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ISOLATION AND IDENTIFICATION OF ANAEROBIC RUMEN BACTERIUM, ACTINOMYCES SP. 40 AND ENZYMATIC PROPERTIES OF β-1, 4-ENDOGLUCANASE

  • Min, H.K.;Choi, Y.J.;Ha, J.K.;Cho, K.K.;Kwon, Y.M.;Chang, Y.H.;Lee, S.S.
    • Asian-Australasian Journal of Animal Sciences
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    • v.7 no.3
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    • pp.373-382
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    • 1994
  • A bacterial strain No. 40, which produced extracellular endoglucanase, was isolated from the rumen of Korean native goals and identified to be a genus of Actinomyces sp. The optimum conditions for endoglucanase production in PY-CMC medium were initial pH of 7.0 and 4 days of cultivation at $39^{\circ}C$. When localization of endoglucanase activity of Actinomyces sp. was determined, 68% of the enzyme activity was found in the extracellular fraction, 11% of the activity was detected in the periplasmic space and the remaining activity was in the intracellular and cell-bound fractions. The maximal endoglucanase activity was observed at pH 5.0 and it was most s table at pH 5.0. The optimum temperature of this enzyme activity was $55^{\circ}C$, but enzyme activity was gradually lost at temperature above $60^{\circ}C$. The crude enzyme was activated by addition of 10 mM cysteine and 10 mM DTT. But it was inhibited by addition of 10 mM $Cu^{{+}{+}}$ and $Fe^{{+}{+}}$. This crude enzyme could digest carboxymethylcellulose (CMC), and degrade xylan, avicel, pNPG, and pNPC to a less extent.

Secretion of Ferritin Protein of Periserrula leucophyryna in Bacillus subtilis and Its Feed Efficiency (고초균에서 흰이빨참갯지렁이 페리틴 단백질의 분비 및 사료 효율성)

  • Choi, Jang Won
    • KSBB Journal
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    • v.31 no.2
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    • pp.105-112
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    • 2016
  • Ferritin is known to regulate iron metabolism and maintain iron in a variety of the eukaryotic organisms. The region encoding the mature ferritin (0.47 kb, H-type) of Periserrula leucophryna was amplified using the designed primers including restriction enzyme site and termination codon and subcloned in frame to the pRBAS secretion vector containing the signal sequence, RBS, and promoter of amylase gene (E. coli-Bacillus shuttle vector), resulting in recombinant pRBAS-PLF vector. Recombinant ferritin (18 kDa) was correctly processed and secreted from Bacillus subtilis LKS strain harboring the pRBAS-PLF vector and quantitatively analyzed by SDS-PAGE and western blot, respectively. Secretion of the ferritin was optimized by culture conditions (host, medium, temperature, nitrogen source) in 3 L batch culture and 5 L jar fermenter. Finally. the ferritin was largely produced using 50 L fermenter as the following conditions; at $30^{\circ}C$, 150 rpm, 1 vvm in Bacillus subtilis LKS using PY medium. The secreted ferritin was maximally measured (approximately 177.6 ug/ml) when the cell density reached to 14.4 at $OD_{600}$ (20 h incubation). The iron binding activity was confirmed by Perls' staining in 7.5% non-denaturing gel, indicating that the multimeric ferritin (composed of 24 subunits) was formed in the culture broth after secretion. Biologically, the culture broth and powder type containing ferritin were tested for possibility as feed additive in chicken broiler. As a result, the ferritin stimulated the growth of chick broil and improved feed efficiency and production index.

Development of Micro Tensile Test of CVD-SiC coating Layer for TRISO Nuclear Fuel Particles at elevated temperature

  • Lee, Hyun-Min;Park, Kwi-Il;Kim, Do-Kyung
    • Proceedings of the Materials Research Society of Korea Conference
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    • 2012.05a
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    • pp.95.1-95.1
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    • 2012
  • Very High Temperature gas cooler Reactor (VHTR) has been considered as one of the most promising nuclear reactor because of many advantages including high inherent safety to avoid environmental pollution, high thermal efficiency and the role of secondary energy source. The TRISO coated fuel particles used in VHTR are composed of 4 layers as OPyC, SiC, IPyC and buffer PyC. The significance of CVD-SiC coatings used in tri-isotropic(TRISO) nuclear coated fuel particles is to maintain the strength of the whole particle. Various methods have been proposed to evaluate the mechanical properties of CVD-SiC film at room temperature. However, few works have been attempted to characterize properties of CVD-SiC film at high temperature. In this study, micro tensile system was newly developed for mechanical characterization of SiC thin film at elevated temperature. Two kinds of CVD-SiC films were prepared for micro tensile test. SiC-A had [111]-preferred orientation, while SiC-B had [220]-preferred orientation. The free silicon was co-deposited in SiC-B coating layer. The fracture strength of two different CVD-SiC films was characterized up to $1000^{\circ}C$.The strength of SiC-B film decreased with temperature. This result can be explained by free silicon, observed in SiC-B along the columnar boundaries by TEM. The presence of free silicon causes strength degradation. Also, larger Weibull-modulus was measured. The new method can be used for thin film material at high temperature.

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The Secretion Optimization of Oligopeptide with His-Pro Repeats in Bacillus subtilis and Its Anti-Diabetic Effects (고초균에서 His-Pro 반복서열을 갖는 Oligopeptide의 분비 최적화 및 항당뇨 효과)

  • Jeong, Seon Hwa;Choi, Jang Won
    • KSBB Journal
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    • v.32 no.1
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    • pp.71-82
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    • 2017
  • To verify anti-diabetic effect of oligopeptide with His-Pro repeats (mHP peptide), the oligopeptide was first secreted and optimized using the secretion vector, pRBAS with alkaline protease gene promoter and the signal sequence in Bacillus subtilis and directly the anti-diabetic effect of the mHP peptide was investigated in insulinoma cell, RINm5F cell line. The oligopeptide gene was obtained by annealing oligonucleotides with repeated His-Pro sequence and finally was constructed as 18 dipeptides (108 bp and 4.0 kDa) coding gene, named oligopeptide with His-Pro repeats (mHP peptide) to make cyclo(His-Pro) known to be anti-diabetic effects. The region encoding the oligopeptide gene was subcloned into the pRBAS secretion vector (E.coli-Bacillus shuttle vector) after PCR amplification using the designed primers including initiation and termination codons and His tag, named pRBAS-mHP (6.56 kb). To optimize secretion of the oligopeptide, various culture conditions were investigated in Bacillus subtilis LKS. As a result, the secreted oligopeptide was maximally measured (approximately $59.6{\mu}g/mL$) in 3 L batch culture and the highest secretion was achieved at $30^{\circ}C$, PY medium, and carbon sources (particularly barley and glycerol). In the RINm5F cells treated with 2 mM STZ, the oligopeptide treatment (0.1 mg/mL) restored the cell viability (10%) and reduced the nitric oxide (NO) generation (35%) and DNA fragmentation (90%). And also, insulin secretion level was increased to 17% higher than in STZ-treated RINm5F cells. These results suggest that the oligopeptide with His-Pro repeats could be a candidate material for anti-diabetic agent against STZ-induced diabetes.

Non-restraint Master Interface of Minimally Invasive Surgical Robot Using Hand Motion Capture (손동작 영상획득을 이용한 최소침습수술로봇 무구속 마스터 인터페이스)

  • Jang, Ik-Gyu
    • Journal of Biomedical Engineering Research
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    • v.37 no.3
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    • pp.105-111
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    • 2016
  • Introduction: Surgical robot is the alternative instrument that substitutes the difficult and precise surgical operation; should have intuitiveness operationally to transfer natural motions. There are limitations of hand motion derived from contacting mechanical handle in the surgical robot master interface such as mechanical singularity, isotropy, coupling problems. In this paper, we will confirm and verify the feasibility of intuitive Non-restraint master interface which tracking the hand motion using infra-red camera and only 3 reflective markers without the hardware handle for the surgical robot master interface. Materials & methods: We configured S/W and H/W system; arranged 6 infra-red cameras and attached 3 reflective markers on hands for measuring 3 dimensional coordinate then we find the 7 motions of grasp, yaw, pitch, roll, px, py, pz. And we connected Virtual-Master to the slave surgical robot(Laparobot) and observed the feasibility. To verify the result of motion, we compare the result of Non-restraint master and that of clinometer (and protractor) through measuring 0~180 degree, 10degree interval, 1000 samples and recorded standard deviation stands for error rate of the value. Results: We confirmed that the average angle values of Non-restraint master interface is accurately corresponds to the result of clinometer (and protractor) and have low error rates during motion. Investigation & Conclusion: In this paper, we confirmed the feasibility and accuracy of 3D Non-restraint master interface that can offer the intuitive motion of non-contact hardware handle. As a result, we can expect the high intuitiveness, dexterousness of surgical robot.

Identification of the Antibiotic-Producing Chstridium sp. KH-431 and Purification of the Antibiotics (항생물질을 생산하는 Clostridium sp. KH-431의 동정 및 항생물질의 정제)

  • 홍수형;김미정;박용복;이재근;하지홍
    • Microbiology and Biotechnology Letters
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    • v.21 no.1
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    • pp.41-46
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    • 1993
  • A strain showing antibiotic activities against various bacteria and fungi was selected from approximately 2,000 microorganisms obtained from soil samples. This strain, designated as KH-431, was identified as a Clostridium sp. by its morphological, physiological and biochemical characteristics. The highest production of the antibiotics was achieved in a fermentation medium containing sorbitol, yeast extract, d-biotin and $CaCl_2$ The antibiotics were isolated from the culture broth by solvent extraction using ethyl acetate, silica gel column chromatography and recrystallization. Two kinds of antibiotics, KG-431A and KG-431B were obtained after the purification procedure, and only KG-431B was successful to recrystallize.

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Effect of Epigallocatechin Gallate on Inhibition of Cell Proliferation in MDA-MB-231 Human Breast Cancer Cells (Epigallocatechin Gallate가 인체 유방암 세포인 MDA-MB-231의 세포증식억제에 미치는 영향)

  • Hong, Eun-Jung;Kim, Woo-Kyung
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.36 no.8
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    • pp.983-988
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    • 2007
  • Epigallocatechin gallate (EGCG), a principal antioxidant derived from green tea, is one of the most extensively investigated chemopreventive phytochemicals. However, the effect of EGCG on proliferation in MDA-MB-231 breast cancer cell is not well known. We investigated the effect of EGCG on protein and mRNA expression related to cell proliferation in MDA-MB-231 human breast cancer cell lines. We cultured MDA-MB-231 cells in the presence of 0, 5, 10 and 20 ${\mu}m$ of EGCG. EGCG significantly inhibited the cancer cell proliferation (p<0.05). In MDA-MB-231 huamn breast cancer cell, EGCG lowered $ErbB_2$ and $ErbB_3$ protein as well as mRNA expression. In addition, protein and mRNA expression of phosphorylated Akt and total Akt were significantly decreased (p<0.05). We suggest that EGCG inhibits cell proliferation through $ErbB_2$, $ErbB_3$ and Akt cell signaling.

Quantitative Evaluation of Radiation Dose Rates for Depleted Uranium in PRIDE Facility

  • Cho, Il Je;Sim, Jee Hyung;Kim, Yong Soo
    • Journal of Radiation Protection and Research
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    • v.41 no.4
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    • pp.378-383
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    • 2016
  • Background: Radiation dose rates in PRIDE facility is evaluated quantitatively for assessing radiation safety of workers because of large amounts of depleted uranium being handled in PRIDE facility. Even if direct radiation from depleted uranium is very low and will not expose a worker to significant amounts of external radiation. Materials and Methods: ORIGEN-ARP code was used for calculating the neutron and gamma source term being generated from depleted uranium (DU), and the MCNP5 code was used for calculating the neutron and gamma fluxes and dose rates. Results and Discussion: The neutron and gamma fluxes and dose rates due to DU on spherical surface of 30 cm radius were calculated with the variation of DU mass and density. In this calculation, an imaginary case in which DU density is zero was added to check the self-shielding effect of DU. In this case, the DU sphere was modeled as a point. In case of DU mixed with molten salt of 50-250 g, the neutron and gamma fluxes were calculated respectively. It was found that the molten salt contents in DU had little effect on the neutron and the gamma fluxes. The neutron and the gamma fluxes, under the respective conditions of 1 and 5 kg mass of DU, and 5 and $19.1g{\cdot}cm^{-3}$ density of DU, were calculated with the molten salt (LiCl+KCl) of 50 g fixed, and compared with the source term. As the results, similar tendency was found in neutron and gamma fluxes with the variation of DU mass and density when compared with source spectra, except their magnitudes. Conclusion: In the case of the DU mass over 5 kg, the dose rate was shown to be higher than the environmental dose rate. From these results, it is concluded that if a worker would do an experiment with DU having over 5 kg of mass, the worker should be careful in order not to be exposed to the radiation.

Virtual Screening for Potential Inhibitors of NS3 Protein of Zika Virus

  • Sahoo, Maheswata;Jena, Lingaraja;Daf, Sangeeta;Kumar, Satish
    • Genomics & Informatics
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    • v.14 no.3
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    • pp.104-111
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    • 2016
  • Zika virus (ZIKV) is a mosquito borne pathogen, belongs to Flaviviridae family having a positive-sense single-stranded RNA genome, currently known for causing large epidemics in Brazil. Its infection can cause microcephaly, a serious birth defect during pregnancy. The recent outbreak of ZIKV in February 2016 in Brazil realized it as a major health risk, demands an enhanced surveillance and a need to develop novel drugs against ZIKV. Amodiaquine, prochlorperazine, quinacrine, and berberine are few promising drugs approved by Food and Drug Administration against dengue virus which also belong to Flaviviridae family. In this study, we performed molecular docking analysis of these drugs against nonstructural 3 (NS3) protein of ZIKV. The protease activity of NS3 is necessary for viral replication and its prohibition could be considered as a strategy for treatment of ZIKV infection. Amongst these four drugs, berberine has shown highest binding affinity of -5.8 kcal/mol and it is binding around the active site region of the receptor. Based on the properties of berberine, more similar compounds were retrieved from ZINC database and a structure-based virtual screening was carried out by AutoDock Vina in PyRx 0.8. Best 10 novel drug-like compounds were identified and amongst them ZINC53047591 (2-(benzylsulfanyl)-3-cyclohexyl-3H-spiro[benzo[h]quinazoline-5,1'-cyclopentan]-4(6H)-one) was found to interact with NS3 protein with binding energy of -7.1 kcal/mol and formed H-bonds with Ser135 and Asn152 amino acid residues. Observations made in this study may extend an assuring platform for developing anti-viral competitive inhibitors against ZIKV infection.

Synthesis and Characterization of Iridium-Containing Green Phosphorescent Polymers for PLEDs

  • Xu, Fei;Kim, Hee Un;Mi, Dongbo;Lim, Jong Min;Hwang, Ju Hyun;Cho, Nam Sung;Lee, Jeong-Ik;Hwang, Do-Hoon
    • Bulletin of the Korean Chemical Society
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    • v.34 no.2
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    • pp.399-405
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    • 2013
  • Two series of new green phosphorescent polymers bearing a bis(2-phenyl-pyridine)iridium(III)(dibenzoylmethane) [$(ppy)_2Irdbm$] complex were designed and synthesized. Poly-carbazole (PCbz) derivative or polyfluorene with pendant carbazole groups (PFCbz) were employed as host polymers for the iridium complex. The iridium complex monomer was copolymerized with the host monomers using varying monomer ratios via a Yamamoto coupling reaction. Efficient energy transfer from host to dopant unit was observed by increasing the ratio of the iridium guest in the copolymers. Electroluminescent devices with the configuration ITO/PEDOT:PSS/polymer/BmPyPB/LiF/Al were fabricated and characterized. The phosphorescent polymers composed of the iridium complex guest and polyfluorene with carbazole pendants as a host performed better than the polymers composed of the same guest and the main chain polycarbazole host. A maximum external quantum efficiency of 0.73%, a luminous efficiency of 1.21 cd/A, and a maximum luminance of 372 $cd/m^2$ were obtained from a device fabricated using one of the synthesized copolymers.