• Journal of Plant Biology
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• 제39권3호
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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.

• 한국임상수의학회지
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• 제9권2호
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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.

• 대한방사선기술학회지:방사선기술과학
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• 제32권1호
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• pp.69-78
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• 2009

연구배경 및 목적 : 심혈관 질환은 서구에서는 이미 오래전부터 가장 흔한 사망 원인으로 알려져 있고, 우리나라를 포함한 동양에서는 최근 식생활 습관의 변화와 함께 급격히 증가하는 추세에 있다. 특히 허혈성 심장질환은 통계청 curve : 곡선 $1996{\sim}2006$을 보면 향후 $5{\sim}10$년 사이 허혈성 심장질환이 1위로 올라 갈 것으로 전망할 수 있다. 이에 보다 정확하고 정밀한 선별검사가 필요함을 인식할 수 있겠다. 목적은 Calcium-scoring의 총 정량화수치에 영향을 주는 위험인자와의 연관성을 알아보고, Calciumscoring이 선별검사로의 타당성을 검증하며, 국민보건에 이바지 하고자 한다. 연구방법 및 결과 : 인천광역시에 위치한 G병원으로 2008년 2월 1일부터 6월 30일까지 가슴 흉통으로 내원한 환자 중 Calcium-scoring과 Coronary CT angiography를 동시에 시행한 환자 중 무작위 표본추출로 70명을 선정 하였으며, Calcium-scoring t value에 영향을 주는 것으로 알려진 위험인자를 각각의 연관성을 분석한 결과 고지혈증, 고혈압이 유의수준 $\alpha$ < 0.05하에서 Calcium-scoring t value를 높임을 알 수 있었고, Calcium-scoring t value를 Coronary CT angiography 결과와 비교분석 한 결과 관상동맥 석회화 수치는 관상동맥 내의 동맥경화와의 총량과 상관관계가 높다는 결과를 얻을 수 있었고, Coronary CT angiography result를 확진검사로 ROC curve를 분석한 결과 Area의 크기가 0.833으로 우수한 검사법이라 할 수 있고, Asymptotic 95% Confidence interval Low Bound와 Upper Bound가 모두 0.5 이상이며, Asymptotic Sig.(p)가 0.05보다 작으므로 유용한 검사법임을 알 수 있었다. 결 론 : Calcium-scoring검사가 허혈성 심혈관 질환의 다른 선별검사인 심전도와 운동부하검사 등과 더불어 선별검사로서 손색이 없으며, 운동부하검사의 제한점인 관절 질환을 앓고 있거나, 고령 등의 원인으로 운동이 어려운 경우에는 시행하기 어려운 점을 해결할 수 있는 우수한 선별검사라고 할 수 있겠다.

• Journal of Microbiology and Biotechnology
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• 제14권3호
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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.

• 한국환경농학회지
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• 제16권3호
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• pp.239-244
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• 1997

토양중 tightly bound paraquat의 효과적인 추출방법을 모색코자 HCl 및 $H_2O_2$첨가 그리고, keldahl 분해장치를 이용하여 회수율 및 여과시간의 단축을 조사하였으며, ammonium chloride와 calcium chloride의 농도와 진탕 시간을 달리하여 loosely bound paraquat의 유출정도를 조사한 결과는 다음과 같다. 1. 토양 50g에 70ml의 conc. HCl을 첨가하여 시간별로 진탕하고 여기에 진한 황산을 가하여 1시간동안 환류 추출한 결과, 60분간 진탕할때 기존의 추출방법에 비해 A토양은 1.15배 높은 96.7%의 회수율과 4.64배 단축된 14분의 여과 시간을, B토양은 1.09배 높은 84.0%의 회수율과 4.91배 단축된 11분의 여과시간을 나타냈다. 아울러, 이러한 결과는 회수율의 증가는 물론 여과시간의 단축, pH조정과정에서 과다한 NaOH사용을 통한 총용량의 증가 방지, 그리고 전체적인 실험시간의 단축을 꾀할 수 있었다. 2. HCl의 첨가에 의한 진탕과정 없이 $H_2O_2$만을 첨가하는 경우, $H_2O_2$ 30ml첨가하여 30분간의 환류 재추출하는 것이 적정 수준이었는데, 회수율은 증대하였으나 여과시간의 단축은 거의 없었다. 반면, 50ml의 $H_2O_2$ 첨가만으로도 회수율의 증대는 있었으나 실험의 위험성을 고려시 적절하지 못하다고 생각된다. 3. Kjeldahl 분해장치를 이용한 방법으로 회수율의 증대를 나타내는 경향은 있었으나, 반복간의 큰 오차를 나타내었으므로 추출방법으로 적합하지 못하다고 생각된다. 4. Loosely bound paraquat의 추출을 위해 0.01, 0.1, 1, 10M ammonium chloride와 calcium chloride를 이용하여 1.5, 3, 6, 12, 24시간으로 진탕시간을 달리하여 분석한 결과, 시간, 농도, 추출액의 변화에도 불구하고 모든 처리구에서 paraquat은 검출한계 미만의 수준이었다.

• Asian-Australasian Journal of Animal Sciences
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• 제24권10호
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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.

• 한국자기공명학회논문지
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• 제2권2호
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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.

• 펄프종이기술
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• 제34권5호
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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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• 제9권2호
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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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• 제24권2호
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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.