• Korean Journal of Microbiology
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• v.48 no.2
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• pp.109-115
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• 2012

Helicobacter pylori is an important factor of chronic gastritis, digestive ulcer, and stomach cancer. CagL, a virulence factor of H. pylori, is well-known as a pilus protein which acts as adhesion to host cell and a component of Type 4 secretion system. In this study, we evaluated the protective response of recombinant CagL protein (rCagL) using Mongolian gerbil animal model for H. pylori infection. The cagL gene was cloned from 26695 H. pylori followed by over-expression and purification of the protein in E. coli. Mongolian gerbils were immunized with rCagL protein mixed with aluminum adjuvant via intramuscular injections once a week during 4 weeks. At a week after the last immunization, the Mongolian gerbils were administrated with H. pylori 7.13 strain into the stomach and sacrificed to measure antibody titer on rCagL by ELISA and bacterial colonization in the stomach, and to examine the histopathological changes and cytokine expression at 6 week after challenge. Antibody titers on recombinant protein were significantly increased from a week after the first immunization. There was no significant change of the number of bacterial colony between control group and immunized group. The relative stomach weight was significantly decreased in immunized group, but the significant change of histopathological assessment was not observed in the stomach. Cytokine expression such as IL-$1{\beta}$ and KC also was not significantly different between control and immunized groups. These results indicate that rCagL could effectively induce the formation of the specific IgG antibodies. However, bacterial colonization and histopathological lesions could not be inhibited by the immunization in the stomach, indicating not enough protection against H. pylori infection. We consider that along with CagL other adequate antigens could be needed stimulating immune response and inducing protective effects against gastric disease, and also a better adjuvant could be considered.

• Journal of the korean academy of Pediatric Dentistry
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• v.30 no.2
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• pp.217-228
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• 2003

Co-ordinate growth of the brain and skull is achieved through a series of tissue interactions between the developing brain, the growing bones of the skull and the sutures that unite the bones. Craniosynostosis, the premature fusion of cranial sutures, presumably involves disturbance of these interactions. Bmp2, one of bone morphogenetic proteins (Bmps), is involved in the regulation of the shapes of individual bones and the relative proportions of the skeleton. Mutations in the homeobox gene Msx2, known as a downstream gene of Bmp, cause Boston-type human craniosynostosis. The phenotype of Dlx5 homozygote mutant mouse presents craniofacial abnormalities including a delayed ossification of calvarial bone. These facts suggest important roles of Bmp2, Msx2 and Dlx5 genes in the cranial bone growth and suture morphogenesis. To elucidate the function of these molecules in the early morphogenesis of mouse cranial sutures, we first analyzed by in situ hybridization the expression of Bmp2(E15-18), Msx2 and Dlx5 genes in the developing sagittal suture of calvaria during the embryonic stage. Bmp2 mRNA was intensely expressed in the osteogenic fronts and also at the low level in the periosteum of parietal bones during embryonic stage, Msx2 mRNA was intensely expressed in the sutural mesenchyme and mildly expressed in the dura mater during the embryonic stage. Dlx5 mRNA was intensely expressed osteogenic fronts and parietal bones. To further examine the role of Bmp signaling in cranial suture, we did in vitro experiments in E15.5 mouse calvarial explants. Interestingly, implantation of Bmp2-soaked beads onto the osteogenic fronts after 48 hours organ culture resulted in the increase of the tissue thickness and cell number around Bmp2 beads, compared to BSA control beads. In addition Bmp2 induced etopic expressions of Msx2 and Dlx5 genes. On the other hand, overexpression of FGF2 did not induce the expression of Msx2 and Dlx5. Taken together, these data indicate that Bmp2 signaling molecule has a important role in regulating the cranial bone growth and early morphogenesis of cranial suture. We also suggest that Bmp signaling is involved in all the stages of osteogenesis of cranial bones and the maintenance of cranial suture by regulating Msx2 and Dlx5 genes, and that Msx2 and Dlx5 genes are specific transcription factors of Bmp signaling pathway.

• Journal of the Korean Society of Food Science and Nutrition
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• v.35 no.3
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• pp.366-372
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• 2006

This study is to investigate the physicochemical properties of differently pretreated chestnut starches during starch isolation and to examine their gelatinization properties by both heat and alkali treatments. One kind is starch A made by alkali method from peeled chestnut. The other is starch B made from chestnut with the outer layer. The results are as follows. Starch A has higher water binding capacity of 86.9% than starch B with 80.66%. Swelling powers of both starch A and B increased rapidly from $60^{\circ}C\;to\;80^{\circ}C$ in both, and since then it has changed a bit. Both began to show their solubility at $60^{\circ}C$ and increased continuously as the temperature went up. Starch A has higher swelling power and solubility than starch B. In iodine reaction, starch A has higher ${\lambda}max$ and absorbance at ${\lambda}max$ than starch B. X-ray diffraction patterns showed that starch A is type $C_b$ and that starch B is type B. Starch B has higher relative crystallinity of 37.0% than starch A with 36.2%. The results by differential scanning calorimetry revealed that starch A gelatinized from $66.95^{\circ}C$ to $77.5^{\circ}C$ and its enthalpy is 2.04 cal/g. And starch B gelatinized from $67.09^{\circ}C\;to\;77.5^{\circ}C$, and its enthalpy is 2.29 cal/g. Amylograms of chestnut starch at 6.5% concentration indicated that starch B needs higher onset temperature when beginning to gelatinize than starch A does. But starch A shows much higher peak viscosity, breakdown and setback than starch B does. Starch A shows higher viscosity, gel volume, and optical transmittance in gelatinization properties by alkali than starch B does.

• Economic and Environmental Geology
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• v.41 no.6
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• pp.695-708
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• 2008

In order to evaluate the geochemical behaviors of elements with waste rocks in the abandoned Jangpoong Cu mine area, total concentration analysis and leaching experiments were performed. The content of elements within waste rocks compared with background values decreased in order of As>>Cu>Pb>Cd>Co. Leaching experiments were carried out at various extraction environments, considering the acid rain ($0.00001{\sim}0.001N\;HNO_3$) and the acid mine drainage ($0.001{\sim}0.1N$ HNO3). After 24 hours of reaction with different acidic solution, the leaching characteristics of waste rocks were classified into three types according to final pH of leaching solution. Type I refers to the case that the final pH of leaching solution was lower than that of the reaction solution due to the dissolution of acidic minerals from rocks, while type 2 and 3 refer to the case that the final pH maintained higher than that of the reaction solution. Theses types include in acid buffering minerals such as clay minerals and carbonate minerals. The leaching characteristics of the elements after the reaction could be categorized into As-Co-Fe, Cu-Mn-Cd-Zn, and Pb. As-Co-Fe started to get leached under 2.5 of pH regardless of changes in the final pH, and Cu-Mn-Cd-Zn showed different initial leaching pH according to the types of final pH changes. Based on the pH value where leaching started regardless of leaching concentration, the relative mobility of each element was in the order of Mn Zn>Cd>Cu>>Fe Co>As>Pb. Thus, more higher mobility elements(Zn, Mn and Cu) were leached by reacting with acid rain water. Acid mine drainage may result in distributions of elements having relatively less mobility(As, Fe, Co and Pb).

• Applied Microscopy
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• v.29 no.3
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• pp.383-403
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• 1999

This study was performed to investigate the immunohistochemical and ultrastructural characterization of the choline acetyltransferase (ChAT)-immunoreactive nerve cells in the diagonal band of Broca of the rat basal forebrains, utilizing techniques of immunohistochemical and immunocytochemical microscopy. The ChAT-immunoreactivities were shown within neuronal cell bodies and processes by the light micoscope. According to cell shape and ratio of long axis vs short axis of cell body, the ChAT-immunoreaclive nerve cells in both vertical and horizontal limbs of the diagonal band of Broca were classified into 6 types. at the light microscopic level; round, oval, elongated, fusiform, triangular and polygonal types. As a result of the electron microscopic observation, the ChAT-immunoreactivated products appeared on the outer nuclear envelope, membranes of rough endoplasmic reticula (rER), free ribosomes and polysomes. Each cell type was subdivided into subtype I and II according to the several criteria such as volume of cell body, nuclear size relative to the cytoplasm, kinds and distribution of cell organelles and numbers and sorts of synapses. The subtype I of immnunoreactive nerve cells had large cell body and a small nucleus showing shallow indentations of nuclear evelope. In this subtype I with abundant cytoplasm, rER were well differentiated. Their long cisternae were parallelly ditributed and lamellated. One or two lamellar bodies and nematosomes were observed. The subtype II cell had small cell body and a large nucleus with deep indentations of nuclear envelope. In this subtype II with small cytoplasm, the rER were irregularly distributed and the lamellar body and nematosome were not found. A few axosomatic synapses in the subtype I and II were shown to be symmetric or asymmetric. The ratios of the symmetric synapse to the asymmetric one were investigated to be 1 : 2 and 1 : 4 in the subtype I and II, respectively. The axodendritic ones were almost asymmetric. But, the fusiform and triangular immunoreactive nerve cells were shown only to be subtype I. According to observations in this study, it is considered that the ultrastructural characterization in the 2 subtypes of each cell type may reflect the differences of the metabolic activities and projecting distances to the target cells.

• Journal of Life Science
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• v.23 no.1
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• pp.15-23
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• 2013

AprE51 from Bacillus amyloliquefaciens CH51 is a 27 kDa subtilisin-like protease with fibrinolytic activity. AprE51-6 showing increased catalytic activity was produced previously. To enhance the thermostability of AprE51-6, 2 residues, Gly-166 and Asn-218 based on B. subtilis subtilisin E were mutated by site-directed mutagenesis. The results of the mutational analysis showed that substitution of arginine for Gly-166 (AprE51-7) increased the fibrinolytic activity 1.8-fold. An N218S mutant (AprE51-8) also increased the fibrinolytic activity up to 4.5-fold in a fibrin plate assay. Purified AprE51-7 and AprE51-8 mutants had a 1.9- and a 2.5-fold higher $k_{cat}$, respectively, and a 2.1-1.9-fold lower $K_m$, respectively. This resulted in a 3.8- and a 4.7-fold increase in catalytic efficiency ($k_{cat}/K_m$), respectively, relative to that of wild-type AprE51. AprE51-8 had a broader pH range than AprE51-6 and nattokinase, especially at an alkaline pH value. In addition, AprE51-8 showed higher thermostability than AprE51-6 at $60^{\circ}C$. The half-lives of AprE51-7 and AprE51-8 at $50^{\circ}C$ were 21.5 and 27.3 min, respectively, which are 2.0 and 2.6 times longer, respectively, than that of the wild-type AprE51.

• Korean journal of applied entomology
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• v.38 no.2
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• pp.101-107
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• 1999

Queens of Korean native bumblebee species, Bombus ignitus were collected from the field in the spring of 1998 to investigate their ecological characteristics and reared under the laboratory conditions; 3 0 f 1$^{\circ}$C, 55 25% of relative humidity and darkness. As a result, 94.4% of the queens laid eggs in 6.00f 2.41 days after the field collection. The first worker and the first drone from the egg cell of the collected queens appeared in 18.90 f 1.16 and 68.96 f 3.94 days, respectively. In the colony foundation, 86.1% of the collected queens founded their colony, and 96.8% of them produced new queens in 71.14f 6.88 days. The life span of the colony-founded queen covered for 86.90f24.35 days, whereas that of non-colonized queen for 15.67 f 7.37 days. One colony sized as big as 987.82f 145.84 in number, composing of workers (188.79f 18.23), drones (691.10f 159.24) and queens (107.93 f47.48). Copulation lasting time of queen and drone took 23.00f 7.33 minutes in average. In a new queen emergence, there were two patterns; single peak type of new queen emergence in 12.31 k6.95 days and two peaks type of it in 74.00f 17.61 days. The number of eggs per egg cell of new queen averaged as 8.86f 4.00 grains and an egg period covered 3 days after oviposition. An egg cell sized as 7.15 f0.88 mm in a width, 7.79 f 1 .l 1 mm in a length and 3.33 f 0.23 mm in a height. An egg shaped as banana and sized as 1.22 f 0.09 mm in a width and as 3.5 1 f 0.21 mm in a length, weighing as 2.70 + 0.30 mg.

• The Journal of the Petrological Society of Korea
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• v.24 no.3
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• pp.165-180
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• 2015

Jurassic granite from Geochang was analysed with respect to the characteristics of the rock cleavage. The phases of distribution of microcracks were well evidenced from the enlarged photomicrographs(${\times}6.7$) of the thin section. In this study, the length - cumulative frequency diagrams were used for expressing the distribution characteristics of microcrack. The diagrams for the six directions were arranged in the magnitude of density(${\rho}$). These diagrams show an order of H2 < H1 < G2 < G1 < R2 < R1 from the related chart. Among six diagrams, the diagram for hardway 2(H2) occupies the lowermost region on the left. On the contrary, the diagram for rift 1(R1) occupies the uppermost region on the right. Curve patterns of the two diagrams change from uniform to exponential distribution type in accordance with the increased density. The overall distribution characteristics of the diagrams were well evidenced from the magnitude of the exponent(${\lambda}$) and length of line oa related to the exponential straight line. The magnitude of exponent governing the values of slope(${\theta}$) is inversely proportional to the values of microcrack parameters such as number(N), length(L) and density. On the contrary, length of line oa is directly proportional to the values of the above three parameters. Above microcrack parameters related to the order of arrangement of diagrams show an order of hardway(H1 + H2) < grain(G1 + G2) < rift(R1 + R2). The distribution characteristics of progressive variation are found among the six diagrams. The order of arrangement of the diagrams indicates a relative magnitude of the rock cleavage. Meanwhile, the parameters such as slope, exponent, density and length of line oa were arranged in an order of H2 < H1 < G2 < G1 < R2 < R1. The variation curves of a smooth quadratic function are shown from the related chart. From the correlation chart between density and the above parameters, a common regularity following power-law correlation function was derived. Finally, the analysis for the rock cleavage was conducted through the combination between the diagram and microcrack parameter. This type of combination contribute to the progressivity in evaluation for the rock cleavage.

• The Journal of the Petrological Society of Korea
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• v.25 no.1
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• pp.13-27
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• 2016

The characteristics of the rock cleavage inherent in Jurassic granite from Geochang were analysed. The phases of distribution of microcrack spacings were derived from the enlarged photomicrographs(${\times}6.7$) of the thin section. The evaluation for the six directions of rock cleavages was performed using nine parameters such as (1) frequency of microcrack spacing(N), (2) frequency ratio(${\leq}1mm$ and 4 mm >) to total spacing frequency(N:191), (3) spacing ratio(${\leq}1mm$) to total spacing(118.49 mm), (4) mean spacing($S_{mean}$), (5) difference value($S_{mean}-S_{median}$) between mean spacing and median spacing($S_{median}$), (6) density of spacing, (7) median spacing, (8) reduction ratio of spacing frequency to length frequency and (9) magnitude of exponent(${\lambda}$ and b) related to the distribution type of diagram. Especially the close dependence between the above spacing parameters and the parameters from the spacing-cumulative frequency diagrams was derived. The results of correlation analysis between the values of parameters for three rock cleavages and those for three planes are as follows. The values of (I) parameters(1, 2 and 3), (II) parameters(4, 5 and 6), (III) parameter(7), (IV) parameter(8) and (V) parameter(9) show the various orders of H(hardway, H1+H2) < G(grain, G1+G2) < R(rift, R1+R2), R < G < H, R < H < G, G < H < R and H < G < R, respectively. On the contrary, the values of the above four groups(I~IV) of parameters for three planes show reverse orders. This type of correlation analysis is useful for discriminating three quarrying planes. Six spacing-cumulative frequency diagrams were arranged in increasing order on the value of main parameter($S_{mean}-S_{median}$). These diagrams show an order of R2 < R1 < G2 < G1 < H2 < H1 from the related chart. In other words, the above six diagrams can be summarized in order of rift(R1+R2) < grain(G1+G2) < hardway(H1+H2). These results indicate a relative magnitude of rock cleavage related to microcrack spacing. Especially, the above main parameter could provide advanced information for prediction the order of arrangement among the diagrams.

• 한국균학회소식:학술대회논문집
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• 2014.10a
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• pp.9-9
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• 2014

Null mutants generated by targeted gene replacement are frequently used to reveal function of the genes in fungi. However, targeted gene deletions may be difficult to obtain or it may not be applicable, such as in the case of redundant or lethal genes. Constitutive expression system could be an alternative to avoid these difficulties and to provide new platform in fungal functional genomics research. Here we developed a novel platform for functional analysis genes in Magnaporthe oryzae by constitutive expression under a strong promoter. Employing a binary vector (pGOF1), carrying $EF1{\beta}$ promoter, we generated a total of 4,432 transformants by Agrobacterium tumefaciens-mediated transformation. We have analyzed a subset of 54 transformants that have the vector inserted in the promoter region of individual genes, at distances ranging from 44 to 1,479 bp. These transformants showed increased transcript levels of the genes that are found immediately adjacent to the vector, compared to those of wild type. Ten transformants showed higher levels of expression relative to the wild type not only in mycelial stage but also during infection-related development. Two transformants that T-DNA was inserted in the promotor regions of putative lethal genes, MoRPT4 and MoDBP5, showed decreased conidiation and pathogenicity, respectively. We also characterized two transformants that T-DNA was inserted in functionally redundant genes encoding alpha-glucosidase and alpha-mannosidase. These transformants also showed decreased mycelial growth and pathogenicity, implying successful application of this platform in functional analysis of the genes. Our data also demonstrated that comparative phenotypic analysis under over-expression and suppression of gene expression could prove a highly efficient system for functional analysis of the genes. Our over-expressed transformants library would be a valuable resource for functional characterization of the redundant or lethal genes in M. oryzae and this system may be applicable in other fungi.