• Title/Summary/Keyword: Control Derivatives

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Comparisons of Recombinant Protein Expression in Diverse Natural Isolates of Escherichia coli

  • Jung, Yuna;Lim, Dongbin
    • Molecules and Cells
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    • v.25 no.3
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    • pp.446-451
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    • 2008
  • We assessed heterologous protein expression in 64 strains obtained from the Escherichia coli Reference (ECOR) collection, a collection representing diverse natural E. coli populations. A plasmid generating a glutathione S-transferase and plant carbonic anhydrase fusion protein (GST-CA) under the control of the tac promoter was introduced into the ECOR strains, and the quantity of the fusion protein was determined by SDS-PAGE. The foreign protein was generated at various levels, from very high (40 strains, high producers) to very low (six strains, low producers). Immunoblotting showed that the high producers expressed approximately 250-500 times more GST-CA protein than the low producers. The results of semi-quantitative RT-PCR showed that the low producers generated mRNA levels comparable to those of the high producers, thereby suggesting that, at least in this case, inefficient translation is a major cause of the low production. We introduced a different plasmid, which expressed a maltose binding protein and plant guanylate kinase fusion protein (MBP-GK) into the six low producers. Interestingly, five of these expressed MBP-GK at very high levels. Thus, we conclude that the production of a particular protein from an expression vector can vary considerably, depending on the host strain. Strains in the ECOR collection could function as useful alternative hosts when a desired level of protein expression is not obtained from commonly used strains, such as E. coli K12 or B derivatives.

Effect of Cell Aggregation on Camptothecin Production in Suspension Cultures of Camptotheca acuminata (Camptotheca acuminata 현탁배양에서 세포응집이 Camptothecin 생산에 미치는 영향)

  • Choi, Hoon;Byun, Sang-Yo
    • KSBB Journal
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    • v.15 no.6
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    • pp.609-614
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    • 2000
  • Suspension cultures of Camptotheca acuminata, which is known to produce the anticancer indole alkaloid camptothecin and its derivatives, were made to increase camptothecin production. The capability of camptothecin production in suspended cells is decreased by repeated subculturein. Aggregated cells produced more camptothecin than single cells. Optimal cell aggregation was achieved in hybrid medium supplemented with 4% sucrose. Aggregated cells in hybrid medium with 4% sucrose produced $18.04{\times}10^{-4} mg/L$ of camptothecin. The control of shaking speeds was effective at inducing cell aggregation and camptothecin production. A shaking speed of 100 rpm was found optimum to increase the cell aggregation with a camptothecin production of $19.4{\times}10^{-4} mg/L$.

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Phosphoinositide turnover in Toll-like receptor signaling and trafficking

  • Le, Oanh Thi Tu;Nguyen, Tu Thi Ngoc;Lee, Sang Yoon
    • BMB Reports
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    • v.47 no.7
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    • pp.361-368
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    • 2014
  • Lipid components in biological membranes are essential for maintaining cellular function. Phosphoinositides, the phosphorylated derivatives of phosphatidylinositol (PI), regulate many critical cell processes involving membrane signaling, trafficking, and reorganization. Multiple metabolic pathways including phosphoinositide kinases and phosphatases and phospholipases tightly control spatio-temporal concentration of membrane phosphoinositides. Metabolizing enzymes responsible for PI 4,5-bisphosphate (PI(4,5)P2) production or degradation play a regulatory role in Toll-like receptor (TLR) signaling and trafficking. These enzymes include PI 4-phosphate 5-kinase, phosphatase and tensin homolog, PI 3-kinase, and phospholipase C. PI(4,5)P2 mediates the interaction with target cytosolic proteins to induce their membrane translocation, regulate vesicular trafficking, and serve as a precursor for other signaling lipids. TLR activation is important for the innate immune response and is implicated in diverse pathophysiological disorders. TLR signaling is controlled by specific interactions with distinct signaling and sorting adaptors. Importantly, TLR signaling machinery is differentially formed depending on a specific membrane compartment during signaling cascades. Although detailed mechanisms remain to be fully clarified, phosphoinositide metabolism is promising for a better understanding of such spatio-temporal regulation of TLR signaling and trafficking.

Inhibitors of Nitric Oxide Syntheasis from Phellinus pini in Murine Macrophages (낙엽송층버섯의 Nitric Oxide 생성저해 물질)

  • Jang, Hyun-Jin;Kim, Ahn-Keun;Pyo, Myoung-Yun;Yang, Ki-Sook
    • YAKHAK HOEJI
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    • v.51 no.6
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    • pp.430-434
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    • 2007
  • The anti-inflammatory activity of fruit body of Phellinus pini was investigated by activity-guided fractionation. From the screening of each fraction for the inhibitory activity of NO production in lipopolysaccaride (LPS) activated RAW 264.7 cells, methanol extract and its hexane soluble fraction of Phellinus pini exhibited inhibition of NO production compared with LPS control without toxicity. The hexane soluble fraction showed dose-dependent inhibition of NO production. The active hexane fraction was repeatedly chromatographed over silica gel, ergosta-7,24(28)-dien-3-ol(1) and ergosterol peroxide (2) were isolated and identified. Ergosterol derivatives were inhibited NOS activation, $IC_{50}$ of them were $18.9{\pm}3.9{\mu}M$ (1) and $20.4{\pm}4.5{\mu}M$ (2).

Protective Effects of Thiazolo[3,2-b]-1,2,4-Triazoles on Ethanol­Induced Oxidative Stress in Mouse Brain and Liver

  • Aktay Goknur;Tozkoparan Birsen;Ertan Mevlut
    • Archives of Pharmacal Research
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    • v.28 no.4
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    • pp.438-442
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    • 2005
  • A series of 3-[1-(4-(2-methylpropyl) phenyl) ethyl]-1,2,4-triazole-5-thione (I) and its bicyclic condensed derivatives 6-benzylidenethiazolo[3,2-b]-1, 2,4-triazole-5(6H)-ones (IIa-IIf) were investigated for the prevention of ethanol-induced oxidative stress in liver and brain of mice. Administration of ethanol (0.1 mL/mice, p.o.) resulted in a drop of total thiol groups (T-SH) and non-protein thiol groups (NP-SH), and an increase in thiobarbituric acid reactive substances (TBARS) in both liver and brain tissue of mice (p<0.001). Among the compounds investigated (at a dose of 200 mg/kg, p.o.), I and IId ameliorated the peroxidative injury in these tissues effectively. Compounds IIa, IIc and IIe improved the peroxidative tissue injury only in brain. These findings suggest that certain condensed thiazolo-triazole compounds may contribute to the control of ethanol-induced oxidative stress in an organ selective manner.

Conjugated Linoleic Acid Changes fatty Acid Composition by Decreasing Monounsaturated fatty Acids in Rabbits and Hep G2 Cells

  • Nam, Kisun
    • Journal of Nutrition and Health
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    • v.30 no.4
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    • pp.442-450
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    • 1997
  • Conjugated dienoic derivatives of linoleic acid(CLA) are a mixture of positional and geometric isomers of linoleic acid(LA). We previously found that CLA changes the fatty acid profile in chicken eggs and serum by decreasing monounsaturated fatty acids. Studies were conducted to explore the effects of CLA on fatty acid composition. Rabbits were fed a semisynthetic diet with or without CLA(0.5g CLA/rabbit/day) for 22 weeks. Compared to the control, rabbits fed CLA had significantly lower monounsaturated fatty acid levels(palmitoleic acid Cl6 : 1 by 50% and oleic acid Cl8 : 1, by 20%) in plasma lipids. We found similar differences in fatty acid composition in the liver and the aorta. The inhibitory effect of CLA on $\Delta$9 desaturation was confirmed in a human hepatoma cell line, Hep G2. CLA significantly decreased $\Delta$9 desaturation in 4-5 hours as shown by an increase in the ratio of Cl6 : 0 to C 16 1, This is apparently due to a decrease in $\Delta$9 desaturase(stearoyl-CoA desaturase, SCD) activity ; it was decreased more than 50%. These results, along with our previous findings, indicate that CLA is an inhibitor of $\Delta$9 desaturase in the liver.

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Isogeometric Collocation Method to solve the strong form equation of UI-RM Plate Theory

  • Katili, Irwan;Aristio, Ricky;Setyanto, Samuel Budhi
    • Structural Engineering and Mechanics
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    • v.76 no.4
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    • pp.435-449
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    • 2020
  • This work presents the formulation of the isogeometric collocation method to solve the strong form equation of a unified and integrated approach of Reissner Mindlin plate theory (UI-RM). In this plate theory model, the total displacement is expressed in terms of bending and shear displacements. Rotations, curvatures, and shear strains are represented as the first, the second, and the third derivatives of the bending displacement, respectively. The proposed formulation is free from shear locking in the Kirchhoff limit and is equally applicable to thin and thick plates. The displacement field is approximated using the B-splines functions, and the strong form equation of the fourth-order is solved using the collocation approach. The convergence properties and accuracy are demonstrated with square plate problems of thin and thick plates with different boundary conditions. Two approaches are used for convergence tests, e.g., increasing the polynomial degree (NELT = 1×1 with p = 4, 5, 6, 7) and increasing the number of element (NELT = 1×1, 2×2, 3×3, 4×4 with p = 4) with the number of control variable (NCV) is used as a comparable equivalent variable. Compared with DKMQ element of a 64×64 mesh as the reference for all L/h, the problem analysis with isogeometric collocation on UI-RM plate theory exhibits satisfying results.

Fabrication and Evaluation of Polyelectrolyte Complexes of Dextran Derivatives for Drug Coating of Coronary Stents

  • Jang, Eun-Jae;Lee, So-Youn;Bae, In-Ho;Park, Dae Sung;Jeong, Myung Ho;Park, Jun-Kyu
    • Applied Chemistry for Engineering
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    • v.30 no.5
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    • pp.586-590
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    • 2019
  • The aim of this study was to fabricate a dextran polyelectrolyte multi-layer on a bare metal stent (BMS) and to evaluate bio-physical properties of the layer. Diethylaminoethyl-dextran (DEAE-D) as a polycation and dextran sulfate (DS) as a polyanion were successively coated on the bare metal stent by a well-known layer-by-layer procedure. The morphology of the stent surface and its cell adhesion were studied after each coating step by scanning electron microscopy. The stent showed more blotched and slightly rougher morphology after dextran-DS coating. The contact angle of the DEAE-DS group ($39.5{\pm}0.15^{\circ}$) was significantly higher than that of the BMS group ($45.16{\pm}0.08^{\circ}$), indicating the improvement of hydrophilic. The SMC proliferation inhibition in the DEAE-DS-coated stent group ($20.9{\pm}0.04%$) was stronger than that in the control group ($21.7{\pm}0.10%$ in DS-coated group only). The DEAE-DS coating is desired for stent coating materials with biocompatibility and anti-restenosis effect.

Microwave Assisted Synthesis of Graphene-Bi2MoO6 Nanocomposite as Sono-Photocatalyst

  • Tang, Jia-Yao;Zhu, Lei;Fan, Jia-Yi;Sun, Chen;Oh, Won-Chun
    • Korean Journal of Materials Research
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    • v.32 no.1
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    • pp.1-8
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    • 2022
  • In this investigation, Bi2MoO6 deposited graphene nanocomposite (BMG) was synthesized using a simple microwave assisted hydrothermal synthesis method. The synthesized BMG nanocomposite was characterized by X-ray diffraction, transmission electron microscopy, scanning electron microscopy with energy dispersive X-ray analysis, and photocurrent analysis. The study revealed that the catalysts prepared have high crystalline nature, enhanced light responsive property, high catalytic activity, and good stability. XRD results of BMG composite exhibit a koechlinite phase of Bi2MoO6. The surface property is shown by SEM and TEM, which confirmed a homogenous composition in the bulk particles of Bi2MoO6 and nanosheets of graphene. The catalytic behavior was investigated by the decomposition of Rhodamine B as a standard dye. The results exhibit excellent yields of product derivatives at mild conditions under ultrasonic/visible light-medium. Approximately 1.6-times-enhanced sono-photocatalytic activity was observed by introduction of Bi2MoO6 on graphene nanosheet compared with control sample P25 during 50 min test.

Useful Applications Decursinol Derivatives as Biochemical Fungicides (데커시놀 파생물질들의 항징균제로서의 활용)

  • Yun-Jeong Shin;Jiyoung Shin;Hyewon Jang;Hokyoung Son;Yonghoon Kwon
    • Proceedings of the Plant Resources Society of Korea Conference
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    • 2022.09a
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    • pp.30-30
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    • 2022
  • Natural products decursin and decursinol angelate were recently reported as benign fungicides for controlling rice blast. Inspired by the structural similarity of the cumarin compounds and gained hint from the skeletal motifs, we designed and prepared synthetic compounds to increase the natural product efficacy and evaluated their antifungal activities against various plant disease pathogens in vitro. Synthetically prepared compound 4 and 5 indeed suppressed the mycelial growth of B. cinerea, F. oxysporum, P. italicum, and R. quercus-mongolicae. Additionally, compound 5 effectively prevents the growth of C. coccodes and C. parasitica. Furthermore, both 4 and 5 possess better inhibitory activities on spore germination of F. oxysporum and M. oryzae than the natural product decursin. These results suggest that the effect of the lead compound for plant disease protection can be improved by tuning the structure of the original natural product and decursinol chloroacrylates 4 and 5 are candidates for the control of F. oxysporum and M. oryzae.

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