• The Korean Journal of Pharmacology
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• v.31 no.3
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• pp.355-363
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• 1995

It has been well known that the intracellular calcium concentration $([Ca^{2+}]_i)$ in living cell is very sensitive to live or to survive, but the transmembrane system of calcium ion, especially mechanism of calcium ion movement in unexcitable state has been little elucidated. Though many proposed theories for calcium ion transport have been reported, it is still unclear that how could the sustained maintenance in cytosolic calcium level be done in cell. Since one of possible mechanisms of calcium transport may be related to the acetylcholine receptor-linked calcium channel, author performed experiment to elucidate this mechanism of calcium influx related to cholinergic receptor in ml muscarinic receptor-transfected RBL-2H3 cell-line. 1) The effects of carbachol both on calcium ion influx and on the secretion of hexosaminidase were respectively observed in the manner of time-related or concentration-dependent pattern in this model. 2) The effects of several metal cations on calcium transport were shown in carbachol-induced cell-line. 3) Atropine was administered to examine the relationship between cholinergic receptor and calcium ion influx in this model. 4) PMA (Phorbol 12-myristate 13-acetate) or PTx (Pertussis toxin) was respectively administered to examine the secondary mediator which involved pathway of calcium ion movement in carbachol-induced cell-line. The results of this experiments were as follows; 1) Carbachol significantly stimulated both the calcium influx and the secretion of hexosaminidase in the manner of the concentration-dependent pattern. 2) Atropine potently blocked the effects of carbachol in concentration-response manner. 3) Administered metal cations inhibited the calcium influx in carbachol-stimulated this model to the concentration-related pattern. 4) PMA did not inhibit carbachol-induced secretion of hexosaminidase, but blocked the calcium influx in this cell-line. 5) The suppression of carbachol-induced hexosaminidase secretion was shown in PTx-treated cell -line.

• The Korean Journal of Physiology and Pharmacology
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• v.4 no.2
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• pp.113-120
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• 2000

To investigate the mechanism of smooth muscle contraction induced by emptying of intracellular $Ca^{2+}$ stores, we measured isometric contraction and $^{45}Ca^{2+}$ influx. $CaCl_2$ increased $Ca^{2+}$ store emptying- induced contraction in dose-dependent manner, but phospholipase C activity was not affected by the $Ca^{2+}$ store emptying-induced contraction. The contraction was inhibited by voltage-dependent $Ca^{2+}$ channel antagonists dose dependently, but not by TMB-8 (intracellular $Ca^{2+}$ release blocker). Both PKC inhibitors (H-7 and staurosporine) and tyrosine kinase inhibitors (genistein and methyl 2,5-dihydroxycinnamic acid) significantly inhibited the contraction, but calmodulin antagonists (W-7 and trifluoperazine) had no inhibitory effect on the contraction. The combined inhibitory effects of protein kinase inhibitors, H-7 and genistein, together with verapamil were greater than that of each one alone. In $Ca^{2+}$ store-emptied condition, $^{45}Ca^{2+}$ influx was significantly inhibited by verapamil, H-7 or genistein but not by trifluoperazine. However combined inhibitory effects of protein kinase inhibitors, H-7 and genistein, together with verapamil were not observed. Therefore, this kinase pathway may modulate the sensitivity of contractile protein. These results suggest that contraction induced by emptying of intracellular $Ca^{2+}$ stores was mediated by influx of extracellular $Ca^{2+}$ through voltage-dependent $Ca^{2+}$ channel, also protein kinase C and/or tyrosine kinase pathway modulates the $Ca^{2+}$ sensitivity of contractile protein.

• Journal of Microbiology and Biotechnology
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• v.16 no.11
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• pp.1747-1752
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• 2006

Allergic inflammation results from stimulation of ${\beta}$-hexosaminidase secretion, increased calcium influx, and activation of MAPK pathways. Some fractions of Chamaecyparis obtusa decreased secretion of ${\beta}$-hexosaminidase, calcium influx, ROS, and phosphorylation of ERK. These results suggest that Chamaecyparis obtusa would be valuable for development of allergy therapeutics.

• Journal of Microbiology and Biotechnology
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• v.14 no.3
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• pp.520-524
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• 2004

A protein that exhibited a high similarity to a major serine transporter of Escherichia coli, SdaC, was found in Haemophilus injluenzae Rd. Also, $Na^+$-stimulated serine transport activity was detected in the cells. The sdaC of H. injluenzae was cloned and the properties of the transporter were investigated. The activity of serine transport was stimulated by $Na^+$. Uptake of $Na^+$ elicited by L-serine influx into cells was also observed, which supports the idea that L-serine is transported by a mechanism of $Na^+$serine symport. No uptake of $H^+$ elicited by L-serine influx was detected. This result was not consistent with that obtained with the homologous protein, SdaC of E. coli, which uses $H^+$as a coupling cation. The serine transport via the SdaC of H. influenzae was not inhibited by other amino acids such as threonine or D-serine like the SdaC of E. coli. Thus, the SdaC of H. influenzae is a $Na^+$-dependent L-serine specific symporter and an unusual natural mutant. The $K_m$ and the $V_{max}$, value for the serine transport in the SdaC of H. influenzae were $7.6\mu$M and 22.9 nmol/min/mg protein, respectively.

• The Korean Journal of Pharmacology
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• v.32 no.3
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• pp.417-424
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• 1996

The present study was done to examine the involvement of protein kinase C and protein tyrosine kinase in intracellular $Ca^{2+}$ mobilization in C5a-stimulated neutrophils. Although protein kinase C inhibitors, staurosporine and H-7 inhibited intracellular $Ca^{2+}$ release in C5a-stimulated neutrophils, they did not affect $Ca^{2+}$ influx across the plasma membrane and elevation of $[Ca^{2+}]_i$ C5a-induced intracellular $Ca^{2+}$ release and $Ca^{2+}$ influx were inhibited by protein tyrosine kinase inhibitors, genistein and methyl-2,5-dihydroxycinnamate. ADP-evoked elevation of $[Ca^{2+}]_i$ was inhibited by genistein and methyl-2,5-dihydroxycinnamate but was not affectd by staurosporine and H-7. Genistein and methyl-2,5-dihydroxycinnamate reduced the store-regulated $Ca^{2+}$ influx in thapsigargin-treated neutrophils, while the effect of staurosporine and H-7 was not detected. When neutrophils were preincubated wih phorbol 12-myristate 13-acetate, the stimulatory effect of C5a on the elevation of $[Ca^{2+}]_i$ was reduced. These results suggest that protein tyrosine kinase may be involved in control of intracellular $Ca^{2+}$ release and $Ca^{2+}$ influx across the plasma membrane in C5a-activated neutrophils.

• Journal of Microbiology
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• v.41 no.2
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• pp.78-82
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• 2003

We identified two proteins in Haemophilus influenzae Rd that exhibited high similarity to two major serine transporters of Escherichia coli (SstT and SdaC). Then, we investigated serine transport in H. influenzae Rd and detected $Na^{+}$-stimulated L-serine transport activity. The optimum NaCl concentration for this stimulation was about 20 mM. The uptake of $Na^{+}$ by H. influenzae Rd was found to be elicited by L-serine influx, which supports the idea that L-serine is transported by a mechanism of $Na^{+}$/serine symport. No uptake of $H^{+}$ elicited by L-serine influx was detected. $Na^{+}$/serine symport activity was not inhibited by other amino acids such as L-threonine or D-serine. Two distinct Km values were obtained from the kinetic analysis of serine transport. Thus, two serine transport pathways may exist in H. influenzae Rd, and it appears that both systems are stimulated by $Na^{+}$.

• Biomolecules & Therapeutics
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• v.20 no.6
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• pp.550-555
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• 2012

TRPA1 and TRPV1 are members of the TRP superfamily of structurally related, nonselective cation channels. TRPA1 and TRPV1 are often co-expressed in sensory neurons and play an important role in somatosense such as cold, pain, and irritants. The first leaves of Kalopanax pictus Nakai (Araliaceae) have long been used as a culinary ingredient in Korea because of their unique chemesthetic flavor. In this study, we observed the intracellular $Ca^{2+}$ response to cultured cells expressing human TRPA1 (hTRPA1) and human TRPV1 (hTRPV1) by $Ca^{2+}$ imaging analysis to investigate the ability of the first leaves of K. pictus to activate the hTRPA1 and hTRPV1. An 80% ethanol extract of K. pictus (KPEx) increased intracellular $Ca^{2+}$ influx in a response time- and concentration-dependent manner via either hTRPA1 or hTRPV1. KPEx-induced response to hTRPA1 was markedly attenuated by ruthenium red, a general blocker of TRP channels, and HC-030031, a specific antagonist of TRPA1. In addition, the intracellular $Ca^{2+}$ influx attained with KPEx to hTRPV1 was mostly blocked by ruthenium red, and capsazepine, a specific antagonist of TRPV1. These results indicate that KPEx selectively activates both hTRPA1 and hTRPV1, which may provide evidence that the first leaves of K. pictus primarily activate TRPA1 and TRPV1 to induce their unique chemesthetic sense.

• Clinical and Experimental Reproductive Medicine
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• v.18 no.2
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• pp.153-162
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• 1991

The present study was undertaken to clarify the role of calcium ion as a factor for the maturation of follicle-enclosed mouse oocytes. Follicles were isolated with two sharp needles under a stereomicroscope from mouse(ICR) ovaries which were treated PMSG 5 IU 45 hours previously. Isolated follicles were cultured for 14-16 hours in an organ culture system at $37^{\circ}C$, 5% $CO_2$ in air and in a 100% humidified incubator by treatment of hCG, EDTA and $^{45}Ca^{++}$. Culture medium was Modified Hank's Balanced Salt Sol. (MHBS) and addition of hCG (human chorionic gonadotropin) was made into two doses level 0.4 IU and 0.8IU from the stock sol. and also $^{45}Ca^{++}$ was treated in the culture medium. To explain the role of calcium, calcium chelating agent EDTA was treated to the culture of the mouse follicle-enclosed oocytes. Two observations were made in the present study; nucleus phase and $^{45}Ca^{++}$ uptake into the oocyte. HCG induced oocyte maturation in the follicle about two folds as much as the control group, whereas there is no difference in oocyte maturation between 0.4 IU and 0.8 IU of hCG. Optimum level of hCG seems to be 0.4 IU/ml in the mouse follicle culture. HCG stimulated $^{45}Ca^{++}$ uptake into the oocyte of the follicles by two folds. $^{45}Ca^{++}$ uptake in the control group is about 2.5 folds in comparison of the EDTA(1.71mM) treated group. However, calcium uptake in the EDTA treated groups tends to increase depending on the decrease of EDTA concentration. These observations suggest that firstly, hCG stimulates maturation of the oocyte of the follicle, secondly, $Ca^{++}$ influx is induced by hCG and thirdly, $Ca^{++}$ influx by the treatment of EDTA decreases as a dosage-dependent process. This $Ca^{++}$ uptake may take place by the changes of permeability which was induced by hCG treatment. That is, $Ca^{++}$ influx may trigger the resumption of oocyte maturation. It is further necessary in the future study how this $Ca^{++}$ uptake is induced by hCG and increases permeability of the follicle and oocyte.

• The Korean Journal of Mycology
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• v.15 no.4
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• pp.224-230
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• 1987

Effects Of organic compound, photosensitizer and $K^+$ ion influx. On the light-induced ATPase of mitochondria in L. edodes purified by linear sucrose density gradient centrifugation were studied. The mitochondrial ATPase activity was investigated by various wavelength illumination at dark state. The mitochondrial ATPase was activated 139% and 128% by 10m mol dithiothreitol and 0.1m mol quinacrine, respectively. This enzyme also was activated 36% by 0.1m mol phenazine methosulfate as photosensitizer. But, 100 mg oligomycin and 1m mol phlorizin inhibited activity of enzyme to 48% and 45%, respectively. Its optimum wavelength was 690 nm on the effect of $K^+$ ion influx, its optimum pH and temperature were found to be 7.2 and $55^{\circ}C$.

• 대한약리학회:학술대회논문집
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• 2006.10a
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• pp.92.2-92.2
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• 2006