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Distribution and Identification of Halophilic Bacteria in Solar Salts Produced during Entire Manufacturing Process (천일염 생산공정별 미생물 분포 조사 및 호염미생물 동정)

  • Na, Jong-Min;Kang, Min-Seung;Kim, Jin-Hyo;Jin, Yong-Xie;Je, Jeong-Hwan;Kim, Jung-Bong;Cho, Young-Sook;Kim, Jae-Hyun;Kim, So-Young
    • Microbiology and Biotechnology Letters
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    • v.39 no.2
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    • pp.133-139
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    • 2011
  • In this study, we determined the changes in microbial numbers in solar salts according to the manufacturing process and storage duration. The salt samples were harvested from salt farms in Shinan (area 2) and Yeonggwang (area 1). They were serially diluted ten-fold and then placed on 4 kinds of cultivable media (mannitol salt agar, eosin methylene blue, plate count agar, and trypticase soy agar). After incubation, we obtained 62 halophilic isolates from the salt samples. Coliform and general bacteria were not detected in all salt samples. By 16S rRNA sequencing analysis, we found 12 kinds of halophilic bacteria belonging to the genera Halobacillus, Halomonas, Bacillus, Idiomarina, Marinobacter, Pseudoalteromonas, Vibrio, Salinivibrio, Virgibacillus, Alteromonas, Staphylococcus and some un-known stains. In our study, we discovered two novel species that have a 16S rDNA sequence similarity below 97%.

Cloning and Characterization of an Endoglucanase Gene from Actinomyces sp. Korean Native Goat 40

  • Kim, Sung Chan;Kang, Seung Ha;Choi, Eun Young;Hong, Yeon Hee;Bok, Jin Duck;Kim, Jae Yeong;Lee, Sang Suk;Choi, Yun Jaie;Choi, In Soon;Cho, Kwang Keun
    • Asian-Australasian Journal of Animal Sciences
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    • v.29 no.1
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    • pp.126-133
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    • 2016
  • A gene from Actinomyces sp. Korean native goat (KNG) 40 that encodes an endo-${\beta}$-1,4-glucanase, EG1, was cloned and expressed in Escherichia coli (E. coli) $DH5{\alpha}$. Recombinant plasmid DNA from a positive clone with a 3.2 kb insert hydrolyzing carboxyl methyl-cellulose (CMC) was designated as pDS3. The entire nucleotide sequence was determined, and an open-reading frame (ORF) was deduced. The ORF encodes a polypeptide of 684 amino acids. The recombinant EG1 produced in E. coli $DH5{\alpha}$ harboring pDS3 was purified in one step using affinity chromatography on crystalline cellulose and characterized. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis/zymogram analysis of the purified enzyme revealed two protein bands of 57.1 and 54.1 kDa. The amino terminal sequences of these two bands matched those of the deduced ones, starting from residue 166 and 208, respectively. Putative signal sequences, a Shine.Dalgarno-type ribosomal binding site, and promoter sequences related to the consensus sequences were deduced. EG1 has a typical tripartite structure of cellulase, a catalytic domain, a serine-rich linker region, and a cellulose-binding domain. The optimal temperature for the activity of the purified enzyme was $55^{\circ}C$, but it retained over 90% of maximum activity in a broad temperature range ($40^{\circ}C$ to $60^{\circ}C$). The optimal pH for the enzyme activity was 6.0. Kinetic parameters, $K_m$ and $V_{max}$ of rEG1 were 0.39% CMC and 143 U/mg, respectively.

Cortical Thickness Estimation Using DIR Imaging with GRAPPA Factor 2 (DIR 영상을 이용한 피질두께 측정: GRAPPA 인자 2를 이용한 비교)

  • Choi, Na-Rae;Nam, Yoon-Ho;Kim, Dong-Hyun
    • Investigative Magnetic Resonance Imaging
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    • v.14 no.1
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    • pp.56-63
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    • 2010
  • Purpose : DIR image is relatively free from susceptibility artifacts therefore, DIR image can make it possible to reliably measure cortical thickness/volume. One drawback of the DIR acquisition is the long scan time to acquire the fully sampled 3D data set. To solve this problem, we applied a parallel imaging method (GRAPPA) and verify the reliability of using the volumetric study. Materials and methods : Six healthy volunteers (3 males and 3 females; age $25.33{\pm}2.25$ years) underwent MRI using the 3D DIR sequence at a 3.0T Siemens Tim Trio MRI scanner. GRAPPA simulation was performed from the fully sampled data set for reduction factor 2. Data reconstruction was performed using MATLAB R2009b. Freesurfer v.4.3.0 was used to evaluate the cortical thickness of the entire brain, and to extract white matter information from the DIR image, Analyze 9.0 was used. The global cortical thickness estimated from the reconstructed image was compared with reference image by using a T-test in SPSS. Results : Although reduced SNR and blurring are observed from the reconstructed image, in terms of segmentation the effect was not so significant. The volumetric result was validated that there were no significant differences in many cortical regions. Conclusion : This study was performed with DIR image for a volumetric MRI study. To solve the long scan time of 3D DIR imaging, we applied GRAPPA algorithm. According to the results, fast imaging can be done with reduction factor 2 with little loss of image quality at 3.0T.

Lactobacillus casei Secreting ${\alpha}$-MSH Induces the Therapeutic Effect on DSS-Induced Acute Colitis in Balb/c Mice

  • Yoon, Sun-Woo;Lee, Chul-Ho;Kim, Jeong-Yoon;Kim, Jie-Youn;Sung, Moon-Hee;Poo, Har-Young
    • Journal of Microbiology and Biotechnology
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    • v.18 no.12
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    • pp.1975-1983
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    • 2008
  • The neuropeptide ${\alpha}$-melanocyte-stimulating hormone (${\alpha}$-MSH) has anti-inflammatory property by down regulating the expressions of proinflammatory cytokines. Because ${\alpha}$-MSH elicits the anti-inflammatory effect in various inflammatory disease models, we examined the therapeutic effect of oral administration of recombinant Lactobacillus casei, which secretes ${\alpha}$-MSH (L. casei-${\alpha}$-MSH), on dextran sulfate sodium (DSS)-induced colitis in Balb/c mice. Thus, we constructed the ${\alpha}$-MSH-secreting Lactobacillus casei by the basic plasmid, pLUAT-ss, which was composed of a PldhUTLS promoter and ${\alpha}$-amylase signal sequence from Streptococcus bovis strain. Acute colitis was induced by oral administration of 5% DSS in drinking water for 7 days. To investigate the effect of L. casei-${\alpha}$-MSH on the colitis, L. casei or L. casei-${\alpha}$-MSH was orally administered for 7 days and their effects on body weight, mortality rate, cytokine production, and tissue myeloperoxidase (MPO) activity were observed. Administration of L. casei-${\alpha}$-MSH reduced the symptom of acute colitis as assessed by body weight loss (DSS alone: $14.45{\pm}0.2\;g$; L. casei-${\alpha}$-MSH: $18.2{\pm}0.12\;g$), colitis score (DSS alone: $3.6{\pm}0.4$; L. casei-${\alpha}$-MSH: $1.4{\pm}0.6$), MPO activity (DSS alone: $42.7{\pm}4.5\;U/g$; L. casei-${\alpha}$-MSH: $10.25{\pm}0.5\;U/g$), survival rate, and histological damage compared with the DSS alone mice. L. casei-${\alpha}$-MSH-administered entire colon showed reduced in vitro production of proinflammatory cytokines and $NF-{\kappa}B$ activation. The ${\alpha}$-MSH-secreting recombinant L. casei showed significant anti-inflammatory effects in the murine model of acute colitis and suggests a potential therapeutic role for this agent in clinical inflammatory bowel diseases.

Isolation of dhlA Gene Responsible for Degradation of 1, 2-dichloroethane from Metagenomic Library Derived from Daecheong Reservoir (대청호로부터 제작한 메타지놈 라이브러리에서 1, 2-dichloroethane의 분해에 관여하는 dhlA 유전자의 분리)

  • Kang, Cheol-Hee;Moon, Mi-Sook;Song, Ji-Sook;Lee, Sang-Mhan;Kim, Chi-Kyung
    • Korean Journal of Ecology and Environment
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    • v.38 no.2 s.112
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    • pp.137-145
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    • 2005
  • Traditional screening techniques have missed up to 99% of microbial resources existing in the nature. Strategies of direct cloning of environmental DNAs comprising tine genetic blueprints of entire microbial metagenomes provide vastly more genetic information than is contained in the culturable. Therefore, one way to screening the useful gene in a variety of environments is the construction of metagenomic DNA library. In this study, the water samples were collected from Daecheong Reservoir in the mid Korea, and analyzed by T-RFLP to examine the diversity of the microbial communities. The crude DNAs were extracted by SDS-based freezing-thawing method and then further purified using an $UltraClean^{TM}kit$ (MoBio, USA). The metagenomic libraries were constructed with the DNAs partially digested with EcoRI, BamHI, and SacII in Escherichia coli DH10B using the pBACe3.6 vector. About 14.0 Mb of metagenomic libraries were obtained with average inserts 13 ${\sim}$ 15 kb in size. The genes responsible for degradation of 1, 2-dichloroethane (1, 2-DCE) via hydrolytic dehalogenation were identified from the metagenomic libraries by colony hybridization. The 1, 2-dichloroethane dehalogenase gene (dhlA) was cloned and its nucleotide sequence was analyzed. The activity of the 1, 2-DCE dehalogenase was highly expressed to the substrate. These results indicated that the dhlA gene identified from the metagenomes derived from Deacheong Reservoir might be useful to develop a potent strain for degradation of 1, 2-DCE.

KMTNET SUPERNOVA PROGRAM VARIABLE OBJECTS I. NGC 2784 FIELD

  • HE, MATTHIAS YANG;MOON, DAE-SIK;NEILSON, HILDING;LEE, JAE-JOON;KIM, SANG CHUL;PAK, MINA;PARK, HONG SOO;KIM, DONG-JIN;LEE, YONGSEOK;KIM, SEUNG-LEE;LEE, CHUNG-UK
    • Journal of The Korean Astronomical Society
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    • v.49 no.5
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    • pp.209-223
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    • 2016
  • We present analyses of ~1250 variable sources identified in a 20 square degree field toward NGC 2784 by the KMTNet Supernova Program. We categorize the variable sources into three groups based on their B-band variability. The first group consists of 31 high variability sources with their B-band RMS variability greater than 0.3 magnitudes. The second group of medium variability contains 265 sources with RMS variability between 0.05 and 0.3 magnitudes. The remaining 951 sources belong to the third group of low variability with an RMS variability smaller than 0.05 magnitudes. Of the entire ~1250 sources, 4 clearly show periods of variability greater than 100 days, while the rest have periods shorter than ~51 days or no reliable periods. The majority of the sources show either rather irregular variability or short periods faster than 2 days. Most of the sources with reliable period determination between 2 and 51 days belong to the low-variability group, although a few belong to the medium-variability group. All the variable sources with periods longer than 35 days appear to be very red with B - V > 1.5 and V - I > 2.1 magnitudes. We classify candidates of 51 Cepheids, 17 semi-regular variables, 3 Mira types, 2 RV(B) Tauri stars, 26 eclipsing binary systems and 1 active galactic nucleus. The majority of long-term variables in our sample belong to either Mira or semi-regular types, indicating that long-term variability may be more prominent in post-main sequence phases of late-type stars. The depth of the eclipsing dips of the 26 candidates for eclipsing binaries is equivalent to ~0.61 as the average relative size of the two stars in the binary system. Our results illustrate the power of the KMTNet Supernova Program for future studies of variable objects.

A phylogenetic analysis of Korean Artemisia L. based on ITS sequences (ITS 염기서열에 의한 한국산 쑥속(Artemisia L.)의 계통분류학적 연구)

  • Lee, Jeong-Hoon;Park, Chung-Berm;Park, Chun-Geon;Moon, Sung-Gi
    • Korean Journal of Plant Resources
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    • v.23 no.4
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    • pp.293-302
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    • 2010
  • Taxa of Artemisia collected in Korea were constructed by molecular phylogenetic analysis based on the internal transcribed spacer(ITS) regions of nrDNA. The length of the ITS sequences aligned using the clustal X program was 636~643 bp, and the lengths of the ITS1 and ITS2 regions were 251~255 bp and 217~222 bp, respectively. The total number of variable sites was 95 for the entire sequence, and a parsimony- informative site represented an efficacious site in ITS1 rather than in ITS2. The maximum parsimony tree as calculated by the MEGA 4 program was clustered into five clades. The taxa(A. capillaris, A. japonica var. japonica, A. japonica var. hallaisanensis, A. japonica subsp. littoricora) degenerated ovary of clade 1 was supported as the subgenus Dracunculus by Ling's classification system. The results show that A. nakaii and A. fukudo were quite similar genetically(Boostrap 99%) and that the scientific name of Korean A. dubia should be reconsidered. A. sp. distributed in Ganghwa province was grouped with A. argyi(Boostrap 89%). These results suggest that the molecular techniques used in this study could be useful for the phylogenetic analysis of Korean Artemisia herbs having variations in their morphological characteristics.

Functional Genomics for Mass Analysis of Useful Genes in Panax ginseng C.A. Meyer (인삼의 유용유전자원 확보를 위한 기능 유전체연구)

  • Yang, Deok-Chun
    • Proceedings of the Ginseng society Conference
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    • 2004.05a
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    • pp.17-28
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    • 2004
  • As Korean ginseng is hybrid, an individual variation is very severe, and it takes long times in new breeding because it is required 4 years to pick the seed. But, transformation technique makes the high-functional breeding in short time. The focus of these ginseng studies is to find and secure the useful gene. And it is urgent to accumulate the fundamental data for the molecular breeding and secure the useful genes. Therefore, transformation and soil acclimatization technique are necessary to molecular breeding in use of the introduction of functional genes. In this study, it add to secure of new regulation gene and useful gene as to accumulate the fundamental data for the place where it will contribute to raise the national competitive power. To analyze the useful genes in large scale, we constructed CDNA libraries with various tissues, species, and treated tissue. EST analysis of ginseng perform in large scale and build the EST database of ginseng. We perform the full length sequencing about the selected lots of clones that include the entire open reading frame of the amino acid residues and construct cDNA chip with the parental EST clones. Establishment of the transformation and a soil acclimatization system throuth the re-introduction of the selected ginseng gene that related with the secondary metabolism and anti-stress into the ginseng.

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Changes of the Cellularities in the Bronchoalveolar Lavage Fluid of the Experimental Silicosis (흰쥐규폐증에서 기관지 폐포 세척액내 세포변화)

  • Lim, Young;Yun, Im-Goung;Oh, Min-Hwa
    • Tuberculosis and Respiratory Diseases
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    • v.38 no.2
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    • pp.143-154
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    • 1991
  • Pneumconiosis is a sort of pulmonary fibrosis consequent to the inhalation of the respirable dusts. Thus, the pathogenesis of silicosis have concentrated largely on the early response of alveolar macrophage and the later fibroblastic stimulation. But the role of the other cells and continuing cell injury in the pathogenesis has not been fully studied. And the chemical factors such as prostaglandin, fibroblast stimulating factor and inhibiting factor and chemotaxin are also participated in the mechanism of pulmonary fibrosis in silicosis. In order to clarify the role of alveolar cells and prostaglandin, we investigated the changes of the cellularities in bronchoalveolar lavage fluid and tissue pathology in the experimental silicosis with the time sequence. The experimental animals were divided into 3 groups; control group received only intratracheal injection of 0.5 ml saline, silica group received the intratracheal instillation of 40 mg silica with the same amount saline, and aspirin group received 450 mg/kg of aspirin after silica instillation. The results were as follows: 1) The total cells of bronchoalveolar lavage fluid in the silica group markedly increased in comparison with the control group, but there was no significant difference between the silica and aspirin groups. 2) The percentages of alveolar macrophages to the total number of cells in the silica group tended to be lower than those in the control group and also lower than those in the aspirin group at the 1st week after silica instillation. 3) The percentages of neutrophils to the total number of cells in the silica group were significantly higher than those in the control group during the entire period and also higher than those in the aspirin group at the 3rd day after silica instillation. 4) In the silica group, the percentages of lymphocytes to the total number of cells were increased 143 progressively with the time course and those were significantly higher than those in the control group from the 3rd week after silica administration. There were marked differences of lymphocyte percentages between the silica and aspirin groups at the 1st week after silica instillation. 5) The inflammatory change was observed in the rat lung at the 1st day after silica instillation. Also the silicotic nodule appeared in the silica group at the 1st week but we could not find out that nodule in the aspirin group at that time. The fibrotic changes in the rat lung tended to be increased progressively with the time course, therefore, the diffuse fibrotic pattern appeared in the whole field at the 20th week after silica instillation. 6) By the electron microscopy, there were gradual increases of phagosomes and vacuoles in the alveolar macrophage in the silica group as compared with the control group. These results suggest that the neutrophils and the lymphocytes have also participated in the pulmonary fibrosis even though the alveolar macrophage has a major role, and prostaglandin mediate the inflammation and pulmanary fibrosis in the experimental silicosis.

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A Microscope Study of Fluoride Effects on the Rat Incisor Enamel Formation (흰쥐 절치의 법랑질 형성에 미치는 불소의 효과에 관한 현미경적 연구)

  • Lim, Do-Seon;Chang, Byung-Soo;Jeong, Je-O;Jeong, Soon-Jeong;Jeong, Moon-Jin
    • Applied Microscopy
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    • v.34 no.2
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    • pp.145-157
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    • 2004
  • The aim of the present study was to examine in detail, both at light and electron microscopical levels, the morphological variations in ameloblast of the fetal rat incisor enamel organ. Rats were started on distilled water at the beginning of pregnancy. The pups were sacrificed 11 days after delivery and animals were perfused intravascularly with glutaraldehyde and the incisors were removed. To examine on the ultrastructure of the ameloblast, the study employed primary light microscopy but electron microscopy was used to clarify some of the light microscopic finding. Longitudinal sections through the incisors of the rat show a continuous layer of ameloblasts on the labial surface of the tooth. This layer contains the entire sequence of developmental stages in enamel production. The ameloblast layer was divided into three main zones: 1) Presecretory zone, region of ameloblasts facing pulp. 2) Secretory zone, region of inner and outer enamel secretion. 3) Maturation zone, region of reduced ameloblasts. In particularly, the present study has shown that two distinctively different types of ameloblasts appear in the enamel organ during enamel maturation in the rat incisor. These two types have been designated ruffle-ended ameloblasts (rAB) and smooth-ended ameloblasts (sAB). The fluoride produces marked alteration in the fine structure of ameloblast from teeth of young rats, such as large confluent distensions of the endoplasmic reticulum and swelling of isolated mitochondria. This experimental data suggested that exposure prolonged of animal to high level of fluoride appears to induce a few dramatic changes in the normal appositional growth and initial mineralization of enamel created during amelogenesis.