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Sample records for high constitutive expression

  1. Isolation and analysis of two Escherichia coli K-12 ilv attenuator deletion mutants with high-level constitutive expression of an ilv-lac fusion operon.

    OpenAIRE

    Bennett, D. C.; Umbarger, H E

    1984-01-01

    A lysogenizing lambda phage, lambda dilv-lac11, was constructed to carry an ilvD-lac operon fusion. Expression from the phage of the ilvE and lacZ genes is controlled by an intact ilv control region also carried by this phage. Two spontaneous mutants of lambda dilv-lac11 that have high-level constitutive expression of the ilv-lac fusion operon were isolated by growth on a beta-chloroalanine selective medium. The mutants were shown by nucleotide sequence determination to contain large deletion...

  2. A novel PR10 promoter from Erianthus arundinaceus directs high constitutive transgene expression and is enhanced upon wounding in heterologous plant systems.

    Science.gov (United States)

    Chakravarthi, M; Syamaladevi, Divya P; Harunipriya, P; Augustine, Sruthy Maria; Subramonian, N

    2016-01-01

    In genetic engineering, inducible promoters play an important role as the expression of genes driven by them can be turned on or off under situations like biotic or abiotic factors. There are few reports on inducible promoters that can be employed in the development of transgenic plants, particularly in sugarcane. In the present study, four wound inducible genes (Chitinase, PR1A, PR10 and HRGP) were selected and were amplified from Erianthus arundinaceus, a distant relative of sugarcane. In order to determine the gene that is highly induced upon wounding, RT-qPCR was performed, which showed that PR10 gene expression was instantaneous and higher upon wounding when compared to the other three genes. Using the random amplification of genomic ends technique, a 592 bp promoter sequence was obtained and in silico analysis of the upstream regulatory region revealed a 469 bp promoter and 123 bp of 5' untranslated region (UTR). Functional analyses of the promoter sequence (with and without 5' UTR) in tobacco, rice and sugarcane using β-glucuronidase (GUS) as the reporter gene revealed the constitutive and inducible nature of the PR10 promoter. Our studies have demonstrated that the PR10 promoter, though highly constitutive, was quickly induced upon wounding as well as on treatment with abscisic acid and methyl jasmonate hormones. This is the first report on the isolation and characterization of a PR10 promoter from a wild grass and is expected to have application for development of transgenic plants.

  3. Constitutive high expression of chromosomal beta-lactamase in Pseudomonas aeruginosa caused by a new insertion sequence (IS1669) located in ampD

    DEFF Research Database (Denmark)

    Bagge, Niels; Ciofu, Oana; Hentzer, Morten

    2002-01-01

    The expression of chromosomal AmpC beta-lactamase in Pseudomonas aeruginosa is negatively regulated by the activity of an amidase, AmpD. In the present study we examined resistant clinical P. aeruginosa strains and several resistant variants isolated from in vivo and in vitro biofilms for mutations...... in the expression of high levels of AmpC beta-lactamase. Complementation of these isolates with ampD from the reference P. aeruginosa strain PAO1 caused a dramatic decrease in the expression of AmpC beta-lactamase and a parallel decrease of the MIC of ceftazidime to a level comparable to that of PAO1. One highly...... resistant, constitutive beta-lactamase-producing variant contained no mutations in ampD, but a point mutation was observed in ampR, resulting in an Asp-135-->Asn change. An identical mutation of AmpR in Enterobacter cloacae has been reported to cause a 450-fold higher AmpC expression. However, in many...

  4. Constitutive high expression of chromosomal beta-lactamase in Pseudomonas aeruginosa caused by a new insertion sequence (IS1669) located in ampD

    DEFF Research Database (Denmark)

    Bagge, N.; Ciofu, O.; Hentzer, Morten;

    2002-01-01

    resistant, constitutive beta-lactamase-producing variant contained no mutations in ampD, but a point mutation was observed in ampR, resulting in an Asp-135-->Asn change. An identical mutation of AmpR in Enterobacter cloacae has been reported to cause a 450-fold higher AmpC expression. However, in many...

  5. Constitutive high-level expression of a codon-optimized β-fructosidase gene from the hyperthermophile Thermotoga maritima in Pichia pastoris.

    Science.gov (United States)

    Menéndez, Carmen; Martínez, Duniesky; Trujillo, Luis E; Mazola, Yuliet; González, Ernesto; Pérez, Enrique R; Hernández, Lázaro

    2013-02-01

    Enzymes for use in the sugar industry are preferred to be thermotolerant. In this study, a synthetic codon-optimized gene encoding a highly thermostable β-fructosidase (BfrA, EC 3.2.1.26) from the bacterium Thermotoga maritima was expressed in the yeast Pichia pastoris. The gradual increase of the transgene dosage from one to four copies under the control of the constitutive glyceraldehyde 3-phosphate dehydrogenase promoter had an additive effect on BfrA yield without causing cell toxicity. Maximal values of cell biomass (115 g/l, dry weight) and overall invertase activity (241 U/ml) were reached at 72 h in fed-batch fermentations using cane sugar as the main carbon source for growth. Secretion driven by the Saccharomyces cerevisiae α-factor signal peptide resulted in periplasmic retention (44 %) and extracellular release (56 %) of BfrA. The presence of N-linked oligosaccharides did not influence the optimal activity, thermal stability, kinetic properties, substrate specificity, and exo-type action mode of the yeast-secreted BfrA in comparison to the native unglycosylated enzyme. Complete inversion of cane sugar at initial concentration of 60 % (w/v) was achieved by periplasmic BfrA in undisrupted cells reacting at pH 5.5 and 70 °C, with average productivity of 4.4 g of substrate hydrolyzed per grams of biomass (wet weight) per hour. The high yield of fully active glycosylated BfrA here attained by recombinant P. pastoris in a low-cost fermentation process appears to be attractive for the large-scale production of this thermostable enzyme useful for the manufacture of inverted sugar syrup.

  6. A novel bicistronic high-capacity gutless adenovirus vector that drives constitutive expression of herpes simplex virus type 1 thymidine kinase and tet-inducible expression of Flt3L for glioma therapeutics.

    Science.gov (United States)

    Puntel, Mariana; Muhammad, A K M G; Candolfi, Marianela; Salem, Alireza; Yagiz, Kader; Farrokhi, Catherine; Kroeger, Kurt M; Xiong, Weidong; Curtin, James F; Liu, Chunyan; Bondale, Niyati S; Lerner, Jonathan; Pechnick, Robert N; Palmer, Donna; Ng, Philip; Lowenstein, Pedro R; Castro, Maria G

    2010-06-01

    Glioblastoma multiforme (GBM) is a deadly primary brain tumor. Conditional cytotoxic/immune-stimulatory gene therapy (Ad-TK and Ad-Flt3L) elicits tumor regression and immunological memory in rodent GBM models. Since the majority of patients enrolled in clinical trials would exhibit adenovirus immunity, which could curtail transgene expression and therapeutic efficacy, we used high-capacity adenovirus vectors (HC-Ads) as a gene delivery platform. Herein, we describe for the first time a novel bicistronic HC-Ad driving constitutive expression of herpes simplex virus type 1 thymidine kinase (HSV1-TK) and inducible Tet-mediated expression of Flt3L within a single-vector platform. We achieved anti-GBM therapeutic efficacy with no overt toxicities using this bicistronic HC-Ad even in the presence of systemic Ad immunity. The bicistronic HC-Ad-TK/TetOn-Flt3L was delivered into intracranial gliomas in rats. Survival, vector biodistribution, neuropathology, systemic toxicity, and neurobehavioral deficits were assessed for up to 1 year posttreatment. Therapeutic efficacy was also assessed in animals preimmunized against Ads. We demonstrate therapeutic efficacy, with vector genomes being restricted to the brain injection site and an absence of overt toxicities. Importantly, antiadenoviral immunity did not inhibit therapeutic efficacy. These data represent the first report of a bicistronic vector platform driving the expression of two therapeutic transgenes, i.e., constitutive HSV1-TK and inducible Flt3L genes. Further, our data demonstrate no promoter interference and optimum gene delivery and expression from within this single-vector platform. Analysis of the efficacy, safety, and toxicity of this bicistronic HC-Ad vector in an animal model of GBM strongly supports further preclinical testing and downstream process development of HC-Ad-TK/TetOn-Flt3L for a future phase I clinical trial for GBM.

  7. GENERAL EXPRESSIONS OF CONSTITUTIVE EQUATIONS FOR ISOTROPIC ELASTIC DAMAGED MATERIALS

    Institute of Scientific and Technical Information of China (English)

    唐雪松; 蒋持平; 郑健龙

    2001-01-01

    The general expressions of constitutive equations for isotropic elastic damaged materials were derived directly from the basic law of irreversible thermodynamics. The limitations of the classical damage constitutive equation based on the well-known strain equivalence hypothesis were overcome. The relationships between the two elastic isotropic damage models(i. e. single and double scalar damage models)were revealed. When a single scalar damage variable defined according to the microscopic geometry of a damaged material is used to describle the isotropic damage state, the constitutive equations contain two "damage effect functions", which describe the different influences of damage on the two independent elastic constants. The classical damage constitutive equation based on the strain equivalence hypothesis is only the first-order approximation of the general expression.It may be unduly simplified and may fail to describe satisfactorily the damage phenomena of practical materials.

  8. Recombinant vectors construction for cellobiohydrolase encoding gene constitutive expression

    Directory of Open Access Journals (Sweden)

    Leontina GURGU

    2012-12-01

    Full Text Available Cellobiohydrolases (EC 3.2.1.91 are important exo enzymes involved in cellulose hydrolysis alongside endoglucanases (EC 3.2.1.4 and β-glucosidases (EC 3.2.1.21. Heterologous cellobiohydrolase gene expression under constitutive promoter control using Saccharomyces cerevisiae as host system is of great importance for a successful SSF process. From this point of view, the main objective of the work was to use Yeplac181 expression vector as a recipient for cellobiohdrolase - cbhB encoding gene expression under the control of the actin promoter, in Saccharomyces cerevisiae. Two hybridvectors, YEplac-Actp and YEplac-Actp-CbhB, were generated usingEscherichia coli XLI Blue for the cloning experiments. Constitutive cbhB gene expression was checked by proteine gel electrophoresis (SDS-PAGE after insertion of these constructs into Saccharomyces cerevisiae.

  9. Constitutive versus responsive gene expression strategies for growth in changing environments.

    Directory of Open Access Journals (Sweden)

    Nico Geisel

    Full Text Available Microbes respond to changing environments by adjusting gene expression levels to the demand for the corresponding proteins. Adjusting protein levels is slow, consequently cells may reach the optimal protein level only by a time when the demand changed again. It is therefore not a priori clear whether expression "on demand" is always the optimal strategy. Indeed, many genes are constitutively expressed at intermediate levels, which represents a permanent cost but provides an immediate benefit when the protein is needed. Which are the conditions that select for a responsive or a constitutive expression strategy, what determines the optimal constitutive expression level in a changing environment, and how is the fitness of the two strategies affected by gene expression noise? Based on an established model of the lac- and gal-operon expression dynamics, we study the fitness of a constitutive and a responsive expression strategy in time-varying environments. We find that the optimal constitutive expression level differs from the average demand for the gene product and from the average optimal expression level; depending on the shape of the growth rate function, the optimal expression level either provides intermediate fitness in all environments, or maximizes fitness in only one of them. We find that constitutive expression can provide higher fitness than responsive expression even when regulatory machinery comes at no cost, and we determine the minimal response rate necessary for "expression on demand" to confer a benefit. Environmental and inter-cellular noise favor the responsive strategy while reducing fitness of the constitutive one. Our results show the interplay between the demand-frequency for a gene product, the genetic response rate, and the fitness, and address important questions on the evolution of gene regulation. Some of our predictions agree with recent yeast high throughput data, for others we propose the experiments that are needed

  10. Hydra constitutively expresses transcripts involved in vertebrate neural differentiation

    Indian Academy of Sciences (India)

    Sandipan Chatterjee; Shweta Lahudkar; N N Godbole; Surendra Ghaskadbi

    2001-06-01

    The diploblastic Hydra is among the most primitive multicellular organisms. Using cross-hybridization with Xenopus probes, noggin-like transcripts were detected in the hypostome and basal disc of adult Hydra (Pelmatohydra oligactis), regions with properties similar to that of the amphibian organizer. This points to the possibility of a close molecular similarity between the Xenopus and Hydra organizers. The constitutive expression of a noggin-like gene in Hydra may be responsible for its regenerative capacity.

  11. Analytic expressions for the constitutive parameters of magnetoelectric metamaterials.

    Science.gov (United States)

    Smith, D R

    2010-03-01

    Electromagnetic metamaterials are artificially structured media typically composed of arrays of resonant electromagnetic circuits, the dimension and spacing of which are considerably smaller than the free-space wavelengths of operation. The constitutive parameters for metamaterials, which can be obtained using full-wave simulations in conjunction with numerical retrieval algorithms, exhibit artifacts related to the finite size of the metamaterial cell relative to the wavelength. Liu [R. Liu, T. J. Cui, D. Huang, B. Zhao, and D. R. Smith, Phys. Rev. E 76, 026606 (2007)] showed that the complicated, frequency-dependent forms of the constitutive parameters can be described by a set of relatively simple analytical expressions. These expressions provide useful insight and can serve as the basis for more intelligent interpolation or optimization schemes. Here, we show that the same analytical expressions can be obtained using a transfer-matrix formalism applied to a one-dimensional periodic array of thin, resonant, dielectric, or magnetic sheets. The transfer-matrix formalism breaks down, however, when both electric and magnetic responses are present in the same unit cell, as it neglects the magnetoelectric coupling between unit cells [C. R. Simovski, Metamaterials 1, 62 (2007)]. We show that an alternative analytical approach based on the same physical model must be applied for such structures. Furthermore, in addition to the intercell coupling, electric and magnetic resonators within a unit cell may also exhibit magnetoelectric coupling. For such cells, we find an analytical expression for the effective index, which displays markedly characteristic dispersion features that depend on the strength of the coupling coefficient. We illustrate the applicability of the derived expressions by comparing to full-wave simulations on magnetoelectric unit cells. We conclude that the design of metamaterials with tailored simultaneous electric and magnetic response-such as negative

  12. Living Democracy: How Constitution High School Molds Better Citizens

    Science.gov (United States)

    Brasof, Marc

    2009-01-01

    Philadelphia's Constitution High School (CHS) is committed both to the theory of education for democracy, and to its practice, as reflected by a school constitution, student elections, town hall meetings, and active student participation in school government. As its name indicates, CHS is a theme-based high school that focuses on history,…

  13. The constitutional right to freedom of expression: How enforceable ...

    African Journals Online (AJOL)

    Erna Kinsey

    realisation, one of the ... same time flouting the Constitution by denying learners' constitutional rights to dignity and ... asked in developing rules and/or deciding on whether particular beha- viour (or dress) ..... Unpublished PhD thesis. Bloemfontein: ...

  14. Tumorigenesis induced by the HHV8-encoded chemokine receptor requires ligand modulation of high constitutive activity

    DEFF Research Database (Denmark)

    Holst, P J; Rosenkilde, M M; Manfra, D;

    2001-01-01

    ORF74 (or KSHV-vGPCR) is a highly constitutively active G protein-coupled receptor encoded by HHV8 that is regulated both positively and negatively by endogenous chemokines. When expressed in transgenic mice, this chemokine receptor induces an angioproliferative disease closely resembling Kaposi...... sarcoma (KS). Here we demonstrate that several lines of mice carrying mutated receptors deficient in either constitutive activity or chemokine regulation fail to develop KS-like disease. In addition, animals expressing a receptor that preserves chemokine binding and constitutive activity but that does...

  15. Constitutive and regulated expression vectors to construct polyphosphate deficient bacteria

    Directory of Open Access Journals (Sweden)

    Jerez Carlos A

    2009-03-01

    Full Text Available Abstract Background Inorganic polyphosphate (polyP, a polymer of tens or hundreds of phosphate residues linked by ATP-like bonds, is found in all organisms and performs a wide variety of functions. PolyP is synthesized in bacterial cells by the actions of polyphosphate kinases (PPK1 and PPK2 and degraded by an exopolyphosphatase (PPX. Bacterial cells with polyP deficiencies are impaired in many structural and important cellular functions such as motility, quorum sensing, biofilm formation and virulence. Knockout mutants of the ppk1 gene have been the most frequent strategy employed to generate polyP deficient cells. Results As an alternative method to construct polyP-deficient bacteria we developed constitutive and regulated broad-host-range vectors for depleting the cellular polyP content. This was achieved by the overexpression of yeast exopolyphosphatase (PPX1. Using this approach in a polyphosphate accumulating bacteria (Pseudomonas sp. B4, we were able to eliminate most of the cellular polyP (>95%. Furthermore, the effect of overexpression of PPX1 resembled the functional defects found in motility and biofilm formation in a ppk1 mutant from Pseudomonas aeruginosa PAO1. The plasmids constructed were also successfully replicated in other bacteria such as Escherichia coli, Burkholderia and Salmonella. Conclusion To deplete polyP contents in bacteria broad-host-range expression vectors can be used as an alternative and more efficient method compared with the deletion of ppk genes. It is of great importance to understand why polyP deficiency affects vital cellular processes in bacteria. The construction reported in this work will be of great relevance to study the role of polyP in microorganisms with non-sequenced genomes or those in which orthologs to ppk genes have not been identified.

  16. Right to Freedom of Speech and Expression (constitutional legal framework and Constitutional Court practice in the Republic of Croatia

    Directory of Open Access Journals (Sweden)

    Mato Arlović

    2016-05-01

    Full Text Available This paper analyses the connection and inter relationship of freedom as the highest value of constitutional order of the Republic of Croatia and the highest ideal of contemporary democracy and constitutional state with the right to freedom of speech and expression. By analysing that interrelationship, the intention to confirm the thesis that the right to freedom of speech and expression is just one (albeit very importanthuman right and fundamental freedom arises. It has been extracted from freedoms as the highest value and as one which is always returned to in such a way that affects the scope and content of it being achieved in relation to the imagined ideal of freedom and the given reality. The right to freedom of speech and expression is analysed as just one of the fundamental supports for building a democratic society of rule of law and Constitution. In this context, the function of the indispensable condition for achieving other human rights and fundamental freedoms is also analysed. Furthermore, it is considered as a right within which, through its content, includes a range of other rights and freedoms. It should be considered from a theoretical viewpoint as a so called legal construction. This paper analyses the constitutional legal framework in the Republic of Croatia related to the right to freedom of speech and expression. In that analysis, particular attention is given to the subject, content and characteristics of the right to freedom of speech and expression and the practice of European Court of Human Rights and the Constitutional Court of the Republic of Croatia which protect this right.

  17. Evolutionary change from induced to constitutive expression of an indirect plant resistance.

    Science.gov (United States)

    Heil, Martin; Greiner, Sabine; Meimberg, Harald; Krüger, Ralf; Noyer, Jean-Louis; Heubl, Günther; Linsenmair, K Eduard; Boland, Wilhelm

    2004-07-08

    Induced plant resistance traits are expressed in response to attack and occur throughout the plant kingdom. Despite their general occurrence, the evolution of such resistances has rarely been investigated. Here we report that extrafloral nectar, a usually inducible trait, is constitutively secreted by Central American Acacia species that are obligately inhabited by ants. Extrafloral nectar is secreted as an indirect resistance, attracting ants that defend plants against herbivores. Leaf damage induces extrafloral nectar secretion in several plant species; among these are various Acacia species and other Fabaceae investigated here. In contrast, Acacia species obligately inhabited by symbiotic ants nourish these ants by secreting extrafloral nectar constitutively at high rates that are not affected by leaf damage. The phylogeny of the genus Acacia and closely related genera indicate that the inducibility of extrafloral nectar is the plesiomorphic or 'original' state, whereas the constitutive extrafloral nectar flow is derived within Acacia. A constitutive resistance trait has evolved from an inducible one, obviously in response to particular functional demands.

  18. Expression Profiling of Transcriptome and Its Associated Disease Risk in Yang Deficiency Constitution of Healthy Subjects

    Directory of Open Access Journals (Sweden)

    Ruoxi Yu

    2016-01-01

    Full Text Available Objectives. Differences among healthy subjects and associated disease risks are of substantial interest in clinical medicine. According to the theory of “constitution-disease correlation” in traditional Chinese medicine, we try to find out if there is any connection between intolerance of cold in Yang deficiency constitution and molecular evidence and if there is any gene expression basis in specific disorders. Methods. Peripheral blood mononuclear cells were collected from Chinese Han individuals with Yang deficiency constitution (n=20 and balanced constitution (n=8 (aged 18–28 and global gene expression profiles were determined between them using the Affymetrix HG-U133 Plus 2.0 array. Results. The results showed that when the fold change was ≥1.2 and q ≤ 0.05, 909 genes were upregulated in the Yang deficiency constitution, while 1189 genes were downregulated. According to our research differential genes found in Yang deficiency constitution were usually related to lower immunity, metabolic disorders, and cancer tendency. Conclusion. Gene expression disturbance exists in Yang deficiency constitution, which corresponds to the concept of constitution and gene classification. It also suggests people with Yang deficiency constitution are susceptible to autoimmune diseases, enteritis, arthritis, metabolism disorders, and cancer, which provides molecular evidence for the theory of “constitution-disease correlation.”

  19. Efficient production and evaluation of lignocellulolytic enzymes using a constitutive protein expression system in Penicillium oxalicum.

    Science.gov (United States)

    Hu, Yibo; Xue, Haizhao; Liu, Guodong; Song, Xin; Qu, Yinbo

    2015-06-01

    Native lignocellulolytic enzyme systems secreted by filamentous fungi can be further optimized by protein engineering or supplementation of exogenous enzyme components. We developed a protein production and evaluation system in cellulase-producing fungus Penicillium oxalicum. First, by deleting the major amylase gene amy15A, a strain Δ15A producing few extracellular proteins on starch was constructed. Then, three lignocellulolytic enzymes (BGL4, Xyn10B, and Cel12A) with originally low expression levels were successfully expressed with selected constitutive promoters in strain Δ15A. BGL4 and Cel12A overexpression resulted in increased specific filter paper activity (FPA), while the overexpression of Xyn10B improved volumetric FPA but not specific FPA. By switching the culture medium, this platform is convenient to produce originally low-expressed lignocellulolytic enzymes in relatively high purities on starch and to evaluate the effect of their supplementation on the performance of a complex cellulase system on cellulose.

  20. Cloning and constitutive expression of Deschampsia antarctica Cu/Zn superoxide dismutase in Pichia pastoris

    Directory of Open Access Journals (Sweden)

    Gidekel Manuel

    2009-10-01

    Full Text Available Abstract Background Deschampsia antarctica shows tolerance to extreme environmental factors such as low temperature, high light intensity and an increasing UV radiation as result of the Antarctic ozone layer thinning. It is very likely that the survival of this species is due to the expression of genes that enable it to tolerate high levels of oxidative stress. On that account, we planned to clone the D. antarctica Cu/ZnSOD gene into Pichia pastoris and to characterize the heterologous protein. Findings The Copper/Zinc superoxide dismutase (Cu/ZnSOD gene, SOD gene, was isolated from a D. antarctica by cDNA library screening. This SOD gene was cloned in the expression vector pGAPZαA and successfully integrated into the genome of the yeast P. pastoris SMD1168H. A constitutive expression system for the expression of the recombinant SOD protein was used. The recombinant protein was secreted into the YPD culture medium as a glycosylated protein with a 32 mg/l expression yield. The purified recombinant protein possesses a specific activity of 440 U/mg. Conclusion D. antarctica Cu/ZnSOD recombinant protein was expressed in a constitutive system, and purified in a single step by means of an affinity column. The recombinant SOD was secreted to the culture medium as a glycoprotein, corresponding to approximately 13% of the total secreted protein. The recombinant protein Cu/ZnSOD maintains 60% of its activity after incubation at 40°C for 30 minutes and it is stable (80% of activity between -20°C and 20°C. The recombinant SOD described in this study can be used in various biotechnological applications.

  1. Constitutive expression of human pancreatic lipase-related protein 1 in Pichia pastoris.

    Science.gov (United States)

    Aloulou, Ahmed; Grandval, Philippe; De Caro, Josiane; De Caro, Alain; Carrière, Frédéric

    2006-06-01

    High-level constitutive expression of the human pancreatic lipase-related protein 1 (HPLRP1) was achieved using the methylotrophic yeast Pichia pastoris. The HPLRP1 cDNA, including its original leader sequence, was subcloned into the pGAPZB vector and further integrated into the genome of P. pastoris X-33 under the control of the glyceraldehyde 3-phosphate dehydrogenase (GAP) constitutive promoter. A major protein with a molecular mass of 50 kDa was found to be secreted into the culture medium and was identified using anti-HPLRP1 polyclonal antibodies as HPLRP1 recombinant protein. The level of expression reached 100-120 mg of HPLRP1 per liter of culture medium after 40 h, as attested by specific and quantitative enzyme-linked immunosorbent assay. A single cation-exchange chromatography sufficed to obtain a highly purified recombinant HPLRP1 after direct batch adsorption onto S-Sepharose of the HPLRP1 present in the culture medium, at pH 5.5. N-terminal sequencing and mass spectrometry analysis were carried out to monitor the production of the mature protein and to confirm that its signal peptide was properly processed.

  2. A recursive network approach can identify constitutive regulatory circuits in gene expression data

    Science.gov (United States)

    Blasi, Monica Francesca; Casorelli, Ida; Colosimo, Alfredo; Blasi, Francesco Simone; Bignami, Margherita; Giuliani, Alessandro

    2005-03-01

    The activity of the cell is often coordinated by the organisation of proteins into regulatory circuits that share a common function. Genome-wide expression profiles might contain important information on these circuits. Current approaches for the analysis of gene expression data include clustering the individual expression measurements and relating them to biological functions as well as modelling and simulation of gene regulation processes by additional computer tools. The identification of the regulative programmes from microarray experiments is limited, however, by the intrinsic difficulty of linear methods to detect low-variance signals and by the sensitivity of the different approaches. Here we face the problem of recognising invariant patterns of correlations among gene expression reminiscent of regulation circuits. We demonstrate that a recursive neural network approach can identify genetic regulation circuits from expression data for ribosomal and genome stability genes. The proposed method, by greatly enhancing the sensitivity of microarray studies, allows the identification of important aspects of genetic regulation networks and might be useful for the discrimination of the different players involved in regulation circuits. Our results suggest that the constitutive regulatory networks involved in the generic organisation of the cell display a high degree of clustering depending on a modular architecture.

  3. Brief Considerations on the German Constitutional Approach of the Freedom of Expression

    Directory of Open Access Journals (Sweden)

    Carmen MOLDOVAN

    2015-08-01

    Full Text Available The process of understanding the concept of freedom of expression is very complex and it implies thorough knowledge of the constitutional values and the analysis of the constitutional tradition of different legal systems. At the same time, a balancing operation is necessary between the value of freedom of speech and other fundamental rights, process that establishes limits of the expression such as criminalizing hate speech or other actions which affect the individual dignity. The human dignity has become a constant component of the constitutional case law of many States that share the same characteristics due to the promotion of fundamental rights. In the absence of a hierarchy of rights and fundamental freedom, freedom of expression is analyzed in many cases as a primary right (Kentridge, 1996, p. 254 that prevails over other fundamental rights. This study aims to analyze the landmarks of the German constitutional interpretation of the concept of freedom of expression, to identify its scope and types of speech excluded from the constitutional protection. The choice of this constitutional legal order was justified by the complexity of this topic and the specificity of the German history and tradition, elements that have contributed to an interesting vision in the matter of conflict between different fundamental rights.

  4. Constitutive expression of gamma-H2AX has prognostic relevance in triple negative breast cancer

    NARCIS (Netherlands)

    Nagelkerke, A.P.; Kuijk, S.J. van; Sweep, F.C.; Nagtegaal, I.D.; Hoogerbrugge, N.; Martens, J.W.; Timmermans, M.A.Y.; Laarhoven, H.W.M. van; Bussink, J.; Span, P.N.

    2011-01-01

    BACKGROUND AND PURPOSE: Constitutive gamma-H2AX expression might indicate disruption of the DNA damage repair pathway, genomic instability, or shortened telomeric ends. Here, we quantified expression of endogenous gamma-H2AX and its downstream factor 53BP1 in a large number of breast cancer cell lin

  5. STAT3-mediated constitutive expression of SOCS-3 in cutaneous T-cell lymphoma

    DEFF Research Database (Denmark)

    Brender, C; Nielsen, M; Kaltoft, K;

    2001-01-01

    A characteristic feature of neoplastic transformation is the loss of external control by cytokines and extracellular matrix of cellular differentiation, migration, and mitogenesis. Because suppressors of cytokine signaling (SOCS) proteins are negative regulators of cytokine-induced signaling......, it has been hypothesized that an aberrant SOCS expression plays a role in neoplastic transformation. This study reports on a constitutive SOCS-3 expression in cutaneous T-cell lymphoma (CTCL) cell lines. SOCS-3 protein is constitutively expressed in tumor cell lines (but not in nonmalignant T cells......) obtained from affected skin from a patient with mycosis fungoides (MF) and from peripheral blood from a patient with Sezary syndrome (SS). In contrast, constitutive SOCS-3 expression is not found in the leukemic Jurkat T-cell line, the MOLT-4 acute lymphoblastic leukemia cell line, and the monocytic...

  6. Constitutive SOCS-3 expression protects T-cell lymphoma against growth inhibition by IFNalpha

    DEFF Research Database (Denmark)

    Brender, C; Lovato, P; Sommer, V H;

    2005-01-01

    Signal transducer and activator of transcription (Stat)3 is constitutively activated in cutaneous T-cell lymphoma (CTCL), where it protects tumour cells against apoptosis. The constitutive activation of Stat3 leads to a constitutive expression of suppressor of cytokine signalling (SOCS)-3....... In healthy cells, SOCS-3 is transiently expressed following cytokine stimulation and functions as a negative feedback inhibitor of the Stat3-activating kinases. Here, we attempt to resolve the apparent paradox of a simultaneous SOCS-3 expression and Stat3 activation in the same cells. We show that (i) SOCS-3...... expression in tumour cells is equal to or higher than in cytokine-stimulated nonmalignant T cells, (ii) SOCS-3 is not mutated in CTCL, (iii) overexpression of SOCS-3 blocks IFNalpha-mediated growth inhibition without affecting Stat3 activation, growth, and apoptosis, and (iv) inhibition of SOCS-3...

  7. Identification and refinement of two strong constitutive promoters for gene expression system of Schizosaccharomyces pombe.

    Science.gov (United States)

    Wang, Hongcheng; Wang, Haiyang; Wang, Meng; Zhang, Lei; Wang, Ren; Mei, Yanzhen; Shao, Weilan

    2014-06-01

    Fission yeast Schizosaccharomyces pombe shares various important properties with higher eukaryotes and is now considered a useful host for elevated production of mammalian proteins for medicinal applications. The full-length nmt1 promoter has been widely used as a strong promoter in S. pombe expression system. In the present study, the promoters of the eno101 and gpd3 genes in S. pombe were identified as strong constitutive promoters. For convenient applications in the plasmids of S. pombe, these promoters were refined to 276-bp eno and 273-bp gpd promoters by deleting undesired sequences and examining the expression of reporter genes including lacZ and xynA. Both the refined eno and gpd promoters provided approximately 1.5-fold higher expression of LacZ than nmt1 promoter. Furthermore, gene expression under the control of the eno or gpd promoter was not repressed by the components of YES medium while nmt1 promoter was inhibited by thiamine in yeast extract. Therefore, both eno and gpd promoters offer opportunities for efficient production of recombinant proteins by S. pombe in high cell-density fermentation.

  8. Constitutive expression and activity of cytochrome P450 in conventional pigs.

    Science.gov (United States)

    Nielsen, Søren Drud; Bauhaus, Yvonne; Zamaratskaia, Galia; Junqueira, Matheus Antunes; Blaabjerg, Karoline; Petrat-Melin, Bjørn; Young, Jette Feveile; Rasmussen, Martin Krøyer

    2017-04-01

    Pigs have often been suggested to be a useful model for humans, when investigating CYP dependent events, like drug metabolism. However, comprehensive knowledge about the constitutive expression of the major CYP and corresponding transcription factors is limited. We compared the constitutive mRNA expression of aryl hydrocarbon receptor, constitutive androstane receptor and pregnane X receptor and CYP1A1, CYP1A2, CYP2A, CYP2E1 and CYP3A in liver, adipose tissue, muscle and small intestine in pigs, as well as the expression along the length of the small intestine and colon. Tissue samples were taken from female pigs, and analyzed for gene expression, as well as CYP dependent activity using qPCR and specific probe substrates, respectively. For all investigated transcription factors and CYPs the mRNA expression and activity was highest in the liver. CYP1A1 and CYP3A mRNA expression and activity was shown in all investigated tissues. Along the small intestine and colon the mRNA expression and activity of CYP1A1 and CYP3A was gradually decreased. The results demonstrated, similarity to that reported for humans, and hence adds to the use of pigs as a model for humans.

  9. A homolog of the RPS2 disease resistance gene is constitutively expressed in Brassica oleracea

    Directory of Open Access Journals (Sweden)

    Malvas Celia C.

    2003-01-01

    Full Text Available In this study, we identified disease resistance gene homologs in Brassica oleracea and assessed their expression in lines resistant and susceptible to Xanthomonas campestris pv. campestris (Xcc. Two DNA fragments of approximately 2.5 kb (BI-16/RPS2 and Lc201/RPS2 were amplified by PCR from two Brassica lines using primers based on an RPS2 homologous sequence previously described in the Brassica oleracea ecotype B117. The sequences of these fragments shared high similarity (95-98% with RPS2 homologs from various Brassica species. The digestion of these fragments with restriction enzymes revealed polymorphisms at the Xba I restriction sites. The length polymorphisms were used as a co-dominant marker in an F2 population developed to segregate for resistance to Xcc, the causal agent of black rot. Linkage analysis showed no significant association between the marker and quantitative trait loci for black rot. RT-PCR with specific primers yielded an expected 453 bp fragment that corresponded to the RPS2 homologs in both resistant and susceptible lines inoculated with the pathogen, as well as in non-inoculated control plants. These results suggest that these homologs are constitutively expressed in B. oleracea.

  10. Carbon Costs of Constitutive and Expressed Resistance to a Non-Native Pathogen in Limber Pine

    Science.gov (United States)

    2016-01-01

    Increasing the frequency of resistance to the non-native fungus Cronartium ribicola (causative agent of white pine blister rust, WPBR) in limber pine populations is a primary management objective to sustain high-elevation forest communities. However, it is not known to what extent genetic disease resistance is costly to plant growth or carbon economy. In this study, we measured growth and leaf-level physiology in (1) seedling families from seed trees that have previously been inferred to carry or not carry Cr4, the dominant R gene allele conferring complete, gene-for-gene resistance to WPBR in limber pine, and (2) populations that were and were not infected with C. ribicola. We found that, in the absence of C. ribicola exposure, there was no significant difference in carbon relations between families born from seed trees that harbor the resistance allele compared to those that lack it, either to plant growth and phenology or leaf-level photosynthetic traits. However, post-infection with C. ribicola, growth was significantly reduced in inoculation survivors expressing complete resistance compared to uninoculated seedlings. Furthermore, inoculation survivors exhibited significant increases in a suite of traits including photosynthetic rate, respiration rate, leaf N, and stomatal conductance and a decrease in photosynthetic water-use efficiency. The lack of constitutive carbon costs associated with Cr4 resistance in non-stressed limber pine is consistent with a previous report that the R gene allele is not under selection in the absence of C. ribicola and suggests that host resistance may not bear a constitutive cost in pathosystems that have not coevolved. However, under challenge by C. ribicola, complete resistance to WPBR in limber pine has a significant cost to plant growth, though enhanced carbon acquisition post-infection may offset this somewhat. These costs and effects on performance further complicate predictions of this species’ response in warmer future

  11. Constitutive expression of Yarrowia lipolytica lipase LIP2 in Pichia pastoris using GAP as promoter.

    Science.gov (United States)

    Wang, Xiaofeng; Sun, Yongchuan; Ke, Feng; Zhao, Heyun; Liu, Tao; Xu, Li; Liu, Yun; Yan, Yunjun

    2012-03-01

    A gene encoding Yarrowia lipolytica lipase LIP2 (YlLIP2) was cloned into a constitutive expression vector pGAPZαA and electrotransformed into the Pichia pastoris X-33 strain. The high-yield clones obtained by high copy and enzyme activity screening were chosen as the host strains for shaking flask and fermentor culture. The results showed that glucose was the optimum carbon source for YlLIP2 production, and the maximum hydrolytic activity of recombinant YlLIP2 reached 1,315 U/ml under the flask culture at 28 °C, pH 7.0, for 48 h. The fed-batch fermentation was carried out in 3- and 10-l bioreactors by continuously feeding glucose into the growing medium for achieving high cell density and YlLIP2 yields. The maximum hydrolytic activity of YlLIP2 and cell density obtained in the 3-l bioreactor were 10,300 U/ml and 116 g dry cell weight (DCW)/l, respectively. The peak hydrolytic activity of YlLIP2 and cell density were further improved in the 10-l fermentor where the values respectively attained were 13,500 U/ml and 120 g DCW/l. The total protein concentration in the supernatant reached 3.3 g/l and the cell viability remained approximately 99% after 80 h of culture. Furthermore, the recombinant YlLIP2 produced in P. pastoris pGAP and pAOX1 systems have similar content of sugar (about 12%) and biochemical characteristics. The above results suggest that the GAP promoter-derived expression system of P. pastoris is effective for the expression of YlLIP2 by high cell density culture and is probably an alternative to the conventional AOX1 promoter expression system in large-scale production of industrial lipases.

  12. Selective elimination of high constitutive activity or chemokine binding in the human herpesvirus 8 encoded seven transmembrane oncogene ORF74

    DEFF Research Database (Denmark)

    Rosenkilde, M M; Kledal, T N; Holst, Peter Johannes;

    2000-01-01

    expressing ORF74 under control of the CD2 promoter develop highly vascularized Kaposi's sarcoma-like tumors. Through targeted mutagenesis we here create three distinct phenotypes of ORF74: a receptor with normal, high constitutive signaling through the phospholipase C pathway but deprived of binding...

  13. Gene expression of two kinds of constitutive nitric oxide synthase in injured spinal cord tissue

    Institute of Scientific and Technical Information of China (English)

    刘成龙; 靳安民; 周初松; 闵少雄

    2002-01-01

    Objective: To investigate the gene expression of two kinds of constitutive nitric oxide synthase (cNOS): neuronal NOS (nNOS) and endothelial NOS (eNOS) in injured spinal cord tissue.   Methods: Thirty-six adult Sprague-Dawley rats were divided randomly into six groups: the normal group and the injury groups (2, 6, 12, 24, 48 h after injury, respectively). A compression injury model of the spinal cord was made and gene expression of nNOS and eNOS were examined by reverse transcription polymerase chain reaction (RT-PCR).   Results: The gene expression of nNOS and eNOS was detected in the normal group and they were up-regulated quickly after injury, reaching the maximum at 6 h. There was no difference between gene expression of nNOS and eNOS in the normal group, but in each injury group the gene expression of eNOS was much higher than that of nNOS.   Conclusions: Expression of constitutive NOS (cNOS) in spinal cord tissue was up-regulated after injury mainly in the early stage. cNOS as a whole offers protection in spinal cord injury, but different cNOS may play different roles.

  14. Constitutive and Inducible Expression of Green Fluorescent Protein in Brucella suis

    Science.gov (United States)

    Köhler, Stephan; Ouahrani-Bettache, Safia; Layssac, Marion; Teyssier, Jacques; Liautard, Jean-Pierre

    1999-01-01

    A gene fusion system based on plasmid pBBR1MCS and the expression of green fluorescent protein was developed for Brucella suis, allowing isolation of constitutive and inducible genes. Bacteria containing promoter fusions of chromosomal DNA to gfp were visualized by fluorescence microscopy and examined by flow cytometry. Twelve clones containing gene fragments induced inside J774 murine macrophages were isolated and further characterized. PMID:10569794

  15. Constitutive and Inducible Expression of Green Fluorescent Protein in Brucella suis

    OpenAIRE

    Köhler, Stephan; Ouahrani-Bettache, Safia; Layssac, Marion; Teyssier, Jacques; Liautard, Jean-Pierre

    1999-01-01

    A gene fusion system based on plasmid pBBR1MCS and the expression of green fluorescent protein was developed for Brucella suis, allowing isolation of constitutive and inducible genes. Bacteria containing promoter fusions of chromosomal DNA to gfp were visualized by fluorescence microscopy and examined by flow cytometry. Twelve clones containing gene fragments induced inside J774 murine macrophages were isolated and further characterized.

  16. Identification of chemical modulators of the constitutive activated receptor (CAR) in a gene expression compendium

    OpenAIRE

    Oshida, Keiyu; Vasani, Naresh; Jones, Carlton; Moore, Tanya; Hester, Susan; Nesnow, Stephen; Auerbach, Scott; Geter, David R.; Aleksunes, Lauren M; Thomas, Russell S.; Applegate, Dawn; Klaassen, Curtis D.; Corton, J. Christopher

    2015-01-01

    The nuclear receptor family member constitutive activated receptor (CAR) is activated by structurally diverse drugs and environmentally-relevant chemicals leading to transcriptional regulation of genes involved in xenobiotic metabolism and transport. Chronic activation of CAR increases liver cancer incidence in rodents, whereas suppression of CAR can lead to steatosis and insulin insensitivity. Here, analytical methods were developed to screen for chemical treatments in a gene expression comp...

  17. Constitutively Expressed αB—Crystallin in Heat Schock Transcription Factor 1 Knockout Mice Myocardium

    Institute of Scientific and Technical Information of China (English)

    刘莉; 张洪慧; 丁国宪; 程蕴琳; 晏良军; BENJAMINIvorJ

    2003-01-01

    Objective:To invesligate the effects of heat shock transcription factor 1(HSF 1) gene on the constitutively expressed aB-Crystallin(αBC)in mice myocardium.Methods:The expression levels of constitutive αBC in HSF1 knockout(hsf1-/-) and HSF1 wild type (hsf1+/+) mice myocardium were evaluated by western blot and immunohistochemistry.Results:The αBC levels in hsf1-/- and hsf1+/_ were 68.42±4.16,100.00±7.58,respectively(P<0.05,cytosolic fraction),and 20.53±1.01,37.55±1.91,respectively(P<0.05,pellet fraction).The αBC signals decreased significantly in hsf1-/- myocardium when compared with those in hsf1+/+ myocardium stained with fluorescence immunohistochemistry.Conclusion.HSF1 is an important,but not the only factor,which mediates the constitutively expressed αBC.

  18. Constitutive type VI secretion system expression gives Vibrio cholerae intra- and interspecific competitive advantages.

    Directory of Open Access Journals (Sweden)

    Daniel Unterweger

    Full Text Available The type VI secretion system (T6SS mediates protein translocation across the cell membrane of Gram-negative bacteria, including Vibrio cholerae - the causative agent of cholera. All V. cholerae strains examined to date harbor gene clusters encoding a T6SS. Structural similarity and sequence homology between components of the T6SS and the T4 bacteriophage cell-puncturing device suggest that the T6SS functions as a contractile molecular syringe to inject effector molecules into prokaryotic and eukaryotic target cells. Regulation of the T6SS is critical. A subset of V. cholerae strains, including the clinical O37 serogroup strain V52, express T6SS constitutively. In contrast, pandemic strains impose tight control that can be genetically disrupted: mutations in the quorum sensing gene luxO and the newly described regulator gene tsrA lead to constitutive T6SS expression in the El Tor strain C6706. In this report, we examined environmental V. cholerae isolates from the Rio Grande with regard to T6SS regulation. Rough V. cholerae lacking O-antigen carried a nonsense mutation in the gene encoding the global T6SS regulator VasH and did not display virulent behavior towards Escherichia coli and other environmental bacteria. In contrast, smooth V. cholerae strains engaged constitutively in type VI-mediated secretion and displayed virulence towards prokaryotes (E. coli and other environmental bacteria and a eukaryote (the social amoeba Dictyostelium discoideum. Furthermore, smooth V. cholerae strains were able to outcompete each other in a T6SS-dependent manner. The work presented here suggests that constitutive T6SS expression provides V. cholerae with an advantage in intraspecific and interspecific competition.

  19. Constitutive type VI secretion system expression gives Vibrio cholerae intra- and interspecific competitive advantages.

    Science.gov (United States)

    Unterweger, Daniel; Kitaoka, Maya; Miyata, Sarah T; Bachmann, Verena; Brooks, Teresa M; Moloney, Jessica; Sosa, Oscar; Silva, David; Duran-Gonzalez, Jorge; Provenzano, Daniele; Pukatzki, Stefan

    2012-01-01

    The type VI secretion system (T6SS) mediates protein translocation across the cell membrane of Gram-negative bacteria, including Vibrio cholerae - the causative agent of cholera. All V. cholerae strains examined to date harbor gene clusters encoding a T6SS. Structural similarity and sequence homology between components of the T6SS and the T4 bacteriophage cell-puncturing device suggest that the T6SS functions as a contractile molecular syringe to inject effector molecules into prokaryotic and eukaryotic target cells. Regulation of the T6SS is critical. A subset of V. cholerae strains, including the clinical O37 serogroup strain V52, express T6SS constitutively. In contrast, pandemic strains impose tight control that can be genetically disrupted: mutations in the quorum sensing gene luxO and the newly described regulator gene tsrA lead to constitutive T6SS expression in the El Tor strain C6706. In this report, we examined environmental V. cholerae isolates from the Rio Grande with regard to T6SS regulation. Rough V. cholerae lacking O-antigen carried a nonsense mutation in the gene encoding the global T6SS regulator VasH and did not display virulent behavior towards Escherichia coli and other environmental bacteria. In contrast, smooth V. cholerae strains engaged constitutively in type VI-mediated secretion and displayed virulence towards prokaryotes (E. coli and other environmental bacteria) and a eukaryote (the social amoeba Dictyostelium discoideum). Furthermore, smooth V. cholerae strains were able to outcompete each other in a T6SS-dependent manner. The work presented here suggests that constitutive T6SS expression provides V. cholerae with an advantage in intraspecific and interspecific competition.

  20. Gonadotropin-releasing hormone is prerequisite for the constitutive expression of pituitary annexin A5.

    Science.gov (United States)

    Yonezawa, Tomohiro; Watanabe, Aiko; Kurusu, Shiro; Kawaminami, Mitsumori

    2015-01-01

    Annexin A5 (ANXA5), a member of the structurally related family of annexin proteins, is expressed in pituitary gonadotropes. We previously reported that ANXA5 expression is stimulated by gonadotropin-releasing hormone (GnRH). In the present study, we investigated ANXA5 expression in the anterior pituitary gland of GnRH-deficient mutant hypogonadal (hpg) mice. RT-PCR demonstrated that luteinizing hormone β subunit (LHβ) and ANXA5 mRNA levels were both lower in the pituitary gland of hpg mice than in wild-type mice. Immunohistochemistry showed that ANXA5 expression throughout the pituitary gland was very low in hpg mice, suggesting that ANXA5 is diminished in gonadotropes and also in other cell types. Subcutaneous administration of a GnRH analogue, des-gly10 (Pro9)-GnRH ethylamide (1 μg/day for 7 days), augmented the expression of LHβ and ANXA5 in the pituitary gland in hpg mice. However, LHβ- and ANXA5-positive cells did not show exactly matched spatial distributions. These findings suggest that GnRH is necessary for constitutive ANXA5 expression in the pituitary gland, not only in gonadotropes but also in other pituitary gland cell types. A close relationship between ANXA5 and LHβ expression was confirmed. It is suggested that a significant role of ANXA5 in the physiologic secretion of LH.

  1. Human CD4+ CD25+ Foxp3+ regulatory T cells do not constitutively express IL-35.

    Science.gov (United States)

    Bardel, Emilie; Larousserie, Frédérique; Charlot-Rabiega, Pascaline; Coulomb-L'Herminé, Aurore; Devergne, Odile

    2008-11-15

    EBV-induced gene 3 (EBI3) can associate with p28 to form the heterodimeric cytokine IL-27, or with the p35 subunit of IL-12 to form the EBI3/p35 heterodimer, recently named IL-35. In mice, IL-35 has been shown to be constitutively expressed by CD4(+)CD25(+)Foxp3(+) regulatory T cells (Treg cells) and suggested to contribute to their suppressive activity. In this study, we investigated whether human Treg cells express IL-35. Double-staining analysis of human thymuses showed that neither Foxp3(+) nor CD25(+) cells coexpressed EBI3. Similarly, Foxp3(+) cells present in human lymph nodes, tonsils, spleens, and intestines did not express EBI3. Consistent with these in situ observations, Treg cells purified from blood or tonsils were negative for EBI3 by immunoblotting. Other human T cell subsets, including effector T cells, naive and memory CD4(+) T cells, CD8(+) and gammadelta T cells also did not constitutively express EBI3, which contrasts with IL-35 expression observed in murine CD8(+) and gammadelta T cells. Furthermore, although CD3/CD28 stimulation consistently induced low levels of EBI3 in various CD4(+) T cell subsets, no EBI3 could be detected in CD3/CD28-stimulated Treg cells. RT-PCR analysis showed that, whereas p35 transcripts were detected in both Teff and Treg cells, EBI3 transcripts were detected only in activated Teff cells, but not in resting or activated Treg cells. Thus, in contrast to their murine counterpart, human Treg cells do not express detectable amounts of IL-35.

  2. Conditional expression of constitutively active estrogen receptor {alpha} in chondrocytes impairs longitudinal bone growth in mice

    Energy Technology Data Exchange (ETDEWEB)

    Ikeda, Kazuhiro [Division of Gene Regulation and Signal Transduction, Research Center for Genomic Medicine, Saitama Medical University, Saitama (Japan); Tsukui, Tohru [Experimental Animal Laboratory, Research Center for Genomic Medicine, Saitama Medical University, Saitama (Japan); Imazawa, Yukiko; Horie-Inoue, Kuniko [Division of Gene Regulation and Signal Transduction, Research Center for Genomic Medicine, Saitama Medical University, Saitama (Japan); Inoue, Satoshi, E-mail: INOUE-GER@h.u-tokyo.ac.jp [Division of Gene Regulation and Signal Transduction, Research Center for Genomic Medicine, Saitama Medical University, Saitama (Japan); Department of Geriatric Medicine, Graduate School of Medicine, The University of Tokyo, Tokyo (Japan); Department of Anti-Aging Medicine, Graduate School of Medicine, The University of Tokyo, Tokyo (Japan)

    2012-09-07

    Highlights: Black-Right-Pointing-Pointer Conditional transgenic mice expressing constitutively active estrogen receptor {alpha} (caER{alpha}) in chondrocytes were developed. Black-Right-Pointing-Pointer Expression of caER{alpha} in chondrocytes impaired longitudinal bone growth in mice. Black-Right-Pointing-Pointer caER{alpha} affects chondrocyte proliferation and differentiation. Black-Right-Pointing-Pointer This mouse model is useful for understanding the physiological role of ER{alpha}in vivo. -- Abstract: Estrogen plays important roles in the regulation of chondrocyte proliferation and differentiation, which are essential steps for longitudinal bone growth; however, the mechanisms of estrogen action on chondrocytes have not been fully elucidated. In the present study, we generated conditional transgenic mice, designated as caER{alpha}{sup ColII}, expressing constitutively active mutant estrogen receptor (ER) {alpha} in chondrocytes, using the chondrocyte-specific type II collagen promoter-driven Cre transgenic mice. caER{alpha}{sup ColII} mice showed retardation in longitudinal growth, with short bone lengths. BrdU labeling showed reduced proliferation of hypertrophic chondrocytes in the proliferating layer of the growth plate of tibia in caER{alpha}{sup ColII} mice. In situ hybridization analysis of type X collagen revealed that the maturation of hypertrophic chondrocytes was impaired in caER{alpha}{sup ColII} mice. These results suggest that ER{alpha} is a critical regulator of chondrocyte proliferation and maturation during skeletal development, mediating longitudinal bone growth in vivo.

  3. A bioluminescence reporter mouse that monitors expression of constitutively active β-catenin

    Science.gov (United States)

    Kommagani, Ramakrishna; Peavey, Mary C.; Hai, Lan; Lonard, David M.; Lydon, John P.

    2017-01-01

    This short technical report describes the generation and characterization of a bioluminescence reporter mouse that is engineered to detect and longitudinally monitor the expression of doxycycline-induced constitutively active β-catenin. The new responder transgenic mouse contains the TetO-ΔN89β-CatTMILA transgene, which consists of the tet-operator followed by a bicistronic sequence encoding a stabilized form of active β-catenin (ΔN89β-catenin), an internal ribosome entry site, and the firefly luciferase gene. To confirm that the transgene operates as designed, TetO-ΔN89β-CatTMILA transgenic mouse lines were crossed with an effector mouse that harbors the mouse mammary tumor virus-reverse tetracycline transactivator (MMTV-rtTA) transgene (termed MTB hereon), which primarily targets rtTA expression to the mammary epithelium. Following doxycycline administration, the resultant MTB/CatTMILA bigenic reporter exhibited precocious lobuloalveologenesis, ductal hyperplasia, and mammary adenocarcinomas, which were visualized and monitored by in vivo bioluminescence detection. Therefore, we predict that the TetO-ΔN89β-CatTMILA transgenic responder mouse—when crossed with the appropriate effector transgenic—will have wide-applicability to non-invasively monitor the influence of constitutively active β-catenin expression on cell-fate specification, proliferation, differentiation, and neoplastic transformation in a broad spectrum of target tissues. PMID:28253313

  4. Transgenic mice expressing constitutive active MAPKAPK5 display gender-dependent differences in exploration and activity

    Directory of Open Access Journals (Sweden)

    Moens Ugo

    2007-11-01

    Full Text Available Abstract Background The mitogen-activated protein kinases, MAPKs for short, constitute cascades of signalling pathways involved in the regulation of several cellular processes that include cell proliferation, differentiation and motility. They also intervene in neurological processes like fear conditioning and memory. Since little remains known about the MAPK-Activated Protein Kinase, MAPKAPK5, we constructed the first MAPKAPK knockin mouse model, using a constitutive active variant of MAPKAPK5 and analyzed the resulting mice for changes in anxiety-related behaviour. Methods We performed primary SHIRPA observations during background breeding into the C57BL/6 background and assessed the behaviour of the background-bred animals on the elevated plus maze and in the light-dark test. Our results were analyzed using Chi-square tests and homo- and heteroscedatic T-tests. Results Female transgenic mice displayed increased amounts of head dips and open arm time on the maze, compared to littermate controls. In addition, they also explored further into the open arm on the elevated plus maze and were less active in the closed arm compared to littermate controls. Male transgenic mice displayed no differences in anxiety, but their locomotor activity increased compared to non-transgenic littermates. Conclusion Our results revealed anxiety-related traits and locomotor differences between transgenic mice expressing constitutive active MAPKAPK5 and control littermates.

  5. Contraction of the type I IFN locus and unusual constitutive expression of IFN-α in bats.

    Science.gov (United States)

    Zhou, Peng; Tachedjian, Mary; Wynne, James W; Boyd, Victoria; Cui, Jie; Smith, Ina; Cowled, Christopher; Ng, Justin H J; Mok, Lawrence; Michalski, Wojtek P; Mendenhall, Ian H; Tachedjian, Gilda; Wang, Lin-Fa; Baker, Michelle L

    2016-03-01

    Bats harbor many emerging and reemerging viruses, several of which are highly pathogenic in other mammals but cause no clinical signs of disease in bats. To determine the role of interferons (IFNs) in the ability of bats to coexist with viruses, we sequenced the type I IFN locus of the Australian black flying fox, Pteropus alecto, providing what is, to our knowledge, the first gene map of the IFN region of any bat species. Our results reveal a highly contracted type I IFN family consisting of only 10 IFNs, including three functional IFN-α loci. Furthermore, the three IFN-α genes are constitutively expressed in unstimulated bat tissues and cells and their expression is unaffected by viral infection. Constitutively expressed IFN-α results in the induction of a subset of IFN-stimulated genes associated with antiviral activity and resistance to DNA damage, providing evidence for a unique IFN system that may be linked to the ability of bats to coexist with viruses.

  6. Rho GTPase signaling promotes constitutive expression and release of TGF-β2 by human trabecular meshwork cells.

    Science.gov (United States)

    Pervan, Cynthia L; Lautz, Jonathan D; Blitzer, Andrea L; Langert, Kelly A; Stubbs, Evan B

    2016-05-01

    Elevated intraocular pressure (IOP) is causally implicated in the pathophysiology of primary open-angle glaucoma (POAG). The molecular mechanisms responsible for elevated IOP remain elusive, but may involve aberrant expression and signaling of transforming growth factor (TGF)-β2 within the trabecular meshwork (TM). Consistent with previously published studies, we show here that exogenous addition of TGF-β2 to cultured porcine anterior segments significantly attenuates outflow facility in a time-dependent manner. By comparison, perfusing segments with a TGFβRI/ALK-5 antagonist (SB-431542) unexpectedly elicited a significant and sustained increase in outflow facility, implicating a role for TM-localized constitutive expression and release of TGF-β2. Consistent with this thesis, cultured primary or transformed (GTM3) quiescent human TM cells were found to constitutively express and secrete measurable amounts of biologically-active TGF-β2. Disrupting monomeric GTPase post-translational prenylation and activation with lovastatin or GGTI-298 markedly reduced constitutive TGF-β2 expression and release. Specifically, inhibiting the Rho subfamily of GTPases with C3 exoenzyme similarly reduced constitutive expression and secretion of TGF-β2. These findings suggest that Rho GTPase signaling, in part, regulates constitutive expression and release of biologically-active TGF-β2 from human TM cells. Localized constitutive expression and release of TGF-β2 by TM cells may promote or exacerbate elevation of IOP in POAG.

  7. Constitutively expressed DHAR and MDHAR influence fruit, but not foliar ascorbate levels in tomato.

    Science.gov (United States)

    Haroldsen, Victor M; Chi-Ham, Cecilia L; Kulkarni, Shashank; Lorence, Argelia; Bennett, Alan B

    2011-10-01

    Vitamin C (L-ascorbate, AsA) is an essential nutrient required in key metabolic functions in humans and must be obtained from the diet, mainly from fruits and vegetables. Given its importance in human health and plant physiology we sought to examine the role of the ascorbate recycling enzymes monodehydroascorbate reductase (MDHAR) and dehydroascorbate reductase (DHAR) in tomato (Solanum lycopersicum), an economically important fruit crop. Cytosolic-targeted tomato genes Mdhar and Dhar were cloned and over-expressed under a constitutive promoter in tomato var. Micro-Tom. Lines with increased protein levels and enzymatic activity were identified and examined. Mature green and red ripe fruit from DHAR over-expressing lines had a 1.6 fold increase in AsA content in plants grown under relatively low light conditions (150 μmol m(-2) s(-1)). Conversely, MDHAR over-expressers had significantly reduced AsA levels in mature green fruits by 0.7 fold. Neither over-expressing line had altered levels of AsA in foliar tissues. These results underscore a complex regulation of the AsA pool size in tomato.

  8. Highly Expressive Hakka Art

    Institute of Scientific and Technical Information of China (English)

    JENNIFER LIM

    1996-01-01

    SOUTHERN Jiangxi Province was the birthplace of the Hakka ethnic group and has since been the native home and main transfer hub for the spread of the nationality. The highly expressive art of the Hakkas, including folk songs in Xingguo, colored lantern performances in Shicheng, ancient

  9. Constitutive and heat-shock induced expression of Hsp70 mRNA during chicken testicular development and regression.

    Science.gov (United States)

    Mezquita, B; Mezquita, J; Durfort, M; Mezquita, C

    2001-01-01

    The constitutive and heat shock induced expression of Hsp70 mRNA was investigated in normal adult chicken testis and in adult testis after testicular regression induced by diethylstilbestrol (DES) treatment. In addition to the canonical form of Hsp70 mRNA, we have detected transcripts with an extended 5'UTR and transcripts containing, in the 5'UTR, sequences of the 18S ribosomal RNA. Hsp70 was expressed in unstressed male gonads in adult and regressed testis, being the expression much lower in regressed testis. Upon heat shock at 44 degrees C or 46 degrees C, Hsp70 was highly induced in both tissues. However, when testicular seminiferous tubules were incubated at the chicken internal temperature of 39 degrees C, no induction of Hsp70 was observed in mature testis, while the expression markedly increased in regressed testis. Induction at 39 degrees C was completely inhibited in the presence of 6 mM aspirin. Aspirin in the range 3-10 mM decreases the expression of Hsp70 in unstressed and stressed testicular cells, in striking contrast with the effect observed in other tissues as liver. These data suggest that the expression of Hsp70 is regulated in a specific manner in chicken testis and particularly in the male gonad undergoing regression. Copyright 2001 Wiley-Liss, Inc.

  10. GPR18 undergoes a high degree of constitutive trafficking but is unresponsive to N-Arachidonoyl Glycine

    Directory of Open Access Journals (Sweden)

    David B. Finlay

    2016-03-01

    Full Text Available The orphan receptor GPR18 has become a research target following the discovery of a putative endogenous agonist, N-arachidonoyl glycine (NAGly. Chemical similarity between NAGly and the endocannabinoid anandamide suggested the hypothesis that GPR18 is a third cannabinoid receptor. GPR18-mediated cellular signalling through inhibition of cyclic adenosine monophosphate (cAMP and phosphorylation of extracellular signal-regulated kinase (ERK, in addition to physiological consequences such as regulation of cellular migration and proliferation/apoptosis have been described in response to both NAGly and anandamide. However, discordant findings have also been reported. Here we sought to describe the functional consequences of GPR18 activation in heterologously-expressing HEK cells. GPR18 expression was predominantly intracellular in stably transfected cell lines, but moderate cell surface expression could be achieved in transiently transfected cells which also had higher overall expression. Assays were employed to characterise the ability of NAGly or anandamide to inhibit cAMP or induce ERK phosphorylation through GPR18, or induce receptor trafficking. Positive control experiments, which utilised cells expressing hCB1 receptors (hCB1R, were performed to validate assay design and performance. While these functional pathways in GPR18-expressing cells were not modified on treatment with a panel of putative GPR18 ligands, a constitutive phenotype was discovered for this receptor. Our data reveal that GPR18 undergoes rapid constitutive receptor membrane trafficking—several-fold faster than hCB1R, a highly constitutively active receptor. To enhance the likelihood of detecting agonist-mediated receptor signalling responses, we increased GPR18 protein expression (by tagging with a preprolactin signal sequence and generated a putative constitutively inactive receptor by mutating the hGPR18 gene at amino acid site 108 (alanine to asparagine. This A108N mutant

  11. Constitutive expression of MKS1 confers susceptibility to Botrytis cinerea infection independent of PAD3 expression.

    Science.gov (United States)

    Fiil, Berthe Katrine; Petersen, Morten

    2011-10-01

    Signal transduction through MAPK cascades is essential for eukaryotic cell response to various extracellular stimuli, such as the induction of innate immune responses. Arabidopsis thaliana relies in particular on three of its 20 MAPKs, MPK3,-4,-6, for a proper immune response. Recently we showed that one MPK4-substrate, MKS1, is required for basal resistance against the virulent Pseudomonas syringae and the oomycete Hyaloperonospora arabidopsidis. Overexpression of MKS1 (35S-MKS1) led to increased resistance to the same pathogens but also to an increased susceptibility towards the fungi Botrytis cinerea. MKS1 interacts with the transcription factor WRKY33, which in turn controls the regulation of PAD3 and CYP71A13, two genes, required for proper resistance to B. cinerea. Therefore, we tested if the increased susceptibility towards B. cinerea from 35S-MKS1 was due to deregulation of WRKY33 targets. PAD3 and CYP71A13 expression is similar in 35S-MKS1 and WT after B. cinerea treatment suggesting another mechanism controls 35S-MKS1 susceptibility.

  12. Constitutive expression of transgenes encoding derivatives of the synthetic antimicrobial peptide BP100: impact on rice host plant fitness

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    Nadal Anna

    2012-09-01

    Full Text Available Abstract Background The Biopeptide BP100 is a synthetic and strongly cationic α-helical undecapeptide with high, specific antibacterial activity against economically important plant-pathogenic bacteria, and very low toxicity. It was selected from a library of synthetic peptides, along with other peptides with activities against relevant bacterial and fungal species. Expression of the BP100 series of peptides in plants is of major interest to establish disease-resistant plants and facilitate molecular farming. Specific challenges were the small length, peptide degradation by plant proteases and toxicity to the host plant. Here we approached the expression of the BP100 peptide series in plants using BP100 as a proof-of-concept. Results Our design considered up to three tandemly arranged BP100 units and peptide accumulation in the endoplasmic reticulum (ER, analyzing five BP100 derivatives. The ER retention sequence did not reduce the antimicrobial activity of chemically synthesized BP100 derivatives, making this strategy possible. Transformation with sequences encoding BP100 derivatives (bp100der was over ten-fold less efficient than that of the hygromycin phosphotransferase (hptII transgene. The BP100 direct tandems did not show higher antimicrobial activity than BP100, and genetically modified (GM plants constitutively expressing them were not viable. In contrast, inverted repeats of BP100, whether or not elongated with a portion of a natural antimicrobial peptide (AMP, had higher antimicrobial activity, and fertile GM rice lines constitutively expressing bp100der were produced. These GM lines had increased resistance to the pathogens Dickeya chrysanthemi and Fusarium verticillioides, and tolerance to oxidative stress, with agronomic performance comparable to untransformed lines. Conclusions Constitutive expression of transgenes encoding short cationic α-helical synthetic peptides can have a strong negative impact on rice fitness. However, GM

  13. Constitutive components and induced gene expression are involved in the desiccation tolerance of Selaginella tamariscina.

    Science.gov (United States)

    Liu, Mao-Sen; Chien, Ching-Te; Lin, Tsan-Piao

    2008-04-01

    Selaginella tamariscina, one of the most primitive vascular plants, can remain alive in a desiccated state and resurrect when water becomes available. To evaluate the nature of desiccation tolerance in this plant, we compared the composition of soluble sugars and saturation ratios of phospholipids (PLs) between hydrated and desiccated tissues of S. tamariscina using gas chromatography. In this study, differences in gene expression and ABA contents were also analyzed during dehydration. The results revealed that trehalose (at >130 mg g(-1) DW) was the major soluble sugar, and low saturated fatty acid content in PLs (0.31) was maintained in both hydrated and desiccated tissues. In addition, the ABA content of S. tamariscina increased 3-fold, and genes involved in ABA signaling and cellular protection were up-regulated while photosystem-related genes were down-regulated during dehydration. The biochemical and molecular findings suggest that both constitutive and inducible protective molecules contribute to desiccation tolerance of S. tamariscina.

  14. Development and validation of four Leishmania species constitutively expressing GFP protein. A model for drug discovery and disease pathogenesis studies.

    Science.gov (United States)

    Patel, Asha Parbhu; Deacon, Andrew; Getti, Giulia

    2014-04-01

    Green fluorescent protein (GFP)-parasite transfectants have been widely used as a tool for studying disease pathogenesis in several protozoan models and their application in drug screening assays has increased rapidly. In the past decade, the expression of GFP has been established in several Leishmania species, mostly for in vitro studies. The current work reports generation of four transgenic parasites constitutively expressing GFP (Leishmania mexicana, Leishmania aethiopica, Leishmania tropica and Leishmania major) and their validation as a representative model of infection. This is the first report where stable expression of GFP has been achieved in L. aethiopica and L. tropica. Integration of GFP was accomplished through homologous recombination of the expression construct, pRib1.2αNEOαGFP downstream of the 18S rRNA promoter in all species. A homogeneous and high level expression of GFP was detected in both the promastigote and the intracellular amastigote stages. All transgenic species showed the same growth pattern, ability to infect mammalian host cells and sensitivity to reference drugs as their wild type counterparts. All four transgenic Leishmania are confirmed as models for in vitro and possibly in vivo infections and represent an ideal tool for medium throughput testing of compound libraries.

  15. Constitutive expression of Atlantic salmon Mx1 protein in CHSE-214 cells confers resistance to Infectious Salmon Anaemia virus

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    Kibenge Frederick SB

    2005-08-01

    Full Text Available Abstract Infectious salmon anaemia (ISA is a highly fatal viral disease affecting marine-farmed Atlantic salmon which is caused by ISA virus (ISAV, a fish orthomyxovirus that has recently been assigned to the new genus Isavirus within the family Orthomyxoviridae. Mx proteins are among the interferon (IFN-induced proteins responsible for the development of an antiviral state in vertebrate cells. We used real-time reverse transcription-polymerase chain reaction (RT-PCR and Chinook salmon embryo (CHSE-214 cells constitutively expressing Atlantic salmon Mx1 protein (ASMx1 to examine the antiviral properties of ASMx1 against two ISAV strains, NBISA01 and HKS-36, having phenotypically different growth properties (cytopathic vs non-cytopathic in the CHSE-214 cell line. We present evidence that ISAV is sensitive to ASMx1. CHSE-214 cells constitutively expressing ASMx1 showed increased resistance to infection with the cytopathic ISAV strain NBISA01, manifested as delayed development of cytopathic effects (CPE and significant reduction in the severity of CPE, as well as a 10-fold reduction in virus yield. However, by real-time RT-PCR we observed no significant difference in the mean threshold cycle (Ct values of ISAV RNA levels, suggesting that the ASMx1 activity on ISAV occurs at the post-transcription steps of virus replication, possibly in the cytoplasm.

  16. Constitutive expression of Atlantic salmon Mx1 protein in CHSE-214 cells confers resistance to infectious salmon anaemia virus.

    Science.gov (United States)

    Kibenge, Molly J T; Munir, Khalid; Kibenge, Frederick S B

    2005-08-26

    Infectious salmon anaemia (ISA) is a highly fatal viral disease affecting marine-farmed Atlantic salmon which is caused by ISA virus (ISAV), a fish orthomyxovirus that has recently been assigned to the new genus Isavirus within the family Orthomyxoviridae. Mx proteins are among the interferon (IFN)-induced proteins responsible for the development of an antiviral state in vertebrate cells. We used real-time reverse transcription-polymerase chain reaction (RT-PCR) and Chinook salmon embryo (CHSE-214) cells constitutively expressing Atlantic salmon Mx1 protein (ASMx1) to examine the antiviral properties of ASMx1 against two ISAV strains, NBISA01 and HKS-36, having phenotypically different growth properties (cytopathic vs non-cytopathic) in the CHSE-214 cell line. We present evidence that ISAV is sensitive to ASMx1. CHSE-214 cells constitutively expressing ASMx1 showed increased resistance to infection with the cytopathic ISAV strain NBISA01, manifested as delayed development of cytopathic effects (CPE) and significant reduction in the severity of CPE, as well as a 10-fold reduction in virus yield. However, by real-time RT-PCR we observed no significant difference in the mean threshold cycle (Ct) values of ISAV RNA levels, suggesting that the ASMx1 activity on ISAV occurs at the post-transcription steps of virus replication, possibly in the cytoplasm.

  17. Evaluation of constitutive iron reductase (AtFRO2 expression on mineral accumulation and distribution in soybean (Glycine max. L

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    Marta Wilton Vasconcelos

    2014-04-01

    Full Text Available Iron is an important micronutrient in human and plant nutrition. Adequate iron nutrition during crop production is central for assuring appropriate iron concentrations in the harvestable organs, for human food or animal feed. The whole-plant movement of iron involves several processes, including the reduction of ferric to ferrous iron at several locations throughout the plant, prior to transmembrane trafficking of ferrous iron. In this study, soybean plants that constitutively expressed the AtFRO2 iron reductase gene were analyzed for leaf iron reductase activity, as well as the effect of this transgene's expression on root, leaf, pod wall, and seed mineral concentrations. High Fe supply, in combination with the constitutive expression of AtFRO2, resulted in significantly higher concentrations of different minerals in roots (K, P, Zn, Ca, Ni, Mg and Mo, pod walls (Fe, K, P, Cu and Ni, leaves (Fe, P, Cu, Ca, Ni and Mg and seeds (Fe, Zn, Cu and Ni. Leaf and pod wall iron concentrations increased as much as 500% in transgenic plants, while seed iron concentrations only increased by 10%, suggesting that factors other than leaf and pod wall reductase activity were limiting the translocation of iron to seeds. Protoplasts isolated from transgenic leaves had three-fold higher reductase activity than controls. Expression levels of the iron storage protein, ferritin, were higher in the transgenic leaves than in wild-type, suggesting that the excess iron may be stored as ferritin in the leaves and therefore unavailable for phloem loading and delivery to the seeds. Also, citrate and malate levels in the roots and leaves of transgenic plants were significantly higher than in wild-type, suggesting that organic acid production could be related to the increased accumulation of minerals in roots, leaves and pod walls, but not in the seeds. All together, these results suggest a more ubiquitous role for the iron reductase in whole-plant mineral accumulation and

  18. Constitutive expression of Yes-associated protein (Yap in adult skeletal muscle fibres induces muscle atrophy and myopathy.

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    Robert N Judson

    Full Text Available The aim of this study was to investigate the function of the Hippo pathway member Yes-associated protein (Yap, gene name Yap1 in skeletal muscle fibres in vivo. Specifically we bred an inducible, skeletal muscle fibre-specific knock-in mouse model (MCK-tTA-hYAP1 S127A to test whether the over expression of constitutively active Yap (hYAP1 S127A is sufficient to drive muscle hypertrophy or stimulate changes in fibre type composition. Unexpectedly, after 5-7 weeks of constitutive hYAP1 S127A over expression, mice suddenly and rapidly lost 20-25% body weight and suffered from gait impairments and kyphosis. Skeletal muscles atrophied by 34-40% and the muscle fibre cross sectional area decreased by ≈40% when compared to control mice. Histological analysis revealed evidence of skeletal muscle degeneration and regeneration, necrotic fibres and a NADH-TR staining resembling centronuclear myopathy. In agreement with the histology, mRNA expression of markers of regenerative myogenesis (embryonic myosin heavy chain, Myf5, myogenin, Pax7 and muscle protein degradation (atrogin-1, MuRF1 were significantly elevated in muscles from transgenic mice versus control. No significant changes in fibre type composition were detected using ATPase staining. The phenotype was largely reversible, as a cessation of hYAP1 S127A expression rescued body and muscle weight, restored muscle morphology and prevented further pathological progression. To conclude, high Yap activity in muscle fibres does not induce fibre hypertrophy nor fibre type changes but instead results in a reversible atrophy and deterioration.

  19. Constitutive Expression of the Vi Polysaccharide Capsular Antigen in Attenuated Salmonella enterica Serovar Typhi Oral Vaccine Strain CVD 909

    Science.gov (United States)

    Wang, Jin Yuan; Noriega, Fernando R.; Galen, James E.; Barry, Eileen; Levine, Myron M.

    2000-01-01

    Live oral Ty21a and parenteral Vi polysaccharide vaccines provide significant protection against typhoid fever, albeit by distinct immune mechanisms. Vi stimulates serum immunoglobulin G Vi antibodies, whereas Ty21a, which does not express Vi, elicits humoral and cell-mediated immune responses other than Vi antibodies. Protection may be enhanced if serum Vi antibody as well as cell-mediated and humoral responses can be stimulated. Disappointingly, several new attenuated Salmonella enterica serovar Typhi oral vaccines (e.g., CVD 908-htrA and Ty800) that elicit serum O and H antibody and cell-mediated responses following a single dose do not stimulate serum Vi antibody. Vi expression is regulated in response to environmental signals such as osmolarity by controlling the transcription of tviA in the viaB locus. To investigate if Vi antibodies can be stimulated if Vi expression is rendered constitutive, we replaced PtviA in serovar Typhi vaccine CVD 908-htrA with the constitutive promoter Ptac, resulting in CVD 909. CVD 909 expresses Vi even under high-osmolarity conditions and is less invasive for Henle 407 cells. In mice immunized with a single intranasal dose, CVD 909 was more immunogenic than CVD 908-htrA in eliciting serum Vi antibodies (geometric mean titer of 160 versus 49, P = 0.0007), whereas O antibody responses were virtually identical (geometric mean titer of 87 versus 80). In mice challenged intraperitoneally with wild-type serovar Typhi 4 weeks after a single intranasal immunization, the mortality of those immunized with CVD 909 (3 of 8) was significantly lower than that of control mice (10 of 10, P = 0.043) or mice given CVD 908-htrA (9 of 10, P = 0.0065). PMID:10899868

  20. Evaluation of constitutive iron reductase (AtFRO2) expression on mineral accumulation and distribution in soybean (Glycine max. L).

    Science.gov (United States)

    Vasconcelos, Marta W; Clemente, Thomas E; Grusak, Michael A

    2014-01-01

    Iron is an important micronutrient in human and plant nutrition. Adequate iron nutrition during crop production is central for assuring appropriate iron concentrations in the harvestable organs, for human food or animal feed. The whole-plant movement of iron involves several processes, including the reduction of ferric to ferrous iron at several locations throughout the plant, prior to transmembrane trafficking of ferrous iron. In this study, soybean plants that constitutively expressed the AtFRO2 iron reductase gene were analyzed for leaf iron reductase activity, as well as the effect of this transgene's expression on root, leaf, pod wall, and seed mineral concentrations. High Fe supply, in combination with the constitutive expression of AtFRO2, resulted in significantly higher concentrations of different minerals in roots (K, P, Zn, Ca, Ni, Mg, and Mo), pod walls (Fe, K, P, Cu, and Ni), leaves (Fe, P, Cu, Ca, Ni, and Mg) and seeds (Fe, Zn, Cu, and Ni). Leaf and pod wall iron concentrations increased as much as 500% in transgenic plants, while seed iron concentrations only increased by 10%, suggesting that factors other than leaf and pod wall reductase activity were limiting the translocation of iron to seeds. Protoplasts isolated from transgenic leaves had three-fold higher reductase activity than controls. Expression levels of the iron storage protein, ferritin, were higher in the transgenic leaves than in wild-type, suggesting that the excess iron may be stored as ferritin in the leaves and therefore unavailable for phloem loading and delivery to the seeds. Also, citrate and malate levels in the roots and leaves of transgenic plants were significantly higher than in wild-type, suggesting that organic acid production could be related to the increased accumulation of minerals in roots, leaves, and pod walls, but not in the seeds. All together, these results suggest a more ubiquitous role for the iron reductase in whole-plant mineral accumulation and distribution.

  1. Constitutive expression of the Period1 gene impairs behavioral and molecular circadian rhythms.

    Science.gov (United States)

    Numano, Rika; Yamazaki, Shin; Umeda, Nanae; Samura, Tomonori; Sujino, Mitsugu; Takahashi, Ri-ichi; Ueda, Masatsugu; Mori, Akiko; Yamada, Kazunori; Sakaki, Yoshiyuki; Inouye, Shin-ichi T; Menaker, Michael; Tei, Hajime

    2006-03-07

    Three mammalian Period (Per) genes, termed Per1, Per2, and Per3, have been identified as structural homologues of the Drosophila circadian clock gene, period (per). The three Per genes are rhythmically expressed in the suprachiasmatic nucleus (SCN), the central circadian pacemaker in mammals. The phases of peak mRNA levels for the three Per genes in the SCN are slightly different. Light sequentially induces the transcripts of Per1 and Per2 but not of Per3 in mice. These data and others suggest that each Per gene has a different but partially redundant function in mammals. To elucidate the function of Per1 in the circadian system in vivo, we generated two transgenic rat lines in which the mouse Per1 (mPer1) transcript was constitutively expressed under the control of either the human elongation factor-1alpha (EF-1alpha) or the rat neuron-specific enolase (NSE) promoter. The transgenic rats exhibited an approximately 0.6-1.0-h longer circadian period than their wild-type siblings in both activity and body temperature rhythms. Entrainment in response to light cycles was dramatically impaired in the transgenic rats. Molecular analysis revealed that the amplitudes of oscillation in the rat Per1 (rPer1) and rat Per2 (rPer2) mRNAs were significantly attenuated in the SCN and eyes of the transgenic rats. These results indicate that either the level of Per1, which is raised by overexpression, or its rhythmic expression, which is damped or abolished in over expressing animals, is critical for normal entrainment of behavior and molecular oscillation of other clock genes.

  2. Establishment of Sf9 transformants constitutively expressing PBAN receptor (PBANR variants: application to functional evaluation

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    Jae Min Lee

    2012-04-01

    Full Text Available To facilitate further evaluation of pheromone biosynthesis activating neuropeptide receptor (PBANR functionality and regulation, we generated cultured insect cell lines constitutively expressing green fluorescent protein chimeras of the recently identified Bombyx mori PBANR (BommoPBANR and Pseudaletia separata PBANR (PsesePBANR variants. Fluorescent chimeras included the BommoPBANR-A, B, and C variants and the PsesePBANR-B and C variants. Cell lines expressing non-chimeric BommoPBANR-B and C variants were also generated. Functional evaluation of these transformed cell lines using confocal laser microscopy revealed that a Rhodamine Red-labeled PBAN derivative (RR-C10PBANR2K specifically co-localized with all of the respective PBANR variants at the plasma membrane. Near complete internalization of the fluorescent RR-C10PBANR2K ligand 30 min after binding was observed in all cell lines except those expressing the BommoPBANR-A variant, in which the ligand/receptor complex remained at the plasma membrane. Fluorescent Ca2+ imaging further showed that, unlike the BommoPBANR-B or BommoPBANR-C cell lines, RR-C10PBANR2K binding failed to mobilize extracellular Ca2+ in the BommoPBANR-A cell line even at concentrations of 10 M. These observations demonstrate a clear functional difference between the BommoPBANR-A variant and the BommoPBANR-B and –C variants in terms of receptor regulation and activation of downstream effector molecules. We also found that, contrary to previous reports, ligand-induced internalization of BommoPBANR-B and BommoPBANR-C in cell lines stably expressing these variants occurred in the absence of extracellular Ca2+.

  3. Enhancement of endothelial cell migration by constitutively active LPA{sub 1}-expressing tumor cells

    Energy Technology Data Exchange (ETDEWEB)

    Kitayoshi, Misaho; Kato, Kohei; Tanabe, Eriko; Yoshikawa, Kyohei; Fukui, Rie [Division of Cancer Biology and Bioinformatics, Department of Life Science, Faculty of Science and Engineering, Kinki University, 3-4-1, Kowakae, Higashiosaka, Osaka 577-8502 (Japan); Fukushima, Nobuyuki [Division of Molecular Neurobiology, Department of Life Science, Faculty of Science and Engineering, Kinki University, 3-4-1, Kowakae, Higashiosaka, Osaka 577-8502 (Japan); Tsujiuchi, Toshifumi, E-mail: ttujiuch@life.kindai.ac.jp [Division of Cancer Biology and Bioinformatics, Department of Life Science, Faculty of Science and Engineering, Kinki University, 3-4-1, Kowakae, Higashiosaka, Osaka 577-8502 (Japan)

    2012-06-01

    Highlights: Black-Right-Pointing-Pointer Mutated LPA{sub 1} stimulates cell migration of endothelial cells. Black-Right-Pointing-Pointer VEGF expressions are increased by mutated LPA{sub 1}. Black-Right-Pointing-Pointer LPA signaling via mutated LPA{sub 1} is involved in angiogenesis. Black-Right-Pointing-Pointer Mutated LPA{sub 1} promotes cancer cell progression. -- Abstract: Lysophosphatidic acid (LPA) receptors belong to G protein-coupled transmembrane receptors (LPA receptors; LPA{sub 1} to LPA{sub 6}). They indicate a variety of cellular response by the interaction with LPA, including cell proliferation, migration and differentiation. Recently, we have reported that constitutive active mutated LPA{sub 1} induced the strong biological effects of rat neuroblastoma B103 cells. In the present study, we examined the effects of mutated LPA{sub 1} on the interaction between B103 cells and endothelial F-2 cells. Each LPA receptor expressing B103 cells were maintained in serum-free DMEM and cell motility assay was performed with a Cell Culture Insert. When F-2 cells were cultured with conditioned medium from Lpar1 and Lpar3-expressing cells, the cell motility of F-2 cells was significantly higher than control cells. Interestingly, the motile activity of F-2 cells was strongly induced by mutated LPA{sub 1} than other cells, correlating with the expression levels of vascular endothelial growth factor (Vegf)-A and Vegf-C. Pretreatment of LPA signaling inhibitors inhibited F-2 cell motility stimulated by mutated LPA{sub 1}. These results suggest that activation of LPA signaling via mutated LPA{sub 1} may play an important role in the promotion of angiogenesis in rat neuroblastoma cells.

  4. Constitutive expression of pathogenesis-related proteins PR-1, GRP, and PR-S in tobacco has no effect on virus infection.

    Science.gov (United States)

    Linthorst, H J; Meuwissen, R L; Kauffmann, S; Bol, J F

    1989-03-01

    Samsun NN tobacco cells were transformed with chimeric genes for pathogenesis-related (PR) proteins derived from genomic (PR-1a, GRP) or cDNA (PR-S) clones under the transcriptional control of the cauliflower mosaic virus 35S promoter. Regenerated plants were assayed by RNA and protein gel blotting, and plants showing high specific expression of the inserted genes were selected for self-pollination and seed formation. Inspection of second generation transformants showed that constitutive expression of PR-1a, GRP, and PR-S in tobacco in general does not have an effect on the phenotypic appearance of the plants or the expression of other endogenous PR genes. Furthermore, constitutive expression of the above genes does not affect the susceptibility of the plants to infection with tobacco mosaic virus or alfalfa mosaic virus.

  5. Constitutive knox1 gene expression in dandelion (Taraxacum officinale, Web.) changes leaf morphology from simple to compound.

    Science.gov (United States)

    Müller, Kai J; He, Xinqiang; Fischer, Rainer; Prüfer, Dirk

    2006-10-01

    Seed plants with compound leaves constitute a polyphyletic group, but studies of diverse taxa show that genes of the class 1 KNOTTED-LIKE HOMEOBOX (KNOX1) family are often involved in compound leaf development. This suggests that knox1 genes have been recruited on multiple occasions during angiosperm evolution (Bharathan et al. in Science 296:1858-1860, 2002). In agreement with this, we demonstrate that the simple leaf of dandelion (Taraxacum officinale Web.) can be converted into a compound leaf by the constitutive expression of heterologous knox1 genes. Dandelion is a rosette plant of the family Asteraceae, characterised by simple leaves with deeply lobed margins and endogenous knox1 gene expression. Transgenic dandelion plants constitutively expressing the barley (Hordeum vulgare L.) hooded gene (bkn3, barley knox3) or the related bkn1 gene, developed compound leaves featuring epiphyllous rosettes. We discuss these results in the context of two current models of compound leaf formation.

  6. Identification of chemical modulators of the constitutive activated receptor (CAR) in a gene expression compendium.

    Science.gov (United States)

    Oshida, Keiyu; Vasani, Naresh; Jones, Carlton; Moore, Tanya; Hester, Susan; Nesnow, Stephen; Auerbach, Scott; Geter, David R; Aleksunes, Lauren M; Thomas, Russell S; Applegate, Dawn; Klaassen, Curtis D; Corton, J Christopher

    2015-01-01

    The nuclear receptor family member constitutive activated receptor (CAR) is activated by structurally diverse drugs and environmentally-relevant chemicals leading to transcriptional regulation of genes involved in xenobiotic metabolism and transport. Chronic activation of CAR increases liver cancer incidence in rodents, whereas suppression of CAR can lead to steatosis and insulin insensitivity. Here, analytical methods were developed to screen for chemical treatments in a gene expression compendium that lead to alteration of CAR activity. A gene expression biomarker signature of 83 CAR-dependent genes was identified using microarray profiles from the livers of wild-type and CAR-null mice after exposure to three structurally-diverse CAR activators (CITCO, phenobarbital, TCPOBOP). A rank-based algorithm (Running Fisher's algorithm (p-value ≤ 10(-4))) was used to evaluate the similarity between the CAR biomarker signature and a test set of 28 and 32 comparisons positive or negative, respectively, for CAR activation; the test resulted in a balanced accuracy of 97%. The biomarker signature was used to identify chemicals that activate or suppress CAR in an annotated mouse liver/primary hepatocyte gene expression database of ~1850 comparisons. CAR was activated by 1) activators of the aryl hydrocarbon receptor (AhR) in wild-type but not AhR-null mice, 2) pregnane X receptor (PXR) activators in wild-type and to lesser extents in PXR-null mice, and 3) activators of PPARα in wild-type and PPARα-null mice. CAR was consistently activated by five conazole fungicides and four perfluorinated compounds. Comparison of effects in wild-type and CAR-null mice showed that the fungicide propiconazole increased liver weight and hepatocyte proliferation in a CAR-dependent manner, whereas the perfluorinated compound perfluorooctanoic acid (PFOA) increased these endpoints in a CAR-independent manner. A number of compounds suppressed CAR coincident with increases in markers of

  7. Identification of centrarchid hepcidins and evidence that 17β-estradiol disrupts constitutive expression of hepcidin-1 and inducible expression of hepcidin-2 in largemouth bass (Micropterus salmoides)

    Science.gov (United States)

    Robertson, L.S.; Iwanowicz, L.R.; Marranca, J.M.

    2009-01-01

    Hepcidin is a highly conserved antimicrobial peptide and iron-regulatory hormone. Here, we identify two hepcidin genes (hep-1 and hep-2) in largemouth bass (Micropterus salmoides) and smallmouth bass (Micropterus dolomieu). Hepcidin-1 contains a putative ATCUN metal-binding site in the amino-terminus that is missing in hepcidin-2, suggesting that hepcidin-1 may function as an iron-regulatory hormone. Both hepcidins are predominately expressed in the liver of largemouth bass, similar to other fish and mammals. Experimental exposure of pond-raised largemouth bass to 17β-estradiol and/or the bacteria Edwardsiella ictaluri led to distinct changes in expression of hep-1 and hep-2. Estradiol reduced the constitutive expression of hep-1 in the liver. Bacterial exposure induced expression of hep-2, suggesting that hepcidin-2 may have an antimicrobial function, and this induction was abolished by estradiol. To our knowledge, this is the first report of the regulation of hepcidin expression by estradiol in either fish or mammals.

  8. Constitutive expression of tdTomato protein as a cytotoxicity and proliferation marker for space radiation biology.

    Science.gov (United States)

    Chishti, Arif A; Hellweg, Christine E; Berger, Thomas; Baumstark-Khan, Christa; Feles, Sebastian; Kätzel, Thorben; Reitz, Günther

    2015-01-01

    The radiation risk assessment for long-term space missions requires knowledge on the biological effectiveness of different space radiation components, e.g. heavy ions, on the interaction of radiation and other space environmental factors such as microgravity, and on the physical and biological dose distribution in the human body. Space experiments and ground-based experiments at heavy ion accelerators require fast and reliable test systems with an easy readout for different endpoints. In order to determine the effect of different radiation qualities on cellular proliferation and the biological depth dose distribution after heavy ion exposure, a stable human cell line expressing a novel fluorescent protein was established and characterized. tdTomato, a red fluorescent protein of the new generation with fast maturation and high fluorescence intensity, was selected as reporter of cell proliferation. Human embryonic kidney (HEK/293) cells were stably transfected with a plasmid encoding tdTomato under the control of the constitutively active cytomegalovirus (CMV) promoter (ptdTomato-N1). The stably transfected cell line was named HEK-ptdTomato-N1 8. This cytotoxicity biosensor was tested by ionizing radiation (X-rays and accelerated heavy ions) exposure. As biological endpoints, the proliferation kinetics and the cell density reached 100 h after irradiation reflected by constitutive expression of the tdTomato were investigated. Both were reduced dose-dependently after radiation exposure. Finally, the cell line was used for biological weighting of heavy ions of different linear energy transfer (LET) as space-relevant radiation quality. The relative biological effectiveness of accelerated heavy ions in reducing cellular proliferation peaked at an LET of 91 keV/μm. The results of this study demonstrate that the HEK-ptdTomato-N1 reporter cell line can be used as a fast and reliable biosensor system for detection of cytotoxic damage caused by ionizing radiation.

  9. Constitutively Expressed IFITM3 Protein in Human Endothelial Cells Poses an Early Infection Block to Human Influenza Viruses.

    Science.gov (United States)

    Sun, Xiangjie; Zeng, Hui; Kumar, Amrita; Belser, Jessica A; Maines, Taronna R; Tumpey, Terrence M

    2016-12-15

    A role for pulmonary endothelial cells in the orchestration of cytokine production and leukocyte recruitment during influenza virus infection, leading to severe lung damage, has been recently identified. As the mechanistic pathway for this ability is not fully known, we extended previous studies on influenza virus tropism in cultured human pulmonary endothelial cells. We found that a subset of avian influenza viruses, including potentially pandemic H5N1, H7N9, and H9N2 viruses, could infect human pulmonary endothelial cells (HULEC) with high efficiency compared to human H1N1 or H3N2 viruses. In HULEC, human influenza viruses were capable of binding to host cellular receptors, becoming internalized and initiating hemifusion but failing to uncoat the viral nucleocapsid and to replicate in host nuclei. Unlike numerous cell types, including epithelial cells, we found that pulmonary endothelial cells constitutively express a high level of the restriction protein IFITM3 in endosomal compartments. IFITM3 knockdown by small interfering RNA (siRNA) could partially rescue H1N1 virus infection in HULEC, suggesting IFITM3 proteins were involved in blocking human influenza virus infection in endothelial cells. In contrast, selected avian influenza viruses were able to escape IFITM3 restriction in endothelial cells, possibly by fusing in early endosomes at higher pH or by other, unknown mechanisms. Collectively, our study demonstrates that the human pulmonary endothelium possesses intrinsic immunity to human influenza viruses, in part due to the constitutive expression of IFITM3 proteins. Notably, certain avian influenza viruses have evolved to escape this restriction, possibly contributing to virus-induced pneumonia and severe lung disease in humans. Avian influenza viruses, including H5N1 and H7N9, have been associated with severe respiratory disease and fatal outcomes in humans. Although acute respiratory distress syndrome (ARDS) and progressive pulmonary endothelial damage

  10. Differential Constitutive and Cytokine-Modulated Expression of Human Toll-like Receptors in Primary Neutrophils, Monocytes, and Macrophages

    Directory of Open Access Journals (Sweden)

    D. Shane O'Mahony, Uyenvy Pham, Ramesh Iyer, Thomas R. Hawn, W. Conrad Liles

    2008-01-01

    Full Text Available Human Toll-like receptors (TLRs comprise a family of proteins that recognizes pathogen-associated molecular patterns (PAMPs and initiates host innate immune responses. Neutrophils, monocytes, and macrophages are critical cellular components of the human innate immune system. Proinflammatory cytokines, such as granulocyte colony-stimulating factor (G-CSF, granulocyte-macrophage colony-stimulating factor (GM-CSF, macrophage colony-stimulating factor (M-CSF, and interferon-γ (IFN-γ, have been shown to up-regulate microbicidal activity in these effector cells of innate immunity. Currently, the cellular and molecular mechanisms responsible for these effects are not completely understood. We hypothesized that these cytokines may up-regulate TLR expression as a mechanism to facilitate microbial recognition and augment the innate immune response. Using quantitative realtime rt-PCR technology, we examined constitutive expression of TLR2, TLR4, TLR5, and TLR9 mRNA and the effects of G-CSF, GM-CSF, M-CSF, and IFN-γ on TLR mRNA expression in purified populations of normal human neutrophils, monocytes, and monocyte-derived macrophages. Relative constitutive expression of TLR2, TLR4, and TLR9 was similar in neutrophils and monocytes. Constitutive expression of TLR5 was less in neutrophils compared to monocytes. Constitutive expression of TLR4 was greater and that of TLR9 lower in monocyte-derived macrophages compared to monocytes. Of the cytokines examined, IFN-γ and GM-CSF caused the greatest effects on TLR expression. IFN- γ up-regulated TLR2 and TLR4 in neutrophils and monocytes. GM-CSF up-regulated expression of TLR2 and TLR4 in neutrophils and TLR2 in monocytes. TLR5 was down-regulated by inflammatory cytokines in monocytes. These results suggest a potential role for IFN- γ and/or GM-CSF as therapeutic immunomodulators of the host defense to infection.

  11. Constitutive expression of pluripotency-associated genes in mesodermal progenitor cells (MPCs.

    Directory of Open Access Journals (Sweden)

    Simone Pacini

    Full Text Available BACKGROUND: We recently characterized a progenitor of mesodermal lineage (MPCs from the human bone marrow of adults or umbilical cord blood. These cells are progenitors able to differentiate toward mesenchymal, endothelial and cardiomyogenic lineages. Here we present an extensive molecular characterization of MPCs, from bone marrow samples, including 39 genes involved in stem cell machinery, differentiation and cell cycle regulation. METHODOLOGY/PRINCIPAL FINDINGS: MPCs are cytofluorimetrically characterized and quantitative RT-PCR was performed to evaluate the gene expression profile, comparing it with MSCs and hESCs lines. Immunofluorescence and dot-blot analysis confirm qRT-PCR data. MPCs exhibit an increased expression of OCT4, NANOG, SALL4, FBX15, SPP1 and to a lesser extent c-MYC and KLF4, but lack LIN28 and SOX2. MPCs highly express SOX15. CONCLUSIONS/SIGNIFICANCE: MPCs express many pluripotency-associated genes and show a peculiar Oct-4 molecular circuit. Understanding this unique molecular mechanism could lead to identifying MPCs as feasible, long telomeres, target cells for reprogramming with no up-regulation of the p53 pathway. Furthermore MPCs are easily and inexpensively harvested from human bone marrow.

  12. Cestrum yellow leaf curling virus (CmYLCV) promoter: a new strong constitutive promoter for heterologous gene expression in a wide variety of crops.

    Science.gov (United States)

    Stavolone, Livia; Kononova, Maria; Pauli, Sandra; Ragozzino, Antonio; de Haan, Peter; Milligan, Steve; Lawton, Kay; Hohn, Thomas

    2003-11-01

    Appropriately regulated gene expression requires a suitable promoter. A number of promoters have been isolated and shown to be functional in plants, but only a few of them activate transcription of transgenes at high levels constitutively. We report here the cloning and characterization of a novel, constitutively expressed promoter isolated from Cestrum yellow leaf curling virus (CmYLCV), a double-stranded DNA plant pararetrovirus belonging to the Caulimoviridae family. The CmYLCV promoter is highly active in callus, meristems and vegetative and reproductive tissues in Arabidopsis thaliana, Nicotiana tabacum, Lycopersicon esculentum, Zea mays and Oryza sativa. Furthermore, the level of expression is comparable to, or higher than, that from the CaMV 35S, the 'super-promoter' or the maize ubiquitin 1 promoters, three frequently used promoters in agricultural biotechnology. The heritable, strong and constitutive activity in both monocotyledonous and dicotyledonous plants, combined with the extremely narrow CmYLCV host range, makes the CmYLCV promoter an attractive tool for regulating transgene expression in a wide variety of plant species.

  13. Conserved but Attenuated Parental Gene Expression in Allopolyploids: Constitutive Zinc Hyperaccumulation in the Allotetraploid Arabidopsis kamchatica.

    Science.gov (United States)

    Paape, Timothy; Hatakeyama, Masaomi; Shimizu-Inatsugi, Rie; Cereghetti, Teo; Onda, Yoshihiko; Kenta, Tanaka; Sese, Jun; Shimizu, Kentaro K

    2016-11-01

    Allopolyploidization combines parental genomes and often confers broader species distribution. However, little is known about parentally transmitted gene expression underlying quantitative traits following allopolyploidization because of the complexity of polyploid genomes. The allopolyploid species Arabidopsis kamchatica is a natural hybrid of the zinc hyperaccumulator Arabidopsis halleri and of the nonaccumulator Arabidopsis lyrata We found that A. kamchatica retained the ability to hyperaccumulate zinc from A. halleri and grows in soils with both low and high metal content. Hyperaccumulation of zinc by A. kamchatica was reduced to about half of A. halleri, but is 10-fold greater than A. lyrata Homeologs derived from A. halleri had significantly higher levels of expression of genes such as HEAVY METAL ATPASE4 (HMA4), METAL TRANSPORTER PROTEIN1 and other metal ion transporters than those derived from A. lyrata, which suggests cis-regulatory differences. A. kamchatica has on average about half the expression of these genes compared with A. halleri due to fixed heterozygosity inherent in allopolyploids. Zinc treatment significantly changed the ratios of expression of 1% of homeologous pairs, including genes putatively involved in metal homeostasis. Resequencing data showed a significant reduction in genetic diversity over a large genomic region (290 kb) surrounding the HMA4 locus derived from the A. halleri parent compared with the syntenic A. lyrata-derived region, which suggests different evolutionary histories. We also estimated that three A. halleri-derived HMA4 copies are present in A. kamchatica Our findings support a transcriptomic model in which environment-related transcriptional patterns of both parents are conserved but attenuated in the allopolyploids. © The Author 2016. Published by Oxford University Press on behalf of the Society for Molecular Biology and Evolution.

  14. Field tolerance to fungal pathogens of Brassica napus constitutively expressing a chimeric chitinase gene

    Energy Technology Data Exchange (ETDEWEB)

    Grison, R.; Grezes-Besset, B.; Lucante, N. [Rustica Prograin Genetique, Mondonville (France)] [and others

    1996-05-01

    Constitutive overexpression of a protein involved in plant defense mechanisms to disease is one of the strategies proposed to increase plant tolerance to fungal pathogens. A hybrid endochitinase gene under a constitutive promoter was introduced by Agrobacterium-mediated transformation into a winter-type oilseed rape (Brassica napus var. oleifera) inbred line. Progeny from transformed plants was challenged using three different fungal pathogens (Cylindrosporium concentricum, Phoma lingam, Sclerotinia sclerotiorum) in field trials at two different geographical locations. These plants exhibited an increased tolerance to disease as compared with the nontransgenic parental plants. 31 refs., 1 fig., 2 tabs.

  15. Aggravation of psoriasis by infections: a constitutional trait or a variable expression?

    NARCIS (Netherlands)

    Blok, S.; Vissers, W.H.P.M.; Duijnhoven, M. van; Kerkhof, P.C.M. van de

    2004-01-01

    Focal infections are a well known triggering factor in patients with psoriasis. A question which remains to be answered is whether relapsing of psoriasis after infections is a constitutional trait of a restricted population or whether it is a feature of all patients in some phase of their disease. A

  16. Aggravation of psoriasis by infections: a constitutional trait or a variable expression?

    NARCIS (Netherlands)

    Blok, S.; Vissers, W.H.P.M.; Duijnhoven, M. van; Kerkhof, P.C.M. van de

    2004-01-01

    Focal infections are a well known triggering factor in patients with psoriasis. A question which remains to be answered is whether relapsing of psoriasis after infections is a constitutional trait of a restricted population or whether it is a feature of all patients in some phase of their disease. A

  17. Cisplatin hypersensitivity of testicular germ cell tumors is determined by high constitutive Noxa levels mediated by Oct-4.

    Science.gov (United States)

    Gutekunst, Matthias; Mueller, Thomas; Weilbacher, Andrea; Dengler, Michael A; Bedke, Jens; Kruck, Stephan; Oren, Moshe; Aulitzky, Walter E; van der Kuip, Heiko

    2013-03-01

    Testicular germ cell tumors (TGCT) are considered a paradigm of chemosensitive tumors. Embryonal carcinoma cells represent the pluripotent entity of TGCTs and are characterized by expression of Oct-4, a key regulator of pluripotency and a determinant of their inherent hypersensitivity to cisplatin. However, the mechanisms underlying this Oct-4-mediated sensitivity are poorly understood. We previously showed that p53 is a major player in cisplatin hypersensitivity and therefore investigated whether Oct-4 may directly affect p53 activity. Despite a significant decrease in sensitivity, depletion of Oct-4 neither did alter cisplatin-induced transactivation of p53 target genes nor its subcellular localization. These data indicate that, rather than directly modulating p53 activity, Oct-4 provides a cellular context that augments the proapoptotic activity of p53. As mitochondrial priming by the Bcl-2 family is a known determinant of chemosensitivity, we compared the constitutive levels of these proteins in Oct-4-positive and -depleted cells. We identified Noxa as the only Bcl-2 family protein to be highly correlated with Oct-4 status and cisplatin sensitivity. Compared with differentiated cells, constitutive Noxa levels were significantly higher in Oct-4-positive cell lines and cancer patient samples. Furthermore, RNA interference-mediated knockdown of Oct-4 resulted in reduced Noxa transcript, in an almost complete loss of constitutive Noxa protein and decreased cisplatin hypersensitivity to a similar extent as did Noxa depletion. In conclusion, our study indicates that Noxa is a central determinant of hypersensitivity to cisplatin. Oct-4-dependent high constitutive levels of this BH3-only protein prime embryonal carcinoma cells to undergo rapid and massive apoptosis in response to p53 activation.

  18. Constitutive arsenite oxidase expression detected in arsenic-hypertolerant Pseudomonas xanthomarina S11.

    Science.gov (United States)

    Koechler, Sandrine; Arsène-Ploetze, Florence; Brochier-Armanet, Céline; Goulhen-Chollet, Florence; Heinrich-Salmeron, Audrey; Jost, Bernard; Lièvremont, Didier; Philipps, Muriel; Plewniak, Frédéric; Bertin, Philippe N; Lett, Marie-Claire

    2015-04-01

    Pseudomonas xanthomarina S11 is an arsenite-oxidizing bacterium isolated from an arsenic-contaminated former gold mine in Salsigne, France. This bacterium showed high resistance to arsenite and was able to oxidize arsenite to arsenate at concentrations up to 42.72 mM As[III]. The genome of this strain was sequenced and revealed the presence of three ars clusters. One of them is located on a plasmid and is organized as an "arsenic island" harbouring an aio operon and genes involved in phosphorous metabolism, in addition to the ars genes. Neither the aioXRS genes nor a specific sigma-54-dependent promoter located upstream of aioBA genes, both involved in regulation of arsenite oxidase expression in other arsenite-oxidizing bacteria, could be identified in the genome. This observation is in accordance with the fact that no difference was observed in expression of arsenite oxidase in P. xanthomarina S11, whether or not the strain was grown in the presence of As[III].

  19. Antigen-specific monoclonal antibodies isolated from B cells expressing constitutively active STAT5.

    Directory of Open Access Journals (Sweden)

    Ferenc A Scheeren

    Full Text Available BACKGROUND: Fully human monoclonal antibodies directed against specific pathogens have a high therapeutic potential, but are difficult to generate. METHODOLOGY/PRINCIPAL FINDINGS: Memory B cells were immortalized by expressing an inducible active mutant of the transcription factor Signal Transducer and Activator of Transcription 5 (STAT5. Active STAT5 inhibits the differentiation of B cells while increasing their replicative life span. We obtained cloned B cell lines, which produced antibodies in the presence of interleukin 21 after turning off STAT5. We used this method to obtain monoclonal antibodies against the model antigen tetanus toxin. CONCLUSIONS/SIGNIFICANCE: Here we describe a novel and relatively simple method of immortalizing antigen-specific human B cells for isolation of human monoclonal antibodies. These results show that STAT5 overexpression can be employed to isolate antigen specific antibodies from human memory B cells.

  20. Expression profiling of constitutive mast cells reveals a unique identity within the immune system

    Science.gov (United States)

    Dwyer, Daniel F.; Barrett, Nora A.; Austen, K. Frank

    2016-01-01

    Mast cells are evolutionarily ancient sentinel cells. Like basophils, mast cells express the high-affinity IgE receptor and are implicated in host defense and diverse immune-mediated diseases. To better characterize the function of these cells, we assessed the transcriptional profiles of mast cells isolated from peripheral connective tissues and basophils isolated from spleen and blood. We found that mast cells were transcriptionally distinct, clustering independently from all other profiled cells, and that mast cells demonstrated considerably greater heterogeneity across tissues than previously appreciated. We observed minimal homology between mast cells and basophils, which share more overlap with other circulating granulocytes than with mast cells. Derivation of mast cell and basophil transcriptional signatures underscores their differential capacity to detect environmental signals and influence the inflammatory milieu. PMID:27135604

  1. CONSTITUTIVE COMPUTATIONAL MODELLING FOUNDATION OF PIEZOELECTRONIC MICROSTRUCTURES AND APPLICATION TO HIGH-FREQUENCY MICROCHIP DSAW RESONATORS

    Institute of Scientific and Technical Information of China (English)

    张武; 唐锦春

    2002-01-01

    This paper establishes a piezoelectric constitutive computational approach based on generalized eigenvalue and multivariable finite element solutions with potential applications to accurate and effective analysis of layered piezoelectric microstructures of arbitrary geometries and different anisotropic materials, to ease the limitation of current computer capacity in analyzing large-scale high-frequency disturbed surface acoustic waves (DSAW) by mounted electrodes in piezoelectric devices such as microchip SAW resonators. A new incompatible generalized hybrid/mixed element GQM5 is also proposed for improving predictions of the piezoelectric surface mount thermal stresses that are shear-dominated. The (generalized) plane strain constitutive model is numerically verified for piezoelectric finite element computation. With the help of computational piezoelectricity (electro-mechanics) for general layered structures with metal electrodes and anisotropic piezoelectric substrates, some new interesting, reliable and fundamental constitutive finite element results are obtained for high-frequency piezoelectric and mechanical SAW propagations and can be used for further applications. The ST-cut FEA results agree quite well with available exact and lab solutions for free surface case.

  2. Identification and comparison of different constitutive laws for high speed solicitation

    Science.gov (United States)

    Sattouf, C.; Dalverny, O.; Rakotomalala, R.

    2003-09-01

    This paper deals with a methodology for the identification of constitutive laws for metallic materials under dynamic loading at high strain rates. Different constitutive elastoplastic behaviors are compared to represent several materials such as 42CD4 steel or aluminum alloy A2017T3. The classical radial return algorithm is used to integrate the elastoplasticity law with a mixed isotropic/kinematic hardening. The theory has been used to implement the constitutive law in Abaqus/Explicit Vumat subroutine. In the second part we present the different types of elastoplastic hardening laws such as Johnson-Cook, modified Johnson-Cook, Zerilli-Armstrong or Simo. We discuss the different forms of the laws and their taking into account of strain rates, speed and temperature. The iterative procedure for the identification uses an experimental set of parameters (from a Taylor impact test) which are compared with an other set of numerical values. The method to converge to the optimal solution uses a combined Monte-Carlo and Levenberg-Marquardt algorithm. The study ends with a comparative discussion about the laws and the problems encountered during their identification for two materials: a 42CD4 steel, and an aluminum 2017T3.

  3. High Strain Rate Deformation Modeling of a Polymer Matrix Composite. Part 1; Matrix Constitutive Equations

    Science.gov (United States)

    Goldberg, Robert K.; Stouffer, Donald C.

    1998-01-01

    Recently applications have exposed polymer matrix composite materials to very high strain rate loading conditions, requiring an ability to understand and predict the material behavior under these extreme conditions. In this first paper of a two part report, background information is presented, along with the constitutive equations which will be used to model the rate dependent nonlinear deformation response of the polymer matrix. Strain rate dependent inelastic constitutive models which were originally developed to model the viscoplastic deformation of metals have been adapted to model the nonlinear viscoelastic deformation of polymers. The modified equations were correlated by analyzing the tensile/ compressive response of both 977-2 toughened epoxy matrix and PEEK thermoplastic matrix over a variety of strain rates. For the cases examined, the modified constitutive equations appear to do an adequate job of modeling the polymer deformation response. A second follow-up paper will describe the implementation of the polymer deformation model into a composite micromechanical model, to allow for the modeling of the nonlinear, rate dependent deformation response of polymer matrix composites.

  4. IFN-α Is Constitutively Expressed in the Human Thymus, but Not in Peripheral Lymphoid Organs

    NARCIS (Netherlands)

    Colantonio, A.D.; Epeldegui, M.; Jesiak, M.; Jachimowski, L.; Blom, B.; Uittenbogaart, C.H.

    2011-01-01

    Type I interferons have been typically studied for their effects in the context of bacterial or viral infections. However in this report, we provide evidence that Interferon-alpha (IFN-α) expressing cells are present in the thymus in the absence of infection. We show that pDC express the highest

  5. IFN-α Is Constitutively Expressed in the Human Thymus, but Not in Peripheral Lymphoid Organs

    NARCIS (Netherlands)

    Colantonio, A.D.; Epeldegui, M.; Jesiak, M.; Jachimowski, L.; Blom, B.; Uittenbogaart, C.H.

    2011-01-01

    Type I interferons have been typically studied for their effects in the context of bacterial or viral infections. However in this report, we provide evidence that Interferon-alpha (IFN-α) expressing cells are present in the thymus in the absence of infection. We show that pDC express the highest lev

  6. Prediction of multiaxial mechanical behavior for conventional and highly crosslinked UHMWPE using a hybrid constitutive model.

    Science.gov (United States)

    Bergström, J S; Rimnac, C M; Kurtz, S M

    2003-04-01

    The development of theoretical failure, fatigue, and wear models for ultra-high molecular weight polyethylene (UHMWPE) used in joint replacements has been hindered by the lack of a validated constitutive model that can accurately predict large deformation mechanical behavior under clinically relevant, multiaxial loading conditions. Recently, a new Hybrid constitutive model for unirradiated UHMWPE was developed Bergström et al., (Biomaterials 23 (2002) 2329) based on a physics-motivated framework which incorporates the governing micro-mechanisms of polymers into an effective and accurate continuum representation. The goal of the present study was to compare the predictive capability of the new Hybrid model with the J(2)-plasticity model for four conventional and highly crosslinked UHMWPE materials during multiaxial loading. After calibration under uniaxial loading, the predictive capabilities of the J(2)-plasticity and Hybrid model were tested by comparing the load-displacement curves from experimental multiaxial (small punch) tests with simulated load-displacement curves calculated using a finite element model of the experimental apparatus. The quality of the model predictions was quantified using the coefficient of determination (r(2)). The results of the study demonstrate that the Hybrid model outperforms the J(2)-plasticity model both for combined uniaxial tension and compression predictions and for simulating multiaxial large deformation mechanical behavior produced by the small punch test. The results further suggest that the parameters of the HM may be generalizable for a wide range of conventional, highly crosslinked, and thermally treated UHMWPE materials, based on the characterization of four material properties related to the elastic modulus, yield stress, rate of strain hardening, and locking stretch of the polymer chains. Most importantly, from a practical perspective, these four key material properties for the Hybrid constitutive model can be measured

  7. Inhibition of human UGT2B7 gene expression in transgenic mice by the constitutive androstane receptor.

    Science.gov (United States)

    Yueh, M F; Mellon, P L; Tukey, R H

    2011-06-01

    The xenobiotic receptors, constitutive androstane receptor (CAR), and pregnane X receptor (PXR) regulate and alter the metabolism of xenobiotic substrates. Among the 19 functional UDP-glucuronosyltransferases (UGTs) in humans, UGT2B7 is involved in the metabolism of many structurally diverse xenobiotics and plays an important role in the clearance and detoxification of many therapeutic drugs. To examine whether this gene is regulated by CAR and PXR in vivo, transgenic mice expressing the entire UGT2B7 gene (TgUGT2B7) were created. Gene expression profiles revealed that UGT2B7 is differentially expressed in liver, kidney, adipocytes, brain, and estrogen-sensitive tissues, such as ovary and uterus. Liver UGT2B7 expression levels were decreased when TgUGT2B7 mice were treated with the CAR ligand 1,4-b-s-[2-(3,5,-dichloropyridyloxy)] (TCPOBOP) but not the PXR ligand pregnenolone 16α-carbonitrile. Although TCPOBOP decreased the levels of UGT2B7 mRNA in TgUGT2B7 mice, it had no affect on Tg(UGT2B7)Car(-/-) mice, adding support for a CAR-dependent mechanism contributing toward UGT2B7 gene suppression. Expression of promoter constructs in HepG2 cells showed the CAR-dependent inhibition was linked to hepatocyte nuclear factor-4α (HNF4α)-mediated transactivation of the UGT2B7 promoter. The inhibitory effect of CAR on UGT2B7 gene expression was validated in chromatin immunoprecipitation assays in which TCPOBOP treatment blocked HNF4α binding to the UGT2B7 promoter. These results suggest that HNF4α plays an important role in the constitutive expression of hepatic UGT2B7, and CAR acts as a negative regulator by interfering with HNF4α binding activity.

  8. Constitutive expression of human coagulating factor IX in HeLa cells by homologous recombination of the promoter

    Institute of Scientific and Technical Information of China (English)

    2001-01-01

    Constitutive expression of hFIX protein in nonhepatocytes wasstudied. The gene targeting vector was constructed and transferred into HeLa cells. With the detection system of PCR, we demonstrated that the endogenous hFIX promoter was replaced with an hCMV promoter when targeted insertion of the constructor was directed by the sequence homology. The expression of hFIX in the modified HeLa cells, 11.2 ng/106 cell/24 h, strongly suggested that hFIX gene could be activated by a powerful promoter in nonhepatocytes. The results would make it possible to examine the feasibility of re-regulate gene expression by promoter replacement.

  9. Expression of a constitutively active prolactin receptor causes histone trimethylation of the p53 gene in breast cancer

    Institute of Scientific and Technical Information of China (English)

    Tan Dunyong; Tang Peizhi; Huang Jianjun; Zhang Jie; Zhou Weihua; Ameae M.Walker

    2014-01-01

    Background Prolactin (PRL) is a pituitary polypeptide hormone characterized by multiple biological actions including stimulation of growth in the prostate and formation of secretory alveoli and stimulation of milk protein gene expression in the mammary gland.PRL exerts its effect by dimerizing its receptor (PRLR) on the plasma membrane and regulating gene expression through the JAK-Stat signal pathway.We have previously described a natural variant of the PRLR in which the S2 subdomain of the extracellular domain is missing (Delta S2).Delta S2 PRLRs are dimerized in the absence of PRL and have constitutive activity in the promotion of breast cancer cell growth.Enhancer of zeste homolog 2 (EZH2),as one of the histone-modifying enzymes,is a key factor regulating gene expression by epigenetic modification.We hypothesized that these constitutive activated Delta S2 PRLRs played a pathogenic role in breast cancer in part through alterations in the expression of EZH2 and the trimethylation of histone 3 on lysine 27 (H3K27Me3).Methods In order to verify the clinical significance and to establish the link between Delta S2 PRLR expression and epigenetic change,EZH2,H3K27Me3,and Delta S2 PRLR were detected in both normal and cancerous human breast tissues.Also,overexpression of Delta S2 PRLR in breast epithelial cells was achieved by infection with adenovirus carrying the cDNA.Western blotting and chromatin immunoprecipitation (ChIP assay) and acid histone extraction were applied to detect the expression of EZH2 and the trimethylation of histone 3,respectively.Results In breast tissue,higher EZH2 expression and higher H3K27Me3 were found associated with higher Delta S2 expression in breast cancer samples.In breast epithelial cells,overexpression of Delta S2 PRLR increased EZH2 methyltransferase mRNA and protein,induced EZH2 methyltransferase recruitment to chromatin,increased the trimethylation of H3K27Me3,and decreased the expression of p53 gene.Conclusions Delta S2 PRLR

  10. Constitutive expression of a costimulatory ligand on antigen-presenting cells in the nervous system drives demyelinating disease

    DEFF Research Database (Denmark)

    Zehntner, Simone P; Brisebois, Marcel; Tran, Elise

    2003-01-01

    that transgenic mice constitutively expressing the costimulatory ligand B7.2/CD86 on microglia in the central nervous system (CNS) and on related cells in the proximal peripheral nervous tissue spontaneously develop autoimmune demyelinating disease. Disease-affected nervous tissue in transgenic mice showed...... recipients but not into non-transgenic recipients. These data provide evidence that B7/CD28 interactions within the nervous tissue are critical determinants of disease development. Our findings have important implications for understanding the etiology of nervous system autoimmune diseases such as multiple...

  11. Establishment of Sf9 transformants constitutively expressing PBAN receptor variants: application to functional evaluation

    Science.gov (United States)

    To facilitate further evaluation of pheromone biosynthesis activating neuropeptide receptor (PBANR) functionality and regulation, we generated cultured insect cell lines stably expressing a number of fluorescent Bombyx mori PBANR (BommoPBANR) and Pseudaletia separata PBANR (PsesePBANR) variants incl...

  12. Constitutive expression of inducible nitric oxide synthase in the normal human colonic epithelium

    DEFF Research Database (Denmark)

    Perner, A; Andresen, L; Normark, M

    2002-01-01

    Inducible nitric oxide synthase (iNOS) in the human colon is considered expressed only in inflammatory states such as ulcerative or collagenous colitis. As subtle iNOS labelling was previously observed in some colonic mucosal biopsies from a heterogeneous group of controls with non-inflamed bowel...

  13. Constitutive expression of inducible nitric oxide synthase in the normal human colonic epithelium

    DEFF Research Database (Denmark)

    Perner, A; Andresen, Lars; Normark, M;

    2002-01-01

    Inducible nitric oxide synthase (iNOS) in the human colon is considered expressed only in inflammatory states such as ulcerative or collagenous colitis. As subtle iNOS labelling was previously observed in some colonic mucosal biopsies from a heterogeneous group of controls with non-inflamed bowel...

  14. Suppression of granzyme B activity and caspase-3 activation in leukaemia cells constitutively expressing the protease inhibitor 9.

    Science.gov (United States)

    Fritsch, Kristina; Finke, Jürgen; Grüllich, Carsten

    2013-12-01

    Immune surveillance against malignant cells is mediated by cytotoxic T-lymphocytes and NK-cells (CTL/NK) that induce apoptosis through the granzyme-B-dependent pathway. The serine protease inhibitor serpinB9/protease inhibitor-9 (PI-9) is a known inhibitor of granzyme B. Ectopic expression of PI-9 in tumour cells has been reported. However, the impact of PI-9 on granzyme-B-induced apoptosis in tumour cells remains unclear. The aim of this study was to investigate the influence of constitutive PI-9 expression in leukaemia cell lines on the activity of granzyme B and apoptosis induction. PI-9 negative (lymphoblastic Jurkat cells; myeloblastic U937 cells) and PI-9-expressing cell lines (myeloblastic K562 cells, EBV-transformed LCL-1 and LCL-2 B-cells, lymphoblastic Daudi cells, AML-R cells f leukaemia and the U937 subclone U937PI-9(+)). For accurate granzyme B activity determination a quantitative substrate (Ac-IEPD-pNA) cleavage assay was established and caspase-3 activation measured for apoptosis assessment. Cells were treated with a cytotoxic granule isolate that has previously been shown to induce apoptosis through granzyme B signalling. We found a robust correlation between constitutive PI-9 expression levels and the suppression of granzyme B activity. Further, inhibition of granzyme B translated into reduced caspase-3 activation. We conclude, suppression of granzyme B initiated apoptosis in PI-9-expressing cells could contribute to immune evasion and the measurement of granzyme B activity with our assay might be a useful predictive marker in immune-therapeutic approaches against cancer.

  15. Selective lignin downregulation leads to constitutive defense response expression in alfalfa (Medicago sativa L.).

    Science.gov (United States)

    Gallego-Giraldo, Lina; Jikumaru, Yusuke; Kamiya, Yuji; Tang, Yuhong; Dixon, Richard A

    2011-05-01

    • Downregulation of hydroxycinnamoyl CoA: shikimate hydroxycinnamoyl transferase (HCT) in alfalfa (Medicago sativa) reduces lignin levels and improves forage quality and saccharification efficiency for bioethanol production. However, the plants have reduced stature. It was previously reported that HCT-down-regulated Arabidopsis have impaired auxin transport, but this has recently been disproved. • To address the basis for the phenotypes of lignin-modified alfalfa, we measured auxin transport, profiled a range of metabolites including flavonoids and hormones, and performed in depth transcriptome analyses. • Auxin transport is unaffected in HCT antisense alfalfa despite increased flavonoid biosynthesis. The plants show increased cytokinin and reduced auxin levels, and gibberellin levels and sensitivity are both reduced. Levels of salicylic, jasmonic and abscisic acids are elevated, associated with massive upregulation of pathogenesis and abiotic stress-related genes and enhanced tolerance to fungal infection and drought. • We suggest that HCT downregulated alfalfa plants exhibit constitutive activation of defense responses, triggered by release of bioactive cell wall fragments and production of hydrogen peroxide as a result of impaired secondary cell wall integrity. © 2011 The Authors. New Phytologist © 2011 New Phytologist Trust.

  16. A Damaged Constitutive Model for Rock under Dynamic and High Stress State

    Directory of Open Access Journals (Sweden)

    Yan-Long Li

    2017-01-01

    Full Text Available The main research work of this paper focuses on the theoretical prediction of the constitutive relationship for rock, concrete, and other quasi-brittle materials under dynamic and complex stress state and the influence of dynamic mechanical behavior of rock on practical engineering problems was studied. A damaged elastoplastic model (DEPM is established for the investigation and prediction of static or dynamic mechanical behavior of rock material. The mechanical behavior (brittleness or plasticity and dynamic response (due to underground impact pressure and high-velocity impact of projectile of rock under high in situ stress were investigated via the DEPM combined with the explicit finite element method. This paper suggests the influence of the brittle or plastic mechanical behavior of rock material on deep underground rock engineering.

  17. Parameter correlation of high-temperature creep constitutive equation for RPV metallic materials

    Energy Technology Data Exchange (ETDEWEB)

    Xie, Lin-Jun, E-mail: linjunx@zjut.edu.cn; Ren, Xin; Shen, Ming-Xue; Tu, Li-Qun

    2015-10-15

    Constant-temperature and constant-load creep tests of SA-508 stainless steel were performed at six temperatures, and the creep behavior and properties of this material were determined. Constitutive models were established based on an isothermal creep method to describe the high-temperature creep behavior of SA-508. Material parameter k, stress exponent n{sub σ}, and temperature exponent n{sub t} of the established constitutive models were determined through experimental data via numerical optimization techniques. The relationship of k, n{sub σ}, and n{sub t} was evaluated, and a new coefficient model of k–T, n{sub σ}–T, n{sub t}–T, and n{sub t}–n{sub σ} was formulated through the parameters of the isothermal creep equation. Moreover, the isothermal creep equation for this material at every temperature point from 450 °C to 1000 °C was obtained from the models. This method can serve as a reference for isothermal creep analysis and provide a way for the safety assessment of components of reactor pressure vessels.

  18. High constitutive signaling of the ghrelin receptor--identification of a potent inverse agonist

    DEFF Research Database (Denmark)

    Holst, Birgitte; Cygankiewicz, Adam; Jensen, Tine Halkjaer;

    2003-01-01

    Ghrelin is a GH-releasing peptide that also has an important role as an orexigenic hormone-stimulating food intake. By measuring inositol phosphate turnover or by using a reporter assay for transcriptional activity controlled by cAMP-responsive elements, the ghrelin receptor showed strong, ligand...... and appetite control. It is suggested that inverse agonists for the ghrelin receptor could be particularly interesting for the treatment of obesity.......Ghrelin is a GH-releasing peptide that also has an important role as an orexigenic hormone-stimulating food intake. By measuring inositol phosphate turnover or by using a reporter assay for transcriptional activity controlled by cAMP-responsive elements, the ghrelin receptor showed strong, ligand...... found to be a high potency (EC50 = 5.2 nm) full inverse agonist as it decreased the constitutive signaling of the ghrelin receptor down to that observed in untransfected cells. The homologous motilin receptor functioned as a negative control as it did not display any sign of constitutive activity...

  19. A dedicated promoter drives constitutive expression of the cell-autonomous immune resistance GTPase, Irga6 (IIGP1 in mouse liver.

    Directory of Open Access Journals (Sweden)

    Jia Zeng

    Full Text Available BACKGROUND: In general, immune effector molecules are induced by infection. METHODOLOGY AND PRINCIPAL FINDINGS: However, strong constitutive expression of the cell-autonomous resistance GTPase, Irga6 (IIGP1, was found in mouse liver, contrasting with previous evidence that expression of this protein is exclusively dependent on induction by IFNgamma. Constitutive and IFNgamma-inducible expression of Irga6 in the liver were shown to be dependent on transcription initiated from two independent untranslated 5' exons, which splice alternatively into the long exon encoding the full-length protein sequence. Irga6 is expressed constitutively in freshly isolated hepatocytes and is competent in these cells to accumulate on the parasitophorous vacuole membrane of infecting Toxoplasma gondii tachyzoites. CONCLUSIONS AND SIGNIFICANCE: The role of constitutive hepatocyte expression of Irga6 in resistance to parasites invading from the gut via the hepatic portal system is discussed.

  20. Constitutive expression of pathogenesis-related proteins and antioxydant enzyme activities triggers maize resistance towards Fusarium verticillioides.

    Science.gov (United States)

    Maschietto, Valentina; Lanubile, Alessandra; Leonardis, Silvana De; Marocco, Adriano; Paciolla, Costantino

    2016-08-01

    Fusarium verticillioides is a fungal pathogen of maize that causes ear rot and contaminates the grains with fumonisin mycotoxins. Breeding for resistance to Fusarium emerged as the most economic and environmentally safe strategy; therefore the discovery of resistant sources and effective molecular markers are a priority. Ears of resistant (CO441 and CO433) and susceptible (CO354 and CO389) maize lines were inoculated with F. verticillioides and the expression of pathogenesis-related (PR) genes (PR1, PR5, PRm3, PRm6) and genes that protect from oxidative stress (peroxidase, catalase, superoxide dismutase and ascorbate peroxidase) were evaluated in the kernels at 72h post inoculation. In addition, the oxidation level and the enzymatic activity of ascorbate-glutathione cycle, catalase, superoxide dismutase and cytosolic and wall peroxidases were investigated. The uninoculated kernels of the resistant lines showed higher gene expression and enzymatic activities, highlighting the key role of constitutive resistance in limiting pathogen attack. In contrast, the susceptible lines activated defensive genes only after pathogen inoculation, resulting in increased levels of H2O2 and lipid peroxidation, as well as lower enzymatic activities. The constitutive defenses observed in this study from seed could be profitably exploited to develop markers to speed up conventional breeding programs in the selection of resistant genotypes. Copyright © 2016 Elsevier GmbH. All rights reserved.

  1. Retinoic acid-inducible gene-I is constitutively expressed and involved in IFN-gamma-stimulated CXCL9-11 production in intestinal epithelial cells.

    Science.gov (United States)

    Kawaguchi, Shogo; Ishiguro, Yoh; Imaizumi, Tadaatsu; Mori, Fumiaki; Matsumiya, Tomoh; Yoshida, Hidemi; Ota, Ken; Sakuraba, Hirotake; Yamagata, Kazufumi; Sato, Yuki; Tanji, Kunikazu; Haga, Toshihiro; Wakabayashi, Koichi; Fukuda, Shinsaku; Satoh, Kei

    2009-03-24

    Retinoic acid-inducible gene-I (RIG-I) is a member of the DExH/D family proteins, and plays an important role in antiviral response via interferon-stimulated genes (ISGs) and type 1 IFN. In this study, the roles of RIG-I in the epithelial cells in the cross-talk between type 2 IFN and inducible chemokines production are high-lighted. The results showed that RIG-I was constitutively expressed in normal surface epithelia lining the colonic mucosa. RIG-I was constitutively expressed in the epithelial cell lines HT-29, and IFN-gamma and TNF-alpha enhanced the RIG-I expression in a dose-dependent manner. IFN-gamma was shown to stimulate CXCL9-11 production, and RNA interference against RIG-I resulted in significant decrease of IFN-gamma-induced CXCL9-11 productions. These results suggest that RIG-I play an important role in the cross-talk between inflammatory cytokines and immune cell trafficking. In conclusion, RIG-I might regulate the gut barrier function in homeostatic and inflammatory conditions.

  2. Expression of a constitutively active nitrate reductase variant in tobacco reduces tobacco-specific nitrosamine accumulation in cured leaves and cigarette smoke.

    Science.gov (United States)

    Lu, Jianli; Zhang, Leichen; Lewis, Ramsey S; Bovet, Lucien; Goepfert, Simon; Jack, Anne M; Crutchfield, James D; Ji, Huihua; Dewey, Ralph E

    2016-07-01

    Burley tobaccos (Nicotiana tabacum) display a nitrogen-use-deficiency phenotype that is associated with the accumulation of high levels of nitrate within the leaf, a trait correlated with production of a class of compounds referred to as tobacco-specific nitrosamines (TSNAs). Two TSNA species, 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanone (NNK) and N-nitrosonornicotine (NNN), have been shown to be strong carcinogens in numerous animal studies. We investigated the potential of molecular genetic strategies to lower nitrate levels in burley tobaccos by overexpressing genes encoding key enzymes of the nitrogen-assimilation pathway. Of the various constructs tested, only the expression of a constitutively active nitrate reductase (NR) dramatically decreased free nitrate levels in the leaves. Field-grown tobacco plants expressing this NR variant exhibited greatly reduced levels of TSNAs in both cured leaves and mainstream smoke of cigarettes made from these materials. Decreasing leaf nitrate levels via expression of a constitutively active NR enzyme represents an exceptionally promising means for reducing the production of NNN and NNK, two of the most well-documented animal carcinogens found in tobacco products. © 2016 The Authors. Plant Biotechnology Journal published by Society for Experimental Biology and The Association of Applied Biologists and John Wiley & Sons Ltd.

  3. Waking up Streptomyces secondary metabolism by constitutive expression of activators or genetic disruption of repressors.

    Science.gov (United States)

    Aigle, Bertrand; Corre, Christophe

    2012-01-01

    Streptomycete bacteria are renowned as a prolific source of natural products with diverse biological activities. Production of these metabolites is often subject to transcriptional regulation: the biosynthetic genes remain silent until the required environmental and/or physiological signals occur. Consequently, in the laboratory environment, many gene clusters that direct the biosynthesis of natural products with clinical potential are not expressed or at very low level preventing the production/detection of the associated metabolite. Genetic engineering of streptomycetes can unleash the production of many new natural products. This chapter describes the overexpression of pathway-specific activators, the genetic disruption of pathway-specific repressors, and the main strategy used to identify and characterize new natural products from these engineered Streptomyces strains.

  4. Plenary perspective: the complexity of constitutive and inducible gene expression in mononuclear phagocytes.

    Science.gov (United States)

    Hume, David A

    2012-09-01

    Monocytes and macrophages differentiate from progenitor cells under the influence of colony-stimulating factors. Genome-scale data have enabled the identification of the set of genes that distinguishes macrophages from other cell types and the ways in which thousands of genes are regulated in response to pathogen challenge. Although there has been a focus on a small subset of lineage-enriched transcription factors, such as PU.1, more than one-half of the transcription factors in the genome can be expressed in macrophage lineage cells under some state of activation, and they interact in a complex network. The network architecture is conserved across species, but many of the target genes evolve rapidly and differ between mouse and human. The data and publication deluge related to macrophage biology require the development of new analytical tools and ways of presenting information in an accessible form.

  5. TRP 9904 - Constitutive Behavior of High Strength Multiphase Sheel Steel Under High Strain Rate Deformation

    Energy Technology Data Exchange (ETDEWEB)

    David Matlock; John Speer

    2005-03-31

    The focus of the research project was to systematically assess the strain rate dependence of strengthening mechanisms in new advanced high strength sheet steels. Data were obtained on specially designed and produced Duel Phase and TRIP steels and compared to the properties of automotive steels currently in use.

  6. Constitutive expression of a plant ferredoxin-like protein (pflp) enhances capacity of photosynthetic carbon assimilation in rice (Oryza sativa).

    Science.gov (United States)

    Chang, Hsiang; Huang, Hsiang-En; Cheng, Chin-Fu; Ho, Mei-Hsuan; Ger, Mang-Jye

    2017-04-01

    The plant ferredoxin-like protein (PFLP) gene, cloned from sweet peppers predicted as an electron carrier in photosynthesis, shows high homology to the Fd-I sequence of Arabidopsis thaliana, Lycopersicon esculentum, Oryza sativa and Spinacia oleracea. Most of pflp related studies focused on anti-pathogenic effects, while less understanding for the effects in photosynthesis with physiological aspects, such as photosynthesis rate, and levels of carbohydrate metabolites. This project focuses on the effects of pflp overexpression on photosynthesis by physiological evaluations of carbon assimilation with significant higher levels of carbohydrates with higher photosynthesis efficiency. In this report, two independent transgenic lines of rice plants (designated as pflp-1 and pflp-2) were generated from non-transgenic TNG67 rice plant (WT). Both transgenic pflp rice plants exhibited enhanced photosynthesis efficiency, and gas exchange rates of photosynthesis were 1.3- and 1.2-fold higher for pflp-1 and pflp-2 than WT respectively. Significantly higher electron transport rates of pflp rice plants were observed. Moreover, photosynthetic products, such as fructose, glucose, sucrose and starch contents of pflp transgenic lines were increased accordingly. Molecular evidences of carbohydrate metabolism related genes activities (osHXK5, osHXK6, osAGPL3, osAGPS2α, osSPS, ospFBPase, oscFBPase, and osSBPase) in transgenic lines were higher than those of WT. For performance of crop production, 1000-grain weight for pflp-1 and pflp-2 rice plants were 52.9 and 41.1 g that were both significantly higher than 31.6 g for WT, and panicles weights were 1.4- and 1.2-fold higher than WT. Panicle number, tiller number per plants for pflp rice plants were all significantly higher compared with those of WT where there was no significant difference observed between two pflp rice plants. Taken altogether; this study demonstrated that constitutive pflp expression can improve rice production by

  7. Differential requirements of two recA mutants for constitutive SOS expression in Escherichia coli K-12.

    Directory of Open Access Journals (Sweden)

    Jarukit Edward Long

    Full Text Available BACKGROUND: Repairing DNA damage begins with its detection and is often followed by elicitation of a cellular response. In E. coli, RecA polymerizes on ssDNA produced after DNA damage and induces the SOS Response. The RecA-DNA filament is an allosteric effector of LexA auto-proteolysis. LexA is the repressor of the SOS Response. Not all RecA-DNA filaments, however, lead to an SOS Response. Certain recA mutants express the SOS Response (recA(C in the absence of external DNA damage in log phase cells. METHODOLOGY/PRINCIPAL FINDINGS: Genetic analysis of two recA(C mutants was used to determine the mechanism of constitutive SOS (SOS(C expression in a population of log phase cells using fluorescence of single cells carrying an SOS reporter system (sulAp-gfp. SOS(C expression in recA4142 mutants was dependent on its initial level of transcription, recBCD, recFOR, recX, dinI, xthA and the type of medium in which the cells were grown. SOS(C expression in recA730 mutants was affected by none of the mutations or conditions tested above. CONCLUSIONS/SIGNIFICANCE: It is concluded that not all recA(C alleles cause SOS(C expression by the same mechanism. It is hypothesized that RecA4142 is loaded on to a double-strand end of DNA and that the RecA filament is stabilized by the presence of DinI and destabilized by RecX. RecFOR regulate the activity of RecX to destabilize the RecA filament. RecA730 causes SOS(C expression by binding to ssDNA in a mechanism yet to be determined.

  8. Differential requirements of two recA mutants for constitutive SOS expression in Escherichia coli K-12.

    Science.gov (United States)

    Long, Jarukit Edward; Renzette, Nicholas; Centore, Richard C; Sandler, Steven J

    2008-01-01

    Repairing DNA damage begins with its detection and is often followed by elicitation of a cellular response. In E. coli, RecA polymerizes on ssDNA produced after DNA damage and induces the SOS Response. The RecA-DNA filament is an allosteric effector of LexA auto-proteolysis. LexA is the repressor of the SOS Response. Not all RecA-DNA filaments, however, lead to an SOS Response. Certain recA mutants express the SOS Response (recA(C)) in the absence of external DNA damage in log phase cells. Genetic analysis of two recA(C) mutants was used to determine the mechanism of constitutive SOS (SOS(C)) expression in a population of log phase cells using fluorescence of single cells carrying an SOS reporter system (sulAp-gfp). SOS(C) expression in recA4142 mutants was dependent on its initial level of transcription, recBCD, recFOR, recX, dinI, xthA and the type of medium in which the cells were grown. SOS(C) expression in recA730 mutants was affected by none of the mutations or conditions tested above. It is concluded that not all recA(C) alleles cause SOS(C) expression by the same mechanism. It is hypothesized that RecA4142 is loaded on to a double-strand end of DNA and that the RecA filament is stabilized by the presence of DinI and destabilized by RecX. RecFOR regulate the activity of RecX to destabilize the RecA filament. RecA730 causes SOS(C) expression by binding to ssDNA in a mechanism yet to be determined.

  9. Particle characteristics and rheological constitutive relations of high concentration red mud

    Institute of Scientific and Technical Information of China (English)

    WANG Xing; QU Yuan-yuan; HU Wei-wei; CHEN Jie; ZHAO Xue-yi; WU Miao

    2008-01-01

    Red mud has relatively small solid particles (d50=13.02 μm) and will flow in paste form under high pressure during pipeline transport. Red mud belongs to a two-phase flow of materials with high viscosity and a high concentration of non-sedimentation, homogeneous solid-liquids. It is difficult to test its rheological properties under atmospheric pressure. Measurements such as rotational viscometry can not reflect the real state of the material when it is flowing in a pipe. Tested rheological parameters are somewhat higher than the actual values. In our investigation, grain shape, distinctive modality and grain size distribution of red mud were tested. Based on the principle of tube measurement, rheological experiments on red mud at different concentrations were carried out by using our independently developed tube-type pressure rheology test facility, and obtained constitutive equations. We conclude that red mud behaves as non-Newtonian pseudo-plastic fluid in pipe flows. Its consistency and power-law indices vary considerably with different concentrations.

  10. High-Resolution Multispectral Optoacoustic Tomography of the Vascularization and Constitutive Hypoxemia of Cancerous Tumors

    Directory of Open Access Journals (Sweden)

    Andrei Chekkoury

    2016-08-01

    Full Text Available Diversity of the design and alignment of illumination and ultrasonic transducers empower the fine scalability and versatility of optoacoustic imaging. In this study, we implement an innovative high-resolution optoacoustic mesoscopy for imaging the vasculature and tissue oxygenation within subcutaneous and orthotopic cancerous implants of mice in vivo through acquisition of tomographic projections over 180° at a central frequency of 24 MHz. High-resolution volumetric imaging was combined with multispectral functional measurements to resolve the exquisite inner structure and vascularization of the entire tumor mass using endogenous and exogenous optoacoustic contrast. Evidence is presented for constitutive hypoxemia within the carcinogenic tissue through analysis of the hemoglobin absorption spectra and distribution. Morphometric readouts obtained with optoacoustic mesoscopy have been verified with high-resolution ultramicroscopic studies. The findings described herein greatly extend the applications of optoacoustic mesoscopy toward structural and multispectral functional measurements of the vascularization and hemodynamics within solid tumors in vivo and are of major relevance to basic and preclinical oncological studies in small animal models.

  11. Ectopic Expression of CsCTR1, a Cucumber CTR-Like Gene, Attenuates Constitutive Ethylene Signaling in an Arabidopsis ctr1-1 Mutant and Expression Pattern Analysis of CsCTR1 in Cucumber (Cucumis sativus

    Directory of Open Access Journals (Sweden)

    Beibei Bie

    2014-09-01

    Full Text Available The gaseous plant hormone ethylene regulates many aspects of plant growth, development and responses to the environment. Constitutive triple response 1 (CTR1 is a central regulator involved in the ethylene signal transduction pathway. To obtain a better understanding of this particular pathway in cucumber, the cDNA-encoding CTR1 (designated CsCTR1 was isolated from cucumber. A sequence alignment and phylogenetic analyses revealed that CsCTR1 has a high degree of homology with other plant CTR1 proteins. The ectopic expression of CsCTR1 in the Arabidopsis ctr1-1 mutant attenuates constitutive ethylene signaling of this mutant, suggesting that CsCTR1 indeed performs its function as negative regulator of the ethylene signaling pathway. CsCTR1 is constitutively expressed in all of the examined cucumber organs, including roots, stems, leaves, shoot apices, mature male and female flowers, as well as young fruits. CsCTR1 expression gradually declined during male flower development and increased during female flower development. Additionally, our results indicate that CsCTR1 can be induced in the roots, leaves and shoot apices by external ethylene. In conclusion, this study provides a basis for further studies on the role of CTR1 in the biological processes of cucumber and on the molecular mechanism of the cucumber ethylene signaling pathway.

  12. A constitutive model for particulate-reinforced titanium matrix composites subjected to high strain rates and high temperatures

    Directory of Open Access Journals (Sweden)

    Song Wei-Dong

    2013-01-01

    Full Text Available Quasi-static and dynamic tension tests were conducted to study the mechanical properties of particulate-reinforced titanium matrix composites at strain rates ranging from 0.0001/s to 1000/s and at temperatures ranging from 20 °C to 650 °C Based on the experimental results, a constitutive model, which considers the effects of strain rate and temperature on hot deformation behavior, was proposed for particulate-reinforced titanium matrix composites subjected to high strain rates and high temperatures by using Zener-Hollomon equations including Arrhenius terms. All the material constants used in the model were identified by fitting Zener-Hollomon equations against the experimental results. By comparison of theoretical predictions presented by the model with experimental results, a good agreement was achieved, which indicates that this constitutive model can give an accurate and precise estimate for high temperature flow stress for the studied titanium matrix composites and can be used for numerical simulations of hot deformation behavior of the composites.

  13. Constitutively expressed Protocadherin-α regulates the coalescence and elimination of homotypic olfactory axons through its cytoplasmic region

    Directory of Open Access Journals (Sweden)

    Sonoko eHasegawa

    2012-10-01

    Full Text Available Olfactory sensory neuron (OSN axons coalesce into specific glomeruli in the olfactory bulb (OB according to their odorant receptor (OR expression. Several guidance molecules enhance the coalescence of homotypic OSN projections, in an OR-specific- and neural-activity-dependent manner. However, the mechanism by which homotypic OSN axons are organized into glomeruli is unsolved. We previously reported that the clustered protocadherin-α (Pcdh-α family of diverse cadherin-related molecules plays roles in the coalescence and elimination of homotypic OSN axons throughout development. Here we showed that the elimination of small ectopic homotypic glomeruli required the constitutive expression of a Pcdh-α isoform and Pcdh-α’s cytoplasmic region, but not OR specificity or neural activity. These results suggest that Pcdh-α proteins provide a cytoplasmic signal to regulate repulsive activity for homotypic OSN axons independently of OR expression and neural activity. The counterbalancing effect of Pcdh-α proteins for the axonal coalescence mechanisms mediated by other olfactory guidance molecules indicate a possible mechanism for the organization of homotypic OSN axons into glomeruli during development.

  14. Construction and characterization of a hrpG mutant rendering constitutive expression of hrp genes in Xanthomonas campestris pv. campestris

    Institute of Scientific and Technical Information of China (English)

    JIANG Bole; XU Rongqi; LI Xianzhen; WEI Hongyu; BAI Faan; HU Xi; HE Yongqiang; TANG Jiliang

    2006-01-01

    The expression of hrp genes in Xanthomonas is regulated by hrpG, an OmpR-type transcriptional activator, and can be induced by artificial mimic medium XVM2, but repressed in rich medium NYG. In this study, a site-specific mutation of hrpG in Xanthomonas campestris pv. campestris (Xcc) was generated by PCR-directed point mutagenesis and was characterized by its phenotype and transcriptional expression analyses. The results indicated that the E45K mutated hrpG rendered all the hrp genes expressing constitutively in the suppression medium NYG. Plant test showed that mutated hrpG enhanced the timing and intensity of hypersensitive reaction (HR) on the nonhost plant pepper, but did not affect the growth of Xcc in both culture medium and plants. The E45K mutation did not cause significant changes of production of exopolysaccharide, extracellular cellulase and amylase, but it inhibited the secretion of extracellular protease. This mutant 8004 * is more suitable than the wild type for identifying hrp-dependent effectors in vitro and studying the transcriptional regulation genome-wide.

  15. De novo synthesis, constitutive expression of Aspergillus sulphureus beta-xylanase gene in Pichia pastoris and partial enzymic characterization.

    Science.gov (United States)

    Cao, Yunhe; Qiao, Jiayun; Li, Yihang; Lu, Wenqing

    2007-09-01

    The endo-beta-1, 4-xylanase gene xynA from Aspergillus sulphureus, encoded a lack-of-signal peptide protein of 184 amino acids, was de novo synthesized by splicing overlap extension polymerase chain reaction according to Pichia pastoris protein's codon bias. The synthetic DNA, composed of 572 nucleotides, was ligated into the downstream sequence of an alpha-mating factor in a constitutive expression vector pGAPzalphaA and electrotransformed into the P. pastoris X-33 strain. The transformed yeast screened by Zeocin was able to constitutively secrete the xylanase in yeast-peptone-dextrose liquid medium. The heterogenous DNA was stabilized in the strain by 20-times passage culture. The recombinant enzyme was expressed with a yield of 120 units/mL under the flask culture at 28 degrees C for 3 days. The enzyme showed optimal activity at 50 degrees C and pH 2.4-3.4. Residual activity of the raw recombinant xylanase was not less than 70% when fermentation broth was directly heated at 80 degrees C for 30 min. However, the dialyzed xylanase supernatant completely lost the catalytic activity after being heated at 60 degrees C for 30 min. The recombinant xylanase showed no obvious activity alteration by being pretreated with Na(2)HPO(4)-citric acid buffer of pH 2.4 for 2 h. The xylanase also showed resistance to certain metal ions (Na(+), Mg(2+), Ca(2+), K(+), Ba(2+), Zn(2+), Fe(2+), and Mn(2+)) and EDTA. These biochemical characteristics suggest that the recombinant xylanase has a prospective application in feed industry as an additive.

  16. Interleukin-1 controls the constitutive expression of the Cyp7a1 gene by regulating the expression of Cyp7a1 transcriptional regulators in the mouse liver.

    Science.gov (United States)

    Kojima, Misaki; Ashino, Takashi; Yoshida, Takemi; Iwakura, Yoichiro; Degawa, Masakuni

    2011-01-01

    Our previous study using interleukin-1α/β-knockout (IL-1-KO) and wild-type (WT) mice demonstrated that IL-1 acts as a positive factor for constitutive gene expression of hepatic cytochrome P4507a1 (Cyp7a1). In this study, to clarify the role of IL-1 in the expression of the hepatic Cyp7a1 gene, we focused on Cyp7a1 transcriptional regulators such as α-fetoprotein transcription factor (FTF), liver X receptor α (LXRα), hepatocyte nuclear factor 4α (HNF4α) and small heterodimer partner (SHP) and examined the effects of IL-1 on their gene expression by real-time reverse-transcription polymerase chain reaction using IL-1-KO and WT mice. We observed no significant differences between sex-matched IL-1-KO and WT mice with regard to gene expression levels of FTF, LXRα, and HNF4α, all of which are positive transcriptional regulators for the Cyp7a1 gene. However, interindividual differences in hepatic FTF and LXRα expression were closely dependent on the gene expression level(s) of hepatic IL-1 and tumor necrosis factor-α (TNF-α), while interindividual differences in hepatic HNF4α were clearly correlated with the expression of IL-1, but not TNF-α. In contrast, the gene expression level of SHP, which is a negative transcriptional regulator of the Cyp7a1 gene through inhibition of FTF function, was higher in IL-1-KO mice than in sex-matched WT mice. These findings demonstrate that, like TNF-α, IL-1 positively controls the gene expression of Cyp7a1 transcriptional upregulators but, in contrast to the previously reported action of TNF-α, IL-1 also acts to downregulate SHP gene expression.

  17. Establishment of Stably Transfected Cells Constitutively Expressing the Full-Length and Truncated Antigenic Proteins of Two Genetically Distinct Mink Astroviruses

    DEFF Research Database (Denmark)

    Bidokhti, Mehdi R. M.; Ullman, Karin; Jensen, Trine Hammer

    2013-01-01

    Astroviruses are becoming a growing concern in veterinary and public health. To date there are no registered vaccines against astrovirus-induced disease, mostly due to the difficulty to cultivate astroviruses to high titer for vaccine development using conventional techniques. As means to circumv......Astroviruses are becoming a growing concern in veterinary and public health. To date there are no registered vaccines against astrovirus-induced disease, mostly due to the difficulty to cultivate astroviruses to high titer for vaccine development using conventional techniques. As means...... to circumvent this drawback, we have developed stably transfected mink fetal cells and BHK21 cells constitutively expressing the full-length and truncated capsid proteins of two distinct genotypes of mink astrovirus. Protein expression in these stably transfected cells was demonstrated by strong signals...... shedding was observed in mink kits born from immunized females. The gene integration and protein expression were sustained through cell passage, showing that the used approach is robust and reliable for expression of functional capsid proteins for vaccine and diagnostic applications....

  18. Stage 3 immature human natural killer cells found in secondary lymphoid tissue constitutively and selectively express the TH17 cytokine interleukin-22

    Science.gov (United States)

    Hughes, Tiffany; Becknell, Brian; McClory, Susan; Briercheck, Edward; Freud, Aharon G.; Zhang, Xiaoli; Mao, Hsiaoyin; Nuovo, Gerard; Yu, Jianhua

    2009-01-01

    Considerable functional heterogeneity within human natural killer (NK) cells has been revealed through the characterization of distinct NK-cell subsets. Accordingly, a small subset of CD56+NKp44+NK cells, termed NK-22 cells, was recently described within secondary lymphoid tissue (SLT) as IL-22− when resting, with a minor fraction of this population becoming IL-22+ when activated. Here we discover that the vast majority of stage 3 immature NK (iNK) cells in SLT constitutively and selectively express IL-22, a TH17 cytokine important for mucosal immunity, whereas earlier and later stages of NK developmental intermediates do not express IL-22. These iNK cells have a surface phenotype of CD34−CD117+CD161+CD94−, largely lack expression of NKp44 and CD56, and do not produce IFN-γ or possess cytolytic activity. In summary, stage 3 iNK cells are highly enriched for IL-22 and IL-26 messenger RNA, and IL-22 protein production, but do not express IL-17A or IL-17F. PMID:19244159

  19. Ortho-Aminoazotoluene activates mouse Constitutive Androstane Receptor (mCAR) and increases expression of mCAR target genes

    Science.gov (United States)

    Smetanina, Mariya A.; Pakharukova, Mariya Y.; Kurinna, Svitlana M.; Dong, Bingning; Hernandez, Juan P.; Moore, David D.; Merkulova, Tatyana I.

    2011-01-01

    2'-3-dimethyl-4-aminoazobenzene (ortho-aminoazotoluene, OAT) is an azo dye and a rodent carcinogen that has been evaluated by the International Agency for Research on Cancer (IARC) as a possible (class 2B) human carcinogen. Its mechanism of action remains unclear. We examined the role of the xenobiotic receptor Constitutive Androstane Receptor (CAR, NR1I3) as a mediator of the effects of OAT. We found that OAT increases mouse CAR (mCAR) transactivation in a dose-dependent manner. This effect is specific because another closely related azo dye, 3'-methyl-4-dimethyl-aminoazobenzene (3'MeDAB), did not activate mCAR. Real-time Q-PCR analysis in wild-type C57BL/6 mice revealed that OAT induces the hepatic mRNA expression of the following CAR target genes: Cyp2b10, Cyp2c29, Cyp3a11, Ugt1a1, Mrp4, Mrp2 and c-Myc. CAR-null (Car−/−) mice showed no increased expression of these genes following OAT treatment, demonstrating that CAR is required for their OAT dependent induction. The OAT-induced CAR-dependent increase of Cyp2b10 and c-Myc expression was confirmed by Western blotting. Immunohistochemistry analysis of wild-type and Car−/− livers showed that OAT did not acutely induce hepatocyte proliferation, but at much later time points showed an unexpected CAR-dependent proliferative response. These studies demonstrate that mCAR is an OAT xenosensor, and indicate that at least some of the biological effects of this compound are mediated by this nuclear receptor. PMID:21672546

  20. Mechanical properties and constitutive relations for tantalum and tantalum alloys under high-rate deformation

    Energy Technology Data Exchange (ETDEWEB)

    Chen, S.R.; Gray, G.T. III; Bingert, S.R. [Los Alamos National Lab., NM (United States). Materials Science and Technology Div.

    1996-05-01

    Tantalum and its alloys have received increased interest as a model bcc metal and for defense-related applications. The stress-strain behavior of several tantalums, possessing varied compositions and manufacturing histories, and tantalum alloyed with tungsten, was investigated as a function of temperature from {minus}196 C to 1,000 C, and strain rate from 10{sup {minus}3} s{sup {minus}1} to 8,000 s{sup {minus}1}. The yield stress for all the Ta-materials was found to be sensitive to the test temperature, the impurity and solute contents; however, the strain hardening remained very similar for various ``pure`` tantalums but increased with alloying. Powder-metallurgy (P/M) tantalum with various levels of oxygen content produced via different processing paths was also investigated. Similar mechanical properties compared to conventionally processed tantalums were achieved in the P/M Ta. This data suggests that the frequently observed inhomogeneities in the mechanical behavior of tantalum inherited from conventional processes can be overcome. Constitutive relations based upon the Johnson-Cook, the Zerilli-Armstrong, and the Mechanical Threshold Stress models were evaluated for all the Ta-based materials. Parameters were also fit for these models to a tantalum-bar material. Flow stresses of a Ta bar stock subjected to a large-strain deformation of {var_epsilon} = 1.85 via multiple upset forging were obtained. The capabilities and limitations of each model for large-strain applications are examined. The deformation mechanisms controlling high-rate plasticity in tantalum are revisited.

  1. Characterization of a novel type of MLSB resistance plasmid from Staphylococcus saprophyticus carrying a constitutively expressed erm(C) gene.

    Science.gov (United States)

    Hauschild, Tomasz; Lüthje, Petra; Schwarz, Stefan

    2006-06-15

    An erm(C)-carrying plasmid of unusual size and restriction map, designated pSES22, was identified in a Staphylococcus saprophyticus strain and sequenced completely. Constitutive expression of the erm(C) gene from pSES22 is based on a novel 22-bp tandem duplication in the erm(C) translational attenuator. Comparative analysis of the deduced Erm(C) amino acid sequence revealed that Erm(C) from pSES22 - together with an Erm(C) methylase from S. hyicus - represented a separate branch in the homology tree of Erm(C) methylases. Structural comparisons showed that plasmid pSES22 differed distinctly from all other completely sequenced erm(C)-carrying resistance plasmids. However, pSES22 was similar to several members of a diverse group of small plasmids, all of which carried closely related plasmid backbones consisting of the genes repU and pre/mob, but differed in their resistance genes.

  2. A novel meningococcal outer membrane vesicle vaccine with constitutive expression of FetA: A phase I clinical trial.

    Science.gov (United States)

    Marsay, L; Dold, C; Green, C A; Rollier, C S; Norheim, G; Sadarangani, M; Shanyinde, M; Brehony, C; Thompson, A J; Sanders, H; Chan, H; Haworth, K; Derrick, J P; Feavers, I M; Maiden, M C; Pollard, A J

    2015-09-01

    Outer membrane vesicle (OMV) vaccines are used against outbreaks of capsular group B Neisseria meningitidis (MenB) caused by strains expressing particular PorA outer membrane proteins (OMPs). Ferric enterobactin receptor (FetA) is another variable OMP that induces type-specific bactericidal antibodies, and the combination of judiciously chosen PorA and FetA variants in vaccine formulations is a potential approach to broaden protection of such vaccines. The OMV vaccine MenPF-1 was generated by genetically modifying N. meningitidis strain 44/76 to constitutively express FetA. Three doses of 25 μg or 50 μg of MenPF-1 were delivered intra-muscularly to 52 healthy adults. MenPF-1 was safe and well tolerated. Immunogenicity was measured by serum bactericidal assay (SBA) against wild-type and isogenic mutant strains. After 3 doses, the proportion of volunteers with SBA titres ≥1:4 (the putative protective titre) was 98% for the wild-type strain, and 77% for the strain 44/76 FetA(on)PorA(off) compared to 51% in the strain 44/76 FetA(off)PorA(off), demonstrating that vaccination with MenPF-1 simultaneously induced FetA and PorA bactericidal antibodies. This study provides a proof-of-concept for generating bactericidal antibodies against FetA after OMV vaccination in humans. Prevalence-based choice of PorA and FetA types can be used to formulate a vaccine for broad protection against MenB disease. Copyright © 2015 The Authors. Published by Elsevier Ltd.. All rights reserved.

  3. Constitutive expression of fluorescent protein by Aspergillus var. niger and Aspergillus carbonarius to monitor fungal colonization in maize plants.

    Science.gov (United States)

    Palencia, Edwin Rene; Glenn, Anthony Elbie; Hinton, Dorothy Mae; Bacon, Charles Wilson

    2013-09-01

    Aspergillus niger and Aspergillus carbonarius are two species in the Aspergillus section Nigri (black-spored aspergilli) frequently associated with peanut (Arachis hypogea), maize (Zea mays), and other plants as pathogens. These infections are symptomless and as such are major concerns since some black aspergilli produce important mycotoxins, ochratoxins A, and the fumonisins. To facilitate the study of the black aspergilli-maize interactions with maize during the early stages of infections, we developed a method that used the enhanced yellow fluorescent protein (eYFP) and the monomeric red fluorescent protein (mRFP1) to transform A. niger and A. carbonarius, respectively. The results were constitutive expressions of the fluorescent genes that were stable in the cytoplasms of hyphae and conidia under natural environmental conditions. The hyphal in planta distribution in 21-day-old seedlings of maize were similar wild type and transformants of A. niger and A. carbonarius. The in planta studies indicated that both wild type and transformants internally colonized leaf, stem and root tissues of maize seedlings, without any visible disease symptoms. Yellow and red fluorescent strains were capable of invading epidermal cells of maize roots intercellularly within the first 3 days after inoculation, but intracellular hyphal growth was more evident after 7 days of inoculation. We also tested the capacity of fluorescent transformants to produce ochratoxin A and the results with A. carbonarius showed that this transgenic strain produced similar concentrations of this secondary metabolite. This is the first report on the in planta expression of fluorescent proteins that should be useful to study the internal plant colonization patterns of two ochratoxigenic species in the Aspergillus section Nigri. © 2013.

  4. Constitutive Equation for 3104 Alloy at High Temperatures in Consideration of Strain

    Science.gov (United States)

    Zhen, Fuqiang; Sun, Jianlin; Li, Jian

    2016-06-01

    The flow behavior of 3104 aluminum alloy was investigated at temperatures ranging from 250°C to 500°C, and strain rates from 0.01 to 10 s-1 by isothermal compression tests. The true stress-strain curves were obtained from the measured load-stroke data and then modified by friction and temperature correction. The effects of temperature and strain rate on hot deformation behavior were represented by Zener-Hollomon parameter including Arrhenius term. Additionally, the influence of strain was incorporated considering the effect of strain on material constants. The derived constitution equation was applied to the finite element analysis of hot compression. The results show that the simulated force is consistent with the measured one. Consequently, the developed constitution equation is valid and feasible for numerical simulation in hot deformation process of 3104 alloy.

  5. Constitutively active RAS signaling reduces 1,25 dihydroxyvitamin D-mediated gene transcription in intestinal epithelial cells by reducing vitamin D receptor expression.

    Science.gov (United States)

    DeSmet, Marsha L; Fleet, James C

    2017-10-01

    High vitamin D status is associated with reduced colon cancer risk but these studies ignore the diversity in the molecular etiology of colon cancer. RAS activating mutations are common in colon cancer and they activate pro-proliferative signaling pathways. We examined the impact of RAS activating mutations on 1,25 dihydroxyvitamin D (1,25(OH)2D)-mediated gene expression in cultured colon and intestinal cell lines. Transient transfection of Caco-2 cells with a constitutively active mutant K-RAS (G12 V) significantly reduced 1,25(OH)2D-induced activity of both a human 25-hydroxyvitamin D, 24 hydroxyase (CYP24A1) promoter-luciferase and an artificial 3X vitamin D response element (VDRE) promoter-luciferase reporter gene. Young Adult Mouse Colon (YAMC) and Rat Intestinal Epithelial (RIE) cell lines with stable expression of mutant H-RAS had suppressed 1,25(OH)2D-mediated induction of CYP24A1 mRNA. The RAS effects were associated with lower Vitamin D receptor (VDR) mRNA and protein levels in YAMC and RIE cells and they could be partially reversed by VDR overexpression. RAS-mediated suppression of VDR levels was not due to either reduced VDR mRNA stability or increased VDR gene methylation. However, chromatin accessibility to the VDR gene at the proximal promoter (-300bp), an enhancer region at -6kb, and an enhancer region located in exon 3 was significantly reduced in RAS transformed YAMC cells (YAMC-RAS). These data show that constitutively active RAS signaling suppresses 1,25(OH)2D-mediated gene transcription in colon epithelial cells by reducing VDR gene transcription but the mechanism for this suppression is not yet known. These data suggest that cancers with RAS-activating mutations may be less responsive to vitamin D mediated treatment or chemoprevention. Copyright © 2017 Elsevier Ltd. All rights reserved.

  6. Constitutive Models

    DEFF Research Database (Denmark)

    2011-01-01

    This chapter presents various types of constitutive models and their applications. There are 3 aspects dealt with in this chapter, namely: creation and solution of property models, the application of parameter estimation and finally application examples of constitutive models. A systematic...

  7. Up-regulation of phosphoinositide metabolism in tobacco cells constitutively expressing the human type I inositol polyphosphate 5-phosphatase

    Science.gov (United States)

    Perera, Imara Y.; Love, John; Heilmann, Ingo; Thompson, William F.; Boss, Wendy F.; Brown, C. S. (Principal Investigator)

    2002-01-01

    To evaluate the impact of suppressing inositol 1,4,5-trisphosphate (InsP(3)) in plants, tobacco (Nicotiana tabacum) cells were transformed with the human type I inositol polyphosphate 5-phosphatase (InsP 5-ptase), an enzyme which specifically hydrolyzes InsP(3). The transgenic cell lines showed a 12- to 25-fold increase in InsP 5-ptase activity in vitro and a 60% to 80% reduction in basal InsP(3) compared with wild-type cells. Stimulation with Mas-7, a synthetic analog of the wasp venom peptide mastoparan, resulted in an approximately 2-fold increase in InsP(3) in both wild-type and transgenic cells. However, even with stimulation, InsP(3) levels in the transgenic cells did not reach wild-type basal values, suggesting that InsP(3) signaling is compromised. Analysis of whole-cell lipids indicated that phosphatidylinositol 4,5-bisphosphate (PtdInsP(2)), the lipid precursor of InsP(3), was greatly reduced in the transgenic cells. In vitro assays of enzymes involved in PtdInsP(2) metabolism showed that the activity of the PtdInsP(2)-hydrolyzing enzyme phospholipase C was not significantly altered in the transgenic cells. In contrast, the activity of the plasma membrane PtdInsP 5 kinase was increased by approximately 3-fold in the transgenic cells. In vivo labeling studies revealed a greater incorporation of (32)P into PtdInsP(2) in the transgenic cells compared with the wild type, indicating that the rate of PtdInsP(2) synthesis was increased. These studies show that the constitutive expression of the human type I InsP 5-ptase in tobacco cells leads to an up-regulation of the phosphoinositide pathway and highlight the importance of PtdInsP(2) synthesis as a regulatory step in this system.

  8. Postnatal expression in hyaline cartilage of constitutively active human collagenase-3 (MMP-13) induces osteoarthritis in mice

    National Research Council Canada - National Science Library

    Neuhold, L A; Killar, L; Zhao, W; Sung, M L; Warner, L; Kulik, J; Turner, J; Wu, W; Billinghurst, C; Meijers, T; Poole, A R; Babij, P; DeGennaro, L J

    2001-01-01

    ...). We have used tetracycline-regulated transcription in conjunction with a cartilage-specific promoter to target a constitutively active human MMP-13 to the hyaline cartilages and joints of transgenic mice...

  9. Mechanical properties and constitutive relations for molybdenum under high-rate deformation

    Energy Technology Data Exchange (ETDEWEB)

    Chen, S.R.; Maudlin, P.J.; Gray, G.T. III

    1998-01-01

    Molybdenum and its alloys have received increased interest in recent years for ballistic applications. The stress-strain behavior of several molybdenums possessing various compositions, manufacturing sources, and the degree of pre-straining, were investigated as a function of temperature from 77 to 1,273 K, and strain rate from 10{sup {minus}3} s{sup {minus}1} to 8,000 s{sup {minus}1}. The yield stress was found to be sensitive to the test temperature and strain rate, however, the strain hardening remained rate-insensitive. The constitutive response of a powder-metallurgy molybdenum was also investigated; similar mechanical properties compared to conventionally wrought processed molybdenums were achieved. Constitutive relations based upon the Johnson-Cook, the Zerilli-Armstrong and the Mechanical Threshold Stress (MTS) models were evaluated and fit for the various Mo-based materials. The capabilities and limitations of each model for large-strain applications were examined. The differences between the three models are demonstrated using model comparisons to Taylor cylinder validation experiments.

  10. The effect of heat developed during high strain rate deformation on the constitutive modeling of amorphous polymers

    Science.gov (United States)

    Safari, Keivan H.; Zamani, Jamal; Guedes, Rui M.; Ferreira, Fernando J.

    2016-02-01

    An adiabatic constitutive model is proposed for large strain deformation of polycarbonate (PC) at high strain rates. When the strain rate is sufficiently high such that the heat generated does not have time to transfer to the surroundings, temperature of material rises. The high strain rate deformation behavior of polymers is significantly affected by temperature-dependent constants and thermal softening. Based on the isothermal model which first was introduced by Mulliken and Boyce et al. (Int. J. Solids Struct. 43:1331-1356, 2006), an adiabatic model is proposed to predict the yield and post-yield behavior of glassy polymers at high strain rates. When calculating the heat generated and the temperature changes during the step by step simulation of the deformation, temperature-dependent elastic constants are incorporated to the constitutive equations. Moreover, better prediction of softening phenomena is achieved by the new definition for softening parameters of the proposed model. The constitutive model has been implemented numerically into a commercial finite element code through a user material subroutine (VUMAT). The experimental results, obtained using a split Hopkinson pressure bar, are supported by dynamic mechanical thermal analysis (DMTA) and Decompose/Shift/Reconstruct (DSR) method. Comparison of adiabatic model predictions with experimental data demonstrates the ability of the model to capture the characteristic features of stress-strain curve of the material at very high strain rates.

  11. Construction and characterization of stable, constitutively expressed, chromosomal green and red fluorescent transcriptional fusions in the select agents, Bacillus anthracis, Yersinia pestis, Burkholderia mallei, and Burkholderia pseudomallei.

    Science.gov (United States)

    Su, Shengchang; Bangar, Hansraj; Saldanha, Roland; Pemberton, Adin; Aronow, Bruce; Dean, Gary E; Lamkin, Thomas J; Hassett, Daniel J

    2014-10-01

    Here, we constructed stable, chromosomal, constitutively expressed, green and red fluorescent protein (GFP and RFP) as reporters in the select agents, Bacillus anthracis, Yersinia pestis, Burkholderia mallei, and Burkholderia pseudomallei. Using bioinformatic approaches and other experimental analyses, we identified P0253 and P1 as potent promoters that drive the optimal expression of fluorescent reporters in single copy in B. anthracis and Burkholderia spp. as well as their surrogate strains, respectively. In comparison, Y. pestis and its surrogate strain need two chromosomal copies of cysZK promoter (P2cysZK) for optimal fluorescence. The P0253-, P2cysZK-, and P1-driven GFP and RFP fusions were first cloned into the vectors pRP1028, pUC18R6KT-mini-Tn7T-Km, pmini-Tn7-gat, or their derivatives. The resultant constructs were delivered into the respective surrogates and subsequently into the select agent strains. The chromosomal GFP- and RFP-tagged strains exhibited bright fluorescence at an exposure time of less than 200 msec and displayed the same virulence traits as their wild-type parental strains. The utility of the tagged strains was proven by the macrophage infection assays and lactate dehydrogenase release analysis. Such strains will be extremely useful in high-throughput screens for novel compounds that could either kill these organisms, or interfere with critical virulence processes in these important bioweapon agents and during infection of alveolar macrophages.

  12. Differential expression profiles and roles of inducible DUSPs and ERK1/2-specific constitutive DUSP6 and DUSP7 in microglia.

    Science.gov (United States)

    Ham, Ji-Eun; Oh, Eun-Kyung; Kim, Dong-Hoon; Choi, Sang-Hyun

    2015-11-13

    Dual-specificity phosphatases (DUSPs) show distinct substrate preferences for specific MAPKs. DUSPs sharing a substrate preference for ERK1/2 may be classified as inducible or constitutive. In contrast to the inducible DUSPs which also dephosphorylate p38 MAPK and JNK in the major inflammatory pathways, constitutive DUSP6 and DUSP7 are specific to ERK1/2 and have not been studied in microglia and other immune cells to date. In the present study, we differentiated mRNA expression profiles of inducible and constitutive DUSPs that dephosphorylate ERK1/2 in microglia. Lipopolysaccharide (LPS) at 1 ng/ml induced prompt phosphorylation of ERK1/2 with peak induction at 30 min. LPS induced expression of DUSP1, DUSP2, and DUSP5 within 60 min, whereas DUSP4 expression was induced more slowly. DUSP6 and DUSP7 exhibited constitutive basal expression, which decreased immediately after LPS stimulation but subsequently returned to basal levels. The expression of DUSP6 and DUSP7 was regulated inverse to the phosphorylation of ERK1/2 in LPS-stimulated microglia. Therefore, we next investigated the correlation between DUSP6 and DUSP7 expression and ERK1/2 phosphorylation in resting and LPS-stimulated microglia. Inhibition of the ERK1/2 pathway by PD98059 and FR180204 resulted in a decrease in DUSP6 and DUSP7 expression, both in resting and LPS-stimulated microglia. These inhibitors partially blocked the LPS-induced expression of DUSP1, DUSP2, and DUSP4, but had no effect on DUSP5. Finally, we examined the role of DUSP6 activity in the downregulation of ERK1/2 phosphorylation. BCI, an inhibitor of DUSP6, increased the phosphorylation of ERK1/2. However, pretreatment with BCI inhibited the LPS-induced phosphorylation of ERK1/2. These results demonstrate that constitutive DUPS6 and DUSP7 expression was downregulated inverse to the expression of inducible DUSPs and the phosphorylation of ERK1/2 in LPS-stimulated microglia. The expression of DUPS6 and DUSP7 was mediated by ERK1

  13. Developmental Constitutionalism

    OpenAIRE

    Skupien, Stefan

    2015-01-01

    The search for adequate political and socio-economic models continues within recent constitutional reforms in Sub-Saharan post-colonial societies since 2005. This discourse goes back to the period of decolonisation but also to the transitions after 1989. Within this study, I assess the problem of representational crisis that come hand in hand with constitution making processes. Especially, I focus on the representation of different groups within the state, the safeguarding and extent of basic...

  14. Constitutive expression of a COOH-terminal leucine mutant of lysosome-associated membrane protein-1 causes its exclusive localization in low density intracellular vesicles.

    Science.gov (United States)

    Akasaki, Kenji; Shiotsu, Keiko; Michihara, Akihiro; Ide, Norie; Wada, Ikuo

    2014-07-01

    Lysosome-associated membrane protein-1 (LAMP-1) is a type I transmembrane protein with a short cytoplasmic tail that possesses a lysosome-targeting signal of GYQTI(382)-COOH. Wild-type (WT)-LAMP-1 was exclusively localized in high density lysosomes, and efficiency of LAMP-1's transport to lysosomes depends on its COOH-terminal amino acid residue. Among many different COOH-terminal amino acid substitution mutants of LAMP-1, a leucine-substituted mutant (I382L) displays the most efficient targeting to late endosomes and lysosomes [Akasaki et al. (2010) J. Biochem. 148: , 669-679]. In this study, we generated two human hepatoma cell lines (HepG2 cell lines) that stably express WT-LAMP-1 and I382L, and compared their intracellular distributions. The subcellular fractionation study using Percoll density gradient centrifugation revealed that WT-LAMP-1 had preferential localization in the high density secondary lysosomes where endogenous human LAMP-1 was enriched. In contrast, a major portion of I382L was located in a low density fraction. The low density fraction also contained approximately 80% of endogenous human LAMP-1 and significant amounts of endogenous β-glucuronidase and LAMP-2, which probably represents occurrence of low density lysosomes in the I382L-expressing cells. Double immunofluorescence microscopic analyses distinguished I382L-containing intracellular vesicles from endogenous LAMP-1-containing lysosomes and early endosomes. Altogether, constitutive expression of I382L causes its aberrant intracellular localization and generation of low density lysosomes, indicating that the COOH-terminal isoleucine is critical for normal localization of LAMP-1 in the dense lysosomes.

  15. CONSTITUTIVE MODEL OF STEEL FIBRE REINFORCED CONCRETE SUBJECTED TO HIGH TEMPERATURES

    Directory of Open Access Journals (Sweden)

    Lukas Blesak

    2016-12-01

    Full Text Available Research on structural load-bearing systems exposed to elevated temperatures is an active topic in civil engineering. Carrying out a full-size experiment of a specimen exposed to fire is a challenging task considering not only the preparation labour but also the necessary costs. Therefore, such experiments are simulated using various software and computational models in order to predict the structural behaviour as exactly as possible. In this paper such a procedure, focusing on software simulation, is described in detail. The proposed constitutive model is based on the stress-strain curve and allows predicting SFRC material behaviour in bending at ambient and elevated temperature. SFRC material is represented by the initial linear behaviour, an instantaneous drop of stress after the initial crack occurs and its consequent specific ductility, which influences the overall modelled specimen behaviour under subjected loading. The model is calibrated with ATENA FEM software using experimental results.

  16. Constitutive or Inducible Protective Mechanisms against UV-B Radiation in the Brown Alga Fucus vesiculosus? A Study of Gene Expression and Phlorotannin Content Responses.

    Directory of Open Access Journals (Sweden)

    Emeline Creis

    Full Text Available A role as UV sunscreens has been suggested for phlorotannins, the phenolic compounds that accumulate in brown algae in response to a number of external stimuli and take part in cell wall structure. After exposure of the intertidal brown alga Fucus vesiculosus to artificial UV-B radiation, we examined its physiological responses by following the transcript level of the pksIII gene encoding a phloroglucinol synthase, likely to be involved in the first step of phlorotannins biosynthesis. We also monitored the expression of three targeted genes, encoding a heat shock protein (hsp70, which is involved in global stress responses, an aryl sulfotransferase (ast, which could be involved in the sulfation of phlorotannins, and a vanadium bromoperoxidase (vbpo, which can potentially participate in the scavenging of Reactive Oxygen Species (ROS and in the cross-linking and condensation of phlorotannins. We investigated whether transcriptional regulation of these genes is correlated with an induction of phlorotannin accumulation by establishing metabolite profiling of purified fractions of low molecular weight phlorotannins. Our findings demonstrated that a high dose of UV-B radiation induced a significant overexpression of hsp70 after 12 and 24 hours following the exposure to the UV-B treatment, compared to control treatment. The physiological performance of algae quantified by the photosynthetic efficiency (Fv/Fm was slightly reduced. However UV-B treatment did not induce the accumulation of soluble phlorotannins in F. vesiculosus during the kinetics of four weeks, a result that may be related to the lack of induction of the pksIII gene expression. Taken together these results suggest a constitutive accumulation of phlorotannins occurring during the development of F.vesiculosus, rather than inducible processes. Gene expression studies and phlorotannin profiling provide here complementary approaches to global quantifications currently used in studies of

  17. Constitutive or Inducible Protective Mechanisms against UV-B Radiation in the Brown Alga Fucus vesiculosus? A Study of Gene Expression and Phlorotannin Content Responses.

    Science.gov (United States)

    Creis, Emeline; Delage, Ludovic; Charton, Sophie; Goulitquer, Sophie; Leblanc, Catherine; Potin, Philippe; Ar Gall, Erwan

    2015-01-01

    A role as UV sunscreens has been suggested for phlorotannins, the phenolic compounds that accumulate in brown algae in response to a number of external stimuli and take part in cell wall structure. After exposure of the intertidal brown alga Fucus vesiculosus to artificial UV-B radiation, we examined its physiological responses by following the transcript level of the pksIII gene encoding a phloroglucinol synthase, likely to be involved in the first step of phlorotannins biosynthesis. We also monitored the expression of three targeted genes, encoding a heat shock protein (hsp70), which is involved in global stress responses, an aryl sulfotransferase (ast), which could be involved in the sulfation of phlorotannins, and a vanadium bromoperoxidase (vbpo), which can potentially participate in the scavenging of Reactive Oxygen Species (ROS) and in the cross-linking and condensation of phlorotannins. We investigated whether transcriptional regulation of these genes is correlated with an induction of phlorotannin accumulation by establishing metabolite profiling of purified fractions of low molecular weight phlorotannins. Our findings demonstrated that a high dose of UV-B radiation induced a significant overexpression of hsp70 after 12 and 24 hours following the exposure to the UV-B treatment, compared to control treatment. The physiological performance of algae quantified by the photosynthetic efficiency (Fv/Fm) was slightly reduced. However UV-B treatment did not induce the accumulation of soluble phlorotannins in F. vesiculosus during the kinetics of four weeks, a result that may be related to the lack of induction of the pksIII gene expression. Taken together these results suggest a constitutive accumulation of phlorotannins occurring during the development of F.vesiculosus, rather than inducible processes. Gene expression studies and phlorotannin profiling provide here complementary approaches to global quantifications currently used in studies of phenolic compounds

  18. Constitutive CCND1/CDK2 activity substitutes for p53 loss, or MYC or oncogenic RAS expression in the transformation of human mammary epithelial cells.

    Directory of Open Access Journals (Sweden)

    Damian J Junk

    Full Text Available Cancer develops following the accumulation of genetic and epigenetic alterations that inactivate tumor suppressor genes and activate proto-oncogenes. Dysregulated cyclin-dependent kinase (CDK activity has oncogenic potential in breast cancer due to its ability to inactivate key tumor suppressor networks and drive aberrant proliferation. Accumulation or over-expression of cyclin D1 (CCND1 occurs in a majority of breast cancers and over-expression of CCND1 leads to accumulation of activated CCND1/CDK2 complexes in breast cancer cells. We describe here the role of constitutively active CCND1/CDK2 complexes in human mammary epithelial cell (HMEC transformation. A genetically-defined, stepwise HMEC transformation model was generated by inhibiting p16 and p53 with shRNA, and expressing exogenous MYC and mutant RAS. By replacing components of this model, we demonstrate that constitutive CCND1/CDK2 activity effectively confers anchorage independent growth by inhibiting p53 or replacing MYC or oncogenic RAS expression. These findings are consistent with several clinical observations of luminal breast cancer sub-types that show elevated CCND1 typically occurs in specimens that retain wild-type p53, do not amplify MYC, and contain no RAS mutations. Taken together, these data suggest that targeted inhibition of constitutive CCND1/CDK2 activity may enhance the effectiveness of current treatments for luminal breast cancer.

  19. Constitutive Modeling of High-Temperature Flow Behavior of an Nb Micro-alloyed Hot Stamping Steel

    Science.gov (United States)

    Zhang, Shiqi; Feng, Ding; Huang, Yunhua; Wei, Shizhong; Mohrbacher, Hardy; Zhang, Yue

    2016-03-01

    The thermal deformation behavior and constitutive models of an Nb micro-alloyed 22MnB5 steel were investigated by conducting isothermal uniaxial tensile tests at the temperature range of 873-1223 K with strain rates of 0.1-10 s-1. The results indicated that the investigated steel showed typical work hardening and dynamic recovery behavior during hot deformation, and the flow stress decreased with a decrease in strain rate and/or an increase in temperature. On the basis of the experimental data, the modified Johnson-Cook (modified JC), modified Norton-Hoff (modified NH), and Arrhenius-type (AT) constitutive models were established for the subject steel. However, the flow stress values predicted by these three models revealed some remarkable deviations from the experimental values for certain experimental conditions. Therefore, a new combined modified Norton-Hoff and Arrhenius-type constitutive model (combined modified NH-AT model), which accurately reflected both the work hardening and dynamic recovery behavior of the subject steel, was developed by introducing the modified parameter k ɛ. Furthermore, the accuracy of these constitutive models was assessed by the correlation coefficient, the average absolute relative error, and the root mean square error, which indicated that the flow stress values computed by the combined modified NH-AT model were highly consistent with the experimental values (R = 0.998, AARE = 1.63%, RMSE = 3.85 MPa). The result confirmed that the combined modified NH-AT model was suitable for the studied Nb micro-alloyed hot stamping steel. Additionally, the practicability of the new model was also verified using finite element simulations in ANSYS/LS-DYNA, and the results confirmed that the new model was practical and highly accurate.

  20. Expression of a constitutively active calcineurin encoded by an intron-retaining mRNA in follicular keratinocytes.

    Directory of Open Access Journals (Sweden)

    Atsushi Fujimura

    Full Text Available Hair growth is a highly regulated cyclical process. Immunosuppressive immunophilin ligands such as cyclosporin A (CsA and FK506 are known as potent hair growth modulatory agents in rodents and humans that induce active hair growth and inhibit hair follicle regression. The immunosuppressive effectiveness of these drugs has been generally attributed to inhibition of T cell activation through well-characterized pathways. Specifically, CsA and FK506 bind to intracellular proteins, principally cyclophilin A and FKBP12, respectively, and thereby inhibit the phosphatase calcineurin (Cn. The calcineurin (Cn/NFAT pathway has an important, but poorly understood, role in the regulation of hair follicle development. Here we show that a novel-splicing variant of calcineurin Aß CnAß-FK, which is encoded by an intron-retaining mRNA and is deficient in the autoinhibitory domain, is predominantly expressed in mature follicular keratinocytes but not in the proliferating keratinocytes of rodents. CnAß-FK was weakly sensitive to Ca(2+ and dephosphorylated NFATc2 under low Ca(2+ levels in keratinocytes. Inhibition of Cn/NFAT induced hair growth in nude mice. Cyclin G2 was identified as a novel target of the Cn/NFATc2 pathway and its expression in follicular keratinocytes was reduced by inhibition of Cn/NFAT. Overexpression of cyclin G2 arrested the cell cycle in follicular keratinocytes in vitro and the Cn inhibitor, cyclosporin A, inhibited nuclear localization of NFATc2, resulting in decreased cyclin G2 expression in follicular keratinocytes of rats in vivo. We therefore suggest that the calcineurin/NFAT pathway has a unique regulatory role in hair follicle development.

  1. Constitutional Conservatism

    Science.gov (United States)

    Berkowitz, Peter

    2009-01-01

    After their dismal performance in election 2008, conservatives are taking stock. As they examine the causes that have driven them into the political wilderness and as they explore paths out, they should also take heart. After all, election 2008 shows that America's constitutional order is working as designed. Indeed, while sorting out their errors…

  2. A constitutive model for the anelastic behavior of Advanced High Strength Steels

    NARCIS (Netherlands)

    Torkabadi, A.; Liempt, van P.; Meinders, V.T.; Boogaard, van den A.H.

    2015-01-01

    In this work a physically based model describing the anelastic behaviour and nonlinear unloading in Advanced High Strength Steels (AHSS) is proposed. The model is fitted to the experimental data obtained from uni-axial tests on a dual-phase high strength steel grade (HCT780). The results show a good

  3. Altered methylation and expression of ER-associated degradation factors in long-term alcohol and constitutive ER stress-induced murine hepatic tumors

    Directory of Open Access Journals (Sweden)

    Hui eHan

    2013-10-01

    Full Text Available Mortality from liver cancer in humans is increasingly attributable to heavy or long-term alcohol consumption. The mechanisms by which alcohol exerts its carcinogenic effect are not well understood. In this study, the role of alcohol-induced endoplasmic reticulum (ER stress response in liver cancer development was investigated using an animal model with a liver knockout of the chaperone BiP and under constitutive hepatic ER stress. Long-term alcohol and high fat diet (HFD feeding resulted in higher levels of serum alanine aminotransferase (ALT, impaired ER stress response, and higher incidence of liver tumor in older (aged 16 months knockout females than in either middle-aged (6 months knockouts or older (aged 16 months wild type females. In the older knockout females, stronger effects of the alcohol on methylation of CpG islands at promoter regions of genes involved in the ER associated degradation (ERAD were also detected. Altered expression of ERAD factors including derlin 3, Creld2 (cysteine-rich with EGF-like domains 2, Herpud1 (ubiquitin-like domain member, Wfs1 (wolfram syndrome gene, and Yod1 (deubiquinating enzyme 1 was co-present with decreased proteasome activities, increased estrogen receptor alpha variant (ERa36, and enhanced phosphorylations of ERK1/2 (extracellular signal-regulated protein kinases 1 and 2 and STAT3 (the signal transducers and activators of transcription in the older knockout female fed alcohol. Our results suggest that long-term alcohol consumption and ageing may promote liver tumorigenesis in females through interfering with DNA methylation and expression of genes involved in the ER associated degradation.

  4. Constitutive expression of pathogen-inducible OsWRKY31 enhances disease resistance and affects root growth and auxin response in transgenic rice plants

    Institute of Scientific and Technical Information of China (English)

    Juan Zhang; Youliang Peng; Zejian Guo

    2008-01-01

    WRKY transcription factors have many regulatory roles in response to biotic and abiotic stresses. In this study, we isolated a rice WRKY gene (OsWRKY31) that is induced by the rice blast fungus Magnaporthe grisea and auxin. This gene encodes a polypeptide of 211 amino-acid residues and belongs to a subgroup of the rice WRKY gene family that probably originated after the divergence of monocot and dicot plants. OsWRKY31 was found to be localized to the nucleus of onion epidermis cells to transiently express OsWRKY31-eGFP fusion protein. Analysis of 0sWRKY31 and its mutants fused with a Cal4 DNA-binding domain indicated that OsWRKY31 has transactivation activity in yeast. Overexpression of the OsWRKY31 gene was found to enhance resistance against infection with M. grisea, and the transgenic lines exhibited reduced lateral root formation and elongation compared with wild-type and RNAi plants. The lines with overexpression showed constitutive expression of many defense-related genes, such as PBZ1 and OsSci2, as well as early auxin-response genes, such as OsIAA4 and OsCrll genes. Furthermore, the plants with overexpression were less sensitive to exogenously supplied IBA, NAA and 2,4-D at high concentrations, suggesting that overexpression of the OsWRKY31 gene might alter the auxin response or transport. These results also suggest that OsWRKY31 might be a common component in the signal transduction pathways of the auxin response and the defense response in rice.

  5. High-level expression of a chemically synthesized gene for human interferon-gamma using a prokaryotic expression vector.

    OpenAIRE

    1984-01-01

    A chemically synthesized gene for human interferon-gamma has been cloned into a prokaryotic expression vector under the regulation of a synthetic constitutive transcriptional-translational control unit that contains a strong bacteriophage T5 early promoter and a strong ribosome-binding site. Cells harboring the recombinant plasmid express high levels (4 X 10(9) units per liter of culture) of antiviral activity specific for interferon-gamma. Analysis of total cell lysates on NaDodSO4/polyacryl...

  6. Constitutive Models

    DEFF Research Database (Denmark)

    2011-01-01

    procedure is introduced for the analysis and solution of property models. Models that capture and represent the temperature dependent behaviour of physical properties are introduced, as well as equation of state models (EOS) such as the SRK EOS. Modelling of liquid phase activity coefficients are also......This chapter presents various types of constitutive models and their applications. There are 3 aspects dealt with in this chapter, namely: creation and solution of property models, the application of parameter estimation and finally application examples of constitutive models. A systematic...... covered, illustrating several models such as the Wilson equation and NRTL equation, along with their solution strategies. A section shows how to use experimental data to regress the property model parameters using a least squares approach. A full model analysis is applied in each example that discusses...

  7. Constitutive expression of IL-18 and IL-18R in differentiated IEC-6 cells: effect of TNF-alpha and IFN-gamma treatment.

    Science.gov (United States)

    Kolinska, Jirina; Lisa, Vera; Clark, Jessica A; Kozakova, Hana; Zakostelecka, Marie; Khailova, Ludmila; Sinkora, Marek; Kitanovicova, Andrea; Dvorak, Bohuslav

    2008-05-01

    The multifunctional cytokine interleukin-18 (IL-18) is an important mediator in intestinal inflammatory processes. The aim of this study was to evaluate the constitutive expression of IL-18 and its receptors (IL-18Ralpha and IL-18Rbeta) in intestinal epithelial cells (IEC) stimulated by tumor necrosis factor-alpha (TNF-alpha) and interferon-gamma (IFN-gamma). In addition, cellular proliferation and evaluation of brush border enzymes as differentiation markers were studied. Nontransformed rat intestinal epithelial IEC-6 cells were grown on an extracellular matrix (ECM) in medium with or without TNF-alpha, IFN-gamma, or a combination of both. Gene expression of IL-18, its receptors and apoptotic markers was evaluated using real-time PCR. Expression of IL-18Ralpha protein was demonstrated by flow cytometry and Western blot. Enzymatic activities of brush border enzymes and caspase-1 were determined. The constitutive expression of IL-18, IL-18Ralpha and IL-18Rbeta mRNAs and proteins were detected in IEC-6 cells. The biologically active form of IL-18 was released in response to TNF-alpha and IFN-gamma treatment. Exogenous IL-18 had no effect on cellular proliferation, brush border enzyme activities, and gene expression of apoptotic markers. However, the addition of IL-18 stimulated production and release of the chemokine IL-8. These data suggest that IEC-6 cells may be not only a source of IL-18 but also a target for its action.

  8. FetA Antibodies Induced by an Outer Membrane Vesicle Vaccine Derived from a Serogroup B Meningococcal Isolate with Constitutive FetA Expression.

    Directory of Open Access Journals (Sweden)

    Holly Sanders

    Full Text Available Invasive meningococcal disease causes over 3500 cases each year in Europe, with particularly high incidence among young children. Among serogroup B meningococci, which cause most of the cases, high diversity in the outer membrane proteins (OMPs is observed in endemic situations; however, comprehensive molecular epidemiological data are available for the diversity and distribution of the OMPs PorA and FetA and these can be used to rationally design a vaccine with high coverage of the case isolates. The aim of this study was to determine whether outer membrane vesicles (OMVs derived from an isolate with constitutive FetA expression (MenPF-1 vaccine could be used to induce antibodies against both the PorA and FetA antigens. The immunogenicity of various dose levels and number of doses was evaluated in mice and rabbits, and IgG antibody responses tested against OMVs and recombinant PorA and FetA proteins. A panel of four isogenic mutants was generated and used to evaluate the relative ability of the vaccine to induce serum bactericidal activity (SBA against FetA and PorA. Sera from mice were tested in SBA against the four target strains. Results demonstrated that the MenPF-1 OMVs were immunogenic against PorA and FetA in both animal models. Furthermore, the murine antibodies induced were bactericidal against isogenic mutant strains, suggesting that antibodies to both PorA and FetA were functional. The data presented indicate that the MenPF-1 vaccine is a suitable formulation for presenting PorA and FetA OMPs in order to induce bactericidal antibodies, and that proceeding to a Phase I clinical trial with this vaccine candidate is justified.

  9. FetA Antibodies Induced by an Outer Membrane Vesicle Vaccine Derived from a Serogroup B Meningococcal Isolate with Constitutive FetA Expression.

    Science.gov (United States)

    Sanders, Holly; Norheim, Gunnstein; Chan, Hannah; Dold, Christina; Vipond, Caroline; Derrick, Jeremy P; Pollard, Andrew J; Maiden, Martin C J; Feavers, Ian M

    2015-01-01

    Invasive meningococcal disease causes over 3500 cases each year in Europe, with particularly high incidence among young children. Among serogroup B meningococci, which cause most of the cases, high diversity in the outer membrane proteins (OMPs) is observed in endemic situations; however, comprehensive molecular epidemiological data are available for the diversity and distribution of the OMPs PorA and FetA and these can be used to rationally design a vaccine with high coverage of the case isolates. The aim of this study was to determine whether outer membrane vesicles (OMVs) derived from an isolate with constitutive FetA expression (MenPF-1 vaccine) could be used to induce antibodies against both the PorA and FetA antigens. The immunogenicity of various dose levels and number of doses was evaluated in mice and rabbits, and IgG antibody responses tested against OMVs and recombinant PorA and FetA proteins. A panel of four isogenic mutants was generated and used to evaluate the relative ability of the vaccine to induce serum bactericidal activity (SBA) against FetA and PorA. Sera from mice were tested in SBA against the four target strains. Results demonstrated that the MenPF-1 OMVs were immunogenic against PorA and FetA in both animal models. Furthermore, the murine antibodies induced were bactericidal against isogenic mutant strains, suggesting that antibodies to both PorA and FetA were functional. The data presented indicate that the MenPF-1 vaccine is a suitable formulation for presenting PorA and FetA OMPs in order to induce bactericidal antibodies, and that proceeding to a Phase I clinical trial with this vaccine candidate is justified.

  10. Development of wide-range constitutive equations for calculations of high-rate deformation of metals

    Directory of Open Access Journals (Sweden)

    Preston D.

    2011-01-01

    Full Text Available For development of models of strength and compressibility of metals in wide range of pressures (up to several megabar and strain rates ~ 1÷108 s−1, the method of dynamic tests is used. Since direct measurement of strength is impossible under complicated intensive high-rate loading, a formal model is created at first, and then it is updated basing on comparison with many experiments, which are sensitive to shear strength. Elastic-plastic, viscous-elastic-plastic and relaxation integral models became nowadays most commonly used. The basic unsolved problems in simulation of high-rate deformation of metals are mentioned in the paper.

  11. At High Levels, Constitutively Activated STAT3 Induces Apoptosis of Chronic Lymphocytic Leukemia Cells.

    Science.gov (United States)

    Rozovski, Uri; Harris, David M; Li, Ping; Liu, Zhiming; Wu, Ji Yuan; Grgurevic, Srdana; Faderl, Stefan; Ferrajoli, Alessandra; Wierda, William G; Martinez, Matthew; Verstovsek, Srdan; Keating, Michael J; Estrov, Zeev

    2016-05-15

    In chronic lymphocytic leukemia (CLL), the increment in PBLs is slower than the expected increment calculated from the cells' proliferation rate, suggesting that cellular proliferation and apoptosis are concurrent. Exploring this phenomenon, we found overexpression of caspase-3, higher cleaved poly (ADP-ribose) polymerase levels (p < 0.007), and a higher apoptosis rate in cells from patients with high counts compared with cells from patients with low counts. Although we previously found that STAT3 protects CLL cells from apoptosis, STAT3 levels were significantly higher in cells from patients with high counts than in cells from patients with low counts. Furthermore, overexpression of STAT3 did not protect the cells. Rather, it upregulated caspase-3 and induced apoptosis. Remarkably, putative STAT3 binding sites were identified in the caspase-3 promoter, and a luciferase assay, chromatin immunoprecipitation, and an EMSA revealed that STAT3 activated caspase-3 However, caspase-3 levels increased only when STAT3 levels were sufficiently high. Using chromatin immunoprecipitation and EMSA, we found that STAT3 binds with low affinity to the caspase-3 promoter, suggesting that at high levels, STAT3 activates proapoptotic mechanisms and induces apoptosis in CLL cells.

  12. What Constitutes High-Quality Discussion in Science? Research from the Perspectives on Science Course

    Science.gov (United States)

    Levinson, Ralph; Hand, Michael; Amos, Ruth

    2012-01-01

    Perspectives on Science (POS) is a unique research-based post-16 course that addresses the history, philosophy and ethical aspects of science. Our central research question was to what extent is POS successful in promoting high-quality discussion in class and what factors influence this. Through questionnaires, interviews and observations of…

  13. Inducible but not constitutive expression of PD-L1 in human melanoma cells is dependent on activation of NF-κB.

    Directory of Open Access Journals (Sweden)

    Kavitha Gowrishankar

    Full Text Available Monoclonal antibodies against immune checkpoint blockade have proven to be a major success in the treatment of melanoma. The programmed death receptor-1 ligand-1 (PD-L1 expression on melanoma cells is believed to have an inhibitory effect on T cell responses and to be an important escape mechanism from immune attack. Previous studies have shown that PD-L1 can be expressed constitutively or can be induced by IFN-γ secreted by infiltrating lymphocytes. In the present study we have investigated the mechanism underlying these two modes of PD-L1 expression in melanoma cells including cells that had acquired resistance to the BRAF inhibitor vemurafenib. PD-L1 expression was examined by flow cytometry and immunoblotting. Specific inhibitors and siRNA knockdown approaches were used to examine the roles of the RAF/ MEK, PI3K, NF-κB, STAT3 and AP1/ c-Jun pathways. IFN-γ inducible expression of PD-L1 was dependent on NF-κB as shown by inhibition with BMS-345541, an inhibitor of IκB and the BET protein inhibitor I-BET151, as well as by siRNA knockdown of NF-κB subunits. We were unable to implicate the BRAF/MEK pathway as major regulators in PD-L1 expression on vemurafenib resistant cells. Similarly the PI3K/AKT pathway and the transcription factors STAT3 and c-Jun had only minor roles in IFN-γ induced expression of PD-L1. The mechanism underlying constitutive expression remains unresolved. We suggest these results have significance in selection of treatments that can be used in combination with monoclonal antibodies against PD1, to enhance their effectiveness and to reduce inhibitory effects melanoma cells have against cytotoxic T cell activity.

  14. Constitutive expression of Wnt/β-catenin target genes promotes proliferation and invasion of liver cancer stem cells

    Science.gov (United States)

    CHEN, WEI; ZHANG, YU-WEI; LI, YANG; ZHANG, JIAN-WEN; ZHANG, TONG; FU, BIN-SHENG; ZHANG, QI; JIANG, NAN

    2016-01-01

    Wnt/β-catenin is an important signaling pathways involved in the tumorgenesis, progression and maintenance of cancer stem cells (CSCs). In the present study, the role of Wnt/β-catenin signaling in CSC-mediated tumorigenesis and invasion in liver CSCs was investigated. A small population of cancer stem-like side population (SP) cells (3.6%) from liver cancer samples were identified. The cells were highly resistant to drug treatment due to the enhanced expression of drug efflux pumps, such as ABC subfamily G member 2, multidrug resistance protein 1 and ATP-binding cassette subfamily B member 5. Furthermore, using TOPflash and reverse transcription-quantitative polymerase chain reaction analysis, Wnt/β-catenin signaling and the transcriptional regulation of Wnt/β-catenin target genes including dickkopf Wnt signaling pathway inhibitor 1, axis inhibition protein 2 and cyclin D1 were observed to be markedly upregulated in liver cancer SP cells. As a consequence, SP cells possessed infinite cell proliferation potential and the ability to generating tumor spheres. In addition, upon reducing Wnt/β-catenin signaling, the rates of proliferation, tumor sphere formation and tumor invasion of SP cells were markedly reduced. Therefore, these data suggest that Wnt/β-catenin signaling is a potential therapeutic target to reduce CSC-mediated tumorigenicity and invasion in liver cancer. PMID:26956539

  15. The Physiological and Biochemical Mechanisms Providing the Increased Constitutive Cold Resistance in the Potato Plants, Expressing the Yeast SUC2 Gene Encoding Apoplastic Invertase

    Directory of Open Access Journals (Sweden)

    A.N. Deryabin

    2016-05-01

    Full Text Available The expression of heterologous genes in plants is an effective method to improve our understanding of plant resistance mechanisms. The purpose of this work was to investigate the involvement of cell-wall invertase and apoplastic sugars into constitutive cold resistance of potato (Solanum tuberosum L., cv. Dйsirйe plants, which expressed the yeast SUC2 gene encoding apoplastic invertase. WT-plants of a potato served as the control. The increase in the essential cell-wall invertase activity in the leaves of transformed plants indicates significant changes in the cellular carbohydrate metabolism and regulatory function of this enzyme. The activity of yeast invertase changed the composition of intracellular sugars in the leaves of the transformed potato plant. The total content of sugars (sucrose, glucose, fructose in the leaves and apoplast was higher in the transformants, in comparison by WT-plants. Our data indicate higher constitutive resistance of transformants to severe hypothermia conditions compared to WT-plants. This fact allows us to consider cell-wall invertase as a enzyme of carbohydrate metabolism playing an important regulatory role in the metabolic signaling upon forming increased plant resistance to low temperature. Thus, the potato line with the integrated SUC2 gene is a convenient tool to study the role of the apoplastic invertase and the products of its activity during growth, development and formation constitutive resistance to hypothermia.

  16. Constitutive knockout of Surf1 is associated with high embryonic lethality, mitochondrial disease and cytochrome c oxidase deficiency in mice.

    Science.gov (United States)

    Agostino, Alessandro; Invernizzi, Federica; Tiveron, Cecilia; Fagiolari, Gigliola; Prelle, Alessandro; Lamantea, Eleonora; Giavazzi, Alessio; Battaglia, Giorgio; Tatangelo, Laura; Tiranti, Valeria; Zeviani, Massimo

    2003-02-15

    We report here the creation of a constitutive knockout mouse for SURF1, a gene encoding one of the assembly proteins involved in the formation of cytochrome c oxidase (COX). Loss-of-function mutations of SURF1 cause Leigh syndrome associated with an isolated and generalized COX deficiency in humans. The murine phenotype is characterized by the following hallmarks: (1) high post-implantation embryonic lethality, affecting approximately 90% of the Surf1(-/-) individuals; (2) early-onset mortality of post-natal individuals; (3) highly significant deficit in muscle strength and motor performance; (4) profound and isolated defect of COX activity in skeletal muscle and liver, and, to a lesser extent, heart and brain; (5) morphological abnormalities of skeletal muscle, characterized by reduced histochemical reaction to COX and mitochondrial proliferation; (6) no obvious abnormalities in brain morphology, reflecting the virtual absence of overt neurological symptoms. These results indicate a function for murine Surf1 protein (Surf1p) specifically related to COX and recapitulate, at least in part, the human phenotype. This is the first mammalian model for a nuclear disease gene of a human mitochondrial disorder. Our model constitutes a useful tool to investigate the function of Surf1p, help understand the pathogenesis of Surf1p deficiency in vivo, and evaluate the efficacy of treatment.

  17. Constitutive expression of MC1R in HaCaT keratinocytes inhibits basal and UVB-induced TNF-alpha production.

    Science.gov (United States)

    Garcin, Geneviève; Le Gallic, Lionel; Stoebner, Pierre-Emmanuel; Guezennec, Anne; Guesnet, Joelle; Lavabre-Bertrand, Thierry; Martinez, Jean; Meunier, Laurent

    2009-01-01

    Alpha-melanocyte stimulating hormone (alpha-MSH) binds to melanocortin-1 receptor (MC1R) on melanocytes to stimulate pigmentation and modulate various cutaneous inflammatory responses. MC1R expression is not restricted to melanocytic cells and may be induced in keratinocytes after UVB exposure. We hypothesized that MC1R signaling in keratinocytes, wherein basal conditions are barely expressed, may modulate mediators of inflammation, such as nuclear factor-kappa B (NF-kappaB) and tumor necrosis factor-alpha (TNF-alpha). Therefore, we generated HaCaT cells that stably express human MC1R or the Arg151Cys (R151C) nonfunctional variant. We demonstrate that: (1) the constitutive activity of MC1R results in elevated intracellular cAMP level, reduced NF-kappaB activity and decreased TNF-alpha transcription; (2) binding of alpha-MSH to MC1R and the subsequent increase in cAMP production do not inhibit TNFalpha-mediated NF-kappaB activation; (3) MC1R signaling is sufficient to strongly inhibit UVB-induced TNF-alpha expression and this inhibitory effect is further enhanced by alpha-MSH stimulation. Our findings suggest that the constitutive activity of the G-protein-coupled MC1R in keratinocytes may contribute to the modulation of inflammatory events and immune response induced by UV light.

  18. Constitutive equations of basalt filament tows under quasi-static and high strain rate tension

    Energy Technology Data Exchange (ETDEWEB)

    Zhu Lvtao; Sun Baozhong [College of Textiles, Donghua University, Shanghai 201620 (China); Hu, Hong [Institute of Textiles and Clothing, Hong Kong Polytechnic University, Hung Hom Kowloon (Hong Kong); Gu Bohong, E-mail: gubh@dhu.edu.cn [College of Textiles, Donghua University, Shanghai 201620 (China); Department of Textile Engineering, Zhongyuan Institute of Technology, Zhengzhou, Henan Province 450007 (China)

    2010-05-25

    The tensile properties of basalt filament tows were tested at quasi-static (0.001 s{sup -1}) and high strain rates (up to 3000 s{sup -1}) with MTS materials tester (MTS 810.23) and split Hopkinson tension bar (SHTB), respectively. Experimental results showed that the mechanical properties of the basalt filament tows were rather sensitive to strain rate. Specifically, the stiffness and failure stress of the basalt filament tows increased distinctly as the strain rate increased, while the failure strain decreased. From scanning electronic microscope (SEM) photographs of the fracture surface, it is indicated that the basalt filament tows failed in a more brittle mode and the fracture surface got more regular as the strain rate increases. The strength distributions of the basalt filament tows have been evaluated by a single Weibull distribution function. The curve predicted from the single Weibull distribution function was in good agreement with the experimental data points.

  19. High-Temperature Deformation Constitutive Law for Dissimilar Weld Residual Stress Modeling: Effect of Thermal Load on Strain Hardening

    Energy Technology Data Exchange (ETDEWEB)

    Yu, Xinghua [ORNL; Wang, Yanli [ORNL; Crooker, Paul [Electric Power Research Institute (EPRI); Feng, Zhili [ORNL

    2015-01-01

    Weld residual stress is one of the primary driving forces for primary water stress corrosion cracking in dissimilar metal welds (DMWs). To mitigate tensile residual stress in DMWs, it is critical to understand residual stress distribution by modeling techniques. Recent studies have shown that weld residual stress prediction using today s DMW residual stress models strongly depends on the strain-hardening constitutive model chosen. The commonly used strain-hardening models (isotropic, kinematic, and mixed) are all time-independent and inadequate to account for the time-dependent (viscous) plastic deformation at the elevated temperatures experienced during welding. For materials with profound strain-hardening, such as stainless steels and nickel-based alloys that are widely used in nuclear reactor and piping systems, the equivalent plastic strain the determinate factor of the flow stress can be highly dependent on the recovery and recrystallization processes. These processes are in turn a strong function of temperature, time, and deformation rate. Recently, the authors proposed a new temperature- and time-dependent strain-hardening constitutive model: the dynamic strain-hardening constitutive model. The application of such a model has resulted in improved weld residual stress prediction compared to the residual stress measurement results from the contour and deep-hole drilling methods. In this study, the dynamic strain-hardening behavior of Type 304 stainless steel and Alloy 82 used in pressure vessel nozzle DMWs is experimentally determined. The kinetics of the recovery and recrystallization of flow stress are derived from experiments, resulting in a semi-empirical equation as a function of pre-strain, time, and temperature that can be used for weld residual stress modeling. The method used in this work also provides an approach to study the kinetics of recovery and recrystallization of other materials with significant strain-hardening.

  20. The in vitro and in vivo effects of constitutive light expression on a bioluminescent strain of the mouse enteropathogen Citrobacter rodentium

    Directory of Open Access Journals (Sweden)

    Hannah M. Read

    2016-06-01

    Full Text Available Bioluminescent reporter genes, such as those from fireflies and bacteria, let researchers use light production as a non-invasive and non-destructive surrogate measure of microbial numbers in a wide variety of environments. As bioluminescence needs microbial metabolites, tagging microorganisms with luciferases means only live metabolically active cells are detected. Despite the wide use of bioluminescent reporter genes, very little is known about the impact of continuous (also called constitutive light expression on tagged bacteria. We have previously made a bioluminescent strain of Citrobacter rodentium, a bacterium which infects laboratory mice in a similar way to how enteropathogenic Escherichia coli (EPEC and enterohaemorrhagic E. coli (EHEC infect humans. In this study, we compared the growth of the bioluminescent C. rodentium strain ICC180 with its non-bioluminescent parent (strain ICC169 in a wide variety of environments. To understand more about the metabolic burden of expressing light, we also compared the growth profiles of the two strains under approximately 2,000 different conditions. We found that constitutive light expression in ICC180 was near-neutral in almost every non-toxic environment tested. However, we also found that the non-bioluminescent parent strain has a competitive advantage over ICC180 during infection of adult mice, although this was not enough for ICC180 to be completely outcompeted. In conclusion, our data suggest that constitutive light expression is not metabolically costly to C. rodentium and supports the view that bioluminescent versions of microbes can be used as a substitute for their non-bioluminescent parents to study bacterial behaviour in a wide variety of environments.

  1. Constitutive modeling of creep behavior in single crystal superalloys: Effects of rafting at high temperatures

    Energy Technology Data Exchange (ETDEWEB)

    Fan, Ya-Nan, E-mail: fanyn12@mails.tsinghua.edu.cn; Shi, Hui-Ji, E-mail: shihj@mail.tsinghua.edu.cn; Qiu, Wen-Hui

    2015-09-17

    Rafting and creep modeling of single crystal superalloys at high temperatures are important for the safety assessment and life prediction in practice. In this research, a new model has been developed to describe the rafting evolution and incorporated into the Cailletaud single crystal plasticity model to simulate the creep behavior. The driving force of rafting is assumed to be the relaxation of the strain energy, and it is calculated with the local stress state, a superposition of the external and misfit stress tensors. In addition, the isotropic coarsening is introduced by the cube root dependence of the microstructure periodicity on creep time based on Ostwal ripening. Then the influence of rafting on creep deformation is taken into account as the Orowan stress in the single crystal plasticity model. The capability of the proposed model is validated with creep experiments of CMSX-4 at 950 °C and 1050 °C. It is able to predict the rafting direction at complex loading conditions and evaluate the channel width during rafting. For [001] tensile creep tests, good agreement has been shown between the model predictions and experimental results at different temperatures and stress levels. The creep acceleration can be captured with this model and is attributed to the microstructure degradation caused by the precipitate coarsening.

  2. On the nature of facultative and constitutive CAM: environmental and developmental control of CAM expression during early growth of Clusia, Kalanchöe, and Opuntia.

    Science.gov (United States)

    Winter, Klaus; Garcia, Milton; Holtum, Joseph A M

    2008-01-01

    The capacity to induce crassulacean acid metabolism developmentally (constitutive CAM) and to up-regulate CAM expression in response to drought stress (facultative CAM) was studied in whole shoots of seven species by measuring net CO(2) gas exchange for up to 120 day-night cycles during early growth. In Clusia rosea, CAM was largely induced developmentally. Well-watered seedlings began their life cycle as C(3) plants and developed net dark CO(2) fixation indicative of CAM after the initiation of the fourth leaf pair following the cotyledons. Thereafter, CAM activity increased progressively and drought stress led to only small additional, reversible increases in dark CO(2) fixation. In contrast, CAM expression was overwhelmingly under environmental control in seedlings and mature plants of Clusia pratensis. C(3)-type CO(2) exchange was maintained under well-watered conditions, but upon drought stress, CO(2) exchange shifted, in a fully reversible manner, to a CAM-type pattern. Clusia minor showed CO(2) exchange reponses intermediate to those of C. rosea and C. pratensis. Clusia cretosa operated in the C(3) mode at all times. Notably, reversible stress-induced increases of dark CO(2) fixation were also observed during the developmental progression to pronounced CAM in young Kalanchoë daigremontiana and Kalanchoë pinnata, two species considered constitutive CAM species. Drought-induced up-regulation of CAM was even detected in young cladodes of a cactus, Opuntia ficus-indica, an archetypal constitutive CAM species. Evidently, the defining characteristics of constitutive and facultative CAM are shared, to variable degrees, by all CAM species.

  3. Constitutive expression and enzymatic activity of Tan protein in brain and epidermis of Ceratitis capitata and of Drosophila melanogaster wild-type and tan mutants.

    Science.gov (United States)

    Pérez, M M; Sabio, G; Badaracco, A; Quesada-Allué, L A

    2011-09-01

    The present report shows a partial biochemical characterization and life cycle expression of N-β-alanyldopamine hydrolase (Tan protein) in Ceratitis capitata and Drosophila melanogaster. This enzyme catalyzes the hydrolysis of N-β-alanyldopamine (NBAD), the main tanning precursor of insect brown cuticles. It also plays an important role in the metabolism of brain neurotransmitters, recycling dopamine and histamine. In contrast to NBAD-synthase, Tan is expressed constitutively in epidermis and does not respond directly to microbial challenge. Immunodetection experiments showed the novel localization of NBAD-hydrolase in the embryo central neural system and in different regions of the adult brain, in addition to optic lobes. We sequenced and characterized Drosophila mutants tan¹ and tan³. The latter appears to be a mutant with normal expression in neural tissue but weak one in epidermis. Copyright © 2011 Elsevier Ltd. All rights reserved.

  4. Prostacyclin Synthase: Upregulation during Renal Development and in Glomerular Disease as well as Its Constitutive Expression in Cultured Human Mesangial Cells

    Directory of Open Access Journals (Sweden)

    Thomas Klein

    2015-01-01

    Full Text Available Prostacyclin (PGI2 plays a critical role in nephrogenesis and renal physiology. However, our understanding of how prostacyclin release in the kidney is regulated remains poorly defined. We studied expression of prostacyclin synthase (PGIS in developing and adult human kidneys, and also in selected pediatric renal diseases. We also examined PGI2 formation in human mesangial cells in vitro. We observed abundant expression of PGIS in the nephrogenic cortex in humans and in situ hybridization revealed an identical pattern in mice. In the normal adult kidney, PGIS-immunoreactive protein and mRNA appear to localize to mesangial fields and endothelial and smooth muscle cells of arteries and peritubular capillaries. In kidney biopsies taken from pediatric patients, enhanced expression of PGIS-immunoreactive protein was noted mainly in endothelial cells of patients with IgA-nephropathy. Cultured human mesangial cells produce primarily PGI2 and prostaglandin E2, followed by prostaglandin F2α Cytokine stimulation increased PGI2 formation 24-fold. Under these conditions expression of PGIS mRNA and protein remained unaltered whereas mRNA for cyclooxygenase-2 was markedly induced. In contrast to its constitutive expression in vitro, renal expression of prostacyclin-synthase appears to be regulated both during development and in glomerular disease. Further research is needed to identify the factors involved in regulation of PGIS-expression.

  5. The active form of goat insulin-like peptide 3 (INSL3) is a single-chain structure comprising three domains B-C-A, constitutively expressed and secreted by testicular Leydig cells.

    Science.gov (United States)

    Siqin; Minagawa, Itaru; Okuno, Mitsutoshi; Yamada, Kimihiko; Sugawara, Yasushi; Nagura, Yoshio; Hamano, Koh-Ichi; Park, Enoch Y; Sasada, Hiroshi; Kohsaka, Tetsuya

    2013-09-01

    Relaxin-like factor (RLF), also called insulin-like peptide 3 (INSL3), is a member of the insulin/relaxin gene family and is produced by testicular Leydig cells. While the understanding of its effects is growing, very little is known about the structural and functional properties of native INSL3. Here, we demonstrate that native INSL3 isolated from goat testes is a single-chain structure with full biological activity, and is constitutively expressed and secreted by Leydig cells. Using a series of chromatography steps, native INSL3 was highly purified as a single 12-kDa peak as revealed by SDS-PAGE. MS/MS analysis provided 81% sequence coverage and revealed a distinct single-chain structure consisting of the B-, C-, and A-domains deduced previously from the INSL3 cDNA sequence. Moreover, the N-terminal peptide was six amino acid residues longer than predicted. Native INSL3 exhibited full bioactivity in HEK-293 cells expressing the receptor for INSL3. Immunoelectron microscopy and Western blot analysis revealed that INSL3 was secreted by Leydig cells through the constitutive pathway into blood and body fluids. We conclude, therefore, that goat INSL3 is constitutively secreted from Leydig cells as a B-C-A single-chain structure with full biological activity.

  6. An OMV Vaccine Derived from a Capsular Group B Meningococcus with Constitutive FetA Expression: Preclinical Evaluation of Immunogenicity and Toxicity.

    Directory of Open Access Journals (Sweden)

    Gunnstein Norheim

    Full Text Available Following the introduction of effective protein-polysaccharide conjugate vaccines against capsular group C meningococcal disease in Europe, meningococci of capsular group B remain a major cause of death and can result in debilitating sequelae. The outer membrane proteins PorA and FetA have previously been shown to induce bactericidal antibodies in humans. Despite considerable antigenic variation among PorA and FetA OMPs in meningococci, systematic molecular epidemiological studies revealed this variation is highly structured so that a limited repertoire of antigenic types is congruent with the hyperinvasive meningococcal lineages that have caused most of the meningococcal disease in Europe in recent decades. Here we describe the development of a prototype vaccine against capsular group B meningococcal infection based on a N. meningitidis isolate genetically engineered to have constitutive expression of the outer membrane protein FetA. Deoxycholate outer membrane vesicles (dOMVs extracted from cells cultivated in modified Frantz medium contained 21.8% PorA protein, 7.7% FetA protein and 0.03 μg LPS per μg protein (3%. The antibody response to the vaccine was tested in three mouse strains and the toxicological profile of the vaccine was tested in New Zealand white rabbits. Administration of the vaccine, MenPF-1, when given by intramuscular injection on 4 occasions over a 9 week period, was well tolerated in rabbits up to 50 μg/dose, with no evidence of systemic toxicity. These data indicated that the MenPF-1 vaccine had a toxicological profile suitable for testing in a phase I clinical trial.

  7. An OMV Vaccine Derived from a Capsular Group B Meningococcus with Constitutive FetA Expression: Preclinical Evaluation of Immunogenicity and Toxicity.

    Science.gov (United States)

    Norheim, Gunnstein; Sanders, Holly; Mellesdal, Jardar W; Sundfør, Idunn; Chan, Hannah; Brehony, Carina; Vipond, Caroline; Dold, Chris; Care, Rory; Saleem, Muhammad; Maiden, Martin C J; Derrick, Jeremy P; Feavers, Ian; Pollard, Andrew J

    2015-01-01

    Following the introduction of effective protein-polysaccharide conjugate vaccines against capsular group C meningococcal disease in Europe, meningococci of capsular group B remain a major cause of death and can result in debilitating sequelae. The outer membrane proteins PorA and FetA have previously been shown to induce bactericidal antibodies in humans. Despite considerable antigenic variation among PorA and FetA OMPs in meningococci, systematic molecular epidemiological studies revealed this variation is highly structured so that a limited repertoire of antigenic types is congruent with the hyperinvasive meningococcal lineages that have caused most of the meningococcal disease in Europe in recent decades. Here we describe the development of a prototype vaccine against capsular group B meningococcal infection based on a N. meningitidis isolate genetically engineered to have constitutive expression of the outer membrane protein FetA. Deoxycholate outer membrane vesicles (dOMVs) extracted from cells cultivated in modified Frantz medium contained 21.8% PorA protein, 7.7% FetA protein and 0.03 μg LPS per μg protein (3%). The antibody response to the vaccine was tested in three mouse strains and the toxicological profile of the vaccine was tested in New Zealand white rabbits. Administration of the vaccine, MenPF-1, when given by intramuscular injection on 4 occasions over a 9 week period, was well tolerated in rabbits up to 50 μg/dose, with no evidence of systemic toxicity. These data indicated that the MenPF-1 vaccine had a toxicological profile suitable for testing in a phase I clinical trial.

  8. Conserved but attenuated parental gene expression in allopolyploids: constitutive zinc hyperaccumulation in the allotetraploidArabidopsis kamchatica

    OpenAIRE

    Paape, Timothy; Hatakeyama, Masaomi; Shimizu-Inatsugi, Rie; Cereghetti, Teo; Onda, Yoshihiko; Kenta, Tanaka; Sese, Jun; Kentaro K. Shimizu

    2016-01-01

    Allopolyploidization combines parental genomes and often confers broader species distribution. However, little is known about parentally transmitted gene expression underlying quantitative traits following allopolyploidization because of the complexity of polyploid genomes. The allopolyploid species Arabidopsis kamchatica is a natural hybrid of the zinc hyperaccumulator Arabidopsis halleri and of the nonaccumulator Arabidopsis lyrata. We found that A. kamchatica retained the ability to hypera...

  9. Symbolic derivation of potential based constitutive equations

    Science.gov (United States)

    Arnold, S. M.; Tan, H. Q.

    1990-05-01

    Structural alloys used in high temperature applications exhibit complex thermomechanical behavior that is inherently time dependent and hereditary, as the current behavior depends not only on current conditions but on the thermomechanical history. Derivation of mathematical expressions (constitutive equations) which describe this high temperature material behavior can be quite time consuming, involved, and error-prone, thus intelligent application of symbolic systems to facilitate this tedious processes can be of significant benefit. Here a computerized package, running under MACSYMA, capable of efficiently deriving potential based constitutive models, in analytical form (involving tensors, partial differentiation, invariants, and the like) is presented. Special purpose utility algorithms are designed and implemented to perform partial differentiation (chain rule), tensor manipulation, case distinction and simplification. Four constitutive theories reported in the literature are utilized to verify implementation accuracy. It is expected that this symbolic package can and will provide a significant incentive to the development of new constitutive theories.

  10. Constitutive expression of the xylanase inhibitor TAXI-III delays Fusarium head blight symptoms in durum wheat transgenic plants.

    Science.gov (United States)

    Moscetti, Ilaria; Tundo, Silvio; Janni, Michela; Sella, Luca; Gazzetti, Katia; Tauzin, Alexandra; Giardina, Thierry; Masci, Stefania; Favaron, Francesco; D'Ovidio, Renato

    2013-12-01

    Cereals contain xylanase inhibitor (XI) proteins which inhibit microbial xylanases and are considered part of the defense mechanisms to counteract microbial pathogens. Nevertheless, in planta evidence for this role has not been reported yet. Therefore, we produced a number of transgenic plants constitutively overexpressing TAXI-III, a member of the TAXI type XI that is induced by pathogen infection. Results showed that TAXI-III endows the transgenic wheat with new inhibition capacities. We also showed that TAXI-III is correctly secreted into the apoplast and possesses the expected inhibition parameters against microbial xylanases. The new inhibition properties of the transgenic plants correlate with a significant delay of Fusarium head blight disease symptoms caused by Fusarium graminearum but do not significantly influence leaf spot symptoms caused by Bipolaris sorokiniana. We showed that this contrasting result can be due to the different capacity of TAXI-III to inhibit the xylanase activity of these two fungal pathogens. These results provide, for the first time, clear evidence in planta that XI are involved in plant defense against fungal pathogens and show the potential to manipulate TAXI-III accumulation to improve wheat resistance against F. graminearum.

  11. Transposon-mediated alteration of TaMATE1B expression in wheat confers constitutive citrate efflux from root apices.

    Science.gov (United States)

    Tovkach, Andriy; Ryan, Peter R; Richardson, Alan E; Lewis, David C; Rathjen, Tina M; Ramesh, Sunita; Tyerman, Stephen D; Delhaize, Emmanuel

    2013-02-01

    The TaMATE1B gene (for multidrug and toxic compound extrusion) from wheat (Triticum aestivum) was isolated and shown to encode a citrate transporter that is located on the plasma membrane. TaMATE1B expression in roots was induced by iron deficiency but not by phosphorus deficiency or aluminum treatment. The coding region of TaMATE1B was identical in a genotype showing citrate efflux from root apices (cv Carazinho) to one that lacked citrate efflux (cv Egret). However, sequence upstream of the coding region differed between these two genotypes in two ways. The first difference was a single-nucleotide polymorphism located approximately 2 kb upstream from the start codon in cv Egret. The second difference was an 11.1-kb transposon-like element located 25 bp upstream of the start codon in cv Carazinho that was absent from cv Egret. The influence of these polymorphisms on TaMATE1B expression was investigated using fusions to green fluorescent protein expressed in transgenic lines of rice (Oryza sativa). Fluorescence measurements in roots of rice indicated that 1.5- and 2.3-kb regions upstream of TaMATE1B in cv Carazinho (which incorporated 3' regions of the transposon-like element) generated 20-fold greater expression in the apical 1 mm of root compared with the native promoter in cv Egret. By contrast, fluorescence in more mature tissues was similar in both cultivars. The presence of the single-nucleotide polymorphism alone consistently generated 2-fold greater fluorescence than the cv Egret promoter. We conclude that the transposon-like element in cv Carazinho extends TaMATE1B expression to the root apex, where it confers citrate efflux and enhanced aluminum tolerance.

  12. The constitutive expression of the V gene of Parainfluenza virus 5 affects the growth properties of bovine herpesvirus 5

    Directory of Open Access Journals (Sweden)

    Francisco Esmaile de Sales Lima

    2014-02-01

    Full Text Available This study aimed to analyze the effect of the expression of Parainfluenza virus 5 (PIV5 V protein in bovine cells on the replication of Bovine herpesvirus 5 (BoHV-5. Growth properties of BoHV-5 were evaluated in parental and PIV5 transfected cells. In one-step growth experiments, the BoHV-5 reached higher titers at earlier time points in the transfected cells when compared to the parental cells. The mean plaque size produced by the BoHV-5 in transfected cells was larger than the parental cells. This indicated that the expression of the PIV5 V gene facilitated the release and cell-to-cell spread of BoHV-5 in bovine cells.

  13. Constitutive Expression of OsIAA9 Affects Starch Granules Accumulation and Root Gravitropic Response in Arabidopsis.

    Science.gov (United States)

    Luo, Sha; Li, Qianqian; Liu, Shanda; Pinas, Nicholaas M; Tian, Hainan; Wang, Shucai

    2015-01-01

    Auxin/Indole-3-Acetic Acid (Aux/IAA) genes are early auxin response genes ecoding short-lived transcriptional repressors, which regulate auxin signaling in plants by interplay with Auxin Response Factors (ARFs). Most of the Aux/IAA proteins contain four different domains, namely Domain I, Domain II, Domain III, and Domain IV. So far all Aux/IAA mutants with auxin-related phenotypes identified in both Arabidopsis and rice (Oryza sativa) are dominant gain-of-function mutants with mutations in Domain II of the corresponding Aux/IAA proteins, suggest that Aux/IAA proteins in both Arabidopsis and rice are largely functional redundantly, and they may have conserved functions. We report here the functional characterization of a rice Aux/IAA gene, OsIAA9. RT-PCR results showed that expression of OsIAA9 was induced by exogenously applied auxin, suggesting that OsIAA9 is an auxin response gene. Bioinformatic analysis showed that OsIAA9 has a repressor motif in Domain I, a degron in Domain II, and the conserved amino acid signatures for protein-protein interactions in Domain III and Domain IV. By generating transgenic plants expressing GFP-OsIAA9 and examining florescence in the transgenic plants, we found that OsIAA9 is localized in the nucleus. When transfected into protoplasts isolated from rosette leaves of Arabidopsis, OsIAA9 repressed reporter gene expression, and the repression was partially released by exogenously IAA. These results suggest that OsIAA9 is a canonical Aux/IAA protein. Protoplast transfection assays showed that OsIAA9 interacted ARF5, but not ARF6, 7, 8 and 19. Transgenic Arabidopsis plants expressing OsIAA9 have increased number of lateral roots, and reduced gravitropic response. Further analysis showed that OsIAA9 transgenic Arabidopsis plants accumulated fewer granules in their root tips and the distribution of granules was also affected. Taken together, our study showed that OsIAA9 is a transcriptional repressor, and it regulates gravitropic

  14. Constitutive expression of OsIAA9 affects starch granules accumulation and root gravitropic response in Arabidopsis

    Directory of Open Access Journals (Sweden)

    Sha eLuo

    2015-12-01

    Full Text Available Auxin/Indole-3-Acetic Acid (Aux/IAA genes are early auxin response genes ecoding short-lived transcriptional repressors, which regulate auxin signaling in plants by interplay with Auxin Response Factors (ARFs. Most of the Aux/IAA proteins contain four different domains, namely Domain I, Domain II, Domain III and Domain IV. So far all Aux/IAA mutants with auxin-related phenotypes identified in both Arabidopsis and rice (Oryza sativa are dominant gain-of-function mutants with mutations in Domain II of the corresponding Aux/IAA proteins, suggest that Aux/IAA proteins in both Arabidopsis and rice are largely functional redundantly, and they may have conserved functions. We report here the functional characterization of a rice Aux/IAA gene, OsIAA9. RT-PCR results showed that expression of OsIAA9 was induced by exogenously applied auxin, suggesting that OsIAA9 is an auxin response gene. Bioinformatic analysis showed that OsIAA9 has a repressor motif in Domain I, a degron in Domain II, and the conserved amino acid signatures for protein-protein interactions in Domain III and Domain IV. By generating transgenic plants expressing GFP-OsIAA9 and examining florescence in the transgenic plants, we found that OsIAA9 is localized in the nucleus. When transfected into protoplasts isolated from rosette leaves of Arabidopsis, OsIAA9 repressed reporter gene expression, and the repression was partially released by exogenously IAA. These results suggest that OsIAA9 is a canonical Aux/IAA protein. Protoplast transfection assays showed that OsIAA9 interacted ARF5, but not ARF6, 7, 8 and 19. Transgenic Arabidopsis plants expressing OsIAA9 have increased number of lateral roots, and reduced gravitropic response. Further analysis showed that OsIAA9 transgenic Arabidopsis plants accumulated fewer granules in their root tips and the distribution of granules was also affected. Taken together, our study showed that OsIAA9 is a transcriptional repressor, and it regulates

  15. ALK1 signalling analysis identifies angiogenesis related genes and reveals disparity between TGF-β and constitutively active receptor induced gene expression

    Directory of Open Access Journals (Sweden)

    Hafner Mathias

    2006-04-01

    Full Text Available Abstract Background TGF-β1 is an important angiogenic factor involved in the different aspects of angiogenesis and vessel maintenance. TGF-β signalling is mediated by the TβRII/ALK5 receptor complex activating the Smad2/Smad3 pathway. In endothelial cells TGF-β utilizes a second type I receptor, ALK1, activating the Smad1/Smad5 pathway. Consequently, a perturbance of ALK1, ALK5 or TβRII activity leads to vascular defects. Mutations in ALK1 cause the vascular disorder hereditary hemorrhagic telangiectasia (HHT. Methods The identification of ALK1 and not ALK5 regulated genes in endothelial cells, might help to better understand the development of HHT. Therefore, the human microvascular endothelial cell line HMEC-1 was infected with a recombinant constitutively active ALK1 adenovirus, and gene expression was studied by using gene arrays and quantitative real-time PCR analysis. Results After 24 hours, 34 genes were identified to be up-regulated by ALK1 signalling. Analysing ALK1 regulated gene expression after 4 hours revealed 13 genes to be up- and 2 to be down-regulated. Several of these genes, including IL-8, ET-1, ID1, HPTPη and TEAD4 are reported to be involved in angiogenesis. Evaluation of ALK1 regulated gene expression in different human endothelial cell types was not in complete agreement. Further on, disparity between constitutively active ALK1 and TGF-β1 induced gene expression in HMEC-1 cells and primary HUVECs was observed. Conclusion Gene array analysis identified 49 genes to be regulated by ALK1 signalling and at least 14 genes are reported to be involved in angiogenesis. There was substantial agreement between the gene array and quantitative real-time PCR data. The angiogenesis related genes might be potential HHT modifier genes. In addition, the results suggest endothelial cell type specific ALK1 and TGF-β signalling.

  16. High Cell Density Process for Constitutive Production of a Recombinant Phytase in Thermotolerant Methylotrophic Yeast Ogataea thermomethanolica Using Table Sugar as Carbon Source.

    Science.gov (United States)

    Charoenrat, Theppanya; Antimanon, Sompot; Kocharin, Kanokarn; Tanapongpipat, Sutipa; Roongsawang, Niran

    2016-12-01

    The yeast Ogataea thermomethanolica has recently emerged as a potential host for heterologous protein expression at elevated temperature. To evaluate the feasibility of O. thermomethanolica as heterologous host in large-scale fermentation, constitutive production of fungal phytase was investigated in fed-batch fermentation. The effect of different temperatures, substrate feeding strategies, and carbon sources on phytase production was investigated. It was found that O. thermomethanolica can grow in the temperature up to 40 °C and optimal at 34 °C. However, the maximum phytase production was observed at 30 °C and slightly decreased at 34 °C. The DOT stat control was the most efficient feeding strategy to obtain high cell density and avoid by-product formation. The table sugar can be used as an alternative substrate for phytase production in O. thermomethanolica. The highest phytase activity (134 U/mL) was obtained from table sugar at 34 °C which was 20-fold higher than batch culture (5.7 U/mL). At a higher cultivation temperature of 38 °C, table sugar can be used as a low-cost substrate for the production of phytase which was expressed with an acceptable yield (85 U/mL). Lastly, the results from this study reveal the industrial favorable benefits of employing O. thermomethanolica as a host for heterologous protein production.

  17. Generation of a constitutively expressing Tetracycline repressor (TetR human embryonic stem cell line BJNhem20-TetR

    Directory of Open Access Journals (Sweden)

    Ronak Shetty

    2016-03-01

    Full Text Available Human embryonic stem cell line BJNhem20-TetR was generated using non-viral method. The construct pCAG-TetRnls was transfected using microporation procedure. BJNhem20-TetR can subsequently be transfected with any vector harbouring a TetO (Tet operator sequence to generate doxycycline based inducible line. For example, in human embryonic stem cells, the pSuperior based TetO system has been transfected into a TetR containing line to generate OCT4 knockdown cell line (Zafarana et al., 2009. Thus BJNhem20-TetR can be used as a tool to perturb gene expression in human embryonic stem cells.

  18. Increased Na⁺/Ca²⁺ exchanger expression/activity constitutes a point of inflection in the progression to heart failure of hypertensive rats.

    Directory of Open Access Journals (Sweden)

    Jesica S Rodriguez

    Full Text Available Spontaneously hypertensive rat (SHR constitutes a genetic model widely used to study the natural evolution of hypertensive heart disease. Ca²⁺-handling alterations are known to occur in SHR. However, the putative modifications of Ca²⁺-handling proteins during the progression to heart failure (HF are not well established. Moreover, the role of apoptosis in SHR is controversial. We investigated intracellular Ca²⁺, Ca²⁺-handling proteins and apoptosis in SHR vs. control Wistar rats (W from 3 to 15 months (mo. Changes associated with the transition to HF (i.e. lung edema and decrease in midwall fractional shortening, occurred at 15 mo in 38% of SHR (SHRF. In SHRF, twitch and caffeine-induced Ca²⁺ transients, significantly decreased relative to 6/9 mo and 15 mo without HF signs. This decrease occurred in association with a decrease in the time constant of caffeine-Ca²⁺ transient decay and an increase in Na⁺/Ca²⁺ exchanger (NCX abundance (p<0.05 with no changes in SERCA2a expression/activity. An increased Ca²⁺-calmodulin-kinase II activity, associated with an enhancement of apoptosis (TUNEL and Bax/Bcl2 was observed in SHR relative to W from 3 to 15 mo.1. Apoptosis is an early and persistent event that may contribute to hypertrophic remodeling but would not participate in the contractile impairment of SHRF. 2. The increase in NCX expression/activity, associated with an increase in Ca²⁺ efflux from the cell, constitutes a primary alteration of Ca²⁺-handling proteins in the evolution to HF. 3. No changes in SERCA2a expression/activity are observed when HF signs become evident.

  19. Constitutive expression of the tzs gene from Agrobacterium tumefaciens virG mutant strains is responsible for improved transgenic plant regeneration in cotton meristem transformation.

    Science.gov (United States)

    Ye, Xudong; Chen, Yurong; Wan, Yuechun; Hong, Yun-Jeong; Ruebelt, Martin C; Gilbertson, Larry A

    2016-03-01

    KEY MESSAGE : virG mutant strains of a nopaline type of Agrobacterium tumefaciens increase the transformation frequency in cotton meristem transformation. Constitutive cytokinin expression from the tzs gene in the virG mutant strains is responsible for the improvement. Strains of Agrobacterium tumefaciens were tested for their ability to improve cotton meristem transformation frequency. Two disarmed A. tumefaciens nopaline strains with either a virGN54D constitutively active mutation or virGI77V hypersensitive induction mutation significantly increased the transformation frequency in a cotton meristem transformation system. The virG mutant strains resulted in greener explants after three days of co-culture in the presence of light, which could be attributed to a cytokinin effect of the mutants. A tzs knockout strain of virGI77V mutant showed more elongated, less green explants and decreased cotton transformation frequency, as compared to a wild type parental strain, suggesting that expression of the tzs gene is required for transformation frequency improvement in cotton meristem transformation. In vitro cytokinin levels in culture media were tenfold higher in the virGN54D strain, and approximately 30-fold higher in the virGI77V strain, in the absence of acetosyringone induction, compared to the wild type strain. The cytokinin level in the virGN54D strain is further increased upon acetosyringone induction, while the cytokinin level in the virGI77V mutant is decreased by induction, suggesting that different tzs gene expression regulation mechanisms are present in the two virG mutant strains. Based on these data, we suggest that the increased cytokinin levels play a major role in increasing Agrobacterium attachment and stimulating localized division of the attached plant cells.

  20. FAILURE MODE AND CONSTITUTIVE MODEL OF PLAIN HIGH-STRENGTH HIGH-PERFORMANCE CONCRETE UNDER BIAXIAL COMPRESSION AFTER EXPOSURE TO HIGH TEMPERATURES

    Institute of Scientific and Technical Information of China (English)

    Zhenjun He; Yupu Song

    2008-01-01

    An orthotropic constitutive relationship with temperature parameters for plain highstrength high-performance concrete (HSHPC) under biaxial compression is developed. It is based on the experiments performed for characterizing the strength and deformation behavior at two strength levels of HSHPC at 7 different stress ratios including α = σ2 : σ3 = 0.00 : -1, -0.20 : -1, -0.30 : -1, -0.40 : -1, -0.50 : -1, -0.75 : -1, -1.00 : -1, after the exposure to normal and high temperatures of 20, 200, 300, 400, 500 and 600℃, and using a large static-dynamic true triaxial machine. The biaxial tests were performed on 100 mm × 100 mm × 100 mm cubic specimens, and friction-reducing pads were used consisting of three layers of plastic membrane with glycerine in-between for the compressive loading plane. Based on the experimental results, failure modes of HSHPC specimens were described. The principal static compressive strengths, strains at the peak stress and stress-strain curves were measured; and the influence of the temperature and stress ratios on them was also analyzed. The experimental results showed that the uniaxial compressive strength of plain HSHPC after exposure to high temperatures does not decrease dramatically with the increase of temperature. The ratio of the biaxial to its uniaxial compressive strength depends on the stress ratios and brittleness-stiffness of HSHPC after exposure to different temperature levels. Comparison of the stress-strain results obtained from the theoretical model and the experimental data indicates good agreement.

  1. Mitochondria of a human multidrug-resistant hepatocellular carcinoma cell line constitutively express inducible nitric oxide synthase in the inner membrane.

    Science.gov (United States)

    Fantappiè, Ornella; Sassoli, Chiara; Tani, Alessia; Nosi, Daniele; Marchetti, Serena; Formigli, Lucia; Mazzanti, Roberto

    2015-06-01

    Mitochondria play a crucial role in pathways of stress conditions. They can be transported from one cell to another, bringing their features to the cell where they are transported. It has been shown in cancer cells overexpressing multidrug resistance (MDR) that mitochondria express proteins involved in drug resistance such as P-glycoprotein (P-gp), breast cancer resistant protein and multiple resistance protein-1. The MDR phenotype is associated with the constitutive expression of COX-2 and iNOS, whereas celecoxib, a specific inhibitor of COX-2 activity, reverses drug resistance of MDR cells by releasing cytochrome c from mitochondria. It is possible that COX-2 and iNOS are also expressed in mitochondria of cancer cells overexpressing the MDR phenotype. This study involved experiments using the human HCC PLC/PRF/5 cell line with and without MDR phenotype and melanoma A375 cells that do not express the MDR1 phenotype but they do iNOS. Western blot analysis, confocal immunofluorescence and immune electron microscopy showed that iNOS is localized in mitochondria of MDR1-positive cells, whereas COX-2 is not. Low and moderate concentrations of celecoxib modulate the expression of iNOS and P-gp in mitochondria of MDR cancer cells independently from inhibition of COX-2 activity. However, A375 cells that express iNOS also in mitochondria, were not MDR1 positive. In conclusion, iNOS can be localized in mitochondria of HCC cells overexpressing MDR1 phenotype, however this phenomenon appears independent from the MDR1 phenotype occurrence. The presence of iNOS in mitochondria of human HCC cells phenotype probably concurs to a more aggressive behaviour of cancer cells. © 2015 The Authors. Journal of Cellular and Molecular Medicine published by John Wiley & Sons Ltd and Foundation for Cellular and Molecular Medicine.

  2. Listeria-vectored vaccine expressing the Mycobacterium tuberculosis 30 kDa major secretory protein via the constitutively active prfA* regulon boosts BCG efficacy against tuberculosis.

    Science.gov (United States)

    Jia, Qingmei; Dillon, Barbara Jane; Masleša-Galić, Saša; Horwitz, Marcus A

    2017-06-19

    A potent vaccine against tuberculosis, one of the world's deadliest diseases, is needed to enhance the immunity of people worldwide, most of whom have been vaccinated with the partially effective BCG vaccine. Here we investigate novel live attenuated recombinant Listeria monocytogenes (rLm) vaccines expressing the Mycobacterium tuberculosis (Mtb) 30 kDa major secretory protein (r30/Ag85B) (rLm30) as heterologous booster vaccines in animals primed with BCG. Using three attenuated Lm vectors, rLm ΔactA (LmI), rLm ΔactA ΔinlB (LmII), and rLm ΔactA ΔinlBprfA* (LmIII), we constructed five rLm30 vaccine candidates expressing the r30 linked in-frame to the Lm Listeriolycin O signal sequence and driven by the hly promoter (h30) or linked in-frame to the ActA N-terminus and driven by the actA promoter (a30). All five rLm30 vaccines secreted r30 in broth and macrophages; while rLm expressing r30 via a constitutively active prfA* regulon (rLmIII/a30) expressed the greatest amount of r30 in broth culture, all five rLm vaccines expressed equivalent amounts of r30 in infected macrophages. In comparative studies, boosting BCG-immunized mice with rLmIII/a30 induced the strongest antigen-specific T-cell responses, including splenic and lung polyfunctional CD4+ T-cells expressing the three cytokines of interferon-gamma (IFN-γ), tumor necrosis factor-alpha (TNF-α), and interleukin-2 (IL-2) (P 2017 American Society for Microbiology.

  3. The TLR4 D299G and T399I SNPs are constitutively active to up-regulate expression of Trif-dependent genes.

    Directory of Open Access Journals (Sweden)

    Georgina L Hold

    Full Text Available Dysregulated Toll-Like Receptor (TLR signalling and genetic polymorphisms in these proteins are linked to many human diseases. We investigated TLR4 functional variants D299G and T399I to assess the impact on LPS-induced responsiveness in comparison to wild-type TLR4. The mechanism by which this occurs in unclear as these SNPs do not lie within the lipid A binding domain or dimerisation sites of the LPS-TLR4/MD2 receptor complexes. Transfection of TLR4D299G, TLR4T399I or TLR4D299G. T399I into HEK cells resulted in constitutive activation of an NF-κB reporter gene and a blunting of the LPS-induced reporter activation compared to WT-TLR4. Unstimulated human monocyte/macrophages, from patients with the D299G and T399I SNPs demonstrated a downregulation of many genes, particularly Tram/Trif signalling pathway constitutents compared to the TLR4 wild-type subjects supporting the concept of basal receptor activity. Monocyte/macrophages from carriers of the TLR4 D299G and T399I polymorphisms stimulated with LPS showed >6 fold lower levels of NF-κB and ∼12 fold higher IFN-β gene expression levels compared to wild-type subjects (P<0.05; MWU test and dramatically altered resultant cytokine profiles. We conclude that these TLR4 SNPs affect constitutive receptor activity which impacts on the hosts ability to respond to LPS challenge leading to a dysregulated sub-optimal immune response to infection.

  4. Combined high-energy synchrotron X-ray diffraction and computed tomography to characterize constitutive behavior of silica sand

    Energy Technology Data Exchange (ETDEWEB)

    Cil, Mehmet B., E-mail: mcil@utk.edu [Department of Civil and Environmental Engineering, University of Tennessee, Knoxville, TN 37996 (United States); Alshibli, Khalid, E-mail: Alshibli@utk.edu [Department of Civil and Environmental Engineering, University of Tennessee, Knoxville, TN 37996 (United States); Kenesei, Peter, E-mail: kenesei@aps.anl.gov [Argonne National Laboratory, Argonne, IL (United States); Lienert, Ulrich, E-mail: ulrich.lienert@desy.de [Deutsches Elektronen-Synchrotron, DESY Photon Science, Hamburg (Germany)

    2014-04-01

    The deformation behavior of silica sand particles under one-dimensional (1D) loading compression was investigated using nondestructive 3D synchrotron micro-computed tomography (SMT) and three dimensional X-ray diffraction (3DXRD). High-resolution SMT images were used to monitor particle-to-particle interactions, and the onset and propagation of fracture mechanism in a column composed of three silica sand particles. Particle-averaged lattice strain tensors within individual sand particles were measured using the 3DXRD technique and were then used to calculate the stress tensor components via the general elastic stress–strain relationship. The normal stress component in the axial direction (σ{sub zz}) exhibited a nearly linear increasing trend in all sand particles. Shear stress components were in general small relative to the normal stress components and displayed no systematic trend. Knowing lattice strains, stresses, and particle kinematic behavior, one can formulate and develop a micromechanics-based constitutive model to fully characterize strength properties and deformation characteristics of granular materials.

  5. Constitutive expression of RyhB regulates the heme biosynthesis pathway and increases the 5-aminolevulinic acid accumulation in Escherichia coli.

    Science.gov (United States)

    Li, Fangfang; Wang, Yang; Gong, Kai; Wang, Qian; Liang, Quanfeng; Qi, Qingsheng

    2014-01-01

    In the current study, the small RNA ryhB, which regulates the metabolism of iron in Escherichia coli, was constitutively expressed in engineered E. coli DALA. The resulting strain E. coli DALRA produced 16% more 5-aminolevulinic acid (ALA) than the parent strain E. coli DALA in batch fermentation. Meanwhile, we found that addition of iron in the medium increased heme formation and reduced ALA yield, whereas the presence of iron chelator in the medium decreased heme concentration and increased the ALA production efficiency (ALA yield per OD600). The qRT-PCR analysis showed that the mRNA levels of hemB and hemH were also decreased as well as the known RyhB target genes of acnAB, sdhAB, fumA, and cydAB in E. coli DALRA. These results indicated that small RNA can be used as a tool for regulating ALA accumulation in E. coli.

  6. Methadone induces the expression of hepatic drug-metabolizing enzymes through the activation of pregnane X receptor and constitutive androstane receptor.

    Science.gov (United States)

    Tolson, Antonia H; Li, Haishan; Eddington, Natalie D; Wang, Hongbing

    2009-09-01

    Methadone (MD) is the most established substance abuse pharmacotherapy of choice for the management of heroin dependence. To date, drug-drug interactions involving MD have been characterized asymmetrically among existing reports, which describe how other drugs affect the metabolic or pharmacokinetic profiles of MD; however, limited information is available regarding the potential for MD to influence similar fates of coadministered drugs. Moreover, little to no mechanistic evidence has been explored. Here, we show that MD induces hepatic drug-metabolizing enzymes (DMEs) through the activation of pregnane X receptor (PXR) and constitutive androstane receptor (CAR). Real-time polymerase chain reaction analysis of human hepatocyte cultures revealed that MD induces the mRNA expression of CYP2B6, CYP3A4, UGT1A1, and multidrug resistance 1 in a concentration-related manner, with the maximal induction of CYP2B6 challenging that of the induction by rifampicin. Furthermore, MD-mediated induction of CYP2B6 and CYP3A4 proteins was observed in Western blot analysis. In cell-based reporter assays, MD significantly increased human (h) PXR-mediated CYP2B6 reporter activities but exhibited minimal effect on hCAR activation as a result of the constitutive activity of hCAR in HepG2 cells. Further studies revealed that treatment with MD resulted in significant nuclear accumulation of adenovirus/enhanced yellow fluorescent protein tagged-hCAR in human hepatocytes, which has been regarded as the initial step of CAR activation. Additional analysis of the two enantiomers of MD, R-(-)-MD (active) and S-(+)-MD (inactive), indicates the lack of stereoselectivity pertaining to MD-mediated DME induction. Overall, our results show that MD induces the hepatic expression of multiple DMEs by activating PXR- and CAR-mediated pathways.

  7. Improved shoot regeneration, salinity tolerance and reduced fungal susceptibility in transgenic tobacco constitutively expressing PR-10a gene

    Directory of Open Access Journals (Sweden)

    Parinita eAgarwal

    2016-02-01

    Full Text Available Plants in ecosystems are simultaneously exposed to abiotic and biotic stresses, which restrict plant growth and development. The complex responses to these stresses are largely regulated by plant hormones, which in turn, orchestrate the different biochemical and molecular pathways to manoeuvre stress tolerance. The PR-10 protein family is reported to be involved in defence regulation, stress response and plant growth and development. The JcPR-10a overexpression resulted in increased number of shoot buds in tobacco (Nicotiana tabacum, which could be due to high cytokinin to auxin ratio in the transgenics. The docking analysis shows the binding of three BAP molecules at the active sites of JcPR-10a protein. JcPR-10a transgenics showed enhanced salt tolerance, as was evident by increased germination rate, shoot and root length, relative water content, proline, soluble sugar and amino acid content under salinity. Interestingly, the transgenics also showed enhanced endogenous cytokinin level as compared to WT, which, further increased with salinity. Exposure of gradual salinity resulted in increased stomatal conductance, water use efficiency, photosynthesis rate and reduced transpiration rate. Furthermore, the transgenics also showed enhanced resistance against Macrophomina fungus. Thus, JcPR-10a might be working in co-ordination with cytokinin signalling in mitigating the stress induced damage by regulating different stress signalling pathways, leading to enhanced stress tolerance.

  8. Experimental Validation of Two-dimensional Finite Element Method for Simulating Constitutive Response of Polycrystals During High Temperature Plastic Deformation

    Science.gov (United States)

    Agarwal, Sumit; Briant, Clyde L.; Krajewski, Paul E.; Bower, Allan F.; Taleff, Eric M.

    2007-04-01

    A finite element method was recently designed to model the mechanisms that cause superplastic deformation (A.F. Bower and E. Wininger, A Two-Dimensional Finite Element Method for Simulating the Constitutive Response and Microstructure of Polycrystals during High-Temperature Plastic Deformation, J. Mech. Phys. Solids, 2004, 52, p 1289-1317). The computations idealize the solid as a collection of two-dimensional grains, separated by sharp grain boundaries. The grains may deform plastically by thermally activated dislocation motion, which is modeled using a conventional crystal plasticity law. The solid may also deform by sliding on the grain boundaries, or by stress-driven diffusion of atoms along grain boundaries. The governing equations are solved using a finite element method, which includes a front-tracking procedure to monitor the evolution of the grain boundaries and surfaces in the solid. The goal of this article is to validate these computations by systematically comparing numerical predictions to experimental measurements of the elevated-temperature response of aluminum alloy AA5083 (M.-A. Kulas, W.P. Green, E.M. Taleff, P.E. Krajewski, and T.R. McNelley, Deformation Mechanisms in Superplastic AA5083 materials. Metall. Mater. Trans. A, 2005, 36(5), p 1249-1261). The experimental work revealed that a transition occurs from grain-boundary sliding to dislocation (solute-drag) creep at approximately 0.001/s for temperatures between 425 and 500 °C. In addition, increasing the grain size from 7 to 10 μm decreased the transition to significantly lower strain rates. Predictions from the finite element method accurately predict the effect of grain size on the transition in deformation mechanisms.

  9. Stability of High Slope Interbedded Strata with Low Dip Angle Constituted by Soft and Hard Rock Mass

    Institute of Scientific and Technical Information of China (English)

    邓荣贵; 周德培; 张倬元

    2002-01-01

    Slopes consisting of interbedded strata of soft and hard rock mass, such as purplish red mudstone and grey-brown arkosic sandstone of Jurassic age, are very common in Sichuan basin of China. The mudstone is soft while the sandstone is hard and contains many opening or closing joints with a high dip angle. Some are nearly parallel and the others are nearly decussated with the trend of the slopes. Many natural slopes are in deformation or sliding because of those reasons. The stability of cutting slopes and supporting method to be taken for their stability in civil engineering are important. In this paper, the stability and deformation of the slopes are studied. The methods of analysis and support design principle are analyzed also. Finally, the method put forward is applied to study Fengdian high cutting slope in Sichuan section of the express way from Chengdu to Shanghai. The results indicate that the method is effective.

  10. Transgenic salt-tolerant sugar beet (Beta vulgaris L.) constitutively expressing an Arabidopsis thaliana vacuolar Na/H antiporter gene, AtNHX3, accumulates more soluble sugar but less salt in storage roots.

    Science.gov (United States)

    Liu, Hua; Wang, Qiuqing; Yu, Mengmeng; Zhang, Yanyan; Wu, Yingbao; Zhang, Hongxia

    2008-09-01

    In Arabidopsis thaliana, six vacuolar Na(+)/H(+) antiporters (AtNHX1-6) were identified. Among them, AtNHX1, 2 and 5 are functional Na(+)/H(+) antiporters with the most abundant expression levels in seedling shoots and roots. However, the expression of AtNHX3 in Arabidopsis can only be detected by RT-PCR, and its physiological function still remains unclear. In this work, we demonstrate that constitutive expression of AtNHX3 in sugar beet (Beta vulgaris L.) conferred augmented resistance to high salinity on transgenic plants. In the presence of 300 or 500 mm NaCl, transgenic plants showed very high potassium accumulation in the roots and storage roots. Furthermore, the transcripts of sucrose phosphate synthase (SPS), sucrose synthase (SS) and cell wall sucrose invertase (SI) genes were maintained in transgenic plants. The accumulation of soluble sugar in the storage roots of transgenic plants grown under high salt stress condition was also higher. Our results implicate that AtNHX3 is also a functional antiporter responsible for salt tolerance by mediating K(+)/H(+) exchange in higher plants. The salt accumulation in leaves but not in the storage roots, and the increased yield of storage roots with enhanced constituent soluble sugar contents under salt stress condition demonstrate a great potential use of this gene in improving the quality and yield of crop plants.

  11. The DNA puff BhB10-1 gene is differentially expressed in various tissues of Bradysia hygida late larvae and constitutively transcribed in transgenic Drosophila

    Directory of Open Access Journals (Sweden)

    N. Monesi

    2001-07-01

    Full Text Available We extended the characterization of the DNA puff BhB10-1 gene of Bradysia hygida by showing that, although its mRNA is detected only at the end of the fourth larval instar, BhB10-1 expression is not restricted to the salivary gland, the tissue in which this gene is amplified. Different amounts of BhB10-1 mRNA were detected in other larval tissues such as gut, Malpighian tubules, fat body, brain and cuticle, suggesting that this gene is expressed differentially in the various tissues analyzed. Analysis of transgenic Drosophila carrying the BhB10-1 transcription unit and flanking sequences revealed that the tested fragment promotes transcription in a constitutive manner. We suggest that either cis-regulatory elements are missing in the transgene or factors that temporally regulate the BhB10-1 gene in B. hygida are not conserved in Drosophila.

  12. A constitutively expressed pair of rpoE2-chrR2 in Azospirillum brasilense Sp7 is required for survival under antibiotic and oxidative stress.

    Science.gov (United States)

    Gupta, Namrata; Kumar, Santosh; Mishra, Mukti Nath; Tripathi, Anil Kumar

    2013-02-01

    Extracytoplasmic function (ECF) sigma factors (σ(E)) are known to bring about changes in gene expression to enable bacteria to adapt to different stresses. The Azospirillum brasilense Sp245 genome harbours nine genes encoding σ(E), of which two are adjacent to the genes encoding ChrR-type zinc-binding anti-sigma (ZAS) factors. We describe here the role and regulation of a new pair of rpoE-chrR, which was found in the genome of A. brasilense Sp7 in addition to the previously described rpoE-chrR pair (designated rpoE1-chrR1). The rpoE2-chrR2 pair is also cotranscribed, and their products show protein-protein interaction. The -10 and -35 promoter elements of rpoE2-chrR2 and rpoE1-chrR1 were similar but not identical. Unlike the promoter of rpoE1-chrR1, the rpoE2-chrR2 promoter was neither autoregulated nor induced by oxidative stress. Inactivation of chrR2 or overexpression of rpoE2 in A. brasilense Sp7 resulted in an overproduction of carotenoids. It also conferred resistance to oxidative stresses and antibiotics. By controlling the synthesis of carotenoids, initiation and elongation of translation, protein folding and purine biosynthesis, RpoE2 seems to play a crucial role in preventing and repairing the cellular damage caused by oxidative stress. Lack of autoregulation and constitutive expression of rpoE2-chrR2 suggest that RpoE2-ChrR2 may provide a rapid mechanism to cope with oxidative stress, wherein singlet oxygen ((1)O(2))-mediated dissociation of the RpoE2-ChrR2 complex might release RpoE2 to drive the expression of its target genes.

  13. Nucleotide sequence and transcript organization of a region of the vaccinia virus genome which encodes a constitutively expressed gene required for DNA replication.

    Science.gov (United States)

    Roseman, N A; Hruby, D E

    1987-05-01

    A vaccinia virus (VV) gene required for DNA replication has been mapped to the left side of the 16-kilobase (kb) VV HindIII D DNA fragment by marker rescue of a DNA- temperature-sensitive mutant, ts17, using cloned fragments of the viral genome. The region of VV DNA containing the ts17 locus (3.6 kb) was sequenced. This nucleotide sequence contains one complete open reading frame (ORF) and two incomplete ORFs reading from left to right. Analysis of this region at early times revealed that transcription from the incomplete upstream ORF terminates coincidentally with the complete ORF encoding the ts17 gene product, which is directly downstream. The predicted proteins encoded by this region correlate well with polypeptides mapped by in vitro translation of hybrid-selected early mRNA. The nucleotide sequences of a 1.3-kb BglII fragment derived from ts17 and from two ts17 revertants were also determined, and the nature of the ts17 mutation was identified. S1 nuclease protection studies were carried out to determine the 5' and 3' ends of the transcripts and to examine the kinetics of expression of the ts17 gene during viral infection. The ts17 transcript is present at both early and late times postinfection, indicating that this gene is constitutively expressed. Surprisingly, the transcriptional start throughout infection occurs at the proposed late regulatory element TAA, which immediately precedes the putative initiation codon ATG. Although the biological activity of the ts17-encoded polypeptide was not identified, it was noted that in ts17-infected cells, expression of a nonlinked VV immediate-early gene (thymidine kinase) was deregulated at the nonpermissive temperature. This result may indicate that the ts17 gene product is functionally required at an early step of the VV replicative cycle.

  14. Expressing Constitutively Active Rheb in Adult Neurons after a Complete Spinal Cord Injury Enhances Axonal Regeneration beyond a Chondroitinase-Treated Glial Scar.

    Science.gov (United States)

    Wu, Di; Klaw, Michelle C; Connors, Theresa; Kholodilov, Nikolai; Burke, Robert E; Tom, Veronica J

    2015-08-05

    After a spinal cord injury (SCI), CNS axons fail to regenerate, resulting in permanent deficits. This is due to: (1) the presence of inhibitory molecules, e.g., chondroitin sulfate proteoglycans (CSPG), in the glial scar at the lesion; and (2) the diminished growth capacity of adult neurons. We sought to determine whether expressing a constitutively active form of the GTPase Rheb (caRheb) in adult neurons after a complete SCI in rats improves intrinsic growth potential to result in axon regeneration out of a growth-supportive peripheral nerve grafted (PNG) into the SCI cavity. We also hypothesized that treating the glial scar with chondroitinase ABC (ChABC), which digests CSPG, would further allow caRheb-transduced neurons to extend axons across the distal graft interface. We found that targeting this pathway at a clinically relevant post-SCI time point improves both sprouting and regeneration of axons. CaRheb increased the number of axons, but not the number of neurons, that projected into the PNG, indicative of augmented sprouting. We also saw that caRheb enhanced sprouting far rostral to the injury. CaRheb not only increased growth rostral and into the graft, it also resulted in significantly more regrowth of axons across a ChABC-treated scar into caudal spinal cord. CaRheb(+) neurons had higher levels of growth-associated-43, suggestive of a newly identified mechanism for mTOR-mediated enhancement of regeneration. Thus, we demonstrate for the first time that simultaneously addressing intrinsic and scar-associated, extrinsic impediments to regeneration results in significant regrowth beyond an extremely challenging, complete SCI site. After spinal cord injury (SCI), CNS axons fail to regenerate, resulting in permanent deficits. This is due to the diminished growth capacity of adult neurons and the presence of inhibitory molecules in the scar at the lesion. We sought to simultaneously counter both of these obstacles to achieve more robust regeneration after

  15. Even Lesbian Youths or Those Presumed to Be Lesbians Are Protected by the Constitution of Uganda--But to a Limited Extent: Rules the High Court

    Science.gov (United States)

    Mujuzi, Jamil Ddamulira

    2009-01-01

    The Ugandan Penal Code criminalizes same-sex relationships. The author analyzes the Ugandan High Court decision where the judge relied on the Constitution and international human rights instruments to hold that law enforcement officers must respect the rights to privacy and human dignity even of those people presumed to be in same-sex…

  16. Even Lesbian Youths or Those Presumed to Be Lesbians Are Protected by the Constitution of Uganda--But to a Limited Extent: Rules the High Court

    Science.gov (United States)

    Mujuzi, Jamil Ddamulira

    2009-01-01

    The Ugandan Penal Code criminalizes same-sex relationships. The author analyzes the Ugandan High Court decision where the judge relied on the Constitution and international human rights instruments to hold that law enforcement officers must respect the rights to privacy and human dignity even of those people presumed to be in same-sex…

  17. A thermal-mechanical constitutive model for b-HMX single crystal and cohesive interface under dynamic high pressure loading

    Institute of Scientific and Technical Information of China (English)

    2010-01-01

    Due to the significant thermal-mechanical effects during hot spot formation in PBX explosives,a thermodynamic constitutive model has been constructed for HMX anisotropic single crystal subjected to dynamic impact loading. The crystal plasticity model based on dislocation dynamics theory was employed to describe the anisotropic plastic behavior along the preferential slip systems. A modified equation of state (EOS) was introduced into the constitutive equations through the decomposing stress tensor and the nonlinear elasticity for materials was taken into account. The one-dimensional strain impact simulations for HMX single crystal and quasi-bicrystal were performed respectively,in which the cohesive elements were inserted over the interface areas for the latter. The predicted particle velocities for the single crystal sample agreed well with the experimental results in the literature. Furthermore,the effects of crystal orientations,interface,misorientations on localized strain,stress and temperature distributions were predicted and discussed.

  18. Recurrent BCAM-AKT2 fusion gene leads to a constitutively activated AKT2 fusion kinase in high-grade serous ovarian carcinoma

    Science.gov (United States)

    Kannan, Kalpana; Coarfa, Cristian; Chao, Pei-Wen; Luo, Liming; Wang, Yan; Brinegar, Amy E.; Hawkins, Shannon M.; Milosavljevic, Aleksandar; Matzuk, Martin M.; Yen, Laising

    2015-01-01

    High-grade serous ovarian cancer (HGSC) is among the most lethal forms of cancer in women. Excessive genomic rearrangements, which are expected to create fusion oncogenes, are the hallmark of this cancer. Here we report a cancer-specific gene fusion between BCAM, a membrane adhesion molecule, and AKT2, a key kinase in the PI3K signaling pathway. This fusion is present in 7% of the 60 patient cancers tested, a significant frequency considering the highly heterogeneous nature of this malignancy. Further, we provide direct evidence that BCAM-AKT2 is translated into an in-frame fusion protein in the patient’s tumor. The resulting AKT2 fusion kinase is membrane-associated, constitutively phosphorylated, and activated as a functional kinase in cells. Unlike endogenous AKT2, whose activity is tightly regulated by external stimuli, BCAM-AKT2 escapes the regulation from external stimuli. Moreover, a BCAM-AKT2 fusion gene generated via chromosomal translocation using the CRISPR/Cas9 system leads to focus formation in both OVCAR8 and HEK-293T cell lines, suggesting that BCAM-AKT2 is oncogenic. Together, the results indicate that BCAM-AKT2 expression is a new mechanism of AKT2 kinase activation in HGSC. BCAM-AKT2 is the only fusion gene in HGSC that is proven to translate an aberrant yet functional kinase fusion protein with oncogenic properties. This recurrent genomic alteration is a potential therapeutic target and marker of a clinically relevant subtype for tailored therapy of HGSC. PMID:25733895

  19. Constitutive expression of the DUR1,2 gene in an industrial yeast strain to minimize ethyl carbamate production during Chinese rice wine fermentation.

    Science.gov (United States)

    Wu, Dianhui; Li, Xiaomin; Lu, Jian; Chen, Jian; Zhang, Liang; Xie, Guangfa

    2016-01-01

    Urea and ethanol are the main precursors of ethyl carbamate (EC) in Chinese rice wine. During fermentation, urea is generated from arginine by arginase in Saccharomyces cerevisiae, and subsequently cleaved by urea amidolyase or directly transported out of the cell into the fermentation liquor, where it reacts with ethanol to form EC. To reduce the amount of EC in Chinese rice wine, we metabolically engineered two yeast strains, N85(DUR1,2) and N85(DUR1,2)-c, from the wild-type Chinese rice wine yeast strain N85. Both new strains were capable of constitutively expressing DUR1,2 (encodes urea amidolyase) and thus enhancing urea degradation. The use of N85(DUR1,2) and N85(DUR1,2)-c reduced the concentration of EC in Chinese rice wine fermented on a small-scale by 49.1% and 55.3%, respectively, relative to fermentation with the parental strain. All of the engineered strains showed good genetic stability and minimized the production of urea during fermentation, with no exogenous genes introduced during genetic manipulation, and were therefore suitable for commercialization to increase the safety of Chinese rice wine.

  20. Cloning, sequencing and expression analysis of cDNA encoding a constitutive heat shock protein 70 (HSC70) in Fenneropenaeus chinensis

    Institute of Scientific and Technical Information of China (English)

    JIAO Chuanzhen; WANG Zaizhao; LI Fuhua; ZHANG Chengsong; XIANG Jianhai

    2004-01-01

    The cDNA encoding hsc70 of Chinese shrimp Fenneropenaeus chinensis was cloned from hepatopancreas by RT-PCR based on its EST sequence. The full length cDNA of 2090 bp contained an open reading frame of 1956 nucleotides and partial 5′- and 3′-untranslated region(5′- and 3′-UTR). PCR amplification and sequencing analysis showed the existence of introns in the region of 1-547 bp, but they did not exist in the region of 548-2090 bp of hsc70 cDNA. When the deduced 652 amino acid sequence of HSC70 was compared with the members of HSP70 family from other organisms, the results showed 85.9% similarity with HSC71 from Oncorhynchus mykiss and HSC70 from Homo sapiens. It also exhibited 85.8% similarity with HSP70 from Mus musculu and 85.4% with HSC70 from Manduca sexta. Expression analysis showed that hsc70 mRNA was espressed constitutively in hepatopancreas, muscle, eyestalks, haemocytes, heart, ovary, intestine and gills in Fenneropenaeus chinensis. No difference could be detected on hsc70 mRNA level in muscle between heat-shocked and control animals.

  1. A set of fluorescent protein-based markers expressed from constitutive and arbuscular mycorrhiza-inducible promoters to label organelles, membranes and cytoskeletal elements in Medicago truncatula.

    Science.gov (United States)

    Ivanov, Sergey; Harrison, Maria J

    2014-12-01

    Medicago truncatula is widely used for analyses of arbuscular mycorrhizal (AM) symbiosis and nodulation. To complement the genetic and genomic resources that exist for this species, we generated fluorescent protein fusions that label the nucleus, endoplasmic reticulum, Golgi apparatus, trans-Golgi network, plasma membrane, apoplast, late endosome/multivesicular bodies (MVB), transitory late endosome/ tonoplast, tonoplast, plastids, mitochondria, peroxisomes, autophagosomes, plasmodesmata, actin, microtubules, periarbuscular membrane (PAM) and periarbuscular apoplastic space (PAS) and expressed them from the constitutive AtUBQ10 promoter and the AM symbiosis-specific MtBCP1 promoter. All marker constructs showed the expected expression patterns and sub-cellular locations in M. truncatula root cells. As a demonstration of their utility, we used several markers to investigate AM symbiosis where root cells undergo major cellular alterations to accommodate their fungal endosymbiont. We demonstrate that changes in the position and size of the nuclei occur prior to hyphal entry into the cortical cells and do not require DELLA signaling. Changes in the cytoskeleton, tonoplast and plastids also occur in the colonized cells and in contrast to previous studies, we show that stromulated plastids are abundant in cells with developing and mature arbuscules, while lens-shaped plastids occur in cells with degenerating arbuscules. Arbuscule development and secretion of the PAM creates a periarbuscular apoplastic compartment which has been assumed to be continuous with apoplast of the cell. However, fluorescent markers secreted to the periarbuscular apoplast challenge this assumption. This marker resource will facilitate cell biology studies of AM symbiosis, as well as other aspects of legume biology. © 2014 The Authors The Plant Journal © 2014 John Wiley & Sons Ltd.

  2. Constitutive CD40L expression on B cells prematurely terminates germinal center response and leads to augmented plasma cell production in T cell areas.

    Science.gov (United States)

    Bolduc, Anna; Long, Eugene; Stapler, Dale; Cascalho, Marilia; Tsubata, Takeshi; Koni, Pandelakis A; Shimoda, Michiko

    2010-07-01

    CD40/CD40L engagement is essential to T cell-dependent B cell proliferation and differentiation. However, the precise role of CD40 signaling through cognate T-B interaction in the generation of germinal center and memory B cells is still incompletely understood. To address this issue, a B cell-specific CD40L transgene (CD40LBTg) was introduced into mice with B cell-restricted MHC class II deficiency. Using this mouse model, we show that constitutive CD40L expression on B cells alone could not induce germinal center differentiation of MHC class II-deficient B cells after immunization with T cell-dependent Ag. Thus, some other MHC class II-dependent T cell-derived signals are essential for the generation of germinal center B cells in response to T cell-dependent Ag. In fact, CD40LBTg mice generated a complex Ag-specific IgG1 response, which was greatly enhanced in early, but reduced in late, primary response compared with control mice. We also found that the frequency of Ag-specific germinal center B cells in CD40LBTg mice was abruptly reduced 1 wk after immunization. As a result, the numbers of Ag-specific IgG1 long-lived plasma cells and memory B cells were reduced. By histology, large numbers of Ag-specific plasma cells were found in T cell areas adjacent to Ag-specific germinal centers of CD40LBTg mice, temporarily during the second week of primary response. These results indicate that CD40L expression on B cells prematurely terminated their ongoing germinal center response and produced plasma cells. Our results support the notion that CD40 signaling is an active termination signal for germinal center reaction.

  3. Isolation and gene expression analysis of Arabidopsis thaliana mutants with constitutive expression of ATL2, an early elicitor-response RING-H2 zinc-finger gene.

    OpenAIRE

    2004-01-01

    Genes with unstable transcripts often encode proteins that play important regulatory roles. ATL2 is a member of a multigene family coding highly related RING-H2 zinc-finger proteins that may function as E3 ubiquitin ligases. ATL2 mRNA accumulation occurs rapidly and transiently after incubation with elicitors of pathogen response. We screened 50,000 M(2) families from a line that carries a fusion of pATL2 to the GUS reporter gene and isolated five mutants, which we named eca (expresión consti...

  4. Loss of aquaporin 3 protein expression constitutes an independent prognostic factor for progression-free survival: an immunohistochemical study on stage pT1 urothelial bladder cancer

    Directory of Open Access Journals (Sweden)

    Otto Wolfgang

    2012-10-01

    Full Text Available Abstract Background Treatment of patients with stage pT1 urothelial bladder cancer (UBC continues to be a challenge due to its unpredictable clinical course. Reliable molecular markers that help to determine appropriate individual treatment are still lacking. Loss of aquaporin (AQP 3 protein expression has previously been shown in muscle-invasive UBC. The aim of the present study was to investigate the prognostic value of AQP3 protein expression with regard to the prognosis of stage pT1 UBC. Method AQP 3 protein expression was investigated by immunohistochemistry in specimens of 87 stage T1 UBC patients, who were diagnosed by transurethral resection of the bladder (TURB and subsequent second resection at a high-volume urological centre between 2002 and 2009. Patients underwent adjuvant instillation therapy with Bacillus Calmette-Guérin (BCG. Loss of AQP3 protein expression was defined as complete absence of the protein within the whole tumour. Expression status was correlated retrospectively with clinicopathological and follow-up data (median: 31 months. Multivariate Cox regression analysis was used to assess the value of AQP3 tumour expression with regard to recurrence-free (RFS, progression-free (PFS and cancer-specific survival (CSS. RFS, PFS and CSS were calculated by Kaplan-Meier analysis and Log rank test. Results 59% of patients were shown to exhibit AQP3-positive tumours, whereas 41% of tumours did not express the marker. Loss of AQP3 protein expression was associated with a statistically significantly worse PFS (20% vs. 72%, p=0.020. This finding was confirmed by multivariate Cox regression analysis (HR 7.58, CI 1.29 – 44.68; p=0.025. Conclusions Loss of AQP3 protein expression in pT1 UBC appears to play a key role in disease progression and is associated with worse PFS. Considering its potential prognostic value, assessment of AQP3 protein expression could be used to help stratify the behavior of patients with pT1 UBC.

  5. Bt rice harbouring cry genes controlled by a constitutive or wound-inducible promoter: protection and transgene expression under Mediterranean field conditions.

    Science.gov (United States)

    Breitler, Jean Christophe; Vassal, Jean Michel; del Mar Catala, Maria; Meynard, Donaldo; Marfà, Victoria; Melé, Enric; Royer, Monique; Murillo, Isabel; San Segundo, Blanca; Guiderdoni, Emmanuel; Messeguer, Joaquima

    2004-09-01

    Seven homozygous transgenic lines of two European commercial cultivars of rice (Ariete (A) and Senia (S)), harbouring the cry1B or cry1Aa Bacillus thuringiensis (Bt) delta-endotoxin genes, were field evaluated for protection from striped stem borer (SSB) (Chilo suppressalis) damage during the 2001 and 2002 summer crop seasons in the Delta de l'Ebre region, Spain. The plant codon-optimized toxin gene was placed under the control of the promoter of either the constitutive ubi1 gene or the wound-inducible mpi gene from maize. Stable, high-level, insecticidal protein accumulation was observed throughout root, leaf and seed tissues of field-grown plants harbouring the cry1B (lines A64.1, A33.1, A3.4 and S98.9) or cry1Aa (lines S05.1 and A19.14) genes under the control of the ubi1 promoter. Conversely, no toxin was detected in unwounded vegetative tissues of the A9.1 line harbouring the cry1B gene controlled by the mpi promoter, indicating that natural environmental stresses did not trigger the activity of the wound-inducible promoter. However, the toxin accumulated at 0.2% total soluble proteins in A9.1 sheath tissue exhibiting brown lesions resulting from SSB damage. The agronomical traits and performance of the transgenic lines were generally comparable with parental controls, except in the two lines accumulating Cry1Aa, which exhibited a high frequency of plants non-true to type. Natural infestation was assisted with manual infestations of L2/L3 SSB larvae in border control plants surrounding the experimental plots, which served as a reservoir for the second-cycle SSB population. The observation of damage (brown lesions and dead hearts) during the crop season and dissection of plants at harvest stage revealed a range of protection amongst the transgenic lines, which was highly consistent with the level of toxin accumulation and with previous experience in greenhouse assays. Lines A3.4 and S05.1 were found to exhibit stable and full protection against SSB attacks

  6. Phase Constitution in Mixed Gd2O3 and B4C by Sintering at High Temperature

    Institute of Scientific and Technical Information of China (English)

    Wu Wenyuan; Xu Jingyu; Bian Xue; Hu Guangyong; Sun Shuchen; Tu Ganfeng

    2005-01-01

    The phase constitution in mixed Gd2O3 and B4C by sintering in graphite tube furnace at the temperature of 100~1489 ℃ in argon atmosphere was studied by means of XRD and TG-DTA. The results show that the impurity C reacts with O in the B4C at the temperature of 367~458 ℃. When the temperature is 800 ℃, Gd2O3 reacts with B4C, and the reaction products include GdB6, GdB4, GdBO3, GdBC and B. GdBC changes into GdB4 at 1200 ℃. When the temperature is 1470 ℃, GdB6 is obtained by the reaction of GdB4 with elemental B. The content of GdB6 is increasing with the prolonging time.

  7. Constitutive Activation of the G-Protein Subunit G[alpha]s within Forebrain Neurons Causes PKA-Dependent Alterations in Fear Conditioning and Cortical "Arc" mRNA Expression

    Science.gov (United States)

    Kelly, Michele P.; Cheung, York-Fong; Favilla, Christopher; Siegel, Steven J.; Kanes, Stephen J.; Houslay, Miles D.; Abel, Ted

    2008-01-01

    Memory formation requires cAMP signaling; thus, this cascade has been of great interest in the search for cognitive enhancers. Given that medications are administered long-term, we determined the effects of chronically increasing cAMP synthesis in the brain by expressing a constitutively active isoform of the G-protein subunit G[alpha]s…

  8. The Na+/H+ exchanger NHE1, but not the Na+, HCO3- cotransporter NBCn1, regulates motility of MCF7 breast cancer cells expressing constitutively active ErbB2

    DEFF Research Database (Denmark)

    Lauritzen, Gitte; Stock, Christian-Martin; Lemaire, Justine;

    2012-01-01

    We and others have shown central roles of the Na(+)/H(+) exchanger NHE1 in cell motility. The aim of this study was to determine the roles of NHE1 and of the Na(+), HCO(3)(-) cotransporter NBCn1 in motility of serum-starved MCF-7 breast cancer cells expressing constitutively active ErbB2 (¿NErbB2...

  9. Cloning, high-level expression, purification and characterization of a ...

    African Journals Online (AJOL)

    Cloning, high-level expression, purification and characterization of a staphylokinase variant, SakøC, ... African Journal of Biotechnology ... Hence in this study, we reported the cloning, high-level expression, purification and characterization of ...

  10. Living with high putrescine: expression of ornithine and arginine biosynthetic pathway genes in high and low putrescine producing poplar cells.

    Science.gov (United States)

    Page, Andrew F; Minocha, Rakesh; Minocha, Subhash C

    2012-01-01

    Arginine (Arg) and ornithine (Orn), both derived from glutamate (Glu), are the primary substrates for polyamine (PA) biosynthesis, and also play important roles as substrates and intermediates of overall N metabolism in plants. Their cellular homeostasis is subject to multiple levels of regulation. Using reverse transcription quantitative PCR (RT-qPCR), we studied changes in the expression of all genes of the Orn/Arg biosynthetic pathway in response to up-regulation [via transgenic expression of mouse Orn decarboxylase (mODC)] of PA biosynthesis in poplar (Populus nigra × maximowiczii) cells grown in culture. Cloning and sequencing of poplar genes involved in the Orn/Arg biosynthetic pathway showed that they have high homology with similar genes in other plants. The expression of the genes of Orn, Arg and PA biosynthetic pathway fell into two hierarchical clusters; expression of one did not change in response to high putrescine, while members of the other cluster showed a shift in expression pattern during the 7-day culture cycle. Gene expression of branch point enzymes (N-acetyl-Glu synthase, Orn aminotransferase, Arg decarboxylase, and spermidine synthase) in the sub-pathways, constituted a separate cluster from those involved in intermediary reactions of the pathway (N-acetyl-Glu kinase, N-acetyl-Glu-5-P reductase, N-acetyl-Orn aminotransferase, N (2)-acetylOrn:N-acetyl-Glu acetyltransferase, N (2)-acetyl-Orn deacetylase, Orn transcarbamylase, argininosuccinate synthase, carbamoylphosphate synthetase, argininosuccinate lyase, S-adenosylmethionine decarboxylase, spermine synthase). We postulate that expression of all genes of the Glu-Orn-Arg pathway is constitutively coordinated and is not influenced by the increase in flux rate through this pathway in response to increased utilization of Orn by mODC; thus the pathway involves mostly biochemical regulation rather than changes in gene expression. We further suggest that Orn itself plays a major role in the

  11. Elasto-Plastic-Creep Constitutive Equation of an Al-Si-Cu High-Pressure Die Casting Alloy for Thermal Stress Analysis

    Science.gov (United States)

    Motoyama, Yuichi; Shiga, Hidetoshi; Sato, Takeshi; Kambe, Hiroshi; Yoshida, Makoto

    2016-11-01

    Accurate simulation of residual stress and deformation is necessary to optimize the design and lifetime of casting components. Therefore, the recovery and strain-rate dependence of the stress-strain curve have been incorporated into empirical constitutive equations to improve the thermal stress analysis accuracy. Nevertheless, these equations present several difficulties related to the determination of material constants and their physical bases. This study suggested an empirical elasto-plastic-creep constitutive equation incorporating these phenomena. To determine the material parameters used in this constitutive equation, this study investigated tensile test methods to obtain stress-strain curves that most closely resemble those during or immediately after casting for the Al-Si-Cu high-pressure die-casting alloy JIS ADC 12 (A383.0), which exhibits natural aging. Results show that solution heat treatment with subsequent cooling to the test temperature should be applied to obtain stress-strain curves used for the thermal stress analysis of high-pressure die casting process of this alloy. The yield stresses obtained using the conventional heating method were 50-64 pct higher than those of the method described above. Therefore, the conventional method is expected to overestimate the overestimation of the predicted residual stress in die castings. Evaluation of the developed equation revealed that it can represent alloy recovery and strain-rate dependence.

  12. Constitutive Cyclic Deformation Behavior in Single-crystal and Directionally Solidified SSME High-pressure Fuel Turbopump Airfoil Materials

    Science.gov (United States)

    Milligan, W. W.; Huron, E. S.; Antolovich, S. D.

    1985-01-01

    The major goal of the project is to correlate mechanical properties with microstructural deformation behavior and to develop models for constitutive response under a variety of monotonic and cyclic loading cycles, temperatures, strain levels, strain rates, and environments. Two alloys are being studied as candidate SSME turbine blade materials. The first is PWA 1480, which is a single-crystal alloy whose nominal composition is reported. The second alloy being studied is D.S. Mar-M 246 + Hf. This is a directionally solidified material, and its nominal composition is also reported. The major areas of interest for the two materials will be slightly different. The single-crystal alloy lends itself well to fundamental deformation studies, since resolved shear stresses on slip planes are all known and only one grain is present. The D.S. material presents an excellent opportunity to study the effects of slightly misaligned grains on deformation behavior. The two materials will be studied by using approximately the same test matrix, so a good degree of direct comparison will also be possible.

  13. Constitutive expression of a barley Fe phytosiderophore transporter increases alkaline soil tolerance and results in iron partitioning between vegetative and storage tissues under stress

    OpenAIRE

    Gómez-Galera, Sonia; Sudhakar, Duraialagaraja; Ana M. Pelacho; Capell, Teresa; Christou, Paul

    2012-01-01

    Cereals have evolved chelation systems to mobilize insoluble iron in the soil, but in rice this process is rather inefficient, making the crop highly susceptible to alkaline soils. We therefore engineered rice to express the barley iron-phytosiderophore transporter (HvYS1), which enables barley plants to take up iron from alkaline soils. A representative transgenic rice line was grown in standard (pH 5.5) or alkaline soil (pH 8.5) to evaluate alkaline tolerance and iron mobilizati...

  14. utilizing constitutional values in constitutional comparison

    African Journals Online (AJOL)

    Administrator

    review, the separation of powers, democratic processes, etc. ... 2 The approach to constitutional comparison followed here, is more extensively dealt with ..... race, national or ethnic origin, color, religion, sex, age or mental or physical disability.

  15. Constitutive oligomerization of human D2 dopamine receptors expressed in Spodoptera frugiperda 9 (Sf9) and in HEK293 cells. Analysis using co-immunoprecipitation and time-resolved fluorescence resonance energy transfer.

    Science.gov (United States)

    Gazi, Lucien; López-Giménez, Juan F; Rüdiger, Martin P; Strange, Philip G

    2003-10-01

    Human D2Long (D2L) and D2Short (D2S) dopamine receptor isoforms were modified at their N-terminus by the addition of a human immunodeficiency virus (HIV) or a FLAG epitope tag. The receptors were then expressed in Spodoptera frugiperda 9 (Sf9) cells using the baculovirus system, and their oligomerization was investigated by means of co-immunoprecipitation and time-resolved fluorescence resonance energy transfer (FRET). [3H]Spiperone labelled D2 receptors in membranes prepared from Sf9 cells expressing epitope-tagged D2L or D2S receptors, with a pKd value of approximately 10. Co-immunoprecipitation using antibodies specific for the tags showed constitutive homo-oligomerization of D2L and D2S receptors in Sf9 cells. When the FLAG-tagged D2S and HIV-tagged D2L receptors were co-expressed, co-immunoprecipitation showed that the two isoforms can also form hetero-oligomers in Sf9 cells. Time-resolved FRET with europium and XL665-labelled antibodies was applied to whole Sf9 cells and to membranes from Sf9 cells expressing epitope-tagged D2 receptors. In both cases, constitutive homo-oligomers were revealed for D2L and D2S isoforms. Time-resolved FRET also revealed constitutive homo-oligomers in HEK293 cells expressing FLAG-tagged D2S receptors. The D2 receptor ligands dopamine, R-(-)propylnorapomorphine, and raclopride did not affect oligomerization of D2L and D2S in Sf9 and HEK293 cells. Human D2 dopamine receptors can therefore form constitutive oligomers in Sf9 cells and in HEK293 cells that can be detected by different approaches, and D2 oligomerization in these cells is not regulated by ligands.

  16. Effect of inducers and culturing processes on laccase synthesis in Phanerochaete chrysosporium NCIM 1197 and the constitutive expression of laccase isozymes

    DEFF Research Database (Denmark)

    Manavalan, Arulmani

    2006-01-01

    Phanerochaete chrysosporium NCIM 1197 constitutively secretes considerable level of extracellular enzyme laccase in defined growth medium. Effect of several inducers on laccase production was attempted and found that copper sulphate alone at 30 mM concentration accelerate the laccase production...

  17. Transgenic maize plants expressing the Totivirus antifungal protein, KP4, are highly resistant to corn smut.

    Science.gov (United States)

    Allen, Aron; Islamovic, Emir; Kaur, Jagdeep; Gold, Scott; Shah, Dilip; Smith, Thomas J

    2011-10-01

    The corn smut fungus, Ustilago maydis, is a global pathogen responsible for extensive agricultural losses. Control of corn smut using traditional breeding has met with limited success because natural resistance to U. maydis is organ specific and involves numerous maize genes. Here, we present a transgenic approach by constitutively expressing the Totivirus antifungal protein KP4, in maize. Transgenic maize plants expressed high levels of KP4 with no apparent negative impact on plant development and displayed robust resistance to U. maydis challenges to both the stem and ear tissues in the greenhouse. More broadly, these results demonstrate that a high level of organ independent fungal resistance can be afforded by transgenic expression of this family of antifungal proteins.

  18. Constitutional Verbosity and Social Trust

    DEFF Research Database (Denmark)

    Bjørnskov, Christian; Voigt, Stefan

    2014-01-01

    A common argument in the trust literature is that high-trust cultures allow efficient commercial contracts to be shorter, covering fewer contingencies. We take this idea to the topic of social contracts. Specifically, we ask whether social trust affects the length and detail of constitutions. Cross......-country estimates suggest that national trust levels are indeed robustly and negatively associated with the length of countries’ constitutions....

  19. Constitutive expression of a barley Fe phytosiderophore transporter increases alkaline soil tolerance and results in iron partitioning between vegetative and storage tissues under stress.

    Science.gov (United States)

    Gómez-Galera, Sonia; Sudhakar, Duraialagaraja; Pelacho, Ana M; Capell, Teresa; Christou, Paul

    2012-04-01

    Cereals have evolved chelation systems to mobilize insoluble iron in the soil, but in rice this process is rather inefficient, making the crop highly susceptible to alkaline soils. We therefore engineered rice to express the barley iron-phytosiderophore transporter (HvYS1), which enables barley plants to take up iron from alkaline soils. A representative transgenic rice line was grown in standard (pH 5.5) or alkaline soil (pH 8.5) to evaluate alkaline tolerance and iron mobilization. Transgenic plants developed secondary tillers and set seeds when grown in standard soil although iron concentration remained similar in leaves and seeds compared to wild type. However, when grown in alkaline soil transgenic plants exhibited enhanced growth, yield and iron concentration in leaves compared to the wild type plants which were severely stunted. Transgenic plants took up iron more efficiently from alkaline soil compared to wild type, indicating an enhanced capacity to increase iron mobility ex situ. Interestingly, all the additional iron accumulated in vegetative tissues, i.e. there was no difference in iron concentration in the seeds of wild type and transgenic plants. Our data suggest that iron uptake from the rhizosphere can be enhanced through expression of HvYS1 and confirm the operation of a partitioning mechanism that diverts iron to leaves rather than seeds, under stress.

  20. Constitutive over-expression of rice ClpD1 protein enhances tolerance to salt and desiccation stresses in transgenic Arabidopsis plants.

    Science.gov (United States)

    Mishra, Ratnesh Chandra; Richa; Grover, Anil

    2016-09-01

    Caseinolytic proteases (Clps) perform the important role of removing protein aggregates from cells, which can otherwise prove to be highly toxic. ClpD system is a two-component protease complex composed of a regulatory ATPase module ClpD and a proteolytic component ClpP. Under desiccation stress condition, rice ClpD1 (OsClpD1) gene encoding for the regulatory subunit, was represented by four variant transcripts differing mainly in the expanse of their N-terminal amino acids. These transcripts were expressed in a differential manner in response to salt, mannitol and polyethylene glycol stresses in rice. Purified OsClpD1.3 protein exhibited intrinsic chaperone activity, shown using citrate synthase as substrate. Arabidopsis (Col-0) plants over-expressing OsClpD1.3 open reading frame downstream to CaMV35S promoter (ClpD1.3 plants) showed higher tolerance to salt and desiccation stresses as compared to wild type plants. ClpD1.3 seedlings also showed enhanced growth during the early stages of seed germination under unstressed, control conditions. The free proline levels and starch breakdown activities were higher in the ClpD1.3 seedlings as compared to the wild type Arabidopsis seedlings. It thus emerges that increasing the potential of ClpD1 chaperoning activity may be of advantage in protection against abiotic stresses. Copyright © 2016 Elsevier Ireland Ltd. All rights reserved.

  1. "Ballet It's Too Whitey": Discursive Hierarchies of High School Dance Spaces and the Constitution of Embodied Feminine Subjectivities

    Science.gov (United States)

    Atencio, Matthew; Wright, Jan

    2009-01-01

    This article investigates (i) how the structuring practices and meanings associated with dance classes at an inner-city American high school operated as institutional spaces (re)producing "dividing practices" that supported racial and classed hierarchies; (ii) how these racist structures were created and maintained relative to dominant notions of…

  2. Constitutive modeling and finite element procedure development for stress analysis of prismatic high temperature gas cooled reactor graphite core components

    Energy Technology Data Exchange (ETDEWEB)

    Mohanty, Subhasish, E-mail: smohanty@anl.gov [Argonne National Laboratory, South Cass Avenue, Argonne, IL 60439 (United States); Majumdar, Saurindranath [Argonne National Laboratory, South Cass Avenue, Argonne, IL 60439 (United States); Srinivasan, Makuteswara [U.S. Nuclear Regulatory Commission, Washington, DC 20555 (United States)

    2013-07-15

    Highlights: • Finite element procedure developed for stress analysis of HTGR graphite component. • Realistic fluence profile and reflector brick shape considered for the simulation. • Also realistic H-451 grade material properties considered for simulation. • Typical outer reflector of a GT-MHR type reactor considered for numerical study. • Based on the simulation results replacement of graphite bricks can be scheduled. -- Abstract: High temperature gas cooled reactors, such as prismatic and pebble bed reactors, are increasingly becoming popular because of their inherent safety, high temperature process heat output, and high efficiency in nuclear power generation. In prismatic reactors, hexagonal graphite bricks are used as reflectors and fuel bricks. In the reactor environment, graphite bricks experience high temperature and neutron dose. This leads to dimensional changes (swelling and or shrinkage) of these bricks. Irradiation dimensional changes may affect the structural integrity of the individual bricks as well as of the overall core. The present paper presents a generic procedure for stress analysis of prismatic core graphite components using graphite reflector as an example. The procedure is demonstrated through commercially available ABAQUS finite element software using the option of user material subroutine (UMAT). This paper considers General Atomics Gas Turbine-Modular Helium Reactor (GT-MHR) as a bench mark design to perform the time integrated stress analysis of a typical reflector brick considering realistic geometry, flux distribution and realistic irradiation material properties of transversely isotropic H-451 grade graphite.

  3. Mouse newborn cells allow highly productive mouse cytomegalovirus replication, constituting a novel convenient primary cell culture system.

    Science.gov (United States)

    Le-Trilling, Vu Thuy Khanh; Trilling, Mirko

    2017-01-01

    Mammalian cell culture is indispensable for most aspects of current biomedical research. Immortalized cell lines are very convenient, but transforming principles (e.g. oncogenic viruses or their oncogenes) can heavily influence the experimental outcome. Primary cells do not share this apparent disadvantage but are more laborious to generate. Certain viruses (e.g. mouse cytomegalovirus) do not replicate efficiently in most transformed cell lines. In the past, such viruses have been routinely propagated on primary mouse embryonic fibroblasts (MEF) established around day 17 (d17) of gestation. According to new regulations of the European Union, experiments using gravid mammals and/or their embryos in the last trimester (>d14 in the case of mice) of gestation do require explicit permission of the local authorities responsible for animal care and use. Applying for such permission is time-consuming and often inflexible. Embryonic fibroblasts could also be produced at earlier time points of pregnancy from younger and smaller embryos. Obviously, this approach consumes more pregnant mice and embryos. Newborn mice are larger thus yielding more cells per sacrificed animal and the new Directive (2010/63/EU) excludes the killing of animals solely for the use of their organs or tissues. We established a convenient protocol to generate adherent mouse newborn cells (MNC). A direct comparison of MNC with MEF revealed that MNC fully recapitulate all tested aspects of a broad panel of virological parameters (plaque size, final titers, viral replication kinetics, viral gene expression, drug and interferon susceptibility as well as species specificity). The herein described approach allows researchers the legal use of primary cells and contributes to the 3R (replace, reduce, refine) guiding principles-especially the 'reduce' aspect-for the use of animals in scientific research. Additionally, it offers the option to directly compare in vitro and in vivo experiments when MNC are

  4. Inhibition of constitutively activated Stat3 correlates with altered Bcl-2/Bax expression and induction of apoptosis in mycosis fungoides tumor cells

    DEFF Research Database (Denmark)

    Nielsen, M; Kaestel, C G; Eriksen, K W

    1999-01-01

    promotor. The decreased ability of Stat3 to bind DNA precedes dynamic alterations in the expression of anti-apoptotic Bcl-2 and pro-apoptotic Bax proteins (decreased Bcl-2 expression and increased Bax expression) and induction of apoptosis. Thus, our data suggest that the involvement of Stat3 in oncogenic...

  5. Ovarian cancer patients at high risk of BRCA mutation: the constitutional genetic characterization does not change prognosis.

    Science.gov (United States)

    Sabatier, Renaud; Lavit, Elise; Moretta, Jessica; Lambaudie, Eric; Noguchi, Tetsuro; Eisinger, François; Cherau, Elisabeth; Provansal, Magali; Livon, Doriane; Rabayrol, Laetitia; Popovici, Cornel; Charaffe-Jauffret, Emmanuelle; Sobol, Hagay; Viens, Patrice

    2016-10-01

    Ovarian neoplasms secondary to germline BRCA mutations had been described to have a more favourable survival. There is only few data concerning the prognosis of non mutated patients presenting clinical features evocative of BRCA alterations. We retrospectively collected data from patients treated in our institution for an invasive ovarian carcinoma between 1995 and 2011. Patients considered at high risk of BRCA mutation were tested for BRCA1/2 germline mutations. We described clinical, pathological and therapeutic features and compared prognosis of BRCA mutation carriers and non-mutated patients. Out of 617 ovarian cancer patients, we identified 104 patients who were considered at high risk of mutation. The 33 mutated patients were more likely to present a personal (33 vs. 10 %, p = 0.003) or a family (42 vs. 24 %, p = 0.06) history of breast/ovarian cancers. BRCA1/2 mutation carriers and wild type patients displayed similar prognosis: median progression-free survival (PFS) of 20.9 versus 37.7 months (p = 0.21); median overall survival (OS) of 151.2 versus 122.5 months (p = 0.52). Personal history of breast cancer increased both PFS [HR = 0.45 (95CI 0.25-0.81)] and OS [HR = 0.35 (95CI 0.16-0.75)]. In multivariate analysis, this parameter was an independent prognostic feature, whereas the identification of a BRCA1/2 mutation was not. In our cohort, all patients at high risk of BRCA mutation share a similar prognosis, whatever is their germline mutation status. Prognosis seems to be more influenced by clinical history than by germline mutations identification. If it is confirmed in larger and independent series, this result suggests that the hypothesis of other BRCA pathway alterations (BRCAness phenotype) deserves to be deeply explored.

  6. Differences in Gene Regulation by Dual Ligands of Nuclear Receptors Constitutive Androstane Receptor (CAR) and Pregnane X Receptor (PXR) in HepG2 Cells Stably Expressing CAR/PXR.

    Science.gov (United States)

    Kanno, Yuichiro; Tanuma, Nobuaki; Yazawa, Saki; Zhao, Shuai; Inaba, Miki; Nakamura, Satoshi; Nemoto, Kiyomitsu; Inouye, Yoshio

    2016-08-01

    The constitutive androstane receptor (CAR) and pregnane X receptor (PXR) regulate various genes involved in xenobiotics and drug metabolism. In many cases, CAR/PXR share ligands termed dual ligands of CAR/PXR. It is difficult to investigate the effect of CAR/PXR dual ligands in cell lines because CAR and PXR expression is scarcely detected in cultured cell lines. Here, we established a tetracycline-inducible human CAR and stably human PXR-overexpressing HepG2 cell line (HepTR/hCAR/hPXR) to examine CAR/PXR dual ligands. In the present study, we investigated the regulation of CYP2B6, CYP2C9, CYP3A4, and UDP-glucuronosyl transferase, which are target genes of CAR/PXR, by dual ligands of CAR/PXR in two transfectants. Activation of CAR and PXR in cells treated with a high dose of CITCO [6-(4-chlorophenyl)-imidazo(2,1-b)thiazole-5-carbaldehyde] or cotreated with rifampicin and tetracycline resulted in synergistic enhancement of CYP3A4, but not CYP2B6, CYP2C9, or UGT1A1, mRNA expression in HepTR/hCAR/hPXR cells. In contrast, this synergistic effect was not observed in HepTR/hCAR cells. These observations were also demonstrated in human primary hepatocytes. Taken together, our results suggest that dual ligands of CAR/PXR show distinct gene regulation patterns by cross-talk between CAR and PXR. Furthermore, the two newly established cell lines are useful tools to investigate dual ligands of CAR/PXR.

  7. A Global Labour Constitution?

    OpenAIRE

    Dukes, R.

    2014-01-01

    This article explores the argument that the idea of the labour constitution, as developed by Hugo Sinzheimer, offers a useful perspective for thinking about labour law today. With reference to the work of Wolfgang Streeck and Karl Polanyi, it highlights the potential benefits of the labour constitution as a framework for analysis. With a view to developing and updating Sinzheimer’s blueprint for a – national – labour constitution, it then engages with two lines of theoretical enquiry into the...

  8. Role of AmpR in the High Expression of the Plasmid-Encoded AmpC β-Lactamase CFE-1.

    Science.gov (United States)

    Nakano, Ryuichi; Nakano, Akiyo; Yano, Hisakazu; Okamoto, Ryoichi

    2017-01-01

    CFE-1 is a unique plasmid-encoded AmpC β-lactamase with the regulator gene ampR. It imparts high resistance to most cephalosporins with constitutive high-level β-lactamase activity. Here, the β-lactamase activities and expression levels of ampC with or without ampR were investigated. Results suggested that the resistance of CFE-1 to cephalosporins is caused by a substitution in AmpR, in which the Asp at position 135 is modified to Ala to allow the constitutive high-level expression (derepression) of ampC.

  9. Low Constitutive Cell Surface Expression of HLA-B Is Caused by a Posttranslational Mechanism Involving Glu180 and Arg239

    DEFF Research Database (Denmark)

    Dellgren, Christoffer; Ekwelum, Vanessa A. C.; Ormhøj, Maria

    2016-01-01

    HLA class I cell surface expression is crucial for normal immune responses, and variability in HLA expression may influence the course of infections. We have previously shown that classical HLA class I expression on many human cell types is biased with greatly reduced expression of HLA-B compared...... with HLA-A in the absence of inflammatory signals. In the search for the mechanisms responsible for this discrepancy, we have recently reported that the regulation is mainly posttranslational and that the C-terminal part of the α2 domain and the α3 domain contain the molecular determinants that explain...

  10. Constitutive over-expression of rice chymotrypsin protease inhibitor gene OCPI2 results in enhanced growth, salinity and osmotic stress tolerance of the transgenic Arabidopsis plants.

    Science.gov (United States)

    Tiwari, Lalit Dev; Mittal, Dheeraj; Chandra Mishra, Ratnesh; Grover, Anil

    2015-07-01

    Protease inhibitors are involved primarily in defense against pathogens. In recent years, these proteins have also been widely implicated in response of plants to diverse abiotic stresses. Rice chymotrypsin protease inhibitor gene OCPI2 is highly induced under salt and osmotic stresses. The construct containing the complete coding sequence of OCPI2 cloned downstream to CaMV35S promoter was transformed in Arabidopsis and single copy, homozygous transgenic lines were produced. The transgenic plants exhibited significantly enhanced tolerance to NaCl, PEG and mannitol stress as compared to wild type plants. Importantly, the vegetative and reproductive growth of transgenic plants under unstressed, control conditions was also enhanced: transgenic plants were more vigorous than wild type, resulting into higher yield in terms of silique number. The RWC values and membrane stability index of transgenic in comparison to wild type plants was higher. Higher proline content was observed in the AtOCPI2 lines, which was associated with higher transcript expression of pyrroline-5-carboxylate synthase and lowered levels of proline dehydrogenase genes. The chymotrypsin protease activities were lower in the transgenic as against wild type plants, under both unstressed, control as well as stressed conditions. It thus appears that rice chymotrypsin protease inhibitor gene OCPI2 is a useful candidate gene for genetic improvement of plants against salt and osmotic stress.

  11. The Constitution by Cell

    Science.gov (United States)

    Greenhut, Stephanie; Jones, Megan

    2010-01-01

    On their visit to the National Archives Experience in Washington, D.C., students in Jenni Ashley and Gay Brock's U.S. history classes at the Potomac School in McLean, Virginia, participated in a pilot program called "The Constitution by Cell." Armed with their cell phones, a basic understanding of the Constitution, and a willingness to participate…

  12. Interpreting the Constitution.

    Science.gov (United States)

    Brennan, William J., Jr.

    1987-01-01

    Discusses constitutional interpretations relating to capital punishment and protection of human dignity. Points out the document's effectiveness in creating a new society by adapting its principles to current problems and needs. Considers two views of the Constitution that lead to controversy over the legitimacy of judicial decisions. (PS)

  13. The Constitution by Cell

    Science.gov (United States)

    Greenhut, Stephanie; Jones, Megan

    2010-01-01

    On their visit to the National Archives Experience in Washington, D.C., students in Jenni Ashley and Gay Brock's U.S. history classes at the Potomac School in McLean, Virginia, participated in a pilot program called "The Constitution by Cell." Armed with their cell phones, a basic understanding of the Constitution, and a willingness to…

  14. Teaching About the Constitution.

    Science.gov (United States)

    White, Charles S.

    1988-01-01

    Reviews "The U.S. Constitution Then and Now," a two-unit program using the integrated database and word processing capabilities of AppleWorks. For grades 7-12, the units simulate the constitutional convention and the principles of free speech and privacy. Concludes that with adequate time, the program can provide a potentially powerful…

  15. The Constitution by Cell

    Science.gov (United States)

    Greenhut, Stephanie; Jones, Megan

    2010-01-01

    On their visit to the National Archives Experience in Washington, D.C., students in Jenni Ashley and Gay Brock's U.S. history classes at the Potomac School in McLean, Virginia, participated in a pilot program called "The Constitution by Cell." Armed with their cell phones, a basic understanding of the Constitution, and a willingness to…

  16. Evaluation of thermal resistance constitution for packaged AlGaN/GaN high electron mobility transistors by structure function method

    Institute of Scientific and Technical Information of China (English)

    Zhang Guang-Chen; Feng Shi-Wei; Zhou Zhou; Li Jing-Wan; Guo Chun-Sheng

    2011-01-01

    The evaluation of thermal resistance constitution for packaged AlGaN/GaN high electron mobility transistor (HEMT) by structure function method is proposed in this paper. The evaluation is based on the transient heating measurement of the AlGaN/GaN HEMT by pulsed electrical temperature sensitive parameter method. The extracted chip-level and package-level thermal resistances of the packaged multi-finger AlGaN/GaN HEMT with 400-μn SiC substrate are 22.5 K/W and 7.2 K/W respectively, which provides a non-invasive method to evaluate the chip-level thermal resistance of packaged AlGaN/GaN HEMTs. It is also experimentally proved that the extraction of the chip-level thermal resistance by this proposed method is not influenced by package form of the tested device and temperature boundary condition of measurement stage.

  17. Water deficits and heat shock effects on photosynthesis of a transgenic Arabidopsis thaliana constitutively expressing ABP9, a bZIP transcription factor

    DEFF Research Database (Denmark)

    Zhang, Xia; Wollenweber, Bernd; Jiang, Dong

    2008-01-01

    The effects of water deficits (WD), heat shock (HS), and both (HSWD) on photosynthetic carbon- and light-use efficiencies together with leaf ABA content, pigment composition and expressions of stress- and light harvesting-responsive genes were investigated in ABP9 [ABA-responsive-element (ABRE......, altered expression of stress-regulated or light harvesting-responsive genes was observed. Collectively, our results indicate that constitutive expression of ABP9 improves the photosynthetic capacity of plants under stress by adjusting photosynthetic pigment composition, dissipating excess light energy......) binding protein 9] transgenic Arabidopsis (5P2). WD, HS, and HSWD significantly decreased photosynthetic rate (A) and stomatal conductance (gs) in wild-type plants (WT). A and gs of 5P2 transgenic plants were slightly reduced by a single stress and were hardly modified by HSWD. Although A and electron...

  18. Protein cryoprotective activity of a cytosolic small heat shock protein that accumulates constitutively in chestnut stems and is up-regulated by low and high temperatures.

    Science.gov (United States)

    Lopez-Matas, Maria-Angeles; Nuñez, Paulina; Soto, Alvaro; Allona, Isabel; Casado, Rosa; Collada, Carmen; Guevara, Maria-Angeles; Aragoncillo, Cipriano; Gomez, Luis

    2004-04-01

    Heat shock, and other stresses that cause protein misfolding and aggregation, trigger the accumulation of heat shock proteins (HSPs) in virtually all organisms. Among the HSPs of higher plants, those belonging to the small HSP (sHSP) family remain the least characterized in functional terms. We analyzed the occurrence of sHSPs in vegetative organs of Castanea sativa (sweet chestnut), a temperate woody species that exhibits remarkable freezing tolerance. A constitutive sHSP subject to seasonal periodic changes of abundance was immunodetected in stems. This protein was identified by matrix-assisted laser-desorption ionization time of flight mass spectrometry and internal peptide sequencing as CsHSP17.5, a cytosolic class I sHSP previously described in cotyledons. Expression of the corresponding gene in stems was confirmed through cDNA cloning and reverse transcription-PCR. Stem protein and mRNA profiles indicated that CsHSP17.5 is significantly up-regulated in spring and fall, reaching maximal levels in late summer and, especially, in winter. In addition, cold exposure was found to quickly activate shsp gene expression in both stems and roots of chestnut seedlings kept in growth chambers. Our main finding is that purified CsHSP17.5 is very effective in protecting the cold-labile enzyme lactate dehydrogenase from freeze-induced inactivation (on a molar basis, CsHSP17.5 is about 400 times more effective as cryoprotectant than hen egg-white lysozyme). Consistent with these observations, repeated freezing/thawing did not affect appreciably the chaperone activity of diluted CsHSP17.5 nor its ability to form dodecameric complexes in vitro. Taken together, these results substantiate the hypothesis that sHSPs can play relevant roles in the acquisition of freezing tolerance.

  19. Right Product, Wrong Packaging: Not 'Constitution', but 'Constitutional Charter'

    Directory of Open Access Journals (Sweden)

    John Law

    2007-05-01

    Full Text Available The article seeks to locate the principal cause of Europe’s prevailing ratification crisis in the inappropriate title arrived at in the European Convention, Treaty Establishing a Constitution for Europe. This over-ambitious styling led the media to characterise the text as simply an ‘EU Constitution’. Yet, the text was not a Constitution as we traditionally understand the term, i.e. the founding document of a State: scholars are agreed that the EU is not, and will not become upon ratification, a State.In terms of substance, whilst the text certainly strengthened some emerging constitutional aspects, it was not a major departure from the status quo like the Single European Act and Treaty on European Union had been; and it remained technically a treaty like all its predecessors. Arguably, therefore, it did not require referenda to ratify. However, confusion over the scale and importance of what was proposed, stemming from ambiguity in the title, pushed politicians down this unfortunate path.The article identifies a high level of consensus among commentators as to the true nature of the text: most are happy designating it a treaty (noun with constitutional (adjective aspects. The early proposed title Constitutional Treaty for Europe was arguably, therefore, the correct one; but it is now too late to choose this option, as the terms Constitution and Constitutional Treaty have already been muddled in debate. A more distinctive change is required. One idea could be to follow the principle employed elsewhere in the text of codifying the generally accepted but presently unwritten legal concepts of the European Court of Justice, as was done for example for ‘primacy’ and ‘direct effect’. The Court has characterised the EU treaties as a ‘constitutional charter’ for over twenty years now, and on this basis a modified title could read Treaty Establishing a Constitutional Charter for Europe. Importantly, the term ‘charter’ is recognised

  20. Characterization of differentially expressed genes using high-dimensional co-expression networks

    DEFF Research Database (Denmark)

    Coelho Goncalves de Abreu, Gabriel; Labouriau, Rodrigo S.

    2010-01-01

    of spurious information along the network are avoided. The proposed inference procedure is based on the minimization of the Bayesian Information Criterion (BIC) in the class of decomposable graphical models. This class of models can be used to represent complex relationships and has suitable properties...... construct a compact representation of the co-expression network that allows to identify the regions with high concentration of differentially expressed genes. It is argued that differentially expressed genes located in highly interconnected regions of the co-expression network are less informative than...

  1. Constitutive SoxS expression in a fluoroquinolone-resistant strain with a truncated SoxR protein and identification of a new member of the marA-soxS-rob regulon, mdtG.

    Science.gov (United States)

    Fàbrega, Anna; Martin, Robert G; Rosner, Judah L; Tavio, M Mar; Vila, Jordi

    2010-03-01

    Elevated levels of fluoroquinolone resistance are frequently found among Escherichia coli clinical isolates. This study investigated the antibiotic resistance mechanisms of strain NorE5, derived in vitro by exposing an E. coli clinical isolate, PS5, to two selection steps with increasing concentrations of norfloxacin. In addition to the amino acid substitution in GyrA (S83L) present in PS5, NorE5 has an amino acid change in ParC (S80R). Furthermore, we now find by Western blotting that NorE5 has a multidrug resistance phenotype resulting from the overexpression of the antibiotic resistance efflux pump AcrAB-TolC. Microarray and gene fusion analyses revealed significantly increased expression in NorE5 of soxS, a transcriptional activator of acrAB and tolC. The high soxS activity is attributable to a frameshift mutation that truncates SoxR, rendering it a constitutive transcriptional activator of soxS. Furthermore, microarray and reverse transcription-PCR analyses showed that mdtG (yceE), encoding a putative efflux pump, is overexpressed in the resistant strain. SoxS, MarA, and Rob activated an mdtG::lacZ fusion, and SoxS was shown to bind to the mdtG promoter, showing that mdtG is a member of the marA-soxS-rob regulon. The mdtG marbox sequence is in the backward or class I orientation within the promoter, and its disruption resulted in a loss of inducibility by MarA, SoxS, and Rob. Thus, chromosomal mutations in parC and soxR are responsible for the increased antibiotic resistance of NorE5.

  2. Comparison of efficacy of the disease-specific LOX1- and constitutive cytomegalovirus-promoters in expressing interleukin 10 through adeno-associated virus 2/8 delivery in atherosclerotic mice.

    Directory of Open Access Journals (Sweden)

    Hongqing Zhu

    Full Text Available The development of gene therapy vectors for treating diseases of the cardiovascular system continues at a steady pace. Moreover, in the field of gene therapy the utility of "disease-specific promoters" has strong appeal. Many therapeutic genes, including transforming growth factor beta 1 or interleukin 10, are associated to adverse effects. The use of a disease-specific promoter might minimize toxicity. The lectin-like oxidized low density lipoprotein receptor 1 is a marker of cardiovascular disease and a potential therapeutic target. The lectin-like oxidized low density lipoprotein receptor 1 is known to be up-regulated early during disease onset in a number of cell types at the sites where the disease will be clinically evident. In this study an adeno-associated virus-2 DNA vector (AAV2 using the AAV8 capsid, and containing the full length The lectin-like oxidized low density lipoprotein receptor 1 promoter, was generated and assayed for its ability to express human interleukin 10 in low density lipoprotein receptor knockout mice on high cholesterol diet. The cytomegalovirus early promoter was used for comparison in a similarly structured vector. The two promoters were found to have equal efficacy in reducing atherogenesis as measured by aortic systolic blood velocity, aortic cross sectional area, and aortic wall thickness. This is the first head-to-head comparison of a constitutive with a disease-specific promoter in a therapeutic context. These data strongly suggest that the use of a disease-specific promoter is appropriate for therapeutic gene delivery.

  3. Rat mammary carcinogenesis induced by in situ expression of constitutive Raf kinase activity is prevented by tethering Raf to the plasma membrane.

    Science.gov (United States)

    McFarlin, Daniel R; Gould, Michael N

    2003-06-01

    Mammary carcinogenesis induced through expression of activated Raf was investigated using a model in which retroviral vectors were infused into the central ducts of rat mammary glands. This model allows efficient expression of experimental proteins in a small fraction of endogenous mammary epithelial cells in situ. We previously reported that Raf is the dominant oncogenic signaling pathway from activated Ras in rat mammary glands. We show here that mammary gland carcinogenesis is rapidly induced by the expression of c-Raf-1 kinase that is activated by N-terminal truncation (Delta-Raf). Interestingly, targeting Raf to the plasma membrane via C-terminal fusion with Ras membrane localization signals (Raf-Caax) induces Raf kinase activity that transforms 3T3 cells more frequently than Delta-Raf, yet in situ expression of Raf-Caax does not induce mammary carcinomas. To investigate these contrasting results and begin elucidating the mechanisms of Raf-induced mammary carcinogenesis, we combined both activating mutations (N-terminal truncation and C-terminal membrane localization motifs) in one Raf construct (Delta-Raf-Caax). While Delta-Raf-Caax transforms 3T3 cells more efficiently than Delta-Raf or Raf-Caax, in situ expression of Delta-Raf-Caax does not induce carcinomas in vivo, demonstrating that lipid modification on the C-terminus of Delta-Raf negates its oncogenic potential in rat mammary gland.

  4. A Venture in Constitutional Law.

    Science.gov (United States)

    Cole, W. Graham; Dillon, Dorothy H.

    1980-01-01

    Senior high girls and boys from two single-sex schools undertook a study of a Supreme Court case that provided insight not only into constitutional law and history but also into how men and women can work together and relate in other ways than dating. (DS)

  5. Efficiency of the technique of the training process of highly skilled bodybuilders of the mesomorphic type of constitution in the competitive period

    Directory of Open Access Journals (Sweden)

    Olexandr Tyhorskіy

    2016-10-01

    Full Text Available Purpose: foundation of efficiency of the technique of the training process of highly skilled bodybuilders in the competitive period. Material & Methods: 16 highly skilled bodybuilders of 22–30 years old participated in the research, the average body weight of sportsmen makes 872 – 1022 kg, which are included in the structure of the national team of Ukraine on bodybuilding. Methods: method of the theoretical analysis and generalization of literature, pedagogical observation, pedagogical experiment, method of mathematical statistics. Results: the comparative characteristic of the most often used techniques of the training process in bodybuilding is provided. The effective technique for highly skilled bodybuilders of the mesomorphic type of constitution, depending on the initial uniform of a sportsman at the beginning of the competitive period of training is developed and proved. Dependence of change of body weight of a bodybuilder on the training process is directed. Conclusions: on the effective training method, depending on microcycle of trainings in the competitive period precompetitive and competitive mesocycles (selection and main competitions, is offered the basis of the conducted research.

  6. Constitution, 29 October 1987.

    Science.gov (United States)

    1987-01-01

    This document contains provisions of chapter 2 (Rights and Duties of Citizens) of the 1987 Constitution of the Republic of Korea relating to equality of the sexes, freedom of movement, free and compulsory education, equal opportunity at work, social protection, housing, and health care. The Constitution states that all citizens are equal before the law with no discrimination in political, economic, social, or cultural life based on sex, religion, or social status. The Constitution also protects freedom of residence and mobility, the right to an equal education, free compulsory education, and working mothers (with specific protection against discrimination). The Constitution directs the state to promote the welfare and rights of women, enhance the welfare of the aged and the young, and protect those incapable of earning a living. Housing development policies will be used to ensure comfortable housing for all citizens. State protection is afforded to mothers and to the health of all citizens.

  7. High-level expression, purification, polyclonal antibody preparation ...

    African Journals Online (AJOL)

    user

    2011-02-14

    Feb 14, 2011 ... Full Length Research Paper. High-level expression ... resistance severely compromises effective therapeutic options. ... In the present study, we first report the expression of the oprD ... databases of National Center for Biotechnology Information (NCBI) ..... assembly of the head of bacteriophage T4. Nature.

  8. Constitutive expression of interferon-induced human MxA protein in transgenic tobacco plants does not confer resistance to a variety of RNA viruses

    NARCIS (Netherlands)

    Frese, M.; Prins, M.; Ponten, A.; Goldbach, R.W.; Haller, O.; Zeltz, P.

    2000-01-01

    MxA is a key component in the interferon-induced antiviral defense in humans. After viral infections, MxA is rapidly induced and accumulates in the cytoplasm. The multiplication of many RNA viruses,including all bunyaviruses tested so far, is inhibited by MxA. These findings prompted us to express

  9. Constitutive expression of interferon-induced human MxA protein in transgenic tobacco plants does not confer resistance to a variety of RNA viruses

    NARCIS (Netherlands)

    Frese, M.; Prins, M.; Ponten, A.; Goldbach, R.W.; Haller, O.; Zeltz, P.

    2000-01-01

    MxA is a key component in the interferon-induced antiviral defense in humans. After viral infections, MxA is rapidly induced and accumulates in the cytoplasm. The multiplication of many RNA viruses,including all bunyaviruses tested so far, is inhibited by MxA. These findings prompted us to express M

  10. Hateful Expression and First Amendment Values: Toward a Theory of Constitutional Constraint on Hate Speech at Colleges and Universities after R.A.V. v. St. Paul.

    Science.gov (United States)

    Hartman, Rhonda G.

    1993-01-01

    Discussion of hate speech on college campuses focuses on whether the concept of hate speech encompasses expression deserving full First Amendment protection and thus virtual immunity from college regulation. It is argued that the Supreme Court could sustain some form of hate-speech policy on campus based on precedent elsewhere. (MSE)

  11. A macro-mechanical constitutive model for shape memory polymer

    Institute of Scientific and Technical Information of China (English)

    2010-01-01

    It is of theoretical and engineering interest to establish a macro-mechanical constitutive model of the shape memory polymer (SMP), which includes the mechanical constitutive equation and the material parameter function, from the viewpoint of practical application. In this paper, a new three-dimensional macro-mechanical constitutive equation, which describes the mechanical behaviors associated with the shape memory effect of SMP, is developed based on solid mechanics and the viscoelasticity theorem. According to the results of the DMA test, a new material parameter function is established to express the relationship of the material parameters and temperature during the glass transition of SMP. The new macro-mechanical constitutive equation and material parameter function are used to numerically simulate the process producing the shape memory effect of SMP, which includes deforming at high temperature, stress freezing, unloading at low temperature and shape recovery. They are also used to investigate and analyze the influences of loading rate and temperature change rate on the thermo-mechanical behaviors of SMP. The numerical results and the comparisons with Zhou’s material parameter function and Tobushi’s mechanical constitutive equation illustrate that the proposed three-dimensional macro-mechanical constitutive model can effectively predict the thermo-mechanical behaviors of SMP under the state of complex stress.

  12. Isolation and characterization of "GmScream" promoters that regulate highly expressing soybean (Glycine max Merr.) genes.

    Science.gov (United States)

    Zhang, Ning; McHale, Leah K; Finer, John J

    2015-12-01

    To increase our understanding of the regulatory components that control gene expression, it is important to identify, isolate and characterize new promoters. In this study, a group of highly expressed soybean (Glycine max Merr.) genes, which we have named "GmScream", were first identified from RNA-Seq data. The promoter regions were then identified, cloned and fused with the coding region of the green fluorescent protein (gfp) gene, for introduction and analysis in different tissues using 3 tools for validation. Approximately half of the GmScream promoters identified showed levels of GFP expression comparable to or higher than the Cauliflower Mosaic Virus 35S (35S) promoter. Using transient expression in lima bean cotyledonary tissues, the strongest GmScream promoters gave over 6-fold higher expression than the 35S promoter while several other GmScream promoters showed 2- to 3-fold higher expression. The two highest expressing promoters, GmScreamM4 and GmScreamM8, regulated two different elongation factor 1A genes in soybean. In stably transformed soybean tissues, GFP driven by the GmScreamM4 or GmScreamM8 promoter exhibited constitutive high expression in most tissues with preferentially higher expression in proliferative embryogenic tissues, procambium, vascular tissues, root tips and young embryos. Using deletion analysis of the promoter, two proximal regions of the GmScreamM8 promoter were identified as contributing significantly to high levels of gene expression.

  13. A Novel Mutation within the Central Listeria monocytogenes Regulator PrfA That Results in Constitutive Expression of Virulence Gene Products

    OpenAIRE

    Wong, Kendy K. Y.; Freitag, Nancy E.

    2004-01-01

    The PrfA protein of Listeria monocytogenes functions as a key regulatory factor for the coordinated expression of many virulence genes during bacterial infection of host cells. PrfA activity is controlled by multiple regulatory mechanisms, including an apparent requirement for either the presence of a cofactor or some form of posttranslational modification that regulates the activation of PrfA. In this study, we describe the identification and characterization of a novel PrfA mutation that re...

  14. Multi-colony stimulating activity of interleukin 5 (IL-5) on hematopoietic progenitors from transgenic mice that express IL-5 receptor alpha subunit constitutively

    OpenAIRE

    1995-01-01

    The interleukin 3 (IL-3), IL-5, and granulocyte/macrophage colony- stimulating factor receptors consist of a cytokine-specific alpha subunit and the common beta subunit. Whereas IL-3 stimulates various lineages of hematopoietic cells, including multipotential progenitors, IL-5 acts mainly as an eosinophil lineage-specific factor. To investigate whether the lineage specificity of IL-5 is due to restricted expression of the IL-5 receptor alpha subunit (IL-5R alpha), we generated transgenic mice...

  15. Constitutive receptor systems for drug discovery.

    Science.gov (United States)

    Chen, G; Jayawickreme, C; Way, J; Armour, S; Queen, K; Watson, C; Ignar, D; Chen, W J; Kenakin, T

    1999-12-01

    This paper discusses the use of constitutively active G-protein-coupled receptor systems for drug discovery. Specifically, the ternary complex model is used to define the two major theoretical advantages of constitutive receptor screening-namely, the ability to detect antagonists as well as agonists directly and the fact that constitutive systems are more sensitive to agonists. In experimental studies, transient transfection of Chinese hamster ovary cyclic AMP response element (CRE) luciferase reporter cells with cDNA for human parathyroid hormone receptor, glucagon receptor, and glucagon-like peptide (GLP-1) receptor showed cDNA concentration-dependent constitutive activity with parathyroid hormone (PTH-1) and glucagon. In contrast, no constitutive activity was observed for GLP-1 receptor, yet responses to GLP-1 indicated that receptor expression had taken place. In another functional system, Xenopus laevi melanophores transfected with cDNA for human calcitonin receptor showed constitutive activity. Nine ligands for the calcitonin receptor either increased or decreased constitutive activity in this assay. The sensitivity of the system to human calcitonin increased with increasing constitutive activity. These data indicate that, for those receptors which naturally produce constitutive activity, screening in this mode could be advantageous over other methods.

  16. Mathematical Modeling of High-Temperature Constitutive Equations and Hot Processing Maps for As-Cast SA508-3 Steel

    Science.gov (United States)

    Sui, Dashan; Wang, Tao; Zhu, Lingling; Gao, Liang; Cui, Zhenshan

    2016-08-01

    The hot deformation behavior and hot workability characteristics of as-cast SA508-3 steel were studied by modeling the constitutive equations and developing hot processing maps. The isothermal compression experiments were carried out at temperatures of 950°C, 1050°C, 1150°C, and 1250°C and strain rates of 0.001 s-1, 0.01 s-1, 0.1 s-1, and 1 s-1 respectively. The two-stage flow stress models were established through the classical theories on work hardening and softening, and the solution of activation energy for hot deformation was 355.0 kJ mol-1 K-1. Based on the dynamic material model, the power dissipation and instability maps were developed separately at strains of 0.2, 0.4, 0.6 and 0.8. The power dissipation rate increases with both the increase of temperature and the decrease of strain rate, and the instable region mainly appears on the conditions of low temperature and high strain rate. The optimal hot working parameters for as-cast SA508-3 steel are 1050-1200°C/0.001-0.1 s-1, with about 25-40% peak efficiency of power dissipation.

  17. A high constitutive catalase activity confers resistance to methyl viologen-promoted oxidative stress in a mutant of the cyanobacterium Nostoc punctiforme ATCC 29133.

    Science.gov (United States)

    Moirangthem, Lakshmipyari Devi; Bhattacharya, Sudeshna; Stensjö, Karin; Lindblad, Peter; Bhattacharya, Jyotirmoy

    2014-04-01

    A spontaneous methyl viologen (MV)-resistant mutant of the nitrogen-fixing cyanobacterium Nostoc punctiforme ATCC 29133 was isolated and the major enzymatic antioxidants involved in combating MV-induced oxidative stress were evaluated. The mutant displayed a high constitutive catalase activity as a consequence of which, the intracellular level of reactive oxygen species in the mutant was lower than the wild type (N. punctiforme) in the presence of MV. The superoxide dismutase (SOD) activity that consisted of a SodA (manganese-SOD) and a SodB (iron-SOD) was not suppressed in the mutant following MV treatment. The mutant was, however, characterised by a lower peroxidase activity compared with its wild type, and its improved tolerance to externally added H₂O₂ could only be attributed to enhanced catalase activity. Furthermore, MV-induced toxic effects on the wild type such as (1) loss of photosynthetic performance assessed as maximal quantum yield of photosystem II, (2) nitrogenase inactivation, and (3) filament fragmentation and cell lysis were not observed in the mutant. These findings highlight the importance of catalase in preventing MV-promoted oxidative damage and cell death in the cyanobacterium N. punctiforme. Such oxidative stress resistant mutants of cyanobacteria are likely to be a better source of biofertilisers, as they can grow and fix nitrogen in an unhindered manner in agricultural fields that are often contaminated with the herbicide MV, also commonly known as paraquat.

  18. Beyond the Economic Constitution

    DEFF Research Database (Denmark)

    Kjær, Poul F.

    -twentieth century. In particular, the “turn to corporatism” in the interwar period implied an erosion of the fragile institutionalisation of legally-constituted public power due to its suspension of the legal infrastructure of society and the concomitant breakdown of the distinction between the public and private...... realms of society. The (trans-) national re-constitution of Western Europe in the years immediately after WWII, which the European integration process was an integrated part of, successfully remedied this development. However, over the last decades, Europe, together with the rest of the world, has...

  19. Immunoinformatics study on highly expressed Mycobacterium tuberculosis genes during infection.

    Science.gov (United States)

    Nguyen Thi, Le Thuy; Sarmiento, Maria Elena; Calero, Romel; Camacho, Frank; Reyes, Fatima; Hossain, Md Murad; Gonzalez, Gustavo Sierra; Norazmi, Mohd Nor; Acosta, Armando

    2014-09-01

    The most important targets for vaccine development are the proteins that are highly expressed by the microorganisms during infection in-vivo. A number of Mycobacterium tuberculosis (Mtb) proteins are also reported to be expressed in-vivo at different phases of infection. In the present study, we analyzed multiple published databases of gene expression profiles of Mtb in-vivo at different phases of infection in animals and humans and selected 38 proteins that are highly expressed in the active, latent and reactivation phases. We predicted T- and B-cell epitopes from the selected proteins using HLAPred for T-cell epitope prediction and BCEPred combined with ABCPred for B-cell epitope prediction. For each selected proteins, regions containing both T- and B-cell epitopes were identified which might be considered as important candidates for vaccine design against tuberculosis.

  20. Regulated expression of CXCR4 constitutive active mutants revealed the up-modulated chemotaxis and up-regulation of genes crucial for CXCR4 mediated homing and engraftment of hematopoietic stem/progenitor cells

    Directory of Open Access Journals (Sweden)

    Sharma M

    2013-04-01

    Full Text Available SDF-1/CXCR4 axis plays a principle role in the homing and engraftment of hematopoietic stem/progenitor cells (HSPCs, a process that defines cells ability to reach and seed recipient bone marrow niche following their intravenous infusion. However, the proper functioning of CXCR4 downstream signaling depends upon consistent optimal expression of both SDF-1 ligand and its receptor CXCR4, which in turn is variable and regulated by several factors. The constitutive active mutants of CXCR4 (N119A and N119S being able to induce autonomous downstream signaling, overcome the limitation of ligand-receptor interaction for induction of CXCR4 signaling. Therefore, we intended to explore their potential in chemotaxis; a key cellular process which crucially regulates cells homing to bone marrow. In present study, Tet-on inducible gene expression vector system was used for doxycycline inducible regulated transgene expression of CXCR4 active mutants in hematopoietic stem progenitor cell line K-562. Both of these mutants revealed significantly enhanced chemotaxis to SDF-1 gradient as compared to wild type. Furthermore, gene expression profiling of these genetically engineered cells as assessed by microarray analysis revealed the up-regulation of group of genes that are known to play a crucial role in CXCR4 mediated cells homing and engraftment. Hence, this study suggest the potential prospects of CXCR4 active mutants in research and development aimed to improve the efficiency of cells in the mechanism of homing and engraftment process.

  1. Crushed Salt Constitutive Model

    Energy Technology Data Exchange (ETDEWEB)

    Callahan, G.D.

    1999-02-01

    The constitutive model used to describe the deformation of crushed salt is presented in this report. Two mechanisms -- dislocation creep and grain boundary diffusional pressure solution -- are combined to form the basis for the constitutive model governing the deformation of crushed salt. The constitutive model is generalized to represent three-dimensional states of stress. Upon complete consolidation, the crushed-salt model reproduces the Multimechanism Deformation (M-D) model typically used for the Waste Isolation Pilot Plant (WIPP) host geological formation salt. New shear consolidation tests are combined with an existing database that includes hydrostatic consolidation and shear consolidation tests conducted on WIPP and southeastern New Mexico salt. Nonlinear least-squares model fitting to the database produced two sets of material parameter values for the model -- one for the shear consolidation tests and one for a combination of the shear and hydrostatic consolidation tests. Using the parameter values determined from the fitted database, the constitutive model is validated against constant strain-rate tests. Shaft seal problems are analyzed to demonstrate model-predicted consolidation of the shaft seal crushed-salt component. Based on the fitting statistics, the ability of the model to predict the test data, and the ability of the model to predict load paths and test data outside of the fitted database, the model appears to capture the creep consolidation behavior of crushed salt reasonably well.

  2. Sexuality and the Constitution.

    Science.gov (United States)

    Copelon, Rhonda

    1987-01-01

    Argues for abortion rights and protection of intimate decisions and relationships. Describes the role and position of women in eighteenth century American society as a means of exposing the fallacy of the anti-abortion movement's insistence on adherence to constitutional text. Discusses the recent attempts to overturn the Roe v. Wade ruling. (PS)

  3. The Constitutional Heritage.

    Science.gov (United States)

    Baxter, Maurice

    Changing political, social, economic, and intellectual conditions over the past two hundred years have demanded innovation and adjustment of legal doctrine, thus giving the United States Constitution a character which the framers of the document could not have predicted. Historically, one must not only understand developments since 1787 but also…

  4. Constitutional Law--Elective.

    Science.gov (United States)

    Gallagher, Joan; Wood, Robert J.

    The elective unit on Constitutional Law is intended for 11th and 12th grade students. The unit is designed around major course goals which are to develop those concepts whereby students recognize and understand the following three topic areas: 1) Role of the Federal Judicial Branch of Government, 2) Supreme Court Cases Involving the Three Branches…

  5. Communicative Constitution of Organizations

    DEFF Research Database (Denmark)

    Schoeneborn, Dennis; Vasquez, Consuelo

    2017-01-01

    The notion of the communicative constitution of organizations (CCO) is at the center of a growing theoretical development within organizational communication studies. CCO scholarship is based on the idea that organization emerges in and is sustained and transformed by communication. This entry pr...

  6. ~Ylectoral Systems, Constitutionalism

    African Journals Online (AJOL)

    strongly that for democratic governance and constitutionalism to be nurtured and consolidated, SADC states need to 'undergo deliberate electoral system ...... either nmke or hrez1l< the seen1ing_|y emergent tW0—pm"ty ((t|.1opoiy) system in.

  7. The Constitution in Action

    Science.gov (United States)

    Potter, Lee Ann

    2007-01-01

    In this article, the author describes the experiences middle school students on a field trip to the new Constitution in Action Learning Lab in the Boeing Learning Center at the National Archives can expect. There, middle school students take on the roles of archivists and researchers collecting and analyzing primary sources from the holdings of…

  8. The Constitution of Partnering

    DEFF Research Database (Denmark)

    Gottlieb, Stefan Christoffer

    The constitution of partnering. Afhandlingen behandler konstitueringen af ledelseskonceptet partnering og dets anvendelse i dansk byggeri. Partnering er et udbredt koncept i byggeriet som betoner samarbejde, tillid og gensidighed mellem de deltagende parter, og konceptet har de senere år har været...

  9. Rapid transcriptional pulsing dynamics of high expressing retroviral transgenes in embryonic stem cells.

    Directory of Open Access Journals (Sweden)

    Mandy Y M Lo

    Full Text Available Single cell imaging studies suggest that transcription is not continuous and occurs as discrete pulses of gene activity. To study mechanisms by which retroviral transgenes can transcribe to high levels, we used the MS2 system to visualize transcriptional dynamics of high expressing proviral integration sites in embryonic stem (ES cells. We established two ES cell lines each bearing a single copy, self-inactivating retroviral vector with a strong ubiquitous human EF1α gene promoter directing expression of mRFP fused to an MS2-stem-loop array. Transfection of MS2-EGFP generated EGFP focal dots bound to the mRFP-MS2 stem loop mRNA. These transcription foci colocalized with the transgene integration site detected by immunoFISH. Live tracking of single cells for 20 minutes detected EGFP focal dots that displayed frequent and rapid fluctuations in transcription over periods as short as 25 seconds. Similarly rapid fluctuations were detected from focal doublet signals that colocalized with replicated proviral integration sites by immunoFISH, consistent with transcriptional pulses from sister chromatids. We concluded that retroviral transgenes experience rapid transcriptional pulses in clonal ES cell lines that exhibit high level expression. These events are directed by a constitutive housekeeping gene promoter and may provide precedence for rapid transcriptional pulsing at endogenous genes in mammalian stem cells.

  10. Statistical Constitutive Equation of Aramid Fiber Bundles

    Institute of Scientific and Technical Information of China (English)

    熊杰; 顾伯洪; 王善元

    2003-01-01

    Tensile impact tests of aramid (Twaron) fiber bundles were carried om under high strain rates with a wide range of 0. 01/s~1000/s by using MTS and bar-bar tensile impact apparatus. Based on the statistical constitutive model of fiber bundles, statistical constitutive equations of aramid fiber bundles are derived from statistical analysis of test data at different strain rates. Comparison between the theoretical predictions and experimental data indicates statistical constitutive equations fit well with the experimental data, and statistical constitutive equations of fiber bundles at different strain rates are valid.

  11. In vitro metacyclogenesis of Trypanosoma cruzi induced by starvation correlates with a transient adenylyl cyclase stimulation as well as with a constitutive upregulation of adenylyl cyclase expression.

    Science.gov (United States)

    Hamedi, Afsaneh; Botelho, Larisse; Britto, Constança; Fragoso, Stenio Perdigão; Umaki, Adriana Castilhos Souza; Goldenberg, Samuel; Bottu, Guy; Salmon, Didier

    2015-01-01

    The Trypanosoma cruzi adenylyl cyclase (AC) multigene family encodes different isoforms (around 15) sharing a variable large N-terminal domain, which is extracellular and receptor-like, followed by a transmembrane helix and a conserved C-terminal catalytic domain. It was proposed that these key enzymes in the cAMP signalling pathway allow the parasite to sense its changing extracellular milieu in order to rapidly adapt to its new environment, which is generally achieved through a differentiation process. One of the critical differentiation events the parasitic protozoan T. cruzi undergoes during its life cycle, known as metacyclogenesis, occurs in the digestive tract of the insect and corresponds to the differentiation from noninfective epimastigotes to infective metacyclic trypomastigote forms. By in vitro monitoring the activity of AC during metacyclogenesis, we showed that both the activity of AC and the intracellular cAMP content follow a similar pattern of transient stimulation in a two-step process, with a first activation peak occurring during the first hours of nutritional stress and a second peak between 6 and 48 h, corresponding to the cellular adhesion. During this differentiation process, a general mechanism of upregulation of AC expression of both mRNA and protein is triggered and in particular for a major subclass of these enzymes that are present in various gene copies commonly associated to the THT gene clusters. Although the scattered genome distribution of these gene copies is rather unusual in trypanosomatids and seems to be a recent acquisition in the evolution of the T. cruzi clade, their encoded product redistributed on the flagellum of the parasite upon differentiation could be important to sense the extracellular milieu.

  12. What constitutes information integrity?

    Directory of Open Access Journals (Sweden)

    S. Flowerday

    2007-12-01

    Full Text Available This research focused on what constitutes information integrity as this is a problem facing companies today. Moreover, information integrity is a pillar of information security and is required in order to have a sound security management programme. However, it is acknowledged that 100% information integrity is not currently achievable due to various limitations and therefore the auditing concept of reasonable assurance is adopted. This is in line with the concept that 100% information security is not achievable and the notion that adequate security is the goal, using appropriate countermeasures. The main contribution of this article is to illustrate the importance of and provide a macro view of what constitutes information integrity. The findings are in harmony with Samuel Johnson's words (1751: 'Integrity without knowledge is weak and useless, and knowledge without integrity is dangerous and dreadful.'

  13. What constitutes information integrity?

    Directory of Open Access Journals (Sweden)

    S. Flowerday

    2008-01-01

    Full Text Available This research focused on what constitutes information integrity as this is a problem facing companies today. Moreover, information integrity is a pillar of information security and is required in order to have a sound security management programme. However, it is acknowledged that 100% information integrity is not currently achievable due to various limitations and therefore the auditing concept of reasonable assurance is adopted. This is in line with the concept that 100% information security is not achievable and the notion that adequate security is the goal, using appropriate countermeasures. The main contribution of this article is to illustrate the importance of and provide a macro view of what constitutes information integrity. The findings are in harmony with Samuel Johnson's words (1751: 'Integrity without knowledge is weak and useless, and knowledge without integrity is dangerous and dreadful.'

  14. The Constitution in Other Lands.

    Science.gov (United States)

    Bill of Rights in Action, 1987

    1987-01-01

    Designed for classroom teaching, this document contains articles on the new constitutions of Japan, South Korea, and the Philippine Islands which were modeled in part on the U.S. Constitution. These countries' experiences with constitutional government are examined, and whether or not the U.S. Constitution can be a suitable model for other…

  15. 肾虚质大鼠学习记忆与BDNF表达的实验研究%Experimental study about learning and memory capacity and BDNF expression of rats with kidney-deficiency constitution

    Institute of Scientific and Technical Information of China (English)

    孙理军; 孙耀光; 王晓棣; 王兴; 孙瑜嬬; 张琪

    2015-01-01

    Objective:Through learning and memory capacity and hippocampus BDNF expression of rats with kidney‐deficiency constitution ,reveal the material basis and neurobiological mechanisms of kidney deficiency constitution .Methods:By the way of“cat scare rat” ,built composite offspring rat models with deficiency and acquired dystrophy ,then divided them into model group ,ZuoGui pill group and YouGui pill group ,and blank group offspring came from normal pregnant rats .After the treatment with kidney‐supplementing drugs ,learning and memory capacity was detected by Morris water maze test ,expressions of BDNF ,TrkB ,CaMK2 and CREB were detected by western blot method .Results:Morris water maze test :The incubation period ,total distance and the first time through target area of model group were longer than blank group ,while in ZuoGui pill and YouGui pill groups ,these indexes were lower than model group .BDNF expression of model group more declined than blank group .After treatment with kidney‐supplementing drugs ,BDNF expression was higher than model group .Conclusion:Rats with kidney‐deficiency constitution ,its learning and memory capacity and BDNF expression decline as well as TrKB ,CaMK2 and CREB .Kidney‐supplementing drugs ZuoGui pill and YouGui pill can increase all of them thereby improve the brain function deg‐radation .%目的:通过肾虚质大鼠学习记忆与海马BD N F及其相关分子的表达变化,揭示肾藏志理论的物质基础与神经生物学机制。方法:采用“猫吓鼠”经典造模法造成先天不足加后天失养复合型肾虚质模型仔鼠,分为模型组、左归丸、右归丸组,空白组产自正常孕鼠。选用补肾中药进行调理,Morris水迷宫实验观察学习记忆功能变化,免疫印迹法检测肾虚质大鼠海马脑源性神经营养因子(BDNF)、酪氨酸激酶B(TrkB )、钙/钙调素依赖蛋白激酶2(CaMK2)和cAMP反应原件结合蛋白(CREB)表达。

  16. Constitutive modeling for isotropic materials

    Science.gov (United States)

    Chan, K. S.; Lindholm, U. S.; Bodner, S. R.

    1988-01-01

    The third and fourth years of a 4-year research program, part of the NASA HOST Program, are described. The program goals were: (1) to develop and validate unified constitutive models for isotropic materials, and (2) to demonstrate their usefulness for structural analysis of hot section components of gas turbine engines. The unified models selected for development and evaluation were those of Bodner-Partom and of Walker. The unified approach for elastic-viscoplastic constitutive equations is a viable method for representing and predicting material response characteristics in the range where strain rate and temperature dependent inelastic deformations are experienced. This conclusion is reached by extensive comparison of model calculations against the experimental results of a test program of two high temperature Ni-base alloys, B1900+Hf and Mar-M247, over a wide temperature range for a variety of deformation and thermal histories including uniaxial, multiaxial, and thermomechanical loading paths. The applicability of the Bodner-Partom and the Walker models for structural applications has been demonstrated by implementing these models into the MARC finite element code and by performing a number of analyses including thermomechanical histories on components of hot sections of gas turbine engines and benchmark notch tensile specimens. The results of the 4-year program have been published in four annual reports. The results of the base program are summarized in this report. The tasks covered include: (1) development of material test procedures, (2) thermal history effects, and (3) verification of the constitutive model for an alternative material.

  17. 用GAP启动子在毕节酵母中组成型表达人血管抑制素%Constitutive Expression of Human Angiostatin in Pichia pastoris Using the GAP Promoter

    Institute of Scientific and Technical Information of China (English)

    张爱联; 罗进贤; 张添元; 陈守才; 官文俊

    2004-01-01

    The GAP gene promoter was amplified from P. pastoris GS115 and used to replace the AOX1 promoter( PAox1 )on pPIC9K resulting in plasmid pGAP9K. The recombinant expression vector pGAP9K-AS was constructed by inserting the angiostatin gene(AS) into pGAP9K, pGAP9K-AS was then transformed into P. pastoris GS115. The multi-copy integration transformant P. pastoris GS115 (pGAP9K-AS) was used to investigate the constitutive expression of angiostatin in P. pasto-ris. The expression of angiostatin reached its peak after 4 d of culture in P. pastoris GS115 (pGAP9K-AS) while the an-giostatin expressed in P. pastoris GS115 (pPIC9K-AS) after 4 d of induction or 5 d of culture is only 70% of that expressed by P. pastoris GS115(pGAP9K-AS). The AS expression in inducible system reached the peak after 6 d of induction but the expressed AS was only 86% of that from constitutive system. The results of anti-angiogenic and antitumor activity assay showed that AS expressed from both constitutive and inducible system inhibited the CAM angiogenesis and suppressed B16 melanoma in C57BL/6J mouse and that the tumor inhibition rates reached 90. 63% and 90. 54% ,respectively. The above data indicates that the constitutive promoter PGAP can served as an effective alternative to the inductive promoter PAox1 to ex-press AS and other proteins in P. pastoris.%为探索用GAP启动子(PGAP)取代AOX1启动子(PAOX1),在毕节酵母(P.pastoris)中组成型表达外源蛋白的可能性,应用PCR方法从P.pastoris染色体中扩增了GAP启动子,以其取代诱导型表达载体pPIC9K上的PAOX1,构建了组成型表达载体pGAP9K.将人血管抑制素(AS)基因重组于pGAP9K的多克隆位点,获得含AS基因的重组质粒pGAP9K-AS.转化P.pastoris GS115,对获得的高拷贝转化子P.pastoris GS115(pGAP9K-AS)进行组成型表达,同时以诱导型转化子P.pastoris GS115(pPIC9K-AS)作为对照.SDS-PAGE结果显示:组成型转化子于培养4 d后AS的表达水平

  18. Transnational Governance and Constitutionalism

    DEFF Research Database (Denmark)

    Joerges, Christian; Sand, Inger-Johanne; Teubner, Gunther

    The term transnational governance designates untraditional types of international and regional collaboration among both public and private actors. These legally-structured or less formal arrangements link economic, scientific and technological spheres with political and legal processes......, supranational nor totally denationalised. It is neither arbitrary nor accidental that we present our inquiries into this phenomenon in the series of International Studies on Private Law Theory....... of democratic governance. The book refers to this term as a yardstick to which then contributors feel committed even where they plead for a reconceptualisation of constitutionalism or a discussion of its functional equivalents. 'Transnational governance' is neither public nor private, nor purely international...

  19. Estimation of Nonlinear Three-dimensional Constitutive Law for DNA Molecules

    CERN Document Server

    Palanthandalam-Madapusi, Harish J

    2010-01-01

    Long length-scale structural deformations of DNA play a central role in many biological processes including gene expression. The elastic rod model, which uses a continuum approximation, has emerged as a viable tool to model deformations of DNA molecules. The elastic rod model predictions are however very sensitive to the constitutive law (material properties) of the molecule, which in turn, vary along the molecules length according to its base-pair sequence. Identification of the nonlinear sequence-dependent constitutive law from experimental data and feasible molecular dynamics simulations remains a significant challenge. In this paper, we develop techniques to use elastic rod model equations in combination with limited experimental measurements or high-fidelity molecular dynamics simulation data to estimate the nonlinear constitutive law governing DNA molecules. We first cast the elastic rod model equations in state-space form and express the effect of the unknown constitutive law as an unknown input to the...

  20. High lib mRNA expression in breast carcinomas.

    Science.gov (United States)

    Satoh, Kazuki; Hata, Mitsumi; Yokota, Hiroshi

    2004-06-30

    Lib, first identified as a novel beta-amyloid responsive gene in rat astrocytes, has an extracellular domain of 15 leucine-rich repeats (LRRs) followed by a transmembrane domain and a short cytoplasmic region. It is a distinctly inducible gene and is thought to play a key role in inflammatory states via the LRR extracellular motif, an ideal structural framework for protein-protein and protein-matrix interactions. To evaluate potential roles of Lib, we screened various tumors for Lib expression. Lib mRNA expression was high and uniquely expressed in breast tumor tissues, compared to paired normal breast tissues. Lib mRNA was localized in the ductal carcinoma cells and Lib protein displayed a homophilic association on the surface of cultured cells. These data suggest that Lib may play a role in the progression of breast carcinomas and may be a diagnostic marker for breast tumors.

  1. Computational Analysis and Low-Scale Constitutive Expression of Laccases Synthetic Genes GlLCC1 from Ganoderma lucidum and POXA 1B from Pleurotus ostreatus in Pichia pastoris

    Science.gov (United States)

    Reyes-Guzmán, Edwin Alfredo; Poutou-Piñales, Raúl A.; Reyes-Montaño, Edgar Antonio; Pedroza-Rodríguez, Aura Marina; Rodríguez-Vázquez, Refugio; Cardozo-Bernal, Ángela M.

    2015-01-01

    Lacasses are multicopper oxidases that can catalyze aromatic and non-aromatic compounds concomitantly with reduction of molecular oxygen to water. Fungal laccases have generated a growing interest due to their biotechnological potential applications, such as lignocellulosic material delignification, biopulping and biobleaching, wastewater treatment, and transformation of toxic organic pollutants. In this work we selected fungal genes encoding for laccase enzymes GlLCC1 in Ganoderma lucidum and POXA 1B in Pleurotus ostreatus. These genes were optimized for codon use, GC content, and regions generating secondary structures. Laccase proposed computational models, and their interaction with ABTS [2, 2′-azino-bis(3-ethylbenzothiazoline-6-sulphonic acid)] substrate was evaluated by molecular docking. Synthetic genes were cloned under the control of Pichia pastoris glyceraldehyde-3-phosphate dehydrogenase (GAP) constitutive promoter. P. pastoris X-33 was transformed with pGAPZαA-LaccGluc-Stop and pGAPZαA-LaccPost-Stop constructs. Optimization reduced GC content by 47 and 49% for LaccGluc-Stop and LaccPost-Stop genes, respectively. A codon adaptation index of 0.84 was obtained for both genes. 3D structure analysis using SuperPose revealed LaccGluc-Stop is similar to the laccase crystallographic structure 1GYC of Trametes versicolor. Interaction analysis of the 3D models validated through ABTS, demonstrated higher substrate affinity for LaccPost-Stop, in agreement with our experimental results with enzymatic activities of 451.08 ± 6.46 UL-1 compared to activities of 0.13 ± 0.028 UL-1 for LaccGluc-Stop. This study demonstrated that G. lucidum GlLCC1 and P. ostreatus POXA 1B gene optimization resulted in constitutive gene expression under GAP promoter and α-factor leader in P. pastoris. These are important findings in light of recombinant enzyme expression system utility for environmentally friendly designed expression systems, because of the wide range of substrates

  2. Computational analysis and low-scale constitutive expression of laccases synthetic genes GlLCC1 from Ganoderma lucidum and POXA 1B from Pleurotus ostreatus in Pichia pastoris.

    Science.gov (United States)

    Rivera-Hoyos, Claudia M; Morales-Álvarez, Edwin David; Poveda-Cuevas, Sergio Alejandro; Reyes-Guzmán, Edwin Alfredo; Poutou-Piñales, Raúl A; Reyes-Montaño, Edgar Antonio; Pedroza-Rodríguez, Aura Marina; Rodríguez-Vázquez, Refugio; Cardozo-Bernal, Ángela M

    2015-01-01

    Lacasses are multicopper oxidases that can catalyze aromatic and non-aromatic compounds concomitantly with reduction of molecular oxygen to water. Fungal laccases have generated a growing interest due to their biotechnological potential applications, such as lignocellulosic material delignification, biopulping and biobleaching, wastewater treatment, and transformation of toxic organic pollutants. In this work we selected fungal genes encoding for laccase enzymes GlLCC1 in Ganoderma lucidum and POXA 1B in Pleurotus ostreatus. These genes were optimized for codon use, GC content, and regions generating secondary structures. Laccase proposed computational models, and their interaction with ABTS [2, 2'-azino-bis(3-ethylbenzothiazoline-6-sulphonic acid)] substrate was evaluated by molecular docking. Synthetic genes were cloned under the control of Pichia pastoris glyceraldehyde-3-phosphate dehydrogenase (GAP) constitutive promoter. P. pastoris X-33 was transformed with pGAPZαA-LaccGluc-Stop and pGAPZαA-LaccPost-Stop constructs. Optimization reduced GC content by 47 and 49% for LaccGluc-Stop and LaccPost-Stop genes, respectively. A codon adaptation index of 0.84 was obtained for both genes. 3D structure analysis using SuperPose revealed LaccGluc-Stop is similar to the laccase crystallographic structure 1GYC of Trametes versicolor. Interaction analysis of the 3D models validated through ABTS, demonstrated higher substrate affinity for LaccPost-Stop, in agreement with our experimental results with enzymatic activities of 451.08 ± 6.46 UL-1 compared to activities of 0.13 ± 0.028 UL-1 for LaccGluc-Stop. This study demonstrated that G. lucidum GlLCC1 and P. ostreatus POXA 1B gene optimization resulted in constitutive gene expression under GAP promoter and α-factor leader in P. pastoris. These are important findings in light of recombinant enzyme expression system utility for environmentally friendly designed expression systems, because of the wide range of substrates

  3. Computational analysis and low-scale constitutive expression of laccases synthetic genes GlLCC1 from Ganoderma lucidum and POXA 1B from Pleurotus ostreatus in Pichia pastoris.

    Directory of Open Access Journals (Sweden)

    Claudia M Rivera-Hoyos

    Full Text Available Lacasses are multicopper oxidases that can catalyze aromatic and non-aromatic compounds concomitantly with reduction of molecular oxygen to water. Fungal laccases have generated a growing interest due to their biotechnological potential applications, such as lignocellulosic material delignification, biopulping and biobleaching, wastewater treatment, and transformation of toxic organic pollutants. In this work we selected fungal genes encoding for laccase enzymes GlLCC1 in Ganoderma lucidum and POXA 1B in Pleurotus ostreatus. These genes were optimized for codon use, GC content, and regions generating secondary structures. Laccase proposed computational models, and their interaction with ABTS [2, 2'-azino-bis(3-ethylbenzothiazoline-6-sulphonic acid] substrate was evaluated by molecular docking. Synthetic genes were cloned under the control of Pichia pastoris glyceraldehyde-3-phosphate dehydrogenase (GAP constitutive promoter. P. pastoris X-33 was transformed with pGAPZαA-LaccGluc-Stop and pGAPZαA-LaccPost-Stop constructs. Optimization reduced GC content by 47 and 49% for LaccGluc-Stop and LaccPost-Stop genes, respectively. A codon adaptation index of 0.84 was obtained for both genes. 3D structure analysis using SuperPose revealed LaccGluc-Stop is similar to the laccase crystallographic structure 1GYC of Trametes versicolor. Interaction analysis of the 3D models validated through ABTS, demonstrated higher substrate affinity for LaccPost-Stop, in agreement with our experimental results with enzymatic activities of 451.08 ± 6.46 UL-1 compared to activities of 0.13 ± 0.028 UL-1 for LaccGluc-Stop. This study demonstrated that G. lucidum GlLCC1 and P. ostreatus POXA 1B gene optimization resulted in constitutive gene expression under GAP promoter and α-factor leader in P. pastoris. These are important findings in light of recombinant enzyme expression system utility for environmentally friendly designed expression systems, because of the wide range

  4. Identification of genes preferentially expressed by highly virulent piscine Streptococcus agalactiae upon interaction with macrophages.

    Directory of Open Access Journals (Sweden)

    Chang-Ming Guo

    Full Text Available Streptococcus agalactiae, long recognized as a mammalian pathogen, is an emerging concern with regard to fish. In this study, we used a mouse model and in vitro cell infection to evaluate the pathogenetic characteristics of S. agalactiae GD201008-001, isolated from tilapia in China. This bacterium was found to be highly virulent and capable of inducing brain damage by migrating into the brain by crossing the blood-brain barrier (BBB. The phagocytosis assays indicated that this bacterium could be internalized by murine macrophages and survive intracellularly for more than 24 h, inducing injury to macrophages. Further, selective capture of transcribed sequences (SCOTS was used to investigate microbial gene expression associated with intracellular survival. This positive cDNA selection technique identified 60 distinct genes that could be characterized into 6 functional categories. More than 50% of the differentially expressed genes were involved in metabolic adaptation. Some genes have previously been described as associated with virulence in other bacteria, and four showed no significant similarities to any other previously described genes. This study constitutes the first step in further gene expression analyses that will lead to a better understanding of the molecular mechanisms used by S. agalactiae to survive in macrophages and to cross the BBB.

  5. Constitution, 30 September 1987.

    Science.gov (United States)

    1987-01-01

    This document reprints provisions of Suriname's 1987 Constitution relating to freedom of movement, equality of the sexes, the right to life, the right to physical integrity, equal opportunity in employment, the family, children, maternity benefits, the right to health care, parental responsibilities, free and compulsory education, illiteracy, and housing. All citizens enjoy freedom of movement within the bounds of the law. All people within the territory may claim protection of their person and property, and discrimination is forbidden on the basis of birth, sex, race, language, religion, education, political beliefs, economic position, or other status. Torture or inhuman treatment and punishment is banned, and the right to life is protected by the law. The state guarantees the right to work, and all employees have the right to equal remuneration for equal work, safe working conditions, and sufficient rest and recreation. The family is protected, and husbands and wives are equal before the law. Children have the right to protection, and working women are entitled to paid maternity leave. The state promotes the right to good health by systematic improvements in living and working conditions and dissemination of health education. The right to education is protected by the provision of free general primary education and efforts of the state to enable all citizens to achieve the highest educational levels possible. The Constitution also calls for the institution of a plan to allow the state to create public housing.

  6. Sex-Specific Effects of High Yolk Androgen Levels on Constitutive and Cell-Mediated Immune Responses in Nestlings of an Altricial Passerine

    OpenAIRE

    Muriel, Jaime; Pérez-Rodríguez, Lorenzo; Ortíz-Santaliestra, Manuel E.; Puerta, Marisa; Gil, Diego

    2017-01-01

    Avian embryos are exposed to yolk androgens that are incorporated into the egg by the ovulating female. These steroids can affect several aspects of embryo development, often resulting in increases in overall size or the speed of growth of different traits. However, several studies suggest that they also entail immune costs to the offspring. In this study, we explored whether variation in yolk androgen concentration affected several measures of the constitutive and cell-m...

  7. Multigroup Equivalence Analysis for High-Dimensional Expression Data

    Science.gov (United States)

    Yang, Celeste; Bartolucci, Alfred A.; Cui, Xiangqin

    2015-01-01

    Hypothesis tests of equivalence are typically known for their application in bioequivalence studies and acceptance sampling. Their application to gene expression data, in particular high-dimensional gene expression data, has only recently been studied. In this paper, we examine how two multigroup equivalence tests, the F-test and the range test, perform when applied to microarray expression data. We adapted these tests to a well-known equivalence criterion, the difference ratio. Our simulation results showed that both tests can achieve moderate power while controlling the type I error at nominal level for typical expression microarray studies with the benefit of easy-to-interpret equivalence limits. For the range of parameters simulated in this paper, the F-test is more powerful than the range test. However, for comparing three groups, their powers are similar. Finally, the two multigroup tests were applied to a prostate cancer microarray dataset to identify genes whose expression follows a prespecified trajectory across five prostate cancer stages. PMID:26628859

  8. Multigroup Equivalence Analysis for High-Dimensional Expression Data.

    Science.gov (United States)

    Yang, Celeste; Bartolucci, Alfred A; Cui, Xiangqin

    2015-01-01

    Hypothesis tests of equivalence are typically known for their application in bioequivalence studies and acceptance sampling. Their application to gene expression data, in particular high-dimensional gene expression data, has only recently been studied. In this paper, we examine how two multigroup equivalence tests, the F-test and the range test, perform when applied to microarray expression data. We adapted these tests to a well-known equivalence criterion, the difference ratio. Our simulation results showed that both tests can achieve moderate power while controlling the type I error at nominal level for typical expression microarray studies with the benefit of easy-to-interpret equivalence limits. For the range of parameters simulated in this paper, the F-test is more powerful than the range test. However, for comparing three groups, their powers are similar. Finally, the two multigroup tests were applied to a prostate cancer microarray dataset to identify genes whose expression follows a prespecified trajectory across five prostate cancer stages.

  9. MMSET is highly expressed and associated with aggressiveness in neuroblastoma

    DEFF Research Database (Denmark)

    Hudlebusch, Heidi Rye; Skotte, Julie; Santoni-Rugiu, Eric;

    2011-01-01

    tumor types as well. We have performed immunohistochemical staining of tissue microarrays and found that MMSET protein is frequently and highly expressed in neuroblastoma (MMSET positive in 75% of neuroblastomas, n=164). The expression level of MMSET in neuroblastomas was significantly associated...... with poor survival, negative prognostic factors, and metastatic disease. Moreover, a subset of neuroblastomas for which pre- and post-chemotherapy biopsies were available displayed a strong decrease in MMSET protein levels after chemotherapy. In agreement with neuroblastomas becoming more differentiated...... after treatment, we show that retinoic acid-induced differentiation of human neuroblastoma cells in vitro also leads to a strong decrease in MMSET levels. Furthermore, we demonstrate that the high levels of MMSET in normal neural progenitor cells are strongly downregulated during differentiation...

  10. 基于短乳杆菌S层启动子组成型乳酸杆菌表达系统的构建%Construction of constitutive expression system based on Lactobacillus brevis S layer promoter

    Institute of Scientific and Technical Information of China (English)

    李一经; 秦思; 唐丽杰; 葛俊伟; 乔薪瑗; 崔文; 姜艳平

    2013-01-01

    pPG612 is a shuttle vector suitiable for cloning in both E.coli and Lactobacillus.In the current study,we replaced the promoter of pPG612 vector with S-layer protein's promoter SlpA of Lactobaccillus brevis,and thus constructed a type of expression vector of lactic acid bacteria,of which the expressing foreign protein ability was detected with the TGEV N gene as a target gene in three lactic acid bacillus.We further confirmed that the vector was able to express TGEV N gene N with SDS-PAGE and westem blot.In conclusion,SIpA was a constitutive promoter which could be applied to more studies in the near future.%以大肠杆菌和乳酸菌穿梭分泌表达型载体pPG612为基本模型,用短乳杆菌S层启动子SlpA替换载体上gusA启动子,并以TGEVN蛋白基因为目基因构建一种乳酸杆菌组成型表达载体,用以检测SIpA启动子在三种不同乳酸杆菌中表达能力.结果表明,该组成型启动子具有表达外源功能蛋白功能,且具有严格宿主选择性,为进一步利用该载体及组成型启动子进行其他应用奠定了基础.

  11. PD-L1 is not constitutively expressed on Tasmanian devil facial tumor cells but is strongly upregulated in response to IFN-γ and can be expressed in the tumor microenvironment

    Directory of Open Access Journals (Sweden)

    Andrew Steven Flies

    2016-12-01

    Full Text Available The devil facial tumor disease (DFTD is caused by clonal transmissible cancers that have led to a catastrophic decline in the wild Tasmanian devil (Sarcophilus harrisii population. The first transmissible tumor, now termed DFT1, was first discovered in 1996 and has been continually transmitted to new hosts for at least 20 years. In 2015 a second transmissible cancer (DFT2 was discovered in wild devils, and the DFT2 is genetically distinct and independent from the DFT1. Despite the estimated 136,559 base pair substitutions and 14,647 insertions/deletions in the DFT1 genome as compared to two normal devil reference genomes, the allograft tumors are not rejected by the host immune system. Additionally, genome sequencing of two sub-strains of DFT1 detected greater than 15,000 single-base substitutions that were found in only one of the DFT1 sub-strains, demonstrating the transmissible tumors are evolving and that generation of neoantigens is likely ongoing. Recent evidence in human clinical trials suggests that blocking PD-1:PD-L1 interactions promote anti-tumor immune responses and is most effective in cancers with a high number of mutations. We hypothesized that DFTD cells could exploit the PD-1:PD-L1 inhibitory pathway to evade anti-tumor immune responses. We developed recombinant proteins and monoclonal antibodies to provide the first demonstration that PD-1 binds to both PD-L1 and PD-L2 in a non-placental mammal and show that PD-L1 is upregulated in DFTD cells in response to IFN-γ. Immunohistochemistry showed that PD-L1 is rarely expressed in primary tumor masses, but low numbers of PD-L1+ non-tumor cells were detected in the microenvironment of several metastatic tumors. Importantly, in vitro testing suggests that PD-1 binding to PD-L1 and PD-L2 can be blocked by monoclonal antibodies, which could be critical to understanding how the DFT allografts evade the immune system.

  12. High Erk-1 activation and Gadd45a expression as prognostic markers in high risk pediatric haemolymphoproliferative diseases

    Directory of Open Access Journals (Sweden)

    Rondelli Roberto

    2009-03-01

    Full Text Available Abstract Studies on activated cell-signaling pathways responsible for neoplastic transformation are numerous in solid tumors and in adult leukemias. Despite of positive results in the evolution of pediatric hematopoietic neoplasias, there are some high-risk subtypes at worse prognosis. The aim of this study was to asses the expression and activation status of crucial proteins involved in cell-signaling pathways in order to identify molecular alterations responsible for the proliferation and/or escape from apoptosis of leukemic blasts. The quantitative and qualitative expression and activation of Erk-1, c-Jun, Caspase8, and Gadd45a was analyzed, by immunocytochemical (ICC and western blotting methods, in bone marrow blasts of 72 patients affected by acute myeloid leukemia (AML, T-cell acute lymphoblastic leukemia (ALL and stage IV non-Hodgkin Lymphoma (NHL. We found an upregulation of Erk-1, Caspase8, c-Jun, and Gadd45a proteins with a constitutive activation in 95.8%, 91.7%, 86.2%, 83.4% of analyzed specimens, respectively. It is worth noting that all AML patients showed an upregulation of all proteins studied and the high expression of GADD45a was associated to the lowest DFS median (p = 0.04. On univariate analysis, only Erk-1 phosphorylation status was found to be correlated with a significantly shorter 5-years DFS in all disease subgroups (p = 0.033 and the lowest DFS median in ALL/NHL subgroup (p = 0.04. Moreover, the simultaneous activation of multiple kinases, as we found for c-Jun and Erk-1 (r = 0.26; p = 0.025, might synergistically enhance survival and proliferation potential of leukemic cells. These results demonstrate an involvement of these proteins in survival of blast cells and, consequently, on relapse percentages of the different subgroups of patients.

  13. Constitution, 5 May 1989.

    Science.gov (United States)

    1989-01-01

    This document contains provisions of Cambodia's Constitution of May 5, 1989. Article 7 gives men and women equal rights in marriage and the family, calls for monogamous marriages, and affords social protection to mothers and children. Article 8 guides parent-child relationships. The 14th article defines state property, and the 15th gives citizens full rights to own, use, and inherit land. The use of agricultural and forested land can only be changed with permission. Article 22 assigns educational responsibilities to the state, including free elementary education and a gradual expansion of higher education. Adult literacy classes are also promoted. Article 26 guarantees free medical consultations, and article 27 gives women a 90-day paid maternity leave. Breast-feeding women are also given special privileges. Article 33 guarantees the right to pay equity and to social security benefits. Article 36 grants the freedom to travel, the inviolability of homes, and privacy in correspondence of all types.

  14. High levan accumulation in transgenic tobacco plants expressing the Gluconacetobacter diazotrophicus levansucrase gene.

    Science.gov (United States)

    Banguela, Alexander; Arrieta, Juan G; Rodríguez, Raisa; Trujillo, Luis E; Menéndez, Carmen; Hernández, Lázaro

    2011-06-10

    Bacterial levansucrase (EC 2.4.1.10) converts sucrose into non-linear levan consisting of long β(2,6)-linked fructosyl chains with β(2,1) branches. Bacterial levan has wide food and non-food applications, but its production in industrial reactors is costly and low yielding. Here, we report the constitutive expression of Gluconacetobacter diazotrophicus levansucrase (LsdA) fused to the vacuolar targeting pre-pro-peptide of onion sucrose:sucrose 1-fructosyltransferase (1-SST) in tobacco, a crop that does not naturally produce fructans. In the transgenic plants, levan with degree of polymerization above 10(4) fructosyl units was detected in leaves, stem, root, and flowers, but not in seeds. High levan accumulation in leaves led to gradual phenotypic alterations that increased with plant age through the flowering stage. In the transgenic lines, the fructan content in mature leaves varied from 10 to 70% of total dry weight. No oligofructans were stored in the plant organs, although the in vitro reaction of transgenic LsdA with sucrose yielded β(2,1)-linked FOS and levan. Transgenic lines with levan representing up to 30mgg(-1) of fresh leaf weight produced viable seeds and the polymer accumulation remained stable in the tested T1 and T2 progenies. The lsdA-expressing tobacco represents an alternative source of highly polymerized levan.

  15. Constitution, 5 October 1988.

    Science.gov (United States)

    1989-01-01

    This document contains major provisions of the constitution adopted by Brazil on 5 October 1988. This constitution seeks to promote the welfare of all citizens without discrimination. The equality of all citizens is guaranteed, and the equal rights of women are specifically mentioned. Property rights are also guaranteed and defined. Female inmates are granted the right to remain with their children while breast feeding. Workers are guaranteed a minimum wage, a family allowance for dependents, maternity/paternity leave, specific incentives to protect the labor market for women, retirement benefits, free day care for preschool-age children, pay equity, and equal rights between tenured and sporadically employed workers. Agrarian reform provisions are given, including the authority to expropriate land. Social and economic policies to promote health are called for, and public health services are to be decentralized, to be integrated, and to foster community participation. Pension plan and social assistance provisions are outlined as are duties of the state in regard to education. The amount of money to be dedicated to education is set out, and a national educational plan is called for to achieve such goals as the eradication of illiteracy, the universalization of school attendance, the improvement of instruction, and the provision of vocational training. Specific measures are set out to protect and preserve the environment. Family policy deals with issues of marriage, the definition of a family, divorce, the right to family planning services, and the deterrence of domestic violence. Social protection provisions cover mothers and children, handicapped persons, and protection of minors. Finally, the customs and rights of Indians are protected, with special provisions given to protect land tenure and to protect the rights of Indians in water resource development and prospecting and mining activities.

  16. High SHIP2 Expression Indicates Poor Survival in Colorectal Cancer

    Directory of Open Access Journals (Sweden)

    Ju Yang

    2014-01-01

    Full Text Available SH2-containing inositol 5′-phosphatase 2 (SHIP2, which generally regulates insulin signaling, cytoskeleton remodeling, and receptor endocytosis, has been suggested to play a significant role in tumor development and progression. However, the associations between SHIP2 expression and the clinical features to evaluate its clinicopathologic significance in colorectal cancer (CRC have not been determined yet. In the present study, one-step quantitative real-time polymerase chain reaction (qPCR test and immunohistochemistry (IHC analysis with CRC tissue microarrays (TMA were employed to evaluate the mRNA and protein expression of SHIP2 in CRC. The results showed that SHIP2 expression in the mRNA and protein levels was significantly higher in CRC tissues than that in corresponding noncancerous tissues (both P<0.05. The expression of SHIP2 protein in CRC was related to lymph node metastasis (P=0.036, distant metastasis (P=0.001, and overall survival (P=0.009. Kaplan-Meier method and Cox multifactor analysis suggested that high SHIP2 protein level (P=0.040 and positive distant metastasis (P=0.048 were critically associated with the unfavorable survival of CRC patients. The findings suggested that SHIP2 may be identified as a useful prognostic marker in CRC and targeting CRC may provide novel strategy for CRC treatment.

  17. The AFT1 transcriptional factor is differentially required for expression of high-affinity iron uptake genes in Saccharomyces cerevisiae.

    Science.gov (United States)

    Casas, C; Aldea, M; Espinet, C; Gallego, C; Gil, R; Herrero, E

    1997-06-15

    High-affinity iron uptake in Saccharomyces cerevisiae involves the extracytoplasmic reduction of ferric ions by FRE1 and FRE2 reductases. Ferrous ions are then transported across the plasma membrane through the FET3 oxidase-FTR1 permease complex. Expression of the high-affinity iron uptake genes is induced upon iron deprivation. We demonstrate that AFT1 is differentially involved in such regulation. Aft1 protein is required for maintaining detectable non-induced level of FET3 expression and for induction of FRE2 in iron starvation conditions. On the contrary, FRE1 mRNA induction is normal in the absence of Aft1, although the existence of AFT1 point mutations causing constitutive expression of FRE1 (Yamaguchi-Iwai et al., EMBO J. 14: 1231-1239, 1995) indicates that Aft1 may also participate in FRE1 expression in a dispensable way. The alterations in the basal levels of expression of the high-affinity iron uptake genes may explain why the AFT1 mutant is unable to grow on respirable carbon sources. Overexpression of AFT1 leads to growth arrest of the G1 stage of the cell cycle. Aft1 is a transcriptional activator that would be part of the different transcriptional complexes interacting with the promoter of the high-affinity iron uptake genes. Aft1 displays phosphorylation modifications depending on the growth stage of the cells, and it might link induction of genes for iron uptake to other metabolically dominant requirement for cell growth.

  18. Constitutive models in LAME.

    Energy Technology Data Exchange (ETDEWEB)

    Hammerand, Daniel Carl; Scherzinger, William Mark

    2007-09-01

    The Library of Advanced Materials for Engineering (LAME) provides a common repository for constitutive models that can be used in computational solid mechanics codes. A number of models including both hypoelastic (rate) and hyperelastic (total strain) constitutive forms have been implemented in LAME. The structure and testing of LAME is described in Scherzinger and Hammerand ([3] and [4]). The purpose of the present report is to describe the material models which have already been implemented into LAME. The descriptions are designed to give useful information to both analysts and code developers. Thus far, 33 non-ITAR/non-CRADA protected material models have been incorporated. These include everything from the simple isotropic linear elastic models to a number of elastic-plastic models for metals to models for honeycomb, foams, potting epoxies and rubber. A complete description of each model is outside the scope of the current report. Rather, the aim here is to delineate the properties, state variables, functions, and methods for each model. However, a brief description of some of the constitutive details is provided for a number of the material models. Where appropriate, the SAND reports available for each model have been cited. Many models have state variable aliases for some or all of their state variables. These alias names can be used for outputting desired quantities. The state variable aliases available for results output have been listed in this report. However, not all models use these aliases. For those models, no state variable names are listed. Nevertheless, the number of state variables employed by each model is always given. Currently, there are four possible functions for a material model. This report lists which of these four methods are employed in each material model. As far as analysts are concerned, this information is included only for the awareness purposes. The analyst can take confidence in the fact that model has been properly implemented

  19. Legal theology in imposed constitutionalism

    DEFF Research Database (Denmark)

    Abat Ninet, Antoni

    2017-01-01

    The focus of this paper is the question of legitimacy, and how can we consider legitimate an imposed constitution and the subsequent constitutional principles, practices and values that go hand-in-hand with the legal and political acculturation. Constitutional texts around the world are good...

  20. The diffusion of constitutional rights

    NARCIS (Netherlands)

    Goderis, B.V.G.; Versteeg, M.

    2014-01-01

    Constitutions are commonly regarded as uniquely national products, shaped by domestic ideals and politics. This paper develops and empirically investigates a novel hypothesis, which is that constitutions are also shaped by transnational influence, or “diffusion.” Constitutional rights can diffuse th

  1. Twister ribozymes as highly versatile expression platforms for artificial riboswitches

    Science.gov (United States)

    Felletti, Michele; Stifel, Julia; Wurmthaler, Lena A.; Geiger, Sophie; Hartig, Jörg S.

    2016-01-01

    The utilization of ribozyme-based synthetic switches in biotechnology has many advantages such as an increased robustness due to in cis regulation, small coding space and a high degree of modularity. The report of small endonucleolytic twister ribozymes provides new opportunities for the development of advanced tools for engineering synthetic genetic switches. Here we show that the twister ribozyme is distinguished as an outstandingly flexible expression platform, which in conjugation with three different aptamer domains, enables the construction of many different one- and two-input regulators of gene expression in both bacteria and yeast. Besides important implications in biotechnology and synthetic biology, the observed versatility in artificial genetic control set-ups hints at possible natural roles of this widespread ribozyme class. PMID:27670347

  2. Expressing Constitutively Active Rheb in Adult Dorsal Root Ganglion Neurons Enhances the Integration of Sensory Axons that Regenerate Across a Chondroitinase-Treated Dorsal Root Entry Zone Following Dorsal Root Crush.

    Science.gov (United States)

    Wu, Di; Klaw, Michelle C; Kholodilov, Nikolai; Burke, Robert E; Detloff, Megan R; Côté, Marie-Pascale; Tom, Veronica J

    2016-01-01

    While the peripheral branch of dorsal root ganglion neurons (DRG) can successfully regenerate after injury, lesioned central branch axons fail to regrow across the dorsal root entry zone (DREZ), the interface between the dorsal root and the spinal cord. This lack of regeneration is due to the limited regenerative capacity of adult sensory axons and the growth-inhibitory environment at the DREZ, which is similar to that found in the glial scar after a central nervous system (CNS) injury. We hypothesized that transduction of adult DRG neurons using adeno-associated virus (AAV) to express a constitutively-active form of the GTPase Rheb (caRheb) will increase their intrinsic growth potential after a dorsal root crush. Additionally, we posited that if we combined that approach with digestion of upregulated chondroitin sulfate proteoglycans (CSPG) at the DREZ with chondroitinase ABC (ChABC), we would promote regeneration of sensory axons across the DREZ into the spinal cord. We first assessed if this strategy promotes neuritic growth in an in vitro model of the glial scar containing CSPG. ChABC allowed for some regeneration across the once potently inhibitory substrate. Combining ChABC treatment with expression of caRheb in DRG significantly improved this growth. We then determined if this combination strategy also enhanced regeneration through the DREZ after dorsal root crush in adult rats in vivo. After unilaterally crushing C4-T1 dorsal roots, we injected AAV5-caRheb or AAV5-GFP into the ipsilateral C5-C8 DRGs. ChABC or PBS was injected into the ipsilateral dorsal horn at C5-C8 to digest CSPG, for a total of four animal groups (caRheb + ChABC, caRheb + PBS, GFP + ChABC, GFP + PBS). Regeneration was rarely observed in PBS-treated animals, whereas short-distance regrowth across the DREZ was observed in ChABC-treated animals. No difference in axon number or length between the ChABC groups was observed, which may be related to intraganglionic inflammation induced by the

  3. A Highly Expressed High-Molecular-Weight S-Layer Complex of Pelosinus sp. Strain UFO1 Binds Uranium.

    Science.gov (United States)

    Thorgersen, Michael P; Lancaster, W Andrew; Rajeev, Lara; Ge, Xiaoxuan; Vaccaro, Brian J; Poole, Farris L; Arkin, Adam P; Mukhopadhyay, Aindrila; Adams, Michael W W

    2017-02-15

    Cell suspensions of Pelosinus sp. strain UFO1 were previously shown, using spectroscopic analysis, to sequester uranium as U(IV) complexed with carboxyl and phosphoryl group ligands on proteins. The goal of our present study was to characterize the proteins involved in uranium binding. Virtually all of the uranium in UFO1 cells was associated with a heterodimeric protein, which was termed the uranium-binding complex (UBC). The UBC was composed of two S-layer domain proteins encoded by UFO1_4202 and UFO1_4203. Samples of UBC purified from the membrane fraction contained 3.3 U atoms/heterodimer, but significant amounts of phosphate were not detected. The UBC had an estimated molecular mass by gel filtration chromatography of 15 MDa, and it was proposed to contain 150 heterodimers (UFO1_4203 and UFO1_4202) and about 500 uranium atoms. The UBC was also the dominant extracellular protein, but when purified from the growth medium, it contained only 0.3 U atoms/heterodimer. The two genes encoding the UBC were among the most highly expressed genes within the UFO1 genome, and their expressions were unchanged by the presence or absence of uranium. Therefore, the UBC appears to be constitutively expressed and is the first line of defense against uranium, including by secretion into the extracellular medium. Although S-layer proteins were previously shown to bind U(VI), here we showed that U(IV) binds to S-layer proteins, we identified the proteins involved, and we quantitated the amount of uranium bound.

  4. Ontoteleological Constitution of Entrepreneurship

    Directory of Open Access Journals (Sweden)

    Diego Luiz Teixeira Boava

    2017-06-01

    Full Text Available Entrepreneurship is a pluri-disciplinary phenomenon, object of research in several areas of knowledge. However, studies on this theme present approaches that start to consider entrepreneurship as a field of private knowledge in the phase of epistemological construction. In this context, the aim of this investigation is to contribute to the discussions on the theme, through studies on the ontoteleological constitution of entrepreneurship, in propaedeutic character, deflagrating new approaches. Thus, there is a presentation concerning the study of entrepreneurship, which may emphasize its ontical and ontological aspects. In addition, the reason why it is complex to define entrepreneurship is investigated. Subjects regarding the philosophy of entrepreneurship are introduced, seeking to present the bases for an ontoteleological approach to the phenomenon. Such an approach assumes that the finality of the entrepreneurial act relates to the main principles and transformations required into the organization. Finally, it is concluded that man is an entrepreneurial being, the meta-entrepreneur, and his entrepreneurial actions are not determined by external factors, but rather by the condition of his potentiality.

  5. Governmental control of public expenditure in the constitutional State: thoughts regarding General Comptroller sanctioning powers approval on functional administrative responsibility

    Directory of Open Access Journals (Sweden)

    Erika García Cobián Castro

    2013-12-01

    Full Text Available Constitutional doctrine has little addressed on the General Comptroller of the Republic and its controlling function of public resources execution and management. Efforts have been made to explain this situation in other realities, regarding high budgetary, accounting and auditing content needed to carry out auditing activities constitutionally assigned to this kind of entities. Nevertheless, identifying competences of the General Comptroller of the Republic and its nature of autonomous constitutional body shapes an important expression of Constitutional State and the principle of powers division. This entity controls State’s Public Budget execution contributing to constitutional property protection, legality of budget implementation and the appropriate management of public resources, also the «efficiency» of social needs, proper functioning of public administration and prevention of corruption, among others duties. This article analyzes the constitutional mission of the General Comptroller in a Constitutional State promoting the process of given a constitutional status to the legal system in its area of influence, also optimizing constitutional properties protected. For such purpose, the 29622 recent law («Law that modifies the 27785 Organic Law of National Control System and the General Comptroller of the Republic extending powers in order to sanction in accordance of functional administrative responsibility» shall be used as object of constitutional analysis.

  6. CONSTITUTIVE THEORY AND EXPERIMENT ANALYSIS OF HOT FORMING FOR HIGH STRENGTH STEEL%高强度钢板热成形本构理论与实验分析

    Institute of Scientific and Technical Information of China (English)

    马宁; 胡平; 武文华; 申国哲; 郭威

    2011-01-01

    analyze the effective thermo-mechanical parameters and mechanical properties of multi phases during the hot forming. The components of strain and their evolved mechanism for hot forming are investigated. The phase-transformation volume stress and phase-transformation plastic stress are defined and expressed to explain the mechanism of thermal-mechanical-transformation coupled relations. The modified Norton-Hoff flow stress is applied to describe the flow properties of boron steel at high temperatures and the material parameters is obtained by analyzing the measured data of high-temperature tensile tests. Based on the above research,the thermal-mechanical-transformation coupled models are introduced into the constitutive equations of hot forming, and the integrated and incremental constitutive equations are developed, respectively. Numerical simulation of U-shaped hot forming process is implemented and compared with the experimental results. The results prove the validity of the developed constitutive equations.

  7. Constitution, 1989. [Selected provisions].

    Science.gov (United States)

    1989-01-01

    Chapter XII of the Hungarian Constitution, 1989, details the Fundamental Rights and Duties of Citizens. Everyone lawfully within the territory of Hungary has the right to liberty of movement and the freedom to choose his or her residence, except when restricted by law, including the right to leave his or her residence or county. The Republic of Hungary grants asylum to foreign citizens who were persecuted for racial, religious ethnic, linguistic, or political reasons. Men and women shall equally enjoy all civil, political, economic, social and political rights. Mothers are entitled to special care and protection before and after childbirth; women and juveniles are protected at work by special regulations. Every child has the right to special care an assistance from his or her family, the State, and society, for appropriate physical, spiritual, and moral development. Parents shall decide the kind of education their children receive. Hungary grants equal rights to all person within its territories, without regard to race, color, sex, language, religion, political, or other opinion, national, and social origin, property, birth and other status. Prejudicial discrimination shall be severely punished. Everyone has the right to work, to the free choice of employment and profession and to equal pay for equal work. Citizens have the right to social security, including social services necessary in old age, sickness, disability, widowhood, orphanhood an unemployment through no fault of their own. Hungary guarantees the right to culture for its citizens and realized this right by free and compulsory elementary education, by secondary and higher education which is accessible to all on the basis of capacity, and by the financial support of those receiving an education.

  8. Monoamine oxidase A is highly expressed in classical Hodgkin lymphoma.

    Science.gov (United States)

    Li, Pei Chuan; Siddiqi, Imran N; Mottok, Anja; Loo, Eric Y; Wu, Chieh Hsi; Cozen, Wendy; Steidl, Christian; Shih, Jean Chen

    2017-10-01

    Monoamine oxidase A (MAOA) is a mitochondrial enzyme that catalyzes oxidative deamination of neurotransmitters and dietary amines and produces H2 O2 . It facilitates the progression of gliomas and prostate cancer, but its expression and functional relevance have not been studied in lymphoma. Here, we evaluated MAOA in 427 cases of Hodgkin and non-Hodgkin lymphoma and in a spectrum of reactive lymphoid tissues by immunohistochemistry on formalin-fixed, paraffin-embedded specimens. MAOA was expressed by Hodgkin Reed-Sternberg (HRS) cells in the majority of classical Hodgkin lymphomas (cHLs) (181/241; 75%), with 34.8% showing strong expression. Weak MAOA was also noted in a minority of primary mediastinal large B-cell lymphomas (8/47; 17%) and in a mediastinal gray-zone lymphoma. In contrast, no MAOA was found in non-neoplastic lymphoid tissues, nodular lymphocyte-predominant Hodgkin lymphoma (NLPHL; 0/8) or any other non-Hodgkin lymphomas studied (0/123). MAOA was more common in Epstein-Barr virus (EBV)-negative compared to EBV-positive cHL (p Hodgkin-lymphoma-derived cell lines did not. The MAOA inhibitor clorgyline reduced the growth of L1236 cells and U-HO1 cells, and shRNA knockdown of MAOA reduced the growth of L1236 cells. Conversely, ectopic overexpression of MAOA increased the growth of MAOA-negative HDLM2 cells. Combined treatment with clorgyline and ABVD (doxorubicin, bleomycin, vinblastine, dacarbazine) was more effective in reducing cell growth than either regimen alone. In summary, MAOA is highly expressed in cHL and may reflect the distinct biology of this lymphoma. Further studies on the potential utility of MAOA as a diagnostic marker and therapeutic target are warranted. Copyright © 2017 Pathological Society of Great Britain and Ireland. Published by John Wiley & Sons, Ltd. Copyright © 2017 Pathological Society of Great Britain and Ireland. Published by John Wiley & Sons, Ltd.

  9. A nonlinear constitutive model for magnetostrictive materials

    Institute of Scientific and Technical Information of China (English)

    Xin'en Liu; Xiaojing Zheng

    2005-01-01

    A general nonlinear constitutive model is proposed for magnetostrictive materials, based on the important physical fact that a nonlinear part of the elastic strain produced by a pre-stress is related to the magnetic domain rotation or movement and is responsible for the change of the maximum magnetostrictive strain with the pre-stress. To avoid the complicity of determining the tensor function describing the nonlinear elastic strain part, this paper proposes a simplified model by means of linearizing the nonlinear function.For the convenience of engineering applications, the expressions of the 3-D (bulk), 2-D (film) and 1-D (rod) models are, respectively, given for an isotropic material and their applicable ranges are also discussed. By comparison with the experimental data of a Terfenol-D rod, it is found that the proposed model can accurately predict the magnetostrictive strain curves in low, moderate and high magnetic field regions for various compressive pre-stress levels. The numerical simulation further illustrates that, for either magnetostrictive rods or thin films, the proposed model can effectively describe the effects of the pre-stress or residual stress on the magnetization and magnetostrictive strain curves, while none of the known models can capture all of them. Therefore, the proposed model enjoys higher precision and wider applicability than the previous models, especially in the region of the high field.

  10. 小麦籽粒淀粉分支酶同工酶结构组成及时空表达%Constitution and Spatiotemporal Expression of Starch Branching Enzyme in Developing Wheat Grain

    Institute of Scientific and Technical Information of China (English)

    刘正帅; 刘贵芬; 杨明煜; 贾晓; 李运祥; 赵法茂

    2014-01-01

    This study aimed at disclosing the enzymatic mechanism in amylopectin synthesis in wheat (Triticum aestivum L.). The isozyme forms, organ localization, spatiotemporal expression profile and subunits constitution of starch branching enzyme (SBE) were identified in eight wheat cultivars from different provenances using native polyacrylamide gel electrophoresis (Native-PAGE) and SDS-PAGE. Four SBE isozymes were detected in wheat endosperm, in which isozymes B and SBEIIa were localized in en-dosperm and leaf, whereas isozymes A and Di were exclusively present in endosperm. In the process of grain filling, Di and SBEIIa expressed first, followed by isozyme B, and isozyme A expressed finally. However, B and SBEIIa terminated to express at late filling stage. All SBE isozymes were composed of one subunit of 86–92 kD, and their spatial localization exhibited organ specificity. According to the expression level, Di, B, and SBEIIa are considered as dominant isozymes for grain endosperm de-velopment. They probably determinate the total SBE activity and serve as key factors in amylpectin biosynthesis in wheat grain and leaf.%为阐明小麦支链淀粉合成的酶学机制,以8个小麦品种的籽粒为材料,采用非变性聚丙烯酰胺凝胶电泳(Native-PAGE)和 SDS 聚丙烯酰胺凝胶电泳(SDS-PAGE)鉴定 SBE 同工酶类型、时空表达谱及亚基组成,分析 SBE 同工酶空间分布特点和器官表达特异性。共检测到4种 SBE 同工酶,其中 B 和 SBEIIa 分布在胚乳和叶片中,而 A 和Di 专一定位于胚乳中。在小麦籽粒灌浆过程中, Di 和 SBEIIa 首先表达,而后是 B, A 最后表达;至灌浆末期, B 和SBEIIa 停止表达。SBE 同工酶都是单亚基酶,均由一条86~92 kD 的多肽链组成。SBE 同工酶的空间分布具有器官特异性,并在籽粒发育进程中顺序表达。Di、B 和 SBEIIa 是占主导地位的 SBE 同工酶,可能是决定 SBE 总酶活性的主效应酶,在籽粒和叶片支链淀粉合成中起关键作用。

  11. The Political Economy of Constitution

    OpenAIRE

    Pabst, Adrian

    2016-01-01

    The distinction between constitution, as the set of fundamental normative premises ensuring the cohesion of any given polity, and contract, as the formal covenant agreed upon by the relevant stakeholders in that polity, is central to political economy. This paper outlines a conceptual framework for the political economy of constitution based on the above distinction. Our argument is that constitution in the material sense, that is, as a relatively stable configuration of interests prior to fo...

  12. Constitutional Issues--Watergate and the Constitution. Teaching with Documents.

    Science.gov (United States)

    National Archives and Records Administration, Washington, DC.

    When U.S. President Richard Nixon resigned in 1974 in the wake of the Watergate scandal, it was only the second time that impeachment of a president had been considered. Although the U.S. Constitution has provisions for a person removed from office to be indicted, there are no guidelines in the Constitution about a President who has resigned. The…

  13. High-level intracellular expression of heterologous proteins in Brevibacillus choshinensis SP3 under the control of a xylose inducible promoter

    Directory of Open Access Journals (Sweden)

    D’Urzo Nunzia

    2013-02-01

    Full Text Available Abstract Background In past years research has focused on the development of alternative Gram positive bacterial expression systems to produce industrially relevant proteins. Brevibacillus choshinensis is an easy to handle non-sporulating bacterium, lacking extracellular proteases, that has been already shown to provide a high level of recombinant protein expression. One major drawback, limiting the applicability of the Brevibacillus expression system, is the absence of expression vectors based on inducible promoters. Here we used the PxylA inducible promoter, commonly employed in other Bacillae expression systems, in Brevibacillus. Results Using GFP, α-amylase and TcdA-GT as model proteins, high level of intracellular protein expression (up to 250 mg/L for the GFP was achieved in Brevibacillus, using the pHis1522 vector carrying the B. megaterium xylose-inducible promoter (PxylA. The GFP expression yields were more than 25 fold higher than those reported for B. megaterium carrying the same vector. All the tested proteins show significant increment in their expression levels (2-10 folds than those obtained using the available plasmids based on the P2 constitutive promoter. Conclusion Combining the components of two different commercially available Gram positive expression systems, such as Brevibacillus (from Takara Bio and B. megaterium (from Mobitec, we demonstrate that vectors based on the B. megaterium PxylA xylose inducible promoter can be successfully used to induce high level of intracellular expression of heterologous proteins in Brevibacillus.

  14. The Spanish Constitution, the Constitutional Court and the Catalan Referendum

    DEFF Research Database (Denmark)

    Abat Ninet, Antoni

    2015-01-01

    This chapter deals with a core topic in constitutional law, that of the conflict between constitutions and will of the demos. Aristotle in Book IV of the Politics had already anticipated the possible conflict between these two forms of Politeia (constitutions), when defining the types of democracy....... He also distinguished the role that the Nomophylakes (guardians of the laws) should play in this sort of anticipated conflict after what Madison v. Marbury called had globally referred to it around the world as “judicial review”. But the concept of a constitution as a form of government......; and as a document that organised the offices of a state changed through the history. The Magna Carta, the French and American revolutionaries provided gave the concept a new meaning to the concept. After these historical episodes, constitutions were not simply political or legal norms to knowshowing how to play...

  15. Sex-Specific Effects of High Yolk Androgen Levels on Constitutive and Cell-Mediated Immune Responses in Nestlings of an Altricial Passerine.

    Science.gov (United States)

    Muriel, Jaime; Pérez-Rodríguez, Lorenzo; Ortiz-Santaliestra, Manuel E; Puerta, Marisa; Gil, Diego

    Avian embryos are exposed to yolk androgens that are incorporated into the egg by the ovulating female. These steroids can affect several aspects of embryo development, often resulting in increases in overall size or the speed of growth of different traits. However, several studies suggest that they also entail immune costs to the offspring. In this study, we explored whether variation in yolk androgen concentration affected several measures of the constitutive and cell-mediated immune axes in the spotless starling (Sturnus unicolor). Using a within-brood design, we injected different doses of androgens (testosterone and androstenedione) into the eggs. Our study showed that experimentally increased yolk androgens led to sex-specific immunosuppression in both the innate and adaptive axes of the immune system. Both cell-mediated immune response (CMI) and lysozyme activity decreased with increasing androgen levels injected into the egg in the case of male nestlings, whereas there were no effects on females. The effects that we found were always linear: no quadratic or threshold patterns were detected. We found no effects of the experimental treatment in hemolysis or agglutination capacity, but these measures were negatively correlated with CMI, suggesting negative correlation among different branches of the immune system. Blood (trypanosomes and hemosporidians) and intestinal (coccidia) parasites were not affected by the experimental increase of yolk androgen levels. Our results show that in our study species yolk androgens induce immunosuppression in some axes of the male nestling immune system. Further studies should analyze the proximate causes for these contrasting effects in different axes of the immune system and the reason for the differential impact on males and females.

  16. High pressure-sensitive gene expression in Lactobacillus sanfranciscensis

    Directory of Open Access Journals (Sweden)

    R.F. Vogel

    2005-08-01

    Full Text Available Lactobacillus sanfranciscensis is a Gram-positive lactic acid bacterium used in food biotechnology. It is necessary to investigate many aspects of a model organism to elucidate mechanisms of stress response, to facilitate preparation, application and performance in food fermentation, to understand mechanisms of inactivation, and to identify novel tools for high pressure biotechnology. To investigate the mechanisms of the complex bacterial response to high pressure we have analyzed changes in the proteome and transcriptome by 2-D electrophoresis, and by microarrays and real time PCR, respectively. More than 16 proteins were found to be differentially expressed upon high pressure stress and were compared to those sensitive to other stresses. Except for one apparently high pressure-specific stress protein, no pressure-specific stress proteins were found, and the proteome response to pressure was found to differ from that induced by other stresses. Selected pressure-sensitive proteins were partially sequenced and their genes were identified by reverse genetics. In a transcriptome analysis of a redundancy cleared shot gun library, about 7% of the genes investigated were found to be affected. Most of them appeared to be up-regulated 2- to 4-fold and these results were confirmed by real time PCR. Gene induction was shown for some genes up-regulated at the proteome level (clpL/groEL/rbsK, while the response of others to high hydrostatic pressure at the transcriptome level seemed to differ from that observed at the proteome level. The up-regulation of selected genes supports the view that the cell tries to compensate for pressure-induced impairment of translation and membrane transport.

  17. A Highly Expressed High-Molecular-Weight S-Layer Complex of Pelosinus sp. Strain UFO1 Binds Uranium

    Energy Technology Data Exchange (ETDEWEB)

    Thorgersen, Michael P. [Univ. of Georgia, Athens, GA (United States). Dept. of Biochemistry and Molecular Biology; Lancaster, W. Andrew [Univ. of Georgia, Athens, GA (United States). Dept. of Biochemistry and Molecular Biology; Rajeev, Lara [Lawrence Berkeley National Lab. (LBNL), Berkeley, CA (United States). Biological Systems and Engineering Division; Ge, Xiaoxuan [Univ. of Georgia, Athens, GA (United States). Dept. of Biochemistry and Molecular Biology; Vaccaro, Brian J. [Univ. of Georgia, Athens, GA (United States). Dept. of Biochemistry and Molecular Biology; Poole, Farris L. [Univ. of Georgia, Athens, GA (United States). Dept. of Biochemistry and Molecular Biology; Arkin, Adam P. [Lawrence Berkeley National Lab. (LBNL), Berkeley, CA (United States). Biological Systems and Engineering Division; Mukhopadhyay, Aindrila [Lawrence Berkeley National Lab. (LBNL), Berkeley, CA (United States). Biological Systems and Engineering Division; Adams, Michael W. W. [Univ. of Georgia, Athens, GA (United States). Dept. of Biochemistry and Molecular Biology

    2016-12-02

    Cell suspensions of Pelosinus sp. strain UFO1 were previously shown, using spectroscopic analysis, to sequester uranium as U(IV) complexed with carboxyl and phosphoryl group ligands on proteins. The goal of our present study was to characterize the proteins involved in uranium binding. Virtually all of the uranium in UFO1 cells was associated with a heterodimeric protein, which was termed the uranium-binding complex (UBC). The UBC was composed of two S-layer domain proteins encoded by UFO1_4202 and UFO1_4203. Samples of UBC purified from the membrane fraction contained 3.3 U atoms/heterodimer, but significant amounts of phosphate were not detected. The UBC had an estimated molecular mass by gel filtration chromatography of 15 MDa, and it was proposed to contain 150 heterodimers (UFO1_4203 and UFO1_4202) and about 500 uranium atoms. The UBC was also the dominant extracellular protein, but when purified from the growth medium, it contained only 0.3 U atoms/heterodimer. The two genes encoding the UBC were among the most highly expressed genes within the UFO1 genome, and their expressions were unchanged by the presence or absence of uranium. Therefore, the UBC appears to be constitutively expressed and is the first line of defense against uranium, including by secretion into the extracellular medium. Although S-layer proteins were previously shown to bind U(VI), here we showed that U(IV) binds to S-layer proteins, we identified the proteins involved, and we quantitated the amount of uranium bound. Widespread uranium contamination from industrial sources poses hazards to human health and to the environment. Here in this paper, we identified a highly abundant uranium-binding complex (UBC) from Pelosinus sp. strain UFO1. The complex makes up the primary protein component of the S-layer of strain UFO1 and binds 3.3 atoms of U(IV) per heterodimer. Finally, while other bacteria have been shown to bind U(VI) on their S-layer, we demonstrate here an example of U(IV) bound by

  18. Constitutional Law and Liberal Education.

    Science.gov (United States)

    Clor, Harry

    1985-01-01

    By studying constitutional law, students learn about the relationship between democratic theory and practice, one of the main concerns of liberal education. The mind is enlarged when it must apply ethical standards and political ideas to real human problems. How a political science professor teaches constitutional law is discussed. (RM)

  19. Reordering American Constitutional Law Teaching.

    Science.gov (United States)

    Gerber, Scott D.

    1994-01-01

    Maintains that constitutional law is the cornerstone of an undergraduate public law curriculum. Asserts that there is a welcome trend toward teaching the subject over a two-semester sequence, instead of only one. Describes course content and teaching strategies used in a college constitutional law course. (CFR)

  20. The Constitution and Its Critics

    Science.gov (United States)

    Main, Thomas J.

    2011-01-01

    In planning a freshman undergraduate curriculum with colleagues recently, the question arose as to what type of understanding educators wanted to impart to their students about the Constitution. The alleged defects of the Constitution that these books point to are wide-ranging and can be classified into various categories. Some problems--such as…

  1. Constitutional developments in Latin American abortion law.

    Science.gov (United States)

    Bergallo, Paola; Ramón Michel, Agustina

    2016-11-01

    For most of the 20th Century, restrictive abortion laws were in place in continental Latin America. In recent years, reforms have caused a liberalizing shift, supported by constitutional decisions of the countries' high courts. The present article offers an overview of the turn toward more liberal rules and the resolution of abortion disputes by reference to national constitutions. For such purpose, the main legal changes of abortion laws in the last decade are first surveyed. Landmark decisions of the high courts of Argentina, Bolivia, Colombia, and Mexico are then analyzed. It is shown that courts have accepted the need to balance interests and competing rights to ground less restrictive laws. In doing so, they have articulated limits to protection of fetal interests, and basic ideas of women's dignity, autonomy, and equality. The process of constitutionalization has only just begun. Constitutional judgments are not the last word, but they are important contributions in reinforcing the legality of abortion.

  2. The constitutive sofa cushion

    DEFF Research Database (Denmark)

    Hanghøj, Sara

    2009-01-01

    , with the sitting room as a cultural and informal environment as its arena in 2006. The local and social network of the informant (born 1919) is traced through a detailed cultural analysis of the creation, usage and existence of the sofa cushion, including even the material interactions occurring between the human...... body, space and artefact, thereby revealing the materialization process. Based on results of the analysis, ANT (Actor-Network- Theory) is used in order to discuss how The Danish Folk High School's conception of simplicity as an aesthetic and gendered ideal in the formal education of female handcraft...

  3. Expression

    Directory of Open Access Journals (Sweden)

    Wang-Xia Wang

    2014-02-01

    Full Text Available The miR-15/107 family comprises a group of 10 paralogous microRNAs (miRNAs, sharing a 5′ AGCAGC sequence. These miRNAs have overlapping targets. In order to characterize the expression of miR-15/107 family miRNAs, we employed customized TaqMan Low-Density micro-fluid PCR-array to investigate the expression of miR-15/107 family members, and other selected miRNAs, in 11 human tissues obtained at autopsy including the cerebral cortex, frontal cortex, primary visual cortex, thalamus, heart, lung, liver, kidney, spleen, stomach and skeletal muscle. miR-103, miR-195 and miR-497 were expressed at similar levels across various tissues, whereas miR-107 is enriched in brain samples. We also examined the expression patterns of evolutionarily conserved miR-15/107 miRNAs in three distinct primary rat brain cell preparations (enriched for cortical neurons, astrocytes and microglia, respectively. In primary cultures of rat brain cells, several members of the miR-15/107 family are enriched in neurons compared to other cell types in the central nervous system (CNS. In addition to mature miRNAs, we also examined the expression of precursors (pri-miRNAs. Our data suggested a generally poor correlation between the expression of mature miRNAs and their precursors. In summary, we provide a detailed study of the tissue and cell type-specific expression profile of this highly expressed and phylogenetically conserved family of miRNA genes.

  4. High-level expressing YAC vector for transgenic animal bioreactors.

    Science.gov (United States)

    Fujiwara, Y; Miwa, M; Takahashi, R; Kodaira, K; Hirabayashi, M; Suzuki, T; Ueda, M

    1999-04-01

    The position effect is one major problem in the production of transgenic animals as mammary gland bioreactors. In the present study, we introduced the human growth hormone (hGH) gene into 210-kb human alpha-lactalbumin position-independent YAC vectors using homologous recombination and produced transgenic rats via microinjection of YAC DNA into rat embryos. The efficiency of producing transgenic rats with the YAC vector DNA was the same as that using plasmid constructs. All analyzed transgenic rats had one copy of the transgene and produced milk containing a high level of hGH (0.25-8.9 mg/ml). In transgenic rats with the YAC vector in which the human alpha-lactalbumin gene was replaced with the hGH gene, tissue specificity of hGH mRNA was the same as that of the endogenous rat alpha-lactalbumin gene. Thus, the 210-kb human alpha-lactalbumin YAC is a useful vector for high-level expression of foreign genes in the milk of transgenic animals.

  5. Intensity dependence in high-level facial expression adaptation aftereffect.

    Science.gov (United States)

    Hong, Sang Wook; Yoon, K Lira

    2017-06-14

    Perception of a facial expression can be altered or biased by a prolonged viewing of other facial expressions, known as the facial expression adaptation aftereffect (FEAA). Recent studies using antiexpressions have demonstrated a monotonic relation between the magnitude of the FEAA and adaptor extremity, suggesting that facial expressions are opponent coded and represented continuously from one expression to its antiexpression. However, it is unclear whether the opponent-coding scheme can account for the FEAA between two facial expressions. In the current study, we demonstrated that the magnitude of the FEAA between two facial expressions increased monotonically as a function of the intensity of adapting facial expressions, consistent with the predictions based on the opponent-coding model. Further, the monotonic increase in the FEAA occurred even when the intensity of an adapting face was too weak for its expression to be recognized. These results together suggest that multiple facial expressions are encoded and represented by balanced activity of neural populations tuned to different facial expressions.

  6. Internal constitution of Mars.

    Science.gov (United States)

    Anderson, D. L.

    1972-01-01

    Models of the internal structure of Mars consistent with the mass, radius and moment of inertia of the planet are constructed. The models assume that the radius of the core is between 0.36 and 0.60 of the radius of the planet, that the zero-pressure density of the mantle is between 3.54 and 3.49 g/cu cm, and that the planet contains 25 to 28% iron. Meteorite models of Mars containing 25 wt % iron and 12 wt % core are also proposed. It is maintained that Mars in contrast to the earth is an incompletely differentiated planet with a core substantially richer in sulfur than the core of the earth. The absence of a magnetic field on Mars is possibly linked with lack of lunar precessional torque and the small size and high resistivity of the Martian core.

  7. Report between the President and Constitutional Court and its influence on the functioning of the Constitutional System in Kosovo

    Directory of Open Access Journals (Sweden)

    Florent Muçaj

    2016-07-01

    Full Text Available This paper aims at clarifying the report between the President and the Constitutional Court. If we take as a starting point the constitutional mandate of these two institutions it follows that their final mission is the same, i.e., the protection and safeguarding of the constitutional system. This paper, thus, will clarify the key points in which this report is expressed. Further, this paper examines the theoretical aspects of the report between the President and the Constitutional Court, starting from the debate over this issue between Karl Schmitt and Hans Kelsen. An important part of the paper will examine the Constitution of Kosovo, i.e., the contents of the constitutional norm and its application. The analysis focuses on the role such report between the two institutions has on the functioning of the constitutional system. In analyzing the case of Kosovo, this paper examines Constitutional Court cases in which the report between the President and the Constitutional Court has been an issue of review. Such cases assist us in clarifying the main theme of this paper. Therefore, the reader will be able to understand the key elements of the report between the President as a representative of the unity of the people on the one hand and the Constitutional Court as a guarantor of constitutionality on the other hand.

  8. Rapping the 27 Amendments to the Constitution

    Science.gov (United States)

    Knaresborough, Adam

    2009-01-01

    Early in the year, the students of history and government at Mountain View High School in Stafford, Virginia, began to devise hand motions to help memorize the 27 amendments to the Constitution for government class. Three students in the school who are interested in hip hop music then suggested composing a rap song about the topic. Working with…

  9. High heregulin expression is associated with activated HER3 and may define an actionable biomarker in patients with squamous cell carcinomas of the head and neck.

    Directory of Open Access Journals (Sweden)

    David S Shames

    Full Text Available PURPOSE: Tumors with oncogenic dependencies on the HER family of receptor tyrosine kinases (RTKs often respond well to targeted inhibition. Our previous work suggested that many cell lines derived from squamous cell carcinomas of the head and neck (SCCHNs depend on autocrine signaling driven by HER2/3 dimerization and high-level co-expression of HRG. Additionally, results from a Phase I trial of MEHD7495A, a dual-action antibody that blocks ligand binding to EGFR and HER3, suggest that high-level HRG expression was associated with clinical response in SCCHN patients. Here we explore the hypothesis that high-level HRG expression defines a subpopulation of SCCHNs with activated HER3. EXPERIMENTAL DESIGN: qRT-PCR expression profiling was performed on >750 tumors of diverse origin, including >150 therapy-naïve, primary, and recurrent SCCHNs. Activated HER3, defined by immunoprecipitation of phospho-HER3, was compared to HRG expression in SCCHN samples. Paracrine versus autocrine expression was evaluated using RNA-in situ hybridization. RESULTS: SCCHN tumors express the highest levels of HRG compared to a diverse collection of other tumor types. We show that high HRG expression is associated with activated HER3, whereas low HRG expression is associated with low HER3 activation in SCCHN tumors. Furthermore, HRG expression is higher in recurrent SCCHN compared to patient-matched therapy naïve specimens. CONCLUSIONS: HRG expression levels define a biologically distinct subset of SCCHN patients. We propose that high-level expression of HRG is associated with constitutive activation of HER3 in SCCHN and thus defines an actionable biomarker for interventions targeting HER3.

  10. High constitutive activity of a virus-encoded seven transmembrane receptor in the absence of the conserved DRY motif (Asp-Arg-Tyr) in transmembrane helix 3

    DEFF Research Database (Denmark)

    Rosenkilde, Mette M; Kledal, Thomas N; Schwartz, Thue W

    2005-01-01

    The highly conserved Arg in the so-called DRY motif (Asp-Arg-Tyr) at the intracellular end of transmembrane helix 3 is in general considered as an essential residue for G protein coupling in rhodopsin-like seven transmembrane (7TM) receptors. In the open reading frame 74 (ORF74) receptor encoded ...

  11. Glut4 Is Sorted from a Rab10 GTPase-independent Constitutive Recycling Pathway into a Highly Insulin-responsive Rab10 GTPase-dependent Sequestration Pathway after Adipocyte Differentiation.

    Science.gov (United States)

    Brewer, Paul Duffield; Habtemichael, Estifanos N; Romenskaia, Irina; Mastick, Cynthia Corley; Coster, Adelle C F

    2016-01-08

    The RabGAP AS160/TBC1D4 controls exocytosis of the insulin-sensitive glucose transporter Glut4 in adipocytes. Glut4 is internalized and recycled through a highly regulated secretory pathway in these cells. Glut4 also cycles through a slow constitutive endosomal pathway distinct from the fast transferrin (Tf) receptor recycling pathway. This slow constitutive pathway is the only Glut4 cycling pathway in undifferentiated fibroblasts. The α2-macroglobulin receptor LRP1 cycles with Glut4 and the Tf receptor through all three exocytic pathways. To further characterize these pathways, the effects of knockdown of AS160 substrates on the trafficking kinetics of Glut4, LRP1, and the Tf receptor were measured in adipocytes and fibroblasts. Rab10 knockdown decreased cell surface Glut4 in insulin-stimulated adipocytes by 65%, but not in basal adipocytes or in fibroblasts. This decrease was due primarily to a 62% decrease in the rate constant of Glut4 exocytosis (kex), although Rab10 knockdown also caused a 1.4-fold increase in the rate constant of Glut4 endocytosis (ken). Rab10 knockdown in adipocytes also decreased cell surface LRP1 by 30% by decreasing kex 30-40%. There was no effect on LRP1 trafficking in fibroblasts or on Tf receptor trafficking in either cell type. These data confirm that Rab10 is an AS160 substrate that limits exocytosis through the highly insulin-responsive specialized secretory pathway in adipocytes. They further show that the slow constitutive endosomal (fibroblast) recycling pathway is Rab10-independent. Thus, Rab10 is a marker for the specialized pathway in adipocytes. Interestingly, mathematical modeling shows that Glut4 traffics predominantly through the specialized Rab10-dependent pathway both before and after insulin stimulation.

  12. The significance of constitutional values

    Directory of Open Access Journals (Sweden)

    HN Nisihara

    2001-12-01

    Full Text Available This article addresses the question of the meaning and legal significance of constitutional values in contemporary times. The article attends also to related questions namely, what constituteconstitutional values” and what are the limitations of the meaning afforded to this notion. Attention is paid in the particular, to freedom, equality and democracy as value-neutral criteria of fairness and government neutrality with reference to the South African and German contexts as well as to value-neutrality as a culturally conditioned value. The author concludes with a cosmopolitan view of freedom and the right to peace with reference to the constitutional texts of Japan and the United States.

  13. Are Sanctions on Employers Constitutional?

    Science.gov (United States)

    Gollobin, Ira

    1988-01-01

    Questions the constitutional validity of employer sanctions used to deter illegal immigration under the Immigration Reform and Control Act. Points out the anomaly of using criminal penalties to deter a civil, administrative violation. (FMW)

  14. The President and the Constitution.

    Science.gov (United States)

    Toler, Frank

    1988-01-01

    Intended for ninth grade students, this ten day unit focuses on the constitutional powers of the President of the United States. Included are worksheets, vocabulary, writing assignments, tests, and quizzes. (JDH)

  15. Constitutive Modelling in Geomechanics Introduction

    CERN Document Server

    Puzrin, Alexander M

    2012-01-01

    The purpose of this book is to bridge the gap between the traditional Geomechanics and Numerical Geotechnical Modelling with applications in science and practice. Geomechanics is rarely taught within the rigorous context of Continuum Mechanics and Thermodynamics, while when it comes to Numerical Modelling, commercially available finite elements or finite differences software utilize constitutive relationships within the rigorous framework. As a result, young scientists and engineers have to learn the challenging subject of constitutive modelling from a program manual and often end up with using unrealistic models which violate the Laws of Thermodynamics.  The book is introductory, by no means does it claim any completeness and state of the art in such a dynamically developing field as numerical and constitutive modelling of soils. The author gives basic understanding of conventional continuum mechanics approaches to constitutive modelling, which can serve as a foundation for exploring more advanced theories....

  16. Jonathan's Constitutional Conference in Nigeria

    African Journals Online (AJOL)

    ian

    2013-10-01

    Oct 1, 2013 ... Reasonably and pragmatically too, democratic-driven processes, in which .... Platonic conception of elite's rationality, the theory posits that the domain of ... 'the people' are hardly the players in constitution-making and design.

  17. Are Sanctions on Employers Constitutional?

    Science.gov (United States)

    Gollobin, Ira

    1988-01-01

    Questions the constitutional validity of employer sanctions used to deter illegal immigration under the Immigration Reform and Control Act. Points out the anomaly of using criminal penalties to deter a civil, administrative violation. (FMW)

  18. Isolation and functional characterization of a cotton ubiquitination-related promoter and 5'UTR that drives high levels of expression in root and flower tissues

    Directory of Open Access Journals (Sweden)

    Viana Antonio AB

    2011-11-01

    Full Text Available Abstract Background Cotton (Gossypium spp. is an important crop worldwide that provides raw material to 40% of the textile fiber industry. Important traits have been studied aiming the development of genetically modified crops including resistance to insect and diseases, and tolerance to drought, cold and herbicide. Therefore, the characterization of promoters and regulatory regions is also important to achieve high gene expression and/or a specific expression pattern. Commonly, genes involved in ubiquitination pathways are highly and differentially expressed. In this study, we analyzed the expression of a cotton ubiquitin-conjugating enzyme (E2 family member with no previous characterization. Results Nucleotide analysis revealed high identity with cotton E2 homologues. Multiple alignment showed a premature stop codon, which prevents the encoding of the conserved cysteine residue at the E2 active site, and an intron that is spliced in E2 homologues, but not in GhGDRP85. The GhGDRP85 gene is highly expressed in different organs of cotton plants, and has high transcript levels in roots. Its promoter (uceApro2 and the 5'UTR compose a regulatory region named uceA1.7, and were isolated from cotton and studied in Arabidopsis thaliana. uceA1.7 shows strong expression levels, equaling or surpassing the expression levels of CaMV35S. The uceA1.7 regulatory sequence drives GUS expression 7-fold higher in flowers, 2-fold in roots and at similar levels in leaves and stems. GUS expression levels are decreased 7- to 15-fold when its 5'UTR is absent in uceApro2. Conclusions uceA1.7 is a strong constitutive regulatory sequence composed of a promoter (uceApro2 and its 5'UTR that will be useful in genetic transformation of dicots, having high potential to drive high levels of transgene expression in crops, particularly for traits desirable in flower and root tissues.

  19. CONSTITUTIONALISM

    African Journals Online (AJOL)

    eliasn

    identified as a criminal act by the legislature through the law-making process enshrined in a ... or under the law without discrimination on the basis of status, wealth, race, nationality, gender .... 23 Wikipedia, the free encyclopedia. 24 Id. 25 Id.

  20. STABILITY AND CONSTITUTIONAL REFORM NORMATIVE CONTENTS OF CONSTITUTION

    Directory of Open Access Journals (Sweden)

    Marius ANDREESCU

    2016-05-01

    Full Text Available The modification of the fundamental law of a state represents a very special political and juridical act with major significances and implications in the political social system as in the state’s one, but also at each individual level. That’s why such an approach needs to be well justified, to answer to some juridical and political social needs well defined, but mainly to correspond to the principles and rules specific to a constitutional and state’s democratic system providing to the state the stability and functionality it needs. In this study we analyze the necessity of such a constitutional reform in Romania, and also some provisions from the report of the Presidential Commission for the analysis of the political and constitutional regime in our country. We formulate our opinions in relation to the justifying some constitutional regulations. In this context, we consider that there are arguments for the maintaining of the bicameral parliamentary system and an eventual revising of the fundamental law needs to consider the measures needed to guarantee the political and constitutional institutions specific to the lawful state.

  1. Establishment of Mammalian Cell Lines for Constitutive Expression of Influenza Virus Matrix Protein 2%稳定表达流感病毒基质蛋白2的哺乳动物细胞系的建立

    Institute of Scientific and Technical Information of China (English)

    陈爱珺; 郭建强; 姚立红; 殷继永; 张智清

    2013-01-01

    To establish a mammalian cell line for stable expression of the matrix protein 2 (M2) of influenza virus type A. M2 gene was amplified by PCR from the influenza virus strain A/PR/8/34. The PCR product was cloned into eukaryotic expression vector pcDNA5/FRT. After identification with restriction enzyme digestion, the plasmid was co-transfected with plasmid pOG44 which expressed Flp in Flp-In-CHO cells. The target gene was integrated into chromosome of CHO cells by homologous recombination in vivo. Recombinant CHO-M2 cell lines were selected for hygromycin B resistance. A total of 15 recombinant cell strains with high expression of M2 protein were screened by hygromycin, and the expression of M2 protein was determined by IFA and Western blot. After subculturing for 10 passages, the presence of M2 gene in the CHO-M2 cells was confirmed by PCR, and the expression of M2 protein were proved by IFA and Western blot. We successfully constructed a mammalian cell line which stably expressed M2 protein of influenza virus type A. The cell line will be useful for studies on function of M2 protein and provide tools for novel nfluenza virus vaccine development.%本研究构建了稳定表达甲型流感病毒基质蛋白2(M2)的哺乳动物细胞系.应用PCR方法扩增A/PR/8/34(H1N1)株流感病毒M2基因,将其克隆至真核表达载体pcDNA5/FRT(pDF)上,构建出pDF-M2重组质粒.将鉴定正确的pDF M2与表达Flp重组酶的pOG44质粒共转染Flp-In-CHO细胞,通过体内同源重组使目的基因整合到宿主细胞染色体上.筛选具有Hygromycin B抗性的重组细胞株命名为CHO-M2.以间接免疫荧光法(IFA)和Western blot法检测M2的表达,共获得15株高表达M2蛋白的重组细胞株.这些重组细胞株在连续培养10代后,PCR方法仍可检测到M2基因的存在,IFA也检测到蛋白的稳定表达.本研究成功获得了稳定表达甲型流感病毒M2的哺乳动物细胞系,为M2蛋白的功能研究和非复制型流感病毒疫苗的研制提供了新的工具.

  2. High expression of markers of apoptosis in Langerhans cell histiocytosis

    DEFF Research Database (Denmark)

    Petersen, Bodil Laub; Lundegaard, Pia Rengtved; Bank, M I;

    2003-01-01

    53 and the number of cells in apoptosis detected with TUNEL. Langerhans cell histiocytosis cells showed strong expression of p53 and in some cases co-expression of Fas and Fas-L. The expression of Fas-L was significantly higher in infiltrates from patients with single-system disease. The actual...... number of pathological Langerhans cells in apoptosis as estimated by TUNEL was low. CONCLUSIONS: The low number of TUNEL-reactive cells can be explained by the rapid turnover of apoptotic cells in the tissue, not leaving the apoptotic cells long enough in the tissue to be detected. The co...

  3. New Nanoconfined Galvanic Replacement Synthesis of Hollow Sb@C Yolk-Shell Spheres Constituting a Stable Anode for High-Rate Li/Na-Ion Batteries.

    Science.gov (United States)

    Liu, Jun; Yu, Litao; Wu, Chao; Wen, Yuren; Yin, Kuibo; Chiang, Fu-Kuo; Hu, Renzong; Liu, Jiangwen; Sun, Litao; Gu, Lin; Maier, Joachim; Yu, Yan; Zhu, Min

    2017-02-16

    In the current research project, we have prepared a novel Sb@C nanosphere anode with biomimetic yolk-shell structure for Li/Na-ion batteries via a nanoconfined galvanic replacement route. The yolk-shell microstructure consists of Sb hollow yolk completely protected by a well-conductive carbon thin shell. The substantial void space in the these hollow Sb@C yolk-shell particles allows for the full volume expansion of inner Sb while maintaining the framework of the Sb@C anode and developing a stable SEI film on the outside carbon shell. As for Li-ion battery anode, they displayed a large specific capacity (634 mAh g(-1)), high rate capability (specific capabilities of 622, 557, 496, 439, and 384 mAh g(-1) at 100, 200, 500, 1000, and 2000 mA g(-1), respectively) and stable cycling performance (a specific capacity of 405 mAh g(-1) after long 300 cycles at 1000 mA g(-1)). As for Na-ion storage, these yolk-shell Sb@C particles also maintained a reversible capacity of approximate 280 mAh g(-1) at 1000 mA g(-1) after 200 cycles.

  4. Heritable retroviral transgenes are highly expressed in chickens.

    OpenAIRE

    Briskin, M J; Hsu, R Y; Boggs, T; Schultz, J. A.; Rishell, W; Bosselman, R A

    1991-01-01

    This report describes expression of heritable reticuloendotheliosis virus (REV) vector ME111 in 20 independent lines of transgenic chickens. The results are strikingly different from studies of Moloney virus in transgenic mice, where restricted expression of inherited proviruses has led to their use primarily as insertional mutagens rather than general agents for gene transfer. In contrast, the REV ME111 provirus is actively transcribed in a variety of tissues from transgenic chickens, is exp...

  5. Homologous high-throughput expression and purification of highly conserved E coli proteins

    Directory of Open Access Journals (Sweden)

    Duchmann Rainer

    2007-06-01

    Full Text Available Abstract Background Genetic factors and a dysregulated immune response towards commensal bacteria contribute to the pathogenesis of Inflammatory Bowel Disease (IBD. Animal models demonstrated that the normal intestinal flora is crucial for the development of intestinal inflammation. However, due to the complexity of the intestinal flora, it has been difficult to design experiments for detection of proinflammatory bacterial antigen(s involved in the pathogenesis of the disease. Several studies indicated a potential association of E. coli with IBD. In addition, T cell clones of IBD patients were shown to cross react towards antigens from different enteric bacterial species and thus likely responded to conserved bacterial antigens. We therefore chose highly conserved E. coli proteins as candidate antigens for abnormal T cell responses in IBD and used high-throughput techniques for cloning, expression and purification under native conditions of a set of 271 conserved E. coli proteins for downstream immunologic studies. Results As a standardized procedure, genes were PCR amplified and cloned into the expression vector pQTEV2 in order to express proteins N-terminally fused to a seven-histidine-tag. Initial small-scale expression and purification under native conditions by metal chelate affinity chromatography indicated that the vast majority of target proteins were purified in high yields. Targets that revealed low yields after purification probably due to weak solubility were shuttled into Gateway (Invitrogen destination vectors in order to enhance solubility by N-terminal fusion of maltose binding protein (MBP, N-utilizing substance A (NusA, or glutathione S-transferase (GST to the target protein. In addition, recombinant proteins were treated with polymyxin B coated magnetic beads in order to remove lipopolysaccharide (LPS. Thus, 73% of the targeted proteins could be expressed and purified in large-scale to give soluble proteins in the range of 500

  6. Functional validation of a constitutive autonomous silencer element.

    Directory of Open Access Journals (Sweden)

    Heyuan Qi

    Full Text Available Sequences of the genome that are capable of silencing gene expression are thought to play a key role in gene regulation. However, very few silencer elements capable of functioning in mammalian cells have been described, and only a fraction of these have been tested for the ability to function in an autonomous fashion. We report here the characterization and functional validation of a constitutive autonomous silencer element from the human genome called T39, and the comparison of T39 to three other putative silencer elements previously described by others. Functional analysis included one assay for enhancer-blocking insulator activity and two independent assays for silencer activity, all based on stable transfection and comparison to a neutral spacer control. In erythroid K562 cells, T39 exhibited potent silencer activity, the previously described element PRE2-S5 exhibited modest silencer activity, and the two other previously described elements exhibited no silencer activity. T39 was further found to be capable of silencing three disparate promoters, of silencing gene expression in three disparate cell lines, and of functioning as a single copy in a topology-independent manner. Of the four elements analyzed, only T39 exhibits a constitutive pattern of DNase hypersensitivity and binding by CTCF. In its native location the T39 element also exhibits a unique interaction profile with a subset of distal putative regulatory elements. Taken together, these studies validate T39 as a constitutive autonomous silencer, identify T39 as a defined control for future studies of other regulatory elements such as insulators, and provide a basic chromatin profile for one highly potent silencer element.

  7. Expression Analysis of miRNAs and Highly-expressed Small RNAs in Two Rice Subspecies and their Reciprocal Hybrids

    Institute of Scientific and Technical Information of China (English)

    Fangfang Chen; Guangming He; Hang He; Wei Chen; Xiaopeng Zhu; Manzhong Liang; Liangbi Chen; Xing Wang Deng

    2010-01-01

    Heterosis,or hybrid vigor,is the phenomenon whereby progeny of two inbred lines exhibit superior agronomic performance compared with either parent.We analyzed the expression of miRNAs and highly expressed small RNAs(defined according to Solexa sequencing results)in two rice(Oryza sativa)subspecies(japonica cv.Nipponbare and indica cv.93-11)and their reciprocal hybrids using microarrays.We found that of all the 1141 small RNAs tested,140(12%,140 of 1141)and 157(13%,157 of 1141)were identified being significantly differentially expressed in two reciprocal hybrids,respectively.All possible modes of action,including additive,high- and low- parent,above high- and below low-parent modes were exhibited.Both F1 hybrids showed non-additive expression patterns,with downregulation predominating.Interestingly,15 miRNAs displayed stark opposite expression trends relative to midparent in reciprocal hybrids.Computational prediction of targets of differentially expressed miRNAs showed that they participated in multifaceted developmental pathways,and were not distinguishable from the targets of non-differentially expressed miRNAs.Together,our findings reveal that small RNAs play roles in heterosis and add a new layer in the understanding and exploitation of molecular mechanisms of heterosis.

  8. DAG expression: high-throughput gene expression analysis of real-time PCR data using standard curves for relative quantification.

    Directory of Open Access Journals (Sweden)

    María Ballester

    Full Text Available BACKGROUND: Real-time quantitative PCR (qPCR is still the gold-standard technique for gene-expression quantification. Recent technological advances of this method allow for the high-throughput gene-expression analysis, without the limitations of sample space and reagent used. However, non-commercial and user-friendly software for the management and analysis of these data is not available. RESULTS: The recently developed commercial microarrays allow for the drawing of standard curves of multiple assays using the same n-fold diluted samples. Data Analysis Gene (DAG Expression software has been developed to perform high-throughput gene-expression data analysis using standard curves for relative quantification and one or multiple reference genes for sample normalization. We discuss the application of DAG Expression in the analysis of data from an experiment performed with Fluidigm technology, in which 48 genes and 115 samples were measured. Furthermore, the quality of our analysis was tested and compared with other available methods. CONCLUSIONS: DAG Expression is a freely available software that permits the automated analysis and visualization of high-throughput qPCR. A detailed manual and a demo-experiment are provided within the DAG Expression software at http://www.dagexpression.com/dage.zip.

  9. Impacts of high β-galactosidase expression on central metabolism of recombinant Pichia pastoris GS115 using glucose as sole carbon source via (13)C metabolic flux analysis.

    Science.gov (United States)

    Nie, Yongsheng; Huang, Mingzhi; Lu, Junjie; Qian, Jiangchao; Lin, Weilu; Chu, Ju; Zhuang, Yingping; Zhang, Siliang

    2014-10-10

    The yeast Pichia pastoris GS115 is a widely used microbial cell factory for the production of heterologous protein. In order to reveal the impacts of high heterologous protein expression on the central metabolism of Pichia pastoris GS115 using glucose as sole carbon source, we engineered a high β-galactosidase expression strain P. pastoris G1HL and a low expression control strain P. pastoris GHL through controlling the initiation strength of constitutive promoter pGAP. The carbon flux distributions in these two strains were quantified via (13)C metabolic flux analysis. Compared to the control strain, G1HL showed a lower growth rate, a higher flux through glycolysis pathway, a higher flux through pentose phosphate pathway, and a lower flux through by-products secretion pathway. The metabolic flux redistribution in G1HL was thought to compensate the increased redox cofactors and energy demands caused by the high protein expression. Although the fluxes through Krebs cycle in two engineered strains were almost the same, they were significantly lower than those in wild strain. The enhanced expression of β-galactosidase by glutamate supplementation demonstrated the potential of P. pastoris GS115 to catabolize more carbon through the Krebs cycle for even higher protein expression. In conclusion, our work indicates that P. pastoris GS115 can readjusts the central metabolism for higher heterologous protein expression and provides strategies for strain development or process optimization for enhancing production of heterologous protein.

  10. The Spanish Constitution, the Constitutional Court and the Catalan Referendum

    DEFF Research Database (Denmark)

    Abat Ninet, Antoni

    2015-01-01

    and politeia that Aristotle defined now clashes between two powerful symbolic and romantic phenomena. In the Spanish-Catalan binomial scenario, there are some elements that need to be analysed to obtain a complete picture of the constitutional possibilities to of accommodating a Catalan self...

  11. High expression of NPY receptors in the human testis.

    Science.gov (United States)

    Körner, Meike; Waser, Beatriche; Thalmann, George N; Reubii, Jean Claude

    2011-04-30

    NPY receptors represent novel molecular therapeutic targets in cancer and obesity. However, the extent of NPY receptor expression in normal human tissues is poorly investigated. Based on the role of NPY in reproductive functions, the NPY receptor expression was studied in 25 normal human testes and, additionally, 24 testicular tumors using NPY receptor autoradiography. In the normal testis, Leydig cells strongly expressed NPY receptor subtype Y2, and small arterial blood vessels Y1. Y2 receptors were found to be functional with agonist-stimulated [(35)S]GTPγS binding autoradiography. Full functional integrity of the NPY system was further suggested by the immunohistochemical detection of NPY peptide in nerve fibers directly adjacent to Leydig cells and arteries. Germ cell tumors expressed Y1 and Y2 on tumor cells in 33% and Y1 on intratumoral blood vessels in 50%. Based on its strong NPY receptor expression in Leydig cells and blood vessels, the normal human testis represents a potentially important physiological and pharmalogical NPY target.

  12. Time and constitution of sense

    Directory of Open Access Journals (Sweden)

    Pedro Gerardo Acosta

    2014-06-01

    Full Text Available This article proposes a reflection over our time-consciousness under the Phenomenology of Edmund Husserl. The idea is make a release the key role of the sense constitution like the fundament and development of the ongoing intentionality, a shape that make the possibility to catch sight of the sense of every life situation like conscience experience that displays itself over the time, and open the world of the Phenomenon World, constituted in the flux and flow of our live experience. The immanent time in which the things served in a lived-present inevitably displays to its own immediate-past of retentions, then of commemorations, constituting and enabling, not just the sense of ever present, but the sense of our own past like memory and our future like expectative. This reflection is based and supporter over the text “Phenomenology Lesson of the Internal Time-Consiusness” (Husserl, 2002.

  13. Synthesis and high expression of chitin deacetylase from Colletotrichum lindemuthianum in Pichia pastoris GS115.

    Science.gov (United States)

    Kang, Lixin; Chen, Xiaomei; Zhai, Chao; Ma, Lixin

    2012-09-01

    A gene, ClCDA, encoding chitin deacetylase from Colletotrichum lindemuthianum, was optimized according to the codon usage bias of Pichia pastoris and synthesized in vitro by overlap extension PCR. It was secretorily expressed in P. pastoris GS115 using the constitutive expression vector pHMB905A. The expression level reached the highest with 110 mg/l culture supernatant after 72 h of methanol induction, which comprised 77.27 U/mg chitin deacetylase activity. SDS-PAGE, mass spectrometry, and deglycosylation assays demonstrated that partial recombinant protein was glycosylated with an apparent molecular mass of 33 kDa. The amino acid sequences of recombinant proteins were confirmed by mass spectrometry.

  14. Constitutional Analysis of Intellectual Property

    Directory of Open Access Journals (Sweden)

    AJ van der Walt

    2014-04-01

    Full Text Available This article analyses the Constitutional Court’s treatment of property interests in the face of state regulation to gain an understanding of the type of state interference that is justifiable in terms of section 25(1 of the Bill of Rights. This is done by examining the Constitutional Court’s dicta relating to the meaning of deprivation and how these inform the meaning of property in the constitutional context. The methodology that the Constitutional Court has formulated to assess if state interference complies with the provisions of section 25 is explained to show the type of state regulation that has been found legitimate. We then consider how this understanding of constitutional property and the state’s legitimate exercise of its inherent police power interact in the setting of intellectual property by contrasting the various policy objectives underlying the different statutory regimes governing intellectual property. This theoretical analysis is then applied to two contemporary examples of feasible state interference with existing intellectual property interests, namely the proposed plain packaging measures which severely restrict the use of tobacco trade marks, and a fair dealing exception allowing the use of copyright works for the purpose of parody. These examples serve to illustrate the context and manner in which intellectual property interests may come before the Court and the necessary differentiation with which these interests should be treated. The appropriate judicial assessment of the true impact that state action could have on vested property interests is explained and contrasted with the balancing exercise that is employed at the earlier stage of policy making. This discussion is concluded by highlighting some of the interpretational issues that will arise and how some constitutional values could be curtailed in the absence of legislative intervention.

  15. Recombinant cells that highly express chromosomally-integrated heterologous gene

    Science.gov (United States)

    Ingram, Lonnie O.; Ohta, Kazuyoshi; Wood, Brent E.

    2007-03-20

    Recombinant host cells are obtained that comprise (A) a heterologous, polypeptide-encoding polynucleotide segment, stably integrated into a chromosome, which is under transcriptional control of an endogenous promoter and (B) a mutation that effects increased expression of the heterologous segment, resulting in enhanced production by the host cells of each polypeptide encoded by that segment, relative to production of each polypeptide by the host cells in the absence of the mutation. The increased expression thus achieved is retained in the absence of conditions that select for cells displaying such increased expression. When the integrated segment comprises, for example, ethanol-production genes from an efficient ethanol producer like Zymomonas mobilis, recombinant Escherichia coli and other enteric bacterial cells within the present invention are capable of converting a wide range of biomass-derived sugars efficiently to ethanol.

  16. A Communicatively Constituted Online Crisis

    DEFF Research Database (Denmark)

    Valentini, Chiara; Romenti, Stefania; Kruckeberg, Dean

    2017-01-01

    mediated, then online crises and their meanings are communicatively constructed. The chapter outlines how online conversations constitute discursive practices that make publics aware of critical situations of which they may or may not know and can even construct reputational crises. The chapter concludes...... by offering suggestions on how to study online critical conversations through the lens of a communicative constitution perspective that could inform how critical issues eventually transform and become crises and how crisis perceptions evolve and are discursively shaped by communicative practices occurring...

  17. Constitutive expression of CaXTH3, a hot pepper xyloglucan endotransglucosylase/hydrolase, enhanced tolerance to salt and drought stresses without phenotypic defects in tomato plants (Solanum lycopersicum cv. Dotaerang).

    Science.gov (United States)

    Choi, Jun Young; Seo, Young Sam; Kim, Su Jin; Kim, Woo Taek; Shin, Jeong Sheop

    2011-05-01

    The hot pepper xyloglucan endo-trans-gluco-sylase/hydrolase (CaXTH3) gene that was inducible by a broad spectrum of abiotic stresses in hot pepper has been reported to enhance tolerance to drought and high salinity in transgenic Arabidopsis. To assess whether CaXTH3 is a practically useful target gene for improving the stress tolerance of crop plants, we ectopically over-expressed the full-length CaXTH3 cDNA in tomato (Solanum lycopersicum cv. Dotaerang) and found that the 35S:CaXTH3 transgenic tomato plants exhibited a markedly increased tolerance to salt and drought stresses. Transgenic tomato plants exposed to a salt stress of 100 mM NaCl retained the chlorophyll in their leaves and showed normal root elongation. They also remained green and unwithered following exposure to 2 weeks of dehydration. A high proportion of stomatal closures in 35S:CaXTH3 was likely to be conferred by increased cell-wall remodeling activity of CaXTH3 in guard cell, which may reduce transpirational water loss in response to dehydration stress. Despite this increased stress tolerance, the transgenic tomato plants showed no detectable phenotype defects, such as abnormal morphology and growth retardation, under normal growth conditions. These results raise the possibility that CaXTH3 gene is appropriate for application in genetic engineering strategies aimed at improving abiotic stress tolerance in agriculturally and economically valuable crop plants.

  18. A Highly Efficient Gene Expression Programming (GEP Model for Auxiliary Diagnosis of Small Cell Lung Cancer.

    Directory of Open Access Journals (Sweden)

    Zhuang Yu

    Full Text Available Lung cancer is an important and common cancer that constitutes a major public health problem, but early detection of small cell lung cancer can significantly improve the survival rate of cancer patients. A number of serum biomarkers have been used in the diagnosis of lung cancers; however, they exhibit low sensitivity and specificity.We used biochemical methods to measure blood levels of lactate dehydrogenase (LDH, C-reactive protein (CRP, Na+, Cl-, carcino-embryonic antigen (CEA, and neuron specific enolase (NSE in 145 small cell lung cancer (SCLC patients and 155 non-small cell lung cancer and 155 normal controls. A gene expression programming (GEP model and Receiver Operating Characteristic (ROC curves incorporating these biomarkers was developed for the auxiliary diagnosis of SCLC.After appropriate modification of the parameters, the GEP model was initially set up based on a training set of 115 SCLC patients and 125 normal controls for GEP model generation. Then the GEP was applied to the remaining 60 subjects (the test set for model validation. GEP successfully discriminated 281 out of 300 cases, showing a correct classification rate for lung cancer patients of 93.75% (225/240 and 93.33% (56/60 for the training and test sets, respectively. Another GEP model incorporating four biomarkers, including CEA, NSE, LDH, and CRP, exhibited slightly lower detection sensitivity than the GEP model, including six biomarkers. We repeat the models on artificial neural network (ANN, and our results showed that the accuracy of GEP models were higher than that in ANN. GEP model incorporating six serum biomarkers performed by NSCLC patients and normal controls showed low accuracy than SCLC patients and was enough to prove that the GEP model is suitable for the SCLC patients.We have developed a GEP model with high sensitivity and specificity for the auxiliary diagnosis of SCLC. This GEP model has the potential for the wide use for detection of SCLC in less

  19. Identification of several soybean cytosolic glutamine synthetase transcripts highly or specifically expressed in nodules: expression studies using one of the corresponding genes in transgenic Lotus corniculatus.

    Science.gov (United States)

    Marsolier, M C; Debrosses, G; Hirel, B

    1995-01-01

    A DNA fragment containing sequences hybridizing to the 5' region of GS15, a gene encoding soybean cytosolic glutamine synthetase, was isolated from a soybean genomic library. Mapping and partial sequence analysis of the genomic clone revealed that it encodes a cytosolic GS gene, GS21, which is different from GS15. In parallel, a number of cDNA clones encoding cytosolic GS were isolated using the coding region of pGS20 as a probe (pGS20 is a cDNA clone which corresponds to a transcript of the GS15 gene). Two new full-length cDNAs designated pGS34 and pGS38 were isolated and sequenced. In the 5' non-coding region a strong homology was found between the two clones and the GS21 gene. However, none of these sequences were identical, which suggests that there are at least three members in this group of genes. In order to determine their relative levels of transcription, specific sequences from pGS34, pGS38 and GS21 were used in an RNAse protection assay. This experiment clearly showed that GS21 and the gene encoding pGS38 are specifically expressed in young or mature nodules, whereas the gene encoding pGS34 is highly transcribed in nodules and constitutively expressed at a lower level in other soybean organs. In order to further analyse the molecular mechanisms controlling GS21 transcription, different fragments of the promoter region were fused to the Escherichia coli reporter gene encoding beta-glucuronidase (GUS) and the constructs were introduced into Lotus corniculatus via Agrobacterium rhizogenes-mediated transformation. Analysis of GUS activity showed that the GS21 promoter-GUS constructs were expressed in the vasculature of all vegetative organs. This result is discussed in relation to species-specific metabolic and developmental characteristics of soybean and Lotus.

  20. New Constitutionalism for Biosiversity vs. Neoconstitutionalism of Risk

    Directory of Open Access Journals (Sweden)

    Michele Carducci

    2016-08-01

    Full Text Available http://dx.doi.org/10.5007/2177-7055.2016v37n73p255 Based on an “eco-systemic” democracy that seeks to preserve biodiversity through the recognition of the co-evolutionary link between nature and culture, the Andean Constitutionalism emerges as the expression of a counter-hegemonic constitutionalism committed to the construction of a new institutional framework through the inclusion of new participatory and intercultural mechanisms. Departing from western constitutional paradigms, this groundbreaking constitutionalism revisits the “Gaia hypothesis” and legitimizes a real “social contract” among the people and nature, and instead of considering it as an “object” of ownership, exploitation, or conservation, it regards nature as a legal “subject” and primary source of society itself and the Constitution as its “legal grantor and protector”.

  1. Constitutive relations in optics in terms of geometric algebra

    Science.gov (United States)

    Dargys, A.

    2015-11-01

    To analyze the electromagnetic wave propagation in a medium the Maxwell equations should be supplemented by constitutive relations. At present the classification of linear constitutive relations is well established in tensorial-matrix and exterior p-form calculus. Here the constitutive relations are found in the context of Clifford geometric algebra. For this purpose Cl1,3 algebra that conforms with relativistic 4D Minkowskian spacetime is used. It is shown that the classification of linear optical phenomena with the help of constitutive relations in this case comes from the structure of Cl1,3 algebra itself. Concrete expressions for constitutive relations which follow from this algebra are presented. They can be applied in calculating the propagation properties of electromagnetic waves in any anisotropic, linear and nondissipative medium.

  2. Conjugated Linoleic Acid Supplementation under a High-Fat Diet Modulates Stomach Protein Expression and Intestinal Microbiota in Adult Mice.

    Science.gov (United States)

    Chaplin, Alice; Parra, Pilar; Serra, Francisca; Palou, Andreu

    2015-01-01

    The gastrointestinal tract constitutes a physiological interface integrating nutrient and microbiota-host metabolism. Conjugated linoleic acids (CLA) have been reported to contribute to decreased body weight and fat accretion. The modulation by dietary CLA of stomach proteins related to energy homeostasis or microbiota may be involved, although this has not been previously analysed. This is examined in the present study, which aims to underline the potential mechanisms of CLA which contribute to body weight regulation. Adult mice were fed either a normal fat (NF, 12% kJ content as fat) or a high-fat (HF, 43% kJ content as fat) diet. In the latter case, half of the animals received daily oral supplementation of CLA. Expression and content of stomach proteins and specific bacterial populations from caecum were analysed. CLA supplementation was associated with an increase in stomach protein expression, and exerted a prebiotic action on both Bacteroidetes/Prevotella and Akkermansia muciniphila. However, CLA supplementation was not able to override the negative effects of HF diet on Bifidobacterium spp., which was decreased in both HF and HF+CLA groups. Our data show that CLA are able to modulate stomach protein expression and exert a prebiotic effect on specific gut bacterial species.

  3. Towards a global climate constitution

    NARCIS (Netherlands)

    Weikard, H.P.

    2011-01-01

    In this paper my concern is the study of the incentives of individual countries to sign an international climate agreement that sets the terms of a climate constitution, that is, it establishes emission rights and rules for trading these rights to combat the climate problem effectively and efficient

  4. The Constitutive Power of Twitter

    DEFF Research Database (Denmark)

    Albu, Oana Brindusa; Etter, Michael Andreas

    organizations. The analysis illustrates how specific Twitter interactions, i.e., hashtags, become hypertexts—a type of authoritative texts—which simultaneously constitute an organizational actor or act as a pastiche of it. The study contributes to extant research by illustrating how hypertextuality...

  5. Constitutional Change Marks Ideological Breakthrough

    Institute of Scientific and Technical Information of China (English)

    2004-01-01

    The amendments to the Constitution of the People'sRepublic of China, the fourth since the constitutionwas promulgated in 1982 and proposed by theCentral Committee of the Communist Party of China(CPC), are expected to enshrine the respect andprotection of human rights, the protection of private

  6. Take Advantage of Constitution Day

    Science.gov (United States)

    McCune, Bonnie F.

    2008-01-01

    The announcement of the mandate for Constitution and Citizenship Day shortly before September, 2005, probably led to groans of dismay. Not another "must-do" for teachers and schools already stressed by federal and state requirements for standardized tests, increasingly rigid curricula, and scrutiny from the public and officials. But the…

  7. A Constitutive Model for Isothermal Pseudoelasticity Coupled with Plasticity

    Science.gov (United States)

    Jiang, Dongjie; Landis, Chad M.

    2016-12-01

    In this paper, a new constitutive model for isothermal pseudoelastic shape memory alloys is presented. The model is based upon a kinematic hardening framework that was previously developed for ferroelastic and ferroelectric switching behavior. The basis of the model includes a transformation surface, an associated flow rule for transformation strain, and kinematic hardening with the back stresses represented by a transformation potential that is dependent upon the transformation strain. In contrast to many models that introduce tension/compression asymmetry by devising transformation surfaces in terms of invariants of the stress tensor, this model achieves this capability by means of expressing the transformation potential from which the back stresses are derived as a weighted mix of two potentials that are, respectively, calibrated to measured tensile and compressive responses. Additionally, in this model, plastic deformation is allowed to occur at high stresses by employing a standard J2-based yield surface with isotropic hardening. Finally, to demonstrate the ability of the constitutive model to perform in highly non-proportional loading states, some finite element simulations on crack tip fields are presented.

  8. Ideology, constitutional culture and institutional change: the EU constitution as reflection of Europe’s emergent postmodernism

    OpenAIRE

    Nikolay Wenzel

    2007-01-01

    Using the example of the European Constitution, this paper argues that ideology plays a much more important role in institutional change than has been indicated hitherto in the literature. Rather than being an intellectual parlor-game, Postmodernism has emerged through European high culture to find its voice in the new Constitution. Although it was rejected by a critical mass of voters, the proposed Constitution offers a telling glimpse into the European intellectual mindset – especially sinc...

  9. Constitutive Relation of Yunjialing Anthracite Under Medium Strain Rate

    Institute of Scientific and Technical Information of China (English)

    GAO Wen-jiao; SHAN Ren-liang; WANG Gong-cheng; CHENG Rui-qiang

    2007-01-01

    By means of the split Hopkinson pressure bar (SHPB) testing system, this paper presents a dynamic constitutive relation of anthracite at a strain rate of ε =5-85s-1. Generally, the dynamic stress-strain curve for this kind of anthracite under uni-axial compression has the following four stages: a non-linear loading stage, a plastic yielding stage, a strain-strengthening stage and an unloading breakage stage. Correspondingly, the initial elastic modulus Eb, the yielding strength σs and the ultimate strength σb increase along with an increasing strain rate. The time-dependent elasticity was identified when we analyzed the mechanical properties of anthracite. Based on characteristics of measured dynamic stress-strain curves and an analysis of existing rock dynamic constitutive models, as well as a preparatory simulation, a new visco-elastic damage model has been introduced in this paper. A linear spring is put parallel to two Maxwell units with different relaxation times to express two distinct plastic flows. The damage D is equal to [Eb- E(εi)]/Eb, where Eb is the beginning modulus and the E(εi) is the slope of a connected line between the origin point and any other point on a tested stress-strain curve. In the new constitutive model, one Maxwell unit with low relaxation time (ψ)1 is used to describe the response of anthracite to a low strain rate, while the other, with a high relaxation time (ψ)2 describes the response of anthracite to a high strain rate. Simulated stress-strain curves from the new model are consistent with the measured curves.

  10. High levels of protein expression using different mammalian CMV promoters in several cell lines.

    Science.gov (United States)

    Xia, Wei; Bringmann, Peter; McClary, John; Jones, Patrick P; Manzana, Warren; Zhu, Ying; Wang, Soujuan; Liu, Yi; Harvey, Susan; Madlansacay, Mary Rose; McLean, Kirk; Rosser, Mary P; MacRobbie, Jean; Olsen, Catherine L; Cobb, Ronald R

    2006-01-01

    With the recent completion of the human genome sequencing project, scientists are faced with the daunting challenge of deciphering the function of these newly found genes quickly and efficiently. Equally as important is to produce milligram quantities of the therapeutically relevant gene products as quickly as possible. Mammalian expression systems provide many advantages to aid in this task. Mammalian cell lines have the capacity for proper post-translational modifications including proper protein folding and glycosylation. In response to the needs described above, we investigated the protein expression levels driven by the human CMV in the presence or absence of intron A, the mouse and rat CMV promoters with intron A, and the MPSV promoter in plasmid expression vectors. We evaluated the different promoters using an in-house plasmid vector backbone. The protein expression levels of four genes of interest driven by these promoters were evaluated in HEK293EBNA and CHO-K1 cells. Stable and transient transfected cells were utilized. In general, the full-length human CMV, in the presence of intron A, gave the highest levels of protein expression in transient transfections in both cell lines. However, the MPSV promoter resulted in the highest levels of stable protein expression in CHO-K1 cells. Using the CMV driven constitutive promoters in the presence of intron A, we have been able to generate >10 microg/ml of recombinant protein using transient transfections.

  11. Tumor endothelial cells express high pentraxin 3 levels.

    Science.gov (United States)

    Hida, Kyoko; Maishi, Nako; Kawamoto, Taisuke; Akiyama, Kosuke; Ohga, Noritaka; Hida, Yasuhiro; Yamada, Kenji; Hojo, Takayuki; Kikuchi, Hiroshi; Sato, Masumi; Torii, Chisaho; Shinohara, Nobuo; Shindoh, Masanobu

    2016-12-01

    It has been described that tumor progression has many similarities to inflammation and wound healing in terms of the signaling processes involved. Among biological responses, angiogenesis, which is necessary for tumor progression and metastasis, is a common hallmark; therefore, tumor blood vessels have been considered as important therapeutic targets in anticancer therapy. We focused on pentraxin 3 (PTX3), which is a marker of cancer-related inflammation, but we found no reports on its expression and function in tumor blood vessels. Here we showed that PTX3 is expressed in mouse and human tumor blood vessels based on immunohistochemical analysis. We found that PTX3 is upregulated in primary mouse and human tumor endothelial cells compared to normal endothelial cells. We also showed that PTX3 plays an important role in the proliferation of the tumor endothelial cells. These results suggest that PTX3 is an important target for antiangiogenic therapy.

  12. A study on peace principle of the constitution of Japan

    OpenAIRE

    1984-01-01

    In 1930s and 40s when I was a student and then a naval officer, whether Japan could offer any political principle for international society or not was an agonizing question to me. Apparently, the Constitution of Great Empire of Japan of 1889 did not have any, or rather it was forcing Japanese nation to be alienated from the world.The Constitution of Japan of 1946 is often called a 'peace constitution', having the sentences in paragraph 2 of its Preamble, which express pacifistic intention, an...

  13. Neural Network Model for the Constitutive Relations of Soil

    Institute of Scientific and Technical Information of China (English)

    Zeng Jing; Wang Jing-tao

    2003-01-01

    The soil constitutive relation is one of the important issues in soil mechanics. It is very difficult to establish mathematical models because of the complexity of soil mechanical behavior. We propose a new method of neural network analysis for establishing soil constitutive models. Based on triaxial experiments of sand in the laboratory, the nonlinear constitutive models of sand expressed by the neural network were set up. In comparison with Duncan-Chang's model, the neural network method for sand modeling has been proved to be more convenient, accurate and it has a strong fault-tolerance function.

  14. Incorporating the Johnson-Cook Constitutive Model and a Soft Computational Approach for Predicting the High-Temperature Flow Behavior of Sn-5Sb Solder Alloy: A Comparative Study for Processing Map Development

    Science.gov (United States)

    Vafaeenezhad, H.; Seyedein, S. H.; Aboutalebi, M. R.; Eivani, A. R.

    2016-09-01

    The high-temperature flow behavior of Sn-5Sb lead-free solder alloy has been investigated using isothermal hot compression experiments at 298 K to 400 K and strain rate between 0.0005 s-1 and 0.01 s-1. The flow stress under these test conditions was modeled using constitutive equations based on the Johnson-Cook (J-C) model and an artificial neural network (ANN). Three input factors, i.e., temperature, strain rate, and true strain, were incorporated into the network, and the flow stress was considered as the system output. One hidden layer was adopted in the simulations. Furthermore, a comparative study was carried out on the potential of the two proposed models to characterize the high-temperature flow behavior of this alloy. The capability of the models was assessed by comparing the simulation predictions using a correlation coefficient (R 2). The stresses predicted by both models presented good agreement with experimental results. In addition, it was found that the ANN model could predict the high-temperature deformation more precisely over the whole temperature and strain rate ranges. However, this is strongly dependent on the availability of extensive, high-quality data and characteristic variables.

  15. Incorporating the Johnson-Cook Constitutive Model and a Soft Computational Approach for Predicting the High-Temperature Flow Behavior of Sn-5Sb Solder Alloy: A Comparative Study for Processing Map Development

    Science.gov (United States)

    Vafaeenezhad, H.; Seyedein, S. H.; Aboutalebi, M. R.; Eivani, A. R.

    2017-01-01

    The high-temperature flow behavior of Sn-5Sb lead-free solder alloy has been investigated using isothermal hot compression experiments at 298 K to 400 K and strain rate between 0.0005 s-1 and 0.01 s-1. The flow stress under these test conditions was modeled using constitutive equations based on the Johnson-Cook (J-C) model and an artificial neural network (ANN). Three input factors, i.e., temperature, strain rate, and true strain, were incorporated into the network, and the flow stress was considered as the system output. One hidden layer was adopted in the simulations. Furthermore, a comparative study was carried out on the potential of the two proposed models to characterize the high-temperature flow behavior of this alloy. The capability of the models was assessed by comparing the simulation predictions using a correlation coefficient ( R 2). The stresses predicted by both models presented good agreement with experimental results. In addition, it was found that the ANN model could predict the high-temperature deformation more precisely over the whole temperature and strain rate ranges. However, this is strongly dependent on the availability of extensive, high-quality data and characteristic variables.

  16. Macroscopic Expressions of Molecular Adiabatic Compressibility of Methyl and Ethyl Caprate under High Pressure and High Temperature

    Directory of Open Access Journals (Sweden)

    Fuxi Shi

    2014-01-01

    Full Text Available The molecular compressibility, which is a macroscopic quantity to reveal the microcompressibility by additivity of molecular constitutions, is considered as a fixed value for specific organic liquids. In this study, we introduced two calculated expressions of molecular adiabatic compressibility to demonstrate its pressure and temperature dependency. The first one was developed from Wada’s constant expression based on experimental data of density and sound velocity. Secondly, by introducing the 2D fitting expressions and their partial derivative of pressure and temperature, molecular compressibility dependency was analyzed further, and a 3D fitting expression was obtained from the calculated data of the first one. The third was derived with introducing the pressure and temperature correction factors based on analogy to Lennard-Jones potential function and energy equipartition theorem. In wide range of temperatures (293

  17. Non-coplanar polychlorinated biphenyls (PCBs) are direct agonists for the human pregnane-X receptor and constitutive androstane receptor, and activate target gene expression in a tissue-specific manner

    Energy Technology Data Exchange (ETDEWEB)

    Al-Salman, Fadheela; Plant, Nick, E-mail: N.Plant@Surrey.ac.uk

    2012-08-15

    The polychlorinated biphenyl group possesses high environmental persistence, leading to bioaccumulation and a number of adverse effects in mammals. Whilst coplanar PCBs elicit their toxic effects through agonism of the aryl hydrocarbon receptor; however, non-coplanar PCBs are not ligands for AhR, but may be ligands for members of the nuclear receptor family of proteins. To better understand the biological actions of non-coplanar PCBs, we have undertaken a systematic analysis of their ability to activate PXR and CAR-mediated effects. Cells were exposed to a range of non-coplanar PCBs (99, 138, 153, 180 and 194), or the coplanar PCB77: Direct activation of PXR and CAR was measured using a mammalian receptor activation assay in human liver cells, with rifampicin and CITCO used as positive controls ligands for PXR and CAR, respectively; activation of target gene expression was examined using reporter gene plasmids for CYP3A4 and MDR1 transfected into liver, intestine and lung cell lines. Several of the non-coplanar PCBs directly activated PXR and CAR, whilst the coplanar PCB77 did not. Non-coplanar PCBs were also able to activate PXR/CAR target gene expression in a substitution- and tissue-specific manner. Non-coplanar PCBs act as direct activators for the nuclear receptors PXR and CAR, and are able to elicit transcriptional activation of target genes in a substitution- and tissue-dependent manner. Chronic activation of PXR/CAR is linked to adverse effects and must be included in any risk assessment of PCBs. -- Highlights: ► Several Non-coplanar PCBs are able to directly activate both PXR and CAR in vitro. ► PCB153 is the most potent direct activator of PXR and CAR nuclear receptors. ► Non-coplanar PCB activation of CYP3A4/MDR1 reporter genes is structure-dependent. ► Non-coplanar PCB activate CYP3A4/MDR1 reporter genes in a tissue-dependent. ► PCB153 is the most potent activator of PXR/CAR target gene in all tissues.

  18. Skp2 expression is associated with high risk and elevated Ki67 expression in gastrointestinal stromal tumours

    Directory of Open Access Journals (Sweden)

    Terracciano Luigi

    2008-05-01

    Full Text Available Abstract Background Gastrointestinal stromal tumors (GIST exhibit an unpredictable clinical course and can rapidly progress to lethality. Predictions about the biological behavior of GIST are based on a number of canonical clinical and pathologic parameters whose validity in distinguishing between a benign and a malignant tumour is still imperfect. The aim of our study was to investigate the role of morphologic parameters and expression of cells cycle regulators as prognosticators in GIST. Methods We performed an immunohistochemical analysis for Ki67, p27Kip1, Jab1, and Skp2, on a Tissue Microarray (TMA containing 94 GIST. Expression of the above proteins was correlated to classically used prognosticators, as well as to risk groups. Clinical significance of histologic and immunohistochemical features were evaluated in 59 patients for whom follow-up information was available. Results Overexpression of Ki67 and Skp2, and p27Kip1 loss directly correlated with the high risk group (p = 0.03 for Ki67 and Skp2, p = 0.05 for p27Kip1. Jab1 expression did not exhibit correlation with risk. In 59 cases provided with clinical follow-up, high cellularity, presence of necrosis, and Ki67 overexpression were predictive of a reduced overall survival in a univariate model. The same parameters, as well as mitotic rate, tumour size, and p27Kip1 loss were indicative of a shortened relapse free survival interval. High cellularity, and high mitotic rate retained their prognostic significance by multivariate analysis. Conclusion Our data suggest that a number of histologic parameters in combination with immunohistochemical expression of cell cycle regulators can facilitate risk categorization and predict biologic behavior in GIST. Importantly this study demonstrates, for the first time, that Skp2 expression correlates with Ki67 expression and high risk in GIST.

  19. Combinatorial Screening for Transgenic Yeasts with High Cellulase Activities in Combination with a Tunable Expression System.

    Directory of Open Access Journals (Sweden)

    Yoichiro Ito

    Full Text Available Combinatorial screening used together with a broad library of gene expression cassettes is expected to produce a powerful tool for the optimization of the simultaneous expression of multiple enzymes. Recently, we proposed a highly tunable protein expression system that utilized multiple genome-integrated target genes to fine-tune enzyme expression in yeast cells. This tunable system included a library of expression cassettes each composed of three gene-expression control elements that in different combinations produced a wide range of protein expression levels. In this study, four gene expression cassettes with graded protein expression levels were applied to the expression of three cellulases: cellobiohydrolase 1, cellobiohydrolase 2, and endoglucanase 2. After combinatorial screening for transgenic yeasts simultaneously secreting these three cellulases, we obtained strains with higher cellulase expressions than a strain harboring three cellulase-expression constructs within one high-performance gene expression cassette. These results show that our method will be of broad use throughout the field of metabolic engineering.

  20. High Frequency Electromagnetic Field Induces Lipocalin 2 Expression in

    Directory of Open Access Journals (Sweden)

    Amaneh Mohammadi Roushandeh

    2010-06-01

    Full Text Available Objective(sNeutrophil gelatinase-associated lipocalin (NGAL/Lcn2, comprise a group of small extracellular proteins with a common β-sheet-dominated 3-dimensional structure. In the past, it was assumed that the predominant role of lipocalin was acting as transport proteins. Recently it has been found that oxidative stress induces Lcn2 expression. It has been also proved that electromagnetic field (EMF produces reactive oxygen species (ROS in different tissues. Expression of Lcn2 following exposure to electromagnetic field has been investigated in this study. Materials and MethodsBalb/c mice (8 weeks old were exposed to 3 mT, 50 HZ EMF for 2 months, 4 hr/day. Afterwards, the mice were sacrificed by cervical dislocation and livers were removed. The liver specimens were stained with Haematoxylin- Eosin (H&E and analyzed under an optical microscope. Total RNA was extracted from liver and reverse transcription was performed by SuperScript III reverse transcriptase with 1 µg of total RNA. Assessment of Lcn2 expression was performed by semiquantitative and real time- PCR.ResultsThe light microscopic studies revealed that the number of lymphocyte cells was increased compared to control and dilation of sinosoids was observed in the liver. Lcn2 was up-regulated in the mice exposed to EMF both in mRNA and protein levels.ConclusionTo the extent of our knowledge, this is the first report dealing with up-regulation of Lcn2 in liver after exposure to EMF. The up-regulation might be a compensatory response that involves cell defense pathways and protective effects against ROS. However, further and complementary studies are required in this regards.

  1. Methods of Writing Constitutional Formulas

    National Research Council Canada - National Science Library

    Raos, N; Miličević, A

    2012-01-01

    ... – expressiveness and economy, i.e. they have to express the maximal meaning with minimal means. Besides, chemical formula, being a scientific notation, has not to convey vague and scientifically unapproved meanings...

  2. Interruptions in gene expression drive highly expressed operons to the leading strand of DNA replication.

    Science.gov (United States)

    Price, Morgan N; Alm, Eric J; Arkin, Adam P

    2005-01-01

    In bacteria, most genes are on the leading strand of replication, a phenomenon attributed to collisions between the DNA and RNA polymerases. In Escherichia coli, these collisions slow the movement of the replication fork through actively transcribed genes only if they are coded on the lagging strand. For genes on both strands, however, these collisions sever nascent transcripts and interrupt gene expression. Based on these observations, we propose a new theory to explain strand bias: genes whose expression is important for fitness are selected to the leading strand because this reduces the duration of these interruptions. Our theory predicts that multi-gene operons, which are subject to longer interruptions, should be more strongly selected to the leading strand than singleton transcripts. We show that this is true even after controlling for the tendency for essential genes, which are strongly biased to the leading strand, to occur in operons. Our theory also predicts that other factors that are associated with strand bias should have stronger effects for genes that are in operons. We find that expression level and phylogenetic ubiquity are correlated with strand bias for both essential and non-essential genes, but only for genes in operons.

  3. Valuation In The Constitutional Era

    Directory of Open Access Journals (Sweden)

    Elmien du Plessis

    2015-12-01

    Full Text Available The Constitution brought about a new compensation regime for expropriations. Compensation for expropriation must now be "just and equitable". Whereas before the Constitution came into force market value played a central role in compensation for expropriation, market value is now only one factor or aspect of compensation that the court needs to take into account. Yet we find that courts tend to focus on market value and to still employ the valuation methods used to calculate market value. This article argues that the methods used to calculate the market value, once thought to be objective, are not as objective as was believed. While it is impossible to give judges specific tools for the assessment of market value, this article provides guidelines on how the calculation of compensation should be approached.

  4. Engineering of core promoter regions enables the construction of constitutive and inducible promoters in Halomonas sp.

    Science.gov (United States)

    Li, Tingting; Li, Teng; Ji, Weiyue; Wang, Qiuyue; Zhang, Haoqian; Chen, Guo-Qiang; Lou, Chunbo; Ouyang, Qi

    2016-02-01

    Halomonas strain TD01, a newly identified halophilic bacterium, has proven to be a promising low-cost host for the production of chemicals. However, genetic manipulation in Halomonas sp. is still difficult due to the lack of well-characterized and tunable expression systems. In this study, a systematic, efficient method was exploited to construct both a constitutive promoter library and inducible promoters. Porin, a highly expressed protein in Halomonas TD01, was first identified from the Halomonas TD01 proteome. Subsequent study of the intergenic region upstream of porin led to the identification of a core promoter region, including -10 and -35 elements. By randomizing the sequence between the -35 and -10 elements, a constitutive promoter library was obtained with 310-fold variation in transcriptional activity; an inducible promoter with a >200-fold induction was built by integrating a lac operator into the core promoter region. As two complementary expression systems, the constitutive and inducible promoters were then employed to regulate the biosynthetic pathway of poly-3-hydroxybutyrate (PHB) in Halomonas TD01, demonstrating the usefulness of the expression systems, furthermore, they could be applied in future metabolic engineering of Halomonas TD strains, and the systematic method used in this study can be generalized to other less-characterized bacterial strains.

  5. Telomeric expression sites are highly conserved in Trypanosoma brucei.

    Directory of Open Access Journals (Sweden)

    Christiane Hertz-Fowler

    Full Text Available Subtelomeric regions are often under-represented in genome sequences of eukaryotes. One of the best known examples of the use of telomere proximity for adaptive purposes are the bloodstream expression sites (BESs of the African trypanosome Trypanosoma brucei. To enhance our understanding of BES structure and function in host adaptation and immune evasion, the BES repertoire from the Lister 427 strain of T. brucei were independently tagged and sequenced. BESs are polymorphic in size and structure but reveal a surprisingly conserved architecture in the context of extensive recombination. Very small BESs do exist and many functioning BESs do not contain the full complement of expression site associated genes (ESAGs. The consequences of duplicated or missing ESAGs, including ESAG9, a newly named ESAG12, and additional variant surface glycoprotein genes (VSGs were evaluated by functional assays after BESs were tagged with a drug-resistance gene. Phylogenetic analysis of constituent ESAG families suggests that BESs are sequence mosaics and that extensive recombination has shaped the evolution of the BES repertoire. This work opens important perspectives in understanding the molecular mechanisms of antigenic variation, a widely used strategy for immune evasion in pathogens, and telomere biology.

  6. Enzyme free cloning for high throughput gene cloning and expression

    NARCIS (Netherlands)

    de Jong, R.N.; Daniëls, M.; Kaptein, R.; Folkers, G.E.

    2006-01-01

    Structural and functional genomics initiatives significantly improved cloning methods over the past few years. Although recombinational cloning is highly efficient, its costs urged us to search for an alternative high throughput (HTP) cloning method. We implemented a modified Enzyme Free Cloning (EF

  7. Enzyme free cloning for high throughput gene cloning and expression

    NARCIS (Netherlands)

    de Jong, R.N.; Daniëls, M.; Kaptein, R.; Folkers, G.E.

    2006-01-01

    Structural and functional genomics initiatives significantly improved cloning methods over the past few years. Although recombinational cloning is highly efficient, its costs urged us to search for an alternative high throughput (HTP) cloning method. We implemented a modified Enzyme Free Cloning

  8. Enzyme free cloning for high throughput gene cloning and expression

    NARCIS (Netherlands)

    de Jong, R.N.; Daniëls, M.; Kaptein, R.; Folkers, G.E.

    2006-01-01

    Structural and functional genomics initiatives significantly improved cloning methods over the past few years. Although recombinational cloning is highly efficient, its costs urged us to search for an alternative high throughput (HTP) cloning method. We implemented a modified Enzyme Free Cloning (EF

  9. 6082-T6高强铝合金材料本构模型试验研究%Experimental study on constitutive model of high-strength aluminum alloy 6082-T6

    Institute of Scientific and Technical Information of China (English)

    王誉瑾; 范峰; 钱宏亮; 翟希梅

    2013-01-01

    铝合金材料本构关系是铝合金结构分析和应用的基本问题之一,针对国产高强新牌号铝合金6082-T6的材料本构模型进行研究.对取自3种截面形式结构用6082-T6高强铝合金型材的90个材性试样进行了拉伸试验,统计规定非比例延伸强度、抗拉强度、弹性模量、断裂伸长率、泊松比等材料力学参数.分析Ramberg-Osgood本构模型中硬化指数n的各种计算方法,提出采用快速退火算法(fast simulated annealing,FSA)求解n的方法并且编制计算程序,从而获得,n的置信区间,所得结果比传统两点法更稳定.在此基础上得到了国产6082-T6高强铝合金材料本构模型,与试验实测应力-应变曲线的比较表明,提出的本构模型具有很好的预测性,可为铝合金结构的工程应用提供参考.%Material constitutive relationship is one of the basic problems for analysis and application of aluminum alloy structures.This paper systematically studied the material constitutive model of the domestic high-strength new grade aluminum alloy 6082-T6.Tensile testing was conducted for a total of 90 coupons that were cut from three cross-section shapes of the aluminum alloy 6082-T6.The measurement and statistical analysis of mechanical parameters were carried out,including non-proportional extension strengths,tensile strength,elastic modulus,breaking elongation,Poisson' s ratio,etc.Various methods for calculating hardening exponent n in the Ramberg-Osgood law were discussed.The fast simulated annealing (FSA) method for calculating n was proposed and the FSA code was written to obtain the confidence interval of n.Comparison of results between FSA and other methods indicates that the FSA can provide more stable results than the traditional two-point method.On this basis,the material constitutive model of the 6082-T6 aluminum alloy was obtained.The comparison of this model and the tested stress-strain curve demonstrates that the constitutive model

  10. Micromechanics of Composite Materials Governed by Vector Constitutive Laws

    Science.gov (United States)

    Bednarcyk, Brett A.; Aboudi, Jacob; Arnold, Steven M.

    2017-01-01

    The high-fidelity generalized method of cells micromechanics theory has been extended for the prediction of the effective property tensor and the corresponding local field distributions for composites whose constituents are governed by vector constitutive laws. As shown, the shear analogy, which can predict effective transverse properties, is not valid in the general three-dimensional case. Consequently, a general derivation is presented that is applicable to both continuously and discontinuously reinforced composites with arbitrary vector constitutive laws and periodic microstructures. Results are given for thermal and electric problems, effective properties and local field distributions, ordered and random microstructures, as well as complex geometries including woven composites. Comparisons of the theory's predictions are made to test data, numerical analysis, and classical expressions from the literature. Further, classical methods cannot provide the local field distributions in the composite, and it is demonstrated that, as the percolation threshold is approached, their predictions are increasingly unreliable. XXXX It has been observed that the bonding between the fibers and matrix in composite materials can be imperfect. In the context of thermal conductivity, such imperfect interfaces have been investigated in micromechanical models by Dunn and Taya (1993), Duan and Karihaloo (2007), Nan et al. (1997) and Hashin (2001). The present HFGMC micromechanical method, derived for perfectly bonded composite materials governed by vector constitutive laws, can be easily generalized to include the effects of weak bonding between the constituents. Such generalizations, in the context of the mechanical micromechanics problem, involve introduction of a traction-separation law at the fiber/matrix interface and have been presented by Aboudi (1987), Bednarcyk and Arnold (2002), Bednarcyk et al. (2004) and Aboudi et al. (2013) and will be addressed in the future.

  11. Neuroendocrine gastric carcinoma expressing somatostatin: A highly malignant, rare tumor

    Institute of Scientific and Technical Information of China (English)

    Jaques Waisberg; Leandro Luongo de Matos; Ana Maria do Amaral Antonio Mader; Sérgio Pezzolo; Esmeralda Miristene Eher; Vera Luiza Capelozzi; Manlio Basilio Speranzini

    2006-01-01

    Poorly differentiated gastric neuroendocrine carcinomas,although rare, deserve particular attention, as they are aggressive and have an extremely poor prognosis. In this report we describe a gastric neuroendocrine carcinoma with rapidly fatal outcome. Immunohistological staining of the resected specimens revealed that the tumor was an endocrine carcinoma. The tumor disclosed intense immunoreactivity to pan-neuroendocrine markers and diffuse somatostatin immunoreactivity. There were no psammoma bodies and no demonstrable association with von Recklinghausen's neurofibromatosis. In the gastrointestinal tract, neuroendocrine tumors producing predominantly somatostatin have been described only in the duodenum. To the best of our knowledge, the present report is the second case report of a neuroendocrine gastric carcinoma expressing diffusely somatostatin as the only neuroendocrine regulatory peptide.

  12. A comparative analysis of constitutive promoters located in adeno-associated viral vectors.

    Directory of Open Access Journals (Sweden)

    Lkhagvasuren Damdindorj

    Full Text Available The properties of constitutive promoters within adeno-associated viral (AAV vectors have not yet been fully characterized. In this study, AAV vectors, in which enhanced GFP expression was directed by one of the six constitutive promoters (human β-actin, human elongation factor-1α, chicken β-actin combined with cytomegalovirus early enhancer, cytomegalovirus (CMV, simian virus 40, and herpes simplex virus thymidine kinase, were constructed and introduced into the HCT116, DLD-1, HT-1080, and MCF-10A cell lines. Quantification of GFP signals in infected cells demonstrated that the CMV promoter produced the highest GFP expression in the six promoters and maintained relatively high GFP expression for up to eight weeks after infection of HCT116, DLD-1, and HT-1080. Exogenous human CDKN2A gene expression was also introduced into DLD-1 and MCF-10A in a similar pattern by using AAV vectors bearing the human β-actin and the CMV promoters. The six constitutive promoters were subsequently placed upstream of the neomycin resistance gene within AAV vectors, and HCT116, DLD-1, and HT-1080 were infected with the resulting vectors. Of the six promoters, the CMV promoter produced the largest number of G418-resistant colonies in all three cell lines. Because AAV vectors have been frequently used as a platform to construct targeting vectors that permit gene editing in human cell lines, we lastly infected the three cell lines with AAV-based targeting vectors against the human PIGA gene in which one of the six promoters regulate the neomycin resistance gene. This assay revealed that the CMV promoter led to the lowest PIGA gene targeting efficiency in the investigated promoters. These results provide a clue to the identification of constitutive promoters suitable to express exogenous genes with AAV vectors, as well as those helpful to conduct efficient gene targeting using AAV-based targeting vectors in human cell lines.

  13. Parallel Gene Expression Differences between Low and High Latitude Populations of Drosophila melanogaster and D. simulans.

    Science.gov (United States)

    Zhao, Li; Wit, Janneke; Svetec, Nicolas; Begun, David J

    2015-05-01

    Gene expression variation within species is relatively common, however, the role of natural selection in the maintenance of this variation is poorly understood. Here we investigate low and high latitude populations of Drosophila melanogaster and its sister species, D. simulans, to determine whether the two species show similar patterns of population differentiation, consistent with a role for spatially varying selection in maintaining gene expression variation. We compared at two temperatures the whole male transcriptome of D. melanogaster and D. simulans sampled from Panama City (Panama) and Maine (USA). We observed a significant excess of genes exhibiting differential expression in both species, consistent with parallel adaptation to heterogeneous environments. Moreover, the majority of genes showing parallel expression differentiation showed the same direction of differential expression in the two species and the magnitudes of expression differences between high and low latitude populations were correlated across species, further bolstering the conclusion that parallelism for expression phenotypes results from spatially varying selection. However, the species also exhibited important differences in expression phenotypes. For example, the genomic extent of genotype × environment interaction was much more common in D. melanogaster. Highly differentiated SNPs between low and high latitudes were enriched in the 3' UTRs and CDS of the geographically differently expressed genes in both species, consistent with an important role for cis-acting variants in driving local adaptation for expression-related phenotypes.

  14. High-level expression of Proteinase K from Tritirachium album Limber in Pichia pastoris using multi-copy expression strains.

    Science.gov (United States)

    Yang, Hu; Zhai, Chao; Yu, Xianhong; Li, Zhezhe; Tang, Wei; Liu, Yunyun; Ma, Xiaojian; Zhong, Xing; Li, Guolong; Wu, Di; Ma, Lixin

    2016-06-01

    Proteinase K is widely used in scientific research and industries. This report was aimed to achieve high-level expression of proteinase K using Pichia pastoris GS115 as the host strain. The coding sequence of a variant of proteinase K that has higher activity than the wild type protein was chosen and optimized based on the codon usage preference of P. pastoris. The novel open reading frame was synthesized and a series of multi-copy expression vectors were constructed based on the pHBM905BDM plasmid, allowing for the tandem integration of multiple copies of the target gene into the genome of P. pastoris with a single recombination. These strains were used to study the correlation between the gene copy number and the expression level of proteinase K. The results of quantitative polymerase chain reaction (qPCR) indicated that the tandem expression cassettes were integrated into the host genome stably. Meanwhile, the results of qPCR and enzyme activity assays indicated that the mRNA and protein expression levels of the target gene increased as the gene copy number increased. Moreover, the effect of gene dosage on the expression level of the recombinant protein was more obvious using high-density fermentation. The maximum expression level and enzyme activity of proteinase K, which were obtained from the recombinant yeast strain bearing 5 copies of the target gene after an 84-h induction, were approximately 8.069 mg/mL and 108,295 U/mL, respectively. The recombinant proteinase was purified and characterized. The optimum pH and temperature for the activity of this protease were approximately pH 11 and 55 °C, respectively.

  15. 高强钢先进成型技术和本构模型研究现状与发展趋势%Advanced Forming Process for High Strength Steel and Research Status and Development Tendency of Constitutive Model

    Institute of Scientific and Technical Information of China (English)

    孙蓟泉; 李双娇; 尹衍军

    2014-01-01

    High strength steel, the best choice for application in the lightweight technology of automotive body is limited to be used due to the forming technique. The principle, characteristics and present research developments of the advanced forming processes such as hydroforming tech-nology, laser butt welding technology and hot stamping technology are discussed. And also the constitutive model of the advanced high strength steel is explained in order to provide the basis for improving the forming property of the steel.%高强钢是汽车车身轻量化的首选材料,但其成型问题一直是限制其推广应用的重要因素。对高强钢的先进成型技术,如液压成型、激光拼焊、热冲压成型等的原理、特点及最新研究进展进行了论述。同时,对先进高强钢的本构模型进行了阐述,以期对改进高强钢成型性能提供依据。

  16. NLRP3 Inflammasome Expression and Signaling in Human Diabetic Wounds and in High Glucose Induced Macrophages

    Directory of Open Access Journals (Sweden)

    Xiaotian Zhang

    2017-01-01

    Full Text Available Introduction. To investigate the contribution and mechanism of NLRP3 inflammasome expression in human wounds in diabetes mellitus and in high glucose induced macrophages. Methods. In the present study, we compared the expression of NLRP3 inflammasome in debridement wound tissue from diabetic and nondiabetic patients. We also examined whether high glucose induces NLRP3 inflammasome expression in cultures THP-1-derived macrophages and the influence on IL-1β expression. Results. The expressions of NLRP3, caspase1, and IL-1β, at both the mRNA and protein level, were significantly higher in wounds of diabetic patients compared with nondiabetic wounds (P<0.05. High glucose induced a significant increase in NLRP3 inflammasome and IL-1β expression in THP-1-derived macrophages. M1 macrophage surface marker with CCR7 was significantly upregulated after high glucose stimulation. SiRNA-mediated silencing of NLRP3 expression downregulates the expression of IL-1β. Conclusion. The higher expression of NLRP3, caspase1, and secretion of IL-1β, signaling, and activation might contribute to the hyperinflammation in the human diabetic wound and in high glucose induced macrophages. It may be a novel target to treat the DM patients with chronic wound.

  17. Prediction of highly expressed genes in microbes based on chromatin accessibility

    DEFF Research Database (Denmark)

    Willenbrock, Hanni; Ussery, David

    2007-01-01

    BACKGROUND: It is well known that gene expression is dependent on chromatin structure in eukaryotes and it is likely that chromatin can play a role in bacterial gene expression as well. Here, we use a nucleosomal position preference measure of anisotropic DNA flexibility to predict highly expressed...... and ribosomal RNA are encoded by DNA having significantly lower position preference values than other genes in fast-replicating microbes. CONCLUSION: This insight into DNA structure-dependent gene expression in microbes may be exploited for predicting the expression of non-translated genes such as non...

  18. 29 CFR 452.18 - Constitutional officers.

    Science.gov (United States)

    2010-07-01

    ... 29 Labor 2 2010-07-01 2010-07-01 false Constitutional officers. 452.18 Section 452.18 Labor... DISCLOSURE ACT OF 1959 Coverage of Election Provisions § 452.18 Constitutional officers. A constitutional... identified as such or provided for in the constitution or other organic law of the labor organization. 17...

  19. Venus Express bistatic radar: High-elevation anomalous reflectivity

    Science.gov (United States)

    Simpson, Richard A.; Tyler, G. Leonard; Häusler, Bernd; Mattei, Riccardo; Pätzold, Martin

    2009-06-01

    Magellan (MGN) bistatic radar observations in 1994 confirmed earlier Pioneer Venus reports of unusual Venus surface reflectivity and emissivity at elevations above 6054 km radius. They also revealed that the anomalous values of surface dielectric constant $\\varepsilon$ near Cleopatra Patera included a large imaginary component ($\\varepsilon$ ≈ -i 100) at 13 cm wavelength, consistent with a semiconducting surface material. The MGN observations were conducted using a linearly polarized wave, canted at 45° with respect to the plane of incidence and radiated by the MGN synthetic aperture radar antenna toward the specularly reflecting region of the mean planetary surface. In 2006 similar experiments were conducted using 13 cm circularly polarized transmissions from Venus Express (VEX). The VEX signal-to-noise ratio (SNR) was lower than that of MGN, but elevated ∣$\\varepsilon$∣ has been inferred broadly over Maxwell Montes. A quasi-specular echo was detected near Cleopatra but with insufficient SNR to address the question of conductivity. An early failure of the VEX 13 cm radio system precludes further measurements with VEX.

  20. A 4-gene expression score associated with high levels of Wilms Tumor-1 (WT1) expression is an adverse prognostic factor in acute myeloid leukaemia

    NARCIS (Netherlands)

    Niavarani, A. (Ahmadreza); Herold, T. (Tobias); Y. Reyal (Yasmin); M.C. Sauerland (Maria); T. Büchner (Thomas); W. Hiddemann (Wolfgang); S.K. Bohlander (Stefan); P.J.M. Valk (Peter); Bonnet, D. (Dominique)

    2016-01-01

    textabstractWilms Tumor-1 (WT1) expression level is implicated in the prognosis of acute myeloid leukaemia (AML). We hypothesized that a gene expression profile associated with WT1 expression levels might be a good surrogate marker. We identified high WT1 gene sets by comparing the gene expression p

  1. High-throughput cloning and expression in recalcitrant bacteria

    NARCIS (Netherlands)

    Geertsma, Eric R.; Poolman, Bert

    2007-01-01

    We developed a generic method for high-throughput cloning in bacteria that are less amenable to conventional DNA manipulations. The method involves ligation-independent cloning in an intermediary Escherichia coli vector, which is rapidly converted via vector-backbone exchange (VBEx) into an organism

  2. Constitutive Effects of Performance Indicators

    DEFF Research Database (Denmark)

    Dahler-Larsen, Peter

    2014-01-01

    The idea that performance indicators in public management have unintended consequences is almost as old as performance measurement itself. But, is ‘unintended consequences’ an appropriate and insightful idea? The very term rests on an identification of intentions and assumptions about validity...... that are demonstrably problematic. Based on a distinction between trivial and advanced measure fixation, an argument is made for constitutive effects that are based on less problematic assumptions. Through this conceptual move, the political dimension of performance indicators is appreciated. The conceptual dimensions...

  3. Yeast expression proteomics by high-resolution mass spectrometry

    DEFF Research Database (Denmark)

    Walther, Tobias C; Olsen, Jesper Velgaard; Mann, Matthias

    2010-01-01

    -translational controls contribute majorly to regulation of protein abundance, for example in heat shock stress response. The development of new sample preparation methods, high-resolution mass spectrometry and novel bioinfomatic tools close this gap and allow the global quantitation of the yeast proteome under different...... conditions. Here, we provide background information on proteomics by mass-spectrometry and describe the practice of a comprehensive yeast proteome analysis....

  4. A unified viscoplasticity constitutive model based on irreversible thermodynamics

    Institute of Scientific and Technical Information of China (English)

    LIU ChangChun; LV HeXiang; GUAN Ping

    2008-01-01

    A unified viscoplasticity constitutive model for metal materials is developed within the framework of irreversible thermodynamics, and an expression for the Helmholtz free energy function involving the parameters reflecting kinematic hardening and isotropic hardening is given. At the same time a non-associated flow potential function including the corresponding state variables is also given, from which the flow equation and the evolution equations of the internal state variables are derived. Thus, a general theoretical framework constructing a unified viscoplasticity con-stitutive model is given. Compared with the typical unified viscoplasticity constitu-tive models, the presented model evidently satisfies the irreversible thermody-namics laws. Moreover, this method not only provides a new theoretical foundation for further development of the unified viscoplasticity constitutive model, but also gives a new theoretical framework for the stress-strain analysis of more materials.

  5. A unified viscoplasticity constitutive model based on irreversible thermodynamics

    Institute of Scientific and Technical Information of China (English)

    2008-01-01

    A unified viscoplasticity constitutive model for metal materials is developed within the framework of irreversible thermodynamics, and an expression for the Helmholtz free energy function involving the parameters reflecting kinematic hardening and isotropic hardening is given. At the same time a non-associated flow potential function including the corresponding state variables is also given, from which the flow equation and the evolution equations of the internal state variables are derived. Thus, a general theoretical framework constructing a unified viscoplasticity constitutive model is given. Compared with the typical unified viscoplasticity constitutive models, the presented model evidently satisfies the irreversible thermodynamics laws. Moreover, this method not only provides a new theoretical foundation for further development of the unified viscoplasticity constitutive model, but also gives a new theoretical framework for the stress-strain analysis of more materials.

  6. Differential expression of apolipoprotein D in male reproductive system of rats by high-fat diet.

    Science.gov (United States)

    Lim, W; Bae, H; Song, G

    2016-11-01

    Apolipoprotein D, a 29-kDa secreted glycoprotein that belongs to the lipocalin superfamily, is widely expressed in various tissues and associated with lipid metabolism as a component of high-density lipoproteins. Although Apolipoprotein D binds to small hydrophobic ligands including cholesterol, little is known about effects of high-fat diet with cholesterol on expression of Apolipoprotein D in the male reproductive tract. Therefore, we investigated Apod expression in penises, prostate glands, and testes from rats fed a high-fat diet including a high amount of cholesterol. Our previous research indicated that a high-fat diet induces dyslipidemia leading to histological changes and dysfunction of male reproduction in rats. Consistent with these results, Apod mRNA expression was significantly (p high-fat diet as compared with normal diet. In addition, Apod mRNA and protein were detected predominantly in urethral epithelium and penile follicle from rats. Moreover, changes in expression of specific microRNAs (miR-229b-3p, miR-423-3p, and miR-490-3p) regulating Apod in the penises and prostate glands were negatively associated with Apod expression. Collectively, results of this study suggest that Apod is a novel regulatory gene in the male reproductive system, especially in penises of rats fed a high-cholesterol diet, and that expression of Apod is regulated at the posttranscriptional level by target microRNAs.

  7. Apolipoprotein E expression and behavioral toxicity of high charge, high energy (HZE) particle radiation

    Energy Technology Data Exchange (ETDEWEB)

    Higuchi, Yoshinori; Nelson, G.A.; Slater, J.M.; Pearlstein, R.D. [Loma Linda Univ., CA (United States). Medical Center; Vazquez, M. [Brookhaven National Lab., Upton, NY (United States); Laskowitz, D.T. [Duke Univ., Durham, NC (United States). Medical Center

    2002-12-01

    Apolipoprotein E (apoE) is a lipid binding protein that plays an important role in tissue repair following brain injury. In the present studies, we have investigated whether apoE affects the behavioral toxicity of high charge, high energy (HZE) particle radiation. Sixteen male apoE knockout (KO) mice and sixteen genetically matched wild-type (WT) C57BL mice were used in this experiment. Half of the KO and half of the WT animals were irradiated with 600 MeV/amu iron particles (2 Gy whole body). The effect of irradiation on motor coordination and stamina (Rotarod test), exploratory behavior (open field test), and spatial working and reference memory (Morris water maze) was assessed. Rotarod test: Performance was adversely affected by radiation exposure in both KO and WT groups at 30 d after irradiation. By 60 d after radiation, the radiation effect was lost in WT, but still apparent in irradiated KO mice. Open field test: Radiation reduced open field exploratory activity 14, 28, 56, 84, and 168 d after irradiation of KO mice, but had no effect on WT mice. Morris water maze: Radiation adversely affected spatial working memory in the KO mice, but had no discernible effect in the WT mice as assessed 180 d after irradiation. In contrast, irradiated WT mice showed marked impairment of spatial reference memory in comparison to non-irradiated mice, while no effect of radiation was observed in KO mice. These studies show that apoE expression influences the behavioral toxicity of HZE particle radiation and suggest that apoE plays a role in the repair/recovery from radiation injury of the central nervous system (CNS). ApoE deficiency may exacerbate the previously reported effects of HZE particle radiation in accelerating the brain aging process. (author)

  8. Essence and constitution in Zubiri

    Directory of Open Access Journals (Sweden)

    Alfonso Gómez Fernández

    2015-03-01

    Full Text Available The aim of this article is to expound Zubiri’s concept of essence and to begin a critical discussion and evaluation of his proposal. Before explaining what essence is, it is necessary to observe that, for our author, not all things have an essence: only «reality-things» but not «meaning-things» form part of the «essentiable domain». This distinction between «reality-things» and «meaning-things» supposes a new conception of nature. (Nature is not the opposite of the artificial, as the Greek term physis is.«Reality-things» form a constitutional and «closed» system of non-causal and non-«external» characteristics or traits. These «foundational characteristics» of constitution, then, are precisely the essence. The article discusses critically the assumptions of this doctrine (reality, ontological pluralism, foundation and its possible limits in the light of contemporary science. It can be a useful theory for the philosophy of language.

  9. A Comparative Study on Johnson Cook, Modified Zerilli-Armstrong and Arrhenius-Type Constitutive Models to Predict High-Temperature Flow Behavior of Ti-6Al-4V Alloy in α + β Phase

    Science.gov (United States)

    Cai, Jun; Wang, Kuaishe; Han, Yingying

    2016-03-01

    True stress and true strain values obtained from isothermal compression tests over a wide temperature range from 1,073 to 1,323 K and a strain rate range from 0.001 to 1 s-1 were employed to establish the constitutive equations based on Johnson Cook, modified Zerilli-Armstrong (ZA) and strain-compensated Arrhenius-type models, respectively, to predict the high-temperature flow behavior of Ti-6Al-4V alloy in α + β phase. Furthermore, a comparative study has been made on the capability of the three models to represent the elevated temperature flow behavior of Ti-6Al-4V alloy. Suitability of the three models was evaluated by comparing both the correlation coefficient R and the average absolute relative error (AARE). The results showed that the Johnson Cook model is inadequate to provide good description of flow behavior of Ti-6Al-4V alloy in α + β phase domain, while the predicted values of modified ZA model and the strain-compensated Arrhenius-type model could agree well with the experimental values except under some deformation conditions. Meanwhile, the modified ZA model could track the deformation behavior more accurately than other model throughout the entire temperature and strain rate range.

  10. High-level expression of the native barley alpha-amylase/subtilisin inhibitor in Pichia pastoris

    DEFF Research Database (Denmark)

    Micheelsen, Pernille Ollendorff; Ostergaard, Peter Rahbek; Lange, Lene

    2008-01-01

    An expression system for high-level expression of the native Hordeum vulgare alpha-amylase/subtilisin inhibitor (BASI) has been developed in Pichia pastoris, using the methanol inducible alcohol oxidase 1 (AOX1) promoter. To optimize expression, two codon-optimized coding regions have been design...... and characterized by Edman degradation, liquid chromatography mass spectrometry and insoluble blue starch assay, and was shown to possess the same characteristics as wild-type protein purified from barley grains....

  11. Contesting a World-Constitution? Anti-Systemic Movements and Constitutional Forms in Ireland, 1848-2008

    Directory of Open Access Journals (Sweden)

    Thomas Murray

    2016-03-01

    Full Text Available Recent accounts of constitutional development have emphasised commonalities among diverse constitutions in terms of the transnational migration of legal institutions and ideas. World-systems analysis gives critical expression to this emergent intellectual trajectory. Since the late 18th century, successive, international waves of constitution-making have tended to correspond with decisive turning points in the contested formation of the historical capitalist world-system. The present article attempts to think through the nature of this correspondence in the Irish context. Changes to the Irish constitution, I suggest, owed to certain local manifestations of anti-systemic movements within the historical capitalist world-system and to constitution-makers’ attempts to contain – militarily, politically and ideologically – these movements’ democratic and egalitarian ideals and practices. Various configurations of the balance of power in Irish society between ‘national’ (core-peripheral and ‘social’ (capital-labour/‘other’ forces crystallised in constitutional form. Thus far, conservative and nationalist constitutional projects have tended to either dominate or incorporate social democratic and radical ones, albeit a process continually contested at critical junctures by civil society and by the organised left, both old and new.

  12. 高温三轴应力下气煤蠕变特征及本构模型%Creep characteristics and constitutive model of gas coal mass under high temperature and triaxial stress

    Institute of Scientific and Technical Information of China (English)

    周长冰; 万志军; 张源; 刘渝; 张博

    2012-01-01

    Utilizing the 20 MN servo-controlled triaxial rock testing machine with high temperature and high pressure developed by China University of Mining and Technology,experimental study was carried out on the creep properties of large size gas coal specimens of 200 mm×400 mm under two temperature points of triaxial stress at 200 ℃ and 400 ℃.The research results indicate that the gas coal experiences the first and second phases of creep at 200 ℃,but without acceleration of creep;and phase that creep accelerates appears after a short time at 400 ℃.In the joint action of temperature and stress,the gas coal specimens show typical sticky-elastic-plastic deformation;because of the moment and subsequent deformation in the loading process,coal specimens are always accompanied by significant plastic deformation.By the analysis of the permeability and porosity of gas coal under different temperatures,it was initially judged that 300 ℃ is the critical temperature of the change of creep characteristics about gas coal.On the basis of high-temperature creep characteristics of gas coal,a new type of nonlinear dashpots was introduced,and a creep constitutive model under high temperature aimed at gas coal was built,which theoretical curves of axial creep could agree well with the experimental curves,which indicating that this constitutive model can preferably simulate the creep characteristics of the gas coal under high temperature.%采用中国矿业大学研制的"600℃20 MN伺服控制高温高压岩体三轴试验机",对尺寸为200 mm×400 mm的山东兴隆庄气煤进行了200℃和400℃两个温度点的三轴应力下的蠕变实验。实验表明:200℃时,气煤蠕变具有明显的第1,2阶段,但未出现加速蠕变阶段,而400℃时,气煤在很短时间内就进入加速蠕变阶段;在温度和应力共同作用下,气煤在加载瞬间及后续变形过程中,始终伴随着显著的塑性变形,煤体变形为典型的黏弹塑性变形。通过

  13. Towards constitutive equations for the deep Earth

    Science.gov (United States)

    Kennett, B. L. N.

    2017-09-01

    A new formulation of constitutive equations for states of high compression is introduced for isotropic media, exploiting a separation between hydrostatic and deviatoric components in strain energy. The strain energy is represented as functions of strain invariants, with one purely volumetric component and the other which vanishes for purely hydrostatic deformation. This approach preserves the form of familiar equations of state through the volumetric component, but allows the addition of volume and pressure dependence of the shear modulus from the deviatoric term. A suitable shear modulus representation to accompany a Keane equation of state is demonstrated.

  14. High Expression of Cry1Ac Protein in Cotton (Gossypium hirsutum by Combining Independent Transgenic Events that Target the Protein to Cytoplasm and Plastids.

    Directory of Open Access Journals (Sweden)

    Amarjeet Kumar Singh

    Full Text Available Transgenic cotton was developed using two constructs containing a truncated and codon-modified cry1Ac gene (1,848 bp, which was originally characterized from Bacillus thuringiensis subspecies kurstaki strain HD73 that encodes a toxin highly effective against many lepidopteran pests. In Construct I, the cry1Ac gene was cloned under FMVde, a strong constitutively expressing promoter, to express the encoded protein in the cytoplasm. In Construct II, the encoded protein was directed to the plastids using a transit peptide taken from the cotton rbcSIb gene. Genetic transformation experiments with Construct I resulted in a single copy insertion event in which the Cry1Ac protein expression level was 2-2.5 times greater than in the Bacillus thuringiensis cotton event Mon 531, which is currently used in varieties and hybrids grown extensively in India and elsewhere. Another high expression event was selected from transgenics developed with Construct II. The Cry protein expression resulting from this event was observed only in the green plant parts. No transgenic protein expression was observed in the non-green parts, including roots, seeds and non-green floral tissues. Thus, leucoplasts may lack the mechanism to allow entry of a protein tagged with the transit peptide from a protein that is only synthesized in tissues containing mature plastids. Combining the two events through sexual crossing led to near additive levels of the toxin at 4-5 times the level currently used in the field. The two high expression events and their combination will allow for effective resistance management against lepidopteran insect pests, particularly Helicoverpa armigera, using a high dosage strategy.

  15. Freedom of Expression Laws and the College Press: Lessons Learned from the High Schools.

    Science.gov (United States)

    Paxton, Mark

    This paper examines two recent attempts to enact state freedom of expression laws for public college and university students and discusses the prospects for such laws in the context of state scholastic freedom of expression laws covering high school journalists in six states. It examines the case of Kincaid v. Gibson, which decided that…

  16. NLRP3 Inflammasome Expression and Signaling in Human Diabetic Wounds and in High Glucose Induced Macrophages

    Science.gov (United States)

    Zhang, Xiaotian; Dai, Jiezhi; Li, Li

    2017-01-01

    Introduction. To investigate the contribution and mechanism of NLRP3 inflammasome expression in human wounds in diabetes mellitus and in high glucose induced macrophages. Methods. In the present study, we compared the expression of NLRP3 inflammasome in debridement wound tissue from diabetic and nondiabetic patients. We also examined whether high glucose induces NLRP3 inflammasome expression in cultures THP-1-derived macrophages and the influence on IL-1β expression. Results. The expressions of NLRP3, caspase1, and IL-1β, at both the mRNA and protein level, were significantly higher in wounds of diabetic patients compared with nondiabetic wounds (P CCR7 was significantly upregulated after high glucose stimulation. SiRNA-mediated silencing of NLRP3 expression downregulates the expression of IL-1β. Conclusion. The higher expression of NLRP3, caspase1, and secretion of IL-1β, signaling, and activation might contribute to the hyperinflammation in the human diabetic wound and in high glucose induced macrophages. It may be a novel target to treat the DM patients with chronic wound. PMID:28164132

  17. Highly expressed genes are associated with inverse antisense transcription in mouse

    Indian Academy of Sciences (India)

    Andras Györffy; Pawel Surowiak; Zsolt Tulassay; Balazs Györffy

    2007-08-01

    There is a growing evidence, that antisense transcription might have a key role in a range of human diseases. Although predefined sense–antisense pairs were extensively studied, the antisense expression of the known sense genes is rarely investigated. We retrieved and correlated the expression of sense and antisense sequences of 1182 mouse transcripts to assess the prevalence and to find the characteristic pattern of antisense transcription. We contrasted three Affymetrix MGU74A version 1 mouse genome chips to six MGU74A version 2 chips. For these 1182 transcripts, the version 1 chips contain the antisense sequences of the transcripts presented on the version 2 chips. The original data was taken from the GEO database (GDS431 and GDS432). As the Affymetrix data are semiquantitative, the relative expression levels of antisense partners were analysed. We detected antisense transcription, although the average antisense expression is shifted towards smaller expression values (MGU74A version 1, 516; version 2, 1688). An inverse direct correlation between sense and antisense expression values could be observed at high expression values. At a very high relative expression—above 40,000—the Pearson correlation coefficient is getting closer to −1. Transcripts with high inverse expression ratio may be correlated to the investigated gene (major histocompatibility complex class II trans activator). The ratio of sense to antisense transcripts varied among different chromosomes; on chromosomes 14 and 1 the level of antisense expression was higher than that of sense. We conclude that antisense transcription is a common phenomenon in the mouse genome. The hypothesis of regulatory role of antisense transcripts is supported by the inverse antisense gene expression of highly expressed genes.

  18. SAMSN1 is highly expressed and associated with a poor survival in glioblastoma multiforme.

    Directory of Open Access Journals (Sweden)

    Yong Yan

    Full Text Available OBJECTIVES: To study the expression pattern and prognostic significance of SAMSN1 in glioma. METHODS: Affymetrix and Arrystar gene microarray data in the setting of glioma was analyzed to preliminarily study the expression pattern of SAMSN1 in glioma tissues, and Hieratical clustering of gene microarray data was performed to filter out genes that have prognostic value in malignant glioma. Survival analysis by Kaplan-Meier estimates stratified by SAMSN1 expression was then made based on the data of more than 500 GBM cases provided by The Cancer Genome Atlas (TCGA project. At last, we detected the expression of SAMSN1 in large numbers of glioma and normal brain tissue samples using Tissue Microarray (TMA. Survival analysis by Kaplan-Meier estimates in each grade of glioma was stratified by SAMSN1 expression. Multivariate survival analysis was made by Cox proportional hazards regression models in corresponding groups of glioma. RESULTS: With the expression data of SAMSN1 and 68 other genes, high-grade glioma could be classified into two groups with clearly different prognoses. Gene and large sample tissue microarrays showed high expression of SAMSN1 in glioma particularly in GBM. Survival analysis based on the TCGA GBM data matrix and TMA multi-grade glioma dataset found that SAMSN1 expression was closely related to the prognosis of GBM, either PFS or OS (P<0.05. Multivariate survival analysis with Cox proportional hazards regression models confirmed that high expression of SAMSN1 was a strong risk factor for PFS and OS of GBM patients. CONCLUSION: SAMSN1 is over-expressed in glioma as compared with that found in normal brains, especially in GBM. High expression of SAMSN1 is a significant risk factor for the progression free and overall survival of GBM.

  19. Growth hormone (GH)-independent dimerization of GH receptor by a leucine zipper results in constitutive activation

    DEFF Research Database (Denmark)

    Behncken, S N; Billestrup, Nils; Brown, R;

    2000-01-01

    Growth hormone initiates signaling by inducing homodimerization of two GH receptors. Here, we have sought to determine whether constitutively active receptor can be created in the absence of the extracellular domain by substituting it with high affinity leucine zippers to create dimers of the gro......Growth hormone initiates signaling by inducing homodimerization of two GH receptors. Here, we have sought to determine whether constitutively active receptor can be created in the absence of the extracellular domain by substituting it with high affinity leucine zippers to create dimers...... proliferation after interleukin 3 withdrawal at a rate equal to maximally stimulated wild type GHR-expressing cells. Activation of STAT 5b was also observed in Fos-Jun-GHR-expressing cells at a level equal to that in chronically GH-treated GHR-expressing cells. Thus, forced dimerization of the transmembrane...... and cytoplasmic domains of the GHR in the absence of the extracellular domain can lead to the constitutive activation of known GH signaling end points, supporting the view that proximity of Janus kinase 2 (JAK2) kinases is the essential element in signaling. Such constitutively active GH receptors may have...

  20. Constitutive model for overconsolidated clays

    Institute of Scientific and Technical Information of China (English)

    2008-01-01

    Based on the relationships between the Hvorslev envelope,the current yield sur-face and the reference yield surface,a new constitutive model for overconsolidated clays is proposed. It adopts the unified hardening parameter,to which the potential failure stress ratio and the characteristic state stress ratio are introduced. The model can describe many characteristics of overconsolidated clays,including stress-strain relationships,strain hardening and softening,stress dilatancy,and stress path dependency. Compared with the Cam-clay model,the model only re-quires one additional soil parameter which is the slope of the Hvorslev envelope. Comparisons with data from triaxial drained compression tests for Fujinomori clay show that the proposed model can rationally describe overconsolidated properties. In addition,the model is also used to predict the stress-strain relationship in the isotropic consolidation condition and the stress paths in the undrained triaxial compression tests.

  1. Connexin 30.3 is expressed in the kidney but not regulated by dietary salt or high blood pressure.

    Science.gov (United States)

    Hanner, Fiona; Schnichels, Marc; Zheng-Fischhöfer, Qingyi; Yang, Li E; Toma, Ildikó; Willecke, Klaus; McDonough, Alicia A; Peti-Peterdi, János

    2008-05-01

    Several isoforms of connexin (Cx) proteins have been identified in a variety of tissues where they play a role in intercellular communication, either as the components of gap junctions or as large, nonselective pores known as hemichannels. This investigation seeks to identify the localization and regulation of Cx30.3 in mouse, rat, and rabbit kidney using a Cx30.3(+/lacZ) transgenic approach and immunofluorescence. Cx30.3 was detected in all three species and predominantly in the renal medulla. Both the nuclear lacZ staining indicative of Cx30.3 expression and indirect immunohistochemistry provided the same results. Cx30.3 immunolabeling was mainly punctate in the mouse, typical for gap junctions. In contrast, it showed continuous apical plasma membrane localization in certain tubule segments in the rat and rabbit kidney, suggesting that it may also function as hemichannels. In the cortex, Cx30.3 was localized in the intercalated cells of the cortical collecting duct, because the immunoreactive cells did not label for AQP2, a marker for principal cells. In the medulla, dense Cx30.3 staining was confined to the ascending thin limbs of the loop of Henle, because the immunoreactive cells did not label for AQP1, a marker of the descending thin limbs. Immunoblotting studies indicated that Cx30.3 expression was unchanged in response to either high or low salt intake or in spontaneously hypertensive rats. Cx30.3 appears to be constitutively expressed in certain renal tubular segments and cells and its role in overall kidney function remains to be investigated.

  2. Evaluation of P-glycoprotein (Pgp) expression in human osteosarcoma by high-throughput tissue microarray.

    Science.gov (United States)

    Gao, Yan; Liao, Yunfei; Shen, Jacson K; Feng, Yong; Choy, Edwin; Cote, Gregory; Harmon, David; Mankin, Henry J; Hornicek, Francis J; Duan, Zhenfeng

    2016-09-01

    Survival of osteosarcoma patients is currently limited by the development of metastases and multidrug resistance (MDR). A well-established cause of MDR involves overexpression of P-glycoprotein (Pgp) in tumor cells. However, some discrepancies still exist as to the clinical significance of Pgp in osteosarcoma. We sought to elucidate further whether the Pgp expression correlated with clinical behavior in a series of patients with osteosarcoma via high-throughput tissue microarray (TMA) analysis. Immunohistochemical analysis of Pgp expression in a TMA of 114 specimens with a retrospective review of 70 osteosarcoma patients admitted to the Massachusetts General Hospital (MGH) was performed. High Pgp expression was correlated with metastasis development and poor response to pre-operative chemotherapy in osteosarcoma patients. Eighteen of the fifty-seven patients initially admitted with primary osteosarcoma showed high Pgp expression. Among these 18 patients with high Pgp expression, 13 of 18 (72%) patients eventually developed metastases. There was no significant clinical relevance between Pgp expression and osteosarcoma survival. These results support that high expression of Pgp is important, but cannot be assigned as, an individual predictor in the development of human osteosarcoma. © 2016 Orthopaedic Research Society. Published by Wiley Periodicals, Inc. J Orthop Res 34:1606-1612, 2016. © 2016 Orthopaedic Research Society. Published by Wiley Periodicals, Inc.

  3. 75 FR 57835 - Constitution Day and Citizenship Day, Constitution Week, 2010

    Science.gov (United States)

    2010-09-22

    ... Proclamation 8562--Constitution Day and Citizenship Day, Constitution Week, 2010 Proclamation 8563--National... President ] Proclamation 8562 of September 16, 2010 Constitution Day and Citizenship Day, Constitution Week... Convention in Philadelphia established a Constitution for the United States of America, signing the...

  4. The Obligations on Government and Society in our Constitutional State to Respect and Support Independent Constitutional Structures

    Directory of Open Access Journals (Sweden)

    LWH Ackermann

    2000-05-01

    must be independent. Its members cannot be elected, because that would imply that the Court owed allegiance or were accountable to the political majority or other elector in question. On the other hand, it is seen as undemocratic for a body that is not elected to be in a position to overrule the expressed will of the political representatives of the majority. This paradox exists in respect of all our courts and makes the method of appointing judicial officers particularly important in order to ensure at the same time, and as far as this is practically possible, both their independence and their legitimacy. The judiciary is however not an arm of the state that has been exempted from all checks and balances. The checks and balances on the judiciary are not the same as in the case of the legislature and the executive. In the case of the latter the checks and balances are principally through the Constitution, as enforced by the courts, and through the political process. In the case of the courts these checks and balances cannot be through the political process, for this would undermine the independence of the judiciary. One of the reciprocal obligations that a constitutional democracy imposes on all its subjects, is to support the independent constitutional institutions, as constitutional institutions, not only vocally at the level of intellectual abstraction, but by actively working to establish the habits of consitutionalism in all societal structures and societal interaction

  5. High Immunoglobulin A Levels Mediate the Association Between High Anger Expression and Low Somatic Symptoms in Intimate Partner Violence Perpetrators.

    Science.gov (United States)

    Romero-Martínez, A; Lila, M; Vitoria-Estruch, S; Moya-Albiol, L

    2016-02-01

    It has been hypothesized that anger expression may be associated with increased salivary immunoglobulin A (sIgA) levels, which is associated with decreased somatic symptoms, and therefore anger expression may be associated with reduced somatic symptoms in intimate partner violence (IPV) perpetrators. This study tested the potential mediating effect of sIgA levels on the relationship between anger expression and respiratory and gastrointestinal symptoms in IPV perpetrators and non-violent controls. The sample consisted of IPV perpetrators (n = 19) and controls (n = 21). Saliva samples were collected for assessing sIgA levels. The State-Trait Anger Expression Inventory-2 was used to assess anger expression and the Revised version of the Somatic Symptoms Scale developed by Sandín and Chorot to measure somatic symptoms. High anger expression was associated with low levels of respiratory and gastrointestinal symptoms in IPV perpetrators mediated through high sIgA levels but the same was not true for non-violent controls. This finding supports the hypothesis that for IPV perpetrators, anger expression may be physiologically and psychologically rewarding. Future research examining other immunological parameters is needed to further test this hypothesis. Such effort may illuminate why some IPV perpetrators continue to use violence against their partners.

  6. Ezrin is highly expressed in early thymocytes, but dispensable for T cell development in mice.

    Directory of Open Access Journals (Sweden)

    Meredith H Shaffer

    Full Text Available BACKGROUND: Ezrin/radixin/moesin (ERM proteins are highly homologous proteins that function to link cargo molecules to the actin cytoskeleton. Ezrin and moesin are both expressed in mature lymphocytes, where they play overlapping roles in cell signaling and polarity, but their role in lymphoid development has not been explored. METHODOLOGY/PRINCIPAL FINDINGS: We characterized ERM protein expression in lymphoid tissues and analyzed the requirement for ezrin expression in lymphoid development. In wildtype mice, we found that most cells in the spleen and thymus express both ezrin and moesin, but little radixin. ERM protein expression in the thymus was differentially regulated, such that ezrin expression was highest in immature thymocytes and diminished during T cell development. In contrast, moesin expression was low in early thymocytes and upregulated during T cell development. Mice bearing a germline deletion of ezrin exhibited profound defects in the size and cellularity of the spleen and thymus, abnormal thymic architecture, diminished hematopoiesis, and increased proportions of granulocytic precursors. Further analysis using fetal liver chimeras and thymic transplants showed that ezrin expression is dispensable in hematopoietic and stromal lineages, and that most of the defects in lymphoid development in ezrin(-/- mice likely arise as a consequence of nutritional stress. CONCLUSIONS/SIGNIFICANCE: We conclude that despite high expression in lymphoid precursor cells, ezrin is dispensable for lymphoid development, most likely due to redundancy with moesin.

  7. Evidence against the energetic cost hypothesis for the short introns in highly expressed genes

    Directory of Open Access Journals (Sweden)

    Niu Deng-Ke

    2008-05-01

    Full Text Available Abstract Background In animals, the moss Physcomitrella patens and the pollen of Arabidopsis thaliana, highly expressed genes have shorter introns than weakly expressed genes. A popular explanation for this is selection for transcription efficiency, which includes two sub-hypotheses: to minimize the energetic cost or to minimize the time cost. Results In an individual human, different organs may differ up to hundreds of times in cell number (for example, a liver versus a hypothalamus. Considered at the individual level, a gene specifically expressed in a large organ is actually transcribed tens or hundreds of times more than a gene with a similar expression level (a measure of mRNA abundance per cell specifically expressed in a small organ. According to the energetic cost hypothesis, the former should have shorter introns than the latter. However, in humans and mice we have not found significant differences in intron length between large-tissue/organ-specific genes and small-tissue/organ-specific genes with similar expression levels. Qualitative estimation shows that the deleterious effect (that is, the energetic burden of long introns in highly expressed genes is too negligible to be efficiently selected against in mammals. Conclusion The short introns in highly expressed genes should not be attributed to energy constraint. We evaluated evidence for the time cost hypothesis and other alternatives.

  8. High chondroitin sulfate proteoglycan 4 expression correlates with poor outcome in patients with breast cancer.

    Science.gov (United States)

    Hsu, Nicholas C; Nien, Pei-Yung; Yokoyama, Kazunari K; Chu, Pei-Yi; Hou, Ming-Feng

    2013-11-15

    Chondroitin sulfate proteoglycan 4 (CSPG4), a transmembrane proteoglycan originally identified in melanoma cells, has been reported to be expressed in breast cancer cells. This study was performed to examine the expression and significance of CSPG4 in a cohort of breast cancer patients. Immunohistochemical analysis of CSPG4 was performed on tissue microarrays constructed from tissue specimens from 240 breast cancer patients. CSPG4 staining was correlated with clinical and pathological characteristics, overall survival (OS), and disease recurrence. Contradicting to a previous report, our results showed that high CSPG4 expression was not related to triple-negative status of breast cancer patients. The Kaplan-Meier method showed that high CSPG4 expression was significantly associated with shorter time to recurrence (TTR). Patients with high CSPG4 expression had poorer OS and shorter TTR in a multivariate survival analysis after adjustment for stage, tumor grade, expression of estrogen receptor and progesterone receptor, and HER2 overexpression. This study showed that high CSPG4 expression correlates with disease recurrence and OS in breast cancers.

  9. The communicative constitution of representation and exclusion

    NARCIS (Netherlands)

    Wessel, van M.G.J.

    2014-01-01

    In the light of current debate on representation, specifically engaging with literature showing how representation is communicatively constituted, this paper empirically shows how exclusion also can be seen as communicatively constituted. The interpretive approach toward communication employed in th

  10. Constitutional Analysis of Educational Vouchers in Minnesota.

    Science.gov (United States)

    Bruno, Robert J.

    1989-01-01

    Voucher plans that involve payments of tax funds to private schools are suspect on both constitutional and public policy grounds. Analyzes the constitutional issues raised by a recent attempt at voucher legislation in Minnesota. (MLF)

  11. Native Communities and the Peruvian Constitutional Assembly

    Science.gov (United States)

    American Indian Journal, 1978

    1978-01-01

    A loosely knit coalition of over 25 native groups, the Peruvian Amazon Peoples has prepared a statement directed at the Peruvian Constitutional Assembly for purposes of Native input into the preparation of a revised national constitution. (JC)

  12. The communicative constitution of representation and exclusion

    NARCIS (Netherlands)

    Wessel, van M.G.J.

    2014-01-01

    In the light of current debate on representation, specifically engaging with literature showing how representation is communicatively constituted, this paper empirically shows how exclusion also can be seen as communicatively constituted. The interpretive approach toward communication employed in th

  13. 118 CONSTITUTIONALISM AND GOOD GOVERNANCE IN NIGERIA

    African Journals Online (AJOL)

    Fr. Ikenga

    Concentration of governmental powers in the hands of one individual is the very .... resources.39 The idea of good or bad governance is an ethical one. ... Despite the fact that on the face value, the constitution proclaims constitutionalism, the actions.

  14. Constitutional design and political agency problems: the case of Colombia

    Directory of Open Access Journals (Sweden)

    Paola Alarcón López

    2014-06-01

    Full Text Available This article presents and explores, from a law and economics’ perspective, the Colombian Constitutional structures that create potential for corruption by undermining the checks and balances, the accountability system over politicians and in particular, by altering the computation of the agent once is facing the decision to act according to principal interest or behave in a corrupt way. A case study from a Colombian high court is presented in order to illustrate the interaction between the identified constitutional provisions to generate a scope for corruption. Finally, possible constitutional amendments are suggested in order to overcome these system failures.

  15. Constitutional design and political agency problems: the case of Colombia

    Directory of Open Access Journals (Sweden)

    Paola Alarcón López

    2014-07-01

    Full Text Available This article presents and explores, from a law and economics’ perspective, the Colombian Constitutional structures that create potential for corruption by undermining the checks and balances, the accountability system over politicians and in particular, by altering the computation of the agent once is facing the decision to act according to principal interest or behave in a corrupt way. A case study from a Colombian high court is presented in order to illustrate the interaction between the identified constitutional provisions to generate a scope for corruption. Finally, possible constitutional amendments are suggested in order to overcome these system failures.

  16. Constitutive theories on viscoelastoplasticity and damage of frozen soil

    Institute of Scientific and Technical Information of China (English)

    何平; 程国栋; 朱元林

    1999-01-01

    The constitutive theory on the viscoelastoplasticity and damage of frozen soil is based on the continuous mechanics and thermodynamics. The basic principles of the theory, dissipation potential function and damage model are presented. The constitutive theory explains the mechanical properties of frozen soils under complicated stresses, especially under high confining pressures which make frozen soil harden and soften. The agreement between the calculated results by the constitutive theory and the experimental results of triaxial creep of frozen soil is seen to be very good.

  17. Constitutional Principles of State Control in the Republic of Armenia

    Directory of Open Access Journals (Sweden)

    Gabriel K. Balayan

    2015-12-01

    Full Text Available In the present article some of the provision of the RA Constitution’s basic principles of state control, which, in my opinion, should be guided by regulatory authorities during the inspection are discussed. These principles are considered by the author as the basis of the constitutionality of state control. Some of the control principles are divided by the author into positive and negative. Positive defines the principles that provide the freedom of action of man and citizen. Negative same principles are for government agencies, since they restrict their activities. A separate analysis is subject to the presidency on the basis of the principle of separation and balance of powers. In conclusion, author concludes that, based on the interpretation of the constitutional principles discussed, the term "law" applies only to natural and legal persons on the other hand can not be used as an analogue of the terms "authority" and "authority." According to the author, the fundamental constitutional principles of state control are expressed formally in a single system as defined in the first chapter of the Constitution of the Republic of Armenia. Compliance and reform of constitutional principles provide the synergy and the constitutionality of the process of governance.

  18. Global gene expression profiling reveals genes expressed differentially in cattle with high and low residual feed intake.

    Science.gov (United States)

    Chen, Y; Gondro, C; Quinn, K; Herd, R M; Parnell, P F; Vanselow, B

    2011-10-01

    Feed efficiency is an economically important trait in beef production. It can be measured as residual feed intake. This is the difference between actual feed intake recorded over a test period and the expected feed intake of an animal based on its size and growth rate. DNA-based marker-assisted selection would help beef breeders to accelerate genetic improvement for feed efficiency by reducing the generation interval and would obviate the high cost of measuring residual feed intake. Although numbers of quantitative trait loci and candidate genes have been identified with the advance of molecular genetics, our understanding of the physiological mechanisms and the nature of genes underlying residual feed intake is limited. The aim of the study was to use global gene expression profiling by microarray to identify genes that are differentially expressed in cattle, using lines genetically selected for low and high residual feed intake, and to uncover candidate genes for residual feed intake. A long-oligo microarray with 24 000 probes was used to profile the liver transcriptome of 44 cattle selected for high or low residual feed intake. One hundred and sixty-one unique genes were identified as being differentially expressed between animals with high and low residual feed intake. These genes were involved in seven gene networks affecting cellular growth and proliferation, cellular assembly and organization, cell signalling, drug metabolism, protein synthesis, lipid metabolism, and carbohydrate metabolism. Analysis of functional data using a transcriptional approach allows a better understanding of the underlying biological processes involved in residual feed intake and also allows the identification of candidate genes for marker-assisted selection. © 2011 The Authors, Animal Genetics © 2011 Stichting International Foundation for Animal Genetics.

  19. A family of E. coli expression vectors for laboratory scale and high throughput soluble protein production

    Directory of Open Access Journals (Sweden)

    Bottomley Stephen P

    2006-03-01

    Full Text Available Abstract Background In the past few years, both automated and manual high-throughput protein expression and purification has become an accessible means to rapidly screen and produce soluble proteins for structural and functional studies. However, many of the commercial vectors encoding different solubility tags require different cloning and purification steps for each vector, considerably slowing down expression screening. We have developed a set of E. coli expression vectors with different solubility tags that allow for parallel cloning from a single PCR product and can be purified using the same protocol. Results The set of E. coli expression vectors, encode for either a hexa-histidine tag or the three most commonly used solubility tags (GST, MBP, NusA and all with an N-terminal hexa-histidine sequence. The result is two-fold: the His-tag facilitates purification by immobilised metal affinity chromatography, whilst the fusion domains act primarily as solubility aids during expression, in addition to providing an optional purification step. We have also incorporated a TEV recognition sequence following the solubility tag domain, which allows for highly specific cleavage (using TEV protease of the fusion protein to yield native protein. These vectors are also designed for ligation-independent cloning and they possess a high-level expressing T7 promoter, which is suitable for auto-induction. To validate our vector system, we have cloned four different genes and also one gene into all four vectors and used small-scale expression and purification techniques. We demonstrate that the vectors are capable of high levels of expression and that efficient screening of new proteins can be readily achieved at the laboratory level. Conclusion The result is a set of four rationally designed vectors, which can be used for streamlined cloning, expression and purification of target proteins in the laboratory and have the potential for being adaptable to a high

  20. Methods of Writing Constitutional Formulas

    Directory of Open Access Journals (Sweden)

    Raos, N.

    2012-09-01

    Full Text Available Chemical formulas, as well as any linguistic entity, have to fulfill two basic requirements – expressiveness and economy, i.e. they have to express the maximal meaning with minimal means. Besides, chemical formula, being a scientific notation, has not to convey vague and scientifically unapproved meanings. This article presents the development of various kinds of chemical formulas and discusses their meaning in the historical context. Special attention is paid to line notation, developed for computers (WLN, SMILES, InChI etc.. We also discuss Seymour B. Elk's "biparametric nomenclature", based on the concept of 3-simplex, which was claimed to be universally applicable to all classes of compounds.

  1. The influence of bovine milk high or low in isoflavones on hepatic gene expression in mice

    DEFF Research Database (Denmark)

    Skaanild, Mette Tingleff; Nielsen, Tina Skau

    2012-01-01

    in hepatic gene expression after dietary intake of milk high and low in isoflavones. In addition to pelleted feed female NMRI mice were offered water, water added either 17β-estradiol, equol, Tween 80, and milk high and low in isoflavone content for a week. Gene expression was analyzed using an array q......Isoflavones have generated much attention due to their potential positive effects in various diseases. Phytoestrogens especially equol can be found in bovine milk, as feed ration for dairy cows is comprised of plants containing phytoestrogens. The aim of this study was to analyze the changes......PCR kit. It was revealed that Tween 80 and 17β-estradiol upregulated both phase I and phase II genes to the same extent whereas equol alone, high and low isoflavone milk did not alter the expression of phase I genes but decreased the expression of phase II genes. This study shows that dietary isoflavones...

  2. Theoretical Issues of the Constitutional Regulation Mechanism

    Science.gov (United States)

    Zhussupova, Guldaray B.; Zhailyaubayev, Rassul T.; Ukin, Symbat K.; Shunayeva, Sylu M.; Nurmagambetov, Rachit G.

    2016-01-01

    The purpose of this research is to define the concept of "constitutional regulation mechanism." The definition of the concept of "constitutional regulation mechanism" will give jurists and legislators a theoretical framework for developing legal sciences, such as the constitutional law and the theory of state and law. The…

  3. State Constitutional Law: Teaching and Scholarship.

    Science.gov (United States)

    Williams, Robert F.

    1991-01-01

    State constitutional law is an emerging area for legal education, partly because of state supreme court decisions relying on state rather than federal constitutional law. Studying state constitutional law highlights similarities and diversity of legal and governmental systems. Interest in establishment of curricula and materials in state law is…

  4. Antithetical Ethics: Kenneth Burke and the Constitution.

    Science.gov (United States)

    Anderson, Virginia

    1995-01-01

    Shows how the textuality of the United States Constitution, the most venerable of classic democratic icons, might be exploited to nurture postmodern ethics. Shows how Kenneth Burke's reading of the Constitution accords with and augments the postmodern theories of J.-F. Lyotard and S. Jarratt. Discusses a postmodern Constitution and the…

  5. The Constitution of the Republic of Estonia

    Index Scriptorium Estoniae

    2009-01-01

    Sisaldab ka: The Constitution of the Republic of Estonia Amendment Act. The Constitution of the Republic of Estonia Implementation Act. Act to Amend the Constitution of the Republic of Estonia for Election of Local Government Councils for Term of Four Years

  6. The Constitution of the Republic of Estonia

    Index Scriptorium Estoniae

    2005-01-01

    Raamat sisaldab ka: The Constitution of the Republic of Estonia amendment act ; The Constitution of the Republic of Estonia implementation act ; Act to amend the Constitution of the Republic of Estonia for election of local government councils for term of four years

  7. The Constitution of the Republic of Estonia

    Index Scriptorium Estoniae

    2009-01-01

    Sisaldab ka: The Constitution of the Republic of Estonia Amendment Act. The Constitution of the Republic of Estonia Implementation Act. Act to Amend the Constitution of the Republic of Estonia for Election of Local Government Councils for Term of Four Years

  8. The Constitution of the Republic of Estonia

    Index Scriptorium Estoniae

    2005-01-01

    Raamat sisaldab ka: The Constitution of the Republic of Estonia amendment act ; The Constitution of the Republic of Estonia implementation act ; Act to amend the Constitution of the Republic of Estonia for election of local government councils for term of four years

  9. 7 CFR 718.201 - Farm constitution.

    Science.gov (United States)

    2010-01-01

    ... 7 Agriculture 7 2010-01-01 2010-01-01 false Farm constitution. 718.201 Section 718.201 Agriculture... Reconstitution of Farms, Allotments, Quotas, and Bases § 718.201 Farm constitution. (a) In order to implement... this section. The constitution and identification of land as a farm for the first time and...

  10. Selection for the compactness of highly expressed genes in Gallus gallus

    Directory of Open Access Journals (Sweden)

    Zhou Ming

    2010-05-01

    (n = 1105, and compared the first intron length and the average intron length between highly expressed genes (top 5% expressed genes and weakly expressed genes (bottom 5% expressed genes. We found that the first intron length and the average intron length in highly expressed genes are not different from that in weakly expressed genes. We also made a comparison between ubiquitously expressed genes and narrowly expressed somatic genes with similar expression levels. Our data demonstrated that ubiquitously expressed genes are less compact than narrowly expressed genes with the similar expression levels. Obviously, these observations can not be explained by mutational bias hypotheses either. We also found that the significant trend between genes' compactness and expression level could not be affected by local mutational biases. We argued that the selection of economy model is most likely one to explain the relationship between gene expression and gene characteristics in chicken genome. Conclusion Natural selection appears to favor the compactness of highly expressed genes in chicken genome. This observation can be explained by the selection of economy model. Reviewers This article was reviewed by Dr. Gavin Huttley, Dr. Liran Carmel (nominated by Dr. Eugene V. Koonin and Dr. Araxi Urrutia (nominated by Dr. Laurence D. Hurst.

  11. High expression of testes-specific protease 50 is associated with poor prognosis in colorectal carcinoma.

    Directory of Open Access Journals (Sweden)

    Lei Zheng

    Full Text Available BACKGROUND: Testes-specific protease 50 (TSP50 is normally expressed in testes and abnormally expressed in breast cancer, but whether TSP50 is expressed in colorectal carcinoma (CRC and its clinical significance is unclear. We aimed to detect TSP50 expression in CRC, correlate it with clinicopathological factors, and assess its potential diagnostic and prognostic value. METHODOLOGY/PRINCIPAL FINDINGS: TSP50 mRNAs and proteins were detected in 7 CRC cell lines and 8 CRC specimens via RT-PCR and Western blot analysis. Immunohistochemical analysis of TSP50, p53 and carcinoembryonic antigen (CEA with tissue microarrays composed of 95 CRCs, 20 colorectal adenomas and 20 normal colorectal tissues were carried out and correlated with clinicopathological characteristics and disease-specific survival for CRC patients. There was no significant correlation between the expression levels of TSP50 and p53 (P = 0.751 or CEA (P = 0.663. Abundant expression of TSP50 protein was found in CRCs (68.4% while it was poorly expressed in colorectal adenomas and normal tissues (P<0.0001. Thus, CRCs can be distinguished from them with high specificity (92.5% and positive predictive value (PPV, 95.6%. The survival of CRC patients with high TSP50 expression was significantly shorter than that of the patients with low TSP50 expression (P = 0.010, specifically in patients who had early-stage tumors (stage I and II; P = 0.004. Multivariate Cox regression analysis indicated that high TSP50 expression was a statistically significant independent risk factor (hazard ratio  = 2.205, 95% CI = 1.214-4.004, P = 0.009. CONCLUSION: Our data demonstrate that TSP50 is a potential effective indicator of poor survival for CRC patients, especially for those with early-stage tumors.

  12. High expression level of soluble SARS spike protein mediated by adenovirus in HEK293 cells

    Institute of Scientific and Technical Information of China (English)

    Fei Zhong; Zhen-Yu Zhong; Shuang Liang; Xiu-Jin Li

    2006-01-01

    AIM: To develop a highly efficacious method for preparation of soluble SARS S-protein using adenovirus vector to meet the requirement for S-protein investigation.METHODS: The human adenovirus vector was used to express the soluble S-protein (corresponding to 1~1190 amino acids) fused with Myc/His tag using codon-optimized gene construct in HEK239 cells. The recombinant adenovirus bearing S-protein gene was generated by ligation method. The expressed S-protein with Myc/His tag was purified from culture medium with Ni-NTA agarose beads followed by dialysis. The S-protein was detected by Western blot and its biologic activity was analyzed by binding to Vero cells.RESULTS: Under the conditions of infection dose (MOI of 50) and expression time (48 h), the high-level expression of S-protein was obtained. The expression level was determined to be approximately 75 μg/106cells after purification. Purified soluble S-protein was readily detected by Western blot with anti-Myc antibody and showed the ability to bind to surface of Vero cells,demonstrating that the soluble S-protein could remain the biologic activity in the native molecule.CONCLUSION: The high-level expression of S-protein in HEK293 cells mediated by adenovirus can be achieved under the optimized expression conditions. The proteins possess the biologic activity, which lays a foundation for further investigation of S-protein biological function.

  13. High KRT8 expression promotes tumor progression and metastasis of gastric cancer.

    Science.gov (United States)

    Fang, Jian; Wang, Hao; Liu, Yun; Ding, Fangfang; Ni, Ying; Shao, Shihe

    2017-02-01

    Keratin8 (KRT8) is the major component of the intermediate filament cytoskeleton and predominantly expressed in simple epithelial tissues. Aberrant expression of KRT8 is associated with multiple tumor progression and metastasis. However, the role of KRT8 in gastric cancer (GC) remains unclear. In this study, KRT8 expression was investigated and it was found to be upregulated along with human GC progression and metastasis at both mRNA and protein levels in human gastric cancer tissues. In addition, KRT8 overexpression enhanced the proliferation and migration of human gastric cancer cells, whereas the knock-down of KRT8 by siRNA only inhibited migration of human gastric cancer cells. Integrinβ1-FAK-induced epithelial-mesenchymal-transition (EMT) only existed in the high KRT8 cells. Furthermore, KRT8 overexpression led to increase in p-smad2/3 levels and TGFβ dependent signaling events. KRT8 expression in GC was related to tumor clinical stage and worse survival. Kaplan-Meier analysis proved that KRT8 was associated with overall survival of patients with GC that patients with high KRT8 expression tend to have unfavorable outcome. Moreover, Cox's proportional hazards analysis showed that high KRT8 expression was a prognostic marker of poor outcome. These results provided that KRT8 expression may therefore be a biomarker or potential therapeutic target to identify patients with worse survival.

  14. High CD133 Expression Is Associated with Worse Prognosis in Patients with Glioblastoma.

    Science.gov (United States)

    Zhang, Wei; Chen, Huanran; Lv, Shengqing; Yang, Hui

    2016-05-01

    The CD133 antigen has been identified as a putative stem cell marker in gliomas. However, the prognostic significance of CD133 expression in glioblastoma patients remained controversial. A meta-analysis of published data was performed to comprehensively assess the prognostic role of CD133 expression in glioblastoma patients. Publications assessing the prognostic significance of CD133 expression in glioblastoma patients were identified in PubMed, Embase, and Web of Science up to November 2014. The pooled hazard ratio (HR) with 95% confidence interval (95% CI) was calculated using meta-analysis to evaluate the prognostic significance of CD133 expression in glioblastoma. Ten studies with a total of 715 glioblastoma patients were included into the meta-analysis. Overall, high CD133 expression was associated with poorer overall survival in patients with glioblastoma (HR = 1.96, 95% CI 1.46-2.64, P glioblastoma (HR = 2.03, 95% CI 1.43-2.88, P glioblastoma. Meta-analysis of studies with adjusted estimates further showed that high CD133 expression was an independent prognostic factor of glioblastoma. High CD133 expression is associated with worse prognosis in patients with glioblastoma. More prospective studies with well-design are needed to confirm this finding.

  15. High expression of p300 is linked to aggressive features and poor prognosis of Nasopharyngeal Carcinoma

    Directory of Open Access Journals (Sweden)

    Liao Zhi-Wei

    2012-05-01

    Full Text Available Abstract Background Increased expression of transcriptional coactivator p300 has been observed in a variety of human cancers. However, the expression status of p300 protein/mRNA in nasopharyngeal carcinoma (NPC tissues and its clinicopathologic/prognostic implication are poorly understood. Methods In our study, mRNA and protein expression levels of p300 was explored by reverse transcription-polymerase chain reaction (RT-PCR, Western blotting (WB and immunohistochemistry (IHC in nasopharyngeal mucosal and NPC tissues. The data were analyzed by receiver operating characteristic (ROC curve analysis, spearman’s rank correlation, Kaplan-Meier plots and Cox proportional hazards regression model. Results Up-regulated expression of p300 mRNA/p300 protein was detected in NPC tissues by RT-PCR and WB, when compared to nasopharyngeal mucosal tissues. Based on ROC curve analysis, the cutoff score for p300 high expression was defined when more than 35% of the tumor cells were positively stained. High expression of p300 was observed in 127/209 (60.7% of NPCs. In NPCs, high expression of p300 was positively associated with later T classification, later N classification, distant metastasis and later clinical stage (P P = 0.002 and overall survival (P = 0.001 in NPCs. More importantly, p300 expression was evaluated as an independent prognostic factor for NPC in multivariate analysis (P = 0.036. Conclusions Our findings support that high expression of p300 protein might be important in conferring a more aggressive behavior, and is an independent molecular marker for shortened survival time of patients with NPC.

  16. Differential localization and high expression of SURVIVIN splice variants in human embryonic stem cells but not in differentiated cells implicate a role for SURVIVIN in pluripotency

    Directory of Open Access Journals (Sweden)

    Amber N. Mull

    2014-03-01

    Full Text Available The BIRC5 gene encodes the oncofetal protein SURVIVIN, as well as four additional splice variants (ΔEx3, 2B, 3B and 2α. SURVIVIN, an inhibitor of apoptosis, is also a chromosomal passenger protein (CPP. Previous results have demonstrated that SURVIVIN is expressed at high levels in embryonic stem cells and inhibition of SURVIVIN function results in apoptosis, however these studies have not investigated the other four splice variants. In this study, we demonstrate that all variants are expressed at significantly higher levels in human embryonic stem (hES cells than in differentiated cells. We examined the subcellular localization of the three most highly expressed variants. SURVIVIN displayed canonical CPP localization in mitotic cells and cytoplasmic localization in interphase cells. In contrast, SURVIVIN–ΔEx3 and SURVIVIN–2B did not localize as a CPP; SURVIVIN–ΔEx3 was found constitutively in the nucleus while SURVIVIN–2B was distributed along the chromosomes during mitosis and also to the mitotic spindle poles. We used inducible shRNA against SURVIVIN to inhibit expression in a titratable fashion. Using this system, we reduced the mRNA levels of these three variants to approx. 40%, resulting in a concomitant reduction of OCT4 and NANOG mRNA, suggesting a role for the SURVIVIN variants in pluripotency.

  17. Seizure-related 6,a brain-specific expression gene,is highly expressed in the human cerebellum

    Institute of Scientific and Technical Information of China (English)

    Jianming Jiang; Long Yu; Yangtai Guan; Zhiliang Yu; Xinghua Huang; Xiaosong Chen; Lisha Tang; Xianning Zhang

    2010-01-01

    Epilepsy is a complex,Mendelian disease,and most cases are sporadic.Genomic comparisons of tissue from identified monogenic epilepsies with multigenic and acquired syndromes could ultimately reveal crucial molecular neuropathology for an epileptic phenotype.In the present study,a novel gene,human seizure-related(hSEZ)-6,was isolated from a human brain cDNA library.hSEZ-6 comprises 17 exons and spans a region of at least 55.6 kb,which was localized to 17q12 by radiation hybridization,hSEZ-6 exhibits two isoform types,hSEZ-6A and hSEZ-6B,which encode996 and 995 amino acids,respectively.The two putative hSEZ-6 proteins contain similar motifs and share 82% and 84% identity with mouse SEZ-6A protein,whose expression level increased in mouse cerebral cortex-derived cells treated with a convulsant drug,pentylentetrazole.Northern blot analysis demonstrated that hSEZ-6 is expressed highly in the cerebellum and in nucleus of the extrapyramidal system,such as the caudate nucleus and putamen.Reverse transcription polymerase chain reaction revealed that hSEZ-6 is expressed in neurons rather than gliocytes,which suggests that hSEZ-6 is a seizure-related gene.

  18. Reduced Popdc3 expression correlates with high risk and poor survival in patients with gastric cancer

    Institute of Scientific and Technical Information of China (English)

    Deng Luo; Ming-Liang Lu; Gong-Fang Zhao; Hua Huang; Meng-Yao Zheng; Jiang Chang; Lin Lv

    2012-01-01

    AIM:To investigate the expression of Popeye domain containing 3 (Popdc3) and its correlation with clinicopathological features and prognosis of gastric cancer.METHODS:The method of immunohistochemistry was used to investigate the expression of Popdc3 in 306 cases of human gastric cancer and 84 noncancerous gastric tissues.Simultaneously,the relationship between Popdc3 expression and the survival of the patients was retrospectively analyzed.RESULTS:Popdc3 was detected in 72 (85.71%) of 84 human nontumor mucosa.High expression of Popdc3 protein was detected in 78 (25.49%) of 306 human gastric cancer cases,and low expression was detected in 228 (74.51%).Low expression of Popdc3 correlated with depth of invasion (P < 0.0001),regional lymph nodes (P < 0.0001) and distant metastasis (P =0.02),and tumor,nodes,metastasis (TNM) stages (P< 0.0001).On multivariate analysis,only the patient's gender,regional lymph node metastasis,distant metastasis,TNM stages,and the expression of Popdc3 were independent prognostic factors in patients with gastric cancer.The Kaplan-Meier plot showed that low Popdc3 expression had a much more significant effect on the survival of those patients with early-stage tumors (x2 =104.741,P < 0.0001),with a > 51.9% reduction in the three-year survival compared with high Popdc3 expression.In late stages,the difference was also significant (x2 =5.930,P =0.015),with a 32.6% reduction in the three-year survival.CONCLUSION:Reduced expression of Popdc3 may play a significant role in the carcinogenesis and progression of gastric cancer.Popdc3 may be an independent prognostic factor.

  19. Expression of NDRG2 is down-regulated in high-risk adenomas and colorectal carcinoma

    DEFF Research Database (Denmark)

    Lorentzen, Anders; Vogel, Lotte K.; Lewinsky, Rikke H;

    2007-01-01

    BACKGROUND: It has recently been shown that NDRG2 mRNA is down-regulated or undetectable in several human cancers and cancer cell-lines. Although the function of NDRG2 is unknown, high NDRG2 expression correlates with improved prognosis in high-grade gliomas. The aim of this study has been to exa...

  20. Endothelial lipase is highly expressed in macrophages in advanced human atherosclerotic lesions

    DEFF Research Database (Denmark)

    Bartels, Emil D; Nielsen, John E; Lindegaard, Marie Louise Skakkebæk

    2007-01-01

    RNA expression increased markedly when either type of monocytes was differentiated into macrophages. Upon further differentiation into foam cells EL mRNA decreased whereas protein levels remained high compared to monocytes. In conclusion, macrophages in advanced human atherosclerotic lesions display high levels...

  1. High ubiquitous mitochondrial creatine kinase expression in hepatocellular carcinoma denotes a poor prognosis with highly malignant potential.

    Science.gov (United States)

    Uranbileg, Baasanjav; Enooku, Kenichiro; Soroida, Yoko; Ohkawa, Ryunosuke; Kudo, Yotaro; Nakagawa, Hayato; Tateishi, Ryosuke; Yoshida, Haruhiko; Shinzawa, Seiko; Moriya, Kyoji; Ohtomo, Natsuko; Nishikawa, Takako; Inoue, Yukiko; Tomiya, Tomoaki; Kojima, Soichi; Matsuura, Tomokazu; Koike, Kazuhiko; Yatomi, Yutaka; Ikeda, Hitoshi

    2014-05-01

    We previously reported the increased serum mitochondrial creatine kinase (MtCK) activity in patients with hepatocellular carcinoma (HCC), mostly due to the increase in ubiquitous MtCK (uMtCK), and high uMtCK mRNA expression in HCC cell lines. We explored the mechanism(s) and the relevance of high uMtCK expression in HCC. In hepatitis C virus core gene transgenic mice, known to lose mitochondrial integrity in liver and subsequently develop HCC, uMtCK mRNA and protein levels were increased in HCC tissues but not in non-tumorous liver tissues. Transient overexpression of ankyrin repeat and suppressor of cytokine signaling box protein 9 (ASB9) reduced uMtCK protein levels in HCC cells, suggesting that increased uMtCK levels in HCC cells may be caused by increased gene expression and decreased protein degradation due to reduced ASB9 expression. The reduction of uMtCK expression by siRNA led to increased cell death, and reduced proliferation, migration and invasion in HCC cell lines. Then, consecutive 105 HCC patients, who underwent radiofrequency ablation with curative intent, were enrolled to analyze their prognosis. The patients with serum MtCK activity >19.4 U/L prior to the treatment had significantly shorter survival time than those with serum MtCK activity ≤ 19.4 U/L, where higher serum MtCK activity was retained as an independent risk for HCC-related death on multivariate analysis. In conclusion, high uMtCK expression in HCC may be caused by hepatocarcinogenesis per se but not by loss of mitochondrial integrity, of which ASB9 could be a negative regulator, and associated with highly malignant potential to suggest a poor prognosis.

  2. A highly efficient pipeline for protein expression in Leishmania tarentolae using infrared fluorescence protein as marker

    Directory of Open Access Journals (Sweden)

    Mueller-Roeber Bernd

    2010-05-01

    . The facile in-cell and in-gel detection tools built on IFP make Leishmania amenable for high-throughput expression of proteins from plant and animal sources.

  3. CONSTITUTIVE RELATION OF UNSATURATED SOIL BY USE OF THE MIXTURE THEORY ( Ⅱ )-LINEAR CONSTITUTIVE EQUATIONS AND FIELD EQUATIONS

    Institute of Scientific and Technical Information of China (English)

    黄义; 张引科

    2003-01-01

    The linear constitutive equations and field equations of unsaturated soils were obtained through linearizing the nonlinear equations given in the first part of this work. The linear equations were expressed in the forms similar to Biot' s equations for saturated porous media. The Darcy's laws of unsaturated soil were proved. It is shown that Biot's equations of saturated porous media are the simplification of the theory. All these illustrate that constructing constitutive relation of unsaturated soil on the base of mixture theory is rational.

  4. Loi constitutive chimioplastique pour le beton expose aux hautes temperatures

    Science.gov (United States)

    Hammoud, Rabah

    degradation of exothermic origin. This experimental program puts emphasis on the fragile nature of the preheated concrete and demonstrates the non-applicability of two failure criteria often used in engineering calculation. An alternative is proposed and well-tested. Indeed, exposing the concrete to high temperature results in irreversible loss of stiffness as well as a loss of decohesion strength. These losses are, typically, expressed through semi-empirical relationships of the mechanical properties with temperature. Unfortunately, these relationships are inadequate because the direct impact of this degradation, on the macroscopic scale, can result in a dependency relationship between the elastic properties and the hydrates mass. Therefore, unlike traditional methods using conventional elasto-plastic models and adjusting certain parameters with local temperature, the proposed constitutive law that incorporates a function of dehydration similar to the softening index in chemo-plastics gives good results. An Etse and Willam similar criterion is used and modified for the occasion. Hardening and softening mechanisms are then needed to expand and contract the loading surface for defining the strength of the concrete on a wide range of dehydration processes. The direction and magnitude of a permanent deformation, core of the inelastic domain, are defined through the development of non-associated chemoplastic potential and new curve of ductility. The influence of hydrostatic pressure (dilatancy) and dehydration on the concrete behavior are taken into account in our model. The model is implemented in the Matlab(c) code. Strains and stresses generated in the concrete are now accurately predicted. To illustrate the capabilities of the developed model to predict the complex behavior of concrete exposed to high temperature, simulations are performed through numerical loading paths scenarios. The model is able to accurately reproduce all the experimental data.

  5. Establishment of Stable High Expression Cell Line with Green Fluorescent Protein and Resistance Genes

    Institute of Scientific and Technical Information of China (English)

    ZHANG Shengtao; LIU Wenli; HE Peigen; GONG Feili; YANG Dongliang

    2006-01-01

    In order to establish stable high expression cell lines, the eukaryotic expression vector pIRES2EGFP and recombinant plasmid pIRES2EGFP-TIM-3 were transfected into mammalian cells CHO by Lipofectamine. The transfected cells were cultivated under selective growth medium including G418 and green fluorescent protein (GFP) positive cells were sorted by FACS. Simultaneously, growing transfectants were selected only by G418 in the medium. The GFP expression in stably transfected cells was detected by FACS. Under selective growth conditions with G418, the percentage of GFP positive cells was reduced rapidly and GFP induction was low. In contrast, the percentages of GFP positive cells were increased gradually after FACS. By 3 rounds of GFP selection, the stable high expression cell lines were established. Furthermore, using FACS analysis GFP and the target protein TIM-3 co-expression in the stable transfectants cultured in nonselective medium was detected. Theses results demonstrated that the stably transfected cell lines that express high titer of recombinant protein can be simply and fleetly obtained by using GFP and selective growth medium.

  6. Stable Plastid Transformation for High-Level Recombinant Protein Expression: Promises and Challenges

    Directory of Open Access Journals (Sweden)

    Meili Gao

    2012-01-01

    Full Text Available Plants are a promising expression system for the production of recombinant proteins. However, low protein productivity remains a major obstacle that limits extensive commercialization of whole plant and plant cell bioproduction platform. Plastid genetic engineering offers several advantages, including high levels of transgenic expression, transgenic containment via maternal inheritance, and multigene expression in a single transformation event. In recent years, the development of optimized expression strategies has given a huge boost to the exploitation of plastids in molecular farming. The driving forces behind the high expression level of plastid bioreactors include codon optimization, promoters and UTRs, genotypic modifications, endogenous enhancer and regulatory elements, posttranslational modification, and proteolysis. Exciting progress of the high expression level has been made with the plastid-based production of two particularly important classes of pharmaceuticals: vaccine antigens, therapeutic proteins, and antibiotics and enzymes. Approaches to overcome and solve the associated challenges of this culture system that include low transformation frequencies, the formation of inclusion bodies, and purification of recombinant proteins will also be discussed.

  7. New inverse method for identification of constitutive parameters

    Institute of Scientific and Technical Information of China (English)

    2006-01-01

    A new inverse method by coupling iSIGHT and ABAQUS was proposed to determine the constitutive parameters of Al2O3sf/LY12 composite deforming at elevated temperature. It combined the merits of the finite element simulation and optimization technique. The direct model was simulated with finite element code ABAQUS. The inverse problem associated with the identification of the constitutive parameters was expressed as a least square optimization problem. The direct simulation and the parameters optimization were implemented in iSIGHT integrated environment. The aim was to match the output of the direct simulation with the experiment data.The capability of the proposed inverse method was demonstrated through the identification of constitutive parameters of Al2O3sf/LY12composite. The proposed new inverse method is also applicable to other parameters identification which is hardly determined through experiments or direct analytical method.

  8. 脲基酰胺酶基因在黄酒酵母中的整合型表达%Constitutive expression of DUR1 ,2 gene in Chinese rice wine yeast

    Institute of Scientific and Technical Information of China (English)

    朱旭亚; 陆健; 谢广发

    2012-01-01

    Ethyl carbamate(EC) is a potential carcinogen for human, which can be found in many fermented alcoholic beverages.The main precursor for EC is urea produced by yeast.The DUR1,2 gene which encoded urea amidolyase was cloned into plasmid pYX212 between the TPI promoter and terminator sites to form the DUR! ,2 expression cassette.The cassette was then integrated into the genome of the Chinese rice wine yeast 85#.Thus a urea-degrading Chinese rice wine yeast 85 was obtained.According to the laboratory scale rice wine brewing tests,urea was reduced to 8.34mg/L by yeast 85 69.9% less than the parental strain, while the ethyl carbamate level was 40.5% less than that of the parental strain after a period of storage.Meanwhile,85 was substantially equivalent to its parental strain in terms of fermentation ability, ethanol ,total acid and amino nitrogen.%氨基甲酸乙酯是一种人类的潜在致癌物,在许多发酵酒中均有存在,主要来源于发酵过程中酵母代谢产生的尿素。以pYX212为载体,将脲基酰胺酶基因DURl,2克隆到TPI强启动子和终止子之间的位点,再通过同源重组的方式将受强启动子调控的目的基因整合到黄酒酵母的基因组中,最终获得一株低产尿素的胞内脲基酰胺酶基因组成型高表达的黄酒酵母85在实验室规模的黄酒酿造实验中,85产尿素量为8.34mg/L,比出发菌株降低了69.9%,贮存一段时间后的酒液中氨基甲酸乙酯含量比出发菌株降低了40.5%,而发酵性能、酒精度、总酸及氨基态氮与出发菌株无显著差异,

  9. 78 FR 39721 - Constitution Pipeline Company, LLC; Notice of Application

    Science.gov (United States)

    2013-07-02

    ... Energy Regulatory Commission Constitution Pipeline Company, LLC; Notice of Application Take notice that on June 13, 2013, Constitution Pipeline Company, LLC (Constitution), having its principal place of...\\ Constitution further requests that the Commission grant Constitution a blanket certificate...

  10. State Constitutionalism: Completing the Interdisciplinary Study of Constitutional Law and Political Theory.

    Science.gov (United States)

    Williams, Robert F.

    1993-01-01

    Argues that a complete and accurate understanding of constitutional history and constitutional law requires the study of state constitutions. Maintains that state constitutions contain a coherent political theory that is, in important respects, at variance with the concept of federalism. (CFR)

  11. 77 FR 57981 - Constitution Day and Citizenship Day, Constitution Week, 2012

    Science.gov (United States)

    2012-09-18

    ... September 18, 2012 Part VI The President Proclamation 8862--Constitution Day and Citizenship Day, Constitution Week, 2012 #0; #0; #0; Presidential Documents #0; #0; #0;#0;Federal Register / Vol. 77 , No. 181... ] Proclamation 8862 of September 13, 2012 Constitution Day and Citizenship Day, Constitution Week, 2012 By...

  12. 3 CFR 8418 - Proclamation 8418 of September 16, 2009. Constitution Day and Citizenship Day, Constitution Week...

    Science.gov (United States)

    2010-01-01

    .... Constitution Day and Citizenship Day, Constitution Week, 2009 8418 Proclamation 8418 Presidential Documents Proclamations Proclamation 8418 of September 16, 2009 Proc. 8418 Constitution Day and Citizenship Day, Constitution Week, 2009By the President of the United States of America A Proclamation The United...

  13. 78 FR 57777 - Constitution Day and Citizenship Day, Constitution Week, 2013

    Science.gov (United States)

    2013-09-19

    ... September 19, 2013 Part IV The President Proclamation 9019--Constitution Day and Citizenship Day, Constitution Week, 2013 Proclamation 9020--Honoring the Victims of the Tragedy at the Washington Navy Yard #0... September 16, 2013 Constitution Day and Citizenship Day, Constitution Week, 2013 By the President of...

  14. The constitution of cometary nuclei

    Science.gov (United States)

    Whipple, F. L.

    1977-01-01

    The nongravitational term in the expression for the total force acting on a comet is calculated, and an upper limit is obtained for the product of the radial nongravitational term times the radius times the square root of the albedo. This condition is satisfied for ten periodic comets with q no greater than 1.5 AU, and the activity of these comets is consistent with control by H2O ice. Some of the comets must be spotty to account for their low albedo values. The effect of cosmic rays on comets, leading to frosting of their surface, is discussed.

  15. Monotreme lactation protein is highly expressed in monotreme milk and provides antimicrobial protection.

    Science.gov (United States)

    Enjapoori, Ashwantha Kumar; Grant, Tom R; Nicol, Stewart C; Lefèvre, Christophe M; Nicholas, Kevin R; Sharp, Julie A

    2014-09-22

    Monotremes (platypus and echidna) are the descendants of the oldest ancestor of all extant mammals distinguished from other mammals by mode of reproduction. Monotremes lay eggs following a short gestation period and after an even briefer incubation period, altricial hatchlings are nourished over a long lactation period with milk secreted by nipple-less mammary patches located on the female's abdomen. Milk is the sole source of nutrition and immune protection for the developing young until weaning. Using transcriptome and mass spectrometry analysis of milk cells and milk proteins, respectively, a novel Monotreme Lactation Protein (MLP) was identified as a major secreted protein in milk. We show that platypus and short-beaked echidna MLP genes show significant homology and are unique to monotremes. The MLP transcript was shown to be expressed in a variety of tissues; however, highest expression was observed in milk cells and was expressed constitutively from early to late lactation. Analysis of recombinant MLP showed that it is an N-linked glycosylated protein and biophysical studies predicted that MLP is an amphipathic, α-helical protein, a typical feature of antimicrobial proteins. Functional analysis revealed MLP antibacterial activity against both opportunistic pathogenic Staphylococcus aureus and commensal Enterococcus faecalis bacteria but showed no effect on Escherichia coli, Pseudomonas aeruginosa, Staphylococcus epidermidis, and Salmonella enterica. Our data suggest that MLP is an evolutionarily ancient component of milk-mediated innate immunity absent in other mammals. We propose that MLP evolved specifically in the monotreme lineage supporting the evolution of lactation in these species to provide bacterial protection, at a time when mammals lacked nipples.

  16. Monotreme Lactation Protein Is Highly Expressed in Monotreme Milk and Provides Antimicrobial Protection

    Science.gov (United States)

    Enjapoori, Ashwantha Kumar; Grant, Tom R.; Nicol, Stewart C.; Lefèvre, Christophe M.; Nicholas, Kevin R.; Sharp, Julie A.

    2014-01-01

    Monotremes (platypus and echidna) are the descendants of the oldest ancestor of all extant mammals distinguished from other mammals by mode of reproduction. Monotremes lay eggs following a short gestation period and after an even briefer incubation period, altricial hatchlings are nourished over a long lactation period with milk secreted by nipple-less mammary patches located on the female’s abdomen. Milk is the sole source of nutrition and immune protection for the developing young until weaning. Using transcriptome and mass spectrometry analysis of milk cells and milk proteins, respectively, a novel Monotreme Lactation Protein (MLP) was identified as a major secreted protein in milk. We show that platypus and short-beaked echidna MLP genes show significant homology and are unique to monotremes. The MLP transcript was shown to be expressed in a variety of tissues; however, highest expression was observed in milk cells and was expressed constitutively from early to late lactation. Analysis of recombinant MLP showed that it is an N-linked glycosylated protein and biophysical studies predicted that MLP is an amphipathic, α-helical protein, a typical feature of antimicrobial proteins. Functional analysis revealed MLP antibacterial activity against both opportunistic pathogenic Staphylococcus aureus and commensal Enterococcus faecalis bacteria but showed no effect on Escherichia coli, Pseudomonas aeruginosa, Staphylococcus epidermidis, and Salmonella enterica. Our data suggest that MLP is an evolutionarily ancient component of milk-mediated innate immunity absent in other mammals. We propose that MLP evolved specifically in the monotreme lineage supporting the evolution of lactation in these species to provide bacterial protection, at a time when mammals lacked nipples. PMID:25245409

  17. High-level gene expression in Aedes albopictus cells using a baculovirus Hr3 enhancer and IE1 transactivator

    Directory of Open Access Journals (Sweden)

    Gray Christine E

    2004-07-01

    Full Text Available Abstract Background Aedes aegypti is the key vector of both the Yellow Fever and Dengue Fever viruses throughout many parts of the world. Low and variable transgene expression levels due to position effect and position effect variegation are problematic to efforts to create transgenic laboratory strains refractory to these viruses. Transformation efficiencies are also less than optimal, likely due to failure to detect expression from all integrated transgenes and potentially due to limited expression of the transposase required for transgene integration. Results Expression plasmids utilizing three heterologous promoters and three heterologous enhancers, in all possible combinations, were tested. The Hr3/IE1 enhancer-transactivator in combination with each of the constitutive heterologous promoters tested increased reporter gene expression significantly in transiently transfected Aedes albopictus C7-10 cells. Conclusions The addition of the Hr3 enhancer to expression cassettes and concomitant expression of the IE1 transactivator gene product is a potential method for increasing the level of transgene expression in insect systems. This mechanism could also potentially be used to increase the level of transiently-expressed transposase in order to increase the number of integration events in transposon-mediated transformation experiments.

  18. High level of expression of recombinant human granulocyte-macrophage colony stimulating factor in transgenic rice cell suspension culture

    DEFF Research Database (Denmark)

    Shin, Yun-Ji; Hong, Shin-Young; Kwon, Tae-Ho;

    2003-01-01

    this problem, we sought an expression system in which heterologous gene expression could be induced at high levels. We selected a rice amylase expression system in which the promoter Ramy3D is induced to express recombinant protein by sucrose starvation. This induction system was found to give good yield...

  19. LXR-Mediated ABCA1 Expression and Function Are Modulated by High Glucose and PRMT2.

    Directory of Open Access Journals (Sweden)

    Maryem A Hussein

    Full Text Available High cholesterol and diabetes are major risk factors for atherosclerosis. Regression of atherosclerosis is mediated in part by the Liver X Receptor (LXR through the induction of genes involved in cholesterol transport and efflux. In the context of diabetes, regression of atherosclerosis is impaired. We proposed that changes in glucose levels modulate LXR-dependent gene expression. Using a mouse macrophage cell line (RAW 264.7 and primary bone marrow derived macrophages (BMDMs cultured in normal or diabetes relevant high glucose conditions we found that high glucose inhibits the LXR-dependent expression of ATP-binding cassette transporter A1 (ABCA1, but not ABCG1. To probe for this mechanism, we surveyed the expression of a host of chromatin-modifying enzymes and found that Protein Arginine Methyltransferase 2 (PRMT2 was reduced in high compared to normal glucose conditions. Importantly, ABCA1 expression and ABCA1-mediated cholesterol efflux were reduced in Prmt2-/- compared to wild type BMDMs. Monocytes from diabetic mice also showed decreased expression of Prmt2 compared to non-diabetic counterparts. Thus, PRMT2 represents a glucose-sensitive factor that plays a role in LXR-mediated ABCA1-dependent cholesterol efflux and lends insight to the presence of increased atherosclerosis in diabetic patients.

  20. Amino acid substitutions in the transcriptional regulator CbbR lead to constitutively active CbbR proteins that elevate expression of the cbb CO2 fixation operons in Ralstonia eutropha (Cupriavidus necator) and identify regions of CbbR necessary for gene activation.

    Science.gov (United States)

    Dangel, Andrew W; Tabita, F Robert

    2015-09-01

    CbbR is a LysR-type transcriptional regulator that activates expression of the operons containing (cbb) genes that encode the CO2 fixation pathway enzymes in Ralstonia eutropha (Cupriavidus necator) under autotrophic growth conditions. The cbb operons are stringently downregulated during chemoheterotrophic growth on organic acids such as malate. CbbR constitutive proteins (CbbR*s), typically with single amino acid substitutions, were selected and isolated that activate expression of the cbb operons under chemoheterotrophic growth conditions. A large set of CbbR*s from all major domains of the CbbR molecule were identified, except for the DNA-binding domain. The level of gene expression conferred for many of these CbbR*s under autotrophic growth was greater than that conferred by wild-type CbbR. Several of these CbbR*s increase transcription two- to threefold more than wild-type CbbR. One particular CbbR*, a truncated protein, was useful in identifying the regions of CbbR that are necessary for transcriptional activation and, by logical extension, necessary for interaction with RNA polymerase. The reductive assimilation of carbon via CO2 fixation is an important step in the cost-effective production of useful biological compounds. Enhancing CO2 fixation in Ralstonia eutropha through greater transcriptional activation of the cbb operons could prove advantageous, and the use of CbbR*s is one way to enhance product formation.

  1. High-level expression and purification of the human bradykinin B(2) receptor in a tetracycline-inducible stable HEK293S cell line.

    Science.gov (United States)

    Camponova, Paméla; Baud, Stéphanie; Mattras, Hélène; Duroux-Richard, Isabelle; Bonnafous, Jean-Claude; Marie, Jacky

    2007-10-01

    The B(2) bradykinin receptor belongs to the G-protein coupled receptor family. Development of new drugs for this important therapeutic target requires structural information on the receptor. The main goal of the present work was to overexpress the human B(2) receptor for future biophysical studies. Different tagged B(2) receptors were engineered and their properties were evaluated by transient expression in HEK293S cells. A B(2) receptor tagged with a hexahistidine at the N-terminus and a nonapeptide at the C-terminus was selected for high expression level and preserved ligand-binding characteristics. First, we generated a HEK293S stable cell line expressing the receptor constitutively at a level of 60pmol/mg of crude membrane protein. However, the decrease of expression level with cell passages led us to express the B(2) receptor in a HEK293S tetracycline-inducible stable cell line. Induction of expression of the B(2) receptor with tetracycline and sodium butyrate led to a level of 100pmol/mg of membrane protein, which is the highest level reported so far for this receptor. The expression level was stable with cell passages and the ligand-binding and signal transduction properties of the receptor were unaltered. The receptor was purified to near homogeneity by solubilization with n-dodecyl-beta-d-maltoside followed by a two-step purification procedure combining hydroxyapatite and immunoaffinity chromatography. Although the purified receptor is not functional, the purification of the B(2) receptor to near homogeneity from a stable cell line overexpressing this receptor pave the way for future structural studies of this receptor.

  2. Expression of Brassica napus L. γ-Glutamylcysteine Synthetase and Low-and High-Affinity Sulfate Transporters in Response to Excess Cadmium

    Institute of Scientific and Technical Information of China (English)

    Xin SUN; Xue-Mei SUN; Zhi-Min YANG; Shao-Qiong LI; Jin WANG; Song-Hua WANG

    2005-01-01

    In both the roots and leaves ofBrassica napus L. cv. Youyan No. 8 under treatment with 30 μmol/L Cd, massive production of non-protein thiols (NPT; mainly containing glutathione (GSH) and phytochelatins (PCs)) was induced, together with an increase in γ-glutamylcysteine synthetase (γ-ECS)mRNA transcripts. Because γ-ECS is the key enzyme catalyzing the first step in GSH biosynthesis, which, in turn, is converted to PCs, the Cd-induced increase in γ-ECS expression may be responsible for the observed increase in the production of NPT. Using a quantitative reverse transcription polymerase chain reaction (RT-PCR) approach, the expression of genes encoding a putative low-affinity sulfate transporter (LAST) and a putative high-affinity sulfate transporter (HAST) was determined at the transcriptional level. The RT-PCR analysis of relative transcript amounts indicates that the LAST gene in B. napus leaves showed a constitutive expression, which was hardly affected by Cd treatment. However, treatment with 30 μmol/L Cd for 2 or 3 d induced a marked increase in the expression of LAST in roots. Transcriptional expression of the HAST gene occurred in roots, but not in leaves. The expression of HAST only in the roots suggests that it has a specific function in sulfate uptake from soil and that the putative LAST may be responsible for the transport of sulfate from the roots to the shoots, as well as for the uptake of sulfate from soil. These results indicate that changes in transcriptional expression for sulfate transporters were required for the increased demand for sulfate during Cd stress.

  3. The Jurisdiction of The Constitutional Court In Controlling The Constitutionality of Criminal Judgments: Kosovo Case

    OpenAIRE

    BRAHA, Florentina Shala; BYTYQI, Vilard; VUNIQI, Dardan; Berisha, Fadil

    2016-01-01

    This paper will deal with individual requests that are submitted at the Constitutional Court in order to assess the constitutionality of criminal judicial decisions. The right to submit issues for assessing the constitutionality of criminal court decisions is a constitutionally guaranteed right. Many individuals who are dissatisfied with the decisions of the judicial instances use their right to oppose such a decision even at the Constitutional Court.Not all the cases submitted to the Court a...

  4. Expression and Effects of High-Mobility Group Box 1 in Cervical Cancer

    Directory of Open Access Journals (Sweden)

    Xiaoao Pang

    2014-05-01

    Full Text Available We investigated the significance of high- mobility group box1 (HMGB1 and T-cell-mediated immunity and prognostic value in cervical cancer. HMGB1, forkhead/winged helix transcription factor p3 (Foxp3, IL-2, and IL-10 protein expression was analyzed in 100 cervical tissue samples including cervical cancer, cervical intraepithelial neoplasia (CIN, and healthy control samples using immunohistochemistry. Serum squamous cell carcinoma antigen (SCC-Ag was immunoradiometrically measured in 32 serum samples from 37 cases of squamous cervical cancer. HMGB1 and SCC-Ag were then correlated to clinicopathological characteristics. HMGB1 expression tends to increase as cervical cancer progresses and it was found to be significantly correlated to FIGO stage and lymph node metastasis. These findings suggest that HMGB1 may be a useful prognostic indicator of cervical carcinoma. In addition, there were significant positive relationships between HMGB1 and FOXP3 or IL-10 expression (both p < 0.05. In contrast, HMGB1 and IL-2 expression was negatively correlated (p < 0.05. HMGB1 expression may activate Tregs or facilitate Th2 polarization to promote immune evasion of cervical cancer. Elevated HMGB1 protein in cervical carcinoma samples was associated with a high recurrence of HPV infection in univariate analysis (p < 0.05. HMGB1 expression and levels of SCC-Ag were directly correlated in SCC (p < 0.05. Thus, HMGB1 may be a useful biomarker for patient prognosis and cervical cancer prediction and treatment.

  5. Genomic Imprinting of IGF2 Is Maintained in Infantile Hemangioma despite its High Level of Expression

    Science.gov (United States)

    Yu, Ying; Wylie-Sears, Jill; Boscolo, Elisa; Mulliken, John B; Bischoff, Joyce

    2004-01-01

    Hemangioma, the most common tumor of infancy, is characterized by rapid growth and slow regression. Increased mRNA expression of insulin-like growth factor 2 (IGF2) has been detected in the proliferating phase by cDNA microarray analysis, but the underlying mechanism causing the increase remains unknown. Here, using quantitative real-time polymerase chain reaction (PCR) and immunohistochemistry, we show that IGF2 is highly expressed in both proliferating and involuting phase hemangioma, but is not detectable in other vascular lesions such as pyogenic granuloma, venous malformation, lymphatic malformation, or in normal infant skin. Loss of imprinting of the Igf2 gene has been associated with IGF2 overexpression in a variety of childhood tumors. To determine if loss of imprinting and consequent bi-allelic expression might contribute to the increased expression of IGF2, we examined the genomic imprinting status of Igf2 in 48 individual hemangiomas. We determined allele-specific Igf2 expression using reverse transcriptase–PCR combined with analysis of an Apa I–sensitive restriction fragment length polymorphism. Similar to heterozygous normal skin controls, all 15 informative hemangiomas showed uniform mono-allelic expression of Igf2. Therefore, loss of imprinting is not involved in the increased expression of IGF2 in infantile hemangioma. PMID:15706404

  6. Neoadjuvant multidrug chemotherapy including High-Dose Methotrexate modifies VEGF expression in Osteosarcoma: an immunohistochemical analysis

    Directory of Open Access Journals (Sweden)

    Rosa Michele A

    2010-02-01

    Full Text Available Abstract Background Angiogenesis plays a role in the progression of osteosarcoma, as well as in other mesenchymal tumors and carcinomas, and it is most commonly assessed by vascular endothelial growth factor (VEGF expression or tumor CD31-positive microvessel density (MVD. Tumor VEGF expression is predictive of poor prognosis, and chemotherapy can affect the selection of angiogenic pattern. The aim of the study was to investigate the clinical and prognostic significance of VEGF and CD31 in osteosarcoma, both at diagnosis and after neoadjuvant chemotherapy, in order to identify a potential role of chemotherapy in angiogenic phenotype. Methods A retrospective analysis was performed on 16 patients with high grade osteosarcoma. In each case archival pre-treatment biopsy tissue and post-chemotherapy tumor specimens were immunohistochemically stained against CD31 and VEGF, as markers of angiogenic proliferation both in newly diagnosed primary osteosarcoma and after multidrug chemotherapy including high-dose methotrexate (HDMTX. The correlation between clinicopathological parameters and the degree of tumor VEGF and CD31 expression was statistically assessed using the χ2 test verified with Yates' test for comparison of two groups. Significance was set at p Results Expression of VEGF was positive in 11 cases/16 of cases at diagnosis. Moreover, 8 cases/16 untreated osteosarcomas were CD31-negative, but the other 8 showed an high expression of CD31. VEGF expression in viable tumor cells after neoadjuvant chemotherapy was observed in all cases; in particular, there was an increased VEGF expression (post-chemotherapy VEGF - biopsy VEGF in 11 cases/16. CD31 expression increased in 11 cases/16 and decreased in 3 cases after chemotherapy. The data relating to the change in staining following chemotherapy appear statistically significant for VEGF expression (p p > 0,05. Conclusions Even if the study included few patients, these results confirm that VEGF and CD

  7. Analysis of DNA strand-specific differential expression with high density tiling microarrays

    Directory of Open Access Journals (Sweden)

    Antequera Francisco

    2010-03-01

    Full Text Available Abstract Background DNA microarray technology allows the analysis of genome structure and dynamics at genome-wide scale. Expression microarrays (EMA contain probes for annotated open reading frames (ORF and are widely used for the analysis of differential gene expression. By contrast, tiling microarrays (TMA have a much higher probe density and provide unbiased genome-wide coverage. The purpose of this study was to develop a protocol to exploit the high resolution of TMAs for quantitative measurement of DNA strand-specific differential expression of annotated and non-annotated transcripts. Results We extensively filtered probes present in Affymetrix Genechip Yeast Genome 2.0 expression and GeneChip S. pombe 1.0FR tiling microarrays to generate custom Chip Description Files (CDF in order to compare their efficiency. We experimentally tested the potential of our approach by measuring the differential expression of 4904 genes in the yeast Schizosaccharomyces pombe growing under conditions of oxidative stress. The results showed a Pearson correlation coefficient of 0.943 between both platforms, indicating that TMAs are as reliable as EMAs for quantitative expression analysis. A significant advantage of TMAs over EMAs is the possibility of detecting non-annotated transcripts generated only under specific physiological conditions. To take full advantage of this property, we have used a target-labelling protocol that preserves the original polarity of the transcripts and, therefore, allows the strand-specific differential expression of non-annotated transcripts to be determined. By using a segmentation algorithm prior to generating the corresponding custom CDFs, we identified and quantitatively measured the expression of 510 transcripts longer than 180 nucleotides and not overlapping previously annotated ORFs that were differentially expressed at least 2-fold under oxidative stress. Conclusions We show that the information derived from TMA

  8. Related high-risk factors of neonatal encephalopathy and its constitution analysis%新生儿脑病相关高危因素及构成分析

    Institute of Scientific and Technical Information of China (English)

    尚利宏; 王颍源; 李书津

    2014-01-01

    目的 分析新生儿脑病构成和相关危险因素,探讨降低新生儿脑损伤发生的措施.方法 选择3年内新生儿脑病患儿300例,对相关病因及高危因素进行分析.结果 入组的300例新生儿脑病中缺氧缺血性脑病(HIE)的比例仅为37.3%,导致新生儿脑病的原因还包括颅内出血、颅内感染、先天性遗传代谢性疾病、胆红素脑病等;单因素分析示胎儿宫内窘迫、胎盘异常、脐带异常、胎膜早破、围生期感染、败血症、难产等与新生儿缺氧缺血性脑病的发生有关,而产时的急性缺氧事件并非均导致缺氧缺血性脑病的发生.结论 新生儿脑病的构成包括缺氧缺血性脑病、颅内出血、颅内感染、先天性遗传性代谢病、胆红素脑病、染色体病、脑梗死、低血糖脑病、主要解剖异常等,而新生儿缺氧缺血性脑病与多种围生期的高危因素有关,因此不能把多数新生儿脑病的病因简单地归为产时的急性缺氧事件.%Objective To analyze the constitution and related risk factors of neonatal encephalopathy,and investigate the effective measures of reducing the occurrence of neonatal brain damage.Methods The clinical data of 300 cases of neonatal encephalopathy from January 2010 to December 2012 were retrospectively analyzed,the etioloaic factor and high risk factors of neonatal encephalopathy during the past three years were analyzed.Results Among the 300 cases of neonatal encephalopathy,hypoxic ischemic encephalopathy accounting for 37.3%,and the other risk factors of neonatal encephalopathy including intracranial hemorrhage,intracranial infection,congenital inherited metabolic diseases,bilirubin encephalopathy,and so on.Single factor analysis indicated that the following factors were associated with the development of neonatal hypoxic ischemic encephalopathy,including fetal distress in uterus,placenta,abnormality of umbilial cord,premature rupture of membranes

  9. Highly expressed genes in human high grade gliomas: immunohistochemical analysis of data from the Human Protein Atlas

    Directory of Open Access Journals (Sweden)

    Michael A. Meyer

    2014-06-01

    Full Text Available Gene expression within human glioblastomas were analyzed from data on 20,083 genes entered into the on-line Human Protein Atlas. In selecting genes that are strongly expressed within normal human brain tissue, 58 genes were identified from a search of the 20,083 entries that were rated as showing 90% or greater intensity of expression within normal brain tissues. Of these 58, a subset of 48 genes was identified that not only had expression data for human glioblastomas but also for the human glioblastoma cell line U-251. Four of these 48 selected genes were found to be strongly expressed within the cytoplasm when assessed by both histologic sampling of high grade glioma patient cases as well as U-251 glioblastoma cell line immunofluoresence analysis. These four human genes are: AGBL2 (ATP/GTP binding protein-like 2, BLOC1S6 (biogenesis of lysosomal organelles complex-1, subunit 6, MAP1A (microtubule-associated protein 1A and ZSWIM5 (zinc finger, SWIM-type containing 5, also known as KIAA1511. Further research is advocated to investigate the role of ZSWIM5 and AGBL2 in glioma cell biology.

  10. Testing of constitutive models in LAME.

    Energy Technology Data Exchange (ETDEWEB)

    Hammerand, Daniel Carl; Scherzinger, William Mark

    2007-09-01

    Constitutive models for computational solid mechanics codes are in LAME--the Library of Advanced Materials for Engineering. These models describe complex material behavior and are used in our finite deformation solid mechanics codes. To ensure the correct implementation of these models, regression tests have been created for constitutive models in LAME. A selection of these tests is documented here. Constitutive models are an important part of any solid mechanics code. If an analysis code is meant to provide accurate results, the constitutive models that describe the material behavior need to be implemented correctly. Ensuring the correct implementation of constitutive models is the goal of a testing procedure that is used with the Library of Advanced Materials for Engineering (LAME) (see [1] and [2]). A test suite for constitutive models can serve three purposes. First, the test problems provide the constitutive model developer a means to test the model implementation. This is an activity that is always done by any responsible constitutive model developer. Retaining the test problem in a repository where the problem can be run periodically is an excellent means of ensuring that the model continues to behave correctly. A second purpose of a test suite for constitutive models is that it gives application code developers confidence that the constitutive models work correctly. This is extremely important since any analyst that uses an application code for an engineering analysis will associate a constitutive model in LAME with the application code, not LAME. Therefore, ensuring the correct implementation of constitutive models is essential for application code teams. A third purpose of a constitutive model test suite is that it provides analysts with example problems that they can look at to understand the behavior of a specific model. Since the choice of a constitutive model, and the properties that are used in that model, have an enormous effect on the results of an

  11. Analysis on differential expressed genes of ovarian tissue between high- and low-yield laying hen.

    Science.gov (United States)

    Chen, Wei; Song, Ling-Jun; Zeng, Yong-Qing; Yang, Yun; Wang, Hui

    2013-01-01

    In order to elucidate molecular genetic mechanism of laying hen reproduction at the transcriptional level and the structure of significantly differential genes, the mRNA differential display and reverse northern dot-blot were used to detect the differential expression of genes in the ovary tissue of low-yield laying hens and high-yield laying hens in the present study. Sixteen 32-week-old CAU-pink laying hens divided into two groups were used and the laying performance was measured. The results showed that only the egg numbers were significantly different between the two groups; and from 15 primer pairs, a total of 336 bands were displayed of which 59 cDNA bands were found to be differentially expressed in both high-yield and low-yield laying hen. The sequence analysis indicated that the expression of such bands as H-AP5, H-P5, and H-P4 was significantly potentiated in high-yield laying hen using primer pairs AP5/HT11G, P5/HT11G and P4/HT11G and these transcripts had high homology (98%) to HoxDb, HoxCa, and HoxBa, respectively. The differentially expressed gene fragments may be relevant to the progression of the high-yield hens to the egg-laying stage. And further study is required to elucidate the molecular function to improve the productivity of laying hens.

  12. High expression of interleukin-22 and its receptor predicts poor prognosis in pancreatic ductal adenocarcinoma.

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    Wen, Zhang; Liao, Quan; Zhao, Jianguo; Hu, Ya; You, Lei; Lu, Zhaohui; Jia, Congwei; Wei, Yingxin; Zhao, Yupei

    2014-01-01

    The cytokine interleukin-22 (IL-22) and its receptor are present in the tumor microenvironment. Their function in pancreatic ductal adenocarcinoma (PDAC) remains largely unknown. The goal of the present study was to measure the expression of IL-22 and IL-22R in PDAC and assess their relationship with clinicopathological features and prognosis. The expression of IL-22 and IL-22R was evaluated by immunohistochemistry in PDAC tissues from 57 patients and by Western blotting in six tumors and adjacent nontumor tissues. A statistical analysis was conducted to assess the relationship between levels of expression, clinicopathological factors, and overall survival. In addition, the relationship between the expression of IL-22 and IL-22R and invasion was assessed by Western blotting and transwell assay with the PDAC cell lines PANC1 and BxPC3. Positive IL-22 staining was detected in PDAC tissues and adjacent nontumor tissues. Positive IL-22R staining was detected in PDAC cells. High expression of IL-22 and IL-22R correlated significantly with lymph node involvement. IL-22 increased the phosphorylation of signal transducer and activator of transcription3, the expression of matrix metalloproteinase 9, and the invasion in PANC1 and BxPC3 cells in vitro while silencing of IL-22R RNA caused opposite effects. Most importantly, overall survival was significantly poorer in patients with high expression of IL-22 and IL-22R than in those with low expression. These findings reveal the positive role of IL-22 and IL-22R in invasion and metastasis in human PDAC. IL-22 and IL-22R may be suitable independent prognostic markers in PDAC.

  13. Effects of BST and high energy diet on gene expression in mammary parenchyma of dairy heifers

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    Betina Joyce Lew

    2013-07-01

    Full Text Available The objective of this study was to determine the effects of dietary energy and recombinant bovine somatotropin (bST injection to identify genes that might control mammogenesis. Total RNA was extracted from the parenchymal tissue of 32 heifers randomly assigned to one of four treatments: two diets (a standard diet and a high energy, high protein diet, each with or without bST. To perform microarray experiments, RNA samples were pooled (2 animals/pool before reverse transcription and labeling with Cy3 or Cy5. A 4-node loop design was used to examine the differential gene expression among treatments using a bovine-specific cDNA microarray (National Bovine Functional Genomics Consortium Library, NBFGC containing 18,263 unique expressed sequence tags (EST. Significance levels of differential gene expression among treatments were assessed using a mixed model approach. Injection of bST altered the expression of 12 % of the genes on NBFGC slide related to tissue development, whereas 6% were altered by diet. Administration of bST increases the expression of genes positively related to cell proliferation and mammary parenchyma to a greater extent than a high energy diet.

  14. Rachiplusia nu larva as a biofactory to achieve high level expression of horseradish peroxidase.

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    Romero, Lucía Virginia; Targovnik, Alexandra Marisa; Wolman, Federico Javier; Cascone, Osvaldo; Miranda, María Victoria

    2011-05-01

    A process based on orally-infected Rachiplusia nu larvae as biological factories for expression and one-step purification of horseradish peroxidase isozyme C (HRP-C) is described. The process allows obtaining high levels of pure HRP-C by membrane chromatography purification. The introduction of the partial polyhedrin homology sequence element in the target gene increased HRP-C expression level by 2.8-fold whereas it increased 1.8-fold when the larvae were reared at 27 °C instead of at 24 °C, summing up a 4.6-fold overall increase in the expression level. Additionally, HRP-C purification by membrane chromatography at a high flow rate greatly increase D the productivity without affecting the resolution. The V(max) and K(m) values of the recombinant HRP-C were similar to those of the HRP from Armoracia rusticana roots.

  15. Screening differentially expressed genes in mouse hepatocarcinoma ascites cell line with high potential of lymphatic metastasis

    Institute of Scientific and Technical Information of China (English)

    Xiao-Nan Cui; Jian-Wu Tang; Li Hou; Bo Song; Li Li; Ji-Wei Liu

    2005-01-01

    AIM: To screen genes differentially expressed in mouse hepatocarcinoma ascites cell line with high potential of lymphatic metastasis.METHODS: A subtracted cDNA library of mouse hepatocarcinoma cell line with high potential of lymphatic metastatic Hca-F and its synogenetic cell line Hca-P with a low metastatic potential was constructed by suppression subtracted hybridization(SSH) method. The screened clones of the subtracted library were sequenced and GeneBank homology search was performed.RESULTS: Fourteen differentially expressed cDNA fragments of Hca-F were obtained with two novel genes.CONCLUSION: SSH is a useful technique to detect differentially expressioned genes and an effective method to clone novel genes.

  16. Phenomenology and Sociological Research: The Constitution of “Friendship”

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    Jochen Dreher

    2016-01-01

    Full Text Available This analysis combines phenomenological and sociological research for an investigation of the social relationship of “friendship”. From a phenomenological perspective, it focuses on the epistemological basis, on the subjective constitution of the phenomenon of “friendship”. The social construction of friendship however, is described from a sociological research perspective with reference to concrete empirical expressions of this specific social relationship. The key for the reflections on friendship is the adaptation of Edmund Husserl’s method of phenomenological reduction for the analysis of social phenomena, based on the argumentations of Alfred Schutz and Aron Gurwitsch. This leads to the assumption that friendship is symbolically constituted with reference to everyday transcendent ideas which substantially define the specific expression of this social relationship.

  17. 高温高压用耐热钢弹性蠕变损伤本构模型研究%Research on Constitutive Model of Elastic Creep Damage of Heat Resistant Steel under the condition of High temperature and High Pressure

    Institute of Scientific and Technical Information of China (English)

    马兆纬

    2001-01-01

    In the light of the fact that elastic creep damage happens to heat resistant steel used as pressure elements like steam pipeline and steam pockets in heat-engine plants at high temperature and under high pressure,this article gives a complete constitutive description of the damage,building up the theory of phenomenology about creep damage.It dwells here upon the constitutive model of tlastic creep damage of heat resistant steel,establishing a corresponding variation principle and finite element discretization form,thus introducing complete numerical variational methods.%针对火电厂中蒸汽管道与汽包类承压部件用的耐热钢,在承受高温高压时发生弹性蠕变损伤问题,给出完整的损伤本构描述,建立起唯象学蠕变损伤理论。主要提出耐热钢的弹性蠕变损伤本构模型,建立起相应的数值变分原理以及有限元离散化形式,从而形成完整的数值变分方法。

  18. High Glucose Represses hERG K+ Channel Expression through Trafficking Inhibition

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    Yuan-Qi Shi

    2015-08-01

    Full Text Available Background/Aims: Abnormal QT prolongation is the most prominent cardiac electrical disturbance in patients with diabetes mellitus (DM. It is well known that the human ether-ago-go-related gene (hERG controls the rapid delayed rectifier K+ current (IKr in cardiac cells. The expression of the hERG channel is severely down-regulated in diabetic hearts, and this down-regulation is a critical contributor to the slowing of repolarization and QT prolongation. However, the intracellular mechanisms underlying the diabetes-induced hERG deficiency remain unknown. Methods: The expression of the hERG channel was assessed via western blot analysis, and the hERG current was detected with a patch-clamp technique. Results: The results of our study revealed that the expression of the hERG protein and the hERG current were substantially decreased in high-glucose-treated hERG-HEK cells. Moreover, we demonstrated that the high-glucose-mediated damage to the hERG channel depended on the down-regulation of protein levels but not the alteration of channel kinetics. These discoveries indicated that high glucose likely disrupted hERG channel trafficking. From the western blot and immunoprecipitation analyses, we found that high glucose induced trafficking inhibition through an effect on the expression of Hsp90 and its interaction with hERG. Furthermore, the high-glucose-induced inhibition of hERG channel trafficking could activate the unfolded protein response (UPR by up-regulating the expression levels of activating transcription factor-6 (ATF-6 and the ER chaperone protein calnexin. In addition, we demonstrated that 100 nM insulin up-regulated the expression of the hERG channel and rescued the hERG channel repression caused by high glucose. Conclusion: The results of our study provide the first evidence of a high-glucose-induced hERG channel deficiency resulting from the inhibition of channel trafficking. Furthermore, insulin promotes the expression of the hERG channel

  19. Differential gene expression in patients with anal fistula reveals high levels of prolactin recepetor

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    Song Yi-Huan

    2017-01-01

    Full Text Available Background/Aim. There are limited data examining variations in the local expression of inflammatory mediators in anal fistulas where it is anticipated that an improved understanding of the inflammatory milieu might lead to the potential therapeutic option of instillation therapy in complicated cases. The aim of the present study was to examine prolactin receptors (PRLR as inflammatory markers and to correlate their expression with both the complexity of anal fistulas and the likelihood of fistula recurrence. Methods. Microarray was used to screen the differentially expressed gene profile of anal fistula using anal mucosa samples with hemorrhoids with ageand sex-matched patients as controls and then a prospective analysis of 65 patients was conducted with anal fistulas. PRLR immunohistochemistry was performed to define expression in simple, complex and recurrent anal fistula cases. The quantitative image comparison was performed combining staining intensity with cellular distribution in order to create high and low score PRLR immunohistochemical groupings. Results. A differential expression profile of 190 genes was found. PRLR expression was 2.91 times lower in anal fistula compared with control. Sixty-five patients were assessed (35 simple, 30 complex cases. Simple fistulas showed significantly higher PRLR expression than complex cases with recurrent fistulae showing overall lower PRLR expression than de novo cases (p = 0.001. These findings were reflected in measurable integrated optical density for complex and recurrent cases (complex cases, 8.31 ± 4.91 x 104 vs simple cases, 12.30 ± 6.91 x 104; p < 0.01; recurrent cases, 7.21 ± 3.51 x 104 vs primarily healing cases, 8.31 ± 4.91 x 104; p < 0.05. In univariate regression analysis, low PRLR expression correlated with fistula complexity; a significant independent effect maintained in multivariate analysis odds ratio [(OR low to high PRLR expression = 9.52; p = 0.001]. Conclusion. PRLR

  20. Propofol inhibits high glucose-induced PP2A expression in human umbilical vein endothelial cells.

    Science.gov (United States)

    Wu, Qichao; Zhao, Yanjun; Duan, Wenming; Liu, Yi; Chen, Xiangyuan; Zhu, Minmin

    2017-04-01

    Perioperative hyperglycemia is a common clinical metabolic disorder. Hyperglycemia could induce endothelial apoptosis, dysfunction and inflammation, resulting in endothelial injury. Propofol is a widely used anesthetic drug in clinical settings. Our previous studies indicated that propofol, via inhibiting high glucose-induced phosphatase A2 (PP2A) expression, attenuated high glucose-induced reactive oxygen species (ROS) accumulation, thus improving endothelial apoptosis, dysfunction and inflammation. However, the mechanisms by which propofol attenuated high glucose-induced PP2A expression is still obscure. In the present study, we examined how propofol attenuates high glucose-induced endothelial PP2A expression. Compared with 5mM glucose treatment, 15mM glucose up-regulated expression and activity of PP2A, increased cAMP response element binding protein (CREB), Ca(2+)-calmodulin dependent kinase II (CaMK II) phosphorylation and Ca(2+) accumulation. More importantly, propofol decreased PP2A expression and activity, attenuated CREB, CaMK II phosphorylation and Ca(2+) accumulation in a concentration-dependent manner. Moreover, we demonstrated that the effect of propofol was similar to that of MK801, an inhibitor of NMDA receptor. In contrast, rapastinel, an activator of NMDA receptor, antagonized the effect of propofol. Also, the effect of KN93, an inhibitor of CaMK II, was similar to that of propofol, except KN93 had no effect on 15mM glucose-mediated Ca(2+) accumulation. Our data indicated that propofol, via inhibiting NMDA receptor, attenuated 15mM glucose-induced Ca(2+) accumulation, CaMK II and CREB phosphorylation, thus inhibiting PP2A