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Immunohistochemical and Ultrastructural Characterization of the Choline Acetyltransferase-immunoreactive Nerve Cells in the Diagonal Band of Broca of the Rat Basal Forebrains (흰쥐의 전뇌 기저부 대각 Broca대에서 Choline Acetyltransferase 면역반응 신경세포에 대한 면역조직화학 및 미세구조)

  • Back, Seung-Keun;Chung, Young-Wha
    • 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.

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Enhancement of the Thermostability of a Fibrinolytic Enzyme from Bacillus amyloliquefaciens CH51 (Bacillus amyloliquefaciens CH51이 생산하는 혈전용해효소의 열안정성 개선)

  • Kim, Jieun;Choi, Kyoung-Hwa;Kim, Jeong Hwan;Song, Young-Sun;Cha, Jaeho
    • 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.

Ecological Characteristics of Bombus ignitus Smith in Korea (한국산 호박벌(Bombus ignitus Smith)의 생태적 특성)

  • ;;;;M. Bilinski
    • 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.

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Evaluation for Rock Cleavage Using Distribution of Microcrack Lengths (미세균열의 길이 분포를 이용한 결의 평가)

  • Park, Deok-Won
    • 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.

Evaluation for Rock Cleavage Using Distribution of Microcrack Spacings (I) (미세균열의 간격 분포를 이용한 결의 평가(I))

  • Park, Deok-Won
    • 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.

Development of System-Wide Functional Analysis Platform for Pathogenicity Genes in Magnaporthe oryzae

  • Park, Sook-Young;Choi, Jaehyuk;Choi, Jaeyoung;Kim, Seongbeom;Jeon, Jongbum;Kwon, Seomun;Lee, Dayoung;Huh, Aram;Shin, Miho;Jung, Kyungyoung;Jeon, Junhyun;Kang, Chang Hyun;Kang, Seogchan;Lee, Yong-Hwan
    • 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.

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Effect of Harvest Time on Yield and Feed Value of Whole Crop Barleys with Different Awn Types (망 특성이 다른 청보리의 수확시기별 수량 및 사료가치)

  • Song, Tae-Hwa;Han, Ouk-Kyu;Yun, Seong-Kun;Park, Tae-Il;Kim, Kee-Jong
    • Journal of The Korean Society of Grassland and Forage Science
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    • v.31 no.4
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    • pp.361-370
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    • 2011
  • This study was conducted to obtain basic a information on growth characteristics, productivity, and feed values of whole crop barleys with different awn types. The field experiments were conducted on land at Iksan, Jeollabuk-do province in Korea for one year. Six barleys with three different awn types, such as Youngyang (rough), Yuyeon (hood), Youho (hood), Iksan449 (hood), Iksan456 (hood), and Iksan448 (awnless), were used in this study. Barleys were harvested on five growth stages after (DAH); 20 days after heading (20 DAH), 25 DAH, 30 DAH, 35 DAH, and 40 DAH. Heading was earlier in Yuyeon (April 22), while was later in Iksan448 (April 29). The maximum dry matter and total digestible nutrients (TDN) yields, 11.8 and 8.1 ton per ha, respectively, was recorded in Ikan456 harvested at 35~40 DAH, followed by Youngyang, Yuyeon, Iksan448, Youho and the lowest in Iksan449. They were increased significantly with advancing crop maturity. Crude protein content was the highest in Iksan448 (10.9%) and the lowest in Youho (9.7%) and that was decreased significantly with late harvest. Total digestible nutrients (TDN) and relative feed value (RFV) were the highest in Iksan448 (73.0%, 183.8, respectively) and the lowest in Iksan456 (68.0%, 135.4, respectively) and they were increased significantly with late harvest. The mean RFV evaluated by awn types was the highest in Iksan448 (183.8) of awnless, the lowest in Youho (149.5) and Iksan456 (135.4), indicated the insufficiency of hooded type barleys as good fodder use. Therefore, in this study suggest that hooded barley should improve forage productivity as well as quality.

Fabrication and Oxygen Permeation Properties of ${La_{1-x}Sr_{x}B_{1-{\gamma}}Fe_{\gamma}O_{3-{\delta}}$(B=Co, Ga) Perovskite-Type Ceramic Membranes (${La_{1-x}Sr_{x}B_{1-{\gamma}}Fe_{\gamma}O_{3-{\delta}}$(B=Co, Ga) 페롭스카이트 세라믹 분리막의 제조 및 산소투과특성)

  • 임경태;조통래;이기성;한인섭;서두원
    • Membrane Journal
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    • v.11 no.4
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    • pp.143-151
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    • 2001
  • We have fabricated mixed-ionic conducting membranes, L $a_{0.6}$S $r_{0.4}$ $Co_{0.2}$F $e_{0.8}$ $O_{3-}$$\delta$/ and L $a_{0.7}$S $r_{0.3}$G $a_{0.6}$F $e_{0.4}$ $O_{3-}$$\delta$/ by the solid state method. Ceramic membranes consisted of perovskite-type structures and exhibited high relative density, >95%. Especially, dense L $a_{0.6}$S $r_{0.4}$Co $O_{3-}$$\delta$/ layer was coated on the L $a_{0.7}$S $r_{0.3}$G $a_{0.6}$F $e_{0.4}$ $O_{3-}$$\delta$/ membranes by using screen printing technique in order to improve oxygen ion flux. We measured oxygen ion flux on uncoated L $a_{0.6}$S $r_{0.4}$ $Co_{0.2}$F $e_{0.8}$ $O_{3-}$$\delta$/, uncoated L $a_{0.7}$S $r_{0.3}$G $a_{0.6}$F $e_{0.4}$ $O_{3-}$$\delta$/, and coated L $a_{0.7}$S $r_{0.3}$G $a_{0.6}$F $e_{0.4}$ $O_{3-}$$\delta$/ membranes. The L $a_{0.6}$S $r_{0.4}$ $Co_{0.2}$F $e_{0.8}$ $O_{3-}$$\delta$/ membranes showed the highest flux, 0.26 mL/min.$\textrm{cm}^2$ at 90$0^{\circ}C$, after steady state had been reached. The oxygen flux of coated L $a_{0.7}$S $r_{0.3}$G $a_{0.6}$F $e_{0.4}$ $O_{3-}$$\delta$/ membranes showed higher value, 0.19 mL/min.$\textrm{cm}^2$ at 95$0^{\circ}C$. This flux was as much as 2 or 3 times higher than those of uncoated L $a_{0.7}$S $r_{0.3}$G $a_{0.6}$F $e_{0.4}$ $O_{3-}$$\delta$/ membranes. 3-$\delta$/ membranes.X> 3-$\delta$/ membranes.membranes.

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The Effect of Rain on Traffic Flows in Urban Freeway Basic Segments (기상조건에 따른 도시고속도로 교통류변화 분석)

  • 최정순;손봉수;최재성
    • Journal of Korean Society of Transportation
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    • v.17 no.1
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    • pp.29-39
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    • 1999
  • An earlier study of the effect of rain found that the capacity of freeway systems was reduced, but did not address the effects of rain on the nature of traffic flows. Indeed, the substantial variation due to the intensity of adverse weather conditions is entirely rational so that its effects must be considered in freeway facility design. However, all of the data in Highway Capacity Manual(HCM) have come from ideal conditions. The primary objective of this study is to investigate the effect of rain on urban freeway traffic flows in Seoul. To do so, the relations between three key traffic variables(flow rates, speed, occupancy), their threshold values between congested and uncontested traffic flow regimes, and speed distribution were investigated. The traffic data from Olympic Expressway in Seoul were obtained from Imagine Detection System (Autoscope) with 30 seconds and 1 minute time periods. The slope of the regression line relating flow to occupancy in the uncongested regime decreases when it is raining. In essence, this result indicates that the average service flow rate (it may be interpreted as a capacity of freeway) is reduced as weather conditions deteriorate. The reduction is in the range between 10 and 20%, which agrees with the range proposed by 1994 US HCM. It is noteworthy that the service flow rates of inner lanes are relatively higher than those of other lanes. The average speed is also reduced in rainy day, but the flow-speed relationship and the threshold values of speed and occupancy (these are called critical speed and critical occupancy) are not very sensitive to the weather conditions.

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Fragment Analysis for Detection of the FLT3-Internal Tandem Duplication: Comparison with Conventional PCR and Sanger Sequencing (FLT3-ITD 검출을 위한 절편분석법: 일반 중합효소연쇄반응 및 직접염기서열분석법과의 비교)

  • Lee, GunDong;Kim, Jeongeun;Lee, SangYoon;Jang, Woori;Park, Joonhong;Chae, Hyojin;Kim, Myungshin;Kim, Yonggoo
    • Laboratory Medicine Online
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    • v.7 no.1
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    • pp.13-19
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    • 2017
  • Background: We evaluated a sensitive and quantitative method utilizing fragment analysis of the fms-like tyrosine kinase 3-internal tandem duplication (FLT3-ITD), simultaneously measuring mutant allele burden and length, and verified the analytical performance. Methods: The number and allelic burden of FLT3-ITD mutations was determined by fragment analysis. Serial mixtures of mutant and wild-type plasmid DNA were used to calculate the limit of detection of fragment analysis, conventional PCR, and Sanger sequencing. Specificity was evaluated using DNA samples derived from 50 normal donors. Results of fragment analysis were compared to those of conventional PCR, using 481 AML specimens. Results: Defined mixtures were consistently and accurately identified by fragment analysis at a 5% relative concentration of mutant to wild-type, and at 10% and 20% ratios by conventional PCR and direct sequencing, respectively. No false positivity was identified. Among 481 AML specimens, 40.1% (193/481) had FLT3-ITD mutations. The mutant allele burden (1.7-94.1%; median, 28.2%) and repeated length of the mutation (14-153 bp; median, 49 bp) were variable. The concordance rate between fragment analysis and conventional PCR was 97.7% (470/481). Fragment analysis was more sensitive than conventional PCR and detected 11 additional cases: seven had mutations below 10%, three cases represented conventional PCR failure, and one case showed false negativity because of short ITD length (14 bp). Conclusions: The new fragment analysis method proved to be sensitive and reliable for the detection and monitoring of FLT3-ITD in patients with AML. This could be used to simultaneously assess ITD mutant allele burden and length.