먼저 생성되어 최고치 (30초)에 도달하였고 DAG는 1분이후부터 5분까지 서서히 생성되는 양상을 나타내었다. 또한 0.5에서 5％까지의 에탄올 존재하에 PA 및 PE소 생성량을 비교해본 결과 에탄올량이 증가함에 따라 PA는 감소하는 반민 PEth 의 생성은 계속 증가하였다. 한편 브라디키닌 농도 변화 실험에서는 브라디키닌농도가 증가함에 따라 PA 및 PEth 둘다 생성이 증가되었다. 이러한 결과로부터 토끼신장 근위세뇨관 세포막에 존재하는 브라디키닌수용체는 브라디키닌에 의해 activation 시 PLD를 직접적으로 활성화시켜 그들의 작용을 세포내로 전달한다는 사실을 알 수 있었다. 또한 PLD 효소활성의 activator로 수용체효능 제외에 칼슘이온, protein kinase C (PKC) 등이 몇몇 다른 실험에 의해 밝혀져 있고, G protein 역시 PLD 효소 활성을 조절하는 역할이 있음이 알려졌다. calcium ionophore 및 칼슘채널길항제인 verapamil을 이용한 실험에서 우리는 브라디키닌의 PLD 활성화는 칼슘이온에 의존적인 경로 및 비의존적인 경로가 같이 존재함을 알수 있었다. 또한 브라디키닌의 PLD 활성화기전이 PKC 의존적인지를 살펴보기위해 PKC activator(PMA) 및 inhibitor (staurosporine)를 이용한 실험에서 브라디키닌은 신장세포에서 PKC를 통하여 PLD를 활성화시킴으로 신호전달을 하는 것으로 추측되었다. 마지막으로 가수분해안되는 G protein 유도체인 GTPrS 및 G protein 활성물질 NaF, 백일해독소등을 이용한 실험에서 G protein 의 PLD 조절활성을 확인할 수 있었다.

• Journal of Life Science
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• v.13 no.6
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• pp.924-932
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• 2003

Epigallocatechin-3 Gallate (EGCG), a major constituent of green tea, has attracted increasing interest because of its many reported health benefits. Here we demonstrate for the first time that EGCG stimulates phospholipase D (PLD) activity in U87 human astroglioma cells. EGCG-induced PLD activation was abolished by the phospholipase C (PLC) inhibitor and a lipase inactive PLC-\gama1$ mutant, and was dependent on intracellular $Ca^{ 2+}$, and possibly involved $Ca^{ 2+}$ calmodulin-dependent protein kinase II (CaM kinase II). Interestingly, EGCG induced translocation of PLC-\gama1$ from the cytosol to the membrane and PLC-\gama1$interaction with PLD1. Taken together, these results demonstrate for the first time that in human astroglioma cells, EGCG regulates PLD activity via a signaling pathway involving a PLC-\gama1$ (inositol 1,4,5-trisphosphate-$Ca^{ 2+}$)-CaM kinase II-PLD pathway.

• Resources Recycling
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• v.19 no.3
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• pp.62-70
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• 2010

As the demand for paper continues to grow and the importance of recycled paper, white ledgar(WL) and old newspaper pulp(ONP), continuously increase. In addition, usage of recycled paper is essential in terms of forest conservation and environmental protection issues. However, optical and mechanical properties of recycled paper have some drawbacks than regular paper's properties that is indispensable. In order to complement these problems of recycled paper, precipitated calcium carbonate (PCC) was synthesized by the In-situ process with a recycled pulp. Depending on the size of PCC is divided into 2 types, $0.01{\mu}m{\sim}0.09{\mu}m$ colloid type ultra-fine particle and $0.1{\mu}m{\sim}0.9{\mu}m$ cubic type particles. In this study, we analyze how the different shape and size of precipitated calcium carbonate affects in the optical and mechanical properties of the recycled paper used as a filler. Furthermore, we mixed with chemical pulp for overcome reduce of mechanical properties, without using other chemicals, when we use PCC as a filler. The results has the possibility to meet in GR excellent recycling certification mark, standard was proposed.

• Journal of Life Science
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• v.25 no.1
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• pp.1-7
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• 2015

Cyclin D is a member of the cyclin protein family, which plays a critical role as a core member of the mammalian cell cycle machinery. D-type cyclins (D1, D2, and D3) bind to and activate the cyclin-dependent kinases 4 and 6, which can then phosphorylate the retinoblastoma tumor suppressor gene products. This phosphorylation in turn leads to release or derepression of E2F transcription factors that promote progression from the G1 to S phase of the cell cycle. Among the D-type cyclins, cyclin D3 encoded by the CCND3 gene is one of the least well studied. In the present study, we have investigated the biochemistry of the proteolytic mechanism that leads to loss of cyclin D3 protein. Treatment of human prostate carcinoma PC-3-M cells with lovastatin and actinomycin D resulted in a loss of cyclin D3 protein that was completely reversible by the peptide aldehyde calpain inhibitor, LLnL. Additionally, using inhibitors for various proteolytic systems, we show that degradation of cyclin D3 protein involves the $Ca^{2+}$-activated neutral protease calpain. Moreover, the half-life of cyclin D3 protein half-life increased by at least 10-fold in PC-3M cells in response to the calpain inhibitor. We have also demonstrated that the transient expression of the calpain inhibitor calpastatin increased cyclin D3 protein in serum-starved NIH 3T3 cells. These data suggested that the function of cyclin D3 is regulated by $Ca^{2+}$-dependent protease calpain.