• Journal of Plant Biology
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• v.39 no.3
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• pp.173-177
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• 1996

A novel calcium binding protein (CaBP) was purified to electrophoretic homogeneity from Dunaliella salina. In the course of purification experiment, this CaBP was identified as a monomer and its molecular weight was about 21 kDand isoelectric point (pI) value was about 4.1 using isoelectrofocusing. This CaBP was able to bind Ca2+ even in the pressence of an excess MgCl2 and KCI both in solution. In the SDS-PAGE, the Ca2+-bound form was slower than the Ca2+-free form in the nondenaturing PAGE. This means that the CaBP undergoes conformational change in the Ca2+-bound condition. Furthermore, UV absorption spectrum and fluorescence intensity of this CaBP was investigated. UV absorption peak was appeared at about 258 nm and decreased somewhat in Ca2+-bound condition. In the measurement of fluorescence, maximum intensity was appeared at 303 nm and decreased in Ca2+-bound state, similarly as UV absorption spectrum. These show distinct changes upon Ca2+-binding, which indicate of structural and/or dynamic changes largely reminiscent of other members of the EF-hand Ca2+-binding protein family.

• Journal of Veterinary Clinics
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• v.9 no.2
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• pp.373-390
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• 1992

To assay the fertilizing capacity of domestic animal spermatozoa by hamster test, semen were collected from 13 boars(Duroc. Landrace and Yorkshire) which had been proved to be fertile in the past. then, were preserved in BWW medium or in raw state at 18$^{\circ}C$ or at room temperature. The preserved semen were given each different treatment according to the experimental design and coincubated with zona-free hamster ova for 5 hours. The ova were stained by lacmoid and examined under phase contrast microscope to investigate the rates of ova bound with sperm(sperm binding). ova penetrated by sperm(penetration) and formation of a male pronucleus(pronucleus formation) and also numbers of both bound and penetrated sperm per ovum. Between BWW and TBM medium for boar sperm. no difference in the results of hamster test was obtained. The boar spermatozoa in BWW medium, BWW with caffeine, BWW with heparin, and BWW with both caffeine and heparin showed no difference in the results of hamster test. The boar spermatozoa in BWW medium containing both calcium and RSA showed considerably higher rates of sperm binding, penetration and pronucleus formation as well as higher numbers of both bound and penetrated sperm than those not containing calcium with or without BSA( p<0.01) and also the same results higher than that containing calcium without BSA( p< 0.05). The boar spermatozoa irradiated by X-ray(70 KVP, 20mA) for 3 seconds. then, maintained at 18$^{\circ}C$ for 18 hours showed considerably lower rate of sperm binding than all the other groups including the control and X-ray groups irradiated by smaller dose or maintained for shorter period(p<0.01), and also showed lower number of bound sperm than the other groups(p<0.01, p<0.05). All the control groups of both raw and diluted sperm in BWM medium showed higher rates of sperm binding, penetration and pronucleus formation as well as higher number of penetrated sperm than all the X-ray groups irradiated for 3 seconds(70KVP, 20mA) and maintained for either 3 or 18 hours (p<0.01, p<0.05). At the same time the control groups of diluted sperm showed considerably higher rates of sperm penetration and pronucleus formation than the control group of raw sperm( p<0.01). These results indicates that fertile boar sperm showed considerably lower rates In the results of hamster test, when incubated in the medium without calcium and irradiated by X-ray than when incubated in the medium with calcium and not irradiated by X-ray, respectively, to prove consequently that hamster test would be of great value in assaying the fertilizing capacity of boar spermatozoa.

• Journal of radiological science and technology
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• v.32 no.1
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• pp.69-78
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• 2009

The cardiovascular disease has been known as a common cause of death for a long time in the west. The eating habits of Asia, including Korea, have changed recently, so that this disease is also a problem in Asia now. Annual Report on the Cause of Death Statistics from 1996 to 2006 reported that the cardiovascular disease would become the number one cause of death in the next $5{\sim}10$ years. Therefore we realize that more accurate examination is required. The aim of this study was to investigate the accuracy of Calcium-scoring CT and the relationship between risk factor and quantitative scores of Calcium-scoring CT. Through this study we expect that the national public health will be improved. Seventy patients with chest pain were chosen at random. The patients were undergone both coronary CT antigraphy and Calcium - scoring CT at G hospital in Incheon from February 1 to June 30, 2008. The result of the Calcium-scoring CT showed its usefulness for Ischemic cardiovascular disease, with an accuracy similar to that of exercise/pharmacologic stress or ECG when it is difficult for a patient to exercise due to joint problems, aging or for other reasons.

• Journal of Microbiology and Biotechnology
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• v.14 no.3
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• pp.607-611
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• 2004

Glycinin, one of the predominant storage proteins in soybeans, was examined as to whether it could be used as a calcium-binding mediator after chemical phosphorylation and enzymatic hydrolysis. Glycinin is composed of six subunits. One of the proglycinin subunits $(A_{la}B_{lb})$ was overexpressed in E. coli to obtain nonphosphorylated proteins with homogeneity. To investigate the enhanced calcium-binding properties of the phosphopeptides, the proglycinin was purified, phosphorylated, and hydrolyzed with trypsin. The proglycinin expressed in E. coli was purified by ammonium sulfate precipitation, ion-exchange chromatography, and cryoprecipitation. Chemical phosphorylation by sodium trimetaphosphate was performed to obtain phosphorylated proglycinin. After the phosphorylation, one-dimensional isoelectric focusing gel electroanalysis confirmed the phosphorylation of the proglycinin. The phosphorylated peptides were then hydrolyzed with trypsin, followed by a binding reaction with calcium chloride. The calcium-bound phosphopeptides were finally separated using immobilized metal $(Ca^{2+})$ chromatography. Consequently, a limited tryptic hydrolysate of the isolated phosphopeptides exhibited an enhanced calcium-binding ability, suggesting the potential of glycinin phosphopeptides as a calcium-binding mediator with greater availability.

• Korean Journal of Environmental Agriculture
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• v.16 no.3
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• pp.239-244
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• 1997

To develope more effective extraction methods for paraquat in soil, some modification methods were accomplished in two different types of soil. For extraction of tightly bound-paraquat, conc. HCl 70ml were added with different shaking times, and then $H_2SO_4$ reflux were performed for an hour. In this case, 60 minutes shaking were optimum and recovery were increased more $1.09{\sim}1.50$ folds(84.0% in high clay contents soil, but 96.7% in low clay contents soil) and the long-time consuming step, filtration were easily done, with decreasing filtration time were shorter 4.6 folds(ca. $11{\sim}14min.$). than general paraquat analytical method(ca. $55{\sim}65min.$). And only $H_2O_2$ digestion with different volume and refluxing time resulted in recovery increasing. Nevertheless, considering analyst's safety, 30ml of $H_2O_2$ addition and 30 minutes reflux were regarded as optimum condition. Although, Kjeldahl digestion with $H_2O_2$ showed relatively high recovery, it is not significant statistically. For extraction of loosely bound-paraquat, 0.01, 0.1, 1.0, 10.0M of $NH_4Cl$ and of $CaCl_2$ compared with $1.5{\sim}24hr$ of different shaking time. There were no loosely bound residues of paraquat.

• Asian-Australasian Journal of Animal Sciences
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• v.24 no.10
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• pp.1372-1376
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• 2011

The content of Ca in milk exceeds the typical saturation level of Ca salts, which is necessary for neonate growth. This calcium is distributed between the casein micelles in the colloidal and aqueous phases. Information on the properties of calcium activity in the aqueous phase is limited compared with that on the properties of bound or sequestrated calcium. The objectives of this study were to evaluate the changes in calcium activity in fresh milk using an ion-selective electrode and to assess the relationship between calcium activity and milk production in hot season. Milk samples collected from 10 cows at the National Agricultural Research Center for Kyushu Okinawa Region in June to October (Min-Max: 7.2-$35.2^{\circ}C$, 24.3-100% RH) were analyzed on total calcium concentrations and calcium activity. We observed that the rectal temperature of the cows increased according to elevation of ambient temperature but that the pH of the collected milk ($6.61{\pm}0.01$ (Mean${\pm}$SEM)) was not significantly influenced by rectal and ambient temperature. Total calcium concentrations and calcium activity in fresh milk decreased in July (Min-Max: 21.1-$33.5^{\circ}C$, 48.9-100.0% RH) compared with the values after August (Min-Max: 18.1-$35.0^{\circ}C$, 26.5-96.2% RH) (p<0.05); however, there was no significant correlation between the two parameters. The ratio of calcium activity to total calcium concentration decreased after August compared with the values in June and July (p<0.05). The calcium activity in fresh milk was positively correlated with milk yield (r = 0.45, p<0.01) and negatively correlated with milk lactose content (r = -0.53, p<0.01). These results suggest that the calcium activity in milk could be affected by ambient temperature and might be associated with milking production in hot season.

• Journal of the Korean Magnetic Resonance Society
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• v.2 no.2
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• pp.131-140
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• 1998

The calcium-induced structural changes in the skeletal muscle regulatory protein troponin C (NTnC) involve a transition from a ‘closed’to an ‘open’structure with the concomitant exposure of a large hydrophobic interaction site for target proteins. Structural studies have served to define this conformational change and elucidate the mechanism of the linkage between calcium binding and the induced structural changes. There are now several structures of NTnC available from both NMR and X-ray crystallography. Comparison of the calcium bound structures reveals differences in the level of opening. We have considered the concept of a flexible open state of NTnC as a possible explanation for this apparent discrepancy. We also present simulations of the closed-to-open transition which are in agreement with the flexibility concept and with experimental energetics data.

• Journal of Korea Technical Association of The Pulp and Paper Industry
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• v.34 no.5
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• pp.18-38
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• 2002

A previous study on the model coatings based on latex-bound plastic pigment coatings (1) has been extended to latex-bound No. 1 clay, ultra-fine ground calcium carbonate (UFGCC), and clay-carbonate pigment mixture coatings, which are being widely used in the paper industry. The latex binder used was a good film-forming, monodisperse S/B latex or 0.15$\mu\textrm{m}$. No. 1 clay was representative of plate-like pigment particles, whereas UFGCC was of somewhat rounded rhombohedral pigment particlel. Both of them had negatively skewed triangular particle size distributions having the mean particle suet of 0.7${\mu}{\textrm}{m}$ and 0.6$\mu\textrm{m}$, respectively. Their packing volumes were found to be 62.5% and 657%, respectively. while their critical pigment volume concentrations (CPVC's) were determined to be 52.7% and 50.5% ( average of 45% caused by the incompatibility and 55.9% extrapolated) by coating porosity, respectively. Each pigment/latex coating system has shown its unique relationship between coating properties and pigment concentrations, especially above its CPVC. Notably, the clay/latex coating system hat shown higher coating porosity than the UFGCC/latex system at high pigment concentrations above their respective CPVC's. It was also found that their coating porosity and gloss were inter-related to each other above the CPVC's, as predicted by the theory. More interestingly, the blends of these two pigments have shown unique rheological and coating properties which may explain why such pigment blends are widely used in the industry. These findings have suggested that the unique structure of clay coatings and the unique high-shear rheology of ground calcium carbonate coatings can be judiciously combined to achieve superior coatings. Importantly, the low-shear viscosity of the blends was indicative of their unique packing and coating structure, whereas their high-shear rheology was represented by a common mixing rule, i.e., a viscosity-averaging. Transmission and scanning electron and atomic force microscopes were used to probe the state of pigment / latex dispersions, coating surfaces, freeze fractured coating cross-sections, and coating surface topography. These microscopic studies complemented the above observations. In addition, the ratio, R, of CPVC/(Pigment Packing Volume) has been proposed as a measure of the binder efficiency for a given pigment or pigment mixtures or as a measure of binder-pigment interactions. Also, a mathematical model has been proposed to estimate the packing volumes of clay and ground calcium carbonate pigments with their respective particle size distributions. As well known in the particle packing, the narrower the particle size distributions, the lower the packing volumes and the greater the coating porosity, regardless of particle shapes.

• Journal of Photoscience
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• v.9 no.2
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• pp.281-283
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• 2002

S-modulin in frog or its bovine homologue, recoverin, is a 26 kDa EF-hand $Ca^{2+}$-binding protein found in rod photoreceptors. The $Ca^{2+}$ -bound form of S-modulin binds to rhodopsin kinase (Rk) and inhibits its activity. Through this regulation, S-modulin is believed to modulate the light-sensitivity of a rod. In the present study, we tried to identify the interaction site of the $Ca^{2+}$ -bound form of S-modulin to Rk. First, we mapped roughly the interaction regions by using partial peptides of S-modulin. The result suggested that a specific region near the amino terminus is the interaction site of S- modulin. We then identified the essential amino acid residues in this region by using S-modulin mutant proteins: four amino acid residues were suggested to interact with Rk. These residues are located in a small closed pocket in the $Ca^{2+}$-free, inactive form of S-modulin, but exposed to the surface of the molecules in the $Ca^{2+}$ -bound, active form of S-modulin. Two additional amino acid residues were found to be crucial for the $Ca^{2+}$ -dependent conformational changes of S-modulin. The present study firstly identified the functional site of S-modulin, a member of a neuronal calcium sensor protein family.in family..

• Molecules and Cells
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• v.24 no.2
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• pp.276-282
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• 2007

Protein phosphorylation is one of the major mechanisms by which eukaryotic cells transduce extracellular signals into intracellular responses. Calcium/calmodulin ($Ca^{2+}/CaM$)-dependent protein phosphorylation has been implicated in various cellular processes, yet little is known about $Ca^{2+}/CaM$-dependent protein kinases (CaMKs) in plants. From an Arabidopsis expression library screen using a horseradish peroxidase-conjugated soybean calmodulin isoform (SCaM-1) as a probe, we isolated a full-length cDNA clone that encodes AtCK (Arabidopsis thaliana calcium/calmodulin-dependent protein kinase). The predicted structure of AtCK contains a serine/threonine protein kinase catalytic domain followed by a putative calmodulin-binding domain and a putative $Ca^{2+}$-binding domain. Recombinant AtCK was expressed in E. coli and bound to calmodulin in a $Ca^{2+}$-dependent manner. The ability of CaM to bind to AtCK was confirmed by gel mobility shift and competition assays. AtCK exhibited its highest levels of autophosphorylation in the presence of 3 mM $Mn^{2+}$. The phosphorylation of myelin basic protein (MBP) by AtCK was enhanced when AtCK was under the control of calcium-bound CaM, as previously observed for other $Ca^{2+}/CaM$-dependent protein kinases. In contrast to maize and tobacco CCaMKs (calcium and $Ca^{2+}/CaM$-dependent protein kinase), increasing the concentration of calmodulin to more than $3{\mu}M$ suppressed the phosphorylation activity of AtCK. Taken together our results indicate that AtCK is a novel Arabidopsis $Ca^{2+}/CaM$-dependent protein kinase which is presumably involved in CaM-mediated signaling.