• Korean Journal of Veterinary Research
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• v.29 no.2
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• pp.69-73
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• 1989

Nine strains of Yersinia species isolated from pigs and dogs in Korea, comprising 5 strains of Yersinia enterocolitica, 2 strains of Y kristensenii and each strain of Y pseudotuberculosis and Y intermedia, were examined for the presence of virulence-associated plasmids, calcium dependency and provocation of guinea pig conjuntivitis($Ser{\acute{e}}ny$ test). Three strains of Y enterocolitica isolated from pigs were positive in calcium dependency and harbored one plasmid of about 45 megadalton, but negative in $Ser{\acute{e}}ny$ test.

• Development and Reproduction
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• v.7 no.2
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• pp.95-103
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• 2003

Intracellular calcium is an important physiological factor in most cells, and ruthenium red and ryanodine play an important role as calcium modulators. Ruthenium red inhibits calcium-induced calcium release(CICR) from the intracellular calcium store. Ryanodine activates calcium release through ryanodine channel. The present experiment was performed to investigate the effects of two modulators on calcium ion metabolism and to determine their dose-dependency in oocyte and early embryo of mouse. Intracellular calcium ion concentration was measured in realtime by using confocal laser scanning microscope(CLSM) after loading of Fluo-3/AM in mouse oocytes and early embryos. Ruthenium red decreased intracellular calcium ion concentration in oocytes and early embryos at its high concentration(30, 300 $\mu$M). Ryanodine increased intracellular calcium ion concentration in oocytes and early embryos in low concentration(0.01 $\mu$M) but decreased that at higher concentrations(1, 10 $\mu$M). These results indicate that two modulators affected calcium ion metabolism in oocyte and early embryo of mouse, and their dose-dependency was different from somatic cell including myocytes.

• Korean Journal of Microbiology
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• v.26 no.3
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• pp.262-269
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• 1988

In order to investigate the virulence of Y. enterocolitica isolated in Korea, all necessary experiments were done including several virulence determinating tests-autoagglutination test, calcium-dependency test, HeLa cell invasion test, Sereny test, crystal violet binding test, and electrophoresis for plasmid pattern. The obtained results are as follows: The virulent strains of Y. enterocolitica revealed positive reactions on autoagglutination test, cacium-dependency test, and drystal violet binding test, while the avirulent strains did not. A positive reaction was observed only at $37^{\circ}C$ implying that the expression of virulence is temperature-dependent. In Sereny test, the standard reference virulent strain (serotype 0:8) showed positive reactions while the virylent experimental strains (serotype 1:3, 0:9) revealed negative results, which indicates that the virulence of Y. enterocolitica in experimental animals varied according to their serotypes. Most of the virulent strains contained 36-38Mdal plasmids, but the avirulent strains did not. In addition, it was noted that autoagglutination, calcium-dependency, and crystal violet binding were related to the presence of plasmids.

• Journal of Pharmaceutical Investigation
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• v.38 no.5
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• pp.295-302
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• 2008

In the present study, we examined the changes of uptake and efflux of taurine under various conditions inducing oxidative stress using rat conditionally immortalized syncytiotrophoblast cell line, TR-TBT cell, as blood-placental barrier in vitro model. In addition, we identified the characteristics of taurine transport in TR-TBT cells including general features, besides effect of calcium ion on taurine transport. Taurine uptake showed time, $Na^+$ and $Cl^-$ dependency, and was decreased by PKC activator in TR-TBT cells. Also, calcium free condition decreased taurine uptake and evoked taurine efflux in the cells. Oxidative stress induced the change of taurine transport in TR-TBT cells, but the changes were different depending on the types of stimulation inducing oxidative stress. The taurine uptake was increased by TNF-$\alpha$, LPS and DEM stimulation but decreased by $H_2O_2$ and NO stimulation. Also, the taurine efflux was regulated by TNF-$\alpha$ stimulation. In conclusion, the taurine transport through the blood-placental barrier was regulated in oxidative stress conditions, and these results demonstrated that oxidative stress affected the taurine supplies to fetus and taurine level of fetus.

• Journal of Microbiology and Biotechnology
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• v.18 no.4
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• pp.730-734
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• 2008

Barley ${\alpha}$-amylase genes, amy1 and amy2, were separately cloned into the expression vector of $pPICZ{\alpha}A$ and recombinant Pichia strains were established by homologous recombination. Both AMYs from Pichia shared almost identical hydrolysis patterns on short maltooligosaccharides to result in glucose, maltose, or maltotriose. Against insoluble blue starch, AMY1 showed the highest activity at 0.1-5 mM calcium concentration, whereas 15-20 mM was optimal for AMY2. On the hydrolysis of soluble starch, unexpectedly, there was no significant difference between AMYs with increase of calcium. However, the relative activity on various starch substrates was significantly different between AMYs, which supports that the isozymes are clearly distinguished from each other on the basis of their unique preferences for substrates.

• Korean Journal of Community Nutrition
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• v.9 no.3
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• pp.251-262
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• 2004

The aim of this study was to assess the nutrient intakes from infant formula and supplemental foods of 246 healthy infants fed infant formula, aged from 5 to 18 months. Subjects were devided into two groups depending on supplemental food type for weaning, Domestic supplemental foods (mainly home-made, n = 129) and Delivery supplemental foods (mainly commercially-delivered, n = 117). Four subgroups were assigned to 5-6 months, 7-8 months, 9-11 months, and 12-18 months by ages, respectively. Dietary assessment was carried out using 24-hour-recall method. Formula intakes in the delivery group tended to decrease accordingly with the ages. However, in the domestic group, formula intakes up to 8 months were similar and decreased after 9 month. Energy, protein, calcium and iron intakes from infant formula and supplemental foods were assessed. Energy intake at 12-18 months were lower than the RDA in both groups. Daily intake of protein and calcium at all ages were much higher than the RDA in both groups. Therefore, protein and calcium overnutrition were elucidated. Especially, protein intake at 5-6 months, calcium intake at all ages from infant formula was higher than the RDA in both groups. Iron intake at 5-6 months from infant formula were higher than the RDA. Consequently, as for infant formula, it was suggested that not only formula intakes but also nutrient content in formula should be reconsidered. On the other hand, nutrient intakes from supplemental foods in the domestic group tended to be higher than that of the delivery group. Especially at 9-11 months, significant differences between the two groups were observed. This may be due to high dependency on commercial powdered baby food in the domestic group. This study revealed that daily nutrient intakes of formula-fed infants are desirable but nutrient intakes from infant formula are too high. Conclusively, this study suggests that as the age of infants increases, formula intakes should be controlled and various supplemental foods besides commercially powdered baby food should be appropriately provided.

• Journal of Applied Biological Chemistry
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• v.42 no.1
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• pp.19-24
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• 1999

Plants have been shown to contains $Ca^{2+}$/calmodulin-stimulated GAD and NAD kinase. To test how calmodulin and calmodulin methylation affect the activation of GAD and NAD kinase, GAD and NAD kinase were partially purified from tobacco plants. GAD was also partially purified from E. coli transformed with a plasmid carrying a cloned tobacco GAD gene. We find that GAD from the transformed E. coli showed 60-fold $Ca^{2+}$/calmodulin-dependent activation. However, GAD from tobacco plants was stimulated only about 3.8-fold by the addition of calmodulin in the presence of calcium, suggesting high background activity of the enzyme was possibly due to bound endogenous tobacco calmodulin. There were no significant differences in the tobacco GAD activator properties between calmodulins. A monoclonal antibody against petunia GAD interacted strongly with both GAD from tobacco plants and GAD from cloned gene. NAD kinase from tobacco plants showed a complete $Ca^{2+}$/calmodulin dependency for activity. Unmethylated calmodulins activated GAD in a manner similar to methylated calmodulin. However, the maximum level of NAD kinase activation obtained with unmethylated calmodulins is approximately 4-fold higher than methylated calmodutins. These data suggested that endogenous tobacco calmodulin may interact more tightly with GAD than NAD kinase and that calmodulin methylation affects the activator properties of calmodulins for tobacco NAD kinase but not for GAD.

• Proceedings of the Korean Powder Metallurgy Institute Conference
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• 2006.09b
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• pp.1082-1083
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• 2006

Carbon nanotube (CNT) reinforced hydroxyapatite (HAp) composites were fabricated by using the spark plasma sintering process with surfactant modified CNT and HAp nano powder. Without the dependency on sintering temperature, the main crystal phase existed with the HAp phase although a few contents of ${\beta}-TCP$ (Tri calcium phosphate) phase were detected. The maximum fracture toughness, $(1.27\;MPa.m^{1/2})$ was obtained in the sample sintered at $1100^{\circ}C$ and on the fracture surface a typical intergranular fracture mode, as well as the pull-out pmhenomenon of CNT, was observed.

• Korean Journal of Microbiology
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• v.46 no.1
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• pp.80-85
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• 2010

Two different ${\alpha}$-amylase isozymes (AMY1 and AMY2) found in barley malt share up to 80% of amino acid sequence identity with each other, but their enzymatic properties differ remarkably. AMY1 shows the highest activity at low concentration of calcium ion, while AMY2 is highly active at high calcium concentration. Meanwhile, BASI (Barley ${\alpha}$-Amylase/Subtilisin Inhibitor) protein specifically inhibits only AMY2. In the present study, three separate regions in AMY genes (I, II, and III) were assigned on the basis of restriction enzyme sites and four kinds of chimeric amylases have been obtained by swapping a part of regions with each other. Each chimera gene was successfully over-expressed in Pichia pastoris. From the results of enzymatic characterization, both AMY211 and AMY122 showed the mixed or intermediate type of calcium-dependent activity between AMY1 and 2. Meanwhile, only AMY221 chimera could be significantly inhibited by BASI protein. As a result, it can be proposed that some amino acid residues in the region I and II, except region III, of barley ${\alpha}$-amylases play very important roles in calcium-dependency and interaction with BASI.

• Korean Journal of Veterinary Research
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• v.32 no.2
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• pp.181-194
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• 1992

Two hundred and eighty nine strains of Yersinia enterocolitica isolated from healthy pigs were tested for the presence of 40~50 Megadalton virulence-associated plasmids and plasmidmediated in vitro virulence-associated properties, i.e., congo red uptake, calcium dependency, autoagglutination, CRMOX reaction, crystal violet binding and pyrazinamidase reaction. The correlationships between in vitro virulence-associated properties and the presence of 220 Kdalton outer membrane protein(OMP) were examined in strains with or without virulence-associated plasmids. The correlationships between the presence of plasmids on the production of the OMP and the expression of in vitro virulence-associated properties were studied with $CRMOX^+$ strains and acridine orangecured $CRMOX^-$ mutants. The results were as follows : 1. Of the in vitro virulence-associated tests with 289 strains of Y enterocolitica, 275 strains (95.2%) were positive for pyrazinamidase test, and followed by in order of crystal violet binding test, 226 (79.2% ) ; CRMOX test, 190 (65.7%) ; autoagglutination test, 1.85(64.0%) : calcium dependency test, 86 (29.8%) and congo red uptake test, 47(16.3%). 2. The correlationship between autoagglutination and CRMOX test(r=0.90) was highly significant (p<0.01). 3. In 190 strains(65.7%) bearing the virulence-associated plasmids(MW 40~50 Mdalton), the correlation between the presence of plasmids and their in vitro virulence-associated properties were highest with CRMOX test(r=0.93) and followed by in orders of AAG test(0.81), CV test(0.46), PYZ test(0.37) and CD test(0.18), but no correlationship between the presence of plasmids and CR test(-0.11). 4. The $CRMOX^+$ strains produced the 220 Kdalton OMP when they were cultured at $37^{\circ}C$, but not at $26^{\circ}C$. The presence of 220 Kdalton OMP was correlated significantly with in vitro virulence properties and the presence of virulence-associated plasmid, respectively. 5. In the isogenic $CRMOX^-$ mutant strains, of which plasmid were cured by treatment with acridine orange not only in vitro virulence-associated properties(CR 100%, CD 100%, AAG 82.6%, CV 58.3%) disappeared but also 220 Kdalton OMP(100%) was not produced. These results indicate that the positive CRMOX reaction is plasmid-mediated and the CRMOX test is potential as an in vitro virulence tests with Y enterocolitica.