• Title/Summary/Keyword: Activity state

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An inhibitory role of NEK6 in TGFβ/Smad signaling pathway

  • Zuo, Jie;Ma, Haijie;Cai, Hao;Wu, Yanhua;Jiang, Wei;Yu, Long
    • BMB Reports
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    • v.48 no.8
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    • pp.473-478
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    • 2015
  • The NEK6 (NIMA-related kinases 6) is reported to play po-tential roles in tumorigenesis. Although it is suggested to function in several cellular pathways, the underlying mechanism in tumorigenesis is still largely unknown. In the present study, we discovered interaction of NEK6 with Smad4, a key member of transforming growth factor beta (TGFβ) pathway. Over-expression of NEK6 in hepatocellular carcinoma (HCC) cell lines suppresses TGFβ-mediated transcription activity in a kinase activity-dependent manner. In addition, NEK6 suppresses the cell growth arrest induced by TGFβ. Mechanically, NEK6 blocks nuclear translocation of Smad4, which is essential for TGFβ function. Moreover, we identified that NEK6 could be regulated by TGFβ and hypoxia. Our study sheds new light on the roles of NEK6 in canonical TGFβ/Smad pathway and tum-origenesis. [BMB Reports 2015; 48(8): 473-478]

Pedagogical Analysis Of The Phenomenon Of Digital Competence

  • Yovenko, Larysa;Novakivska, Lyudmyla;Sanivskyi, Oleksandr;Sherman, Mykhailo;Vysochan, Lesia;Hnedko, Natalia
    • International Journal of Computer Science & Network Security
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    • v.21 no.6
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    • pp.7-10
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    • 2021
  • The article analyzes and concretizes the understanding of the differences between the concepts of competence / competence according to the criterion general - personal. Based on the identified characteristics of competence (completed personal quality, activity character, educational result, successful implementation of professional and educational activities), the concept of competence as an integrative dynamic quality of a person, manifested in effective activity in a specific area, is defined. The structure of the IC has been substantiated, including motivational and value; information technology; communicative and reflective components. The content of the named IC components is disclosed. The article analyzes the essence of the characteristics of basic concepts (competence / competence), consideration of information competence in the research of famous scientists in order to concretize the studied phenomenon; concretization of the identified pedagogical conditions in educational process.

Different Functional and Structural Characteristics between ApoA-I and ApoA-4 in Lipid-Free and Reconstituted HDL State: ApoA-4 Showed Less Anti-Atherogenic Activity

  • Yoo, Jeong-Ah;Lee, Eun-Young;Park, Ji Yoon;Lee, Seung-Taek;Ham, Sihyun;Cho, Kyung-Hyun
    • Molecules and Cells
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    • v.38 no.6
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    • pp.573-579
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    • 2015
  • Apolipoprotein A-I and A-IV are protein constituents of high-density lipoproteins although their functional difference in lipoprotein metabolism is still unclear. To compare anti-atherogenic properties between apoA-I and apoA-4, we characterized both proteins in lipid-free and lipidbound state. In lipid-free state, apoA4 showed two distinct bands, around 78 and $67{\AA}$ on native gel electrophoresis, while apoA-I showed scattered band pattern less than $71{\AA}$. In reconstituted HDL (rHDL) state, apoA-4 showed three major bands around $101{\AA}$ and $113{\AA}$, while apoA-I-rHDL showed almost single band around $98{\AA}$ size. Lipid-free apoA-I showed 2.9-fold higher phospholipid binding ability than apoA-4. In lipid-free state, $BS_3$-crosslinking revealed that apoA-4 showed less multimerization tendency upto dimer, while apoA-I showed pentamerization. In rHDL state (95:1), apoA-4 was existed as dimer as like as apoA-I. With higher phospholipid content (255:1), five apoA-I and three apoA-4 were required to the bigger rHDL formation. Regardless of particle size, apoA-I-rHDL showed superior LCAT activation ability than apoA-4-rHDL. Uptake of acetylated LDL was inhibited by apoA-I in both lipid-free and lipid-bound state, while apoA-4 inhibited it only lipid-free state. ApoA-4 showed less anti-atherogenic activity with more sensitivity to glycation. In conclusion, apoA-4 showed inferior physiological functions in lipid-bound state, compared with those of apoA-I, to induce more pro-atherosclerotic properties.

Butyrolactones Derivatives from the Fermentation Products of an Endophytic Fungus Aspergillus versicolor

  • Ye, Yan-Qing;Xia, Cong-Fang;Yang, Juan-Xia;Yang, Yu-Chun;Qin, Ying;Gao, Xue-Mei;Du, Gang;Li, Xue-Mei;Hu, Qiu-Fen
    • Bulletin of the Korean Chemical Society
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    • v.35 no.10
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    • pp.3059-3062
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    • 2014
  • Two new butyrolactones, asperphenol A (1) and B (2), together with four known butyrolactones (3-6) were isolated from the fermentation products of an endophytic fungus Aspergillus versicolor. Their structures were elucidated by spectroscopic methods including extensive 1D- and 2D-NMR techniques. Compounds 1-6 were also tested for their anti-tobacco mosaic virus (anti-TMV) activities. The results showed that compound 2 exhibited high anti-TMV activity with inhibition rate of 46.7%. The other compounds also exhibited potential anti-TMV activities with inhibition rates in the range of 21.8-28.4%.

Cotton GhKCH2, a Plant-specific Kinesin, is Low-affinitive and Nucleotide-independent as Binding to Microtubule

  • Xu, Tao;Sun, Xuewei;Jiang, Shiling;Ren, Dongtao;Liu, Guoqin
    • BMB Reports
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    • v.40 no.5
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    • pp.723-730
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    • 2007
  • Kinesin is an ATP-driven microtubule motor protein that plays important roles in control of microtubule dynamics, intracellular transport, cell division and signal transduction. The kinesin superfamily is composed of numerous members that are classified into 14 subfamilies. Animal kinesins have been well characterized. In contrast, plant kinesins have not yet to be characterized adequately. Here, a novel plant-specific kinesin gene, GhKCH2, has been cloned from cotton (Gossypium hirsutum) fibers and biochemically identified by prokaryotic expression, affinity purification, ATPase activity assay and microtubule-binding analysis. The putative motor domain of GhKCH2, $M_{396-734}$ corresponding to amino acids Q396-N734 was fused with 6$\times$His-tag, soluble-expressed in E. coli and affinity-purified in a large amount. The biochemical analysis demonstrated that the basal ATPase activity of $M_{396-734}$ is not activated by $Ca^{2+}$, but stimulated 30-fold max by microtubules. The enzymatic activation is microtubule-concentration-dependent, and the concentration of microtubules that corresponds to half-maximum activation was about 11 ${\mu}M$, much higher than that of other kinesins reported. The cosedimentation assay indicated that $M_{396-734}$ could bind to microtubules in vitro whenever the nucleotide AMP-PNP is present or absent. As a plant-specific microtubule-dependent kinesin with a lower microtubule-affinity and a nucleotide-independent microtubule-binding ability, cotton GhKCH2 might be involved in the function of microtubules during the deposition of cellulose microfibrils in fibers or the formation of cell wall.

Biochemical Characterization of Exoribonuclease Encoded by SARS Coronavirus

  • Chen, Ping;Jiang, Miao;Hu, Tao;Liu, Qingzhen;Chen, Xiaojiang S.;Guo, Deyin
    • BMB Reports
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    • v.40 no.5
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    • pp.649-655
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    • 2007
  • The nsp14 protein is an exoribonuclease that is encoded by severe acute respiratory syndrome coronavirus (SARS-CoV). We have cloned and expressed the nsp14 protein in Escherichia coli, and characterized the nature and the role(s) of the metal ions in the reaction chemistry. The purified recombinant nsp14 protein digested a 5'-labeled RNA molecule, but failed to digest the RNA substrate that is modified with fluorescein group at the 3'-hydroxyl group, suggesting a 3'-to-5' exoribonuclease activity. The exoribonuclease activity requires $Mg^{2+}$ as a cofactor. Isothermal titration calorimetry (ITC) analysis indicated a two-metal binding mode for divalent cations by nsp14. Endogenous tryptophan fluorescence and circular dichroism (CD) spectra measurements showed that there was a structural change of nsp14 when binding with metal ions. We propose that the conformational change induced by metal ions may be a prerequisite for catalytic activity by correctly positioning the side chains of the residues located in the active site of the enzyme.

Cloning, Expression, and Characterization of a Highly Active Alkaline Pectate Lyase from Alkaliphilic Bacillus sp. N16-5

  • Li, Gang;Rao, Lang;Xue, Yanfen;Zhou, Cheng;Zhang, Yun;Ma, Yanhe
    • Journal of Microbiology and Biotechnology
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    • v.20 no.4
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    • pp.670-677
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    • 2010
  • An alkaline pectate lyase, Bsp165PelA, was purified to homogeneity from the culture broth of alkaliphilic Bacillus sp. N16-5. The enzyme showed a specific activity as high as 1,000 U/mg and had optimum activity at pH 11.5 and $50^{\circ}C$. It was composed of a single polypeptide chain with a molecular mass of 42 kDa deduced from SDS-PAGE, and its isoelectric point was around pH 6.0. It could efficiently depolymerize polygalacturonate and pectin. Characterization of product formation revealed unsaturated digalacturonate and trigalacturonate as the main products. The pectate lyase gene (pelA) contained an open reading frame (ORF) of 1,089 bp, encoding a 36-amino acids signal peptide and a mature protein of 326 amino acids with a calculated molecular mass of 35.943 Da. The deduced amino acid sequence from the pelA ORF exhibited significant homology to those of known pectate lyases in polysaccharide lyase family 1. Some conserved active-site amino acids were found in the deduced amino acid sequence of Bsp165PelA. $Ca^{2+}$ was not required for activity on pectic substrates.

A Novel pH-Stable, Bifunctional Xylanase Isolated from a Deep-Sea Microorganism, Demequina sp. JK4

  • Meng, Xin;Shao, Zongze;Hong, Yuzhi;Lin, Ling;Li, Chanjuan;Liu, Ziduo
    • Journal of Microbiology and Biotechnology
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    • v.19 no.10
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    • pp.1077-1084
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    • 2009
  • A genomic library was constructed to clone a xylanase gene (Mxyn10) from Demequina sp. JK4 isolated from a deep sea. Mxyn10 encoded a 471 residue protein with a calculated molecular mass of 49 kDa. This protein showed the highest sequence identity (70%) with the xylanase from Streptomyces lividans. Mxyn10 contains a catalytic domain that belongs to the glycoside hydrolase family 10 (GH10) and a carbohydrate-binding module (CBM) belonging to family 2. The optimum pH and temperature for enzymatic activity were pH 5.5 and $55^{\circ}C$, respectively. Mxyn10 exhibited good pH stability, remaining stable after treatment with buffers ranging from pH 3.5 to 10.0. The protein was not significantly affected by a variety of chemical reagents, including some compounds that usually inhibit the activity of other related enzymes. In addition, Mxyn10 showed activity on cellulose. These properties mark Mxyn10 as a potential enzyme for industrial application and saccharification processes essential for bioethanol production.

Quitline Activity in Rajasthan, India

  • Gupta, Rakesh;Verma, Vinit;Mathur, Pankaj
    • Asian Pacific Journal of Cancer Prevention
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    • v.17 no.sup2
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    • pp.19-24
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    • 2016
  • Quitline activity in Rajasthan, India is a voluntary activity of Rajasthan Cancer Foundation (RCF) since April 2013. To kick-off, it took the benefit of the State Government- PIRAMAL SWASTHYA (PS)1 collaborative 104 Health Information Helpline that existed already in public-private partnership. It is a reactive quitline that helps callers through the counselors and nursing staff trained specifically through the weekly sessions held by the first author, the RCF resource on quitline. Besides structuring of the scripts for primary intervention and follow-ups after 1 week, 1 month, 6 months and a year, he also monitors calls, advices and coordinates with the supervisors to manage and analyze the data base, and reports to the PS lead at the Jaipur Center on overall performance and to plan strategic communication with the State Government on its outcomes. The quitline has limitations of its informal existence through a voluntary effort of RCF, no specific resource allocation, suboptimal data management, minimal awareness in the masses due to poor IEC (Information, Education and Communication; except its efforts made by RCF in last 1 year through the government-run State TV and City Radio) and staff shortage and its attrition due to lack of plan for career advancement. Despite these challenges in the year 2013, the quit line has registered a quit rate (for complete abstinence) of 19.93% amongst 1525 callers. The quit rate were 58.01% (304/ 524) among the responders at the 3rd follow-up at 18 months (in September 2014)2. In view of an increase in quit rate by 5- 9 times over the prevailing quit rate in the former ever daily users [both smokers and the users of smokeless tobacco (SLT)], efforts are being made by RCF in concurrence with PS to have this cost-effective model established formally with optimal resource allocation in collaboration with willing agencies (the State and Central Governments and the International Quitline Agencies) and its replication in 4 more states where PS is collaborating with the respective state governments similarly (Assam, Chhattisgarh, Jharkhand and Karnataka).

Cross-talk between STAT6 and Ras/MAPK Pathway for the IL-4-mediated T Cell Survival

  • So, Eui-Young;Jang, Ji-Young;Lee, Choong-Eun
    • BMB Reports
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    • v.34 no.6
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    • pp.578-583
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    • 2001
  • As a prototypic Thl vs Th2 cytokine, IFN-$\gamma$ and IL-4 activate distinct STAT proteins, STAT1 and STATE, respectively. In cytokine-producing Jurkat T cells, IL-4 is effectively rescued from cell death that is induced by dexamethasone, but IFN-$\gamma$ failed to do so. Since the Ras/MAPK pathway is known to play an important role in cytokine-induced cell survival, we investigated the mechanism of T cell survival through the analysis of functional cross-talk between Ras/MAPK and distinct STAT proteins that are activated by IL-4 and IFN-$\gamma$. Although IL-4 and IFN-$\gamma$ each induced the activation of STATE and STATI. in Jurkat T cells, respectively, only IL-4 was capable of inducing MAPK. Along with tyrosine kinase inhibitors, MEK/MAPK inhibitors also caused a significant suppression of the IL-4-induced STATE activity. This suggests a positive regulation of STATE by MAPK during IL-4 signal transduction. Furthermore, transfection studies with dominant active (da) vs dominant negative (dn) Ras revealed that daRas, but not dnRas, selectively up-regulated the expression and activity of STATE with a concomitant increase in MAPK activity. These results, therefore, suggest that there is a functional cross-talk between the Ras/MAPK and Jak/STAT6 pathways, which may have a role in the IL-4-induced T cell survival.

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