• 제목/요약/키워드: Helical

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Directed Mutagenesis of the Bacillus thuringiensis Cry11A Toxin Reveals a Crucial Role in Larvicidal Activity of Arginine-136 in Helix 4

  • Angsuthanasombat, Chanan;Keeratichamreon, Siriporn;Leetacheewa, Somphob;Katzenmeier, Gerd;Panyim, Sakol
    • BMB Reports
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    • v.34 no.5
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    • pp.402-407
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    • 2001
  • Based on the currently proposed toxicity model for the different Bacillus thuringiensis Cry $\delta$-endotoxins, their pore-forming activity involves the insertion of the ${\alpha}4-{\alpha}5$ helical hairpin into the membrane of the target midgut epithelial cell. In this study, a number of polar or charged residues in helix 4 within domain I of the 65-kDa dipteranactive Cry11A toxin, Lys-123, Tyr-125, Asn-128, Ser-130, Gln-135, Arg-136, Gln-139 and Glu-141, were initially substituted with alanine by using PCR-based directed mutagenesis. All mutant toxins were expressed as cytoplasmic inclusions in Escherichia coli upon induction with IPTG. Similar to the wild-type protoxin inclusion, the solubility of each mutant inclusion in the carbonate buffer, pH 9.0, was relatively low When E. coli cells, expressing each of the mutant proteins, were tested for toxicity against Aedes aegypti mosquito-larvae, toxicity was completely abolished for the alanine substitution of arginine at position 136. However, mutations at the other positions still retained a high level of larvicidal activity Interestingly, further analysis of this critical arginine residue by specific mutagenesis showed that conversions of arginine-136 to aspartate, glutamine, or even to the most conserved residue lysine, also abolished the wild-type activity The results of this study revealed an important determinant in toxin function for the positively charged side chain of arginine-136 in helix 4 of the Cry11A toxin.

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Numerical analysis of turbulent flows in the helically coiled pipes of heat transfer (열교환기의 나선형 관내 난류유동 수치해석)

  • Kwag, Seung-Hyun
    • Journal of Advanced Marine Engineering and Technology
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    • v.37 no.8
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    • pp.905-910
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    • 2013
  • The flow analysis has been made by applying the turbulent models in the helically coiled tubes of heat transfer. The k-${\varepsilon}$ and Spalart-Allmaras turbulent models are used in which the structured grid is applied for the simulation. The velocity vector, the pressure contour, the change of residuals along the iteration number and the friction factors are simulated by solving the Navier-Stokes equations to make clear the Reynolds number effect. The helical tube increases the centrifugal forces by which the wall shear stress become larger on the outer side of the tube. The centrifugal force makes the heat transfer rate locally larger due to the increase of the flow energy, which finds out the close relationship between the pressure drop and friction factor in the internal flow. The present numerical results are compared with others, for example, in the value of friction factor for validation.

A HISTOLOGIC AND AUTORADIOGRAPHIC STUDY ON THE EARLY CHANGES IN THE MOVEMENTS OF RAT INCISORS (백서의 전치 이동시 초기 변화에 관한 조직학적 및 자기방사법적 연구)

  • Yoo, Ki-Whan;Kim, Sang-cheol;Kook, Yoon-A
    • The korean journal of orthodontics
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    • v.23 no.2 s.41
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    • pp.199-216
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    • 1993
  • The purpose of this study was to quantify the biologic effects of the tensile forces from helical springs across the maxillary incisors on the periodontal tissues of rats. 39 Sprague-Dawlely rats were divided into a control group(3 rats) and three experimental groups(36 rats)-group 1, pressured with a light force(50-75g), group 2, with a heavy force(250-300g) and group 3, with a hen force(250-300g) plus laser irradiation. Autoradiographic and histopathologic observations were performed in 12, 24, 48 and 96 hours after force application. The result were as follow : 1. Hyalinized zone of periodontal ligament began to appear at pressure side in 12 hours in group 2 and group 3 ; all decreased in 96 hours except in group 2. 2. Alveolar bone resorption began to appear in 12 hours in group 2 and group 3 ; Group 2 showed more resorption than group 1 and no difference to group 3. 3. Tearing of periodontal ligament and vascular dilatation began to appear at tension side in 12 hours in all groups ; Group 2 showed more changes than group 1 and no difference to group 3 ; Decrease began to appear in 96 hours. 4. New bone formation began to appear at tension side in 12 hours and increased more and more ; No differences were shown of groups 5. New capillary proliferation began to appear at pressure side in 12 hours ; The changes were the greatest in group 3, group 2, group 1, in that order. 6. Positive reaction of cells to $[^3H]-thymidine$ was the greatest in 24 hours of all groups and decreased with times i Group 2 showed more reaction than group 1 and no difference to group 3.

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Identification of Antimicrobial Peptide Hexamers against Oral Pathogens through Rapid Screening of a Synthetic Combinatorial Peptide Library

  • Song, Je-Seon;Cho, Kyung Joo;Kim, Joungmok;Kim, Jeong Hee
    • International Journal of Oral Biology
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    • v.39 no.4
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    • pp.169-176
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    • 2014
  • A positional scanning synthetic peptide combinatorial library (PS-SCL) was screened in order to identify antimicrobial peptides against the cariogenic oral bacteria, Streptococcus mutans. Activity against Streptococcus gordonii and Aggregatibacter actinomycetemcomitans was also examined. The library was comprised of six sub-libraries with the format $O_{(1-6)}XXXXX-NH_2$, where O represents one of 19 amino acids (excluding cysteine) and X represents equimolar mixture of these. Each sub-library was tested for antimicrobial activity against S. mutans and evaluated for antimicrobial activity against S. gordonii and A. actinomycetemcomitans. The effect of peptides was observed using transmission electron microscopy (TEM). Two semi-mixture peptides, RXXXXN-$NH_2$ (pep-1) and WXXXXN-$NH_2$ (pep-2), and one positioned peptide, RRRWRN-$NH_2$ (pep-3), were identified. Pep-1 and pep-2 showed significant antimicrobial activity against Gram positive bacteria (S. mutans and S. gordonii), but not against Gram negative bacteria (A. actinomycetemcomitans). However, pep-3 showed very low antimicrobial activity against all three bacteria. Pep-3 did not form an amphiphilic ${\alpha}$-helix, which is a required structure for most antimicrobial peptides. Pep-1 and pep-2 were able to disrupt the membrane of S. mutans. Small libraries of biochemically-constrained peptides can be used to generate antimicrobial peptides against S. mutans and other oral microbes. Peptides derived from such libraries may be candidate antimicrobial agents for the treatment of oral microorganisms.

Replication origins oriGNAI3 and oriB of the mammalian AMPD2 locus nested in a region of straight DNA flanked by intrinsically bent DNA sites

  • Balani, Valerio Americo;De Lima Neto, Quirino Alves;Takeda, Karen Izumi;Gimenes, Fabricia;Fiorini, Adriana;Debatisse, Michelle;Fernandez, Maria Aparecida
    • BMB Reports
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    • v.43 no.11
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    • pp.744-749
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    • 2010
  • The aim of this work was to determine whether intrinsically bent DNA sites are present at, or close to, the mammalian replication origins oriGNAI3 and oriB in the Chinese hamster AMPD2 locus. Using an electrophoretic mobility shift assay and in silico analysis, we located four intrinsically bent DNA sites (b1 to b4) in a fragment that contains the oriGNAI3 and one site (b5) proximal to oriB. The helical parameters show that each bent DNA site is curved in a left-handed superhelical writhe. A 2D projection of 3D fragment trajectories revealed that oriGNAI3 is located in a relatively straight segment flanked by bent sites b1 and b2, which map in previously identified Scaffold/Matrix Attachment Region. Sites b3 and b4 are located approximately 2 kb downstream and force the fragment into a strong closed loop structure. The b5 site is also located in an S/MAR that is found just downstream of oriB.

Design and Construction of a Loom for Obtaining Ultra-Light Metal Structure (초경량 금속 구조재 직조장치의 설계 및 제작)

  • Kim, Pan-Su;Kang, Ki-Ju
    • Transactions of the Korean Society of Mechanical Engineers A
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    • v.34 no.9
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    • pp.1235-1240
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    • 2010
  • Wire-woven Bulk Kagome (WBK) is fabricated by assembling helically formed wires in six directions. To date, WBK samples have been assembled manually. For industrial application, the assembly process must be automated. Furthermore, if WBK is to be fabricated using flexible wires that cannot maintain their helical shape during fabrication, a specialized automatic machine, i.e., a loom needs to be developed. In this work, we designed and constructed a loom for fabricating WBKs using flexible wires. This loom is operated by one rotation of the upper plate, two translations of the insertion device, and insertion of wires. So-called "comb devices" are placed between multiple layers of Kagome nets to prevent the wires that are already in place from getting entangled with those that are being inserted. This loom can be also used to fabricate semi-WBKs composed of helically formed wires and rigid straight wires.

Discharge Characteristics of Large-Area High-Power RF Ion Source for Neutral Beam Injector on Fusion Devices

  • Chang, Doo-Hee;Park, Min;Jeong, Seung Ho;Kim, Tae-Seong;Lee, Kwang Won;In, Sang Ryul
    • Proceedings of the Korean Vacuum Society Conference
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    • 2014.02a
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    • pp.241.1-241.1
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    • 2014
  • The large-area high-power radio-frequency (RF) driven ion sources based on the negative hydrogen (deuterium) ion beam extraction are the major components of neutral beam injection (NBI) systems in future large-scale fusion devices such as an ITER and DEMO. Positive hydrogen (deuterium) RF ion sources were the major components of the second NBI system on ASDEX-U tokamak. A test large-area high-power RF ion source (LAHP-RaFIS) has been developed for steady-state operation at the Korea Atomic Energy Research Institute (KAERI) to extract the positive ions, which can be used for the NBI heating and current drive systems in the present fusion devices, and to extract the negative ions for negative ion-based plasma heating and for future fusion devices such as a Fusion Neutron Source and Korea-DEMO. The test RF ion source consists of a driver region, including a helical antenna and a discharge chamber, and an expansion region. RF power can be transferred at up to 10 kW with a fixed frequency of 2 MHz through an optimized RF matching system. An actively water-cooled Faraday shield is located inside the driver region of the ion source for the stable and steady-state operations of RF discharge. The characteristics and uniformities of the plasma parameter in the RF ion source were measured at the lowest area of the expansion bucket using two RF-compensated electrostatic probes along the direction of the short- and long-dimensions of the expansion region. The plasma parameters in the expansion region were characterized by the variation of loaded RF power (voltage) and filling gas pressure.

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Thermotropic Liquid Crystalline Behavior of Poly(1-cholesteryloxycarbonyloxy]ethylene] and Poly[1-(cholesteryloxycarbonylheptanoyloxy)ethylene] (폴리[1-(콜레스테릴옥시카보닐옥시)에틸렌]과 폴리[1-(콜레스테릴옥시카보닐헵타노일옥시)에틸렌]의 열방성 액정 거동)

  • Jeong, Seung-Yong;Ma, Yung-Dae
    • Polymer(Korea)
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    • v.30 no.1
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    • pp.35-44
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    • 2006
  • Poly[1-(cholesteryloxycarbonyloxy)ethylene](PCOE) and poly[1-(cholesteryloxycarbonylheptanoyloxy)ethylene] (PCOSE) were prepared by reacting poly(vinyl alcohol) with cholesteryl chloroformate or 8-cholesteryloxycarbonylheptanoly] chloride (CH8C), and their thermal and optical properties were investigated. CH8C formed a monotropic cholesteric phase whereas PCOE and PCOSE exihibited enantiotropic cholesteric phases. Like in the case of CH8C, the optical pitch $(\lambda_m)$ of PCOSE decreased with increasing temperature. PCOE, contrast with PCOSE, did not display reflection colors, suggesting that the helical twisting power or the cholesteryl group highly depends on the length or the spacer joining the cholesteryl group to the main chain. The mesophase properties of PCOE and PCOSE were entirely different from those of poly $(cholesteryl-\omega-acryloyloxyalkanoates)$. The results indicate that the mode of chemical linkage of the side chain group with the main chain plays an important role in the formation, stabilization, and temperature dependence of $\lambda_m$ of the cholesteric mesophase.

Study on the Process to Decrease the molecular Weight of $\beta$-[1,6]-branched $\beta$-[1,3]-D-Glucans (분지 베타 글루칸의 저분자화 기술 연구)

  • 신현재;이동철
    • KSBB Journal
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    • v.18 no.5
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    • pp.352-355
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    • 2003
  • ${\beta}$-(1,6)-Branched ${\beta}$-(1,3)-D-glucans are known to enhance the immune system in human body, and in most cases have higher molecular weights over 1 MDa. In order to enhance the efficacy of glucans by decreasing their molecular weights, sonication, acid treatment, and enzymatic hydrolysis were tested and compared in this work. Treatment of sonication was effective to decrease the molecular weight to the extent of several dozens of kilo-daltons, but have a risk to disorder the triple helical structure of the glucans. Acid treatment was also an effective method to degrade polysaccharides, but ${\beta}$-(1,6)-branched of the glucan molecules was found to be also hydrolyzed. Treatment of ${\beta}$-(1,3)-glucanase was an effective method to decrease the molecular weight in mild conditions, but could not hydrolyse the highly ${\beta}$-(1,6)-branched ${\beta}$-(1,3)-glucans efficiently.

Axisymmetric Swirling Flow Simulation of the Draft Tube Vortex in Francis Turbines at Partial Discharge

  • Susan-Resiga, Romeo;Muntean, Sebastian;Stein, Peter;Avellan, Francois
    • International Journal of Fluid Machinery and Systems
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    • v.2 no.4
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    • pp.295-302
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    • 2009
  • The flow in the draft tube cone of Francis turbines operated at partial discharge is a complex hydrodynamic phenomenon where an incoming steady axisymmetric swirling flow evolves into a three-dimensional unsteady flow field with precessing helical vortex (also called vortex rope) and associated pressure fluctuations. The paper addresses the following fundamental question: is it possible to compute the circumferentially averaged flow field induced by the precessing vortex rope by using an axisymmetric turbulent swirling flow model? In other words, instead of averaging the measured or computed 3D velocity and pressure fields we would like to solve directly the circumferentially averaged governing equations. As a result, one could use a 2D axi-symmetric model instead of the full 3D flow simulation, with huge savings in both computing time and resources. In order to answer this question we first compute the axisymmetric turbulent swirling flow using available solvers by introducing a stagnant region model (SRM), essentially enforcing a unidirectional circumferentially averaged meridian flow as suggested by the experimental data. Numerical results obtained with both models are compared against measured axial and circumferential velocity profiles, as well as for the vortex rope location. Although the circumferentially averaged flow field cannot capture the unsteadiness of the 3D flow, it can be reliably used for further stability analysis, as well as for assessing and optimizing various techniques to stabilize the swirling flow. In particular, the methodology presented and validated in this paper is particularly useful in optimizing the blade design in order to reduce the stagnant region extent, thus mitigating the vortex rope and expending the operating range for Francis turbines.