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Secretory Expression and Purification of the Recombinant Duck Interleukin-2 in Pichia pastoris

  • Du, Cuihong;Han, Long;Xiao, Anfeng;Cao, Minjie
    • Journal of Microbiology and Biotechnology
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    • v.21 no.12
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    • pp.1264-1269
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    • 2011
  • Interleukin-2 (IL-2) is a vital cytokine secreted by activated T lymphocytes, and plays an important role in the regulation of cellular functions and immunity of animals. In this study, the recombinant duck IL-2 (rduIL-2) was secretory expressed in Pichia pastoris (P. pastoris). The recombinant P. pastoris strain was cultured in shake flasks and then scaled up in a 5.0-l bioreactor. The result showed that the maximal fresh-cell-weight of 594.1 g/l and the maximal $OD_{600}$ of 408 were achieved in the bioreactor. The rduIL-2 was purified by two steps of purification procedures, and approximately 311 mg of rduIL-2/L fermentation supernatant was obtained. SDS-PAGE showed that the purified rduIL-2 constituted a homogeneous band of ~16 kDa or ~14 kDa corresponding to the glycosylated or non-glycosylated duIL-2 protein in size, respectively. The bioactivity of rduIL-2 was determined by lymphocyte proliferation assay. The result indicated that the rduIL-2 greatly promoted the proliferation of ConA-stimulated lymphocytes in vitro. The P. pastoris expression system described here could provide promising, inexpensive, and large-scale production of the rduIL-2, which lays the foundation for development of novel immunoadjuvants to enhance both the immunity of ducks against various infectious pathogens and vaccine efficacy.

Application of Friction Stir Process to Improve Surface Reliability of Light Weight Magnesium Alloy (경량 마그네슘 합금의 표면 신뢰성 향상을 위한 마찰교반공정의 적용)

  • Gil, Ung-Chan;Kim, Jae-Yeon;Hyun, Chang-Young
    • Journal of Applied Reliability
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    • v.16 no.2
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    • pp.155-161
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    • 2016
  • Purpose: Purpose of this study is to analyze the effect of particle size as well as number of pass on surface microstructure and hardness of SiC(p)/AZ31 surface composite fabricated by friction stir process (FSP). Method: SiC(p)/AZ31 surface composite containing different size of SiC particle (i. e., $2{\mu}m$ and $8{\mu}m$) was fabricated by multi-pass FSP. Microstructure was observed by scanning electron microscope and surface hardness was determined by Vickers hardness tester. Results: For all the FSPed specimens with and without hardening particles, grain size was refined due to dynamic recrystallization behavior. Surface hardness was observed to increase with decreasing particle size in the composite layer. Increasing number of FSP pass was effective for homogeneous distribution of the hardening particles and for resulting increase in surface hardness. Conclusion: FSP was effective to modify surface microstructure for improving surface hardness of SiC/AZ31 composite.

Time-dependent and inelastic behaviors of fiber- and particle hybrid composites

  • Kim, Jeong-Sik;Muliana, Anastasia
    • Structural Engineering and Mechanics
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    • v.34 no.4
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    • pp.525-539
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    • 2010
  • Polymer matrix composites are widely used in many engineering applications as they can be customized to meet a desired performance while not only maintaining low cost but also reducing weight. Polymers can experience viscoelastic-viscoplastic response when subjected to external loadings. Various reinforcements and fillers are added to polymers which bring out more complexity in analyzing the timedependent response. This study formulates an integrated micromechanical model and finite element (FE) analysis for predicting effective viscoelastic-viscoplastic response of polymer based hybrid composites. The studied hybrid system consists of unidirectional short-fiber reinforcements and a matrix system which is composed of solid spherical particle fillers dispersed in a homogeneous polymer constituent. The goal is to predict effective performance of hybrid systems having different compositions and properties of the fiber, particle, and matrix constituents. A combined Schapery's viscoelastic integral model and Valanis's endochronic viscoplastic model is used for the polymer constituent. The particle and fiber constituents are assumed linear elastic. A previously developed micromechanical model of particle reinforced composite is first used to obtain effective mechanical properties of the matrix systems. The effective properties of the matrix are then integrated to a unit-cell model of short-fiber reinforced composites, which is generated using the FE. The effective properties of the matrix are implemented using a user material subroutine in the FE framework. Limited experimental data and analytical solutions available in the literatures are used for comparisons.

Purification and Characterization of ${\alpha}$-L-Arabinosidase from Trichoderma sp. SY

  • Jung, Bo-Ra;Kim, Bong-Gyu;Lee, Yoon-Jung;Ahn, Joong-Hoon
    • Journal of Applied Biological Chemistry
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    • v.48 no.1
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    • pp.7-10
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    • 2005
  • Trichoderma sp. SY most effectively produces an extracellular ${\gamma}$-L-arabinofuranosidase (AF) using arabinose as a carbon source. AF grown on cellulose as a carbon source was purified 28-fold with 4.4% yield by DEAE exchange and HQ/20 cation exchange chromatographies The purified enzyme was found to be homogeneous on SDS-PAGE with molecular weight of 89 kDa. It exhibited a high level of activity with p-nitrophenyl ${\alpha}$-L-arabinofuranoside, showing $K_m$ and $V_{max}$ values of $0.15\;{\mu}M$ and $239.85U{\cdot}mg^{-1}$, respectively and did not require any metal ion for activity. It also released p-nitrophenol from p-nitrophenol conjugated ${\beta}$-D-xylopyranoside, and ${\beta}$-D-galactopyranoside not from ${\beta}$-D-glucopyranoside.

Novel Cationic Microbial Polyglucosamine Biopolymer from New Enterobacter sp. BL-2 and Its Bioflocculation Efficacy

  • SON MI-KYUNG;SHIN HYUN-DONG;HUH TAE-LIN;JANG JIN-HO;LEE YONG-HYUN
    • Journal of Microbiology and Biotechnology
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    • v.15 no.3
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    • pp.626-632
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    • 2005
  • A new bacterium BL-2 excreting a novel cationic polyglucosamine biopolymer was isolated from the spoiled leaves of Chinese cabbage and identified as Enterobacter sp. BL-2. The isolated Enterobacter sp. BL-2 was cultivated in pH-stat fed-batch culture using acetic acid as the feeding stock at pH 8.0, resulting in 17.11 g/l of cells and 1.53 g/l of an extracellular biopolymer after 72 h. The excreted biopolymer was purified by a three-step procedure, involving ethanol precipitation and deproteinizations, to a nearly homogeneous state, and its molecular weight was found to be 106 kDa. It was composed of glucosamine, rhamnose, and galactose at a molar ratio of 86.4:1.6:1.0, respectively, indicating a rarely found novel high-glucosamine-containing biopolymer. The FT-IR and $^{13}C-NMR$ spectra of the novel cationic polyglucosamine biopolymer PGB-l revealed a close identity with chitosan from crab shell. It can effectively flocculate various suspended solids, including kaolin clay, $Ca(OH)_2,\;Al_{2}O_3$, active carbon, microbial cells, and acidic dyes.

A Generalized Calorie Estimation Algorithm Using 3-Axis Accelerometer

  • Choi, Jee-Hyun;Lee, Jeong-Whan;Shin, Kun-Soo
    • Journal of Biomedical Engineering Research
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    • v.27 no.6
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    • pp.301-309
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    • 2006
  • The main purpose of this study is to derive a regression equation that predicts the individual differences in activity energy expenditure (AEE) using accelerometer during different types of activity. Two subject groups were recruited separately in time: One is a homogeneous group of 94 healthy young adults with age ranged from $20\sim35$ yrs. The other subject group has a broad spectrum of physical characteristics in terms of age and fat ratio. 226 adolescents and adults of age ranged from $12\sim57$ yrs and fat ratio from $4.1\sim39.7%$ were in the second group. The wireless 3-axis accelerometers were developed and carefully fixed at the waist belt level. Simultaneously the total calorie expenditure was measured by gas analyzer. Each subject performed walking and running at speeds of 1.5, 3.0, 4.5, 6.0, 6.5, 7.5, and 8.5 km/hr. A generalized sensor-independent regression equation for AEE was derived. The regression equation was developed fur walking and running. The regression coefficients were predicted as functions of physical factors-age, gender, height, and weight with multivariable regression analysis. The generalized calorie estimation equation predicts AEE with correlation coefficient of 0.96 and the average accuracy of the accumulated calorie was $89.6{\pm}7.9%$.

A Study on the Wear Characteristics of SiC Particle Dispersed Composites by Rheo-Compocasting Method (Rheo-compocasting법에 의한 SiC입자분산 복합재료의 마모특성에 관한 연구)

  • Kwak, Hyun-Man;Choi, Chang-Ock
    • Journal of Korea Foundry Society
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    • v.13 no.3
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    • pp.238-247
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    • 1993
  • Microstructure, hardness and wear characteristics of $SiC_p/Al-6.5wt%Si-1.7wt%Mg$ alloy composites fabricated by the method of rheo-compocasting and hot pressing are investigated in this study. The dispersion of SiC particles in the composites is homogeneous and the hardness improves as additional amount increases. The wear amount of the matrix metal increases highly as wear rates increase, for the wear mechanism changes from adhesive wear to melt wear, and the matrix metal was coated on the surface of revolving disc and its weight increases. In the 5vol% composites, Fe is adhered on the surface of specimen by the projection of the dispersed hard SiC particles which have net-work structure and the coating layer is about $300{\mu}m$. But in the composite more than 20vol%, the wear amount of composite decreases because the SiC particles which have superior hardness, wear resistance and heat resistance properties resist wear, the abrasive wear turn out predominant wear mechanism and so the wear amount of revolving disc increases.

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Physical Properties of Hybrid Poplar Flakeboard Bonded with Alkaline Phenolic Soy Adhesives

  • Yang, In;Kuo, Monlin;Myers, Deland J.
    • Journal of the Korean Wood Science and Technology
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    • v.33 no.5 s.133
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    • pp.66-75
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    • 2005
  • Soybean-based adhesives have recently been reconsidered as alternatives to petroleum-based adhesives due to the uncertainty of availability of petrochemical products and the increased demand for wood adhesives. This study was conducted to investigate the adhesive properties of alkaline phenolic soy (APS) resin for hybrid poplar flakeboard. The APS resin was formulated by crosslinking an alkaline soy flour hydrolyzate with lab-prepared PF resin in the soy hydrolyzate to PF resin weight ratios of 70/30, 60/40, and 50/50. The APS resins were used to fabricate homogeneous hybrid poplar flakeboards with different resin solid levels (5%, 7%, and 9%), press temperatures (175 and $200^{\circ}C$), and press times of 8 and 10 minutes. The IB, wet MOR, and dimensional stability properties of board improved with increasing press time, press temperature, and PF level in APS resins. Increasing press time can be used to offset poor IB strength associated with a 9% resin solid level and the excessive moisture content in the mat. The following conditions were concluded to meet the requirements of the CSA standard for exterior-grade flakeboard: a 50% PF level, a 5% resin content, a $200^{\circ}C$ press temperature, and an 8 minute press time.

Catastrophe analysis of active-passive mechanisms for shallow tunnels with settlement

  • Yang, X.L.;Wang, H.Y.
    • Geomechanics and Engineering
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    • v.15 no.1
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    • pp.621-630
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    • 2018
  • In the note a comprehensive and optimal passive-active mode for describing the limit failure of circular shallow tunnel with settlement is put forward to predict the catastrophic stability during the geotechnical construction. Since the surrounding soil mass around tunnel roof is not homogeneous, with tools of variation calculus, several different curve functions which depict several failure shapes in different soil layers are obtained using virtual work formulae. By making reference to the simple-form of Power-law failure criteria based on numerous experiments, a numerical procedure with consideration of combination of upper bound theorem and stochastic medium theory is applied to the optimal analysis of shallow-buried tunnel failure. With help of functional catastrophe theory, this work presented a more accurate and optimal failure profile compared with previous work. Lastly the note discusses different effects of parameters in new yield rule and soil mechanical coefficients on failure mechanisms. The scope of failure block becomes smaller with increase of the parameter A and the range of failure soil mass tends to decrease with decrease of unit weight of the soil and tunnel radius, which verifies the geomechanics and practical case in engineering.

Purification and Biophysicochemical Properties of the Virus Causing Necrotic Spot Symptom on Korean Cactus (한국산 접목 선인장에서 분리한 괴사 반점 원인 바이러스의 생물리화학적 특성)

  • 이철호;나용준;한정헌
    • Korean Journal Plant Pathology
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    • v.13 no.2
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    • pp.73-78
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    • 1997
  • A virus was purified from Korean cactus (Chamaeaereus silverstrii f. Variegata Hort) showing necrotic spots and its biophysicochemical properties were examined. The typical necrotic spots were observed on healthy looking cactus inoculated with the purified virus and the various symptoms from necrotic spots to systemic were observed on 17 species of test plats used, which suggests that the purified virus has the normal infectivity and wide host range. Ultraviolet absorption scanning analysis revealed that the viral preparation had the typical curve with shoulder at 290 nm and its ratio of absorbances at 260 nm to 280 nm and extinction coefficient was 1.156 and 2.86, respectively. The virus represented one distinct and homogeneous band and molecular weight of viral coat protein was determined to 17, 000 Dalton by SDS-PAGE. The virus revealed rod-shaped virion (325$\times$20 nm) by electron microscopy. Based on these results, it has been thought that the virus purified in this study is the pathogenic agents causing necrotic spot symptom on Korean cacuts and one of the member of tobamovirus family.

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