• KSBB Journal
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• v.13 no.4
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• pp.399-403
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• 1998

This study was carried out to optimize the concentration of Ca2+ and pH of fusion medium which affected electrofusion frequency of protoplasts isolated from Nicotiana tabacum L. (cv. BY4) mesophyll cells and callus. The protoplasts were electrofused in the fusion media containing two different Ca2+ concentrations and three different pH regions. Fusion frequency was lower in the fusion medium containing only 13% mannitol as osmotic stabilizer. However, higher degree of fusion frequency (47.3%) was observed in the fusion medium containing 50mM CaCl2 at pH 10.5 than any other conditions. Cell viability was decreased by Ca2+ and high pH treatment in the fusion media, while fusion frequency was increased. It is concluded that Ca2+ is involved in electrofusion of protoplasts.

• Archives of Pharmacal Research
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• v.19 no.4
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• pp.280-285
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• 1996

The sustained release dosage form which delivers melatonin (MT) in a circadian fashion over 8 h is of clinical value for those who have disordered circadian rhythms because of its short halflife. The purpose of this study was to evaluate the gelling properties and release characteristics of alginate beads varying multivalent cationic species $(Al^{+++}, \; Ba^{++}, \; Ca^{++}, \; Mg^{++}, \; Fe^{+++}, \; Zn^{++})$. The surface morphologies of Ca- and Ba-alginate beads were also studied using scanning electron microscope (SEM). MT, an indole amide pineal hormone was used as a model drug. The $Ca^{++}, \; Ba^{++}, \; Zn^{++}, \; Al^{++}\; and\; Fe^{+++}\; ions\; except\; Mg^{++}$ induced gelling of sodium alginate. The strength of multivalent cationic alginate beads was as follows: $Al^{+++}\llFe^{+++} the induced hydrogel beads were very fragile and less spherical. Fe-alginate beads were also fragile but stronger compared to Al-alginate beads. Ba-alginate beads had a similar gelling strength but was less spherical when compared to Ca-alginate beads. Zn-alginate beads were weaker than Ca- and Ba-alginate beads. Very crude and rough crystals of Ba- and Ca-alginate beads at higher magnifications were observed. However, the type and shape of rough crystals of Ba- and Ca-alginate beads were quite different. No significant differences in release profiles from MT-loaded multivalent cationic alginate beads were observed in the gastric fluid. Most drugs were continuously released upto 80% for 5 h, mainly governed by the passive diffusion without swelling and disintegrating the alginate beads. In the intestinal fluid, there was a significant difference iq the release profiles of MT-loaded multivalent cationic alginate beads. The release rate of Ca-alginate beads was faster when compared to other multivalent cationic alginate beads and was completed for 3 h. Ba-alginate beads had a very long lag time (7 h) and then rapidly released thereafter. MT was continuously released from Feand Zn-alginate beads with initial burstout release. It is assumed that the different release rofiles of multivalent cationic alginate beads resulted from forces of swelling and disintegration of alginate beads in addition to passive diffusion, depending on types of multivalent ions, gelling strength and drug solubility. It was estimated that 0.2M $CaCl_2$ concentration was optimal in terms of trapping efficiency of MT and gelling strength of Ca-alginate beads. In the gastric fluid, Ca-alginate beads gelled at 0.2 M $CaCl_2$ concentration had higher bead strength, resulting in the most retarded release when compared to other concentrations. In the intestinal fluid, the decreased release of Ca-alginate beads prepared at 0.2 M $CaCl_2$ concentration was also observed. However, release profiles of Ca-alginate beads were quite similar regardless of $CaCl_2$ concentration. Either too low or high $CaCl_2$ concentrations may not be useful for gelling and curing of alginate beads. Optimal $CaCl_2$ concentrations must be decided in terms of trapping efficiency and release and profiles of drug followed by curing time and gelling strength of alginate beads.

• Molecules and Cells
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• v.44 no.2
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• pp.88-100
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• 2021

Anoctamin 6/TMEM16F (ANO6) is a dual-function protein with Ca2+-activated ion channel and Ca2+-activated phospholipid scramblase activities, requiring a high intracellular Ca2+ concentration (e.g., half-maximal effective Ca2+ concentration [EC50] of [Ca2+]i > 10 μM), and strong and sustained depolarization above 0 mV. Structural comparison with Anoctamin 1/TMEM16A (ANO1), a canonical Ca2+-activated chloride channel exhibiting higher Ca2+ sensitivity (EC50 of 1 μM) than ANO6, suggested that a homologous Ca2+-transferring site in the N-terminal domain (Nt) might be responsible for the differential Ca2+ sensitivity and kinetics of activation between ANO6 and ANO1. To elucidate the role of the putative Ca2+-transferring reservoir in the Nt (Nt-CaRes), we constructed an ANO6-1-6 chimera in which Nt-CaRes was replaced with the corresponding domain of ANO1. ANO6-1-6 showed higher sensitivity to Ca2+ than ANO6. However, neither the speed of activation nor the voltage-dependence differed between ANO6 and ANO6-1-6. Molecular dynamics simulation revealed a reduced Ca2+ interaction with Nt-CaRes in ANO6 than ANO6-1-6. Moreover, mutations on potentially Ca2+-interacting acidic amino acids in ANO6 Nt-CaRes resulted in reduced Ca2+ sensitivity, implying direct interactions of Ca2+ with these residues. Based on these results, we cautiously suggest that the net charge of Nt-CaRes is responsible for the difference in Ca2+ sensitivity between ANO1 and ANO6.

• Journal of Embryo Transfer
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• v.16 no.2
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• pp.79-89
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• 2001

Steroid hormones control the expression of many cellular regulators, and a role thor estrogen in mouse oocytes has been well documented. The preovulatory $E_2$increment is generally accepted as the endocrine process regulating induction of in vivo oocyte maturation To address whether the activity of the T-type $Ca^{2+}$ channel is altered by 17 beta-estradiol ( $E_2$), we examined the actions of $E_2$on the calcium channel of mouse oocytes and early embryos. Oocrtes were collected from the oviduct of mice treated with pregnant mare's serum gonadotropin (PMSG) and human choronic gonadotropin (hCG). Whole cell voltage clamp technique and confocal microscopy were used to examine that $E_2$increase intracellular $Ca^{2+}$ concentration ([C $a^{2+}$]$_{i}$ ) via voltage dependent $Ca^{2+}$ channel (VDC) and estrogen receptor (FSR), and $E_2$concentration by the use of radioimmunoassay (RIA) were examined in mouse. The results obtained were as follows: The peak of $Ca^{2+}$ current induced by $E_2$increased 122% to 1.50$\pm$0.03 nA from 1.23$\pm$0.21 nA (n=15) in the presence of 5 mM extracellular $Ca^{2+}$ concentration ([C $a^{2+}$]$_{o}$ ). The increased $Ca^{2+}$ current was temporally associated with $Ca^{2+}$ transients. The intracellular $Ca^{2+}$ level increased 207%~30 s following the addition of 1${\mu}{\textrm}{m}$ $E_2$(relative fluorescence intensity: 836.4$\pm$131.2 for control, n=10, 1736.4$\pm$192.0 in the presence of $E_2$, n=10). $E_2$increased amplitude of $Ca^{2+}$ current and [C $a^{2+}$]$_{i}$ . $E_2$-induced $Ca^{2+}$ current and $E_2$concentration in blood were showed difference on the stage of embryo. These results suggest that $E_2$modulate $Ca^{2+}$ channel to increase $Ca^{2+}$ influx.$Ca^{2+}$ influx.

• 한국생물공학회:학술대회논문집
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• 2000.11a
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• pp.167-170
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• 2000

Proteolytic hydrolysis is one of the main enzymatic reaction of waste activated sludge (WAS) digestion. Pretense excreted from Bacillus stearothermophilus (ATCC 31197) showed optimum temperature of $75^{\circ}C$ for maxium heat stable proteolytic activity against azo casein. The dependency of $Ca^{2+}$, $Zn^{2+}$ on heat stability of proteolytic enzymes were measured with various concentrations. It was shown that $Ca^{2+}$ ion enhanced heat stability of these enzymes. Then thermophilic aerobic digestion (TAD) was performed using B. sterothermophilus with the addition of divalent ions. Performance of TAD process with ATCC 31197 activated by $Ca^{2+}$, $Zn^{2+}$ions in terms of dissolved organic carbon (DOC) concentration, extracellular protein concentration, and scanning electrion microscopy (SEM) analysis. The best result of protein reduction concentration in digestion test was obtained with the addition of 2 mM $Ca^{2+}$ ion.

• Journal of Environmental Science International
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• v.9 no.6
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• pp.505-509
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• 2000

The purpose of this study was to evaluate the effects of $Ca(OH)_2$ and $CO_2$ additions on the corrosion of metal coupons(ductile iron, galvanized steel, copper and stainless steel). Corrosion rate and released metal ion concentration of ductile iron and galvanized steel decreased by adjusting alkalinity, calcium hardness and pH with $Ca(OH)_2$ & $CO_2$ additions on copper and stainless steel were less than those on ductile iron and galvanized steel. When ductile iron coupon was exposed to water treated with Ca(OH)$_2$&$CO_2$, additions, the main components of corrosion product formed on its surface were $CaCO_3$ and $Fe_2 O_3 or Fe_2 O_4$ which often reduce the corrosion rate by prohibiting oxygen transport to the metal surface.

• Korean Journal of Food Science and Technology
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• v.32 no.2
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• pp.402-409
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• 2000

To investigate the textural and physical characteristics of WSF(whole soybean flour) tofu affected by coagulant and its concentration, $CaCl_2$, $CaSO_4$, $GDL(glucono-{\delta}\;lactone)$, $MgCl_2$, and some mixed coagulants were used in this study. Yields of WSF tofu coagulated with $CaSO_4$ and GDL were ranged 4.3-4.5(g/g WSF), but common tofu was 2.2-3.0(g/g soybean). As the water addition ratio increased, L and a value were increased while heating time increased, b value increased. L value of WSF tofu was lower and b value was higher than conventional tofu and 4 commercial tofus. Kinds of coagulant and its concentration significantly affected to textural properties of WSF tofu. As the concentration of coagulant increase, the hardness increased in most all coagulants. WSF tofu coagulated with $CaSO_4$ and GDL recorded low hardness and adhesiveness, and high springiness among the used coagulants at the same concentration. As the hardness increased, the gumminess and chewiness increased in most all coagulants. As coagulation temperature and molding pressure increase, hardness also increased. WSF tofu coagulated with 0.3% $CaSO_4+GDL$ was the most similar in the textural properties with conventional tofu and 4 commercial tofus. WSF tofu coagulated with 0.3% of $CaSO_4+GDL$ at $85^{\circ}C$, 10 times water addition, 5min. heating and $25.0g/cm^2$ molding pressure recorded the highest score in the sensory evaluation.

• The Korean Journal of Physiology
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• v.26 no.2
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• pp.129-136
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• 1992

The effects of $Cd^{2+}$ on spontaneous contraction, and the contractures induced by $0mM\;Na^+,\;60mM\;K^+\;and\;10^{-6}\;M$ acetylcholine, 1mM caffeine were studied in order to elucidate diverse actions of $Cd^{2+}$ on the $Ca^{2+}$ mobilization related with contractility in the antral circular muscle of guinea pig stomach. $Cd^{2+}$ inhibited the spontaneous contraction in a does dependent manner $(10^{-6}\;M\;10^{-4}\;M).\;Cd^{2+}\;(3{\times}10^{-5}M)$ suppressed 60 mM $K^+$ induced contracture composed or a phasic and a tonic response and the increased tonic response by the increased external $Ca^{2+}$ concentration. $Cd^{2+}$ also suppressed acetylcholine induced contracture composed of repetitive phasic and a tonic component and the increased tonic response by the increased external $Ca^{2+}$ concentration. Caffeine in the concentration of 1mM evoked contracture but $Cd^{2+}$ suppressed the contracture. $Cd^{2+}$ suppressed the amplitude of the $Na^+$ tee contracture dose dependently and the amplitude of $Na^+$ free contracture almost decreased to 20% of control amplitude in the concentration of $10^{-4}\;M\;Cd^{2+}$. From the above results, it is suggested that $Cd^{2+}$ may inhibit not only $Ca^{2+}$ influx via voltage sensitive, receptor operated $Ca^{2+}$ channel and Na/ca exchange but also intracellular $Ca^{2+}$ release from the sarcoplasmic reticulum in the antral circular muscle of guinea pig stomach.

• The Korean Journal of Zoology
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• v.15 no.4
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• pp.163-182
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• 1972

The effects of caffeine on the ATPase activity and Ca uptake of the fragmented sarcoplasmic reticulum isolated from rabbit skeletal muscle were studied. The ATPase activity of the heavy fraction (2,000-8,000xG) was stimulated by caffeine while that of other lighter fractions was not. It is suggested that the enhancement of the ATPase by the caffeine treatment. The Ca uptake of the heavy and middle (10,000-20,000xG) fractions was inhibited by caffeine when measured at the medium Ca concentration higher than 200 nmoles/mg protein, while only that of the heavy fraction was inhibited when measured at the Ca concentration below 200 nmoles/mg protein. Experiments with dicumarol suggested that caffeine inhibits the Ca uptake of the mitochondria as well as that of the sarcoplasmic reticulum and that the inhibition of the Ca uptake by caffeine in the low Ca concentration in the heavy fraction is due to the inhibition of the mitochondrial Ca uptake by caffeine. It appeared highly probable that the potentiation of muscle contraction caused by caffeine is solely due to the inhibition of the Ca uptake by and to the release of the accumulated Ca from the sarcoplasmic reticulum.

• Animal Bioscience
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• v.35 no.9
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• pp.1426-1433
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• 2022

Objective: The present research was conducted to evaluate the interactive effect of dietary concentration of calcium (Ca) and 25-hydroxy vitamin D3 (25OHD3) on the performance, blood composition and digestibility of laying hens. Methods: A total of 540 Hy-line brown laying hens aged 61 to 70 weeks were randomly allotted in a 3×3 factorial arrangement, consisting of three levels of 25OHD3 (0, 25, and 50 ㎍/kg) and three levels of Ca (3.5%, 4.0%, and 4.5%). All diets had basal concentration of 3,000 IU/kg of vitamin D3 including the 2,800 kcal/kg of metabolic energy and 16% of crude protein. Results: The results showed that interactive effect (p<0.05) between Ca and 25OHD3 was such that dietary 25OHD3 linearly increased interleukin-6 at all levels of Ca inclusion. Interaction (p<0.05) occurred with the highest parathyroid hormone in laying hens that received dietary concentration of Ca (3.5%) with 25OHD3 (50 ㎍/kg), and Ca (4.0%) with 25OHD3 (50 ㎍/kg). Egg production and egg weight significantly (p<0.05) increased in the 4.5% Ca group compared to the 3.5% to 4.0% Ca groups. Egg shell thickness and tibia bone length also increased (p<0.05) in groups fed a high-Ca diet (4.0% to 4.5%). Phosphorus digestibility significantly (p<0.05) increased along with dietary Ca level. Among the tested 25OHD3 groups, higher (p<0.05) egg production and tibia thickness were present in hens fed 50 ㎍/kg of 25OHD3. Furthermore, Ca digestibility serum Ca and 25OHD3 were significantly increased in group offered 50 ㎍/kg of 25OHD3. Conclusion: The results gathered in this study indicate that dietary concentrations of 4.0% to 4.5% Ca and 50 ㎍/kg 25OHD3 improve the performance of hens from 61 to 70 weeks of age.