• Clinical and Experimental Reproductive Medicine
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• v.19 no.2
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• pp.105-116
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• 1992

The present study was examined to clarify the role of calcium ion as a factor for the maturation of mouse oocytes. Follicles and cumulus-enclosed oocytes were isolated with two sharp needles under a stereomicroscope from female mouse (ICR) ovaries which were treated PMSG 5 IU 45-46 hours previously. Isolated follicles and cumulus-enclosed oocytes were cultured for 14-16 hours in an organ culture system at $37^{\circ}C$, 5% $CO_2$ in air and 100% humudified in incubator. MHBS was the basic medium used from which A23187, verapamil, $NiCl_{2.}$ $6H_2O$ and $LaCl_{3.}$ $7H_2O$ were added depending on the experimental groups. In follicle- or cumulus-enclosed oocytes wre cultured in these differently treated media. Following results were obtained from the present study. 1. The calcium ionophore A23187 directly or indirectly seems to stimulate GVBD of follicle-enclosed mouse oocytes. Increasing concentration of ionophore A23187 1ed to an increase in oocytes degeneration from the cumulus-enclosed mouse oocytes. 2. The organic $Ca^{++}$ channel blocker, verapamil does not induce GVBD of follicle-enclosed mouse oocytes. Specially, higher dose of 1 mM verapamil induced GVBD of follicle-enclosed mouse oocytes. However, cytoplasm of GVBD oocytes in 1 mM verapamil treated groups appeared shrunk. In the cumulus-enclosed oocytes, polar body formation was reduced in verapamil treated groups and degeneration increased. Verapamil inhibit oocyte maturation (polar body formation). 3. The $Ca^{++}$ inhibitor, Nickel ($NiCl_{2.}$ $6H_2O$) inhibits maturation of the follicle-enclosed oocytes. In the cumulus-enclosed oocytes the progression to MII (PB formation) was reduced and degeneration of mouse oocytes increased as the concentration of $Ni^{++}$ increase. The results indicates that nickel act as an inhibitor of calcium. 4. The $Ca^{++}$ inhibitors, Lanthanum ($LaCl_{3.}$ $7H_2O$) has shown different effect from that of nickel. In follicle-enclosed oocytes, 0.01mM lanthanum induced maturation of mouse oocytes. Polar body formation was reduced in the cumulus-enclosed oocytes all lanthanum treated group.

• The Korean Journal of Pharmacology
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• v.32 no.3
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• pp.347-355
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• 1996

We examined the effects of EGTA and verapamil on phorbol ester-and forskolin-stimulated LH releases and $LH{\beta}$ subunit mRNA levels in order to verify the role of extracellular $Ca^{++}$ on PKC- or cAMP-induced increases in LH release and $LH{\beta}$ subunit mRNA levels in cultured anterior pituitary cells of rat. Forskolin-stimulated $LH{\beta}$ subunit mRNA levels as well as LH release were all suppressed by prevention of $Ca^{++}$ mobilization from extracellular environment, after the treatment of EGTA as a $Ca^{++}$ chelator or verapamil as a $Ca^{++}$ channel blocker. PMA-stimulated $LH{\beta}$ subunit mRNA levels were also suppressed by the treatment of EGTA and verapamil, while PMA-induced LH release was not affected. From the present study, it is, therefore, suggested that PKC activation and cAMP elevation all stimulate $LH{\beta}$ subunit mRNA levels and these are extracellular $Ca^{++}$-dependent. However, LH releases by PKC activation and cAMP increase seem to be different each other. LH release by PKC activation is thought to be independent of extracellular $Ca^{++}$. On the other hand, cAMP stimulated-LH release is thought to be dependent on the entry of extracellular $Ca^{++}$.

• The Korean Journal of Pharmacology
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• v.28 no.1
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• pp.91-102
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• 1992

In $Ca^{++}$ containing media, arachidonic acid markedly stimulated superoxide and $H_2O_2$ generation and activated NADPH oxidase. In $Ca^{++}$ free media, stimulatory action of arachidonic acid on NADPH oxidase was not detected. Arachidonic acid-stimulated respiratory burst was inhibited by EGTA, TMB-8, verapamil, diltiazem, nifedipine, dibucaine, lidocaine, CCCP, 2,4-dinitrophenol, sodium arsenate, chlorpromazine, theophylline, $HgCl_2$, PCMB and PCMBSA but not affected by tetrodotoxin, tetraethylammonium chloride and procaine. EGTA almost completely inhibited release of ${\beta}-glucuronidase$ by arachidonic acid and verapamil, CCCP and theophylline slightly inhibited it, whereas dibucaine did not show any significant effect. Arachidonic acid induced $Ca^{++}$ release from intact neutrophils and it was decreased by TMB-8. Arachidonic acid-induced elevation of intracellular free $Ca^{++}$ level was inhibited by EGTA and CCCP and slightly inhibited by TMB-8. Amount of intracellular free $Ca^{++}$ increased by either arachidonic acid plus verapamil or arachidonic acid plus dibucaine was greater than that by arachidonic acid alone. These results suggest that various changes of biochemical events may be implicated in the functional expression in neutrophils activated by arachidonic acid. Arachidonic acid appears to elevate cytosolic free $Ca^{++}$ level by stimulating $Ca^{++}$ release from intracellular $Ca^{++}$ storage sites. During activation of neutrophils, $Ca^{++}$ influx and efflux may be accomplished, simultaneously.

• Journal of Yeungnam Medical Science
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• v.9 no.1
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• pp.75-89
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• 1992

The development of multi drug-resistant tumor cell population is a major problem in the chemotherapy of human cancer. These cells are often cross resistant to unrelated drugs and the precise mechanisms of multidrug resistant phenotype of tumor cells has not been fully elucidated. Cisplatin resistant tumor cell(SNU-1/$Cis_5$) was induced from human stomach cancer cell line(SNU-1) in vitro. Growth profiles of survival cells were observed during 5 days by thiazolyl blue (MTT) assay. To investigate the cross resistance of various anticancer drugs in SNU-1 and SNU-1/$Cis_5$, We compared the value of $IC_{50}$ - drug concentration at 50% survival of control and gained relative resistances (RR). The RR for SNU-1/$Cis_5$ were as follows; vinblastine, > 43.0 ; epirubicin, 22.9 ; dactinomycin, 16.0 ; etoposide, 15.0 ; vincristine, 9.2 ; adriamycin, 5.7 ; aclarubicin, 5.3. But 5-fluorouracil, methotrexate, daunorubicin have not cross resistance with cisplatin. Resistant inhibition values of $10{\mu}M$ verapamil for SNU-1/$Cis_5$ were as follows; vincristine, 13.1 ; epirubicin, 10.0 ; etoposide, 6.3 ; vinblastine, 4.4 ; dactinomycin, 3.6 ; daunorubicin, 2.4. Membrane proteins of 51,400 and 81,300 daltons were identified by radioiodination with SDS-PAGE, which might represented the drug resistance.

• Journal of Yeungnam Medical Science
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• v.5 no.2
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• pp.59-69
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• 1988

This study was designed to investigate the effect of substitution of strontium for calcium on mechanical activity in isolated perfused spontaneously beating rat hearts. The mechanical activity of the hearts of Langendorff's preparation in conditions of low calcium and strontium-substitution for calcium was compared. The effect of norepinephrine and verapamil were also observed in those conditions. The results were as follows : 1. In low calcium, the mechanical activity of the heart preparation was significantly reduced, but when the equimolar strontium was substituted for the reduced calcium, the activity was kept at similar level to the normal condition. 2. When equimolar strontium was substituted for the total calcium in perfusate, the heart preparation stopped its beating, and it was not restored in spite of reperfusion with normal calcium perfusate. 3. Norepinephrine-induced positive inotropic effect was inhibited in low-calcium condition especially with low concentration of norepinephrine, but not in strontium-substitution for calcium. 4. Verapamil reduced the activity of the heart both in low-calcium and strontium-substitution as well as in normal calcium conditions. From above results, it was concluded that strontium served as a substitute of calcium in maintaining mechanical activity and in responsiveness to norepinephrine, and the influx of strontium through cell membrane is inhibited by verapamil as the influx of calcium.

• Journal of Life Science
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• v.17 no.2 s.82
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• pp.174-178
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• 2007

Treatment of malformin A1 is known to increase ethylene production 130% at 4 hr and 56% at 8 hr after treatment in maize root compared to untreated plants. The ethylene production by malformin A1 was maximum level at 4 hr and slowly decreased up to 8 hr. Calcium ion regulators such as A23187 (calcium ionophore) and verapamil (calcium channel blocker) stimulated ethylene production. Treatment of both calcium ion regulators increased about 30% of ethylene production at 4 hr, and 20% at 8 hr. Both calcium ion regulators did not stimulate malformin A1-induced ethylene production at 4 hr as malformin A1 itself did. However, the treatment of calcium ion regulators with malformin A1 maintains the ethylene production for 8 hr. These results suggested that the proper concentration of calcium might need to confer the effect of malformin A1 on the ethylene production. Malformin A1 suppressed the gravitropic curvature of maize root about 58% at 4 hr and 42% at 8 hr compared to control plant. Verapamil inhibited the gravitropic curvature about 54% at 4 hr and 23% at 8 hr compared to control, respectively. But A23187 could not. In addition, verapamil showed more inhibition in malformin A1-induced gravitropic curvature than A23187 in malformin A1 induced. These data suggested that calcium ion regulators affect the malformin A1-induced ethylene production and gravitropic curvature, and give the evidence that calcium ion play an important role in gravitropic curvature in maize root.

• The Korean Journal of Pharmacology
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• v.29 no.1
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• pp.9-22
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• 1993

To evaluate the protective effects of calcium antagonists, oxygen radical scavengers and excitatory amino acid (EAA) antagonist on the ischemic brain damage, we induced in vitro ischemic condition (namely, lack of oxygen and glucose) to rat hippocampal slices. And the degree of ischemic damage was determined by assaying changes in biochemical parameters such as ATP content and lactate ralease, MDA production in the presence or absence of the various drugs. During experimental ischemia for up to 60 min, ATP content was decreased and the amount of lactate release was markedly increased time-dependently. By changing the reaction medium which contained oxygen and glucose those biochemical parameters were recovered. But the recovery was not complete in this experimental condition. In the same ischemic conditions verapamil and vitamine E prevented the decrease of ATP content and the increase of lactate release from the slices. And verapamil and diltiazem decreased MDA release to the reaction medium. Superoxide dismutase (SOD) and MK-801 (as EAA receptor antagonist) protected the decrease of ATP content and reduced MDA release in 20 min ischemic condition, but glutathione affected ATP content and lactate release at the same condition. When oxygen and glucose were resupplied for 20 min after ischemic condition, verapamil showed the protective effect on the changes of ATP content and lactate release, and vitamine E decreased lactate release (at 20 min ischemia) and MDA release (at 60 min ischemia). These results showed that calcium antagonist and vitamine E protect the ischemic biochemical changes from rat hippocampal slices and calcium antagonist is more potent than vitamine E to protect the ischemical brain damege.

• Journal of Pharmaceutical Investigation
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• v.21 no.3
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• pp.179-188
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• 1991

An HPLC assay method of a drug to be applied to the pharmacokinetic studies of the drug should be completely validated. The validation process for an HPLC assay method in a biological sample was discussed using the data obtained from the development of HPLC method for the simultaneous quantitation of verapamil and norverapamil in human serum. The validation criteria included were specificity, linearity, accuracy, precision, sensitivity, recovery, drug stability, and ruggedness of an assay method.

• The Korean Journal of Physiology
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• v.19 no.1
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• pp.1-13
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• 1985

The influences of $Ca^{2+}-antagonists$, verapamil and $Mn^{2+}$, upon the spontaneous electrical activity and contractions were studied in guinea-pig taenia coli. Spontaneous contractions were recorded with force transducer, and spike action potentials were measured extracellularly by use of suction electrode. All experiments were performed in tris-buffered Tyrode solution Which was aerated With 100% $O_2$ and kept at $35^{\circ}C$. The results obtained were as follows : 1) Verapamil suppressed the frequency and amplitude of spontaneous contractions dose dependently, and blocked completely mechanical responses at the concentration of 1 mg/1. 2) The frequency of bursts of spike discharge(bursts frequency) and the number of spikes in a burst(spikes frequency) were reduced in a dose·dependent manner within the concentration range of $10^{-5}$ to $10^{-3}g/l$, and bursts frequency was affected more readily at a low concentration of $10^{-5}g/l$ verapamil. 3) The verapamil_induced suppression of spontaneous contractions in the Tyrode solution containing 1 mM $Ca^{2+}$ was completely antagonized by the addition of extra $Ca^{2+}$ to the Tyrode solution $(2\;m\;MCa^{2+})$. 4) $Mn^{2+}$ suppressed the amplitude of spontaneous contractions, whereas $Mn^{2+}$ accelerated the frequency dose-dependently within the range of low concentrations$(10^{-7}\;to\;10^{-4}\;M\;Mn^{2+})$. 5) The bursts frequency determining frequency of spontaneous contractions was increased in a dose-dependent manner, whereas the spikes frequency known to determine the contractions amplitude was reduced within the range of low concentrations. At a high concentration of 1 mM $Mn^{2+}$, however, all spontaneous contractile responses were blocked simultaneously with the disapperance of electrical activity. 6) The frequency and amplitude of spontaneous contractions altered by $Mn^{2+}$ in 1mM $Ca^{2+}$ Tyrode were increased by extra $Ca^{2+}(2mM)$.

• The Korean Journal of Pharmacology
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• v.26 no.2
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• pp.113-120
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• 1990

Influences of trigger calcium on myocardial contraction from several sources were investigated on the frequency reduction-induced changes of contraction in rat left atria driven by electrical field stimulation. Rat atria elicited characteristic three phase-changes according to frequency reduction: the first rapid rise in twitch tension, the second transient fast decrease in tension and the third maintenance of twitch tension at about 200% of resting tension during high frequency. Caffeine treatment enormously suppressed the frequency reduction-induced twitch tension increase. The atrial contraction during high frequency vanished after verapamil treatment. But, during low frequency, atrial contraction revived in the presence of verapamil. Ouabain treatment and sodium depletion in superfusing solution abolished the characteristic second phase with slow frequency. These results suggest that slow calcium channel is an indispensable calcium entry route and calcium release from sarcoplasmic reticulum is an major source for trigger calcium in cardiac contraction. And sodium-calcium exchange has a modulatory roles in the regualtion of trigger calcium according to the changes of intracellular sodium concentration.