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1

Libraries and the Internet, Part I: Some Constitutional Background.  

ERIC Educational Resources Information Center

The availability of Internet access in public libraries has raised concerns about free speech rights and library liabilities. This article, part one in a three part series, examines the constitutional background for free speech and Internet provision, defining "speech" and highlighting limitations to free speech and court analysis of speech…

Allison, Rinda Y.; Uhler, Scott F.

1997-01-01

2

Tuning constitutive recombinant gene expression in Lactobacillus plantarum.  

PubMed

Background Lactobacillus plantarum constitutes a well-recognized food-grade system for the expression of recombinant proteins in the field of industrial and medical biotechnology. For applications in vivo or in biotechnological processes, the level of expression of e.g. antigens or enzymes is often critical, as expression levels should be of a certain effectiveness, yet, without putting too much strain to the overall system. The key factors that control gene expression are promoter strength, gene copy number and translation efficiency. In order to estimate the impact of these adjusting screws in L. plantarum CD033, we have tested several constitutive promoters in combination with high and low copy number plasmid backbones and varying space between the Shine-Dalgarno sequence and the start-codon.ResultsBy combining strong promoters, such as transcription elongation factor promoters, isolated from L. plantarum CD033 and L. buchneri CD034, a synthetic promoter, originally derived from L. plantarum WCSF1 and a heterologous promoter derived from L. buchneri CD034 with a high and a low copy number origin of replication we demonstrated various expression levels of the model protein mCherry. All promoters were feasible for protein expression and in all cases, the high copy number origin of replication increased expression twofold. We found that the optimal spacer between the Shine-Dalgarno sequence and the start codon in L. plantarum consists of 8 nucleotides and elongation as well as shortening this sequence gradually down-regulates gene expression.ConclusionsWe have evaluated the effects of a set of gene regulatory tools to fine tune recombinant gene expression in L. plantarum CD033. We have thus, provided potential expression vectors useful for constitutive protein expression in lactic acid bacteria ranging from moderate to strong production levels. PMID:25410118

Tauer, Christopher; Heinl, Stefan; Egger, Esther; Heiss, Silvia; Grabherr, Reingard

2014-11-20

3

Constitutive expression of hepatic cytochrome P450 genes  

Microsoft Academic Search

One of the more challenging areas for study of the regulation of P450s is understanding the constitutive regulation of hepatic P450s. In this article, Gonzalez and Lee provide insight into how unique tissue-en- riched transcription factors regulate expression of many of the hepatic P450s that are not under direct control of the \\

FRANK J. GONZALEZ; YING-HUE LEE

1996-01-01

4

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

Microsoft Academic Search

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 lines (n=54) and a node-negative breast cancer cohort that had not received adjuvant systemic treatment (n=122). MATERIALS AND METHODS:

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

2011-01-01

5

Fish granulocytes express a constitutively active androgen receptor variant.  

PubMed

In humans, alternative splicing of androgen receptor (AR) is usually involved in some diseases. However, our knowledge about the presence of AR variants in other species and its importance for immunity is scant. Here, we report the identification of a constitutively active AR variant lacking the ligand-binding domain (LBD), AR?LBD, in the fish gilthead seabream. AR?LBD is expressed in the testis and the head-kidney (HK), and its expression varies with the reproductive stage and is correlated with plasma testosterone (T). In addition, AR?LBD is expressed in acidophilic granulocytes (AGs), which are the functional equivalent of mammalian neutrophils, but not in macrophages, and its expression is modulated by both T and immune stimuli. Notably, AR and AR?LBD were able to interact, being the activity of AR dominant at all concentrations tested of the ligand. These results reveal a new mechanism for the regulation of neutrophil biology in vertebrates and explain the conflicting results that suggest that androgens are less important than AR in human and mouse neutrophil homeostasis. PMID:24509065

Sánchez-Hernández, Miriam; Arizcun, Marta; García-Alcázar, Alicia; Sarropoulou, Elena; Mulero, Victoriano; García-Ayala, Alfonsa

2014-07-01

6

Regulated and constitutive expression of anti-inflammatory cytokines by nontransforming herpesvirus saimiri vectors.  

PubMed

Herpesviral saimiri-(HVS) mediated expression of bovine growth hormone was one of the first applications of an episomal viral vector for gene therapy. Meanwhile, the long-term persistence of HVS vectors has been confirmed in a broad spectrum of infectable target cells in vitro and in vivo. Regulated gene expression is useful for many applications of gene therapy. Therefore, we inserted the Mifepristone-antiprogestin-inducible expression system (GeneSwitchtrade mark) into HVS viral vectors to regulate the combined expression of anti-inflammatory cytokines, IL-10 and IL-1RA. Constitutive CMV-promoter/enhancer-driven and Mifepristone-inducible cytokine expression was compared in the viral context in transduced primary human fibroblasts and rheumatoid arthritis (RA) fibroblast-like cells (RASF). Long-term persistence of vector genomes was shown for both construct types. Constitutive expression was efficient and more rapid in onset than in the inducible system, in which the selective induction of interleukin expression along with low background levels was obtained by Mifepristone concentrations that were more than 1000-fold below those required for endogenous Progesterone antagonism. Furthermore, transgene expression corresponded to vector doses. Global patterns of cytokine secretion were not significantly changed due to viral transduction, indicating a rather inert behavior of the viral vector itself. In an attempt to emulate the inflammatory cytokine-enriched environment in rheumatoid arthritic joints, the function of the vectors could be demonstrated in vitro by the successful blockade of IL-1beta-stimulated matrix-metalloproteinase (MMP)-3 expression from RASF cells. Evaluation of this system in future studies, in suitable long-term SCID models of RA or in non-human primate models, will exploit the possible in vivo benefits of nontransforming HVS vectors in gene therapy. PMID:15647772

Wieser, C; Stumpf, D; Grillhösl, C; Lengenfelder, D; Gay, S; Fleckenstein, B; Ensser, A

2005-03-01

7

Constitutive heterochromatin: a surprising variety of expressed sequences.  

PubMed

The organization of chromosomes into euchromatin and heterochromatin is amongst the most important and enigmatic aspects of genome evolution. Constitutive heterochromatin is a basic yet still poorly understood component of eukaryotic chromosomes, and its molecular characterization by means of standard genomic approaches is intrinsically difficult. Although recent evidence indicates that the presence of transcribed genes in constitutive heterochromatin is a conserved trait that accompanies the evolution of eukaryotic genomes, the term heterochromatin is still considered by many as synonymous of gene silencing. In this paper, we comprehensively review data that provide a clearer picture of transcribed sequences within constitutive heterochromatin, with a special emphasis on Drosophila and humans. PMID:19412619

Dimitri, Patrizio; Caizzi, Ruggiero; Giordano, Ennio; Carmela Accardo, Maria; Lattanzi, Giovanna; Biamonti, Giuseppe

2009-08-01

8

Downregulation of Constitutive and Cytokine-Induced Complement 3 Expression by Morphine in Rat Astrocytes  

PubMed Central

Background The effect of opioids on inflammation and immune responses is an important subject of investigation because immunoregulatory cytokines are produced in the central nervous system and opioid receptors are widespread in these cells. Objectives The aim of this study was to evaluate the immunomodulatory effect of morphine on the C3 expression (both constitutive and proinflammatory cytokine-induced C3 expression) in primary rat astrocytes. Methods Primary rat astrocytes were untreated or treated with morphine in different concentrations (10–6 to 10–2 M) before incubation without or with 5 U/mL tumor necrosis factor-? (TNF-?), and C3 protein and mRNA expressions were measured. Similarly, astrocytes were treated with 10–3 M morphine and stimulated with other proinflammatory cytokines, including 10 ng/mL interleukin-8 (IL-8) and 5 U/mL IL-1?. Astrocytes were exposed to 10–5 M naloxone for 2 hours before adding morphine, and TNF-? and C3 protein was measured. Tumor growth factor-? (TGF-?) was measured from the supernatants of each proinflammatory cytokine. Results All results are expressed as mean percentages of C3 production by normalizing C3 without morphine or any cytokine treatment as 100%. Constitutive C3 protein production was decreased at morphine 10–3 M (57.2%) and 10–2 M (30.1%). Pretreatment with morphine suppressed induction of C3 expression at both the protein and mRNA levels in astrocytes stimulated with TNF-?, IL-8, and IL-1? (P < 0.05) in a dose-dependent manner. The inhibition of C3 protein production by morphine (10–3 M; 33%) was partially attenuated by naloxone (52.0%) (P < 0.05). The pretreatment of astrocytes with morphine (10–3 M) before stimulation with TNF-?, IL-8, and IL-1? increased by 33% (P < 0.05), decreased by 15.2% (P < 0.05), and did not change the production of TGF-? protein, respectively. Conclusions Morphine downregulated both constitutive and proinflammatory cytokine-induced C3 expression of astrocytes at the transcriptional level, but not in a cytokine-specific manner. PMID:24648573

Kim, Chung Su; Ko, Justin Sangwook; Lee, Ae Ryoung; Shin, Byung Seop; Choi, Soo Joo; Lee, Jeong Jin; Kim, Hyun Soo; Lee, Sangmin Maria

2011-01-01

9

Constitutive versus Responsive Gene Expression Strategies for Growth in Changing Environments  

PubMed Central

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 to verify them. PMID:22140435

Geisel, Nico

2011-01-01

10

Constitutive expression of ciliary neurotrophic factor in mouse hypothalamus  

PubMed Central

Ciliary neurotrophic factor (CNTF) is a potent survival molecule for a large number of neuronal and glial cells in culture; its expression in glial cells is strongly upregulated after a variety of nerve tissue injuries. Exogenously administered CNTF produces an anorectic effect via activation of hypothalamic neurons and stimulates neurogenesis in mouse hypothalamus. To determine whether CNTF is produced endogenously in the hypothalamus, we sought cellular sources and examined their distribution in adult mouse hypothalamus by immunohistochemistry. CNTF immunoreactivity (IR) was predominantly detected in the ependymal layer throughout the rostrocaudal extension of the third ventricle, where numerous ependymocytes and tanycytes exhibited specific staining. Some astrocytes in the grey matter of the anterior hypothalamus and in the median eminence of the hypothalamic tuberal region were also positive. Stimulation of cells bearing CNTF receptor ? (CNTFR?) induces specific activation of the signal transducer and activator of transcription 3 (STAT3) signalling system. Treatment with recombinant CNTF and detection of the nuclear expression of phospho-STAT3 (P-STAT3) showed that CNTF-producing ependymal cells and tanycytes were intermingled with, or very close to, P-STAT3-positive, CNTFR?-bearing cells. A fraction of CNTF-producing ependymal cells and tanycytes and some median eminence astrocytes also exhibited P-STAT3 IR. Thus, in normal adult mice the ependyma of the third ventricle is both a source of and a target for CNTF, which may play hitherto unknown roles in hypothalamic function in physiological conditions. PMID:22458546

Severi, Ilenia; Carradori, Maria Rita; Lorenzi, Teresa; Amici, Adolfo; Cinti, Saverio; Giordano, Antonio

2012-01-01

11

Construction of a plasmid for heterologous protein expression with a constitutive promoter in Trichoderma reesei.  

PubMed

Trichoderma reesei is widely used as a host for homologus and heterologus protein expression because of its well-known capability of protein secretion, especially cellulases secretion. In this study, a binary vector which could be used for protein expression was constructed with a constitutive promoter of rp2 gene. This vector contained the expression cassette Prp2-target gene-Trp2 and hygromycin B selection marker based on pCAMBIA1300 for T-DNA random insertion. The feasibility of the promoter was determined by eGFP (enhanced green fluorescence protein) expression in T. reesei. For heterologus protein expression, the expression of bgla from Aspergillus niger in the transformant was 3 folds higher than that of the wild type strain. The results demonstrate that this constitutive promoter could be applied for protein expression and thus this protein express system may contribute to the industrial protein production. PMID:24120481

He, Ronglin; Zhang, Can; Guo, Wei; Wang, Lixian; Zhang, Dongyuan; Chen, Shunlin

2013-11-01

12

Defective neuromuscular synaptogenesis in mice expressing constitutively active ErbB2 in skeletal muscle fibers  

Microsoft Academic Search

We overexpressed a constitutively active form of the neuregulin receptor ErbB2 (CAErbB2) in skeletal muscle fibers in vivo and in vitro by tetracycline-inducible expression. Surprisingly, CAErbB2 expression during embryonic development was lethal and impaired synaptogenesis yielding a phenotype with loss of synaptic contacts, extensive axonal sprouting, and diffuse distribution of acetylcholine receptor (AChR) transcripts, reminiscent of agrin-deficient mice. CAErbB2 expression

Olga N. Ponomareva; Hualong Ma; Vita M. Vock; Elaine L. Ellerton; Susan E. Moody; Ramzi Dakour; Lewis A. Chodosh; Mendell Rimer

2006-01-01

13

Stable expression of constitutively-activated STAT3 in benign prostatic epithelial cells changes their phenotype to that resembling malignant cells  

Microsoft Academic Search

BACKGROUND: Signal transducers and activators of transcription (STATs) are involved in growth regulation of cells. They are usually activated by phosphorylation at specific tyrosine residues. In neoplastic cells, constitutive activation of STATs accompanies growth dysregulation and resistance to apoptosis through changes in gene expression, such as enhanced anti-apoptotic gene expression or reduced pro-apoptotic gene expression. Activated STAT3 is thought to

Hosea F Huang; Thomas F Murphy; Ping Shu; Arnold B Barton; Beverly E Barton

2005-01-01

14

Constitutive homologous expression of phosphoglucomutase and transaldolase increases the metabolic flux of Fusarium oxysporum  

PubMed Central

Background Fusarium oxysporum is among the few filamentous fungi that have been reported of being able to directly ferment biomass to ethanol in a consolidated bioprocess. Understanding its metabolic pathways and their limitations can provide some insights on the genetic modifications required to enhance its growth and subsequent fermentation capability. In this study, we investigated the hypothesis reported previously that phosphoglucomutase and transaldolase are metabolic bottlenecks in the glycolysis and pentose phosphate pathway of the F. oxysporum metabolism. Results Both enzymes were homologously overexpressed in F. oxysporum F3 using the gpdA promoter of Aspergillus nidulans for constitutive expression. Transformants were screened for their phosphoglucomutase and transaldolase genes expression levels with northern blot. The selected transformant exhibited high mRNA levels for both genes, as well as higher specific activities of the corresponding enzymes, compared to the wild type. It also displayed more than 20 and 15% higher specific growth rate upon aerobic growth on glucose and xylose, respectively, as carbon sources and 30% higher biomass to xylose yield. The determination of the relative intracellular amino and non-amino organic acid concentrations at the end of growth on glucose revealed higher abundance of most determined metabolites between 1.5- and 3-times in the recombinant strain compared to the wild type. Lower abundance of the determined metabolites of the Krebs cycle and an 68-fold more glutamate were observed at the end of the cultivation, when xylose was used as carbon source. Conclusions Homologous overexpression of phosphoglucomutase and transaldolase in F. oxysporum was shown to enhance the growth characteristics of the strain in both xylose and glucose in aerobic conditions. The intracellular metabolites profile indicated how the changes in the metabolome could have resulted in the observed growth characteristics. PMID:24649884

2014-01-01

15

[Inducible and constitutive expression of fip-fve from Flammulina velutipes in Pichia pastoris GS115].  

PubMed

We transformed the fip-fve gene into Pichia pastoris GS115 for inducible and constitutive expression to obtain feasible bioactvie recombinant Fip-fve. The fip-fve gene was cloned from Flammulina velutipes fruting body by PCR and ligated to pPIC9 to construct inducible expression vector pPIC9-FIP-fve, and promotor pgap was used to replace the paox1 to construct constitutive expression vector pPIC9-PGAP-FIP-fve. These two vectors were used to transform P. pastoris by PEG method. The fip-fve was expressed after histamine-absence screening and yeast colony PCR. The inducible expression level reached 158.2 mg/L at the fourth day and the constitutive expression level was 46.3 mg/L and 29.5 mg/L using glucose and glycerol, respectively. The SDS-PAGE and Western blotting both proved the correctness of rFip-fve, and the hemagglutination test indicats the rFip-fve's bioactivity. PMID:25007582

Lin, Jingwei; Jia, Jia; Zhong, Ming; Chen, Lijing; Li, Haoge; Guo, Zhifu; Qi, Mingfang; Liu, Lixia; Li, Tianlai

2014-03-01

16

A soybean (Glycine max) polyubiquitin promoter gives strong constitutive expression in transgenic soybean.  

PubMed

The success of plant genetic transformation relies greatly on the strength and specificity of the promoters used to drive genes of interest. In this study, we analyzed gfp gene expression mediated by a polyubiquitin promoter (Gmubi) from soybean (Glycine max) in stably transformed soybean tissues. Strong GFP expression was observed in stably transformed proliferative embryogenic tissues. In whole transgenic plants, GFP expression was observed in root tips, main and lateral roots, cotyledons and plumules in young plants as well as in leaf veins, petioles, flower petals, pollen, pods and developing seeds in mature plants. GFP expression was localized mainly in epidermal cells, leaf mesophyll, procambium and vascular tissues. Introduction of an intron-less version of the Gmubi promoter (Gmupri) displayed almost the same GFP expression pattern albeit at lower intensities. The Gmubi promoter showed high levels of constitutive expression and represents an alternative to viral promoters for driving gene expression in soybean. PMID:19229538

Hernandez-Garcia, Carlos M; Martinelli, Adriana P; Bouchard, Robert A; Finer, John J

2009-05-01

17

High Level Constitutive Expression of Luciferase Reporter by lsd90 Promoter in Fission Yeast  

PubMed Central

Because of a large number of molecular similarities with higher eukaryotes, the fission yeast Schizosaccharomyces pombe has been considered a potentially ideal host for expressing human proteins having therapeutic and pharmaceutical applications. However, efforts in this direction are hampered by lack of a strong promoter. Here, we report the isolation and characterization of a strong, constitutive promoter from S. pombe. A new expression vector was constructed by cloning the putative promoter region of the lsd90 gene (earlier reported to be strongly induced by heat stress) into a previously reported high copy number vector pJH5, which contained an ARS element corresponding to the mat2P flanking region and a truncated URA3m selectable marker. The resulting vector was used to study and compare the level of expression of the luciferase reporter with that achieved with the known vectors containing regulatable promoter nmt1 and the strong constitutive promoter adh1 in S. pombe and the methanol-inducible AOX1 promoter in Pichia pastoris. Following growth in standard media the new vector containing the putative lsd90 promoter provided constitutive expression of luciferase, at a level, which was 19-, 39- and 10-fold higher than that achieved with nmt1, adh1 and AOX1 promoters, respectively. These results indicate a great potential of the new lsd90 promoter-based vector for commercial scale expression of therapeutic proteins in S. pombe. PMID:24999979

Verma, Hemant Kumar; Shukla, Poonam; Alfatah, Md.; Khare, Asheesh Kumar; Upadhyay, Udita; Ganesan, Kaliannan; Singh, Jagmohan

2014-01-01

18

Transgenic Expression of Constitutively Active RAC1 Disrupts Mouse Rod Morphogenesis  

PubMed Central

Purpose. Dominant-active RAC1 rescues photoreceptor structure in Drosophila rhodopsin-null mutants, indicating an important role in morphogenesis. This report assesses the morphogenetic effect of activated RAC1 during mammalian rod photoreceptor development using transgenic mice that express constitutively active (CA) RAC1. Methods. Transgenic mice were generated by expressing CA RAC1 under control of the Rhodopsin promoter, and morphological features of the photoreceptors were evaluated by histology, immunohistochemistry, and transmission electron microscopy. Function was evaluated by electroretinography. Potential protein partners of CA RAC1 were identified by co-immunoprecipitation of retinal extracts. Results. Constitutively active RAC1 expression in differentiating rods disrupted outer retinal lamination as early as postnatal day (P)6, and many photoreceptor cell nuclei were displaced apically into the presumptive subretinal space. These photoreceptors did not develop normal inner and outer segments and had abnormal placement of synaptic elements. Some photoreceptor nuclei were also mislocalized into the inner nuclear layer. Extensive photoreceptor degeneration was subsequently observed in the adult animal. Constitutively active RAC1 formed a complex with the polarity protein PAR6 and with microtubule motor dynein in mouse retina. The normal localization of the PAR6 complex was disrupted in CA RAC1-expressing rod photoreceptors. Conclusions. Constitutively active RAC1 had a profound negative effect on mouse rod cell viability and development. Rod photoreceptors in the CA RAC1 retina exhibited a defect in polarity and migration. Constitutively active RAC1 disrupted rod morphogenesis and gave a phenotype resembling that found in the Crumbs mutant. PAR6 and dynein are two potential downstream effectors that may be involved in CA RAC1-mediated defective mouse photoreceptor morphogenesis. PMID:24651551

Song, Hongman; Bush, Ronald A.; Vijayasarathy, Camasamudram; Fariss, Robert N.; Kjellstrom, Sten; Sieving, Paul A.

2014-01-01

19

Constitutive Expression of Human Bcl2 Modulates Nitrogen Mustard and Camptothecin Induced Apoptosis  

Microsoft Academic Search

Bcl-2 is a novel protooncogene which prolongs cell survival and sup presses apoptosis. We examined whether constitutive expression of trans- fected human bcl-2 conferred resistance to two different DNA damaging drugs, nitrogen mustard (HN2) and camptothecin (CPT) in a murine, IL-3 dependent cell line (FL5.12). HN2 treatment produced 2-fold less cell death and DNA degradation in cells overexpressing bcl-2 relative

M. I. Walton; D. Whysong; P. M. O'Connor; D. Hockenbery; S. J. Korsmeyer; K. W. Kohn

1993-01-01

20

Malignant transformation of human cells by constitutive expression of platelet-derived growth factor-BB.  

PubMed

Platelet-derived growth factors (PDGFs) comprise a family of growth factors strongly implicated in human oncogenesis. A number of human tumors overexpress PDGF family members or have translocations activating PDGF receptors. Whereas the epidemiologic evidence implicating PDGF in human tumors is strong, malignant transformation of human cells by overexpression of PDGF has not been demonstrated. We have previously developed a human cell line by the sequential introduction of large T cells and telomerase, and we have demonstrated that these cells express functionally active PDGF receptor (PDGFR) beta. In order to determine whether growth factor-mediated transformation of human cells could occur, these cells were transduced with a retrovirus encoding PDGF-BB. Constitutive expression of PDGF-BB led to malignant transformation in nude mice. This is the first demonstration of constitutive signaling causing malignant transformation of human cells. Some of the changes that occur because of constitutive growth factor expression can be reversed by the clinically approved tyrosine kinase inhibitor Glivec, whereas other changes are not reversible by tyrosine kinase inhibitors. Our model allows the assessment of epigenetic changes that occur during human carcinogenesis. In addition, these studies provide insight into the clinical failure of tyrosine kinase inhibitors as monotherapy for advanced malignancy. PMID:15695519

Govindarajan, Baskaran; Shah, Asha; Cohen, Cynthia; Arnold, Rebecca S; Schechner, Jeffrey; Chung, Jun; Mercurio, Arthur M; Alani, Rhoda; Ryu, Byungwoo; Fan, Chun-Yang; Cuezva, Jose M; Martinez, Marta; Arbiser, Jack L

2005-04-01

21

Constitutive expression of short hairpin RNA in vivo triggers buildup of mature hairpin molecules.  

PubMed

RNA interference (RNAi) has become the cornerstone technology for studying gene function in mammalian cells. In addition, it is a promising therapeutic treatment for multiple human diseases. Virus-mediated constitutive expression of short hairpin RNA (shRNA) has the potential to provide a permanent source of silencing molecules to tissues, and it is being devised as a strategy for the treatment of liver conditions such as hepatitis B and hepatitis C virus infection. Unintended interaction between silencing molecules and cellular components, leading to toxic effects, has been described in vitro. Despite the enormous interest in using the RNAi technology for in vivo applications, little is known about the safety of constitutively expressing shRNA for multiple weeks. Here we report the effects of in vivo shRNA expression, using helper-dependent adenoviral vectors. We show that gene-specific knockdown is maintained for at least 6 weeks after injection of 1?×?10(11) viral particles. Nonetheless, accumulation of mature shRNA molecules was observed up to weeks 3 and 4, and then declined gradually, suggesting the buildup of mature shRNA molecules induced cell death with concomitant loss of viral DNA and shRNA expression. No evidence of well-characterized innate immunity activation (such as interferon production) or saturation of the exportin-5 pathway was observed. Overall, our data suggest constitutive expression of shRNA results in accumulation of mature shRNA molecules, inducing cellular toxicity at late time points, despite the presence of gene silencing. PMID:21780944

Ahn, M; Witting, S R; Ruiz, R; Saxena, R; Morral, Núria

2011-12-01

22

CT-2576, an inhibitor of phospholipid signaling, suppresses constitutive and induced expression of human immunodeficiency virus.  

PubMed Central

Viruses such as human immunodeficiency virus (HIV) require cellular activation for expression. Cellular activation in lymphoid cells is associated with augmented accumulation of certain phosphatidic acid (PA) species derived from the hydrolysis of glycan phosphatidylinositol (GPI). This suggests that activation of a phospholipid pathway may play a role in initiation of viral replication. To test this hypothesis, we examined the effect of tat gene expression on the production of cellular PA species, as the Tat protein is essential for HIV expression and has been implicated in activating the expression of multiple host cellular genes. Expression of tat increased the expression of PA. We then tested whether synthetic inhibitors of PA metabolism would inhibit activation of the HIV long terminal repeat by Tat and tumor necrosis factor alpha (TNF-alpha). CT-2576 suppressed both PA generation induced by Tat and HIV long terminal repeat-directed gene expression in response to Tat or TNF-alpha at a posttranscriptional step. CT-2576 also inhibited constitutive as well as TNF-alpha- and interleukin 6-induced expression of HIV p24 antigen in chronically infected U1 cells and in peripheral blood lymphocytes acutely infected with a clinical isolate of HIV. Pharmacological inhibition of synthesis of selected PA species may therefore provide a therapeutic approach to suppression of HIV replication. Images Fig. 1 Fig. 3 Fig. 5 PMID:7761405

Leung, D W; Peterson, P K; Weeks, R; Gekker, G; Chao, C C; Kaplan, A H; Balantac, N; Tompkins, C; Underiner, G E; Bursten, S

1995-01-01

23

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

PubMed Central

Background The Biopeptide BP100 is a synthetic and strongly cationic ?-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 ?-helical synthetic peptides can have a strong negative impact on rice fitness. However, GM plants expressing, for example, BP100 based on inverted repeats, have adequate agronomic performance and resistant phenotypes as a result of a complex equilibrium between bp100der toxicity to plant cells, antimicrobial activity and transgene-derived plant stress response. It is likely that these results can be extended to other peptides with similar characteristics. PMID:22947243

2012-01-01

24

Enhancement of cellulose pellicle production by constitutively expressing vitreoscilla hemoglobin in Acetobacter xylinum.  

PubMed

Vitreoscilla hemoglobin (VHb) gene driven by the constitutive bla promoter was expressed in the cellulose-producing Acetobacter xylinum. The expressed VHb was biochemically active and could enhance cell growth in a shaken culture containing cellulase. VHb-expressing A. xylinum (VHb+) exhibited a specific growth rate 50% higher than that of the host strain (VHb-). Probably because of its faster growth rate, the size of tentacled cellulose beads produced by VHb+ was about 20% of that produced by VHb- after 2 days cultivation in a shake-flask. When cultured statically, the amount of cellulose pellicle produced by VHb+ could be 2-fold that produced by VHb-. Cellulose pellicle concentration of 11 g/L was obtained for VHb+, whereas 6 g/L was obtained for VHb- after 6 days of microaerobic incubation. PMID:17137307

Chien, Liang-Jung; Chen, Hsiao-Ting; Yang, Pei-Fen; Lee, Cheng-Kang

2006-01-01

25

Constitutive and inducible expression of SKALP/elafin provides anti-elastase defense in human epithelia.  

PubMed Central

Skin-derived antileukoproteinase (SKALP), also known as elafin, is a serine proteinase inhibitor first discovered in keratinocytes from hyperproliferative human epidermis. In addition to the proteinase inhibiting domain which is directed against polymorphonuclear leukocyte (PMN) derived enzymes such as elastase and proteinase 3, SKALP contains multiple transglutaminase (TGase) substrate domains which enable crosslinking to extracellular and cell envelope proteins. Here we show that SKALP is constitutively expressed in several epithelia that are continuously subjected to inflammatory stimuli, such as the oral cavity and the vagina where it co-localizes with type 1 TGase. All epithelia from sterile body cavities are negative for SKALP. In general, stratified squamous epithelia are positive, whereas pseudostratified epithelia, simple/glandular epithelia and normal epidermis are negative. SKALP was found in fetal tissues of the oral cavity from 17 wk gestation onwards where it continued to be expressed up to adult life. Remarkably, in fetal epidermis SKALP was found from week 28 onwards, but was downregulated to undetectable levels in neonatal skin within three months, suggesting a role during pregnancy in feto-maternal interactions or in the early maturation phase of the epidermis. Immunoelectron microscopy revealed the presence of SKALP in secretory vesicles including the lamellar granules. In culture models for epidermal keratinocytes we found that expression of the endogenous SKALP gene provided protection against cell detachment caused by purified elastase or activated PMNs. Addition of exogenous recombinant SKALP fully protected the keratinocytes against PMN-dependent detachment whereas superoxide dismutase and catalase were only marginally effective. These findings strongly suggest that the constitutive expression of SKALP in squamous epithelia, and the inducible expression in epidermis participate in the control of epithelial integrity, by inhibiting PMN derived proteinases. PMID:8823304

Pfundt, R; van Ruissen, F; van Vlijmen-Willems, I M; Alkemade, H A; Zeeuwen, P L; Jap, P H; Dijkman, H; Fransen, J; Croes, H; van Erp, P E; Schalkwijk, J

1996-01-01

26

Resistance to simian immunodeficiency virus low dose rectal challenge is associated with higher constitutive TRIM5? expression in PBMC  

PubMed Central

Background At least six host-encoded restriction factors (RFs), APOBEC3G, TRIM5?, tetherin, SAMHD1, schlafen 11, and Mx2 have now been shown to inhibit HIV and/or SIV replication in vitro. To determine their role in vivo in the resistance of macaques to mucosally-acquired SIV, we quantified both pre-exposure (basal) and post-exposure mRNA levels of these RFs, Mx1, and IFN? in PBMC, lymph nodes, and duodenum of rhesus macaques undergoing weekly low dose rectal exposures to the primary isolate, SIV/DeltaB670. Results Repetitive challenge divided the monkeys into two groups with respect to their susceptibility to infection: highly susceptible (2–3 challenges, 5 monkeys) and poorly susceptible (?6 challenges, 3 monkeys). Basal RF and Mx1 expression varied among the three tissues examined, with the lowest expression generally detected in duodenal tissues, and the highest observed in PBMC. The one exception was A3G whose basal expression was greatest in lymph nodes. Importantly, significantly higher basal expression of TRIM5? and Mx1 was observed in PBMC of animals more resistant to mucosal infection. Moreover, individual TRIM5? levels were stable throughout a year prior to infection. Post-exposure induction of these genes was also observed after virus appearance in plasma, with elevated levels in PBMC and duodenum transiently occurring 7–10 days post infection. They did not appear to have an effect on control of viremia. Interestingly, minimal to no induction was observed in the resistant animal that became an elite controller. Conclusions These results suggest that constitutively expressed TRIM5? appears to play a greater role in restricting mucosal transmission of SIV than that associated with type I interferon induction following virus entry. Surprisingly, this association was not observed with the other RFs. The higher basal expression of TRIM5? observed in PBMC than in duodenal tissues emphasizes the understated role of the second barrier to systemic infection involving the transport of virus from the mucosal compartment to the blood. Together, these observations provide a strong incentive for a more comprehensive examination of the intrinsic, variable control of constitutive expression of these genes in the sexual transmission of HIV. PMID:24884551

2014-01-01

27

Effect of chromosome constitution variations on the expression of Turner phenotype.  

PubMed

Turner syndrome (TS) is a chronic disease related to haploinsufficiency of genes that are normally expressed in both X chromosomes in patients with female phenotype that is associated with a wide range of somatic malformations. We made detailed cytogenetic and clinical analysis of 65 patients with TS from the region of Recife, Brazil, to determine the effects of different chromosome constitutions on expression of the TS phenotype. Overall, patients with X-monosomy exhibited a tendency to have more severe phenotypes with higher morbidity, showing its importance in TS prognosis. Additionally, we found rare genetic and phenotypic abnormalities associated with this syndrome. To the best of our knowledge, this is the first case of 45,X,t(11;12)(q22;q22) described as a TS karyotype. Turner patients usually have normal intelligence; however, moderate to severe levels of mental retardation were found in 5 TS cases, which is considerate a very uncommon feature in this syndrome. PMID:23546984

Bispo, A V S; Dos Santos, L O; Burégio-Frota, P; Galdino, M B; Duarte, A R; Leal, G F; Araújo, J; Gomes, B; Soares-Ventura, E M; Muniz, M T C; Santos, N

2013-01-01

28

A Constitutive Unregulated Expression of ?-Galactosidase in Lactobacillus fermentum M1.  

PubMed

A constitutively ?-galactosidase (LacL)-producing Lactobacillus fermentum M1 isolated from fermented milk was found to produce ?-galactosidase in the presence of glucose. ?-galactosidase activity produced in glucose (30 mM) medium was 2.17 U/mL as compared to 2.27 and 2.19 U/mL with galactose and lactose, respectively. When a combination of glucose (30 or 60 mM) with galactose (30 mM) was used as carbon source, ?-galactosidase activity was not repressed rather was found increased when compared to carbon sources used individually. In real-time PCR analysis of mRNA synthesized on individual and combined carbon sources, repression of the lacL gene expression was not observed. This observation suggests that the strain M1 lacked normal carbon catabolite repression. Examination of nucleotide sequence of lacL identified two catabolite responsive elements (cre): cre1 located downstream near the promoter region and cre2 within the coding sequence. Each of which differed from the 14-bp consensus by a single nucleotide. In cre1, it is C in place of highly conserved T at position 1 in the consensus. In cre 2, it is G in place of C, a residue completely conserved at position 13. Since catabolite genes in Gram-positive bacteria are regulated by carbon catabolite protein A (CcpA) through interaction with DNA at a specific cis-acting cre, it is assumed that base changes at conserved position in the cre elements disrupt CcpA binding and thereby leading to constitutive expression of lacL gene. The study noted to be the first report about the constitutive production of ?-galactosidase in L. fermentum. PMID:25319027

Arukha, Ananta Prasad; Mukhopadhyay, Bidhan Chandra; Mitra, Suranjita; Biswas, Swadesh Ranjan

2015-02-01

29

Constitutive expression of cell wall invertase genes increases grain yield and starch content in maize.  

PubMed

Grain size, number and starch content are important determinants of grain yield and quality. One of the most important biological processes that determine these components is the carbon partitioning during the early grain filling, which requires the function of cell wall invertase. Here, we showed the constitutive expression of cell wall invertase-encoding gene from Arabidopsis, rice (Oryza sativa) or maize (Zea mays), driven by the cauliflower mosaic virus (CaMV) 35S promoter, all increased cell wall invertase activities in different tissues and organs, including leaves and developing seeds, and substantially improved grain yield up to 145.3% in transgenic maize plants as compared to the wild-type plants, an effect that was reproduced in our 2-year field trials at different locations. The dramatically increased grain yield is due to the enlarged ears with both enhanced grain size and grain number. Constitutive expression of the invertase-encoding gene also increased total starch content up to 20% in the transgenic kernels. Our results suggest that cell wall invertase gene can be genetically engineered to improve both grain yield and grain quality in crop plants. PMID:23926950

Li, Bei; Liu, Hua; Zhang, Yue; Kang, Tao; Zhang, Li; Tong, Jianhua; Xiao, Langtao; Zhang, Hongxia

2013-12-01

30

Constitutive TL1A (TNFSF15) Expression on Lymphoid or Myeloid Cells Leads to Mild Intestinal Inflammation and Fibrosis  

PubMed Central

TL1A is a member of the TNF superfamily and its expression is increased in the mucosa of inflammatory bowel disease patients. Moreover, a subset of Crohn's disease (CD) patients with the risk TL1A haplotype is associated with elevated TL1A expression and a more severe disease course. To investigate the in vivo role of elevated TL1A expression, we generated two transgenic (Tg) murine models with constitutive Tl1a expression in either lymphoid or myeloid cells. Compared to wildtype (WT) mice, constitutive expression of Tl1a in either lymphoid or myeloid cells showed mild patchy inflammation in the small intestine, which was more prominent in the ileum. In addition, mice with constitutive Tl1a expression exhibited enhanced intestinal and colonic fibrosis compared to WT littermates. The percentage of T cells expressing the gut homing chemokine receptors CCR9 and CCR10 was higher in the Tl1a Tg mice compared to WT littermates. Sustained expression of Tl1A in T cells also lead to increased Foxp3+ Treg cells. T cells or antigen presenting cells (APC) with constitutive expression of Tl1a were found to have a more activated phenotype and mucosal mononuclear cells exhibit enhanced Th1 cytokine activity. These results indicated an important role of TL1A in mucosal T cells and APC function and showed that up-regulation of TL1A expression can promote mucosal inflammation and gut fibrosis. PMID:21264313

Shih, David Q.; Barrett, Robert; Zhang, Xiaolan; Yeager, Nicole; Koon, Hon Wai; Phaosawasdi, Piangwarin; Song, Yahui; Ko, Brian; Wong, Michelle H.; Michelsen, Kathrin S.; Martins, Gislaine; Pothoulakis, Charalabos; Targan, Stephan R.

2011-01-01

31

Constitutive and inducible expression of megakaryocyte-specific genes in Friend erythroleukaemia cells.  

PubMed

Friend murine erythroleukaemia cells (MELC) were analysed by semiquantitative RT-PCR for the constitutive and inducible expression of megakaryocyte-specific genes. Uninduced MELC expressed detectable levels of mRNAs for acethylcholinesterase (AChE), platelet factor-4 (PF4), glycoprotein IIb (GPIIb) and von Willebrand factor (VWF), whereas the erythroid alpha- and beta-globin genes were not transcribed appreciably. However, MELC exposed to 5 mM hexamethylene bisacetamide (HMBA) or 1.5% dimethyl sulphoxide (DMSO) seemed to be channelled towards a mixed erythroid/megakaryocytic phenotype characterized by unaltered levels of VWF mRNA, increased levels of AChE, GPIIb and PF4 mRNA. and simultaneous induction of the globin genes. Megakaryocyte-related genes were expressed. in the absence of globin gene transcription, by MELC treated with either phorbol-12-myristate acetate (PMA; 100 ng/ml) or colcemid (40 nM), an antimicrotubule agent capable of promoting polyploidization in this model. Moreover, PMA and colcemid induced also de novo expression of the thrombopoietin receptor c-mpl. PMA and colcemid did not affect high basal c-myb mRNA levels which, in turn, were down-regulated upon HMBA or DMSO induction. Additionally, both uninduced and induced MELC exhibited significant levels of Epo-R and IL-3R mRNAs, whereas no expression of granulocyte/macrophage-related genes was detected. Megakaryocyte gene expression of MELC was also compared to that of other haemopoietic cell populations from normal mice and mice infected with the anaemic strain of the Friend virus. According to our results, MELC should be seen as an unique erythro-megakaryocytic model of differentiation, potentially useful for studying molecular events governing lineage commitment as well as some steps of megakaryocytopoiesis. PMID:9401056

Vannucchi, A M; Linari, S; Cellai, C; Koury, M J; Paoletti, F

1997-12-01

32

Constitutive expression of three novel catfish CXC chemokines: homeostatic chemokines in teleost fish.  

PubMed

Chemokines are best known for their vital role in leukocyte chemotaxis, as part of the larger inflammatory response. Expression analysis and functional characterization of chemokines in mammalian species have often overlooked the role of these proteins under homeostatic conditions. Recent investigations of chemokine diversity in teleost fish have also centered on the immune-related functions of chemotactic cytokines, such as CXCL8 and CXCL10. While a disease-based approach to chemokines is essential to the development of remediative therapies for both human and animal infections, it may be a poor measure of the overall complexity of chemokine functions. As part of a larger effort to assess the conservation of chemokine diversity in teleost fish, we report here the identification of three novel, constitutively expressed CXC chemokines from channel catfish (Ictalurus punctatus). Phylogenetic analyses indicated that two of the three CXC chemokines were orthologues for mammalian CXCL12 and CXCL14, respectively. Whereas a clear orthology could not yet be established for the third CXC chemokine, it shared highest amino acid identity with mammalian CXCL2. All three CXC chemokines show expression in a wide range of tissues, and early expression during development was observed for CXCL12. The expression of this new set of catfish CXC chemokines was not induced during challenge by infection of Edwardsiella ictaluri, the causative agent of the fish pathogen enteric septicemia of catfish. In contrast to the gene duplication of CXCL12 in carp and zebrafish, Southern blot analysis indicated that all three catfish CXC chemokines exist as single copy genes in the catfish genome suggesting that gene duplication of CXC chemokines in specific teleost fish was a recent evolutionary event. PMID:15950731

Baoprasertkul, Puttharat; He, Chongbo; Peatman, Eric; Zhang, Shanzhong; Li, Ping; Liu, Zhanjiang

2005-07-01

33

The constitutive expression of a two transgene construct enhances the abiotic stress tolerance of chrysanthemum.  

PubMed

Various abiotic stresses downgrade the quality and productivity of chrysanthemum. A construct carrying both CcSOS1 (from Chrysanthemum crassum) and CdICE1 (from Chrysanthemum dichrum) was constitutively expressed in the chrysanthemum variety 'Jinba'. The transgenic plants were superior to the wild type (WT) ones with respect to their sensitivity to low temperature, drought and salinity, as measured by visible damage and plant survival. Salinity stressed transgenic plants accumulated more proline, and their level of superoxide dismutase and peroxidase activity was higher than in WT plants. At the physiological level, they suffered less loss of viable leaf area, maintained a lower leaf electrolyte conductivity and retained more chlorophyll (a+b). The ratio between the K(+) and Na(+) content was higher in the root, stem and median leaves of salinity stressed transgenic plants than in those of WT plants. PMID:24751398

Song, Aiping; An, Juan; Guan, Zhiyong; Jiang, Jiafu; Chen, Fadi; Lou, Wanghuai; Fang, Weimin; Liu, Zhaolei; Chen, Sumei

2014-07-01

34

A mutation in Arabidopsis that leads to constitutive expression of systemic acquired resistance.  

PubMed

Systemic acquired resistance (SAR) is a nonspecific defense response in plants that is associated with an increase in the endogenous level of salicylic acid (SA) and elevated expression of pathogenesis-related (PR) genes. To identify mutants involved in the regulation of PR genes and the onset of SAR, we transformed Arabidopsis with a reporter gene containing the promoter of a beta-1,3-glucanase-encoding PR gene (BGL2) and the coding region of beta-glucuronidase (GUS). The resulting transgenic line (BGL2-GUS) was mutagenized, and the M2 progeny were scored for constitutive GUS activity. We report the characterization of one mutant, cpr1 (constitutive expressor of PR genes), that was identified in this screen and shown by RNA gel blot analysis also to have elevated expression of the endogenous PR genes BGL2, PR-1, and PR-5. Genetic analyses indicated that the phenotype conferred by cpr1 is caused by a single, recessive nuclear mutation and is suppressed in plants producing a bacterial salicylate hydroxylase, which inactivates SA. Furthermore, biochemical analysis showed that the endogenous level of SA is elevated in the mutant. Finally, the cpr1 plants were found to be resistant to the fungal pathogen Peronospora parasitica NOCO2 and the bacterial pathogen Pseudomonas syringae pv maculicola ES4326, which are virulent in wild-type BGL2-GUS plants. Because the cpr1 mutation is recessive and associated with an elevated endogenous level of SA, we propose that the CPR1 gene product acts upstream of SA as a negative regulator of SAR. PMID:7866028

Bowling, S A; Guo, A; Cao, H; Gordon, A S; Klessig, D F; Dong, X

1994-12-01

35

Premature lethality, hyperactivity, and aberrant phosphorylation in transgenic mice expressing a constitutively active form of Fyn  

PubMed Central

The kinase Fyn, the microtubule-associated protein tau and the peptide amyloid-? (A?) constitute a toxic triad in Alzheimer's disease (AD). Tau's subcellular localization is mainly regulated by phosphorylation whereas Fyn's localization is dictated by palmitoylation targeting it to the plasma membrane in a reversible manner. We have previously shown that tau is required for Fyn to be targeted to the dendritic spine. We had also shown that a truncated form of tau (?tau) that accumulates in the cell soma is capable of trapping Fyn and preventing it from entering the spine. Here we determined that palmitoylation is required for Fyn's membrane and spine localization. We further evaluated the functional consequences of neuronal over-expression of the constitutively active Y531F mutant form of Fyn (FynCA) in transgenic mice. We found that the FynCA transgenic mice displayed a reduced weight, a massively reduced lifespan and a high level of hyperactivity. The lifespan of the FynCA mice was only slightly extended by crossing them with ?tau transgenic mice, possibly reflecting differences in expression patterns of the transgenes and high levels of transgenic FynCA compared to endogenous Fyn. Analysis of synaptosomes revealed that FynCA accumulated at high levels in the spine, resulting in increased levels of the NMDA receptor subunit NR2b phosphorylated at residue Y1472. Tau was strongly phosphorylated at the AT8 epitope S202/T205 as shown by Western blot and immunohistochemistry indicating that an increased tyrosine kinase activity of Fyn has down-stream consequences for serine/threonine-directed phosphorylation. PMID:24860422

Xia, Di; Götz, Jürgen

2014-01-01

36

A mutation in Arabidopsis that leads to constitutive expression of systemic acquired resistance.  

PubMed Central

Systemic acquired resistance (SAR) is a nonspecific defense response in plants that is associated with an increase in the endogenous level of salicylic acid (SA) and elevated expression of pathogenesis-related (PR) genes. To identify mutants involved in the regulation of PR genes and the onset of SAR, we transformed Arabidopsis with a reporter gene containing the promoter of a beta-1,3-glucanase-encoding PR gene (BGL2) and the coding region of beta-glucuronidase (GUS). The resulting transgenic line (BGL2-GUS) was mutagenized, and the M2 progeny were scored for constitutive GUS activity. We report the characterization of one mutant, cpr1 (constitutive expressor of PR genes), that was identified in this screen and shown by RNA gel blot analysis also to have elevated expression of the endogenous PR genes BGL2, PR-1, and PR-5. Genetic analyses indicated that the phenotype conferred by cpr1 is caused by a single, recessive nuclear mutation and is suppressed in plants producing a bacterial salicylate hydroxylase, which inactivates SA. Furthermore, biochemical analysis showed that the endogenous level of SA is elevated in the mutant. Finally, the cpr1 plants were found to be resistant to the fungal pathogen Peronospora parasitica NOCO2 and the bacterial pathogen Pseudomonas syringae pv maculicola ES4326, which are virulent in wild-type BGL2-GUS plants. Because the cpr1 mutation is recessive and associated with an elevated endogenous level of SA, we propose that the CPR1 gene product acts upstream of SA as a negative regulator of SAR. PMID:7866028

Bowling, S A; Guo, A; Cao, H; Gordon, A S; Klessig, D F; Dong, X

1994-01-01

37

Constitutive expression of peroxisome proliferator-activated receptor alpha-regulated genes in dwarf mice.  

PubMed

Defects in growth hormone secretion or signaling in mice are associated with decreased body weights (dwarfism), increased longevity, increased resistance to stress, and decreases in factors that contribute to cardiovascular disease and cancer. Peroxisome proliferators (PP) alter a subset of these changes in wild-type mice through activation of the nuclear receptor family member PP-activated receptor alpha (PPARalpha). We tested the hypothesis that an overlap in the transcriptional programs between untreated dwarf mice and PP-treated wild-type mice underlies these similarities. Using transcript profiling, we observed a statistically significant overlap in the expression of genes differentially regulated in control Snell dwarf mice (Pit-1dw) compared with phenotypically normal heterozygote (+/dw) control mice and those altered by the PP 4-chloro-6-(2,3-xylidino)-2-pyrimidinyl)thioacetic acid (WY-14,643) in +/dw mice. The genes included those involved in beta- and omega-oxidation of fatty acids (Acox1, Cyp4a10, Cyp4a14) and those involved in stress responses (the chaperonin, T-complex protein1epsilon) and cardiovascular disease (fibrinogen). The levels of some of these gene products were also altered in other dwarf mouse models, including Ames, Little, and growth hormone receptor-null mice. The constitutive increases in PPARalpha-regulated genes may be partly caused by increased expression of PPARalpha mRNA and protein as observed in the livers of control Snell dwarf mice. These results indicate that some of the beneficial effects associated with the dwarf phenotype may be caused by constitutive activation of PPARalpha and regulated genes. PMID:15576629

Stauber, Anja J; Brown-Borg, Holly; Liu, Jie; Waalkes, Michael P; Laughter, Ashley; Staben, Rebecca A; Coley, Jaqueline C; Swanson, Cynthia; Voss, Kenneth A; Kopchick, John J; Corton, J Christopher

2005-03-01

38

Constitutive Expression of Mammalian Nitric Oxide Synthase in Tobacco Plants Triggers Disease Resistance to Pathogens  

PubMed Central

Nitric oxide (NO) is known for its role in the activation of plant defense responses. To examine the involvement and mode of action of NO in plant defense responses, we introduced calmodulin-dependent mammalian neuronal nitric oxide synthase (nNOS), which controls the CaMV35S promoter, into wild-type and NahG tobacco plants. Constitutive expression of nNOS led to NO production and triggered spontaneous induction of leaf lesions. Transgenic plants accumulated high amounts of H2O2, with catalase activity lower than that in the wild type. nNOS transgenic plants contained high levels of salicylic acid (SA), and they induced an array of SA-, jasmonic acid (JA)-, and/or ethylene (ET)-related genes. Consequently, NahG co-expression blocked the induction of systemic acquired resistance (SAR)-associated genes in transgenic plants, implying SA is involved in NO-mediated induction of SAR genes. The transgenic plants exhibited enhanced resistance to a spectrum of pathogens, including bacteria, fungi, and viruses. Our results suggest a highly ranked regulatory role for NO in SA-, JA-, and/or ET-dependent pathways that lead to disease resistance. PMID:23124383

Chun, Hyun Jin; Park, Hyeong Cheol; Koo, Sung Cheol; Lee, Ju Huck; Park, Chan Young; Choi, Man Soo; Kang, Chang Ho; Baek, Dongwon; Cheong, Yong Hwa; Yun, Dae-Jin; Chung, Woo Sik; Cho, Moo Je; Kim, Min Chul

2012-01-01

39

Constitutive IRF8 expression inhibits AML by activation of repressed immune response signaling.  

PubMed

Myeloid differentiation is blocked in acute myeloid leukemia (AML), but the molecular mechanisms are not well characterized. Meningioma 1 (MN1) is overexpressed in AML patients and confers resistance to all-trans retinoic acid-induced differentiation. To understand the role of MN1 as a transcriptional regulator in myeloid differentiation, we fused transcriptional activation (VP16) or repression (M33) domains with MN1 and characterized these cells in vivo. Transcriptional activation of MN1 target genes induced myeloproliferative disease with long latency and differentiation potential to mature neutrophils. A large proportion of differentially expressed genes between leukemic MN1 and differentiation-permissive MN1VP16 cells belonged to the immune response pathway like interferon-response factor (Irf) 8 and Ccl9. As MN1 is a cofactor of MEIS1 and retinoic acid receptor alpha (RARA), we compared chromatin occupancy between these genes. Immune response genes that were upregulated in MN1VP16 cells were co-targeted by MN1 and MEIS1, but not RARA, suggesting that myeloid differentiation is blocked through transcriptional repression of shared target genes of MN1 and MEIS1. Constitutive expression of Irf8 or its target gene Ccl9 identified these genes as potent inhibitors of murine and human leukemias in vivo. Our data show that MN1 prevents activation of the immune response pathway, and suggest restoration of IRF8 signaling as therapeutic target in AML. PMID:24957708

Sharma, A; Yun, H; Jyotsana, N; Chaturvedi, A; Schwarzer, A; Yung, E; Lai, C K; Kuchenbauer, F; Argiropoulos, B; Görlich, K; Ganser, A; Humphries, R K; Heuser, M

2015-01-01

40

Pantophysin is a ubiquitously expressed synaptophysin homologue and defines constitutive transport vesicles  

PubMed Central

Certain properties of the highly specialized synaptic transmitter vesicles are shared by constitutively occurring vesicles. We and others have thus identified a cDNA in various nonneuroendocrine cell types of rat and human that is related to synaptophysin, one of the major synaptic vesicle membrane proteins, which we termed pantophysin. Here we characterize the gene structure, mRNA and protein expression, and intracellular distribution of pantophysin. Its mRNA is detected in murine cell types of nonneuroendocrine as well as of neuroendocrine origin. The intron/exon structure of the murine pantophysin gene is identical to that of synaptophysin except for the last intron that is absent in pantophysin. The encoded polypeptide of calculated mol wt 28,926 shares many sequence features with synaptophysin, most notably the four hydrophobic putative transmembrane domains, although the cytoplasmic end domains are completely different. Using antibodies against the unique carboxy terminus pantophysin can be detected by immunofluorescence microscopy in both exocrine and endocrine cells of human pancreas, and in cultured cells, colocalizing with constitutive secretory and endocytotic vesicle markers in nonneuroendocrine cells and with synaptophysin in cDNA-transfected epithelial cells. By immunoelectron microscopy, the majority of pantophysin reactivity is detected at vesicles with a diameter of < 100 nm that have a smooth surface and an electron-translucent interior. Using cell fractionation in combination with immunoisolation, these vesicles are enriched in a light fraction and shown to contain the cellular vSNARE cellubrevin and the ubiquitous SCAMPs in epithelial cells and synaptophysin in neuroendocrine or cDNA-transfected nonneuroendocrine cells and neuroendocrine tissues. Pantophysin is therefore a broadly distributed marker of small cytoplasmic transport vesicles independent of their content. PMID:8707851

1996-01-01

41

Constitutive patterns of gene expression regulated by RNA-binding proteins  

PubMed Central

Background RNA-binding proteins regulate a number of cellular processes, including synthesis, folding, translocation, assembly and clearance of RNAs. Recent studies have reported that an unexpectedly large number of proteins are able to interact with RNA, but the partners of many RNA-binding proteins are still uncharacterized. Results We combined prediction of ribonucleoprotein interactions, based on catRAPID calculations, with analysis of protein and RNA expression profiles from human tissues. We found strong interaction propensities for both positively and negatively correlated expression patterns. Our integration of in silico and ex vivo data unraveled two major types of protein–RNA interactions, with positively correlated patterns related to cell cycle control and negatively correlated patterns related to survival, growth and differentiation. To facilitate the investigation of protein–RNA interactions and expression networks, we developed the catRAPID express web server. Conclusions Our analysis sheds light on the role of RNA-binding proteins in regulating proliferation and differentiation processes, and we provide a data exploration tool to aid future experimental studies. PMID:24401680

2014-01-01

42

Antigen-Specific Monoclonal Antibodies Isolated from B Cells Expressing Constitutively Active STAT5  

PubMed Central

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. PMID:21525985

Scheeren, Ferenc A.; van Geelen, Caroline M. M.; Yasuda, Etsuko; Spits, Hergen; Beaumont, Tim

2011-01-01

43

Constitutive mRNA Expression of Various Glutathione S-Transferase Isoforms in Different Tissues of Mice  

Microsoft Academic Search

Glutathione S-transferase (Gst) enzymes are instrumental in protecting cellular macromolecules against electrophiles and products of oxidative stress. Of interest primarily to pharmacol- ogists and toxicologists is the ability of these enzymes to metabolize cancer chemotherapeutic drugs, insecticides, herbi- cides, and carcinogens. Thus, constitutive expression of Gsts might determine a tissue's ability to handle certain forms of chemical stress. In the

Tamara Raphael Knight; Supratim Choudhuri; Curtis D. Klaassen

2007-01-01

44

Teaching Constitutional Issues with Scripted Trials: Search and Seizure, Freedom of Expression, and the Establishment Clause. Volume 1.  

ERIC Educational Resources Information Center

Intended for use by high school U.S. history and government teachers and students, these scripted trials and related materials are designed to help students develop an understanding about important ideas in the U.S. Constitution. The document focuses on three fundamental issues of the Bill of Rights: search and seizure, freedom of expression, and…

Leming, Robert S.; Vontz, Thomas S.

45

CONSTITUTIVE AND STIMULATED MCP-1, GROA, B, AND Y EXPRESSION IN HUMAN A AIRWAY EPITHELIUM AND BRONCHOALVEOLAR MACROPHAGES  

EPA Science Inventory

Constitutive expression of mRNAs for GROa, GROB, GROY, and MCP-1, belonging to the chemokine family of 8-10 kD cytokines with chemotactic properties for granulocytes and monocytes, has been identified in freshly isolated human nasal and bronchial epithelium, and in bronchoalveola...

46

Electronic Supplemental Material Table 1: Examples of constitutively expressed novel traits of macroevolutionary  

E-print Network

butterflies. Am Nat 112: 999-1015. 7. Otaki JM, 2007. Stress-induced color pattern modifications and evolution Constitutive egg production Periods of ample larval nutrition [1,2] Pontia protodice (checkered white butterfly) Pontia occidentalis (gray- veined white butterfly) Constitutive vernal wing phenotype Unpredictable cold

Abouheif, Ehab

47

[Constitutive expression and purification of Alcaligenes faecalis penicillin G acylase in Escherichia coli].  

PubMed

Considering Alcaligenes faecalis pencillin G acylase(AfPGA), which possesses the attractive characteristics for beta-lactam antibiotics conversions, the gene of PGA was cloned into an expressing vector pKKFPGA. The recombinant plasmid contained multicopy replicon(COLE 1), trc promoter, AfPGA gene, rrnB transcript terminator and ampicillin marker transformed Escherichia coli DH5alpha. As both the recombinant plasmid and the host DH5alpha had no laclq gene, the trc promoter was always active and the AfPGA could be constitutively expressed without IPTG induction in the host DH5alpha. In the shaking flask, the recombinant cell was inoculated into the fermentation medium (tryptone 10g/L, yeast extract 5g/L, MgSO4 x 7 H2O 1g, KH2 PO4 2g/L, K2HPO4 x 3H2O 5g/L, Na2HPO4 x 12H2O 7g/L, (NH4)2SO4 1.2g/L, NH4Cl 0.2 g/L, NaCl 0.1g/L, dextrin 30g/L) and cultured at 28 degrees C for 20h. The production of AfPGA reached 2,590u/L(NIPAB method), with a cell-density-specific activity of more than 300(u/L)/A600, this yield increased 432 fold higher than the native expression of Alcaligenes faecalis . Without ammonium sulphate fractionation and dialysis, the supernatant of crude extract was directly loaded on DEAE-Sepharose CL 6B column equilibrated by phosphate buffer (50mmol/L, pH7.8), and the enzyme fraction was not absorbed on the column but impurities were absorbed. Subsequently the effluent was added ammonium sulphate to 1mol/L and loaded on Butyl-Sepharose CL 4B column equilibrated by 50mmol/L phosphate buffer pH7.8-1mol/L ammonium sulphate. The enzyme was eluted as concentration of ammonium sulphate in phosphate buffer decreased to 0, PGA was eluted. After these two column chromatography, the enzyme was enriched 20 times with a 91% activity recovery. The purified enzyme had a specific activity of 68.6u/mg protein. However, the overproduction of PGA was often limited by translocation and/or periplasmic processing steps, subsequently resulted in intracellular accumulation of various types of PGA precursors and then formed inclusion bodies in the cytoplasm and/or periplasm. In this study, 5% PGA precursors formed as inclusion bodies in the cytoplasm while no inclusion bodies formed in the periplasm. It suggested most PGA precursors were transported to the periplasm and matured to active PGA and also explained why PGA gene was highly expressed in the host DH5alpha. On the other hand, inclusion bodies in the cytoplasm indicated that the maturation of PGA in the host DHSalpha was limited by the translocation step. PMID:15974000

Yang, Zhi-Jian; Cai, Jin; Sun, Jian; Yuan, Zhong-Yi

2004-09-01

48

Paradata for 'ALK1 signalling analysis identifies angiogenesis related genes and reveals disparity between TGF-? and constitutively active receptor induced gene expression'  

NSDL National Science Digital Library

This record contains paradata for the resource 'ALK1 signalling analysis identifies angiogenesis related genes and reveals disparity between TGF-? and constitutively active receptor induced gene expression'

49

Transgenic tobacco plants constitutively expressing Arabidopsis NPR1 show enhanced resistance to root-knot nematode, Meloidogyne incognita  

PubMed Central

In Arabidopsis, non-expressor of pathogenesis related genes-1, NPR1 has been shown to be a positive regulator of the salicylic acid controlled systemic acquired resistance pathway and modulates the cross talk between SA and JA signaling. Transgenic plants expressing AtNPR1 constitutively exhibited resistance against pathogens as well as herbivory. In the present study, tobacco transgenic plants expressing AtNPR1 were studied further for their response to infection by the sedentary endoparasitic root knot nematode, Meloidogyne incognita. Transgenic plants showed enhanced resistance against the root-knot nematode infection. Prominent differences in the shoot and root weights of wild type and transgenic plants were observed post-inoculation with M. incognita. This was associated with a decrease in the number of root galls and egg masses in transgenic plants compared to WT. The transgenic plants also showed constitutive and induced expression of some PR protein genes, when challenged with M. incognita. PMID:21722410

2011-01-01

50

Hypoxia reduces constitutive and TNF-{alpha}-induced expression of monocyte chemoattractant protein-1 in human proximal renal tubular cells  

SciTech Connect

Chronic hypoxia has been reported to be associated with macrophage infiltration in progressive forms of kidney disease. Here, we investigated the regulatory effects of hypoxia on constitutive and TNF-{alpha}-stimulated expression of monocyte chemoattractant protein-1 (MCP-1) in cultured human proximal renal tubular cells (HPTECs). Hypoxia reduced constitutive MCP-1 expression at the mRNA and protein levels in a time-dependent fashion for up to 48 h. Hypoxia also inhibited MCP-1 up-regulation by TNF-{alpha}. Treatment with actinomycin D showed that hypoxic down-regulation of MCP-1 expression resulted mainly from a decrease in the transcription but not the mRNA stability. Immunoblot and immunofluorescence analyses revealed that treatment with hypoxia or an iron chelator, desferrioxamine, induced nuclear accumulation of hypoxia-inducible factor-1{alpha} (HIF-1{alpha}) in HPTECs. Desferrioxamine mimicked hypoxia in the reduction of MCP-1 expression. However, overexpression of a dominant negative form of HIF-1{alpha} did not abolish the hypoxia-induced reduction of MCP-1 expression in HPTECs. These results suggest that hypoxia is an important negative regulator of monocyte chemotaxis to the renal inflamed interstitium, by reducing MCP-1 expression partly via hypoxia-activated signals other than the HIF-1 pathway.

Li Xuan [Division of Nephrology, Department of General Medicine, School of Medicine, Faculty of Medical Sciences, University of Fukui, Fukui (Japan); Kimura, Hideki [Division of Nephrology, Department of General Medicine, School of Medicine, Faculty of Medical Sciences, University of Fukui, Fukui (Japan)]. E-mail: hkimura@fmsrsa.fukui-med.ac.jp; Hirota, Kiichi [Department of Anesthesia, Tazuke Kofukai Medical Research Institute Kitano Hospital, Osaka (Japan); Department of Anesthesia, School of Medicine, Kyoto University, Kyoto (Japan); Sugimoto, Hidehiro [Division of Nephrology, Department of General Medicine, School of Medicine, Faculty of Medical Sciences, University of Fukui, Fukui (Japan); Yoshida, Haruyoshi [Division of Nephrology, Department of General Medicine, School of Medicine, Faculty of Medical Sciences, University of Fukui, Fukui (Japan)

2005-10-07

51

Selection for low dormancy in annual ryegrass (Lolium rigidum) seeds results in high constitutive expression of a glucose-responsive ?-amylase isoform  

PubMed Central

Background and Aims ?-Amylase in grass caryopses (seeds) is usually expressed upon commencement of germination and is rarely seen in dry, mature seeds. A heat-stable ?-amylase activity was unexpectedly selected for expression in dry annual ryegrass (Lolium rigidum) seeds during targeted selection for low primary dormancy. The aim of this study was to characterize this constitutive activity biochemically and determine if its presence conferred insensitivity to the germination inhibitors abscisic acid and benzoxazolinone. Methods ?-Amylase activity in developing, mature and germinating seeds from the selected (low-dormancy) and a field-collected (dormant) population was characterized by native activity PAGE. The response of seed germination and ?-amylase activity to abscisic acid and benzoxazolinone was assessed. Using an alginate affinity matrix, ?-amylase was purified from dry and germinating seeds for analysis of its enzymatic properties. Key Results The constitutive ?-amylase activity appeared late during seed development and was mainly localized in the aleurone; in germinating seeds, this activity was responsive to both glucose and gibberellin. It migrated differently on native PAGE compared with the major activities in germinating seeds of the dormant population, but the enzymatic properties of ?-amylase purified from the low-dormancy and dormant seeds were largely indistinguishable. Seed imbibition on benzoxazolinone had little effect on the low-dormancy seeds but greatly inhibited germination and ?-amylase activity in the dormant population. Conclusions The constitutive ?-amylase activity in annual ryegrass seeds selected for low dormancy is electrophoretically different from that in germinating seeds and its presence confers insensitivity to benzoxazolinone. The concurrent selection of low dormancy and constitutive ?-amylase activity may help to enhance seedling establishment under competitive conditions. PMID:23002268

Goggin, Danica E.; Powles, Stephen B.

2012-01-01

52

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

SciTech Connect

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.

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

1996-05-01

53

The impact of constitutive heterologous expression of a moss Na + transporter on the metabolomes of rice and barley  

Microsoft Academic Search

The metabolic profiles of rice and barley plants constitutively expressing a sodium-pumping ATPase (PpENA1) isolated from the bryophyte Physcomitrella patens were examined using GC-MS. Quantitative real-time PCR (qRT-PCR) was used to determine the mRNA levels of PpENA1 in root and leaf tissues of the transgenic rice and barley lines. PpENA1 mRNA levels were significantly higher in rice lines than in

Andrew Jacobs; Christina Lunde; Antony Bacic; Mark Tester; Ute Roessner

2007-01-01

54

Transactivation of a DR1 PPRE by a human constitutive androstane receptor variant expressed from internal protein translation start sites  

Microsoft Academic Search

Downstream in-frame start codons produce amino- terminal-truncated human constitutive androstane receptor protein isoforms (NCARs). The NCARs are expressed in liver and in vitro cell systems following translation from in-frame methionine AUG start codons at positions 76, 80, 125, 128, 168 and 265 within the full-length CAR mRNA. The resulting CAR proteins lack the N-terminal DNA-binding domain (DBD) of the receptor,

Matthew A. Stoner; Scott S. Auerbach; Stephanie M. Zamule; Stephen C. Strom; Curtis J. Omiecinski

2007-01-01

55

Constitutive activation of the gastrin-releasing peptide receptor expressed by the nonmalignant human colon epithelial cell line NCM460.  

PubMed Central

Gastrin-releasing peptide (GRP) causes multiple effects in humans by activating a specific heptaspanning receptor. Within the gastrointestinal tract, GRP receptors (GRP-R) are not normally expressed by mucosal epithelial cells except for those lining the gastric antrum. In contrast, recent studies have shown that up to 40% of resected colon cancers aberrantly express this receptor. This is important because the GRP-R can cause the proliferation of many, but not all, tissues in which it is expressed. Since GRP and other agonists are not known to exist in the colonic lumen, it has not been clear how or even if GRP-R expression in colon cancer contributes to cell proliferation. To evaluate the functional consequence of GRP-R expression on colonic epithelium, we transfected the recently isolated nonmalignant human colon epithelial cell line NCM460 with the cDNA for this receptor. All NCM460 cell lines expressing varying numbers of GRP-R bound selected agonists and antagonists indistinguishably from receptors expressed by other human tissues. Furthermore GRP-R-expressing transfected cell lines, but not wild-type NCM460 cells, proliferated independently of serum or other growth factors. Further evaluation revealed that GRP-R in these cells tonically stimulated G alpha q/11, resulting in increased phospholipase C activation. Since transfected cells do not secrete GRP, nor is their growth influenced by exposure to receptor-specific antagonists, these data indicate that GRP-R ectopically expressed by NCM460 cells are constitutively active. This report provides the first evidence of mutation-independent heptaspanning receptor constitutive activation resulting in cell proliferation, and identifies a potential mechanism whereby the GRP-R may act as an oncogene in human colon cancer. PMID:9366567

Ferris, H A; Carroll, R E; Rasenick, M M; Benya, R V

1997-01-01

56

High-Level Expression, Purification and Characterization of a Constitutively Active Thromboxane A2 Receptor Polymorphic Variant  

PubMed Central

G protein-coupled receptors (GPCRs) exhibit some level of basal signaling even in the absence of a bound agonist. This basal or constitutive signaling can have important pathophysiological roles. In the past few years, a number of high resolution crystal structures of GPCRs have been reported, including two crystal structures of constitutively active mutants (CAM) of the dim-light receptor, rhodopsin. The structural characterizations of CAMs are impeded by the lack of proper expression systems. The thromboxane A2 receptor (TP) is a GPCR that mediates vasoconstriction and promotes thrombosis in response to the binding of thromboxane. Here, we report on the expression and purification of a genetic variant and CAM in TP, namely A160T, using tetracycline-inducible HEK293S-TetR and HEK293S (GnTIŻ)-TetR cell lines. Expression of the TP and the A160T genes in these mammalian cell lines resulted in a 4-fold increase in expression to a level of 15.8 ±0.3 pmol of receptor/mg of membrane protein. The receptors expressed in the HEK293S (GnTI-)-TetR cell line showed homogeneous glycosylation. The functional yield of the receptors using a single step affinity purification was 45 µg/106 cells. Temperature- dependent secondary structure changes of the purified TP and A160T receptors were characterized using circular dichroism (CD) spectropolarimetry. The CD spectra shows that the loss of activity or thermal sensitivity that was previously observed for the A160T mutant, is not owing to large unfolding of the protein but rather to a more subtle effect. This is the first study to report on the successful high-level expression, purification, and biophysical characterization of a naturally occurring, diffusible ligand activated GPCR CAM. PMID:24086743

Xu, Bing; Chakraborty, Raja; Eilers, Markus; Dakshinamurti, Shyamala; O’Neil, Joe D.; Smith, Steven O.; Bhullar, Rajinder P.; Chelikani, Prashen

2013-01-01

57

Inducible and constitutive expression of an elicitor gene Hrip1 from Alternaria tenuissima enhances stress tolerance in Arabidopsis.  

PubMed

Hrip1 is a novel hypersensitive response-inducing protein secreted by Alternaria tenuissima that activates defense responses and systemic acquired resistance in tobacco. This study investigates the role that Hrip1 plays in responses to abiotic and biotic stress using transgenic Arabidopsis thaliana expressing the Hrip1 gene under the control of the stress-inducible rd29A promoter or constitutive cauliflower mosaic virus 35S promoter. Bioassays showed that inducible Hrip1 expression in rd29A?Hrip1 transgenic lines had a significantly higher effect on plant height, silique length, plant dry weight, seed germination and root length under salt and drought stress compared to expression in 35S?Hrip1 lines and wild type plants. The level of enhancement of resistance to Botrytis cinerea by the 35S?Hrip1 lines was higher than in the rd29A?Hrip1 lines. Moreover, stress-related gene expression in the transgenic Arabidopsis lines was significantly increased by 200 mM NaCl and 200 mM mannitol treatments, and defense genes in the jasmonic acid and ethylene signaling pathway were significantly up-regulated after Botrytis inoculation in the Hrip1 transgenic plants. Furthermore, the activity of some antioxidant enzymes, such as peroxidase and catalase increased after salt and drought stress and Botrytis infection. These results suggested that the Hrip1 protein contributes to abiotic and biotic resistance in transgenic Arabidopsis and may be used as a useful gene for resistance breeding in crops. Although the constitutive expression of Hrip1 is suitable for biotic resistance, inducible Hrip1 expression is more responsive for abiotic resistance. PMID:25120219

Peng, Xue-Cong; Qiu, De-Wen; Zeng, Hong-Mei; Guo, Li-Hua; Yang, Xiu-Fen; Liu, Zheng

2015-02-01

58

Blockade of estrogen-stimulated proliferation by a constitutively-active prolactin receptor having lower expression in invasive ductal carcinoma.  

PubMed

A comprehensive understanding of prolactin's (PRL's) role in breast cancer is complicated by disparate roles for alternatively-spliced PRL receptors (PRLR) and crosstalk between PRL and estrogen signaling. Among PRLRs, the short form 1b (SF1b) inhibits PRL-stimulated cell proliferation. In addition to ligand-dependent PRLRs, constitutively-active varieties, missing the S2 region of the extracellular domain (?S2), naturally occur. Expression analysis of the ?S2 version of SF1b (?S2SF1b) showed higher expression in histologically-normal contiguous tissue versus invasive ductal carcinoma. To determine the function of ?S2SF1b, a T47D breast cancer line with inducible expression was produced. Induction of ?S2SF1b blocked estrogen-stimulated cell proliferation. Unlike intact SF1b, induction of ?S2SF1b had no effect on PRL-mediated activation of Stat5a. However induction inhibited estrogen's stimulatory effects on serine-118 phosphorylation of estrogen receptor ?, serine-473 phosphorylation of Akt, serine-9 phosphorylation of GSK3?, and c-myc expression. In addition, induction of ?S2SF1b increased expression of the cell cycle-inhibiting protein, p21. Thus, increased expression of ?S2SF1b, such as we demonstrate occurs with the selective PRLR modulator, S179D PRL, would create a physiological state in which estrogen-stimulated proliferation was inhibited, but differentiative responses to PRL were maintained. PMID:25527452

Huang, Kuang-Tzu; Tan, Dunyong; Chen, Kuan-Hui E; Walker, Ameae M

2015-03-28

59

Increased constitutive żSMA and Smad2/3 expression in idiopathic pulmonary fibrosis myofibroblasts is K Ca 3.1-dependent.  

PubMed

BackgroundIdiopathic pulmonary fibrosis is a common and invariably fatal disease with limited therapeutic options. Ca2+-activated KCa3.1 potassium channels play a key role in promoting TGFß1 and bFGF-dependent profibrotic responses in human lung myofibroblasts (HLMFs). We hypothesised that KCa3.1 channel-dependent cell processes regulate HLMF żSMA expression via Smad2/3 signalling pathways.MethodsIn this study we have compared the phenotype of HLMFs derived from non-fibrotic healthy control lungs (NFC) with cells derived from IPF lungs. HLMFs grown in vitro were examined for żSMA expression by immunofluorescence (IF), RT-PCR and flow cytommetry. Basal Smad2/3 signalling was examined by RT-PCR, western blot and immunofluorescence. Two specific and distinct KCa3.1 blockers (TRAM-34 200 nM and ICA-17043 [Senicapoc] 100nM) were used to determine their effects on HLMF differentiation and the Smad2/3 signalling pathways.ResultsIPF-derived HLMFs demonstrated increased constitutive expression of both ż-smooth muscle actin (żSMA) and actin stress fibres, indicative of greater myofibroblast differentiation. This was associated with increased constitutive Smad2/3 mRNA and protein expression, and increased Smad2/3 nuclear localisation. The increased Smad2/3 nuclear localisation was inhibited by removing extracellular Ca2+ or blocking KCa3.1 ion channels with selective KCa3.1 blockers (TRAM-34, ICA-17043). This was accompanied by de-differentiation of IPF-derived HLMFs towards a quiescent fibroblast phenotype as demonstrated by reduced żSMA expression and reduced actin stress fibre formation.ConclusionsTaken together, these data suggest that Ca2+- and KCa3.1-dependent processes facilitate żconstitutiveż Smad2/3 signalling in IPF-derived fibroblasts, and thus promote fibroblast to myofibroblast differentiation. Importantly, inhibiting KCa3.1 channels reverses this process. Targeting KCa3.1 may therefore provide a novel and effective approach for the treatment of IPF and there is the potential for the rapid translation of KCa3.1-directed therapy to the clinic. PMID:25476248

Roach, Katy M; Wulff, Heike; Feghali-Bostwick, Carol; Amrani, Yassine; Bradding, Peter

2014-12-01

60

Constitutive expression of high-affinity sulfate transporter (HAST) gene in Indian mustard showed enhanced sulfur uptake and assimilation  

Microsoft Academic Search

Lycopersicon esculantum sulfate transporter gene (LeST 1.1) encodes a high-affinity sulfate transporter (HAST) located in root epidermis. In this\\u000a study, the LeST 1.1 gene was constitutively expressed in Indian mustard (Brassica juncea cv. Pusa Jai Kisan). Transgenic as well as untransformed plants were grown in sulfur-insufficient (25 and 50 ?M) and sulfur-sufficient\\u000a (1,000 ?M) conditions for 30 days. Two-fold increase was noticed in

M. Z. Abdin; M. Akmal; M. Ram; T. Nafis; P. Alam; M. Nadeem; M. A. Khan; A. Ahmad

61

Control of constitutively-expressed developmentally-activated rat hepatic cytochrome P450 genes.  

PubMed

Cytochromes P450 (P450) collectively refer to a superfamily of heme-containing enzymes that use O2 and electrons from NADPH to insert a single atom of oxygen into any one of a large number of substrates. Two general classes of P450s exist; a relatively limited number of P450 forms, expressed in specialized tissues that are associated with pathways of steroidogenesis and a large number of forms responsible for metabolism of foreign compounds. Most of the latter P450s are expressed in liver, the primary site for metabolism of drugs, unusual dietary compounds and environmental pollutants. Numerous forms of P450 are expressed in liver of untreated animals and these are regulated quite differently. Both developmentally-programmed and sex-specific expressions have been observed. In this review, I will summarize recent findings on the mechanisms by which two P450 genes are regulated in livers of developing rats. The CYP2E1 gene is transcriptionally activated within a few hours after birth while the CYP2C6 gene is activated just prior to rats reaching puberty. These genes are under control of two transcription factors, HNF-1 alpha and DBP, respectively, that themselves are developmentally controlled. PMID:1619850

Gonzalez, F J

1992-06-01

62

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

Technology Transfer Automated Retrieval System (TEKTRAN)

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...

63

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

PubMed Central

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. PMID:24765096

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

2014-01-01

64

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

PubMed

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. PMID:24765096

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

2014-01-01

65

International Constitutional Law (ICL)  

NSDL National Science Digital Library

Hosted by the University of Wuerzburg (Germany), International Constitutional Law (ICL) provides English texts of constitutional documents and links to background information on over seventy countries. Documents are cross-referenced for comparison of constitutional provisions. Also featured are links to Constitutional Court sites, a Model Constitutional Code, a section on German Case Law, a comprehensive list of international organizations, and a strong collection of links to constitutional and international law and constitution sites. The material available at ICL is widely applicable to fields such as political science, international relations, or government, and could be very useful for research projects in both secondary and university classrooms.

1997-01-01

66

Constitutive and xenobiotics-induced expression of a novel CYP3A gene from zebrafish larva  

SciTech Connect

In mammals, CYP3A isozymes collectively comprise the largest portion of the liver and small intestinal CYP protein. They are involved in the metabolism of an extensive range of endogenous substrates and xenobiotics and make a significant contribution to the termination of the action of steroid hormones. A full-length cDNA of CYP3A gene, named CYP3A65, was cloned from zebrafish by RT-PCR. The CYP3A65 mRNA was initially transcribed only in the liver and intestine upon hatching of the zebrafish embryos. Like the human CYP3A genes, CYP3A65 transcription in the foregut region was enhanced by treatment of the zebrafish larvae with the steroid dexamethasone and the macrocyclic antibiotic rifampicin. Differing from mammalian CYP3A genes, CYP3A65 transcription was also elicited by 2,3,7,8-tetrachloro-dibenzo-p-dioxin (TCDD) during early larval stages. Repression of AHR2 translation by antisense morpholino oligonucleotides abrogated both of constitutive and TCDD-stimulated CYP3A65 transcription in larval intestine. These findings suggested that the AHR2 signaling pathway plays an essential role in CYP3A65 transcription.

Tseng, H.-P. [Department of Life Science, National Tsing-Hua University, Hsinchu, Taiwan (China); Hseu, Tzong-Hsiung [Department of Life Science, National Tsing-Hua University, Hsinchu, Taiwan (China); Buhler, Donald R. [Department of Molecular Toxicology, Oregon State University, Corvallis, OR 97331 (United States); Wang, W.-D. [Institute of Bioscience and Biotechnology, National Taiwan Ocean University, Keelung, Taiwan (China); Hu, C.-H. [Institute of Bioscience and Biotechnology, National Taiwan Ocean University, Keelung, Taiwan (China)]. E-mail: chhu@mail.ntou.edu.tw

2005-06-15

67

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

PubMed

• 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. PMID:21251001

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

2011-05-01

68

Characterization of dextran-producing Weissella strains isolated from sourdoughs and evidence of constitutive dextransucrase expression.  

PubMed

The study of exopolysaccharide production by heterofermentative sourdough lactic acid bacteria has shown that Weissella strains isolated from sourdoughs produce linear dextrans containing ?-(1?6) glucose residues with few ?-(1?3) linkages from sucrose. In this study, several dextran-producing strains, Weissella cibaria and Weissella confusa, isolated from sourdough, were characterized according to carbohydrate fermentation, repetitive element-PCR fingerprinting using (GTG)(5) primers and glucansucrase activity (soluble or cell-associated). This study reports, for the first time, the characterization of dextransucrase from Weissella strains using sodium dodecyl sulfate-polyacrylamide gel electrophoresis and in situ polymer production (after incubation with sucrose) from enzymatic fractions harvested from both sucrose and glucose culture media. Results demonstrate that dextransucrase activity was mainly soluble and associated with a constitutive 180-kDa protein. In addition, microsequencing of the active dextransucrase from W. cibaria LBAE-K39 allowed the design of specific primers that could detect the presence of glucansucrase encoding genes similar to GTFKg3 of Lactobacillus fermentum Kg3 and to DSRWC of W. cibaria CMU. This study hence indicates that sourdough Weissella strains synthesize original dextransucrase. PMID:20722740

Bounaix, Marie-Sophie; Robert, Hervé; Gabriel, Valérie; Morel, Sandrine; Remaud-Siméon, Magali; Gabriel, Bruno; Fontagné-Faucher, Catherine

2010-10-01

69

Constitutive expression of the barley HvWRKY38 transcription factor enhances drought tolerance in turf and forage grass ( Paspalum notatum Flugge)  

Microsoft Academic Search

WRKY proteins constitute a family of transcription factors involved in many plant processes, including responses to biotic\\u000a and abiotic stress. A constitutive HvWRKY38 expression cassette was introduced into apomictic bahiagrass cultivar ‘Argentine’ by biolistic gene transfer. HvWRKY38 integration and expression was detected in transgenic bahiagrass plants and their apomictic seed progeny by Southern blot,\\u000a PCR and quantitative real-time RT-PCR analysis,

Xi Xiong; Victoria A. James; Hangning Zhang; Fredy Altpeter

2010-01-01

70

A synthetic promoter library for constitutive gene expression in Lactobacillus plantarum  

Microsoft Academic Search

A synthetic promoter library (SPL) for Lactobacillus plantarum has been developed, which generalizes the approach for obtaining synthetic promoters. The consensus sequence, derived from rRNA promoters extracted from the L. plantarum WCFS1 genome, was kept constant, and the non-consensus sequences were randomized. Construction of the SPL was performed in a vector (pSIP409) previously developed for high-level, inducible gene expression in

Ida Rud; Peter Ruhdal Jensen; Kristine Naterstad; Lars Axelsson

2006-01-01

71

Chronic Lung Injury by Constitutive Expression of Activation-Induced Cytidine Deaminase Leads to Focal Mucous Cell Metaplasia and Cancer  

PubMed Central

Activation-induced cytidine deaminase (AID) is an enzyme required for antibody diversification, and it causes DNA mutations and strand breaks. Constitutive AID expression in mice invariably caused lung lesions morphologically similar to human atypical adenomatous hyperplasia (AAH), which can be a precursor of bronchioloalveolar carcinoma. Similar to AAH, mouse AAH-like lesion (MALL) exhibited signs of alveolar differentiation, judging from the expression of alveolar type II (AT2) cell marker surfactant protein C (SP-C). However, electron microscopy indicated that MALL, which possessed certain features of a mucous cell, is distinct from an AAH or AT2 cell. Although MALL developed in all individuals within 30 weeks after birth, lung tumors occurred in only 10%; this suggests that the vast majority of MALLs fail to grow into visible tumors. MALL expressed several recently described markers of lung alveolar regeneration such as p63, keratin 5, keratin 14, leucine-rich repeat containing G protein-coupled receptor 5 (Lgr5), and Lgr6. Increased cell death was observed in the lungs of AID transgenic mice compared with wild-type mice. Based on these observations, we speculate that MALL is a regenerating tissue compensating for cellular loss caused by AID cytotoxicity. AID expression in such regenerating tissue should predispose cells to malignant transformation via its mutagenic activity. PMID:25659078

Kitamura, Jiro; Uemura, Munehiro; Kurozumi, Mafumi; Sonobe, Makoto; Manabe, Toshiaki; Hiai, Hiroshi; Date, Hiroshi; Kinoshita, Kazuo

2015-01-01

72

Effects of sex differences on constitutive nitric oxide synthase expression and activity in response to pressure overload in rats.  

PubMed

Clinical studies have documented sex differences in left ventricular (LV) hypertrophy patterns, but the mechanisms are so far poorly defined. This study aimed to determine whether 1) severe pressure overload altered expression and/or activity of cardiac constitutive nitric oxide synthase (NOS1 and NOS3) and 2) these changes were modulated according to sex. Analyses were performed 0.4-20 wk after thoracic aortic constriction (TAC) in male and female Wistar rats. Male rats with TAC exhibited early signs of cardiac dysfunction, as shown by echocardiographic and LV end-diastolic pressure measurements, whereas females with TAC exhibited higher LV hypertrophy (+96% vs. males at 20 wk; P < 0.05). After TAC, cardiac NOS1 expression was rapidly induced (0.4 wk) and stable afterward in males (P < 0.05 vs. sham groups), whereas it was delayed in females. Accordingly, specific NOS1 activity was increased by 2 wk in male rats with TAC (+122%; P < 0.001 vs. sham groups) and only by 20 wk in females (+220%; P < 0.001 vs. sham groups). NOS1 activity was correlated with NOS1 level. Regarding cardiac NOS3, expression was unaffected by TAC, and the decrease in activity observed at early and late times in male and female rats with TAC, respectively, is shown to be related to NOS3 allosteric regulator caveolin-1 level. The data demonstrated a unique sex-dependent regulation of the constitutive NOSs in response to TAC in rats; such a difference might play a role in the sex-dependent adaptability of the heart in response to pressure overload. PMID:17890423

Loyer, Xavier; Oliviero, Patricia; Damy, Thibaud; Robidel, Estelle; Marotte, Françoise; Heymes, Christophe; Samuel, Jane-Lise

2007-11-01

73

Biallelic MLH1 SNP cDNA expression or constitutional promoter methylation can hide genomic rearrangements causing Lynch syndrome  

Microsoft Academic Search

BackgroundA positive family history, germline mutations in DNA mismatch repair genes, tumours with high microsatellite instability, and loss of mismatch repair protein expression are the hallmarks of hereditary non-polyposis colorectal cancer (Lynch syndrome). However, in ?10–15% of cases of suspected Lynch syndrome, no disease-causing mechanism can be detected.MethodsOligo array analysis was performed to search for genomic imbalances in patients with

Monika Morak; Udo Koehler; Hans Konrad Schackert; Verena Steinke; Brigitte Royer-Pokora; Karsten Schulmann; Matthias Kloor; Wilhelm Höchter; Josef Weingart; Cortina Keiling; Trisari Massdorf; Elke Holinski-Feder

2011-01-01

74

Constitutive expression of an ISGF2/IRF1 transgene leads to interferon-independent activation of interferon-inducible genes and resistance to virus infection.  

PubMed Central

Interferon (IFN)-stimulated gene factor 2 (ISGF2) plays a role in transcription of the beta IFN (IFN-beta) gene and IFN-stimulated genes (ISGs) and may function as a central mediator of cytokine responses. Constitutive ISGF2 transgene expression resulted in substantial resistance to three RNA virus families. This phenotype was not a consequence of IFN production and may have arisen directly through ISG expression. ISGF2 acted generally as a positive transcription factor through binding sites from several genes, in the context of transient cotransfection. Constitutive transcription of the endogenous IFN-beta gene, and several genes that are normally induced by either IFN-alpha or IFN-gamma, or only by IFN-alpha, was elevated in cells that constitutively express an ISGF2 transgene. However, constitutive and virus-induced levels of IFN-beta mRNA were unaffected in such cell lines. Images PMID:1376370

Pine, R

1992-01-01

75

Loss-of-function of Constitutive Expresser of Pathogenesis Related Genes5 affects potassium homeostasis in Arabidopsis thaliana.  

PubMed

Here, we demonstrate that the reduction in leaf K(+) observed in a mutant previously identified in an ionomic screen of fast neutron mutagenized Arabidopsis thaliana is caused by a loss-of-function allele of CPR5, which we name cpr5-3. This observation establishes low leaf K(+) as a new phenotype for loss-of-function alleles of CPR5. We investigate the factors affecting this low leaf K(+) in cpr5 using double mutants defective in salicylic acid (SA) and jasmonic acid (JA) signalling, and by gene expression analysis of various channels and transporters. Reciprocal grafting between cpr5 and Col-0 was used to determine the relative importance of the shoot and root in causing the low leaf K(+) phenotype of cpr5. Our data show that loss-of-function of CPR5 in shoots primarily determines the low leaf K(+) phenotype of cpr5, though the roots also contribute to a lesser degree. The low leaf K(+) phenotype of cpr5 is independent of the elevated SA and JA known to occur in cpr5. In cpr5 expression of genes encoding various Cyclic Nucleotide Gated Channels (CNGCs) are uniquely elevated in leaves. Further, expression of HAK5, encoding the high affinity K(+) uptake transporter, is reduced in roots of cpr5 grown with high or low K(+) supply. We suggest a model in which low leaf K(+) in cpr5 is driven primarily by enhanced shoot-to-root K(+) export caused by a constitutive activation of the expression of various CNGCs. This activation may enhance K(+) efflux, either indirectly via enhanced cytosolic Ca(2+) and/or directly by increased K(+) transport activity. Enhanced shoot-to-root K(+) export may also cause the reduced expression of HAK5 observed in roots of cpr5, leading to a reduction in uptake of K(+). All ionomic data presented is publically available at www.ionomicshub.org. PMID:22046278

Borghi, Monica; Rus, Ana; Salt, David E

2011-01-01

76

Suberoylanilide Hydroxamic Acid Induces Hypersensitivity to Radiation Therapy in Acute Myelogenous Leukemia Cells Expressing Constitutively Active FLT3 Mutants  

PubMed Central

Histone deacetylase inhibitors (HDIs) have shown promise as candidate radiosensitizer for many types of cancers. However, the mechanisms of action are not well understood, and whether they could have clinical impact on radiotherapy for leukemia is unclear. In this study, we demonstrate that suberoylanilide hydroxamic acid (SAHA) can increase radiosensitivity of acute myeloid leukemia (AML) cells through posttranslational modification of Rad51 protein responses and selective inhibition of the homology-directed repair (HDR) pathway. Our data also showed that AML cells with mutant, constitutively active FMS-like tyrosine kinase-3 (FLT3) were more radiation sensitive, caused by compromised non-homologous end joining (NHEJ) repair. Furthermore, SAHA-induced radiosensitization were enhanced in AML cells with expression of these FLT3 mutants. The results of this study suggest that SAHA, a recently approved HDI in clinical trials, may act as a candidate component for novel conditioning regimens to improve efficacy for AML patients undergoing radiotherapy and chemotherapy. PMID:24367670

Chen, Xufeng; Radany, Eric H.; Wong, Patty; Ma, Shenglin; Wu, Kan; Wang, Bing; Wong, Jeffrey Y. C.

2013-01-01

77

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

USGS Publications Warehouse

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 17?-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.

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

2009-01-01

78

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

PubMed

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 17beta-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. PMID:19376234

Robertson, Laura S; Iwanowicz, Luke R; Marranca, Jamie Marie

2009-06-01

79

Metformin Represses Drug-Induced Expression of CYP2B6 by Modulating the Constitutive Androstane Receptor Signaling  

PubMed Central

Metformin is currently the most widely used drug for the treatment of type 2 diabetes. Mechanistically, metformin interacts with many protein kinases and transcription factors that alter the expression of numerous downstream target genes governing lipid metabolism, cell proliferation, and drug metabolism. The constitutive androstane receptor (CAR, NR1i3), a known xenobiotic sensor, has recently been recognized as a novel signaling molecule, in that its activation could be regulated by protein kinases in addition to the traditional ligand binding. We show that metformin could suppress drug-induced expression of CYP2B6 (a typical target gene of CAR) by modulating the phosphorylation status of CAR. In human hepatocytes, metformin robustly suppressed the expression of CYP2B6 induced by both indirect (phenobarbital) and direct CITCO [6-(4-chlorophenyl)imidazo[2,1-b]1,3thiazole-5-carbaldehyde O-(3,4-dichlorobenzyl)oxime] activators of human CAR. Mechanistic investigation revealed that metformin specifically enhanced the phosphorylation of threonine-38 of CAR, which blocks CAR nuclear translocation and activation. Moreover, we showed that phosphorylation of CAR by metformin was primarily an AMP-activated protein kinase– and extracellular signal-regulated kinase 1/2–dependent event. Additional two-hybrid and coimmunoprecipitation assays demonstrated that metformin could also disrupt CITCO-mediated interaction between CAR and the steroid receptor coactivator 1 or the glucocorticoid receptor-interacting protein 1. Our results suggest that metformin is a potent repressor of drug-induced CYP2B6 expression through specific inhibition of human CAR activation. Thus, metformin may affect the metabolism and clearance of drugs that are CYP2B6 substrates. PMID:24252946

Yang, Hui; Garzel, Brandy; Heyward, Scott; Moeller, Timothy; Shapiro, Paul

2014-01-01

80

17beta-estradiol regulates constitutive nitric oxide synthase expression differentially in the myocardium in response to pressure overload.  

PubMed

Estrogens [E(2)] exert direct and indirect effects that can modulate the development of cardiac disease. However, the precise mechanisms that are involved remain undefined. Our objective was to investigate whether E(2) affected the activity and expression of constitutive nitric oxide synthase (NOS) isoforms (NOS3 and NOS1) in cardiac hypertrophy induced by thoracic aortic constriction (TAC). Ovariectomized (Ovx) and nonovariectomized Wistar rats were subjected to TAC. Ovx animals received E(2) or placebo 3 wk after surgery for 11 wk. Afterward cardiac function and degree of left ventricular hypertrophy were assessed by echocardiography. NOS activity and expression were studied by biochemical techniques. TAC led to significant left ventricular hypertrophy (>90%) irrespective of hormonal status. Cardiac performance declined more in TAC+Ovx (-20%, P < 0.015) than in the two other TAC groups [TAC and TAC+Ovx+E(2)]. Total NOS activity decreased significantly in the Ovx groups. In response to TAC, total NOS activity increased whatever the E(2) status. Specific NOS3 activity dramatically decreased in the Ovx groups (-55%, P < 0.009) and was unaltered by TAC. By using coimmunoprecipitation assays, we showed that NOS3/caveolin-1 complexes negatively regulated NOS3 activity as a function of E(2) status. On the other hand, NOS1 expression and activity were markedly increased in hypertrophied myocardium (P < 0.003), irrespective of E(2) status. This study demonstrates a differential regulation of NOS expression and activity in response to pressure overload and E(2) status, the former being mainly involved in the induction of NOS1, whereas the latter regulated NOS3 activity and in turn cardiac function. PMID:17673519

Loyer, Xavier; Damy, Thibaud; Chvojkova, Zuzana; Robidel, Estelle; Marotte, Françoise; Oliviero, Patricia; Heymes, Christophe; Samuel, Jane-Lise

2007-10-01

81

Expression of constitutively active Rab5 uncouples maturation of the Salmonella-containing vacuole from intracellular replication.  

PubMed

The enteric bacterial pathogen Salmonella typhimurium enters and proliferates within both phagocytic and non-phagocytic host cells. Upon entry, the bacteria reside in membrane-bound vacuoles (SCVs) that mature with time, as evidenced by the sequential loss of early endosomal markers, followed by the selective recruitment of a number of lysosomal membrane glycoproteins (LAMPs). This remodelling process renders the SCVs non-fusogenic with lysosomes and is also thought to create a vacuolar environment permissive for replication. We demonstrate that disruption of the endocytic pathway by the expression of a constitutively active form of the small GTPase rab5 (rab5Q79L) significantly altered the biogenesis of the SCVs without inhibiting bacterial replication in HeLa cells. Expression of rab5Q79L caused the retention of early endosomal markers on SCVs and early acquisition of LAMP2, and led to an increase in the kinetics of intracellular replication. We also demonstrate that a significant fraction of LAMP2 in SCVs is derived from the cell surface via endocytosis rather than via the biosynthetic route. Further, in fibroblasts lacking a functional AP3 adaptor complex, in which all newly synthesized LAMP is delivered to the cell surface, recruitment of LAMP to the SCVs remained unaffected. These findings raise the possibility that all the SCV-associated LAMP could be derived by endocytosis from the cell surface. PMID:11437833

Baldeón, M E; Ceresa, B P; Casanova, J E

2001-07-01

82

Molecular characterization of the constitutive expression of the plasma platelet-activating factor acetylhydrolase gene in macrophages.  

PubMed Central

Plasma platelet-activating factor acetylhydrolase (PAF-AH) is a phospholipase that inactivates platelet-activating factor (PAF) and PAF-like lipids to generate products with little or no biological activity. The levels of circulating PAF-AH correlate with several disease syndromes. We previously reported that mediators of inflammation regulate the expression of the human PAF-AH gene at the transcriptional level. In the present paper, we characterize the constitutive expression of plasma PAF-AH using the mouse gene as a model system, and we report comparative results obtained using human and mouse promoter constructs. We first cloned, sequenced and analysed the promoter region of the murine plasma PAF-AH (mPAF-AH) gene and found that this gene lacks a canonical TATA box. We demonstrated that the cis -elements required for basal transcription are localized within the -316 to -68 bp region. In vitro band-shift and supershift assays showed that Sp1 and Sp3 transcription factors from RAW264.7 and J774A.1 macrophage nuclear extracts bound strongly to a distal GC-rich site within -278/-243 [specificity protein (Sp-A)] and to a proximal TC-rich motif within -150/-114 (Sp-B). In addition, we observed weak binding to a GA-rich site within -110/-82 (Sp-C). The regions containing Sp-B and Sp-C are highly conserved between the human and mouse genes. Forced expression of Sp1 or Sp3 in Sp-lacking Drosophila SL2 cells induced markedly the activity of the exogenous mPAF-AH promoter in a dose-dependent manner, and this induction was dependent on the presence of intact Sp-A and Sp-B. Interestingly, we found that the Sp1- and Sp3-associated DNA-binding activities increased during the maturation of primary human monocytes into macrophages in cell culture. These results demonstrate that Sp1 and Sp3 are key factors that contribute to the basal, constitutive transcription of the plasma PAF-AH gene in macrophages. PMID:12854969

Wu, Xiaoqing; McIntyre, Thomas M; Zimmerman, Guy A; Prescott, Stephen M; Stafforini, Diana M

2003-01-01

83

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

PubMed Central

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

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

2011-01-01

84

Ectopic expression of the phosphomimic mutant version of Arabidopsis response regulator 1 promotes a constitutive cytokinin response phenotype  

PubMed Central

Background Cytokinins control numerous plant developmental processes, including meristem formation and activity, nutrient distribution, senescence timing and responses to both the abiotic and biotic environments. Cytokinin signaling leads to the activation of type-B response regulators (RRBs), Myb-like transcription factors that are activated by the phosphorylation of a conserved aspartate residue in their response receiver domain. Consistent with this, overexpression of RRBs does not substantially alter plant development, but instead leads to cytokinin hypersensitivity. Results Here we present comparative analysis of plants overexpressing Arabidopsis RRB 1 (ARR1) or a phosphomimic ARR1D94E mutant in which the conserved aspartate-94 (D94) is replaced by the phosphomimic residue glutamate (E). The D94E substitution causes a 100-fold increase in response activation and instigates developmental and physiological changes that characterize wild-type plants treated with cytokinins or transgenic plants with increased cytokinin content. Conclusion The current model of cytokinin signaling emphasizes the essential role of conserved aspartate residue phosphorylation of RRBs in promoting cytokinin responses. Our comparative analyses of developmental and physiological traits of ARR1 and ARR1D94E overexpressing plants revealed that the ARR1D94E protein is indeed a constitutive and wide-spectrum cytokinin response activator. PMID:24423196

2014-01-01

85

Ortho-aminoazotoluene activates mouse constitutive androstane receptor (mCAR) and increases expression of mCAR target genes  

SciTech Connect

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{sup -/-}) 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{sup -/-} 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. - Highlights: > The azo dye and mouse carcinogen OAT is a very effective mCAR activator. > OAT increases mCAR transactivation in a dose-dependent manner. > OAT CAR-dependently increases the expression of a specific subset of CAR target genes. > OAT induces an unexpectedly deferred, but CAR-dependent hepatocyte proliferation.

Smetanina, Mariya A., E-mail: maria.smetanina@gmail.com [Department of Molecular and Cellular Biology, Baylor College of Medicine, 1 Baylor Plaza, Houston, TX 77030 (United States); Laboratory of Gene Expression Control, Institute of Cytology and Genetics of the Siberian Branch of the Russian Academy of Sciences, prospekt Lavrentyeva 10, Novosibirsk 630090 (Russian Federation); Group of Pharmacogenomics, Institute of Chemical Biology and Fundamental Medicine of the Siberian Branch of the Russian Academy of Sciences, prospekt Lavrentyeva 8, Novosibirsk 630090 (Russian Federation); Pakharukova, Mariya Y. [Laboratory of Gene Expression Control, Institute of Cytology and Genetics of the Siberian Branch of the Russian Academy of Sciences, prospekt Lavrentyeva 10, Novosibirsk 630090 (Russian Federation); Kurinna, Svitlana M. [Department of Biochemistry and Molecular Biology, Unit 1000, University of Texas M. D. Anderson Cancer Center, 1515 Holcombe Blvd., Houston, TX 77030 (United States); Dong, Bingning; Hernandez, Juan P.; Moore, David D. [Department of Molecular and Cellular Biology, Baylor College of Medicine, 1 Baylor Plaza, Houston, TX 77030 (United States); Merkulova, Tatyana I. [Laboratory of Gene Expression Control, Institute of Cytology and Genetics of the Siberian Branch of the Russian Academy of Sciences, prospekt Lavrentyeva 10, Novosibirsk 630090 (Russian Federation)

2011-08-15

86

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

PubMed

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. PMID:23899775

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

2013-09-01

87

Transactivation of a DR-1 PPRE by a human constitutive androstane receptor variant expressed from internal protein translation start sites  

PubMed Central

Downstream in-frame start codons produce amino-terminal-truncated human constitutive androstane receptor protein isoforms (?NCARs). The ?NCARs are expressed in liver and in vitro cell systems following translation from in-frame methionine AUG start codons at positions 76, 80, 125, 128, 168 and 265 within the full-length CAR mRNA. The resulting CAR proteins lack the N-terminal DNA-binding domain (DBD) of the receptor, yielding ?NCAR variants with unique biological function. Although the ?NCARs maintain full retinoid X receptor alpha (RXR?) heterodimerization capacity, the ?NCARs are inactive on classical CAR-inducible direct repeat (DR)-4 elements, yet efficiently transactivate a DR-1 element derived from the endogenous PPAR-inducible acyl-CoA oxidase gene promoter. RXR? heterodimerization with CAR1, CAR76 and CAR80 isoforms is necessary for the DR-1 PPRE activation, a function that exhibits absolute dependence on both the respective RXR? DBD and CAR activation (AF)-2 domains, but not the AF-1 or AF-2 domain of RXR?, nor CAR's DBD. A new model of CAR DBD-independent transactivation is proposed, such that in the context of a DR-1 peroxisome proliferator-activated response element, only the RXR? portion of the CAR-RXR? heterodimer binds directly to DNA, with the AF-2 domain of tethered CAR mediating transcriptional activation of the receptor complex. PMID:17355985

Stoner, Matthew A.; Auerbach, Scott S.; Zamule, Stephanie M.; Strom, Stephen C.; Omiecinski, Curtis J.

2007-01-01

88

Characterization and constitutive expression of a novel endo-1,4-?-D-xylanohydrolase from Aspergillus niger in Pichia pastoris.  

PubMed

A putative endo-1,4-?-D-xylanohydrolase gene xyl10 from Aspergillus niger, encoding a 308-residue mature xylanase belonging to glycosyl hydrolase family 10, was constitutively expressed in Pichia pastoris. The recombinant Xyl10 exhibited optimal activity at pH 5.0 and 60 °C with more than 50 % of the maximum activity from 40 to 70 °C. It retained more than 90 % of the original activity after incubation at 60 °C (pH 5.0) for 30 min and more than 74 % after incubation at pH 3.0-13.0 for 2 h (25 °C). The specific activity, K m and V max values for purified Xyl10 were, respectively, 3.2 × 10(3) U mg(-1), 3.6 mg ml(-1) and 5.4 × 10(3) ?mol min(-1 )mg(-1) towards beechwood xylan. The enzyme degraded xylan to a series of xylooligosaccharides and xylose. The recombinant enzyme with these properties has the potential for various industrial applications. PMID:23690032

Zheng, Jia; Guo, Ning; Wu, Lishuang; Tian, Jian; Zhou, Hongbo

2013-09-01

89

Transient Cardiac Expression of Constitutively Active Galpha q Leads to Hypertrophy and Dilated Cardiomyopathy by Calcineurin-Dependent and Independent Pathways  

Microsoft Academic Search

Cardiac hypertrophy and dilatation can result from stimulation of signal transduction pathways mediated by heterotrimeric G proteins, especially Gq, whose alpha subunit activates phospholipase Cbeta (PLCbeta ). We now report that transient, modest expression of a hemagglutinin (HA) epitope-tagged, constitutively active mutant of the Gqalpha subunit (HAalpha q*) in hearts of transgenic mice is sufficient to induce cardiac hypertrophy and

U. Mende; A. Kagen; A. Cohen; J. Aramburu; F. J. Schoen; E. J. Neer

1998-01-01

90

Constitutive Expression of the Maize Genes B1 and C1 in Transgenic Hi II Maize Results in Differential Tissue Pigmentation and Generates Resistance to Helicoverpa zea  

Technology Transfer Automated Retrieval System (TEKTRAN)

Anthocyanin biosynthesis in maize protects tissues from biotic and abiotic stresses. Constitutive expression of the maize B1 and C1 genes, which induces anthocyanin biosynthesis, resulted in transgenic plants with varied phenotypes. Some colored leaves were substantially resistant to thrips damage...

91

Constitutional Legitimacy  

Microsoft Academic Search

The problem of constitutional legitimacy is to establish why anyone should obey the command of a constitutionally-valid law. A lawmaking system is legitimate if there is a prima facie duty to obey the laws it makes. Neither \\

Randy E. Barnett

2003-01-01

92

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

PubMed

Enzymes for use in the sugar industry are preferred to be thermotolerant. In this study, a synthetic codon-optimized gene encoding a highly thermostable ?-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 ?-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. PMID:22821437

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

2013-02-01

93

Constitutional Issues and Iowa.  

ERIC Educational Resources Information Center

Important constitutional issues are presented in a manner appropriate for use in the classroom. Case studies and events from the history of Iowa are used to illuminate the Constitution and Bill of Rights. Freedom of expression and students' rights are discussed in "The Black Armband Case"; free exercise of religion as won by the Iowa's Amish is…

Gore, Deborah, Ed.

1987-01-01

94

Strain background influences neurotoxicity and behavioral abnormalities in mice expressing the tetracycline transactivator  

PubMed Central

The tet-off system has been widely used to create transgenic models of neurological disorders including Alzheimer’s, Parkinson’s, Huntington’s, and prion disease. The utility of this system lies in the assumption that the tetracycline transactivator (TTA) acts as an inert control element and does not contribute to phenotypes under study. Here we report that neuronal expression of TTA can affect hippocampal cytoarchitecture and behavior in a strain-dependent manner. While studying neurodegeneration in two tet-off Alzheimer’s disease models, we unexpectedly discovered neuronal loss within the dentate gyrus of single transgenic TTA controls. Granule neurons appeared most sensitive to TTA exposure during postnatal development, and doxycycline treatment during this period was neuroprotective. TTA-induced degeneration could be rescued by moving the transgene onto a congenic C57BL/6J background, and recurred on re-introduction of either CBA or C3H/He backgrounds. Quantitative trait analysis of B6C3 F2 TTA mice identified a region on Chromosome 14 that contains a major modifier of the neurodegenerative phenotype. Although B6 mice were resistant to degeneration, they were not ideal for cognitive testing. F1 offspring of TTA C57BL/6J and 129X1/SvJ, FVB/NJ, or DBA/1J showed improved spatial learning, but TTA expression caused subtle differences in contextual fear conditioning on two of these backgrounds indicating that strain and genotype can interact independently under different behavioral settings. All model systems have limitations that should be recognized and mitigated where possible; our findings stress the importance of mapping the effects caused by TTA alone when working with tet-off models. PMID:22855807

Han, Harry J.; Allen, Carolyn C.; Buchovecky, Christie M.; Yetman, Michael J.; Born, Heather A.; Marin, Miguel A.; Rodgers, Shaefali P.; Song, Bryan J.; Lu, Hui-Chen; Justice, Monica J.; Probst, Frank J.; Jankowsky, Joanna L.

2012-01-01

95

Destabilizing Protein Polymorphisms in the Genetic Background Direct Phenotypic Expression of Mutant SOD1 Toxicity  

PubMed Central

Genetic background exerts a strong modulatory effect on the toxicity of aggregation-prone proteins in conformational diseases. In addition to influencing the misfolding and aggregation behavior of the mutant proteins, polymorphisms in putative modifier genes may affect the molecular processes leading to the disease phenotype. Mutations in SOD1 in a subset of familial amyotrophic lateral sclerosis (ALS) cases confer dominant but clinically variable toxicity, thought to be mediated by misfolding and aggregation of mutant SOD1 protein. While the mechanism of toxicity remains unknown, both the nature of the SOD1 mutation and the genetic background in which it is expressed appear important. To address this, we established a Caenorhabditis elegans model to systematically examine the aggregation behavior and genetic interactions of mutant forms of SOD1. Expression of three structurally distinct SOD1 mutants in C. elegans muscle cells resulted in the appearance of heterogeneous populations of aggregates and was associated with only mild cellular dysfunction. However, introduction of destabilizing temperature-sensitive mutations into the genetic background strongly enhanced the toxicity of SOD1 mutants, resulting in exposure of several deleterious phenotypes at permissive conditions in a manner dependent on the specific SOD1 mutation. The nature of the observed phenotype was dependent on the temperature-sensitive mutation present, while its penetrance reflected the specific combination of temperature-sensitive and SOD1 mutations. Thus, the specific toxic phenotypes of conformational disease may not be simply due to misfolding/aggregation toxicity of the causative mutant proteins, but may be defined by their genetic interactions with cellular pathways harboring mildly destabilizing missense alleles. PMID:19266020

Gidalevitz, Tali; Krupinski, Thomas; Garcia, Susana; Morimoto, Richard I.

2009-01-01

96

Immunologic Self-Tolerance Maintained by CD25 1 CD4 1 Regulatory T Cells Constitutively Expressing Cytotoxic T Lymphocyte-associated Antigen 4  

Microsoft Academic Search

This report shows that cytotoxic T lymphocyte-associated antigen 4 (CTLA-4) plays a key role in T cell-mediated dominant immunologic self-tolerance. In vivo blockade of CTLA-4 for a limited period in normal mice leads to spontaneous development of chronic organ-specific au- toimmune diseases, which are immunopathologically similar to human counterparts. In normal naive mice, CTLA-4 is constitutively expressed on CD25 1

Takeshi Takahashi; Tomoyuki Tagami; Sayuri Yamazaki; Toshimitsu Uede; Jun Shimizu; Noriko Sakaguchi; Tak W. Mak; Shimon Sakaguchi

97

[Expression of the symbiotic nod4 pea gene against various genotypic backgrounds].  

PubMed

Experiments on the effect of genotypic environment on the expression of the nod4 gene, responsible for supernodulation in pea, were performed. The genotypic background was found to affect the manifestation of both major symbiosis-related traits (number of nitrogen-fixing bacterial nodules and nitrogenase activity) and productivity-related traits (stem height, seed number, and seed weight), which form the pleiotropic complex of the mutant gene. Using recurrent selection, we developed supernodulating lines significantly exceeding the original mutant line and studied them up to generations F5-F6. Of special importance is the fact that these lines showed high levels of nodulation and nitrogen fixation. The results are discussed from the viewpoint of breeding. PMID:15977818

Sidorova, K K; Shumny?, V K; Glianenko, M N; Mishchenko, T M; Vlasova, E Iu; Gaeva, T A

2005-05-01

98

Constitutive expression of the embryonic stem cell marker OCT4 in bovine somatic donor cells influences blastocysts rate and quality after nucleus transfer.  

PubMed

Nuclear transfer (NT) is associated with epigenetic reprogramming of donor cells. Expression of certain genes in these cells might facilitate their expression in the NT embryo. This research was aimed to investigate the effect of constitutive expression of OCT4 in bovine somatic cells used for NT on the developmental potential of derived cloned embryos as well as in the expression of pluripotency markers in the Day-7 resulting embryos. Cloned blastocysts were generated from five cell lines that expressed OCT4. Pools of blastocysts were screened to detect OCT4, SOX2, and NANOG by qPCR. In vitro-fertilized time-matched blastocysts were used as controls. The development potential was assessed on the basis of blastocysts rate; grading and total cell counts at Day 7. OCT4 expression in the cell lines positively correlates with blastocysts rate (r = 0.92; p = 0.02), number of grade I blastocysts (r = 0.96; p = 0.01), and total cell number (r = 0.98; p = 0.002). The high expression of OCT4 in the cell line did not improve the final outcome of cloning. Somatic expression of OCT4 lead to increased expression of OCT4 and SOX2 in cloned grade I blastocysts; however, there was a bigger variability in OCT4 and SOX2 (p = 0.03; p = 0.02) expression in the embryos generated from cells expressing highest levels of OCT4. Probably the higher variability in OCT4 expression in cloned embryos is due to incorrect reprogramming and incapability of the oocyte to correct for higher OCT4 levels. For that reason, we concluded that OCT4 expression in somatic cells is not a good prognosis marker for selecting cell lines. PMID:23846396

Rodríguez-Alvarez, Lleretny; Manriquez, Jose; Velasquez, Alejandra; Castro, Fidel Ovidio

2013-10-01

99

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

Microsoft Academic Search

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

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

2006-01-01

100

Constitution Daily  

NSDL National Science Digital Library

Everyone could use a bit of the Constitution added to their daily lives, and this website more than delivers on its promise to deliver "smart conversation about the Constitution". Constitution Daily is an experimental blog edited by the National Constitution Center (NCC) in Philadelphia, and commentary here can include conversations about student privacy rights, the Second Amendment, and the activities of Congress. Visitors can click on the "Issues" section to dive into topic areas that include civility and privacy. After looking at each topic area, visitors can look at an interactive timeline that arranges comments, posts, and discussion on the subject. The site also contains some nice polls, and information about upcoming events at the NCC.

101

Constitutive and salt-inducible expression of SlBADH gene in transgenic tomato (Solanum lycopersicum L. cv. Micro-Tom) enhances salt tolerance.  

PubMed

To improve the stress tolerance of crops, many genes, including transcription factors, have been expressed in transgenic plants using either constitutive or stress-inducible promoters. However, transgenic plants that show strong constitutive expression of transcription factors often suffer from many undesirable phenotypes, such as stunted growth and reduced yield. In the present study, the betaine aldehyde dehydrogenase (BADH) gene, cloned from Suaeda liaotungensis and, controlled by the Cauliflower mosaic virus (CaMV) 35S promoter or stress-inducible promoter of BADH (P5: -300 to +62 bp), was transformed into tomato (Solanum lycopersicum). The transformants with single copy of SlBADH were determined by real time PCR. Expression of SlBADH in the P5:BADH transgenic plants exhibited salt induced and was higher than that in CaMV35S:BADH under salt stress. The SlBADH enhanced salt tolerance of P5:BADH and CaMV35S:BADH transformants. And SlBADH in P5:BADH plants did not affect the growth of transformants. Consequently, we conclude that the P5 promoter can drive increased expression of SlBADH in transgenic tomato under salt stress and increase salt tolerance without affecting plant growth. PMID:23402752

Wang, Jing-yu; Lai, Lu-di; Tong, Shao-ming; Li, Qiu-li

2013-03-01

102

Constitutive expression of a complement-like protein in Toll and JAK gain-of-function mutants of Drosophila  

Microsoft Academic Search

We show that Drosophila expresses four genes encoding proteins with significant similarities with the thiolester-containing proteins of the complement C3\\/2-macroglobulin superfamily. The genes are transcribed at a low level during all stages of development, and their expression is markedly up-regulated after an immune challenge. For one of these genes, which is predominantly expressed in the larval fat body, we observe

Marie Lagueux; Emmanuel Perrodou; Elena A. Levashina; Maria Capovilla; Jules A. Hoffmann

2000-01-01

103

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

PubMed Central

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 F2? 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.

Klaus, Günther

2015-01-01

104

Transcriptional downregulation of agr expression in Staphylococcus aureus during growth in human serum can be overcome by constitutively active mutant forms of the sensor kinase AgrC.  

PubMed

The temporal and cell density-dependent regulation of expression of virtually all the Staphylococcus aureus virulon is under the control of the agr (accessory gene regulatory) operon. The expression of the agr operon is subject to transcriptional regulation by the AgrA/C two-component response regulator/sensor kinase pair. During bacteraemia, a frequent syndrome caused by methicillin-resistant S. aureus (MRSA), the transcriptional downregulation of agr expression has been attributed to the sequestration of the quorum-signalling molecule auto-inducing peptide (AIP) by the human serum component apolipoprotein B as part of an innate immune response to infection. However, it is not known whether transcriptional downregulation of agr expression during growth in human serum is additionally subjected to regulation by transcription regulatory proteins that either directly or indirectly affect transcription from the agr operon promoters. Here, using chromosomal fluorescence reporters of agr expression in S. aureus, we show that the transcriptional downregulation of agr expression in human serum can be overcome using constitutive active mutant forms of AgrC. Therefore, it seems that the sequestration of the AIP is likely to be the only mechanism by which the host innate immune response limits agr expression at the transcriptional level to maintain the host-pathogen balance towards a noninvasive outcome. PMID:24164684

James, Ellen H; Edwards, Andrew M; Wigneshweraraj, Sivaramesh

2013-12-01

105

Transcriptional downregulation of agr expression in Staphylococcus aureus during growth in human serum can be overcome by constitutively active mutant forms of the sensor kinase AgrC  

PubMed Central

The temporal and cell density-dependent regulation of expression of virtually all the Staphylococcus aureus virulon is under the control of the agr (accessory gene regulatory) operon. The expression of the agr operon is subject to transcriptional regulation by the AgrA/C two-component response regulator/sensor kinase pair. During bacteraemia, a frequent syndrome caused by methicillin-resistant S. aureus (MRSA), the transcriptional downregulation of agr expression has been attributed to the sequestration of the quorum-signalling molecule auto-inducing peptide (AIP) by the human serum component apolipoprotein B as part of an innate immune response to infection. However, it is not known whether transcriptional downregulation of agr expression during growth in human serum is additionally subjected to regulation by transcription regulatory proteins that either directly or indirectly affect transcription from the agr operon promoters. Here, using chromosomal fluorescence reporters of agr expression in S. aureus, we show that the transcriptional downregulation of agr expression in human serum can be overcome using constitutive active mutant forms of AgrC. Therefore, it seems that the sequestration of the AIP is likely to be the only mechanism by which the host innate immune response limits agr expression at the transcriptional level to maintain the host–pathogen balance towards a noninvasive outcome. PMID:24164684

James, Ellen H; Edwards, Andrew M; Wigneshweraraj, Sivaramesh

2013-01-01

106

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

PubMed

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. PMID:18440928

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

2008-01-01

107

Constitutive expression of CD26/dipeptidylpeptidase IV on peripheral blood B lymphocytes of patients with B chronic lymphocytic leukaemia  

PubMed Central

We have investigated the expression of the ectoenzyme dipeptidylpeptidase IV (DPP IV)/CD26 on lymphocytes obtained from patients with B chronic lymphocytic leukaemia (B-CLL) and compared it with healthy subjects. Using two-colour immunofluorescence analysis with CD26 and CD20 or CD23 monoclonal antibodies, CD26 was found undetectable on peripheral resting B-cells (CD20+ CD23?) from normal donors whereas it was expressed on B-cells activated in vitro with interleukin (IL)-4 and Staphylococcus aureus strain cowan I (CD20+ CD23+). The expression of CD26 on leukaemic B-cells (CD20+ CD23+) was clearly induced in 22 out of 25 patients examined. Consequently, induced levels of CD26 cell surface expression on either normal activated and malignant B-cells coincided with the enhancement of DPP IV activity detected on the surface of these cells. Reverse transcription polymerase chain reaction analyses showed that the transcript levels of the CD26 gene was higher in normal activated B-cells and B-CLL cells than in resting B-cells, suggesting that CD26 was expressed at the level of transcriptional activation. These observations provide evidence of the abnormal expression of DPPIV/CD26 in B-CLL which, therefore, may be considered as a novel marker for B-CLL. Further investigation in relation to CD26 expression and other B malignancies needs to be defined. © 1999 Cancer Research Campaign PMID:10098734

Bauvois, B; Meester, I De; Dumont, J; Rouillard, D; Zhao, H X; Bosmans, E

1999-01-01

108

High expression of Janus kinase 2 in background normal liver tissue of resected hepatocellular carcinoma is associated with worse prognosis.  

PubMed

When assessing hepatocellular carcinoma (HCC), it is important to examine prognostic factors in the background normal liver tissue and consider malignant aspects of the primary lesion. Candidate genes were extracted from the background normal liver samples via multiarray analysis. Control samples, termed supernormal (SN) liver, were obtained from 11 cases of metastatic liver cancer. Corresponding normal (CN) liver tissue was surgically obtained from a typical HCC patient with chronic hepatitis C background for comparison. Expression profile and methylation array demonstrated that Janus kinase 2 (JAK2) gene expression was increased by 2.378?fold in the CN tissue. Methylation array reported a lower value for CN (0.125) than SN tissues (0.748). We then investigated JAK2 expression by real-time quantitative reverse transcription-polymerase chain reaction in 100 consecutive resected HCC cases. The average expression level of JAK2 (normalized to GAPDH) was significantly lower in CN (9.24±6.43, n=100) than in SN (35.21±21.38, n=11) tissues (P<0.001). As such a result was contrary to our expectation, the case used for array analysis seemed to be a rare incidence. One hundred HCC cases were subsequently divided into two groups based on JAK2 expression in the adjacent normal tissue: one consisting of the upper 70% of cases (n=70) and the other of the remaining 30% (n=30). Higher JAK2 expression in the adjacent tissue demonstrated significant correlation with worse survival (P=0.022). Furthermore, multivariate analysis identified higher JAK2 expression in the background normal liver tissue of HCC as an independent prognostic factor (P=0.032). Our findings suggest that higher JAK2 expression in the background normal liver tissue of HCC may be a good prognostic biomarker for resected HCC. PMID:25420511

Sonohara, Fuminori; Nomoto, Shuji; Inokawa, Yoshikuni; Hishida, Mitsuhiro; Takano, Nao; Kanda, Mitsuro; Nishikawa, Yoko; Fujii, Tsutomu; Koike, Masahiko; Sugimoto, Hiroyuki; Kodera, Yasuhiro

2015-02-01

109

Constitutive expression and phenobarbital modulation of drug metabolizing enzymes and related nuclear receptors in cattle liver and extra-hepatic tissues.  

PubMed

In humans and rodents, phenobarbital (PB) induces hepatic and extra-hepatic drug metabolizing enzymes (DMEs) through the activation of specific nuclear receptors (NRs). In contrast, few data about PB transcriptional effects in veterinary species are available. The constitutive expression and modulation of PB-responsive NR and DME genes, following an oral PB challenge, were investigated in cattle liver and extra-hepatic tissues (duodenum, kidney, lung, testis, adrenal and muscle). Likewise to humans and rodents, target genes were expressed to a lower extent compared to the liver with few exceptions. Phenobarbital significantly affected hepatic CYP2B22, 2C31, 2C87, 3A and UDP-glucuronosyltransferase 1A1-like, glutathione S-transferase A1-like and sulfotransferase 1A1-like (SULT1A1-like) mRNAs and apoprotein amounts; in extra-hepatic tissues, only duodenum showed a significant down-regulation of SULT1A1-like gene and apoprotein. Nuclear receptor mRNAs were never affected by PB. Presented data are the first evidence about the constitutive expression of foremost DME and NR genes in cattle extra-hepatic tissues, and the data obtained following a PB challenge are suggestive of species-differences in drug metabolism; altogether, these information are of value for the extrapolation of pharmacotoxicological data among species, the characterization of drug-drug interactions as well as the animal and consumer's risk caused by harmful residues formation. PMID:22694178

Zancanella, Vanessa; Giantin, Mery; Lopparelli, Rosa Maria; Nebbia, Carlo; Dacasto, Mauro

2012-11-01

110

Constitution Annotated  

NSDL National Science Digital Library

What exactly does the United States Constitution mean? It is subject, of course, to ongoing meditation, debate, argument, and interpretation. This useful site provides one lens of interpretation of the Constitution, based primarily on Supreme Court case law and legal analysis. Visitors can look over each section of the Constitution individually, including all of the articles and amendments. Additionally, the site contains a complete table of Supreme Court cases, along with those cases that have been overruled as well as proposed amendments that have not been ratified. Additionally, the site contains a gallery of significant primary source documents from American history. Interested users should not forget to download the helpful app that is also included here.

2013-06-26

111

The Constitution: Perspectives on Contemporary American Democracy.  

ERIC Educational Resources Information Center

Four articles expressing the views of nine prominent United States citizens about the Constitution provide a context for reflecting on the meaning of the Constitution in present-day America. In "Why Has the Constitution Endured So Long?" Don Edwards, chairman of the House Civil and Constitutional Rights Subcommittee, discusses why the Constitution

Close Up Foundation, Arlington, VA.

112

Constitutional Conservatism  

ERIC Educational Resources Information Center

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…

Berkowitz, Peter

2009-01-01

113

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

PubMed

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. PMID:11449302

Monesi, N; Sousa, J F; Paçó-Larson, M L

2001-07-01

114

Expression of a constitutive form of calcium/calmodulin dependent protein kinase II leads to arrest of the cell cycle in G2.  

PubMed Central

Calcium/calmodulin dependent protein kinase II (CaMKII) is a multifunctional serine/threonine protein kinase. We have created a calcium/calmodulin independent form of this enzyme by truncation. Expression of this enzyme fragment in a rabbit reticulocyte lysate yields a constitutive enzyme with specific activity similar to the activated native enzyme. We have established mammalian cell lines that transiently express this constitutive enzyme using the glucocorticoid-inducible mouse mammary tumor virus long terminal repeat. The transient increase in kinase activity results in a complete cessation of cell cycle progression. This block develops as a consequence of a specific arrest of the cell cycle in G2. During the block, increases in histone H1 kinase activity present in p13 beads or anti-cdc2 immunoprecipitates are seen in parallel with the accumulation of cells at G2, arguing that the arrest is not due to a failure to activate cdc2 as a histone H1 kinase. These results suggest that other changes in serine/threonine protein phosphorylation besides those involved in activation of cdc2 as a histone H1 kinase may be necessary for proper G2-M transition. Images PMID:1371461

Planas-Silva, M D; Means, A R

1992-01-01

115

Constitutive Smad signaling and Smad-dependent collagen gene expression in mouse embryonic fibroblasts lacking peroxisome proliferator-activated receptor-{gamma}  

SciTech Connect

Transforming growth factor-{beta} (TGF-{beta}), a potent inducer of collagen synthesis, is implicated in pathological fibrosis. Peroxisome proliferator-activated receptor-{gamma} (PPAR-{gamma}) is a nuclear hormone receptor that regulates adipogenesis and numerous other biological processes. Here, we demonstrate that collagen gene expression was markedly elevated in mouse embryonic fibroblasts (MEFs) lacking PPAR-{gamma} compared to heterozygous control MEFs. Treatment with the PPAR-{gamma} ligand 15d-PGJ{sub 2} failed to down-regulate collagen gene expression in PPAR-{gamma} null MEFs, whereas reconstitution of these cells with ectopic PPAR-{gamma} resulted in their normalization. Compared to control MEFs, PPAR-{gamma} null MEFs displayed elevated levels of the Type I TGF-{beta} receptor (T{beta}RI), and secreted more TGF-{beta}1 into the media. Furthermore, PPAR-{gamma} null MEFs showed constitutive phosphorylation of cellular Smad2 and Smad3, even in the absence of exogenous TGF-{beta}, which was abrogated by the ALK5 inhibitor SB431542. Constitutive Smad2/3 phosphorylation in PPAR-{gamma} null MEFs was associated with Smad3 binding to its cognate DNA recognition sequences, and interaction with coactivator p300 previously implicated in TGF-{beta} responses. Taken together, these results indicate that loss of PPAR-{gamma} in MEFs is associated with upregulation of collagen synthesis, and activation of intracellular Smad signal transduction, due, at least in part, to autocrine TGF-{beta} stimulation.

Ghosh, Asish K [Northwestern University Feinberg School of Medicine, Division of Rheumatology, 240 East Huron Street, Chicago, IL 60611 (United States)], E-mail: a-ghosh2@northwestern.edu; Wei, Jun; Wu, Minghua [Northwestern University Feinberg School of Medicine, Division of Rheumatology, 240 East Huron Street, Chicago, IL 60611 (United States); Varga, John [Northwestern University Feinberg School of Medicine, Division of Rheumatology, 240 East Huron Street, Chicago, IL 60611 (United States)], E-mail: j-varga@northwestern.edu

2008-09-19

116

The influence of caregiver singing and background music on vocally expressed emotions and moods in dementia care: A qualitative analysis  

Microsoft Academic Search

Background: Music and singing are considered to have a strong impact on human emotions. Such an effect has been demonstrated in caregiving contexts with dementia patients. Objectives: The aim of the study was to illuminate vocally expressed emotions and moods in the communication between caregivers and persons with severe dementia during morning care sessions. Design: Three types of caring sessions

Eva Gotell; Steven Brown; Sirkka-Liisa Ekman

117

Constitutive expression of Vitis vinifera thaumatin-like protein after in vitro selection and its role in anthracnose resistance  

Microsoft Academic Search

Anthracnose-resistant grapevine (Vitis vinifera L. cv. Chardonnay) plants were regenerated from embryogenic cultures that had been subjected to in vitro selection with culture filtrate of Elsinoe ampelina (de Bary) Shear. Three secreted proteins differentially expressed by in vitro-selected embryogenic cultures and regenerated plants were identified. An 8-kDa protein was identified as a lipid-transfer protein (LTP) by N-terminal amino acid sequence

Subramanian JayasankarA; Zhijian Li; Dennis J. Gray

2003-01-01

118

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

PubMed Central

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 as evaluated by in-situ PLA and IFA, and confirmed by Western blotting. The recombinant full-length and truncated proteins induced a high level of antibodies in mink, evaluated by ELISA, demonstrating their immunogenicity. In a challenge experiment in mink, a reduction in presentation clinical signs and virus 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. PMID:24376619

Bidokhti, Mehdi R. M.; Ullman, Karin; Jensen, Trine H.; Chriél, Mariann; Mottahedin, Amin; Munir, Muhammad; Andersson, Anna Maria; Detournay, Olivier; Hammer, Anne Sofie; Baule, Claudia

2013-01-01

119

Constitutive expression of the SAP1 gene from willow (Salix discolor) causes early flowering in Arabidopsis thaliana.  

PubMed

SAP1-1 and SAP1-2 were isolated from the male reproductive buds of willow (Salix discolor, clone S365). SAP1-1 differs from SAP1-2 based on a few nucleotide substitutions, but the sizes of their full-length cDNAs are identical. The deduced amino acid sequences of SAP1-1 and SAP1-2 were 98% similar and contain the same C-terminal amino acid motif "GYGA" like that of PTAP1-2 from Populus trichocarpa. The expression patterns of SAP1 in various parts of the male reproductive buds of S. discolor implicate this gene in the formation of the inflorescence meristems, bracts, and floral meristems. To characterize the functions of SAP1, we assessed Arabidopsis thaliana transformed with 35S: :SAP1-1. A total of 52 transgenic T1 lines were obtained, and a 3:1 segregation ratio was obtained in the T2 generation of each line. In the T3 generation, five homozygous transgenic lines were obtained, which were used for further analysis. Screening of transgenic lines was greatly facilitated by the detection of GFP expression starting with germinating seeds. Phenotypes of the homozygous transgenic lines included early flowering, conversion of inflorescence branches to solitary flowers, formation of terminal flowers, and formation of flowers with greater number of petals, stamens, and pistils. Northern analysis showed similar expression levels in all five lines. This study provides the first functional analysis of an APETALA1 (AP1)/SQUAMOSA (SQUA) homolog from a dioecious species and suggests that SAP1 is a homolog of the AP1/SQUA gene. PMID:16228224

Fernando, Danilo D; Zhang, Shiliang

2006-01-01

120

Constitutive and modulated expression of the human alpha 1 antitrypsin gene. Different transcriptional initiation sites used in three different cell types.  

PubMed Central

alpha 1-Antitrypsin (alpha 1 AT) is plasma glycoprotein that constitutes the principle inhibitor of neutrophil elastase in tissue fluids. It has been considered a prototype for liver-derived acute phase proteins in that its concentration in plasma increases three- to fourfold during the host response to inflammation/tissue injury. However, recent studies have shown that alpha 1 AT is expressed in several types of extrahepatic cells, including mononuclear phagocytes and enterocytes, and that there are distinct transcriptional units used in hepatocytes and at least one extra-hepatic cell type, blood monocytes. In this study, we have used a combination of ribonuclease protection assays, primer elongation analysis, and transcriptional run-on assays to further characterize mechanisms of basal and modulated alpha 1 AT gene expression in hepatocytes, enterocytes, and macrophages. The hepatoma cell line HepG2, intestinal epithelial cell line Caco2, and primary cultures of human peripheral blood monocytes were used as examples of the cell types. The results indicate that there are three macrophage-specific transcriptional initiation sites upstream from a single hepatocyte-specific transcriptional initiation site. Macrophages use these sites during basal and modulated expression. Hepatoma cells use the hepatocyte-specific transcriptional initiation site during basal and modulated expression but also switch on transcription from the upstream macrophage transcriptional initiation sites during modulation by the acute phase mediator interleukin 6 (IL-6). Caco2 cells use the hepatocyte-specific transcriptional initiation site during basal expression. There is a marked increase in the use of this site and an increase in the rate of transcriptional elongation of alpha 1 AT mRNA during differentiation of Caco2 cells from crypt-type to villous-type enterocytes. Caco2 cells also switch on transcription from the upstream macrophage transcriptional initiation sites during modulation by IL-6. These results provide further evidence that there are differences in the mechanisms of constitutive and regulated expression of the alpha 1 AT gene in at least three different cell types, HepG2-derived hepatocytes, Caco2-derived enterocytes and mononuclear phagocytes. Images PMID:1556183

Hafeez, W; Ciliberto, G; Perlmutter, D H

1992-01-01

121

Constitutive expression of the cytochrome P450 EthABCD monooxygenase system enables degradation of synthetic dialkyl ethers in Aquincola tertiaricarbonis L108.  

PubMed

In Rhodococcus ruber IFP 2001, Rhodococcus zopfii IFP 2005, and Gordonia sp. strain IFP 2009, the cytochrome P450 monooxygenase EthABCD catalyzes hydroxylation of methoxy and ethoxy residues in the fuel oxygenates methyl tert-butyl ether (MTBE), ethyl tert-butyl ether (ETBE), and tert-amyl methyl ether (TAME). The expression of the IS3-type transposase-flanked eth genes is ETBE dependent and controlled by the regulator EthR (C. Malandain et al., FEMS Microbiol. Ecol. 72:289-296, 2010). In contrast, we demonstrated by reverse transcription-quantitative PCR (RT-qPCR) that the betaproteobacterium Aquincola tertiaricarbonis L108, which possesses the ethABCD genes but lacks ethR, constitutively expresses the P450 system at high levels even when growing on nonether substrates, such as glucose. The mutant strain A. tertiaricarbonis L10, which is unable to degrade dialkyl ethers, resulted from a transposition event mediated by a rolling-circle IS91-type element flanking the eth gene cluster in the wild-type strain L108. The constitutive expression of Eth monooxygenase is likely initiated by the housekeeping sigma factor ?(70), as indicated by the presence in strain L108 of characteristic -10 and -35 binding sites upstream of ethA which are lacking in strain IFP 2001. This enables efficient degradation of diethyl ether, diisopropyl ether, MTBE, ETBE, TAME, and tert-amyl ethyl ether (TAEE) without any lag phase in strain L108. However, ethers with larger residues, n-hexyl methyl ether, tetrahydrofuran, and alkyl aryl ethers, were not attacked by the Eth system at significant rates in resting-cell experiments, indicating that the residue in the ether molecule which is not hydroxylated also contributes to the determination of substrate specificity. PMID:23354715

Schuster, Judith; Purswani, Jessica; Breuer, Uta; Pozo, Clementina; Harms, Hauke; Müller, Roland H; Rohwerder, Thore

2013-04-01

122

Constitutive Expression of the Cytochrome P450 EthABCD Monooxygenase System Enables Degradation of Synthetic Dialkyl Ethers in Aquincola tertiaricarbonis L108  

PubMed Central

In Rhodococcus ruber IFP 2001, Rhodococcus zopfii IFP 2005, and Gordonia sp. strain IFP 2009, the cytochrome P450 monooxygenase EthABCD catalyzes hydroxylation of methoxy and ethoxy residues in the fuel oxygenates methyl tert-butyl ether (MTBE), ethyl tert-butyl ether (ETBE), and tert-amyl methyl ether (TAME). The expression of the IS3-type transposase-flanked eth genes is ETBE dependent and controlled by the regulator EthR (C. Malandain et al., FEMS Microbiol. Ecol. 72:289–296, 2010). In contrast, we demonstrated by reverse transcription-quantitative PCR (RT-qPCR) that the betaproteobacterium Aquincola tertiaricarbonis L108, which possesses the ethABCD genes but lacks ethR, constitutively expresses the P450 system at high levels even when growing on nonether substrates, such as glucose. The mutant strain A. tertiaricarbonis L10, which is unable to degrade dialkyl ethers, resulted from a transposition event mediated by a rolling-circle IS91-type element flanking the eth gene cluster in the wild-type strain L108. The constitutive expression of Eth monooxygenase is likely initiated by the housekeeping sigma factor ?70, as indicated by the presence in strain L108 of characteristic ?10 and ?35 binding sites upstream of ethA which are lacking in strain IFP 2001. This enables efficient degradation of diethyl ether, diisopropyl ether, MTBE, ETBE, TAME, and tert-amyl ethyl ether (TAEE) without any lag phase in strain L108. However, ethers with larger residues, n-hexyl methyl ether, tetrahydrofuran, and alkyl aryl ethers, were not attacked by the Eth system at significant rates in resting-cell experiments, indicating that the residue in the ether molecule which is not hydroxylated also contributes to the determination of substrate specificity. PMID:23354715

Schuster, Judith; Purswani, Jessica; Breuer, Uta; Pozo, Clementina; Harms, Hauke; Müller, Roland H.

2013-01-01

123

Increased Na+/Ca2+ Exchanger Expression/Activity Constitutes a Point of Inflection in the Progression to Heart Failure of Hypertensive Rats  

PubMed Central

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. Conclusions: 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. PMID:24781001

Rodriguez, Jesica S.; Velez Rueda, J. Omar; Salas, Margarita; Becerra, Romina; Di Carlo, Mariano N.; Said, Matilde; Vittone, Leticia; Rinaldi, Gustavo; Portiansky, Enrique L.; Mundińa-Weilenmann, Cecilia; Palomeque, Julieta; Mattiazzi, Alicia

2014-01-01

124

DNA binding characteristics of RegA. A constitutively active anaerobic activator of photosynthesis gene expression in Rhodobacter capsulatus.  

PubMed

In the purple non-sulfur bacterium Rhodobacter capsulatus, RegA and RegB comprise a two-component regulatory system that is required for maximal anaerobic transcription of key photosynthesis genes. RegB is a sensor kinase that uses ATP to phosphorylate its cognate response regulator, RegA. The mechanism under which RegA approximately P influences transcription of target genes has been unclear given that past attempts to demonstrate DNA binding activity by isolated RegA have failed. This led to a model invoking a role for RegA approximately P as an intermediate in a more complex multicomponent phosphoryl transfer cascade. In the present study, we describe the isolation of a mutant version of RegA (RegA*) which promotes high level expression of photosynthesis genes independent of RegB. DNase I footprint analyses show that purified RegA* binds to the promoters of the puf and puc operons at locations that are consistent with RegA functioning as a transcriptional activator for these operons. We conclude that RegA functions, like most members of the response regulator family, as a DNA-binding protein that directly affects the expression of its target genes. PMID:9660820

Du, S; Bird, T H; Bauer, C E

1998-07-17

125

Molecular cloning and characterization of PtrLAR3, a gene encoding leucoanthocyanidin reductase from Populus trichocarpa, and its constitutive expression enhances fungal resistance in transgenic plants  

PubMed Central

The flavonoid-derived proanthocyanidins (PAs) are one class of the major defence phenolics in poplar leaves. Transcriptional activation of PA biosynthetic genes, resulting in PA accumulation in leaves, was detected following infection by the fungal Marssonina brunnea f.sp. multigermtubi using digital gene expression analysis. In order to study PA biosynthesis and its induction by fungi, a putative leucoanthocyanidin reductase gene, PtrLAR3, was isolated from Populus trichocarpa. Sequence comparison of PtrLAR3 with other known leucoanthocyanidin reductase proteins revealed high amino acid sequence similarity. Semi-quantitative reverse-transcription (RT) PCR and quantitative real-time PCR analysis demonstrated that PtrLAR3 was expressed in various tissues and the highest level of expression was observed in roots. Overexpression of PtrLAR3 in Chinese white poplar (Populus tomentosa Carr.) led to a significant plant-wide increase in PA levels. In vitro assays showed that crude leaf extracts from 35S:PtrLAR3 transformants were able to inhibit significantly the hyphal growth of M. brunnea f.sp. multigermtubi compared to the extracts from control plants. The transgenic 35S:PtrLAR3 poplar plants displayed a significant (P < 0.05) reduction in their disease symptoms compared with the control. RT-PCR analysis showed that PtrLAR3 expression was up-regulated in all transformants. These results suggested that constitutive expression of endogenous PtrLAR3 could be exploited to improve resistance to fungal pathogens in poplar. PMID:22268151

Yuan, Li; Wang, Lijun; Han, Zujing; Jiang, Yuanzhong; Zhao, Lili; Liu, Hong; Yang, Li; Luo, Keming

2012-01-01

126

The nuclear receptors pregnane X receptor and constitutive androstane receptor contribute to the impact of fipronil on hepatic gene expression linked to thyroid hormone metabolism.  

PubMed

Fipronil is described as a thyroid disruptor in rat. Based on the hypothesis that this results from a perturbation of hepatic thyroid hormone metabolism, our goal was to investigate the pathways involved in fipronil-induced liver gene expression regulations. First, we performed a microarray screening in the liver of rats treated with fipronil or vehicle. Fipronil treatment led to the upregulation of several genes involved in the metabolism of xenobiotics, including the cytochrome P450 Cyp2b1, Cyp2b2 and Cyp3a1, the carboxylesterases Ces2 and Ces6, the phase II enzymes Ugt1a1, Sult1b1 and Gsta2, and the membrane transporters Abcc2, Abcc3, Abcg5, Abcg8, Slco1a1 and Slco1a4. Based on a large overlap with the target genes of constitutive androstane receptor (CAR) and pregnane X receptor (PXR), we postulated that these two nuclear receptors are involved in mediating the effects of fipronil on liver gene expression in rodents. We controlled that liver gene expression changes induced by fipronil were generally reproduced in mice, and then studied the effects of fipronil in wild-type, CAR- and PXR-deficient mice. For most of the genes studied, the gene expression modulations were abolished in the liver of PXR-deficient mice and were reduced in the liver of CAR-deficient mice. However, CAR and PXR activation in mouse liver was not associated with a marked increase of thyroid hormone clearance, as observed in rat. Nevertheless, our data clearly indicate that PXR and CAR are key modulators of the hepatic gene expression profile following fipronil treatment which, in rats, may contribute to increase thyroid hormone clearance. PMID:23962444

Roques, Béatrice B; Leghait, Julien; Lacroix, Marlčne Z; Lasserre, Frédéric; Pineau, Thierry; Viguié, Catherine; Martin, Pascal G P

2013-10-01

127

Selection for low or high primary dormancy in Lolium rigidum Gaud seeds results in constitutive differences in stress protein expression and peroxidase activity  

PubMed Central

Seed dormancy in wild Lolium rigidum Gaud (annual ryegrass) populations is highly variable and not well characterized at the biochemical level. To identify some of the determinants of dormancy level in these seeds, the proteomes of subpopulations selected for low and high levels of primary dormancy were compared by two-dimensional polyacrylamide gel electrophoresis of extracts from mature, dry seeds. High-dormancy seeds showed higher expression of small heat shock proteins, enolase, and glyoxalase I than the low-dormancy seeds. The functional relevance of these differences in protein expression was confirmed by the fact that high-dormancy seeds were more tolerant to high temperatures imposed at imbibition and had consistently higher glyoxalase I activity over 0–42?d dark stratification. Higher expression of a putative glutathione peroxidase in low-dormancy seeds was not accompanied by higher activity, but these seeds had a slightly more oxidized glutathione pool and higher total peroxidase activity. Overall, these biochemical and physiological differences suggest that L. rigidum seeds selected for low dormancy are more prepared for rapid germination via peroxidase-mediated cell wall weakening, whilst seeds selected for high dormancy are constitutively prepared to survive environmental stresses, even in the absence of stress during seed development. PMID:20974739

Goggin, Danica E.; Powles, Stephen B.; Steadman, Kathryn J.

2011-01-01

128

Biotic and abiotic stress tolerance in transgenic tomatoes by constitutive expression of S-adenosylmethionine decarboxylase gene.  

PubMed

Recent findings have implicated the role of polyamines (putrescine, spermidine and spermine) in stress tolerance. Therefore, the present work was carried out with the goal of generating transgenic tomato plants with human S-adenosylmethionine decarboxylase (samdc) gene, a key gene involved in biosynthesis of polyamines, viz. spermidine and spermine and evaluating the transgenic plants for tolerance to both biotic and abiotic stresses. Several putative transgenic tomato plants with normal phenotype were obtained, and the transgene integration and expression was validated by PCR, Southern blot analysis and RT-PCR analysis, respectively. The transgenic plants exhibited high levels of polyamines as compared to the untransformed control plants. They also showed increased resistance against two important fungal pathogens of tomato, the wilt causing Fusarium oxysporum and the early blight causing Alternaria solani and tolerance to multiple abiotic stresses such as salinity, drought, cold and high temperature. These results suggest that engineering polyamine accumulation can confer tolerance to both biotic and abiotic stresses in plants. PMID:23573001

Hazarika, Pranjal; Rajam, Manchikatla Venkat

2011-04-01

129

Constitutive expression of Gs?(R201C) in mice produces a heritable, direct replica of human fibrous dysplasia bone pathology and demonstrates its natural history.  

PubMed

Fibrous dysplasia of bone (FD) is a crippling skeletal disease associated with postzygotic mutations (R201C, R201H) of the gene encoding the ? subunit of the stimulatory G protein, Gs. By causing a characteristic structural subversion of bone and bone marrow, the disease results in deformity, hypomineralization, and fracture of the affected bones, with severe morbidity arising in childhood or adolescence. Lack of inheritance of the disease in humans is thought to reflect embryonic lethality of germline-transmitted activating Gs? mutations, which would only survive through somatic mosaicism. We have generated multiple lines of mice that express Gs?(R201C) constitutively and develop an inherited, histopathologically exact replica of human FD. Robust transgene expression in neonatal and embryonic tissues and embryonic stem (ES) cells were associated with normal development of skeletal tissues and differentiation of skeletal cells. As in humans, FD lesions in mice developed only in the postnatal life; a defined spatial and temporal pattern characterized the onset and progression of lesions across the skeleton. In individual bones, lesions developed through a sequence of three distinct histopathological stages: a primary modeling phase defined by endosteal/medullary excess bone formation and normal resorption; a secondary phase, with excess, inappropriate remodeling; and a tertiary fibrous dysplastic phase, which reproduced a full-blown replica of the human bone pathology in mice of age ?1 year. Gs? mutations are sufficient to cause FD, and are per se compatible with germline transmission and normal embryonic development in mice. Our novel murine lines constitute the first model of FD. PMID:24764158

Saggio, Isabella; Remoli, Cristina; Spica, Emanuela; Cersosimo, Stefania; Sacchetti, Benedetto; Robey, Pamela G; Holmbeck, Kenn; Cumano, Ana; Boyde, Alan; Bianco, Paolo; Riminucci, Mara

2014-11-01

130

The Constitution Community  

NSDL National Science Digital Library

A new addition to the National Archives and Records Administration (NARA) Digital Classroom site (see the November 18, 199 Scout Report for Social Sciences), the Constitution Community is a partnership between classroom teachers and education specialists from the NARA. Like other Digital Classroom projects, the Constitution Community offers sample lesson plans, correlated to the National History Standards, that make use of original documents and images from American history. In addition to the lesson plan, each also includes background information, a number of suggested teaching activities, and its connection to the US Constitution The lessons are grouped by eight historical eras, covering the American Revolution to the Great Depression to the Civil Rights Act. Highlights include The Growth of Regionalism, The Civil War as Photographed by Mathew Brady, FDR's First Inaugural Address: Declaring "War" on the Great Depression, and the Founding Documents of the Peace Corps.

131

Genetic background influences adaptation to cardiac hypertrophy and Ca2+ handling gene expression  

PubMed Central

Genetic variability has a profound effect on the development of cardiac hypertrophy in response to stress. Consequently, using a variety of inbred mouse strains with known genetic profiles may be powerful models for studying the response to cardiovascular stress. To explore this approach we looked at male C57BL/6J and 129/SvJ mice. Hemodynamic analyses of left ventricular pressures (LVPs) indicated significant differences in 129/SvJ and C57BL/6J mice that implied altered Ca2+ handling. Specifically, 129/SvJ mice demonstrated reduced rates of relaxation and insensitivity to dobutamine (Db). We hypothesized that altered expression of genes controlling the influx and efflux of Ca2+ from the sarcoplasmic reticulum (SR) was responsible and investigated the expression of several genes involved in maintaining the intracellular and sarcoluminal Ca2+ concentration using quantitative real-time PCR analyses (qRT-PCR). We observed significant differences in baseline gene expression as well as different responses in expression to isoproterenol (ISO) challenge. In untreated control animals, 129/SvJ mice expressed 1.68× more ryanodine receptor 2(Ryr2) mRNA than C57BL/6J mice but only 0.37× as much calsequestrin 2 (Casq2). After treatment with ISO, sarco(endo)plasmic reticulum Ca2+-ATPase(Serca2) expression was reduced nearly two-fold in 129/SvJ while expression in C57BL/6J was stable. Interestingly, ? (1) adrenergic receptor(Adrb1) expression was lower in 129/SvJ compared to C57BL/6J at baseline and lower in both strains after treatment. Metabolically, the brain isoform of creatine kinase (Ckb) was up-regulated in response to ISO in C57BL/6J but not in 129/SvJ. These data suggest that the two strains of mice regulate Ca2+ homeostasis via different mechanisms and may be useful in developing personalized therapies in human patients. PMID:23508205

Waters, Steve B.; Diak, Douglass M.; Zuckermann, Matthew; Goldspink, Paul H.; Leoni, Lara; Roman, Brian B.

2013-01-01

132

The effects of a constitutive expression of transforming growth factor-alpha on the growth of MCF-7 human breast cancer cells in vitro and in vivo.  

PubMed

It has been suggested that transforming growth factor-alpha (TGF-alpha) is a mitogenic autocrine growth factor for human breast cancer cells, responsible for mediating the mitogenic effects of 17 beta-estradiol (E2) in responsive cells. To test this hypothesis we have introduced eukaryotic expression vectors directing the expression of TGF-alpha mRNA into E2-responsive MCF-7 human breast cancer cells. Transfected cells produce levels of TGF-alpha equivalent to or greater than those produced by both E2-stimulated MCF-7 cells and hormone-independent MDA-MB-231 cells. One transfected clone (H8) secretes sufficient TGF-alpha to fully down-regulate EGF-R expression. However, both of the transfected clones that constitutively secrete elevated levels of TGF-alpha (A8 and H8) respond to E2 stimulation in vitro by increasing the rate of cellular proliferation and inducing PGR synthesis. The basal proliferative capacity of H8 and A8 cells is equivalent to that of the parental cells and to cells transfected only with the G418 (neomycin) resistance gene. Furthermore, the TGF-alpha cDNA-transfected clones do not form tumors in ovariectomized athymic nude mice without E2 supplementation. Thus, the precise role of TGF-alpha in mediating either the in vivo or the in vitro mitogenic effects of E2 in MCF-7 human breast cancer cells remains unclear. While TGF-alpha expression may be essential, it is not sufficient alone to induce the fully E2-independent phenotype. Thus, TGF-alpha may function in combination with other E2-induced growth factors to control breast cancer proliferation and tumorigenesis. PMID:2710138

Clarke, R; Brünner, N; Katz, D; Glanz, P; Dickson, R B; Lippman, M E; Kern, F G

1989-02-01

133

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

Microsoft Academic Search

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

Laura S. Robertson; Luke R. Iwanowicz; Jamie Marie Marranca

2009-01-01

134

Bone-marrow derived hematopoietic stem/progenitor cells express multiple isoforms of NADPH oxidase and produce constitutively reactive oxygen species.  

PubMed

Consolidated evidence highlights the importance of redox signalling in poising the balance between self-renewal and differentiation in adult stem cells. The present study shows that human hematopoietic stem/progenitor cells (HSCs) constitutively generate low levels of hydrogen peroxide whose production is inhibited by DPI, apocynin, catalase, and LY294002 and scarcely stimulated by PMA. Moreover, it is shown that HSCs express at the mRNA and protein levels the catalytic subunits of NOX1, NOX2, and NOX4 isoforms of the NADPH oxidase family along with the complete battery of the regulatory subunits p22, p40, p47, p67, rac1, rac2, NOXO1, and NOXA1 as well as the splicing variant NOX2s and that the three NOX isoforms are largely co-expressed in the same HSC. These findings are interpreted in terms of a positive feed-back mechanism of NOXs activation enabling a fine tuning of the ROS level to be possibly used in redox-mediated signalling for growth and differentiation of HSCs. PMID:17204244

Piccoli, Claudia; D'Aprile, Annamaria; Ripoli, Maria; Scrima, Rosella; Lecce, Lucia; Boffoli, Domenico; Tabilio, Antonio; Capitanio, Nazzareno

2007-02-23

135

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.  

PubMed

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. PMID:25044501

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

2014-10-01

136

Efficient constitutive expression of chitinase in the mother cell of Bacillus thuringiensis and its potential to enhance the toxicity of Cry1Ac protoxin.  

PubMed

Previous studies revealed that chitinase could enhance the insecticidal activity of Bacillus thuringiensis and it has been used in combination with B. thuringiensis widely. However, the expression of B. thuringiensis chitinase is rather low and needs induction by chitin, which limits its field application. It would make sense to constitutively express the chitinase at a sufficiently high level to offer advantages in biological control of pests. In this study, a signal peptide-encoding sequence-deleted chitinase gene from B. thuringiensis strain 4.0718 under the control of dual overlapping promoters plus Shine-Dalgarno sequence and terminator sequence of cry1Ac3 gene was cloned into shuttle vector pHT315 and introduced into an acrystalliferous B. thuringiensis strain Cry(-)B. The recombinant plasmid was stably maintained over 240 generations in Cry(-)B. Chitinase was overexpressed within the sporangial mother cells in the form of spherical crystal-like inclusion bodies. The chitinase inclusions could be solubilized and exhibit chitinolytic activity in 30 mmol l(-1) Na(2)CO(3)-0.2% beta-mercaptoethanol buffer at a wide range of alkaline pH values, and what's more, the chitinase inclusions potentiated the insecticidal effect of Cry1Ac protoxin when used against larvae of Spodoptera exigua and Helicoverpa armigera. PMID:19277644

Hu, S B; Liu, P; Ding, X Z; Yan, L; Sun, Y J; Zhang, Y M; Li, W P; Xia, L Q

2009-04-01

137

Quantitative expression profiling of G-protein-coupled receptors (GPCRs) in metastatic melanoma: the constitutively active orphan GPCR GPR18 as novel drug target.  

PubMed

G-protein-coupled receptors (GPCRs) have been implicated in the tumorigenesis and metastasis of human cancers and are considered amongst the most desirable targets for drug development. Utilizing a robust quantitative PCR array, we quantified expression of 94 human GPCRs, including 75 orphan GPCRs and 19 chemokine receptors, and 36 chemokine ligands, in 40 melanoma metastases from different individuals and benign nevi. Inter-metastatic site comparison revealed that orphan GPR174 and CCL28 are statistically significantly overexpressed in subcutaneous metastases, while P2RY5 is overexpressed in brain metastases. Comparison between metastases (all three metastatic sites) and benign nevi revealed that 16 genes, including six orphan receptors (GPR18, GPR34, GPR119, GPR160, GPR183 and P2RY10) and chemokine receptors CCR5, CXCR4, and CXCR6, were statistically significantly differentially expressed. Subsequent functional experiments in yeast and melanoma cells indicate that GPR18, the most abundantly overexpressed orphan GPCR in all melanoma metastases, is constitutively active and inhibits apoptosis, indicating an important role for GPR18 in tumor cell survival. GPR18 and five other orphan GPCRs with yet unknown biological function may be considered potential novel anticancer targets in metastatic melanoma. PMID:20880198

Qin, Y; Verdegaal, E M E; Siderius, M; Bebelman, J P; Smit, M J; Leurs, R; Willemze, R; Tensen, C P; Osanto, S

2011-02-01

138

Interleukin-22 binding protein (IL-22BP) is constitutively expressed by a subset of conventional dendritic cells and is strongly induced by retinoic acid  

PubMed Central

IL-22 is mainly produced at barrier surfaces by T cells and innate lymphoid cells and is crucial to maintain epithelial integrity. However, dysregulated IL-22 action leads to deleterious inflammation and is involved in diseases such as psoriasis, intestinal inflammation and cancer. IL-22BP is a soluble inhibitory IL-22 receptor and may represent a crucial regulator of IL-22. We show both in rats and mice that, in the steady state, the main source of IL-22BP is constituted by a subset of conventional dendritic cells (DC) in lymphoid and non lymphoid tissues. In mouse intestine, IL-22BP was specifically expressed in lamina propria CD103+CD11b+ DC. In humans, IL-22BP was expressed in immature monocyte-derived DC (MDDC) and strongly induced by retinoic acid (RA) but dramatically reduced upon maturation. Our data suggest that a subset of immature DC may actively participate in the regulation of IL-22 activity in the gut by producing high levels of IL-22BP. PMID:23653115

Martin, JCJ; Bériou, G; Heslan, M; Chauvin, C; Utriainen, L; Aumeunier, A; Scott, CL; Mowat, A; Cerovic, V; Houston, SA; Leboeuf, M; Hubert, FX; Hémont, C; Merad, M; Milling, S; Josien, R

2014-01-01

139

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

PubMed

Extracytoplasmic function (ECF) sigma factors (?(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 ?(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. PMID:23059974

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

2013-02-01

140

Transient cardiac expression of constitutively active G?q leads to hypertrophy and dilated cardiomyopathy by calcineurin-dependent and independent pathways  

PubMed Central

Cardiac hypertrophy and dilatation can result from stimulation of signal transduction pathways mediated by heterotrimeric G proteins, especially Gq, whose ? subunit activates phospholipase C? (PLC?). We now report that transient, modest expression of a hemagglutinin (HA) epitope-tagged, constitutively active mutant of the Gq ? subunit (HA?*q) in hearts of transgenic mice is sufficient to induce cardiac hypertrophy and dilatation that continue to progress after the initiating stimulus becomes undetectable. At 2 weeks, HA?*q protein is expressed at less than 50% of endogenous ?q/11, and the transgenic hearts are essentially normal morphologically. Although HA?*q protein declines at 4 weeks and is undetectable by 10 weeks, the animals develop cardiac hypertrophy and dilatation and die between 8 and 30 weeks in heart failure. As the pathology develops, endogenous ?q/11 rises (2.9-fold in atria; 1.8-fold in ventricles). At 2 weeks, basal PLC activity is increased 9- to 10-fold in atria but not ventricles. By 10 weeks, it is elevated in both, presumably because of the rise in endogenous ?q/11. We conclude that the pathological changes initiated by early, transient HA?*q expression are maintained in part by compensatory changes in signal transduction and other pathways. Cyclosporin A (CsA) prevents hypertrophy caused by activation of calcineurin [Molkentin, J. D., Lu, J.-R., Antos, C. L., Markham, B., Richardson, J., Robbins, J., Grant, S. R. & Olson, E. N. (1998) Cell 93, 215–228]. Because HA?*q acts upstream of calcineurin, we hypothesized that HA?*q might initiate additional pathways leading to hypertrophy and dilatation. Treating HA?*q mice with CsA diminished some, but not all, aspects of the hypertrophic phenotype, suggesting that multiple pathways are involved. PMID:9811897

Mende, U.; Kagen, A.; Cohen, A.; Aramburu, J.; Schoen, F. J.; Neer, E. J.

1998-01-01

141

Constitutive expression of selected genes from the pentose phosphate and aromatic pathways increases the shikimic acid yield in high-glucose batch cultures of an Escherichia coli strain lacking PTS and pykF  

PubMed Central

Background During the last two decades many efforts have been directed towards obtaining efficient microbial processes for the production of shikimic acid (SA); however, feeding high amounts of substrate to increase the titer of this compound has invariably rendered low conversion yields, leaving room for improvement of the producing strains. In this work we report an alternative platform to overproduce SA in a laboratory-evolved Escherichia coli strain, based on plasmid-driven constitutive expression of six genes selected from the pentose phosphate and aromatic amino acid pathways, artificially arranged as an operon. Production strains also carried inactivated genes coding for phosphotransferase system components (ptsHIcrr), shikimate kinases I and II (aroK and aroL), pyruvate kinase I (pykF) and the lactose operon repressor (lacI). Results The strong and constitutive expression of the constructed operon permitted SA production from the beginning of the cultures, as evidenced in 1 L batch-mode fermentors starting with high concentrations of glucose and yeast extract. Inactivation of the pykF gene improved SA production under the evaluated conditions by increasing the titer, yield and productivity of this metabolite compared to the isogenic pykF+ strain. The best producing strain accumulated up to 43 g/L of SA in 30 h and relatively low concentrations of acetate and aromatic byproducts were detected, with SA accounting for 80% of the produced aromatic compounds. These results were consistent with high expression levels of the glycolytic pathway and synthetic operon genes from the beginning of fermentations, as revealed by transcriptomic analysis. Despite the consumption of 100 g/L of glucose, the yields on glucose of SA and of total aromatic compounds were about 50% and 60% of the theoretical maximum, respectively. The obtained yields and specific production and consumption rates proved to be constant with three different substrate concentrations. Conclusions The developed production system allowed continuous SA accumulation until glucose exhaustion and eliminated the requirement for culture inducers. The obtained SA titers and yields represent the highest reported values for a high-substrate batch process, postulating the strategy described in this report as an interesting alternative to the traditionally employed fed-batch processes for SA production. PMID:24079972

2013-01-01

142

What Is a Constitution?  

ERIC Educational Resources Information Center

Provides a lesson plan designed to help students better understand the concept of a constitution, distinguish constitutional law from statutory law, and recognize examples of constitutional government. (BSR)

OAH Magazine of History, 1988

1988-01-01

143

Transcript Expression Patterns Illuminate the Mechanistic Background of Hormesis in Caenorhabditis Elegans Maupas  

PubMed Central

The animal model Caenorhabditis elegans was employed to study polyphenol- and humic substances-induced hormetic changes in lifespan. A detailed insight into the underlying mechanism of hormesis was uncovered by applying whole genome DNA microarray experimentation over a range of quercetin (Q), tannic acid (TA), and humic substances (HuminFeed®, HF) concentrations. The transcriptional response to all exposures followed a non-linear mode which highlighted differential signaling and metabolic pathways. While low Q concentrations regulated processes improving the health of the nematodes, higher concentrations extended lifespan and modulated substantially the global transcriptional response. Over-represented transcripts were notably part of the biotransformation process: enhanced catabolism of toxic intermediates possibly contributes to the lifespan extension. The regulation of transcription, Dauer entry, and nucleosome suggests the presence of distinct exposure dependent differences in transcription and signaling pathways. TA- and HF-mediated transcript expression patterns were overall similar to each other, but changed across the concentration range indicating that their transcriptional dynamics are complex and cannot be attributed to a simple adaptive response. In contrast, Q-mediated hormesis was well aligned to fit the definition of an adaptive response. Simple molecules are more likely to induce an adaptive response than more complex molecules.

Steinberg, Christian E.W.; Pietsch, Kerstin; Saul, Nadine; Menzel, Stefanie; Swain, Suresh C.; Stürzenbaum, Stephen R.; Menzel, Ralph

2013-01-01

144

Behavior of a cloned murine interferon alpha/beta receptor expressed in homospecific or heterospecific background.  

PubMed Central

A murine interferon (IFN) alpha/beta receptor was cloned from the IFN-sensitive L1210 cell line on the basis of its homology with the human receptor. A combination of methods that includes the screening of random-primed and oligo(dT)-primed cDNA libraries and polymerase chain reactions with a single-side specificity was used. At the amino acid level, the murine IFN-alpha/beta shows 46% identity with its human counterpart. Both human WISH cells presenting a low sensitivity to mouse IFN and a murine L1210 mutant subline that does not express the receptor have been stably transfected with the murine IFN-alpha/beta receptor. Whereas transfected human cells became sensitive to a limited number of mouse IFN-alpha/beta subtypes, the transfected murine L1210 mutant was found to be fully complemented and became sensitive to all mouse IFN-alpha/beta subtypes tested, including those that were not active on transfected human cells. These results strongly suggest that the receptor described here is implicated in the mediation of the activities of all murine IFN-alpha/beta subtypes. Images PMID:1533935

Uzé, G; Lutfalla, G; Bandu, M T; Proudhon, D; Mogensen, K E

1992-01-01

145

Characterization and constitutive expression of an acidic mesophilic endo-1,4-?-D-xylanohydrolase with high thermotolerance and catalytic efficiency in Pichia pastoris.  

PubMed

A putative endo-1,4-?-D-xylanohydrolase gene xyl11 from Aspergillus niger, encoding a 188-residue xylanase of glycosyl hydrolase family 11, was constitutively expressed in Pichia pastoris. The recombinant Xyl11 exhibited optimal activity at pH 5.0 and 50 °C, and displayed more than 68 % of the maximum activity over the temperature range 35-65 °C and 33 % over the pH range 2.2-7.0. It maintained more than 40 % of the original activity after incubation at 90 °C (pH 5.0) for 10 min and more than 75 % of the original activity after incubation at pH 2.2-11.0 (room temperature) for 2 h. The specific activity, K m and V max of purified Xyl11 were 22,253 U mg(-1), 6.57 mg ml(-1) and 51,546.4 ?mol min(-1) mg(-1). It could degrade xylan to a series of xylooligosaccharides and no xylose was detected. The recombinant enzyme with high stability and catalytic efficiency could work over wide ranges of pH and temperature and thus has the potential for various industrial applications. PMID:23670313

Guo, Ning; Zheng, Jia; Tian, Jian; Wu, Lishuang; Zhou, Hongbo

2013-11-01

146

Mature mRNAs accumulated in the nucleus are neither the molecules in transit to the cytoplasm nor constitute a stockpile for gene expression.  

PubMed Central

In higher eukaryotes, the regulation of pre-mRNA processing is still poorly known. The accumulation of various mature mRNAs, which can be observed in the nuclei of mammalian cells, is suggestive of a regulatory role of transport. However, the significance of these nuclear mRNA is presently unknown. We have used a tetracycline-regulated promoter to investigate the dynamics of these pools of mRNAs upon arrest of transcription. We observed, for beta-globin and LT-alpha genes, a slow disappearance of these mRNA from the nucleus, with an apparent half-life that is similar to their cytoplasmic half-life. In view of these dynamics, these mRNA cannot simply be mature mRNAs in transit to the cytoplasm. They could be mRNAs retained in the nucleus, provided that the regulation of mRNA stability is comparable in the nucleus and the cytoplasm. But, because of their limited stability, these nuclear mRNAs cannot constitute a significant stock for gene expression. Alternatively, they could reflect a bidirectional transport of mRNA, that is, to and from the cytoplasm, which would provide a direct explanation for the similarity in both compartments of their half-life and poly(A) tail shortening over time. PMID:10917593

Weil, D; Boutain, S; Audibert, A; Dautry, F

2000-01-01

147

Constitutive expression of a salinity-induced wheat WRKY transcription factor enhances salinity and ionic stress tolerance in transgenic Arabidopsis thaliana  

SciTech Connect

Highlights: •A class II WRKY transcription factor, TaWRKY79 was isolated and characterized. •TaWRKY79 was induced by NaCl or abscisic acid. •843 bp regulatory segment was sufficient to respond to ABA or NaCl treatment. •TaWRKY79 enhanced salinity and ionic tolerance while reduced sensitivity to ABA. •TaWRKY79 increased salinity and ionic tolerance in an ABA-dependent pathway. -- Abstract: The isolation and characterization of TaWRKY79, a wheat class II WRKY transcription factor, is described. Its 1297 bp coding region includes a 987 bp long open reading frame. TaWRKY79 was induced by stressing seedlings with either NaCl or abscisic acid (ABA). When a fusion between an 843 bp segment upstream of the TaWRKY79 coding sequence and GUS was introduced into Arabidopsis thaliana, GUS staining indicated that this upstream segment captured the sequence(s) required to respond to ABA or NaCl treatment. When TaWRKY79 was constitutively expressed as a transgene in A. thaliana, the transgenic plants showed an improved capacity to extend their primary root in the presence of either 100 mM NaCl, 10 mM LiCl or 2 ?M ABA. The inference was that TaWRKY79 enhanced the level of tolerance to both salinity and ionic stress, while reducing the level of sensitivity to ABA. The ABA-related genes ABA1, ABA2 ABI1 and ABI5 were all up-regulated in the TaWRKY79 transgenic plants, suggesting that the transcription factor operates in an ABA-dependent pathway.

Qin, Yuxiang, E-mail: yuxiangqin@126.com [Department of Biotechnology, University of Jinan, Jinan 250022 (China)] [Department of Biotechnology, University of Jinan, Jinan 250022 (China); Tian, Yanchen [The Key Laboratory of Plant Cell Engineering and Germplasm Innovation, Ministry of Education, School of Life Science, Shandong University, Jinan 250100 (China)] [The Key Laboratory of Plant Cell Engineering and Germplasm Innovation, Ministry of Education, School of Life Science, Shandong University, Jinan 250100 (China); Han, Lu; Yang, Xinchao [Department of Biotechnology, University of Jinan, Jinan 250022 (China)] [Department of Biotechnology, University of Jinan, Jinan 250022 (China)

2013-11-15

148

Morphoproteomics identifies constitutive activation of the mTORC2/Akt and NF-?B pathways and expressions of IGF-1R, Sirt1, COX-2, and FASN in peripheral T-cell lymphomas: pathogenetic implications and therapeutic options  

PubMed Central

Background: Gaining a better understanding of the molecular circuitries and pathways implicated in the malignant growth and biological behavior of T cell lymphomas may identify potential cellular targets with clinical therapeutic potential. The immunohistochemical characterization of key cellular proteins participating in these pathways can provide surrogate markers of biological activity. The mammalian target of rapamycin complex (mTORC) signaling pathway has been implicated in T-cell lymphopoiesis. The mTORC2 pathway involves downstream activation of nuclear factor (NF)-?B and p-Akt (Ser 473). Fatty acid synthase (FASN) and insulin-like growth factor-1 receptor (IGF-1R) are expressed upstream of the mTORC and NF-?B signaling pathways. Cyclooxygenase (COX)-2 products influence these pathways. Our goal was to use morphoproteomics to characterize the expression patterns of the proteins in various peripheral T-cell lymphomas. Design: Ten cases of peripheral T-cell lymphoma (PTCL) were examined for expression of proteins along the mTORC, Akt and NF-?B pathways and affiliated tumorigenic molecules. These included two angioimmunoblastic PTCL, one natural killer/PTCL, one anaplastic large PTCL, and six PTCL not otherwise specified. Immunostaining for phosphorylated (p) mTOR (Ser 2448), p-Akt (Ser 473), p-NF-?Bp65 (Ser 536), IGF-1R (Tyr1165/1166), silent mating type information regulation 2 homolog 1 (Sirt1), COX-2 and FASN was performed on paraffin-embedded tissue for each case. Percent expression was scored using bright-field microscopy with high expression designated as more than 50% of the cells with positive stain in the appropriate subcellular compartment. Results: All ten cases demonstrated nuclear staining for p-mTOR (Ser 2448) corresponding to mTORC 2, and all cases showed strong, diffuse nuclear staining for p-NF-?Bp65 (Ser 536). All ten also showed nuclear and cytoplasmic staining for p-Akt (Ser 473) and cytoplasmic staining for IGF-1R. High expressions for nuclear Sirt1, and cytoplasmic COX-2 and FASN were detected in 7, 9, and 8 out of 10 cases, respectively. Six out of 10 cases demonstrated high expression of all the mentioned markers. Conclusion: The constitutive activation of mTORC2, NF-?B, p-Akt and the concomitant expression of IGF-1R suggests convergence of these molecular pathways in T-cell lymphoma. The results of this study also suggest that mTORC2 may be a common denominator among this heterogeneous group of lymphomas. Interference of key nodes of this pathway may carry a clinical therapeutic benefit. Agents that may be considered based on existing data include: bortezomib to inhibit NF-?B pathway activation; metformin to inhibit both NF-?B and mTORC2 and histone deacteylase inhibitors to inhibit mTORC2 pathway signaling. Furthermore, panobinostat inhibits Sirt1 pathway when present, and celecoxib inhibits NF-?B pathway activation independent of COX2.

Quesada, Andrés E; Nguyen, Nghia D; Rios, Adan; Brown, Robert E

2014-01-01

149

Amending the Constitution: Current Proposals.  

ERIC Educational Resources Information Center

Provides background information on four constitutional amendment proposals: (1) flag protection amendment; (2) school prayer amendment; (3) balanced-budget amendment; and (4) victim rights amendment. Evaluates each in terms of timeliness, the support base, and the strength of their opposition. Includes questions for discussion. (CMK)

Schechter, Stephen L.

2000-01-01

150

Different strategies for reducing intestinal background radioactivity associated with imaging HSV1-tk expression using established radionucleoside probes  

PubMed Central

One limitation of HSV1-tk reporter PET imaging with nucleoside analogues is the high background radioactivity in the intestine. We hypothesized that endogenous expression of thymidine kinase in bacterial flora could phosphorylate and trap such radiotracers, contributing to the high radioactivity levels in the bowel and therefore explored different strategies to increase fecal elimination of radiotracer. Methods Intestinal radioactivity was assessed by in vivo microPET imaging and ex vivo tissue sampling following intravenous injection of 18F-FEAU, 124I-FIAU or 18F-FHBG in a germ-free mouse strain. We also explored the use of an osmotic laxative agent and/or a 100% enzymatically hydrolyzed liquid diet. Results No significant differences in intestinal radioactivity were observed between germ-free and normal mice. 18F-FHBG-derived intestinal radioactivity levels were higher than those of 18F-FEAU and 124I-FIAU; the intestine-to-blood ratio was more than 20-fold higher for 18F-FHBG than for 18F-FEAU and 124I-FIAU. The combination of Peptamen and Nulytely lowered intestinal radioactivity levels and increased (2.2-fold) the HSV1-tk transduced xenograft-to-intestine ratio for 18F-FEAU. Conclusions Intestinal bacteria in germ-free mice do not contribute to the high intestinal levels of radioactivity following injection of radionucleoside analogs. The combination of Peptamen and Nulytely increased radiotracer elimination by increasing bowel motility without inducing dehydration. PMID:20128998

Ruggiero, Alessandro; Brader, Peter; Serganova, Inna; Zanzonico, Pat; Cai, Shangde; Lipman, Neil S.; Hricak, Hedvig; Blasberg, Ronald G.

2011-01-01

151

The Constitutional Amendment Process  

ERIC Educational Resources Information Center

This article discusses the constitutional amendment process. Although the process is not described in great detail, Article V of the United States Constitution allows for and provides instruction on amending the Constitution. While the amendment process currently consists of six steps, the Constitution is nevertheless quite difficult to change.…

Chism, Kahlil

2005-01-01

152

SURVIVAL AND DEGRADATIVE CAPACITY OF PSEUDOMONAS PUTIDA INDUCED OR CONSTITUTIVELY EXPRESSING PLASMA-MEDIATED DEGRADATION OF 2,4-DICHLOROPHENOXYACETATE (TFD) IN SOIL  

EPA Science Inventory

The survival of genetically altered Pseudomonas putida strains harboring an inducible plasmid, PRO101, or a constitutive plasmid, PRO103, was compared. hese plasmids encode for the degradation of 2,4-dichlorophenoxyacetate (TFD) to 2-chloromaleylacetate, and the maintenance of ei...

153

SURVIVAL AND DEGRADATIVE CAPACITY OF PSEUDOMONAS PUTIDA INDUCED OR CONSTITUTIVELY EXPRESSING PLASMID-MEDIATED DEGRADATION OF 2,4-DICHLOROPHENOXYACETATE (TFD) IN SOIL  

EPA Science Inventory

Survival of genetically altered Pseudonomas putida strains harboring an inducible plasmid, pRO101, or a constitutive plasmid, pRO103, was compared. hese plasmids encoded for the degradation of 2,4-dichlorophenoxyacetate (TFD) to 2-chloromaleylacetate, and the maintenance of eithe...

154

Constitutive Activation of Nuclear Factor-E2-Related Factor 2 Induces Biotransformation Enzyme and Transporter Expression in Livers of Mice With Hepatocyte-Specific Deletion of Kelch-like ECH-associated protein 1  

PubMed Central

Chemicals that activate nuclear factor-E2-related factor-2 (Nrf2) often increase multidrug resistance-associated protein expression in liver. Hepatocyte-specific deletion of Kelch-like ECH-associated protein 1 (Keap1) activates Nrf2. Use of hepatocyte-specific Keap1 deletion represents a non-pharmacological method to determine whether constitutive Nrf2 activation upregulates liver transporter expression in vivo. The mRNA, protein expression and localization of several biotransformation and transporters was determined in livers of wild-type and hepatocyte-specific Keap1-null mice. Sulfotransferase 2a1/2, NADP(H):quinone oxidoreductase 1, Cytochrome P450 2b10, 3a11, and glutamate-cysteine ligase catalytic subunit expression was increased in livers of Keap1-null mice. Oatp1a1 expression was nearly abolished, as compared to that detected in livers of wild-type mice. By contrast, Mrp 1-5 mRNA and protein levels were increased in Keap1-null mouse livers, with Mrp4 expression being more than 15-fold higher than wild-types. In summary, Nrf2 has a significant role in affecting expression of Oatp and Mrp expression. PMID:21538727

Cheng, Qiuqiong; Taguchi, Keiko; Aleksunes, Lauren M.; Manautou, José E.; Cherrington, Nathan J.; Yamamoto, Masayuki; Slitt, Angela L.

2013-01-01

155

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.  

PubMed

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. PMID:23934571

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

2013-09-01

156

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.  

PubMed

Abstract 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 accumulating, 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 72% 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 6 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. PMID:23729566

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

2013-05-17

157

Escherichia coli cells expressing a mutant glyV (glycine tRNA) gene have a UVM-constitutive phenotype: implications for mechanisms underlying the mutA or mutC mutator effect.  

PubMed

Transfection of M13 single-stranded viral DNA bearing a 3,N4-ethenocytosine lesion into Escherichia coli cells pretreated with UV results in a significant elevation of mutagenesis at the lesion site compared to that observed in untreated cells. This response, termed UVM, for UV modulation of mutagenesis, is induced by a variety of DNA-damaging agents and is distinct from known cellular responses to DNA damage, including the SOS response. This report describes our observation, as a part of our investigation of the UVM phenomenon, that E. coli cells bearing a mutA or mutC allele display a UVM-constitutive phenotype. These mutator alleles were recently mapped (M. M. Slupska, C. Baikalov, R. Lloyd, and J. H. Miller, Proc. Natl. Acad. Sci. USA 93:4380-4385, 1996) to the glyV (mutA) and glyW (mutC) tRNA genes. Each mutant allele was shown to arise by an identical mutation in the anticodon sequence such that the mutant tRNAs could, in principle, mistranslate aspartate codons in mRNA as glycine at a low level. Because a UVM-constitutive phenotype resulting from a mutation in a tRNA gene was unexpected, we undertook a series of experiments designed to test whether the phenotype was indeed mediated by the expression of mutant glycine tRNAs. We placed either a wild-type or a mutant glyV gene under the control of a heterologous inducible promoter on a plasmid vector. E. coli cells expressing the mutant glyV gene displayed all three of the following phenotypes: (i) missense suppression of a test allele, (ii) a mutator phenotype measured by mutation to rifampin resistance, and (iii) a UVM-constitutive phenotype. These phenotypes were not associated with cells expressing the wild-type glyV gene or with cells in which the mutant allele was present but was not transcriptionally induced. These observations provide strong support for the idea that expression of mutant tRNA can confer a mutator phenotype, including the UVM-constitutive phenotype observed in mutA and mutC cells. However, our data imply that low-level mistranslation of the epsilon subunit of polymerase III probably does not account for the observed UVM-constitutive phenotype. Our results also indicate that mutA deltarecA double mutants display a normal UVM phenotype, suggesting that the mutA effect is recA dependent. The observations reported here raise a number of intriguing questions and raise the possibility that the UVM response is mediated through transient alteration of the replication environment. PMID:9393717

Murphy, H S; Humayun, M Z

1997-12-01

158

Escherichia coli cells expressing a mutant glyV (glycine tRNA) gene have a UVM-constitutive phenotype: implications for mechanisms underlying the mutA or mutC mutator effect.  

PubMed Central

Transfection of M13 single-stranded viral DNA bearing a 3,N4-ethenocytosine lesion into Escherichia coli cells pretreated with UV results in a significant elevation of mutagenesis at the lesion site compared to that observed in untreated cells. This response, termed UVM, for UV modulation of mutagenesis, is induced by a variety of DNA-damaging agents and is distinct from known cellular responses to DNA damage, including the SOS response. This report describes our observation, as a part of our investigation of the UVM phenomenon, that E. coli cells bearing a mutA or mutC allele display a UVM-constitutive phenotype. These mutator alleles were recently mapped (M. M. Slupska, C. Baikalov, R. Lloyd, and J. H. Miller, Proc. Natl. Acad. Sci. USA 93:4380-4385, 1996) to the glyV (mutA) and glyW (mutC) tRNA genes. Each mutant allele was shown to arise by an identical mutation in the anticodon sequence such that the mutant tRNAs could, in principle, mistranslate aspartate codons in mRNA as glycine at a low level. Because a UVM-constitutive phenotype resulting from a mutation in a tRNA gene was unexpected, we undertook a series of experiments designed to test whether the phenotype was indeed mediated by the expression of mutant glycine tRNAs. We placed either a wild-type or a mutant glyV gene under the control of a heterologous inducible promoter on a plasmid vector. E. coli cells expressing the mutant glyV gene displayed all three of the following phenotypes: (i) missense suppression of a test allele, (ii) a mutator phenotype measured by mutation to rifampin resistance, and (iii) a UVM-constitutive phenotype. These phenotypes were not associated with cells expressing the wild-type glyV gene or with cells in which the mutant allele was present but was not transcriptionally induced. These observations provide strong support for the idea that expression of mutant tRNA can confer a mutator phenotype, including the UVM-constitutive phenotype observed in mutA and mutC cells. However, our data imply that low-level mistranslation of the epsilon subunit of polymerase III probably does not account for the observed UVM-constitutive phenotype. Our results also indicate that mutA deltarecA double mutants display a normal UVM phenotype, suggesting that the mutA effect is recA dependent. The observations reported here raise a number of intriguing questions and raise the possibility that the UVM response is mediated through transient alteration of the replication environment. PMID:9393717

Murphy, H S; Humayun, M Z

1997-01-01

159

ALTERED SENSITIVITY OF THE MOUSE FETUS TO IMPAIRED PROSTATIC BUD FORMATION BY DIOXIN: INFLUENCE OF GENETIC BACKGROUND AND NULL EXPRESSION OF TGF-ALFA AND EGF  

EPA Science Inventory

Altered sensitivity of the mouse fetus to impaired prostatic bud formation by dioxin: Influence of genetic background and null expression of TGF and EGF. Rasmussen, N.T., Lin T-M., Fenton, S.E., Abbott, B.D. and R.E. Peterson. 2,3,7,8-Tetrachlorodibenzo-p-dioxin (TCDD)...

160

Regulation of Cell Growth and Cyclin D1 Expression by the Constitutively Active FRAP-p70s6K Pathway in Human Pancreatic Cancer Cells1  

Microsoft Academic Search

The FRAP-p70s6K signaling pathway was found to be constitutively phosphorylated\\/active in MiaPaCa-2 and Panc-1 human pancreatic can- cer cells and a pancreatic cancer tissue sample as judged by the retarded electrophoretic mobility of the two major FRAP downstream targets, p70s6K and 4E-BP1. Treatment of cells with rapamycin, a selective FRAP inhibitor, inhibited basal p70s6K kinase activity and induced dephospho- rylation

Martin Grewe; Frank Gansauge; Roland M. Schmid; Guido Adler; Thomas Seufferlein

1999-01-01

161

The constitution of networks  

Microsoft Academic Search

The idea of a constitution of networks may be a useful addition to our ways of understanding what happens within the business landscape. We describe a constitution of networks as a system of values, norms, rules and other conventions that are shared by actors in business networks. Whether by intention or not, the constitution of networks provides a framework within

Stefanos Mouzas; David Ford

2009-01-01

162

Isolation of SOS Constitutive Mutants of Escherichia coli†  

PubMed Central

The bacterial SOS regulon is strongly induced in response to DNA damage from exogenous agents such as UV radiation and nalidixic acid. However, certain mutants with defects in DNA replication, recombination, or repair exhibit a partially constitutive SOS response. These mutants presumably suffer frequent replication fork failure, or perhaps they have difficulty rescuing forks that failed due to endogenous sources of DNA damage. In an effort to understand more clearly the endogenous sources of DNA damage and the nature of replication fork failure and rescue, we undertook a systematic screen for Escherichia coli mutants that constitutively express the SOS regulon. We identified mutant strains with transposon insertions in 42 genes that caused increased expression from a dinD1::lacZ reporter construct. Most of these also displayed significant increases in basal levels of RecA protein, confirming an effect on the SOS system. As expected, this collection includes genes, such as lexA, dam, rep, xerCD, recG, and polA, which have previously been shown to cause an SOS constitutive phenotype when inactivated. The collection also includes 28 genes or open reading frames that were not previously identified as SOS constitutive, including dcd, ftsE, ftsX, purF, tdcE, and tynA. Further study of these SOS constitutive mutants should be useful in understanding the multiple causes of endogenous DNA damage. This study also provides a quantitative comparison of the extent of SOS expression caused by inactivation of many different genes in a common genetic background. PMID:15489426

O'Reilly, Erin K.; Kreuzer, Kenneth N.

2004-01-01

163

Isolation of SOS constitutive mutants of Escherichia coli.  

PubMed

The bacterial SOS regulon is strongly induced in response to DNA damage from exogenous agents such as UV radiation and nalidixic acid. However, certain mutants with defects in DNA replication, recombination, or repair exhibit a partially constitutive SOS response. These mutants presumably suffer frequent replication fork failure, or perhaps they have difficulty rescuing forks that failed due to endogenous sources of DNA damage. In an effort to understand more clearly the endogenous sources of DNA damage and the nature of replication fork failure and rescue, we undertook a systematic screen for Escherichia coli mutants that constitutively express the SOS regulon. We identified mutant strains with transposon insertions in 42 genes that caused increased expression from a dinD1::lacZ reporter construct. Most of these also displayed significant increases in basal levels of RecA protein, confirming an effect on the SOS system. As expected, this collection includes genes, such as lexA, dam, rep, xerCD, recG, and polA, which have previously been shown to cause an SOS constitutive phenotype when inactivated. The collection also includes 28 genes or open reading frames that were not previously identified as SOS constitutive, including dcd, ftsE, ftsX, purF, tdcE, and tynA. Further study of these SOS constitutive mutants should be useful in understanding the multiple causes of endogenous DNA damage. This study also provides a quantitative comparison of the extent of SOS expression caused by inactivation of many different genes in a common genetic background. PMID:15489426

O'Reilly, Erin K; Kreuzer, Kenneth N

2004-11-01

164

Constitutive Expression of Hypoxia-inducible Factor1 Renders Pancreatic Cancer Cells Resistant to Apoptosis Induced by Hypoxia and Nutrient Deprivation1  

Microsoft Academic Search

Hypovasculature is an outstanding characteristic of pancreatic cancers in imaging diagnosis, suggesting that blood supply is poor in pancreatic cancer tissues. Despite poor blood supply, pancreatic cancer cells survive and proliferate in severe hypoxia and nutrient deprivation. To demon- strate how pancreatic cancer cells adapt themselves to hypoxia and nu- trient deprivation, we investigated the expression of hypoxia-inducible factor 1

Nobuaki Akakura; Masanobu Kobayashi; Iori Horiuchi; Akiko Suzuki; Jingxin Wang; Jian Chen; Hiroto Niizeki; Ken-ichi Kawamura; Masuo Hosokawa; Masahiro Asaka

2001-01-01

165

Dynamic Alternations in Cellular and Molecular Components during Blossom-End Rot Development in Tomatoes Expressing sCAX1, a Constitutively Active Ca2+/H+ Antiporter from Arabidopsis1[W][OA  

PubMed Central

Although calcium (Ca) concentration in cellular compartments has been suggested to be tightly regulated, Ca deficiency disorders such as blossom-end rot (BER) in tomato (Solanum lycopersicum) fruit may be induced by abnormal regulation of Ca partitioning and distribution in the cell. The objectives of this work were to analyze the effects of high expression of the constitutively functional Arabidopsis (Arabidopsis thaliana) Ca2+/H+ exchanger (sCAX1) tonoplast protein in tomato fruit on cellular Ca partitioning and distribution, membrane integrity, and the transcriptional profile of genes potentially involved in BER development. Wild-type and sCAX1-expressing tomato plants were grown in a greenhouse. Wild-type plants did not develop BER, whereas sCAX1-expressing plants reached 100% BER incidence at 15 d after pollination. The sCAX1-expressing fruit pericarp had higher total tissue and water-soluble Ca concentrations, lower apoplastic and cytosolic Ca concentrations, higher membrane leakage, and Ca accumulation in the vacuole of sCAX1-expressing cells. Microarray analysis of healthy sCAX1-expressing fruit tissue indicated down-regulation of genes potentially involved in BER development, such as genes involved in membrane structure and repair and cytoskeleton metabolism, as well as up-regulation of genes that may have limited BER damage expansion, such as genes coding for heat shock proteins, glutathione S-transferases, and peroxidases. The results indicate that the high expression of the sCAX1 gene reduces cytosolic and apoplastic Ca concentrations, affecting plasma membrane structure and leading to BER symptom development in the fruit tissue. PMID:21464475

Tonetto de Freitas, Sergio; Padda, Malkeet; Wu, Qingyu; Park, Sunghun; Mitcham, Elizabeth J.

2011-01-01

166

A rapid and transient synthesis of nitric oxide (NO) by a constitutively expressed type II NO synthase in the guinea-pig suprachiasmatic nucleus.  

PubMed

1. We have measured extracellular NO/NO(2)(-) concentrations in guinea-pig suprachiasmatic nucleus (SCN) brain slices using fast cyclic voltammetry. A rapid and transient signal equivalent to 2.2+/-0.2 microM NO/NO(2)(-) (mean+/-s.e.mean, n=13) was detected at 1.26 V, the peak oxidation potential for NO, following local electrical stimulation (five pulses of 0.1 ms duration at 100 Hz, delivered every 5 min). 2. The NO/NO(2)(-) signal was inhibited by the non-selective nitric oxide synthase (NOS) inhibitors L-NAME, L-NMMA and the highly selective type II NOS (iNOS) inhibitor 1400 W (Garvey et al., 1997) in a concentration-dependent manner. IC(50) values were 229 microM (65 - 801, n=3, geomean and 95% confidence intervals (C.I.)), 452 nM (88 - 2310, n=5), and 14.2 microM (3.6 - 54.4, n=5), with maximum inhibitions of 82.8+/-6.7, 46.0+/-8.1, and 90.6+/-3.6%, respectively. 3. Exposure of the slices to the protein synthesis inhibitor cyclohexamide or the inhibitor of type II NOS induction dexamethasone immediately following slice cutting, and for a subsequent 4 - 5 h, did not inhibit the NO/NO(2)(-) signal. 4. The evoked NO/NO(2)(-) signal was not reduced following 6 h perfusion in Ca(2+)-free media, consistent with a Ca(2+)-independent type II NOS activity. 5. PCR for type II NOS revealed the presence of this isotype in the SCN, even immediately following removal of the brain. 6. These studies provide the first evidence to suggest a functional, constitutively-active type II NOS within the brain of normal, healthy adult animals, and add type II NOS to the multiple isotypes of NO synthase playing a role within the mammalian SCN. PMID:11682457

Starkey, S J; Grant, A L; Hagan, R M

2001-11-01

167

The Misconceived Assumption About Constitutional Assumptions  

Microsoft Academic Search

Both originalists and nonoriginalists alike often assume that background assumptions widely held when the Constitution or its amendments were enacted are part of the original meaning of the text. Originalists sometimes appeal to these background assumptions to render the meaning of more abstract words or phrases more determinate; nonoriginalist point to odious or outmoded assumptions as proof that original meaning

Randy E Barnett

2009-01-01

168

Interpreting the Constitution.  

ERIC Educational Resources Information Center

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)

Brennan, William J., Jr.

1987-01-01

169

The Constitution by Cell  

ERIC Educational Resources Information Center

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…

Greenhut, Stephanie; Jones, Megan

2010-01-01

170

Comprehensive genome-wide survey, genomic constitution and expression profiling of the NAC transcription factor family in foxtail millet (Setaria italica L.).  

PubMed

The NAC proteins represent a major plant-specific transcription factor family that has established enormously diverse roles in various plant processes. Aided by the availability of complete genomes, several members of this family have been identified in Arabidopsis, rice, soybean and poplar. However, no comprehensive investigation has been presented for the recently sequenced, naturally stress tolerant crop, Setaria italica (foxtail millet) that is famed as a model crop for bioenergy research. In this study, we identified 147 putative NAC domain-encoding genes from foxtail millet by systematic sequence analysis and physically mapped them onto nine chromosomes. Genomic organization suggested that inter-chromosomal duplications may have been responsible for expansion of this gene family in foxtail millet. Phylogenetically, they were arranged into 11 distinct sub-families (I-XI), with duplicated genes fitting into one cluster and possessing conserved motif compositions. Comparative mapping with other grass species revealed some orthologous relationships and chromosomal rearrangements including duplication, inversion and deletion of genes. The evolutionary significance as duplication and divergence of NAC genes based on their amino acid substitution rates was understood. Expression profiling against various stresses and phytohormones provides novel insights into specific and/or overlapping expression patterns of SiNAC genes, which may be responsible for functional divergence among individual members in this crop. Further, we performed structure modeling and molecular simulation of a stress-responsive protein, SiNAC128, proffering an initial framework for understanding its molecular function. Taken together, this genome-wide identification and expression profiling unlocks new avenues for systematic functional analysis of novel NAC gene family candidates which may be applied for improvising stress adaption in plants. PMID:23691254

Puranik, Swati; Sahu, Pranav Pankaj; Mandal, Sambhu Nath; B, Venkata Suresh; Parida, Swarup Kumar; Prasad, Manoj

2013-01-01

171

Comprehensive Genome-Wide Survey, Genomic Constitution and Expression Profiling of the NAC Transcription Factor Family in Foxtail Millet (Setaria italica L.)  

PubMed Central

The NAC proteins represent a major plant-specific transcription factor family that has established enormously diverse roles in various plant processes. Aided by the availability of complete genomes, several members of this family have been identified in Arabidopsis, rice, soybean and poplar. However, no comprehensive investigation has been presented for the recently sequenced, naturally stress tolerant crop, Setaria italica (foxtail millet) that is famed as a model crop for bioenergy research. In this study, we identified 147 putative NAC domain-encoding genes from foxtail millet by systematic sequence analysis and physically mapped them onto nine chromosomes. Genomic organization suggested that inter-chromosomal duplications may have been responsible for expansion of this gene family in foxtail millet. Phylogenetically, they were arranged into 11 distinct sub-families (I-XI), with duplicated genes fitting into one cluster and possessing conserved motif compositions. Comparative mapping with other grass species revealed some orthologous relationships and chromosomal rearrangements including duplication, inversion and deletion of genes. The evolutionary significance as duplication and divergence of NAC genes based on their amino acid substitution rates was understood. Expression profiling against various stresses and phytohormones provides novel insights into specific and/or overlapping expression patterns of SiNAC genes, which may be responsible for functional divergence among individual members in this crop. Further, we performed structure modeling and molecular simulation of a stress-responsive protein, SiNAC128, proffering an initial framework for understanding its molecular function. Taken together, this genome-wide identification and expression profiling unlocks new avenues for systematic functional analysis of novel NAC gene family candidates which may be applied for improvising stress adaption in plants. PMID:23691254

Puranik, Swati; Sahu, Pranav Pankaj; Mandal, Sambhu Nath; B., Venkata Suresh; Parida, Swarup Kumar; Prasad, Manoj

2013-01-01

172

Matrix Gla Protein Is a Developmental Regulator of Chondrocyte Mineralization And, When Constitutively Expressed, Blocks Endochondral and Intramembranous Ossification in the Limb  

PubMed Central

Matrix GLA protein (MGP), a ?-carboxyglutamic acid (GLA)–rich, vitamin K–dependent and apatite-binding protein, is a regulator of hypertrophic cartilage mineralization during development. However, MGP is produced by both hypertrophic and immature chondrocytes, suggesting that MGP's role in mineralization is cell stage–dependent, and that MGP may have other roles in immature cells. It is also unclear whether MGP regulates the quantity of mineral or mineral nature and quality as well. To address these issues, we determined the effects of manipulations of MGP synthesis and expression in (a) immature and hypertrophic chondrocyte cultures and (b) the chick limb bud in vivo. The two chondrocyte cultures displayed comparable levels of MGP gene expression. Yet, treatment with warfarin, a ?-carboxylase inhibitor and vitamin K antagonist, triggered mineralization in hypertrophic but not immature cultures. Warfarin effects on mineralization were highly selective, were accompanied by no appreciable changes in MGP expression, alkaline phosphatase activity, or cell number, and were counteracted by vitamin K cotreatment. Scanning electron microscopy, x-ray microanalysis, and Fourier-transform infrared spectroscopy revealed that mineral forming in control and warfarin-treated hypertrophic cell cultures was similar and represented stoichiometric apatite. Virally driven MGP overexpression in cultured chondrocytes greatly decreased mineralization. Surprisingly, MGP overexpression in the developing limb not only inhibited cartilage mineralization, but also delayed chondrocyte maturation and blocked endochondral ossification and formation of a diaphyseal intramembranous bone collar. The results show that MGP is a powerful but developmentally regulated inhibitor of cartilage mineralization, controls mineral quantity but not type, and appears to have a previously unsuspected role in regulating chondrocyte maturation and ossification processes. PMID:10579728

Yagami, Kimitoshi; Suh, Jo-Young; Enomoto-Iwamoto, Motomi; Koyama, Eiki; Abrams, William R.; Shapiro, Irving M.; Pacifici, Maurizio; Iwamoto, Masahiro

1999-01-01

173

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

PubMed

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. PMID:22316602

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

2012-04-01

174

Induction of Constitutive High-Level Expression of c-Myc in 32D Cells by Mycoplasmas is Associated with their Ability to Prevent Apoptosis and Induce Malignant Transformation.  

PubMed

Our previous studies showed that mycoplasmas prevented apoptosis and induced the malignant transformation of mammalian cells. Other studies indicate that c-Myc plays an important role in promoting apoptosis and malignant transformation of cells. To understand the role of c-Myc in the mycoplasma induced apoptosis prevention and malignant cell transformation, 32D cells, an IL-3 dependent cell line, were infected and transformed by different species of mycoplasmas. The expression of Myc and ras gene families, apoptosis and the cell cycle during the infection and transformation were examined. Results showed that c-Myc expression was significantly increased in mycoplasma transformed 32D cells. Withdrawal of IL-3 substantially decreased c-Myc expression and led to cell cycle arrest at the G1 phase followed by rapid apoptosis. Infection by M. fermentans or M. penetrans not only alleviated the sharp decrease of c-Myc expression, rescued 32D cells from cell-cycle arrest and prevented apoptosis in IL-3-free culture, but also induced autonomous growth of 32D cells. Although M. hominis and M. salivarium had the ability neither to prevent apoptosis nor to induce malignant transformation, they were still able to rescue the cells from cell cycle arrest. The expression of ras family did not change significantly during the infection and transformation. These results suggest that constitutive expression of c-Myc appears to be associated with the continuous growth and malignant transformation of 32D cells induced by M. fermentans and M. penetrans, but not with rescuing the cell cycle arrest by the mycoplasmas. PMID:23675000

Zhang, Shimin; Tsai, Shien; Lo, Shyh-Ching

2006-12-01

175

The Effect of High Concentrations of Glufosinate Ammonium on the Yield Components of Transgenic Spring Wheat (Triticum aestivum L.) Constitutively Expressing the bar Gene  

PubMed Central

We present an experiment done on a bar+ wheat line treated with 14 different concentrations of glufosinate ammonium—an effective component of nonselective herbicides—during seed germination in a closed experimental system. Yield components as number of spikes per plant, number of grains per spike, thousand kernel weight, and yield per plant were thoroughly analysed and statistically evaluated after harvesting. We found that a concentration of glufosinate ammonium 5000 times the lethal dose was not enough to inhibit the germination of transgenic plants expressing the bar gene. Extremely high concentrations of glufosinate ammonium caused a bushy phenotype, significantly lower numbers of grains per spike, and thousand kernel weights. Concerning the productivity, we observed that concentrations of glufosinate ammonium 64 times the lethal dose did not lead to yield depression. Our results draw attention to the possibilities implied in the transgenic approaches. PMID:22649303

Áy, Zoltán; Mihály, Róbert; Cserháti, Mátyás; Kótai, Éva; Pauk, János

2012-01-01

176

Constitutive Expression of a miR319 Gene Alters Plant Development and Enhances Salt and Drought Tolerance in Transgenic Creeping Bentgrass1[W][OA  

PubMed Central

MicroRNA319 (miR319) is one of the first characterized and conserved microRNA families in plants and has been demonstrated to target TCP (for TEOSINTE BRANCHED/CYCLOIDEA/PROLIFERATING CELL FACTORS [PCF]) genes encoding plant-specific transcription factors. MiR319 expression is regulated by environmental stimuli, suggesting its involvement in plant stress response, although experimental evidence is lacking and the underlying mechanism remains elusive. This study investigates the role that miR319 plays in the plant response to abiotic stress using transgenic creeping bentgrass (Agrostis stolonifera) overexpressing a rice (Oryza sativa) miR319 gene, Osa-miR319a. We found that transgenic plants overexpressing Osa-miR319a displayed morphological changes and exhibited enhanced drought and salt tolerance associated with increased leaf wax content and water retention but reduced sodium uptake. Gene expression analysis indicated that at least four putative miR319 target genes, AsPCF5, AsPCF6, AsPCF8, and AsTCP14, and a homolog of the rice NAC domain gene AsNAC60 were down-regulated in transgenic plants. Our results demonstrate that miR319 controls plant responses to drought and salinity stress. The enhanced abiotic stress tolerance in transgenic plants is related to significant down-regulation of miR319 target genes, implying their potential for use in the development of novel molecular strategies to genetically engineer crop species for enhanced resistance to environmental stress. PMID:23292790

Zhou, Man; Li, Dayong; Li, Zhigang; Hu, Qian; Yang, Chunhua; Zhu, Lihuang; Luo, Hong

2013-01-01

177

An integrative expression vector for Actinosynnema pretiosum  

Microsoft Academic Search

BACKGROUND: The Actinomycete Actinosynnema pretiosum ssp. auranticum has commercial importance due to its production of ansamitocin P-3 (AP-3), a potent antitumor agent. One way to increase AP-3 production would be to constitutively express selected genes so as to relieve bottlenecks in the biosynthetic pathway; however, an integrative expression vector for A. pretiosum is lacking. The aim of this study was

Shan Goh; Andrea Camattari; Daniel Ng; Ruth Song; Kevin Madden; Janet Westpheling; Victor VT Wong

2007-01-01

178

Thermomechanical constitutive model of shape memory polymer  

Microsoft Academic Search

A nonlinear thermomechanical constitutive model of shape memory polymer (SMP) is developed by modifying a linear model. The coefficients in the model are expressed by the single exponential functions of temperature in order to describe the variation in mechanical properties of the material due to the glass transition. The proposed theory expresses well the thermomechanical properties of polyurethane-shape memory polymer,

Hisaaki Tobushi; Kayo Okumura; Shunichi Hayashi; Norimitsu Ito

2001-01-01

179

Constitutive Expression of the Proteorhodopsin Gene by a Flavobacterium Strain Representative of the Proteorhodopsin-Producing Microbial Community in the North Sea? †  

PubMed Central

Proteorhodopsin (PR), a photoactive proton pump containing retinal, is present in approximately half of all bacteria in the ocean, but its physiological role is still unclear, since very few strains carrying the PR gene have been cultured. The aim of this work was to characterize PR diversity in a North Sea water sample, cultivate a strain representative of North Sea PR clusters, and study the effects of light and carbon concentration on the expression of the PR gene. A total of 117 PR sequences, of which 101 were unique, were obtained from a clone library of PCR-amplified PR gene fragments. Of the North Sea PRs, 97% were green light absorbing, as inferred from the amino acid at position 105; 67% of the PR protein fragments showed closest similarity to PRs from Alphaproteobacteria, 4% showed closest similarity to PRs from Gammaproteobacteria, and 29% showed closest similarity to PRs from “Bacteroidetes”/Flavobacteria. The dominant PR cluster (comprising 18% of all PRs) showed a high degree of similarity to the PR from the cultivated Roseobacter strain HTCC2255. The relative abundances of the North Sea PR clusters were confirmed by quantitative PCR. They were detected in metagenomic fragments from coastal oceans worldwide with various degrees of abundance. Several hundred bacterial strains from the North Sea water sample were cultivated on oligocarbophilic media. By screening with degenerate primers, two strains carrying the PR gene were identified. Their 16S rRNA gene sequences were identical and affiliated with a Bacteroidetes subcluster from the North Sea. The PR sequence of isolate PRO95 was completed by chromosomal walking. It was 76% identical to that of Dokdonia donghaensis MED134 and was functional, as indicated by the signature amino acids. PRO95 expressed its PR gene in liquid media containing between 9.7 and 121 mM carbon, both in the light and in the dark. Growth was not enhanced by light. Thus, the detection of the physiological role of PR may require more sensitive methods. PMID:20305030

Riedel, Thomas; Tomasch, Jürgen; Buchholz, Ina; Jacobs, Jenny; Kollenberg, Mario; Gerdts, Gunnar; Wichels, Antje; Brinkhoff, Thorsten; Cypionka, Heribert; Wagner-Döbler, Irene

2010-01-01

180

The Trypanosoma brucei DNA polymerase alpha core subunit gene is developmentally regulated and linked to a constitutively expressed open reading frame.  

PubMed Central

As an initial step towards the characterization of replicative DNA polymerases of trypanosomes, we have cloned, sequenced and examined the expression of the Trypanosoma (Trypanozoon) brucei brucei gene that encodes the DNA polymerase alpha catalytic core (pol alpha). The protein sequence contains the six conserved regions that have been recognized previously in eukaryotic and viral replicative DNA polymerases. In addition, we have identified a seventh region which appears to be conserved primarily in alpha-type DNA polymerases. The T.brucei DNA pol alpha core N-terminus is 123 and 129 amino acids smaller than that of the human and yeast homologue, respectively. The gene is separated by 386 bp from an upstream open reading frame (ORF) of 442 codons. Stable transcripts of the upstream sequence are detected in both dividing and non-dividing forms, while pol alpha transcripts are detected principally in dividing forms. Allelic copies of the T.brucei pol alpha region exhibit restriction site polymorphisms; one such sequence polymorphism affects the amino acid sequence of the T.brucei DNA pol alpha core. The T.brucei pol alpha region cross-hybridizes weakly with that of T.(Nannomonas) congolense and T.(Duttonella) vivax. Images PMID:1754381

Leegwater, P A; Strating, M; Murphy, N B; Kooy, R F; van der Vliet, P C; Overdulve, J P

1991-01-01

181

The growth of simian virus 40 (SV40) host range/adenovirus helper function mutants in an African green monkey cell line that constitutively expresses the SV40 agnoprotein.  

PubMed Central

The simian virus 40 T-antigen carboxy-terminal mutants, dlA2459 and dlA2475, are cell line and temperature dependent for growth and plaque formation in monkey kidney cells. Although these mutants did form plaques on BSC-1 cells at 37 degrees C, they were about fivefold less efficient for plaque formation than wild-type simian virus 40. These mutants did not grow in CV-1 cells and did not synthesize agnoprotein in those cells. CV-1 cells which constitutively express the agnoprotein were permissive for mutant plaque formation. However, late mRNAs, virion proteins, and progeny virion yields did not accumulate to wild-type levels during mutant infection of the agnoprotein-producing cells. Images PMID:2161954

Stacy, T P; Chamberlain, M; Carswell, S; Cole, C N

1990-01-01

182

Classroom Constitutional Conventions.  

ERIC Educational Resources Information Center

This lesson provides a simulation in which upper elementary and middle school students establish classroom routines, participate in decision-making activities, improve their understanding of the Constitutional Convention, and differentiate among the three branches of government. (BSR)

Roach, Carol

1987-01-01

183

Kenya's New Constitution  

E-print Network

On 4 August 2010, Kenyans voted to adopt a new constitution, culminating a process that began as part of a resolution to the violent conflict that followed the December 2007 elections. By reducing executive power, devolving ...

Kramon, Eric

184

Constitution and Bylaws.  

ERIC Educational Resources Information Center

Constitutions and bylaws of the American Association of Dental Schools are presented, covering organization name, statement of purpose, administrative organization, offices, membership, legislative and administrative bodies, elected and appointed officers, regular and special sessions, sections, publications, election, and finance. (MSE)

Journal of Dental Education, 1981

1981-01-01

185

HYPOPROLIFERATIVE Acquired or Constitutional  

E-print Network

9/16/2013 1 HYPOPROLIFERATIVE ANEMIAS Acquired or Constitutional Disorder with Chronic Bone Marrow, or megakaryocytic hypoplasia used APLASTIC ANEMIA Criteria Bone Cellularity : Granulocyte Count Anemia with corrected Retic Count

186

Purification from rat liver of a novel constitutively expressed member of the aldo-keto reductase 7 family that is widely distributed in extrahepatic tissues.  

PubMed

Antiserum raised against human aflatoxin B(1) aldehyde reductase 1 (hAFAR1) has been used to identify a previously unrecognized rat aldo-keto reductase (AKR). This novel enzyme is designated rat aflatoxin B(1) aldehyde reductase 2 (rAFAR2) and it characteristically migrates faster during SDS/PAGE than does the archetypal ethoxyquin-inducible rAFAR protein (now called rAFAR1). Significantly, rAFAR2 is essentially unreactive with polyclonal antibodies raised against rAFAR1. Besides its distinct electrophoretic and immunochemical properties, rAFAR2 appears to be regulated differently from rAFAR1 as it is expressed in most rat tissues and does not appear to be induced by ethoxyquin. Multiple forms of rAFAR2 have been identified. Anion-exchange chromatography on Q-Sepharose, followed by adsorption chromatography on columns of Matrex Orange A and Cibacron Blue, have been employed to purify rAFAR2 from rat liver cytosol. The Q-Sepharose chromatography step resulted in the resolution of rAFAR2 into three peaks of AKR activity, two of which were purified and shown to be capable of catalysing the reduction of 2-carboxybenzaldehyde, succinic semialdehyde, 4-nitrobenzaldehyde and 9,10-phenathrenequinone. The two most highly purified rAFAR2-containing preparations eluted from the Cibacron Blue column were 91 and 98% homogeneous. Analysis of these by SDS/PAGE indicated that the least anionic (peak CBA5) comprised a polypeptide of 37.0 kDa, whereas the most anionic (peak CBA6) contained two closely migrating polypeptides of 36.8 and 37.0 kDa; by contrast, in the present study, rAFAR1 was estimated by SDS/PAGE to be composed of 38.0 kDa subunits. Final purification of the 37 kDa polypeptide in CBA5 and CBA6 was accomplished by reversed-phase HPLC. Partial proteolysis of the two preparations of the 37 kDa polypeptide with Staphylococcus aureus V8 protease yielded fragments of identical size, suggesting that they represent the product of a single gene. Furthermore, the peptide maps from CBA5 and CBA6 differed substantially from that yielded by rAFAR1, indicating that they are genetically distinct from the inducible reductase. A peptide generated by CNBr digestion of the 37 kDa polypeptide from CBA6 was shown by Edman degradation to share 88% sequence identity with residues Tyr(168)-Leu(183) of rAFAR1. This provides evidence that the rat protein identified by its cross-reactivity with anti-hAFAR1 serum is an additional member of the AKR7 family. PMID:10816434

Kelly, V P; Ireland, L S; Ellis, E M; Hayes, J D

2000-06-01

187

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

PubMed

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 pGAPZ?A-LaccGluc-Stop and pGAPZ?A-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 ?-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 that laccases can transform. This contributes to a great gamut of products in diverse settings: industry, clinical and chemical use, and environmental applications. PMID:25611746

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

2015-01-01

188

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

PubMed Central

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 pGAPZ?A-LaccGluc-Stop and pGAPZ?A-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 ?-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 that laccases can transform. This contributes to a great gamut of products in diverse settings: industry, clinical and chemical use, and environmental applications. PMID:25611746

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

2015-01-01

189

Human & Constitutional Rights  

NSDL National Science Digital Library

The Arthur W. Diamond Law Library at Columbia Law School maintains this excellent resource for finding materials on human rights and constitutional rights. The metasite serves students, scholars, and practitioners as a portal to documents and Internet resources on international and domestic law related to human and constitutional rights. The information resources are divided into six sections: Country Reports, International Links, Regional Links, National Links, Documents, and Other Web Resources. Each section is clearly organized into neat lists or pop-up menus to ease navigation. Marylin Raisch -- the International, Comparative, and Foreign Law Librarian responsible for this metasite -- also provides a Hot Topics section, which posts information on current events related to human and constitutional rights.

190

Crushed Salt Constitutive Model  

SciTech Connect

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.

Callahan, G.D.

1999-02-01

191

The Constitution in Action  

ERIC Educational Resources Information Center

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…

Potter, Lee Ann

2007-01-01

192

Gut microbiota, tight junction protein expression, intestinal resistance, bacterial translocation and mortality following cholestasis depend on the genetic background of the host.  

PubMed

Failure of the intestinal barrier is a characteristic feature of cholestasis. We have previously observed higher mortality in C57BL/6J compared with A/J mice following common bile duct ligation (CBDL). We hypothesized the alteration in gut barrier function following cholestasis would vary by genetic background. Following one week of CBDL, jejunal TEER was significantly reduced in each ligated mouse compared with their sham counterparts; moreover, jejunal TEER was significantly lower in both sham and ligated C57BL/6J compared with sham and ligated A/J mice, respectively. Bacterial translocation to mesenteric lymph nodes was significantly increased in C57BL/6J mice vs. A/J mice. Four of 15 C57BL/6J mice were bacteremic; whereas, none of the 17 A/J mice were. Jejunal IFN-? mRNA expression was significantly elevated in C57BL/6J compared with A/J mice. Western blot analysis demonstrated a significant decrease in occludin protein expression in C57BL/6J compared with A/J mice following both sham operation and CBDL. Only C57BL/6J mice demonstrated a marked decrease in ZO-1 protein expression following CBDL compared with shams. Pyrosequencing of the 16S rRNA gene in fecal samples showed a dysbiosis only in C57BL/6J mice following CBDL when compared with shams. This study provides evidence of strain differences in gut microbiota, tight junction protein expression, intestinal resistance and bacterial translocation which supports the notion of a genetic predisposition to exaggerated injury following cholestasis. PMID:23652772

Alaish, Samuel M; Smith, Alexis D; Timmons, Jennifer; Greenspon, Jose; Eyvazzadeh, Daniel; Murphy, Ebony; Shea-Donahue, Terez; Cirimotich, Shana; Mongodin, Emmanuel; Zhao, Aiping; Fasano, Alessio; Nataro, James P; Cross, Alan

2013-01-01

193

Gut microbiota, tight junction protein expression, intestinal resistance, bacterial translocation and mortality following cholestasis depend on the genetic background of the host  

PubMed Central

Failure of the intestinal barrier is a characteristic feature of cholestasis. We have previously observed higher mortality in C57BL/6J compared with A/J mice following common bile duct ligation (CBDL). We hypothesized the alteration in gut barrier function following cholestasis would vary by genetic background. Following one week of CBDL, jejunal TEER was significantly reduced in each ligated mouse compared with their sham counterparts; moreover, jejunal TEER was significantly lower in both sham and ligated C57BL/6J compared with sham and ligated A/J mice, respectively. Bacterial translocation to mesenteric lymph nodes was significantly increased in C57BL/6J mice vs. A/J mice. Four of 15 C57BL/6J mice were bacteremic; whereas, none of the 17 A/J mice were. Jejunal IFN-? mRNA expression was significantly elevated in C57BL/6J compared with A/J mice. Western blot analysis demonstrated a significant decrease in occludin protein expression in C57BL/6J compared with A/J mice following both sham operation and CBDL. Only C57BL/6J mice demonstrated a marked decrease in ZO-1 protein expression following CBDL compared with shams. Pyrosequencing of the 16S rRNA gene in fecal samples showed a dysbiosis only in C57BL/6J mice following CBDL when compared with shams. This study provides evidence of strain differences in gut microbiota, tight junction protein expression, intestinal resistance and bacterial translocation which supports the notion of a genetic predisposition to exaggerated injury following cholestasis. PMID:23652772

Alaish, Samuel M.; Smith, Alexis D.; Timmons, Jennifer; Greenspon, Jose; Eyvazzadeh, Daniel; Murphy, Ebony; Shea-Donahue, Terez; Cirimotich, Shana; Mongodin, Emmanuel; Zhao, Aiping; Fasano, Alessio; Nataro, James P.; Cross, Alan S

2013-01-01

194

Explore the Constitution  

NSDL National Science Digital Library

If you type the words â??U.S. Constitutionâ?ť into Google, the search engine returns about 31 million results. Thatâ??s a lot of talk about a document written by a handful of scholar-politicians in an upstart republic more than 227 years ago. Yet less than a quarter of Americans have actually read the document. This site, hosted by the National Constitution Center, includes the United States Constitution in its entirety, divided clearly by Article and Amendment, and annotated with notes by the Annenberg Classroom. Perhaps best of all, you can browse the site by Issue. For instance, click on Abortion and explore how the Roe v. Wade Supreme Court ruling was based on Ninth and 14th Amendment rights.

195

Nepal's Constitutional Process  

E-print Network

to the difficult questions of procedure involved in constitutional reform. Few have embarked on internal changes to tackle their own problems of corruption, patronage and exclusion that fuelled support for the Maoists. Strengthening parties’ internal democracy... will be permitted without interference or violence; (b) allowing the police, as agreed in the CPA, to maintain order and investigate criminal activities; and (c) committing publicly to respect freedoms of speech and political association of other political...

International Crisis Group

2007-02-26

196

Thomas Jefferson and the Constitution.  

ERIC Educational Resources Information Center

Examines Thomas Jefferson's role in the making and interpretation of the United States Constitution. Discusses the dominant features of Jefferson's constitutional theory; the character of Jefferson's presidency; and Jefferson's ongoing concern about constitutional preservation and change. Lists important dates in the history of the constitution.…

Peterson, Merrill D.

1987-01-01

197

Conscience and the Constitution  

NSDL National Science Digital Library

This companion site to the PBS national broadcast December 2000, of "Conscience and the Constitution" examines the historical and moral issues surrounding the decision of a handful of Japanese to refuse to be drafted from their location in an American concentration camp in 1944. The site also looks at the history of the Japanese American Citizens League (JACL), an organization of first generation Japanese-Americans that not only supported the efforts of the US military to apprehend and relocate Japanese citizens, but opposed all efforts of individual Japanese-Americans to challenge the constitutionality of the government's actions. In addition to the thoughtful narration of the history and the controversy it continues to generate, the Website provides access to original documents of the parties involved, including letters and statements from the draft resisters, who eventually served more than two years hard labor, the columns of the journalist who supported their civil disobedience, and public statements and correspondence of the JACL. Some video portions of the broadcast are also available here in RealPlayer.

198

Ethics and constitutional government.  

PubMed

The term ethics refers to a set of principles that govern acceptable, proper conduct. Attacks on the Constitution of the United States pose the most serious breach of ethics today. Our country was founded as a republic, not as a democracy. Our Founding Fathers' main concern was to protect citizens from the power of the federal government, so constitutionally, the central government has little or no authority over individual citizens except on federal property. One of the major problems today is the fact that we now have professional politicians. This is due in large part to the lure of financial gain from countless special interest groups. This would change under constitutional law because the federal budget would decrease drastically. Article 1 states that all legislative power is vested in Congress. Congress has only 18 enumerated powers, and almost half of these pertain to defense of the country. Many of our current problems are due to regulatory agencies that have become independent fiefdoms with unconstitutional legislative, as well as executive and judicial, powers. The regulatory agency most relevant to medicine, both clinical care and research, is the FDA. It is now obvious that its basic structure needs to be changed or abolished because its actions are identical to those inherent in authoritarian systems. Constructive change could come from Congress, but it would be most desirable if the Supreme Court would take the lead and reestablish the authority of the Constitution as the Supreme Law of the Land. The FDA's function could be limited to the determination of safety, but preferably its mission would be altered to that of product certification. Defenders of the current system claim that such a drastic change would be too dangerous and their prime example is thalidomide. But it is now known that the market has already solved that problem prior to the government-imposed sanctions. Realistically, market forces and their ramifications, including our legal system, provide the most effective methods of protecting the public from harmful drugs and devices. Fortunately, a model for miracles is available. It is New Zealand, which had become increasingly socialistic after WW II. As a result, they had become noncompetitive, and with the formation of the European Union, they lost their major market exports. In order to survive they made some astounding changes in the mid-1980s. They studied every agency that depended on government funding and transferred much of this work over to the private sector. They turned the remaining agencies into profit-making enterprises. These agencies had cost the government about $1 billion/year. Now, they produce about $1 billion in revenue and taxes. Without question, a return to constitutional government would be an invigorating stimulus to bioengineering research in the future. It would flourish. The eminence of this country did not develop from a strong central government. It is due to its absence, but we are rapidly reaching a point of no return. PMID:18298395

Albright, James A

2007-01-01

199

Polity.org: Constitutional Matters  

NSDL National Science Digital Library

Full text of the new South African Constitution, adopted by the Constitutional Assembly on May 8, 1996, and the full text of President Nelson Mandela's address to the Constitutional Assembly upon its adoption, are available via the Web. The constitution is available as a single file, and as plain text, WordPerfect 5.1, 6.1 and Microsoft Word 6.0 .zip files.

1996-01-01

200

STELLENBOSCH UNIVERSITY STUDENT PARLIAMENT CONSTITUTION  

E-print Network

in conformity with the Bill of Student Rights in chapter two (2) of the Student Constitution, the principles part of this constitution, or the Student Constitution, explicitly states a later date of application country, the Republic of South Africa, unite to build a multicultural, inclusive and democratic community

Geldenhuys, Jaco

201

Constitutive models in LAME.  

SciTech Connect

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 and the methods necessary for achieving accurate and efficient solutions have been incorporated. The most important method is the getStress function where the actual material model evaluation takes place. Obviously, all material models incorporate this function. The initialize function is included in most material models. The initialize function is called once at the beginning of an analysis and its primary purpose is to initialize the material state variables associated with the model. Many times, there is some information which can be set once per load step. For instance, we may have temperature dependent material properties in an analysis where temperature is prescribed. Instead of setting those parameters at each iteration in a time step, it is much more efficient to set them once per time step at the beginning of the step. These types of load step initializations are performed in the loadStepInit method. The final function used by many models is the pcElasticModuli method which changes the moduli that are to be used by the elastic preconditioner in Adagio. The moduli for the elastic preconditioner are set during the initialization of Adagio. Sometimes, better convergence can be achieved by changing these moduli for the elastic preconditioner. For instance, it typically helps to modify the preconditioner when the material model has temperature dependent moduli. For many material models, it is not necessary to change the values of the moduli that are set initially in the code. Hence, those models do not have pcElasticModuli functions. All four of these methods receive information from the matParams structure as described by Scherzinger and Hammerand.

Hammerand, Daniel Carl; Scherzinger, William Mark

2007-09-01

202

DISROBED: THE CONSTITUTION OF MODESlY ANITA L. ALLEN  

E-print Network

of the United States Constitution protects free- dom of speech and expression.l The Fourteenth Amendment pro2006] DISROBED: THE CONSTITUTION OF MODESlY ANITA L. ALLEN 1. INTRODUCTION THE First Amendment to Western Europe, topless sunbathing is rarely permitted in the United States. Even infant breast

Sharp, Kim

203

Guidelines for Teaching and Testing the Illinois Constitution.  

ERIC Educational Resources Information Center

This document sets forth guidelines for teaching and testing public school students on the principles of democratic government as expressed in the Constitution of the United States and the Constitution of the State of Illinois. Identification is made of ways of making the examination required by the state of Illinois not only an integral part of…

Illinois State Office of the Superintendent of Public Instruction, Springfield. Instructional Services Unit.

204

Conversations on the Constitution  

NSDL National Science Digital Library

Perhaps the United States might be a better place if at the dinner table one interested party might ask the question, "Hey Mom, can we talk about the establishment clause tonight?" Until such topics spontaneously arrive in a pre-prandial setting like the one just mentioned, there will still be great interest in this lovely website created by the American Bar Association. As its mission statement declares, "This project is designed to further dialogue in schools and in the workplace about American constitutional principles and values." Within the site, visitors can peruse a list of topics, such as war powers, the establishment of religion, and cruel and unusual punishment. Each area has a brief overview of each topic, and then three "starter" questions. These starter questions feature a list of specific queries, accompanied by brief illustrations (such as political cartoons and the like) that can be used at the dinner table, the boardroom table, or just about any table. Finally, interested parties can also take a few interactive tests on such areas as Supreme Court rulings and students' rights in the classroom.

205

The Constitution Project  

NSDL National Science Digital Library

Recently, The Constitution Project (TCP) started the bipartisan Task Force on Detainee Treatment. The purpose of TCP is to "bring the American people a comprehensive understanding of what is known and what may still be unknown about the past and current treatment of detainees by the U.S. government, as part of the counterterrorism policies of the Obama, Bush, and Clinton administrations." Visitors to their site can learn about their efforts to shed light on these various activities, and it is a good idea to take a quick look over their "Headline News" area to get started. Users should not miss the report "Principles for Government Data Mining", which can be found near the bottom of the homepage. This section is dedicated to looking into the threats to civil liberties posed by government data mining programs and recommends a series of reforms. Along with reading the report here, users can watch a webcast that features a panel of experts discussing the report's findings and future directions.

206

The Texas Constitutions Digitization Project  

NSDL National Science Digital Library

Presented by the Tarlton Law Library at the University of Texas School of law, this site offers digital copies of the twelve constitutions of the State (and Republic) of Texas drafted between 1824 and 1876. Both transcriptions and digital images of the original documents are provided, along with a bibliography of printings, a supplemental bibliography, and a link to the relevant Handbook of Texas article. A Spanish version is also available for a few of the constitutions. An internal search engine, an annotated bibliography on the constitutions, a bibliography on Texas constitutional history, and a collection of related links are also provided.

207

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

ERIC Educational Resources Information Center

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…

National Archives and Records Administration, Washington, DC.

208

Expression of macrophage migration inhibitory factor in human glomerulonephritis  

Microsoft Academic Search

Expression of macrophage migration inhibitory factor in human glomerulonephritis.BackgroundWe have recently demonstrated that macrophage migration inhibitory factor (MIF) plays a pathogenic role in experimental glomerulonephritis (GN). The aim of the current study was to investigate MIF expression in human GN.MethodsMIF expression was examined by in situ hybridization and immunohistochemistry staining in 65 biopsies from a variety of glomerulonephridities.ResultsThere is constitutive

Hui Y Lan; Niansheng Yang; David J Nikolic-Paterson; Xue Q Yu; Wei Mu; Nicole M Isbel; Christine N Metz; Richard Bucala; Robert C Atkins

2000-01-01

209

Background Notes  

NSDL National Science Digital Library

Country Background Notes, distributed on the DOSBACK list, are updated periodically and include information on US bilateral relations with foreign countries and on their governments, political conditions, and foreign relations. You can expect the DOSBACK list to generate about 3-4 email messages per month. Via DOSBACK you will receive the full-text version of newly released Background Notes. Archives of these two lists are also available at the Department of State Foreign Affairs Network (DOSFAN) gopher at the University of Illinois-Chicago.

210

Formal State Constitutional Change Processesi  

Microsoft Academic Search

Constitutional change in democracies occurs in two ways: by altering the meaning of the document through interpretation, or by altering the text of the document through amendment or revision. For the United States Constitution, change through interpretation predominates. Review of the extensive experience of the American states with \\

Gerald Benjamin

211

CONSTITUTIONAL LAW Political Science 320  

E-print Network

@cofc.edu ________________________________________________________________________ I. CONTENT & OBJECTIVES This course is an examination of the United States Constitution1 CONSTITUTIONAL LAW Political Science 320 Maybank Room 111 T/TH 10:50-12:05 Spring 2013 branches of the federal government, the balance of power between the state and federal government

Young, Paul Thomas

212

The Constitution and Its Critics  

ERIC Educational Resources Information Center

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…

Main, Thomas J.

2011-01-01

213

CONSTITUTION OF THE GRADUATE SCHOOL  

E-print Network

CONSTITUTION OF THE GRADUATE SCHOOL April 27, 1970 As Amended January 1971 As Amended June 1976 November 2007 As Amended September 2008 #12;CONSTITUTION THE GRADUATE SCHOOL OF THE UNIVERSITY OF MAINE ARTICLE I -- Name and Purpose. Section 1. Name. This organization shall be known as the Graduate School

Thomas, Andrew

214

Background & Publications  

E-print Network

Home Background & Projects Calendar Publications Staff Directory Station Videos Links Search Northern Michigan FruitNet 2008 Weekly Update NW Michigan Horticultural Research Station Nikki Rothwell maggot from growers. European red mites and two-spotted spider mites have reached threshold levels

215

Constitutional aneuploidy and cancer predisposition†  

PubMed Central

Constitutional aneuploidies are rare syndromes associated with multiple developmental abnormalities and the alterations in the risk for specific cancers. Acquired somatic chromosomal aneuploidies are the most common genetic aberrations in sporadic cancers. Thus studies of these rare constitutional aneuploidy syndromes are important not only for patient counseling and clinical management, but also for deciphering the mechanisms by which chromosomal aneuploidy affect cancer initiation and progression. Here we review the major constitutional aneuploidy syndromes and suggest some general mechanisms for the associated cancer predisposition. PMID:19297405

Ganmore, Ithamar; Smooha, Gil; Izraeli, Shai

2009-01-01

216

Remarks on turbulent constitutive relations  

NASA Technical Reports Server (NTRS)

The paper demonstrates that the concept of turbulent constitutive relations can be used to construct general models for various turbulent correlations. Some of the Generalized Cayley-Hamilton formulas for relating tensor products of higher extension to tensor products of lower extension are introduced. The combination of dimensional analysis and invariant theory can lead to 'turbulent constitutive relations' (or general turbulence models) for, in principle, any turbulent correlations. As examples, the constitutive relations for Reynolds stresses and scalar fluxes are derived. The results are consistent with ones from Renormalization Group (RNG) theory and two-scale Direct-Interaction Approximation (DIA) method, but with a more general form.

Shih, Tsan-Hsing; Lumley, John L.

1993-01-01

217

A role for Toll-like receptors in acquired immunity: up-regulation of TLR9 by BCR triggering in naive B cells and constitutive expression in memory B cells  

Microsoft Academic Search

Toll-like receptors (TLRs) are pattern rec- ognition receptors that trigger innate im- munity. In this study we investigated the expression of 10 TLRs in human naive and memory B-cell subsets. We report that in human naive B cells most TLRs are expressed at low to undetectable levels, but the expression of TLR9 and TLR10 is rapidly induced following B-cell-receptor (BCR)

Nadia L. Bernasconi; Nobuyuki Onai; Antonio Lanzavecchia

218

Abnormal human sex chromosome constitutions  

SciTech Connect

Chapter 22, discusses abnormal human sex chromosome constitution. Aneuploidy of X chromosomes with a female phenotype, sex chromosome aneuploidy with a male phenotype, and various abnormalities in X chromosome behavior are described. 31 refs., 2 figs.

NONE

1993-12-31

219

WE THE PEOPLE: THE CITIZEN AND THE CONSTITUTION QUESTIONS FOR SIMULATED LEGISLATIVE HEARINGS  

E-print Network

the United States Constitution and the purposes of government. The members of your group are expert witnesses did the compromise of later adding a Bill of Rights affect our Constitution? Do you agree? #12;UNIT FOUR: HOW DOES THE CONSTITUTION PROTECT OUR BASIC RIGHTS? What is freedom of expression

Garfunkel, Eric

220

Phenotypic and histological expression of different genetic backgrounds in interactions between lettuce, wild Lactuca spp., L. sativa × L. serriola hybrids and Bremia lactucae  

Microsoft Academic Search

Phenotypic and histological responses of cultivated lettuce (Lactuca sativa) and wild relatives L. saligna, L.?virosa as well as interspecific crosses derived from L. sativa × L. serriola to two races of Bremia lactucae (CS2, CS9) were investigated. With the exception of L. sativa genotypes, all accessions and hybrids expressed incomplete or complete resistance to both pathogen races, with slight differences

Aleš Lebeda; Michaela Sedlá?ová; James Lynn; David A. C. Pink

2006-01-01

221

Aminocyclopropane Carboxylic Acid Synthase Is a Regulated Step in Ethylene-Dependent Induced Conifer Defense. Full-Length cDNA Cloning of a Multigene Family, Differential Constitutive, and Wound- and Insect-Induced Expression, and Cellular and Subcellular Localization in Spruce and Douglas Fir12[W][OA  

PubMed Central

In conifer stems, formation of chemical defenses against insects or pathogens involves specialized anatomical structures of the phloem and xylem. Oleoresin terpenoids are formed in resin duct epithelial cells and phenolics accumulate in polyphenolic parenchyma cells. Ethylene signaling has been implicated in the induction of these chemical defenses. Recently, we reported the cloning of 1-aminocyclopropane-1-carboxylic acid oxidase (ACO) from spruce (Picea spp.) and Douglas fir (Pseudotsuga menziesii). ACO protein was constitutively expressed in Douglas fir and only weakly induced upon wounding. We now cloned seven full-length and one near full-length cDNA representing four distinct 1-aminocyclopropane-1-carboxylic acid synthases (ACS; ACS1, ACS2, ACS3, and ACS4) from spruce and Douglas fir. Cloning of ACS has not previously been reported for any gymnosperm. Using gene-specific, quantitative real-time polymerase chain reaction, we measured constitutive expression for the four ACS genes and the single-copy ACO gene in various tissues of Sitka spruce (Picea sitchensis) and in white spruce (Picea glauca) somatic embryos. ACO and ACS4 were ubiquitously expressed at high levels; ACS1 was predominantly expressed in developing embryos and ACS2 and ACS3 were expressed only at very low levels. Insect attack or mechanical wounding caused strong induction of ACS2 and ACS3 in Sitka spruce bark, a moderate increase in ACO transcripts, but had no effect on ACS1 and ACS4. ACS protein was also strongly induced following mechanical wounding in Douglas fir and was highly abundant in resin duct epithelial cells and polyphenolic parenchyma cells. These results suggest that ACS, but not ACO, is a regulated step in ethylene-induced conifer defense. PMID:17122070

Ralph, Steven G.; Hudgins, J.W.; Jancsik, Sharon; Franceschi, Vincent R.; Bohlmann, Jörg

2007-01-01

222

Liberty Library of Constitutional Classics  

NSDL National Science Digital Library

The Liberty Library of Constitutional Classics is a treasure trove of political primary documents that have influenced the U.S. government as well as many other countries with Republican forms of government around the world. The site makes available documents online it a variety of formats, each format is given a symbol the user can then choose which format (html, pdf, word, etc) they would prefer. The primary documents range from the Code of Hammurabi (c.1780 BCE), to Civil Disobedience by Henry David Thoreau (1849). The documents are arranged chronologically to make searching simple. Students will find this site especially useful and perhaps even a cost effective way of accessing documents for class readings or research. Instructors will also enjoy the easy access to these sources. The site also contains links to other collections including, U.S. State Constitutions, National Constitutions (from around the world), and Landmark Court Decisions.

Roland, Jon

2006-11-23

223

President Mandela Address to Constitutional Assembly  

NSDL National Science Digital Library

Full text of the new South African Constitution, adopted by the Constitutional Assembly on May 8, 1996, and the full text of President Nelson Mandela's address to the Constitutional Assembly upon its adoption, are available via the Web.

Mandela, Nelson, 1918-.

1996-01-01

224

17 CFR 200.54 - Constitutional obligations.  

Code of Federal Regulations, 2010 CFR

...2010-04-01 2010-04-01 false Constitutional obligations. 200.54 Section... Canons of Ethics § 200.54 Constitutional obligations. The members...guard against any infringement of the constitutional rights, privileges, or...

2010-04-01

225

Take Advantage of Constitution Day  

ERIC Educational Resources Information Center

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 idea and…

McCune, Bonnie F.

2008-01-01

226

The Constitution: A Resource Guide.  

ERIC Educational Resources Information Center

Presents a 234-item annotated bibliography of a wide variety of resources for teaching about the Constitution at all grade levels. The materials include: books; computer software; inspirational literature; teaching aids such as letters, memoirs, and diaries; teacher's handbooks; portraits and pictures; music, songbooks, and recordings; and…

Social Studies Review, 1987

1987-01-01

227

EUROPE'S CONSTITUTIONAL IMBROGLIO. Grard Roland  

E-print Network

in order to figure out how to go forward. What does this rejection spell for the future of the European later on June 1 in the Netherlands (61.6%) delivered a painful end to a constitutional process that had task. The IGC that took place in the fall of 2003 could not make any progress over the work

Sadoulet, Elisabeth

228

Civil Liberties under State Constitutions.  

ERIC Educational Resources Information Center

Discusses "new judicial federalism" as the rediscovery of state bills of rights for protecting civil liberties. Analyzes the differences in protection under state and federal bills of rights. Recommends that the growing importance of state constitutions be included in the political science curriculum and suggests bibliographic sources to be used.…

Tarr, G. Alan

1988-01-01

229

Constitutive expression of catABC genes in the aniline-assimilating bacterium Rhodococcus species AN-22: production, purification, characterization and gene analysis of CatA, CatB and CatC  

PubMed Central

The aniline-assimilating bacterium Rhodococcus sp. AN-22 was found to constitutively synthesize CatB (cis,cis-muconate cycloisomerase) and CatC (muconolactone isomerase) in its cells growing on non-aromatic substrates, in addition to the previously reported CatA (catechol 1,2-dioxygenase). The bacterium maintained the specific activity of the three enzymes at an almost equal level during cultivation on succinate. CatB and CatC were purified to homogeneity and characterized. CatB was a monomer with a molecular mass of 44 kDa. The enzyme was activated by Mn2+, Co2+ and Mg2+. Native CatC was a homo-octamer with a molecular mass of 100 kDa. The enzyme was stable between pH 7.0 and 10.5 and was resistant to heating up to 90 °C. Genes coding for CatA, CatB and CatC were cloned and named catA, catB and catC respectively. The catABC genes were transcribed as one operon. The deduced amino acid sequences of CatA, CatB and CatC showed high identities with those from other Gram-positive micro-organisms. A regulator gene such as catR encoding a regulatory protein was not observed around the cat gene cluster of Rhodococcus sp. AN-22, but a possible relic of catR was found in the upstream region of catA. Reverse transcriptase-PCR and primer extension analyses showed that the transcriptional start site of the cat gene cluster was located 891 bp upstream of the catA initiation codon in the AN-22 strain growing on both aniline and succinate. Based on these data, we concluded that the bacterium constitutively transcribed the catABC genes and translated its mRNA into CatA, CatB and CatC. PMID:16156722

Matsumura, Eitaro; Sakai, Masashi; Hayashi, Katsuaki; Murakami, Shuichiro; Takenaka, Shinji; Aoki, Kenji

2005-01-01

230

Foundations of Our Constitution. Reflecting on September 11.  

ERIC Educational Resources Information Center

This lesson plan provides background on six significant historical documents: (1) "Meeting at Runnymede--The Story of King John and the Magna Carta"; (2) "The Mayflower Compact"; (3) "The Declaration of Independence and Natural Rights"; (4) "The Federalist Papers"; (5) "Aristotle: In Search of the Best Constitution"; and (6) "The Pledge of…

Constitutional Rights Foundation, Los Angeles, CA.

231

Everyday Constitutional Assessments and Their Relevance to Formal Assessments  

ERIC Educational Resources Information Center

Background: In anthropology and related disciplines, the term "assessment" refers to the everyday activities of ordinary people as they figure out what to do next given what others have just done. The assessments, in turn, constitute what is happening, whether in encounters between policeman and person in the street, or classroom lesson,…

Varenne, Herve

2014-01-01

232

Palatability of British trees to insects: constitutive and induced defences  

Microsoft Academic Search

Undamaged foliage of sixteen species of broadleaved trees was assessed for background (constitutive) palatability using larvae of Spodoptera littoralis (Lepidoptera) in laboratory bioassays. Palatability (dry weight consumed in 48 h) varied significantly between species and exhibited a four-fold range. Leaves of fifteen species were damaged artificially in the field and offered with control (undamaged) leaves to Spodoptera after 48 h

P. J. Edwards; S. D. Wratten; S. Greenwood

1986-01-01

233

Genomic expression during human myelopoiesis  

PubMed Central

Background Human myelopoiesis is an exciting biological model for cellular differentiation since it represents a plastic process where multipotent stem cells gradually limit their differentiation potential, generating different precursor cells which finally evolve into distinct terminally differentiated cells. This study aimed at investigating the genomic expression during myeloid differentiation through a computational approach that integrates gene expression profiles with functional information and genome organization. Results Gene expression data from 24 experiments for 8 different cell types of the human myelopoietic lineage were used to generate an integrated myelopoiesis dataset of 9,425 genes, each reliably associated to a unique genomic position and chromosomal coordinate. Lists of genes constitutively expressed or silent during myelopoiesis and of genes differentially expressed in commitment phase of myelopoiesis were first identified using a classical data analysis procedure. Then, the genomic distribution of myelopoiesis genes was investigated integrating transcriptional and functional characteristics of genes. This approach allowed identifying specific chromosomal regions significantly highly or weakly expressed, and clusters of differentially expressed genes and of transcripts related to specific functional modules. Conclusion The analysis of genomic expression during human myelopoiesis using an integrative computational approach allowed discovering important relationships between genomic position, biological function and expression patterns and highlighting chromatin domains, including genes with coordinated expression and lineage-specific functions. PMID:17683550

Ferrari, Francesco; Bortoluzzi, Stefania; Coppe, Alessandro; Basso, Dario; Bicciato, Silvio; Zini, Roberta; Gemelli, Claudia; Danieli, Gian Antonio; Ferrari, Sergio

2007-01-01

234

Mutations in the Arabidopsis Phosphoinositide Phosphatase Gene SAC9 Lead to Overaccumulation of PtdIns(4,5)P2 and Constitutive Expression of the Stress-Response Pathway1  

PubMed Central

Phosphoinositides (PIs) are signaling molecules that regulate cellular events including vesicle targeting and interactions between membrane and cytoskeleton. Phosphatidylinositol (PtdIns)(4,5)P2 is one of the best characterized PIs; studies in which PtdIns(4,5)P2 localization or concentration is altered lead to defects in the actin cytoskeleton and exocytosis. PtdIns(4,5)P2 and its derivative Ins(1,4,5)P3 accumulate in salt, cold, and osmotically stressed plants. PtdIns(4,5)P2 signaling is terminated through the action of inositol polyphosphate phosphatases and PI phosphatases including supressor of actin mutation (SAC) domain phosphatases. In some cases, these phosphatases also act on Ins(1,4,5)P3. We have characterized the Arabidopsis (Arabidopsis thaliana) sac9 mutants. The SAC9 protein is different from other SAC domain proteins in several ways including the presence of a WW protein interaction domain within the SAC domain. The rice (Oryza sativa) and Arabidopsis SAC9 protein sequences are similar, but no apparent homologs are found in nonplant genomes. High-performance liquid chromatography studies show that unstressed sac9 mutants accumulate elevated levels of PtdIns(4,5)P2 and Ins(1,4,5)P3 as compared to wild-type plants. The sac9 mutants have characteristics of a constitutive stress response, including dwarfism, closed stomata, and anthocyanin accumulation, and they overexpress stress-induced genes and overaccumulate reactive-oxygen species. These results suggest that the SAC9 phosphatase is involved in modulating phosphoinsitide signals during the stress response. PMID:15923324

Williams, Mary Elizabeth; Torabinejad, Javad; Cohick, Evan; Parker, Katherine; Drake, Elizabeth J.; Thompson, James E.; Hortter, Michelle; DeWald, Daryll B.

2005-01-01

235

Constitutive modeling for isotropic materials  

NASA Technical Reports Server (NTRS)

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.

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

1988-01-01

236

Growth of hormone-dependent MCF7 breast cancer cells is promoted by constitutive caveolin-1 whose expression is lost in an EGF-R-mediated manner during development of tamoxifen resistance  

Microsoft Academic Search

Caveolin-1 displays both tumour-suppressor and tumour-promoter properties in breast cancer. Using characterised preclinical\\u000a cell models for the transition of oestrogen-sensitive (WT-MCF-7 cells) to a tamoxifen-resistant (TAM-R cells) phenotype we\\u000a examined the role caveolin-1 in the development of hormone-resistant breast cancer. The WT-MCF-7 cells showed abundant expression\\u000a of caveolin-1 which potentiated oestrogen-receptor (ER?) signalling and promoted cell growth despite caveolin-1 mediating

Nicholas B. P. Thomas; Iain R. Hutcheson; Lee Campbell; Julia Gee; Kathryn M. Taylor; Robert I. Nicholson; Mark Gumbleton

2010-01-01

237

What is a Twentieth-Century Constitution?  

Microsoft Academic Search

At present, almost all of the constitutions in the world are twentieth-century constitutions; indeed, most of them were not adopted until the second half of the twentieth century. Accordingly, the eighteenth-century Constitution of the United States -- which includes the original constitution of 1787-89; the first ten amendments, adopted in 1791; and the Eleventh Amendment, adopted in 1798 -- antedates

Peter E. Quint

2007-01-01

238

Constitutional Exchange, Ideology and Democracy in America  

Microsoft Academic Search

Constitutional democracy in the United States emerged very gradually through a long series of constitutional bargains. A theory of constitutional exchange grounded in rational choice models provides a good explanation for the distinctive features of American constitutional history, as it does for much of the West, although it does less well at explaining the timing of some of changes. No

Roger D. Congleton

239

[Dynamics of local expression of connexin-43 and basic fibroblast growth factor receptors in patients with skin and soft-tissue infections against the background of diabetes mellitus type II].  

PubMed

Clinical results of wound healing dynamics were studied in 60 patients with soft-tissue infection against the background of diabetes mellitus type II. At the same time the study considered indices of intercellular contacts protein tissue expression such as connexin 43 (Cx43) and basic fibroblast growth factor receptors (bFGFR). The basic therapy of biopsy material of wound borders was applied. The reduction of bFGFR expression and the minor growth of Cx43 expression were observed. The pain syndrome proceeded for a long time and there were signs of perifocal inflammation, retard wound healing with granulation tissue. The application of combined method of ozone therapy which included autohemotherapy with ozone and an external management of wound by ozone-oxygen mixture facilitated to considerable shortening of inflammatory phase and regeneration. It was associated with increased Cx43 expression (in 1.9 times) in comparison with initial level and bFGFR was enlarged in 1.7 times to eighth day of postoperative period. PMID:25552106

2014-01-01

240

Importance of genetic background for risk of relapse shown in altered prefrontal cortex gene expression during abstinence following chronic alcohol intoxication  

PubMed Central

Alcoholism is a relapsing disorder associated with excessive consumption after periods of abstinence. Neuroadaptations in brain structure, plasticity and gene expression occur with chronic intoxication but are poorly characterized. Here we report identification of pathways altered during abstinence in prefrontal cortex, a brain region associated with cognitive dysfunction and damage in alcoholics. To determine the influence of genetic differences, an animal model was employed with widely divergent responses to alcohol withdrawal, the Withdrawal Seizure-Resistant (WSR) and Withdrawal Seizure-Prone (WSP) lines. Mice were chronically exposed to highly intoxicating concentrations of ethanol and withdrawn, then left abstinent for 21 days. Transcriptional profiling by microarray analyses identified a total of 562 genes as significantly altered during abstinence. Hierarchical cluster analysis revealed that the transcriptional response correlated with genotype/withdrawal phenotype rather than sex. Gene Ontology category overrepresentation analysis identified thyroid hormone metabolism, glutathione metabolism, axon guidance and DNA damage response as targeted classes of genes in low response WSR mice, with acetylation and histone deacetylase complex as highly dimorphic between WSR and WSP mice. Confirmation studies in WSR mice revealed both increased neurotoxicity by histopathologic examination and elevated T3 levels. Most importantly, relapse drinking was reduced by inhibition of thyroid hormone synthesis in dependent WSR mice compared to controls. These findings provide in vivo physiological and behavioral validation of the pathways identified. Combined, these results indicate a fundamentally distinct neuroadaptive response during abstinence in mice genetically selected for divergent withdrawal severity. Identification of pathways altered in abstinence may aid development of novel therapeutics for targeted treatment of relapse in abstinent alcoholics. PMID:21081154

Hashimoto, Joel G.; Forquer, Melissa R.; Tanchuck, Michelle A.; Finn, Deborah A.; Wiren, Kristine M.

2010-01-01

241

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  

SciTech Connect

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.

Al-Salman, Fadheela; Plant, Nick, E-mail: N.Plant@Surrey.ac.uk

2012-08-15

242

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).  

PubMed

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. PMID:21207033

Choi, Jun Young; Seo, Young Sam; Kim, Su Jin; Kim, Woo Taek; Shin, Jeong Sheop

2011-05-01

243

Cloning and functional characterization of an Arabidopsis nitrate transporter gene that encodes a constitutive component of low-affinity uptake.  

PubMed Central

The Arabidopsis CHL1 (AtNRT1) gene encodes an inducible component of low-affinity nitrate uptake, which necessitates a "two-component" model to account for the constitutive low-affinity uptake observed in physiological studies. Here, we report the cloning and characterization of a CHL1 homolog, AtNRT1:2 (originally named NTL1), with data to indicate that this gene encodes a constitutive component of low-affinity nitrate uptake. Transgenic plants expressing antisense AtNRT1:2 exhibited reduced nitrate-induced membrane depolarization and nitrate uptake activities in assays with 10 mM nitrate. Furthermore, transgenic plants expressing antisense AtNRT1:2 in the chl1-5 background exhibited an enhanced resistance to chlorate (7 mM as opposed to 2 mM for the chl1-5 mutant). Kinetic analysis of AtNRT1:2-injected Xenopus oocytes yielded a K(m) for nitrate of approximately 5.9 mM. In contrast to CHL1, AtNRT1:2 was constitutively expressed before and after nitrate exposure (it was repressed transiently only when the level of CHL1 mRNA started to increase significantly), and its mRNA was found primarily in root hairs and the epidermis in both young (root tips) and mature regions of roots. We conclude that low-affinity systems of nitrate uptake, like high-affinity systems, are composed of inducible and constitutive components and that with their distinct functions, they are part of an elaborate nitrate uptake network in Arabidopsis. PMID:10449574

Huang, N C; Liu, K H; Lo, H J; Tsay, Y F

1999-01-01

244

A high-throughput transient gene expression system for switchgrass (Panicum virgatum L.) seedlings  

Microsoft Academic Search

BACKGROUND: Grasses are relatively recalcitrant to genetic transformation in comparison to certain dicotyledons, yet they constitute some of the most important biofuel crops. Genetic transformation of switchgrass (Panicum virgatum L.) has previously been reported after cocultivation of explants with Agrobacterium and biolistics of embryogenic calli. Experiments to increase transient gene expression in planta may lead to stable transformation methods with

Xinlu Chen; Raymie Equi; Holly Baxter; Kyle Berk; Jin Han; Sujata Agarwal; Janice Zale

2010-01-01

245

3 CFR 8714 - Proclamation 8714 of September 16, 2011. Constitution Day and Citizenship Day, Constitution Week...  

Code of Federal Regulations, 2012 CFR

...Constitution Day and Citizenship Day, Constitution Week, 2011By the President of the United States of America A Proclamation In the...compromise, the delegates signed the Constitution of the United States. For more than two...

2012-01-01

246

3 CFR 8862 - Proclamation 8862 of September 13, 2012. Constitution Day and Citizenship Day, Constitution Week...  

Code of Federal Regulations, 2013 CFR

...Citizenship Day, Constitution Week, 2012By the President of the United States of America ...each year as “Constitution Week.” NOW...OBAMA, President of the United States of America, do...17, 2012, as Constitution Day and...

2013-01-01

247

3 CFR 8418 - Proclamation 8418 of September 16, 2009. Constitution Day and Citizenship Day, Constitution Week...  

Code of Federal Regulations, 2010 CFR

...Constitution Day and Citizenship Day, Constitution Week, 2009By the President of the United States of America A Proclamation The United States Constitution has withstood the test of time for more than two...

2010-01-01

248

3 CFR 9019 - Proclamation 9019 of September 16, 2013. Constitution Day and Citizenship Day, Constitution Week...  

...Citizenship Day, Constitution Week, 2013By the President of the United States of America ...each year as “Constitution Week.” NOW...OBAMA, President of the United States of America, do...17, 2013, as Constitution Day and...

2014-01-01

249

75 FR 57835 - Constitution Day and Citizenship Day, Constitution Week, 2010  

Federal Register 2010, 2011, 2012, 2013

...Day and Citizenship Day, Constitution Week, 2010 Proclamation 8563--National POW...Day and Citizenship Day, Constitution Week, 2010 By the President of the United States...Citizenship Day, and during Constitution Week, we commemorate the legacy passed...

2010-09-22

250

Written Constitution or None: Which Works Better?  

ERIC Educational Resources Information Center

Explores the differences between the U.S. Constitution and British constitutional law. Specifically examines the concept of the U.S. Bill of Rights in relation to the United Kingdom common law doctrine of parliamentary sovereignty. (BSR)

Cowen, Zelman

1987-01-01

251

Rethinking what constitutes suspended sediment  

NASA Astrophysics Data System (ADS)

Although cohesive suspended sediment is now known to be transported primarily as flocculated material, there is still a misconception of what constitutes suspended sediment. Flocs represent a complex matrix of microbial communities, organic particles (e.g. detritus, extracellular polymers and cellular debris), inorganic particles (e.g. clays and silts) and substantial interfloc spaces (pores), which allow for the retention or flow through of water. Flocculation results in significant alteration of the hydrodynamics of the constituent particles (by modifying their effective size, shape, density and porosity), thereby affecting the transport of sediment and associated contaminants. The composition and structure of a floc is in a continuous state of change as the medium in which it is transported provides the floc with further building materials, energy, nutrients and chemicals for biological growth, chemical reactions and morphological development. As such, a floc's physical (e.g. transport), chemical (e.g. contaminant adsorption) and biological (community development and contaminant biotransformation) behaviour are also in a continuous state of change, with concomitant effects on their aquatic environment as a whole. Although it is recognized that floc form will influence floc behaviour, there is still a basic lack of knowledge of the complex links between the structural components of a floc and how their individual properties and behaviours in combination with others will influence a floc's physical, chemical and biological behaviour. This paper provides a comprehensive conceptual model that links the many interrelated structural components of typical flocs and their interrelated behavioural aspects, in order to enhance our understanding of what constitutes suspended sediment.

Droppo, Ian G.

2001-06-01

252

Natural Law Reasoning and American Constitutional Discourse  

Microsoft Academic Search

This essay will suggest a link between the relative decline of natural law as a characteristic feature of American public debate, and concern over the possible decline of constitutional discourse. The first part of the essay will describe a particular understanding of constitutional discourse, sharing vital elements with natural law reasoning. Essential to this understanding is the notion that constitutional

Stephen A. Simon

2003-01-01

253

Antithetical Ethics: Kenneth Burke and the Constitution.  

ERIC Educational Resources Information Center

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…

Anderson, Virginia

1995-01-01

254

Florida Atlantic University Student Government Constitution  

E-print Network

the principles of the Constitution and laws of the United States and of the State of Florida, do ordainFlorida Atlantic University Student Government Constitution Preamble We the students of Florida and establish this Constitution for the Florida Atlantic University Student Government. Article I. Establishment

Fernandez, Eduardo

255

University of Florida Student Body Constitution  

E-print Network

rights under the constitutions and laws of the United States and State of Florida. Student government.--All political power granted by authorities authorized under the constitution, laws, and regulations of the StateUniversity of Florida Student Body Constitution Submitted by: David M. Kerner, Chairman 2009

Roy, Subrata

256

The Constitutional Politics of Charitable Choice.  

ERIC Educational Resources Information Center

Clarifies the constitutional issue raised by government efforts to cooperate with faith-based organizations, noting that while programs can be constitutional without necessarily being good public policy, too frequently political and constitutional controversies mix these two considerations, thus confusing those trying to understand and resolve…

Knippenberg, Joseph M.

2003-01-01

257

32 CFR 536.42 - Constitutional torts.  

Code of Federal Regulations, 2010 CFR

...2010-07-01 2010-07-01 true Constitutional torts. 536.42 Section 536...Processing of Claims § 536.42 Constitutional torts. A claim for violation...not cognizable under any subpart. A constitutional claim will be scrutinized in...

2010-07-01

258

American Focus on World Constitutions. Teacher's Guide.  

ERIC Educational Resources Information Center

This curriculum project was designed to familiarize high school students with their own constitutional roots while gaining a better understanding of governmental systems developed by other nations. The project uses the U.S. Constitution as a baseline for analyzing the constitutions of other nations, and is intended to supplement courses in such…

Holmes, Stanley T., III

259

Teaching about Democratic Constitutionalism. ERIC Digest.  

ERIC Educational Resources Information Center

There are more than 100 democracies in the world today. All but three of them--Great Britain, Israel, and New Zealand --have written constitutions. This digest examines the importance of constitutions and constitutionalism and the teaching of these concepts through the use of comparisons. The primary objectives of civic education for democratic…

Patrick, John J.

260

How Does the Constitution Secure Rights?  

ERIC Educational Resources Information Center

Third in a series designed to help prepare the nation for a thoughtful observance of the Constitutional bicentennial, this publication presents six essays addressing the issue of human rights and the Constitution. "How the Constitution Protects Our Rights: A Look at the Seminal Years" (Robert A. Rutland) focuses on James Madison's role in…

Goldwin, Robert A., Ed.; Schambra, William A., Ed.

261

Constitutively released adenosine diphosphate regulates proplatelet formation by human megakaryocytes  

PubMed Central

Background The interaction of adenosine diphosphate with its P2Y1 and P2Y12 receptors on platelets is important for platelet function. However, nothing is known about adenosine diphosphate and its function in human megakaryocytes. Design and Methods We studied the role of adenosine diphosphate and P2Y receptors on proplatelet formation by human megakaryocytes in culture. Results Megakaryocytes expressed all the known eight subtypes of P2Y receptors, and constitutively released adenosine diphosphate. Proplatelet formation was inhibited by the adenosine diphosphate scavengers apyrase and CP/CPK by 60-70% and by the P2Y12 inhibitors cangrelor and 2-MeSAMP by 50-60%, but was not inhibited by the P2Y1 inhibitor MRS 2179. However, the active metabolites of the anti-P2Y12 drugs, clopidogrel and prasugrel, did not inhibit proplatelet formation. Since cangrelor and 2-MeSAMP also interact with P2Y13, we hypothesized that P2Y13, rather than P2Y12 is involved in adenosine diphosphate-regulated proplatelet formation. The specific P2Y13 inhibitor MRS 2211 inhibited proplatelet formation in a concentration-dependent manner. Megakaryocytes from a patient with severe congenital P2Y12 deficiency showed normal proplatelet formation, which was inhibited by apyrase, cangrelor or MRS 2211 by 50-60%. The platelet count of patients with congenital delta-storage pool deficiency, who lack secretable adenosine diphosphate, was significantly lower than that of patients with other platelet function disorders, confirming the important role of secretable adenosine diphosphate in platelet formation. Conclusions This is the first demonstration that adenosine diphosphate released by megakaryocytes regulates their function by interacting with P2Y13. The clinical relevance of this not previously described physiological role of adenosine diphosphate and P2Y13 requires further exploration. PMID:22689668

Balduini, Alessandra; Di Buduo, Christian Andrea; Malara, Alessandro; Lecchi, Anna; Rebuzzini, Paola; Currao, Manuela; Pallotta, Isabella; Jakubowski, Joseph A.; Cattaneo, Marco

2012-01-01

262

Constitutive Theories for Thermoelastic Solids in Lagrangian Description Using Gibbs Potential  

E-print Network

of the constitution of the matter, the second law of thermodynamics, i.e. entropy inequality, must form the basis for all constitutive theories of the deforming matter to ensure thermodynamic equilibrium during the evolution [1, 2]. The entropy inequality expressed...

Mendoza, Yusshy

2012-08-31

263

Ending the Apartheid of the Closet: Sexual Orientation in the South African Constitutional Process  

Microsoft Academic Search

This paper will briefly examine the process of creating a new South African Constitution in the 1990s. Nearly a decade of talks preceded the adoption of South Africa's first multi-racial democratic constitution in 1994. These talks and the subsequent drafting conventions created an astonishing document, stunning in its novelty in South African history, in its expressed values, and in its

Eric C. Christiansen

2000-01-01

264

Constitutive relations associated with the Mott-Smith distribution function  

USGS Publications Warehouse

It is shown that the distribution function assumed by Mott-Smith determines a unique relation between heat flux, stress, and fluid velocity given by q = (3/2)??u, i.e., it provides a constitutive relation for heat flux, and it also determines a simple expression for this ratio of third-order central moments Q = . These expressions allow the equation of transfer for c x2 to be cast in a form that yields a nonlinear constitutive relation for stress. The results obtained from the Mott-Smith ansatz are compared with the theory of Baganoff and Nathenson and results from a numerical solution of the Boltzmann equation for shock-wave structure obtained by Hicks and Yen.

Nathenson, M.; Baganoff, D.

1973-01-01

265

Recruitment of Histone Deacetylase 3 to the Interferon-A Gene Promoters Attenuates Interferon Expression  

Microsoft Academic Search

BackgroundInduction of Type I Interferon (IFN) genes constitutes an essential step leading to innate immune responses during virus infection. Sendai virus (SeV) infection of B lymphoid Namalwa cells transiently induces the transcriptional expression of multiple IFN-A genes. Although transcriptional activation of IFN-A genes has been extensively studied, the mechanism responsible for the attenuation of their expression remains to be determined.Principal

Pierre Génin; Rongtuan Lin; John Hiscott; Ahmet Civas

2012-01-01

266

The E92K Melanocortin 1 Receptor Mutant Induces cAMP Production and Arrestin Recruitment but Not ERK Activity Indicating Biased Constitutive Signaling  

PubMed Central

Background The melanocortin 1 receptor (MC1R) constitutes a key regulator of melanism. Consequently, many naturally-occurring MC1R mutations are associated with a change in color. An example is the Glu-to-Lys substitution found at position II:20/2.60 in the top of transmembrane helix II which has been identified in melanic mice and several other species. This mutation induces a pronounced increase in MC1R constitutive activity suggesting a link between constitutive activity and melanism which is corroborated by the attenuation of ?-melanocyte stimulating hormone (?MSH) induced activation. However, the mechanism by which the mutation induces constitutive activity is currently not known. Methodology/Principal Findings Here we characterize the constitutive activity, cell surface expression and internalization of the mouse mutant, Mc1r E92K. As previously reported, only positively charged residues at position II:20/2.60 induced an increase in constitutive activity as measured by cAMP accumulation and CREB activation. Furthermore, the mutation induced a constitutive recruitment of ?-arrestin. This phenomenon is only observed in MC1R, however, as the equivalent mutations in MC2-5R had no effect on receptor signaling. Interestingly, the mutation did not induce constitutive ERK1/2 phosphorylation or increase the internalization rate indicating the constitutive activity to be biased. Finally, to identify regions of importance for the increased constitutive activity of Mc1r E92K, we employed a chimeric approach and identified G102 and L110 in the extracellular loop 1 to be selectively important for the constitutive activity as this, but not ?MSH-mediated activation, was abolished upon Ala substitution. Conclusions/Significance It is concluded that the E92K mutation induces an active conformation distinct from that induced by ?MSH and that the extracellular loop 1 is involved in maintaining this conformational state. In turn, the results suggest that in MC1R, which lacks an extracellular loop 2, the first extracellular loop may play a more prominent role during receptor activation than in general. PMID:21931793

Benned-Jensen, Tau; Mokrosinski, Jacek; Rosenkilde, Mette M.

2011-01-01

267

Finite gradient elasticity and plasticity: a constitutive mechanical framework  

NASA Astrophysics Data System (ADS)

Following a suggestion by Forest and Sievert (Acta Mech 160:71-111, 2003), a constitutive frame for a general gradient elastoplasticity for finite deformations is established. The basic assumptions are the principle of Euclidean invariance and the isomorphy of the elastic ranges. Both the elastic and the plastic laws include the first and the second deformation gradient. The starting point is an objective expression for the stress power.

Bertram, Albrecht

2014-11-01

268

The constitution of cometary nuclei  

NASA Technical Reports Server (NTRS)

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.

Whipple, F. L.

1977-01-01

269

Constitutive Activity of the Androgen Receptor  

PubMed Central

Prostate cancer (PCa) is the most frequently diagnosed cancer in the United States. The androgen receptor (AR) signaling axis is central to all stages of PCa pathophysiology and serves as the main target for endocrine-based therapy. The most advanced stage of the disease, castration resistant prostate cancer (CRPC), is presently incurable and accounts for most PCa mortality. In this review, we highlight the mechanisms by which the AR signaling axis can bypass endocrine-targeted therapies and drive progression of CRPC. These mechanisms include alterations in growth factor, cytokine, and inflammatory signaling pathways, altered expression or activity of transcriptional co-regulators, AR point mutations, and AR gene amplification leading to AR protein overexpression. Additionally, we will discuss the mechanisms underlying the synthesis of constitutively active AR splice variants (AR-Vs) lacking the COOH-terminal ligand binding domain, as well as the role and regulation of AR-Vs in supporting therapeutic resistance in CRPC. Finally, we summarize the ongoing development of inhibitors targeting discrete AR functional domains as well as the status of new biomarkers for monitoring the AR signaling axis in patients. PMID:24931201

Chan, Siu Chiu; Dehm, Scott M.

2014-01-01

270

The venom composition of the parasitic wasp Chelonus inanitus resolved by combined expressed sequence tags analysis and proteomic approach  

Microsoft Academic Search

BACKGROUND: Parasitic wasps constitute one of the largest group of venomous animals. Although some physiological effects of their venoms are well documented, relatively little is known at the molecular level on the protein composition of these secretions. To identify the majority of the venom proteins of the endoparasitoid wasp Chelonus inanitus (Hymenoptera: Braconidae), we have randomly sequenced 2111 expressed sequence

Bruno Vincent; Martha Kaeslin; Thomas Roth; Manfred Heller; Julie Poulain; François Cousserans; Johann Schaller; Marylčne Poirié; Beatrice Lanzrein; Jean-Michel Drezen; Sébastien JM Moreau

2010-01-01

271

FindLaw Constitutional Law Center  

NSDL National Science Digital Library

This offering from FindLaw (last discussed in the January 15, 1999 Scout Report) focuses on the US Constitution. The site is composed of two primary sections. The first offers an annotated Constitution, commentary on selected topics with links to key sections of the Constitution or The Federalist Papers, a history of the Constitution, and brief biographies of notable "Founding Fathers" and delegates to the Constitutional Convention. The second section highlights the Third Branch of government, with a history of the Supreme Court, landmark decisions regarding civil rights with the full opinions, a summary and analysis of the criminal law and procedure decisions of the October 1998 Supreme Court Term, and several links for current Supreme Court news and information. Both sections also feature a search engine. Additional resources at the site include a Town Hall section that explores Hot Topics (currently flag burning) and message boards on a wide variety of Constitutional issues.

1994-01-01

272

FindLaw Constitutional Law Center  

NSDL National Science Digital Library

This latest offering from FindLaw (last discussed in the January 15, 1999 Scout Report) focuses on the US Constitution. The site is composed of two primary sections. The first offers an annotated Constitution, commentary on selected topics with links to key sections of the Constitution or The Federalist Papers, a history of the Constitution, and brief biographies of notable "Founding Fathers" and delegates to the Constitutional Convention. The second section highlights the Third Branch of government, with a history of the Supreme Court, landmark decisions regarding civil rights with the full opinions, a summary and analysis of the criminal law and procedure decisions of the October 1998 Supreme Court Term, and several links for current Supreme Court news and information. Both sections also feature a search engine. Additional resources at the site include a Town Hall section that explores Hot Topics (currently flag burning) and message boards on a wide variety of Constitutional issues.

273

Testing of constitutive models in LAME.  

SciTech Connect

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 analysis, providing problems that highlight the behavior of various constitutive models to the engineer can be of great benefit. LAME is currently implemented in the Sierra based solid mechanics codes Adagio [3] and Presto [4]. The constitutive models in LAME are available in both codes. Due to the nature of a transient dynamics code--e.g. Presto--it is difficult to test a constitutive model due to inertia effects that show up in the solution. Therefore the testing of constitutive models is primarily done in Adagio. All of the test problems detailed in this report are run in Adagio. It is the goal of the constitutive model test suite to provide a useful service for the constitutive model developer, application code developer and engineer that uses the application code. Due to the conflicting needs and tight time constraints on solid mechanics code development, no requirements exist for implementing test problems for constitutive models. Model developers are strongly encouraged to provide test problems and document those problems, but given the choice of having a model without a test problem or no model at all, certain requirements must be kept loose. A flexible code development environment, especially with regards to research and development in constitutive modeling, is essential to the success of such an environment. This report provides documentation of a number of tests for the constitutive models in LAME. Each section documents a separate test with a brief description of the model, the test problem and the results. This report is meant to be updated periodically as more test problems are created and put into the test suite.

Hammerand, Daniel Carl; Scherzinger, William Mark

2007-09-01

274

An integrative expression vector for Actinosynnema pretiosum  

PubMed Central

Background The Actinomycete Actinosynnema pretiosum ssp. auranticum has commercial importance due to its production of ansamitocin P-3 (AP-3), a potent antitumor agent. One way to increase AP-3 production would be to constitutively express selected genes so as to relieve bottlenecks in the biosynthetic pathway; however, an integrative expression vector for A. pretiosum is lacking. The aim of this study was to construct a vector for heterologous gene expression in A. pretiosum. Results A series of integrative expression vectors have been made with the following features: the IS117 transposase from Streptomyces coelicolor, the constitutive ermE* promoter from Saccharopolyspora erythraea, different ribosome-binding site (RBS) sequences and xylE as a translational reporter. Positive E. coli clones and A. pretiosum transconjugants were assayed by catechol. pAP42, containing an E. coli consensus RBS, and pAP43, containing an asm19 RBS, gave strong and moderate gene expression, respectively. In addition, an operon construct capable of multi-gene expression was created. Plasmid integration sites in transconjugants were investigated and four different sites were observed. Although the most common integration site was within a putative ORF with sequence similarity to NADH-flavin reductase, AP-3 levels and cell growth of transconjugants were unaffected. Conclusion A set of integrative vectors for constitutive gene expression in A. pretiosum has been constructed. Gene translation is easily determined by colorimetric assay on an agar plate. The vectors are suitable for studies relating to AP-3 biosynthesis as they do not affect AP-3 production. PMID:17956638

Goh, Shan; Camattari, Andrea; Ng, Daniel; Song, Ruth; Madden, Kevin; Westpheling, Janet; Wong, Victor VT

2007-01-01

275

Ratification of European Union Constitution Hits a Roadblock  

NSDL National Science Digital Library

This past week, the ratification of the European Union's constitution met with some resistance as voters in both the Netherlands and France both voted "no" to approving the document. Some analysts have suggested that this rejection of the constitution is indicative of the fact that there is an increasing unease about the process of unification more generally in various countries across the continent. Pundits and analysts also agreed that there was no one defining reason why voters rejected the constitution. In France, those voting "no" seemed to range from those on the far right to the far left, and interestingly enough, included a large proportion of young people. It is worth noting that a number of opponents (particularly those on the political left) of ratification continue to make the claim that the constitution promotes a type of unencumbered capitalism that is akin to the type of socioeconomic mode of governance practiced in the United Kingdom and the United States. It will be sometime before the full impact of this recent development is understood, but it is certainly a story that is worthy of a closer look. The first link will take users to coverage of the ongoing EU Constitution process offered by the Voice of America from this past Monday. The second link will take visitors to a fine news piece from the Guardian that provides some insight into the recent rejection of the EU Constitution by Dutch voters. The third link leads to an article from Deutsche Welle that discusses the implications of these latest developments in Europe. The fourth link leads to a very thorough set of analyses of the ratification process as reported by National Public Radio. From this main page, visitors can also learn about the history of the EU Constitution and read some essential background material about the document. The fifth link, offered by the BBC, provides a quick introduction to the Constitution, along with providing a link to the full text of the document. The final link leads to the homepage of the Historical Archives of the European Union, where visitors can review primary documents related to the EU's development and also learn about utilizing their various collections.

276

The Constitution in the Twentieth Century.  

ERIC Educational Resources Information Center

Investigates the development of the United States Constitution in the twentieth century up to and including the Burger Court. Contends that interpreting the Constitution is an important issue of our times. Consequently argues that we should teach students about the development of this document. (RKM)

Murphy, Paul L.

1987-01-01

277

An Internet Guide to Teaching the Constitution  

ERIC Educational Resources Information Center

In 2004, Congress passed and President Bush signed legislation designating every September 17 as Constitution Day and suggesting that all schools teach about the Constitution on that day. There are innumerable websites, countless lesson plans and a multitude of other teaching resources on the topic. Teachers were inundated with brochures,…

Risinger, C. Frederick

2005-01-01

278

Guy Verhofstadt The new European Constitution  

E-print Network

Guy Verhofstadt The new European Constitution from Laeken to Rome Humboldt-Reden zu Europa 25 The new European Constitution ­ from Laeken to Rome 8 �ber den Autor 20 #12;#12;5 Richard Schröder belgische Revolution von 1830, durch die Belgien endgültig unabhängig wurde, von der feinsten #12;6 Art

Röder, Beate

279

The Constitution at 200: Celebration Amidst Controversy.  

ERIC Educational Resources Information Center

Current debates about the Constitution fall into the three following categories: (1) reappraisals of consitutional origins; (2) disagreements on hermeneutical principles used in contemporary applications; and (3) discussions of contemporary events whose consequences for law and political stability could not have been foreseen by the Constitution's…

Collins, Sheila D.

1987-01-01

280

The Intelligence Function And the Constitution  

Microsoft Academic Search

Strategic intelligence is requisite to national security. But fitting an intelligence function, with its ancillary covert action and counterintelligence activities, into the U.S. constitutional framework is an enduring and perhaps unsolvable problem. Conduct of the government's clandestine operations overseas is an executive function requiring extraordinary secrecy. Yet the Constitution requires that the president share with Congress decisions about intelligence policy,

Harry Howe Ransom

1987-01-01

281

Contragate, Covert Action, and the Constitution.  

ERIC Educational Resources Information Center

The "Iran-Contragate" hearings were indicative of serious constitutional crisis. Analyzes and disputes following three defenses used by President Reagan to justify his actions in affair: he did not know or approve; it was not a violation of law; and his authority over foreign affairs is given in war powers clause of the Constitution--Congress…

Ratner, Michael

1987-01-01

282

Naked Land Transfers and American Constitutional Development  

Microsoft Academic Search

The constitutional prohibition on naked land transfers, laws granting to B property that belonged to A, played a far greater role in American constitutional development than is generally realized. The Marshall and Taney Courts heard numerous cases in which government officials were accused of expropriating private property, typically by legislative oversight rather than by deliberate intent. When resolving these cases,

Mark A. Graber

2000-01-01

283

Gender, Nation and the Common Law Constitution  

Microsoft Academic Search

This article argues that the common law constitution can be thought of as the working out of a tradition within which notions of gender, national identity and citizenship are conveyed and secured. It looks at the making and interpretation of Commonwealth Caribbean Constitutions in the latter half of the twentieth century. It shows how the language of the common law

Tracy Robinson

2008-01-01

284

Constitutive modeling of inelastic anisotropic material response  

NASA Technical Reports Server (NTRS)

A constitutive equation was developed to predict the inelastic thermomechanical response of single crystal turbine blades. These equations are essential for developing accurate finite element models of hot section components and contribute significantly to the understanding and prediction of crack initiation and propagation. The method used was limited to unified state variable constitutive equations. Two approaches to developing an anisotropic constitutive equation were reviewed. One approach was to apply the Stouffer-Bodner representation for deformation induced anisotropy to materials with an initial anisotropy such as single crystals. The second approach was to determine the global inelastic strain rate from the contribution of the slip in each of the possible crystallographic slip systems. A three dimensional finite element is being developed with a variable constitutive equation link that can be used for constitutive equation development and to predict the response of an experiment using the actual specimen geometry and loading conditions.

Stouffer, D. C.

1984-01-01

285

Documents from the Continental Congress and the Constitutional Convention  

NSDL National Science Digital Library

The Library of Congress American Memory Project has been producing high-quality digital collections for a decade, and one of their first projects has recently been updated and is worthy of a second (or first) look. This particular collection brings together a number of documents, including dozens of broadsides related to the work of the Continental Congress, dating from 1774 to 1788. The majority of these items contain extracts of the journals of Congress, resolutions, proclamations, treaties, and other congressional proceedings. Other documents in the collection include early printed versions of the United States Constitution and the Declaration of Independence. Those persons who are seeking a bit more background information will want to take a look at two timelines that cover related events from the 1764 Sugar Act to the official ratification of the Constitution in 1788. Finally, the site also contains a nice essay titled â??To Form a More Perfect Union.â?ť.

286

3 CFR 8562 - Proclamation 8562 of September 16, 2010. Constitution Day and Citizenship Day, Constitution Week...  

Code of Federal Regulations, 2011 CFR

...Convention in Philadelphia established a Constitution for the United States of America, signing the finished charter...solemn oath to “support and defend the Constitution and laws of the United States of America.” In so doing, they...

2011-01-01

287

76 FR 58705 - Constitution Day and Citizenship Day, Constitution Week, 2011  

Federal Register 2010, 2011, 2012, 2013

...Constitution Day and Citizenship Day, Constitution Week, 2011 Proclamation 8715--National Employer Support of the Guard and Reserve Week, 2011 Proclamation 8716--National Farm Safety and Health Week, 2011 Proclamation 8717--National...

2011-09-21

288

Closing in on the constitution of consciousness  

PubMed Central

The science of consciousness is a nascent and thriving field of research that is founded on identifying the minimally sufficient neural correlates of consciousness. However, I have argued that it is the neural constitution of consciousness that science seeks to understand and that there are no evident strategies for distinguishing the correlates and constitution of (phenomenal) consciousness. Here I review this correlation/constitution distinction problem and challenge the existing foundations of consciousness science. I present the main analyses from a longer paper in press on this issue, focusing on recording, inhibition, stimulation, and combined inhibition/stimulation strategies, including proposal of the Jenga analogy to illustrate why identifying the minimally sufficient neural correlates of consciousness should not be considered the ultimate target of consciousness science. Thereafter I suggest that while combined inhibition and stimulation strategies might identify some constitutive neural activities—indeed minimally sufficient constitutive neural activities—such strategies fail to identify the whole neural constitution of consciousness and thus the correlation/constitution distinction problem is not fully solved. Various clarifications, potential objections and related scientific and philosophical issues are also discussed and I conclude by proposing new foundational claims for consciousness science. PMID:25452738

Miller, Steven M.

2014-01-01

289

A biosynthetic regulated secretory pathway in constitutive secretory cells  

PubMed Central

It has frequently been proposed that while the constitutive secretory pathway is present in all cells, the regulated secretory pathway is found only in specialized cells such as neuronal, endocrine, or exocrine types. In this study we provide evidence that suggests that this distinction is not as restrictive as proposed. We have identified a population of post-Golgi storage vesicles in several constitutive secretory cells using [35S]SO4-labeled glycosaminoglycan (GAG) chains as a marker. A fraction of this pool of vesicles can undergo exocytosis in response to stimuli such as cytoplasmic Ca2+ and phorbol esters. The effect of Ca2+ was demonstrated both in intact cells in the presence of the ionophore A23187 and in streptolysin-O-permeabilized semi-intact cells. N-ethylmaleiimide, under conditions known to block regulated and constitutive secretion, inhibited the stimulated secretion from these cells, suggesting that the observed release of labeled GAG chains was not due to a leakage artefact. Subcellular fractionation revealed that the stored GAG chains were in low-density membrane granules (d approximately 1.12 g/ml), whose size was greater than that of synaptic- like vesicles found in PC12 cells. In addition, in CHO cells that express epitope-tagged rab 3D, the labeled GAG chains were found to cofractionate with the exogenous rab protein. When expressed in the regulated cell line AtT-20, this tagged rab protein was found to colocalize with ACTH-containing dense-core granules by indirect immunofluorescence. Taken together, these results provide evidence for the presence of a cryptic regulated secretory pathway in "constitutive" cells and suggest that the regulated secretory pathway is more widespread amongst different cell types than previously believed. PMID:8682857

1996-01-01

290

Constitutively active UVR8 photoreceptor variant in Arabidopsis  

PubMed Central

Arabidopsis thaliana UV RESISTANCE LOCUS 8 (UVR8) is a UV-B photoreceptor that initiates photomorphogenic responses underlying acclimation and UV-B tolerance in plants. UVR8 is a homodimer in its ground state, and UV-B exposure results in its instantaneous monomerization followed by interaction with CONSTITUTIVELY PHOTOMORPHOGENIC 1 (COP1), a major factor in UV-B signaling. UV-B photoreception by UVR8 is based on intrinsic tryptophan aromatic amino acid residues, with tryptophan-285 as the main chromophore. We generated transgenic plants expressing UVR8 with a single amino acid change of tryptophan-285 to alanine. UVR8W285A appears monomeric and shows UV-B–independent interaction with COP1. Phenotypically, the plants expressing UVR8W285A exhibit constitutive photomorphogenesis associated with constitutive activation of target genes, elevated levels of anthocyanins, and enhanced, acclimation-independent UV-B tolerance. Moreover, we have identified COP1, REPRESSOR OF UV-B PHOTOMORPHOGENESIS 1 and 2 (RUP1 and RUP2), and the SUPPRESSOR OF PHYA-105 (SPA) family as proteins copurifying with UVR8W285A. Whereas COP1, RUP1, and RUP2 are known to directly interact with UVR8, we show that SPA1 interacts with UVR8 indirectly through COP1. We conclude that UVR8W285A is a constitutively active UVR8 photoreceptor variant in Arabidopsis, as is consistent with the crucial importance of monomer formation and COP1 binding for UVR8 activity. PMID:24277841

Heijde, Marc; Binkert, Melanie; Yin, Ruohe; Ares-Orpel, Florence; Rizzini, Luca; Van De Slijke, Eveline; Persiau, Geert; Nolf, Jonah; Gevaert, Kris; De Jaeger, Geert; Ulm, Roman

2013-01-01

291

Ordered Rate Constitutive Theories in Eulerian Description  

E-print Network

law, Carreau-Yasuda model for viscosity, power law, Sutherland law etc. for temperature dependent material coefficients are a subset of the general ordered rate constitutive theories of order n for thermofluids (iii) Maxwell model, Oldroyd-B model...

Nunez, Daniel

2012-12-31

292

Constitutional Growth Delay and Learning Problems.  

ERIC Educational Resources Information Center

Records of children admitted to a growth clinic and follow-up questionnaires were examined to determine the incidence of learning problems in children with constitutional growth delay and those with familial short stature. (Author/PHR)

Gold, Ruth F.

1978-01-01

293

The constitutive modeling of nitrile elastomers  

E-print Network

THE CONSTITUTIVE MODELING OF NITRILE ELASTOMERS A Thesis by LUMAS TERENCE HELAIRE Submitted to the OIIice of Graduate Studies of Texas A&M University in partial fulfdlment of the requirements for the degree of MASTER OF SCIENCE December 1996... Major Subject: Mechanical Engineering THE CONSTITUTIVE 1VIODELING OF NITRILE ELASTOMERS A Thesis by LUMAS TERENCE HELAIRE Submitted to the OIRce of Graduate Studies of Texas A&M University in partial fulfillment of the requirements for the degree...

Helaire, Lumas Terence

1996-01-01

294

The American Political and Constitutional Context  

Microsoft Academic Search

\\u000a Textually, the Constitution on positive rights is “voiceless as fear in a wide wilderness” (Keats, 281). Indeed, the original\\u000a Constitution—what Gladstone famously called “The most wonderful work ever struck off at a given time by the brain and purpose\\u000a of man” (Gladstone, “Kin”, 185)—made no mention of rights at all. Rights appearednot among the six justifications in the Preamble\\u000a nor

Thomas Halper

295

Answers to the Urbino Questions support material WHAT CONSTITUTES THE OZONE (*) BACKGROUND OVER EUROPE?  

E-print Network

continental European man-made emissions of gases leading to the production of ozone (O3) were switched off ozone at the Earth's surface, for 1997, simulated by the GEOS-CHEM model. The top panel shows ozone from to the non-linear behaviour of ozone production related to the complexity of the atmospheric chemical

296

Radio brightness distribution within cosmic background fluctuations  

NASA Astrophysics Data System (ADS)

The angular fluctuations delta T in the microwave background temperature constitute a random process. Calculations are performed for the number of 'pips' above a base level that might be produced by primordial scalar density perturbations or gravitational waves. The flux density expected for typical pips is determined, as well as the mean solid angle subtended by a pip that would exceed a level three times the dispersion in delta T.

Sazhin, M. V.

1985-08-01

297

Background Subtraction Techniques  

Microsoft Academic Search

Background subtraction is a commonly used class of techniques for segmenting out objects of interest in a scene for applications such as surveillance. This paper surveys a repre- sentative sample of the published techiques for background subtraction, and analyses them with respect to three important attributes: foreground detection; background maintenance; and postprocessing.

Alan M. McIvor

298

Constitutional Rights of Children. Prepared for the Subcommittee on the Constitution of the Committee on the Judiciary, United States Senate, by the American Law Division, Congressional Research Service of the Library of Congress.  

ERIC Educational Resources Information Center

This paper reviews the rights of children as defined by the Constitution of the United States and summarizes a series of Supreme Court decisions which have defined the protections afforded to children by the Constitution. A short historical overview of the legal status of children is provided as background for the report. It is suggested that the…

Library of Congress, Washington, DC. Congressional Research Service.

299

Rate constitutive theories for ordered thermofluids  

NASA Astrophysics Data System (ADS)

The paper considers developments of constitutive theories in Eulerian description for compressible as well as incompressible ordered homogeneous and isotropic thermofluids in which the deviatoric Cauchy stress tensor and the heat vector are functions of density, temperature, temperature gradient, and the convected time derivatives of the strain tensors of up to a desired order. The fluids described by these constitutive theories are called ordered thermofluids due to the fact that the constitutive theories for the deviatoric Cauchy stress tensor and heat vector are dependent on the convected time derivatives of the strain tensor up to a desired order, the highest order of the convected time derivative of the strain tensor in the argument tensors defines the `order of the fluid'. The admissibility requirement necessitates that the constitutive theories for the stress tensor and heat vector satisfy conservation laws, hence, in addition to conservation of mass, balance of momenta, and conservation of energy, the second law of thermodynamics, that is, Clausius-Duhem inequality must also be satisfied by the constitutive theories or be used in their derivations. If we decompose the total Cauchy stress tensor into equilibrium and deviatoric components, then Clausius-Duhem inequality and Helmholtz free energy density can be used to determine the equilibrium stress in terms of thermodynamic pressure for compressible fluids and in terms of mechanical pressure for incompressible fluids, but the second law of thermodynamics provides no mechanism for deriving the constitutive theories for the deviatoric Cauchy stress tensor. In the development of the constitutive theories in Eulerian description, the covariant and contravariant convected coordinate systems, and Jaumann measures are natural choices. Furthermore, the mathematical models for fluids require Eulerian description in which material point displacements are not measurable. This precludes the use of displacement gradients, that is, strain measures, in the development of the constitutive theories. It is shown that compatible conjugate pairs of convected time derivatives of the deviatoric Cauchy stress and strain measures in co-, contravariant, and Jaumann bases in conjunction with the theory of generators and invariants provide a general mathematical framework for the development of constitutive theories for ordered thermofluids in Eulerian description. This framework has a foundation based on the basic principles and axioms of continuum mechanics but the resulting constitutive theories for the deviatoric Cauchy stress tensor must satisfy the condition of positive work expanded, a requirement resulting from the entropy inequality. The paper presents a general theory of constitutive equations for ordered thermofluids which is then specialized, assuming first-order thermofluids, to obtain the commonly used constitutive theories for compressible and incompressible generalized Newtonian and Newtonian fluids. It is demonstrated that the constitutive theories for ordered thermofluids of all orders are indeed rate constitutive theories. We have intentionally used the term `thermofluids' as opposed to `thermoviscous fluids' due to the fact that the constitutive theories presented here describe a broader group of fluids than Newtonian and generalized Newtonian fluids that are commonly referred as thermoviscous fluids.

Surana, K. S.; Nunez, D.; Reddy, J. N.; Romkes, A.

2013-09-01

300

The Cosmic Background Explorer.  

ERIC Educational Resources Information Center

Outlines the Cosmic Background Explorer (COBE) mission to measure celestial radiation. Describes the instruments used and experiments involving differential microwave radiometers, and a far infrared absolute spectrophotometer. (YP)

Gulkis, Samuel; And Others

1990-01-01

301

Effect of Reduced Myristoylated Alanine-Rich C Kinase Substrate Expression on Hippocampal Mossy Fiber Development and Spatial Learning in Mutant Mice: Transgenic Rescue and Interactions with Gene Background  

Microsoft Academic Search

The myristoylated alanine-rich C kinase substrate (MARCKS) is a prominent protein kinase C (PKC) substrate in brain that is expressed highly in hippocampal granule cells and their axons, the mossy fibers. Here, we examined hippocampal infrapyramidal mossy fiber (IP-MF) limb length and spatial learning in heterozygous Macs mutant mice that exhibit an ≈ 50% reduction in MARCKS expression relative to

R. K. McNamara; D. J. Stumpo; L. M. Morel; M. H. Lewis; E. K. Wakeland; P. J. Blackshear; R. H. Lenox

1998-01-01

302

Acquired mutations in phage lambda genes O or P that enable constitutive expression of a cryptic lambdaN+cI[Ts]cro- prophage in E. coli cells shifted from 30 degreesC to 42 degreesC, accompanied by loss of immlambda and Rex+ phenotypes and emergence of a non-immune exclusion-state.  

PubMed

The majority of bacteria, which carry the N+-cI857[Ts]-cro--O+-P+ fragment of lambda genome, are killed when derepressed by shifting from 30 degreesC to 42 degreesC. Among rare survivors, we observed a proportion of colony-forming units (cfu) that retained the typical immlambda-immunity phenotype when grown at 30 degreesC; however, when shifted from 30 degreesC to 42 degreesC, they lost lambda immunity and acquired a non-immune exclusion-state (Nie phenotype). We also found that the immlambda survivor cfu quickly lost their Rex+ exclusion phenotype (as measured by T4rII plating inhibition) when shifted from 30 degreesC to 42 degreesC, even though they produced CII, which stimulates pE-cI-rexA-rexB transcription. The Nie phenotype was characterized by an inhibition of plating of the homoimmune phage, lambdawt, and the heteroimmune phage, lambdaimm434. However, lambdavir and spontaneous mutants of lambdawt (lambdase mutations localized within oR) escaped the Nie exclusion-state and plated efficiently on lawns of Nie cfu at 42 degreesC. Thus, we examined the scope of the Nie exclusion-state toward lambda mutants blocked for lysogeny, and lambda hybrids substituted for immunity or replication genes. Phage like lambdawt, competent for lysogeny, were severely excluded compared to some mutants of lambda defective for lysogeny. Among this latter type, there was high variance in the Nie exclusion of various cI mutants; some of which were not excluded. The Nie exclusion-state was attributed to the constitutive expression of the defective lambda fragment in the survivor cfu, made possible by the acquired replication defect(s). We characterized, both genetically and physically, the mutations in the defective integrated lambda prophage that permitted growth of the survivor cfu at 42 degreesC. In five of seven survivor cfu, we identified IS2 insertions within lambda genes O and P that can block replication initiation from the lambda fragment. The remaining survivor cfu had multiple base substitutions within the C-terminal end of O and N-terminal half of P, the majority of which were silent. In some of these mutants, either an ochre nonsense mutation or a single-base frameshift deletion inactivated P. PMID:9858705

Hayes, S; Hayes, C; Bull, H J; Pelcher, L A; Slavcev, R A

1998-11-26

303

Constitutive Androstane Receptor Ligands Modulate the Anti-Tumor Efficacy of Paclitaxel in Non-Small Cell Lung Cancer Cells  

PubMed Central

Background Lung tumors are the leading cause of cancer deaths worldwide and paclitaxel has proven to be useful for patients with lung cancer, however, acquired resistance is a major problem. To overcome this problem, one promising option is the use of Constitutive Androstane Receptor (CAR) ligands in combination with chemotherapeutics against cancer cells. Therefore, we wish to elucidate the effects of CAR ligands on the antineoplastic efficacy of paclitaxel in lung cancer cells. Methodology/Principal Findings Our results from cell viability assays exposing CAR agonist or inverse-agonist to mouse and human lung cancer cells modulated the antineoplastic effect of paclitaxel. The CAR agonists increased the effect of Paclitaxel in 6 of 7 lung cancer cell lines, whereas the inverse-agonist had no effect on paclitaxel cytotoxicity. Interestingly, the mCAR agonist TCPOBOP enhanced the expression of two tumor suppressor genes, namely WT1 and MGMT, which were additively enhanced in cells treated with CAR agonist in combination with paclitaxel. Also, in silico analysis showed that both paclitaxel and CAR agonist TCPOBOP docked into the mCAR structure but not the inverse agonist androstenol. Paclitaxel per se increases the expression of CAR in cancer cells. At last, we analyzed the expression of CAR in two public independent studies from The Cancer Genome Atlas (TCGA) of Non Small Cell Lung Cancer (NSCLC). CAR is expressed in variable levels in NSCLC samples and no association with overall survival was noted. Conclusions/Significance Taken together, our results demonstrated that CAR agonists modulate the antineoplastic efficacy of paclitaxel in mouse and human cancer cell lines. This effect was probably related by the enhanced expression of two tumor suppressor genes, viz. WT1 and MGMT. Most of NSCLC cases present CAR gene expression turning it possible to speculate the use of CAR modulation by ligands along with Paclitaxel in NSCLC therapy. PMID:24959746

Fukumasu, Heidge; Rochetti, Arina L.; Pires, Pedro R. L.; Silva, Edson R.; Mesquita, Ligia G.; Strefezzi, Ricardo F.; De Carvalho, Daniel D.; Dagli, Maria L.

2014-01-01

304

Background music reactive games  

Microsoft Academic Search

In this paper, we discuss the concept of games that react to their background music. Instead of limiting the player to a fixed set of songs, the background music can be any song chosen from the player's own music collection. Due to the relative simplicity of such existing game titles, we wanted to explore the potential of the concept more

Juha Arrasvuori; Jukka Holm

2010-01-01

305

Correlators in nontrivial backgrounds  

SciTech Connect

Operators in N=4 super Yang-Mills theory with an R-charge of O(N{sup 2}) are dual to backgrounds which are asymtotically AdS{sub 5}xS{sup 5}. In this article we develop efficient techniques that allow the computation of correlation functions in these backgrounds. We find that (i) contractions between fields in the string words and fields in the operator creating the background are the field theory accounting of the new geometry, (ii) correlation functions of probes in these backgrounds are given by the free field theory contractions but with rescaled propagators and (iii) in these backgrounds there are no open string excitations with their special end point interactions; we have only closed string excitations.

Mello Koch, Robert de [National Institute for Theoretical Physics, Department of Physics and Centre for Theoretical Physics, University of the Witwatersrand, Wits, 2050 (South Africa); Stellenbosch Institute for Advanced Studies, Stellenbosch (South Africa); Ives, Norman; Stephanou, Michael [National Institute for Theoretical Physics, Department of Physics and Centre for Theoretical Physics, University of the Witwatersrand, Wits, 2050 (South Africa)

2009-01-15

306

77 FR 57981 - Constitution Day and Citizenship Day, Constitution Week, 2012  

Federal Register 2010, 2011, 2012, 2013

...Day and Citizenship Day, Constitution Week, 2012 Presidential Documents Federal Register...Day and Citizenship Day, Constitution Week, 2012 By the President of the United States...contributions of all their members. This week, we reflect on the basic rights and...

2012-09-18

307

Dynamic constitutional frameworks for DNA biomimetic recognition.  

PubMed

Linear and cross-linked dynamic constitutional frameworks generated from reversibly interacting linear PEG/core constituents and cationic sites shed light on the dominant coiling versus linear DNA binding behaviours, closer to the histone DNA binding wrapping mechanism. PMID:25500992

Catana, Romina; Barboiu, Mihail; Moleavin, Ioana; Clima, Lilia; Rotaru, Alexandru; Ursu, Elena-Laura; Pinteala, Mariana

2015-01-22

308

A constitutive model for partially saturated soils  

Microsoft Academic Search

The Paper presents a constitutive model for describing the stress-strain behaviour of partially saturated soils. The model is formulated within the framework of hardening plasticity using two iode- pendent sets of stress variables: the excess of total stress over air pressure and the suction. The mode1 is able to represent, in a consistent and unified manner, many of the fundamental

E. E. ALONSO; A. GENS; A. JOSA

1990-01-01

309

Social Work Students' Society Constitution November 2005  

E-print Network

Social Work Students' Society Constitution ­ November 2005 Preamble The Social Work Students operating out of the University of Regina's Faculty of Social Work. Mission The SWSS exists as a venue for social work students to gather and pursue common goals and interests. The SWSS shall have as an over

Argerami, Martin

310

What Constitutes Good Mathematics Teaching: Teachers' Perspectives.  

ERIC Educational Resources Information Center

Throughout the history of mathematics education, many studies have attempted to determine what constitutes "good" mathematics teaching and how such teaching can be put into practice. Missing in the literature, however, are the voices of secondary mathematics teachers and how they define good mathematics teaching. This study contributes to the…

Wilson, Patricia S.; Cooney, Thomas J.; Stinson, David; Drumm, Chris

311

The Constitutive Equation for Membrane Tether Extraction  

PubMed Central

Membrane tethers or nanotubes play a critical role in a variety of cellular and subcellular processes such as leukocyte rolling and intercellular mass transport. The current constitutive equations that describe the relationship between the pulling force and the tether velocity during tether extraction have serious limitations. Here we propose a new phenomenological constitutive equation that captures all known characteristics of nanotube formation, including nonlinearity, nonzero threshold force, and possible negative tether velocity. We used tether extraction from endothelial cells as a prototype to illustrate how to obtain the material constants in the constitutive equation. With the micropipette aspiration technique, we measured tether pulling forces at both positive and negative tether velocities. We also determined the threshold force of 55 pN experimentally for the first time. This new constitutive equation unites two established ones and provides us a unified platform to better understand not only the physiological role of tether extraction during leukocyte rolling and intercellular or intracellular transport, but also the physics of membrane tether growth or retraction. PMID:20614242

Chen, Yong; Yao, Da-Kang; Shao, Jin-Yu

2010-01-01

312

Constitutive laws, tensorial invariance and chocolate cake  

NASA Astrophysics Data System (ADS)

Although constitutive modeling is a well-established branch of mathematics which has found wide industrial application, geophysicists often do not take full advantage of its known results. We present a synopsis of the theory of constitutive modeling, couched in terms of the ‘simple material’, which has been extensively studied and is complex enough to include most of the correct models proposed to describe the behavior of geological materials. Critical in the development of the theory are various invariance requirements, the principal ones being coordinate invariance, peer group invariance (isotropy), and frame-indifference. Each places distinet restrictions on constitutive equations. A noncomprehensive list of properly invariant and commonly used constitutive equations is given. To exemplify use of the equations, we consider two problems in detail: steady extension, which models the commonly performed constant strain rate triaxial test, and simple shearing. We note that each test is so restricted kinematically that only the most trivial aspects of material behavior are manifested in these tests, no matter how complex the material. Furthermore, the results of one test do not generally determine the results of the other.

Rundle, John B.; Passman, S. L.

1982-04-01

313

Constitutional Rights: A View from Mt. Healthy.  

ERIC Educational Resources Information Center

In their 1977 decision, "Mt. Healthy City School District Board of Education v. Doyle," the Supreme Court held that if a government employee who had a reasonable expectation of continued employment is dismissed and can show that his dismissal was motivated by his taking actions that are constitutionally protected, the employing agency must prove…

Gee, E. Gordon

314

Application of symbolic computations to the constitutive modeling of structural materials  

NASA Technical Reports Server (NTRS)

In applications involving elevated temperatures, the derivation of mathematical expressions (constitutive equations) describing the material behavior can be quite time consuming, involved and error-prone. Therefore intelligent application of symbolic systems to faciliate this tedious process can be of significant benefit. Presented here is a problem oriented, self contained symbolic expert system, named SDICE, which is capable of efficiently deriving potential based constitutive models in analytical form. This package, running under DOE MACSYMA, has the following features: (1) potential differentiation (chain rule), (2) tensor computations (utilizing index notation) including both algebraic and calculus; (3) efficient solution of sparse systems of equations; (4) automatic expression substitution and simplification; (5) back substitution of invariant and tensorial relations; (6) the ability to form the Jacobian and Hessian matrix; and (7) a relational data base. Limited aspects of invariant theory were also incorporated into SDICE due to the utilization of potentials as a starting point and the desire for these potentials to be frame invariant (objective). The uniqueness of SDICE resides in its ability to manipulate expressions in a general yet pre-defined order and simplify expressions so as to limit expression growth. Results are displayed, when applicable, utilizing index notation. SDICE was designed to aid and complement the human constitutive model developer. A number of examples are utilized to illustrate the various features contained within SDICE. It is expected that this symbolic package can and will provide a significant incentive to the development of new constitutive theories.

Arnold, Steven M.; Tan, H. Q.; Dong, X.

1990-01-01

315

The cosmic neutrino background  

NASA Technical Reports Server (NTRS)

The cosmic neutrino background is expected to consist of relic neutrinos from the big bang, of neutrinos produced during nuclear burning in stars, of neutrinos released by gravitational stellar collapse, and of neutrinos produced by cosmic ray interactions with matter and radiation in the interstellar and intergalactic medium. Formation of baryonic dark matter in the early universe, matter-antimatter annihilation in a baryonic symmetric universe, and dark matter annihilation could have also contributed significantly to the cosmic neutrino background. The purpose of this paper is to review the properties of these cosmic neutrino backgrounds, the indirect evidence for their existence, and the prospects for their detection.

Dar, Arnon

1991-01-01

316

The GLAST Background Model  

NASA Astrophysics Data System (ADS)

In order to estimate the ability of the GLAST/LAT to reject unwanted background of charged particles, optimize the on-board processing, size the required telemetry and optimize the GLAST orbit, we developed a detailed model of the background particles that would affect the LAT. In addition to the well-known components of the cosmic radiation, we included splash and reentrant components of protons, electrons (e+ and e-) from 10 MeV and beyond as well as the albedo gamma rays produced by cosmic ray interactions with the atmosphere. We made estimates of the irreducible background components produced by positrons and hadrons interacting in the multilayered micrometeorite shield and spacecraft surrounding the LAT and note that because the orbital debris has increased, the shielding required and hence the background are larger than were present in EGRET. Improvements to the model are currently being made to include the east-west effect.

Ormes, J. F.; Atwood, W.; Burnett, T.; Grove, E.; Longo, F.; McEnery, J.; Mizuno, T.; Ritz, S.

2007-07-01

317

The Cosmic Background Radiation  

E-print Network

We summarise the current status of cosmic microwave background spectrum and anisotropy measurements, and their theoretical interpretation. This is the update of the mini-review for the 1997 web-version of the Review of Particle Properties.

George Smoot; Douglas Scott

1997-11-08

318

IR-background database  

Microsoft Academic Search

The Swedish Defence Research Agency (FOI) has recently performed systematic measurements in order to establish an IR-background database. It will be used for a wide range of applications and provide a basis for the modeling of IR-background properties of Swedish terrain. Experimental data like this is also necessary for the validation of methods and programs for synthetic IR-scene simulation. The

Claes Nelsson; Paer Nilsson; Roland Lindell; Emma Bernhardsson

2001-01-01

319

An elastoplastic damage constitutive model for concrete  

NASA Astrophysics Data System (ADS)

An elastoplastic damage constitutive model to simulate nonlinear behavior of concrete is presented. Similar to traditional plastic theory, the irreversible deformation is modeled in effective stress space. In order to better describe different stiffness degradation mechanisms of concrete under tensile and compressive loading conditions, two damage variables, i.e., tension and compression are introduced, to quantitatively evaluate the degree of deterioration of concrete structure. The rate dependent behavior is taken into account, and this model is derived firmly in the framework of irreversible thermodynamics. Fully implicit backward-Euler algorithm is suggested to perform constitutive integration. Numerical results of the model accord well with the test results for specimens under uniaxial tension and compression, biaxial loading and triaxial loading. Failure processes of double-edge-notched (DEN) specimen are also simulated to further validate the proposed model.

Liu, Jun; Lin, Gao; Zhong, Hong

2013-04-01

320

A constitutive model for an overlay coating  

NASA Technical Reports Server (NTRS)

Coatings are frequently applied to gas turbine blades and vanes to provide protection against oxidation and corrosion. The results of an experimental and analytical study to develop a constitutive model for an overlay coating is presented. Specimens were machined from a hot isostatically pressed billet of PWA 286. The tests consisted of isothermal stress relaxation cycles with monotonically increasing maximum strain and were conducted at various temperatures. The results were used to calculate the constants for various constitutive models, including the classical, the Walker isotropic, a simplified Walker, and Stowell models. A computerized regression analysis was used to calculate model constants from the data. The best fit was obtained for the Walker model, with the simplified Walker and classical models close behind.

Nissley, D. M.; Swanson, G. A.

1988-01-01

321

Laboratory investigation of constitutive property scaling behavior  

SciTech Connect

Because many constitutive rock properties must be measured at one scale but applied at another, scaling behavior is an issue facing many applied disciplines, including the petroleum industry. A research program has been established to investigate and a quantify scaling behavior through systematic physical experimentation, with the aim of developing and testing models that describe scaling behavior in a quantitative manner. Scaling of constitutive rock properties is investigated through physical experimentation involving the collection of gas-permeability data measured over a range of discrete scales. The approach is to systematically isolate those factors that influence property scaling and investigate their relative contributions to overall scaling behavior. Two blocks of rock, each exhibiting differing heterogeneity structure. have recently been examined. The two samples were found to yield different scaling behavior, as exhibited by changes in the distribution functions and semi-variograms. Simple models have been fit to the measured scaling behavior that are of similar functional form but of different magnitude.

Tidwell, V.C.

1994-10-01

322

Constitutive Modeling of Crosslinked Nanotube Materials  

NASA Technical Reports Server (NTRS)

A non-linear, continuum-based constitutive model is developed for carbon nanotube materials in which bundles of aligned carbon nanotubes have varying amounts of crosslinks between the nanotubes. The model accounts for the non-linear elastic constitutive behavior of the material in terms of strain, and is developed using a thermodynamic energy approach. The model is used to examine the effect of the crosslinking on the overall mechanical properties of variations of the crosslinked carbon nanotube material with varying degrees of crosslinking. It is shown that the presence of the crosslinks has significant effects on the mechanical properties of the carbon nanotube materials. An increase in the transverse shear properties is observed when the nanotubes are crosslinked. However, this increase is accompanied by a decrease in axial mechanical properties of the nanotube material upon crosslinking.

Odegard, G. M.; Frankland, S. J. V.; Herzog, M. N.; Gates, T. S.; Fay, C. C.

2004-01-01

323

Constitutive modeling for single crystal superalloys  

NASA Technical Reports Server (NTRS)

A crystallographic approach to constitutive modeling of single crystal superalloys is discussed. The approach is based on identifying the active slip planes and slip directions. The shear stresses are computed on each of the slip planes from applied stress components. The slip rate is then computed on each slip system and the microscopic inelastic strain rates are the sum of the slip in the individual slip systems. The constitutive model was implemented in a finite element code using twenty noted isoparametric solid elements. Constants were determined for octahedral and cube slip systems. These constants were then used to predict tension-compression asymmetry and fatigue loops. Other data was used to model the tensile and creep response.

Stouffer, Donald C.; Dame, L. Thomas; Jayaraman, N.

1985-01-01

324

Rapid implementation of advanced constitutive models  

NASA Astrophysics Data System (ADS)

This paper presents a methodology based on the NICE integration scheme [1, 2] for simple and rapid numerical implementation of a class of plasticity constitutive models. In this regard, an algorithm is purposely developed for the implementation of newly developed advanced constitutive models into explicit finite element framework. The methodology follows the organization of the problem state variables into an extended form, which allows the constitutive models' equations to be organized in such a way, that the algorithm can be optionally extended with minimal effort to integrate also evolution equations related to a description of other specific phenomena, such as damage, distortional hardening, phase transitions, degradation etc. To confirm simplicity of the program implementation, computational robustness, effectiveness and improved accuracy of the implemented integration algorithm, a deep drawing simulation of the cylindrical cup is considered as the case study, performed in ABAQUS/Explicit. As a fairly complex considered model, the YLD2004-18p model [3, 4] is first implemented via external subroutine VUMAT. Further, to give additional proof of the simplicity of the proposed methodology, a combination of the YLD2004-18p model and Gurson-Tvergaard-Needleman model (GTN) is considered. As demonstrated, the implementation is really obtained in a very simple way.

Starman, Bojan; Halilovi?, Miroslav; Vrh, Marko; Štok, Boris

2013-12-01

325

A constitutive mechanical model for energetic materials  

SciTech Connect

Cookoff modeling of energetic materials has traditionally addressed reactive heat flow with the goal of defining the onset of runaway combustion behavior. Current modeling efforts are now aimed toward predicting the violence of the event. Combined thermal, chemical, and mechanical response must be modeled, since confinement results in pressure buildup which can breach confinement or enhance gas-phase combustion rates leading to runaway combustion behavior. Thermally induced stresses can also cause gaps which inhibit heat flow. These mechanical effects must also be included in cookoff modeling. A new reactive elastic-plastic constitutive model for micromechanical response has been developed which represents a stress-strain relation for reacting materials such as explosives, propellants, pyrotechnics, or burning foams. This micromechanical model is based on bubble mechanics. A local force balance, with mass continuity constraints, forms the basis of the constitutive model requiring input of temperature and reacted fraction. This constitutive material model has been incorporated into a quasistatic mechanics code, SANTOS. To provide temperature and reacted gas fraction, the thermal-chemical solver, XCHEM, has been coupled to SANTOS. This paper summarizes the development of the micromechanical model with material property estimates for conventional energetic materials. This study shows that large pressures can arise from small reacted fractions which implies that cookoff modeling must consider the strong interaction between thermochemistry and mechanics.

Hobbs, M.L.; Baer, M.R.; Gross, R.J.

1994-06-01

326

Constitutively active Rheb induces oncogenic transformation  

PubMed Central

Rheb (Ras homolog enriched in brain) is a component of the phosphatidylinositol 3-kinase (PI3K)–target of rapamycin (TOR) signaling pathway, functioning as a positive regulator of TOR. Constitutively active mutants of Rheb induce oncogenic transformation in cell culture. The transformed cells are larger and contain more protein than their normal counterparts. They show constitutive phosphorylation of the ribosomal protein S6 kinase (S6K) and the eukaryotic initiation factor 4E-binding protein 1 (4E-BP1), two downstream targets of TOR. The TOR-specific inhibitor rapamycin strongly interferes with transformation induced by constitutively active Rheb, suggesting that TOR activity is essential for the oncogenic effects of mutant Rheb. Rheb-induced transformation is also dependent on a C-terminal farnesylation signal that mediates localization to a cellular membrane. An engineered N-terminal myristylation signal can substitute for the farnesylation. Immunofluorescence localizes wild type (wt) and mutant Rheb to vesicular structures in the cytoplasm, overlapping with the endoplasmic reticulum (ER). PMID:18521078

Jiang, Hao; Vogt, Peter K.

2008-01-01

327

Constitutive modeling for isotropic materials (HOST)  

NASA Technical Reports Server (NTRS)

The results of the first year of work on a program to validate unified constitutive models for isotropic materials utilized in high temperature regions of gas turbine engines and to demonstrate their usefulness in computing stress-strain-time-temperature histories in complex three-dimensional structural components. The unified theories combine all inelastic strain-rate components in a single term avoiding, for example, treating plasticity and creep as separate response phenomena. An extensive review of existing unified theories is given and numerical methods for integrating these stiff time-temperature-dependent constitutive equations are discussed. Two particular models, those developed by Bodner and Partom and by Walker, were selected for more detailed development and evaluation against experimental tensile, creep and cyclic strain tests on specimens of a cast nickel base alloy, B19000+Hf. Initial results comparing computed and test results for tensile and cyclic straining for temperature from ambient to 982 C and strain rates from 10(exp-7) 10(exp-3) s(exp-1) are given. Some preliminary date correlations are presented also for highly non-proportional biaxial loading which demonstrate an increase in biaxial cyclic hardening rate over uniaxial or proportional loading conditions. Initial work has begun on the implementation of both constitutive models in the MARC finite element computer code.

Lindholm, Ulric S.; Chan, Kwai S.; Bodner, S. R.; Weber, R. M.; Walker, K. P.; Cassenti, B. N.

1984-01-01

328

Expression of endoglucanases in Pichia pastoris under control of the GAP promoter  

PubMed Central

Background Plant-derived biomass is a potential alternative to fossil feedstocks for a greener economy. Enzymatic saccharification of biomass has been studied extensively and endoglucanases have been found to be a prerequisite for quick initial liquefaction of biomass under industrial conditions. Pichia pastoris, widely used for heterologous protein expression, can be utilized for fungal endoglucanase production. The recently marketed PichiaPink™ expression system allows for rapid clone selection, and employs the methanol inducible AOX1 promoter to ensure high protein expression levels. However, methanol is toxic and poses a fire hazard, issues which become more significant at an industrial scale. It is possible to eliminate these risks and still maintain high productivity by switching to the constitutive GAP promoter. Results In the present study, a plasmid carrying the constitutive GAP promoter was created for PichiaPink™. We then studied expression of two endoglucanases, AfCel12A from Aspergillus fumigatus and TaCel5A from Thermoascus aurantiacus, regulated by either the AOX1 promoter or the GAP promoter. Initial experiments in tubes and small bioreactors showed that the levels of AfCel12A obtained with the constitutive promoter were similar or higher, compared to the AOX1 promoter, whereas the levels of TaCel5A were somewhat lower. After optimization of cultivation conditions using a 15-l bioreactor, the recombinant P. pastoris strains utilizing the GAP promoter produced ca. 3–5 g/l of total secreted protein, with CMCase activity equivalent to 1200 nkat/ml AfCel12A and 170 nkat/ml TaCel5A. Conclusions We present a strategy for constitutive recombinant protein expression in the novel PichiaPink™ system. Both AfCel12A and TaCel5A were successfully expressed constitutively in P. pastoris under the GAP promoter. Reasonable protein levels were reached after optimizing cultivation conditions. PMID:24742273

2014-01-01

329

The United States Constitution as an Incomplete Text  

Microsoft Academic Search

Viewing the Constitution of the United States of America as a political text leads to the application of techniques of textual analysis when reading it. Textual analysis shows the Constitution to be incomplete, both as a constitution and as a founding instrument of the federal union. A complete text of a constitution for the United States requires inclusion of the

Donald S. Lutz

1988-01-01

330

Power: Constitutional Update. Bar/School Partnership Programs Series.  

ERIC Educational Resources Information Center

The fourth in a special series of handbooks dealing with constitutional themes, this document looks at power in the context of the U.S. Constitution. "The Constitution's Prescription for Freedom" (L. Peach) examines the separation of powers provided for in the Constitution. "The Concept of Power" (C. Roach) is a series of strategies, some using…

American Bar Association, Chicago, IL. Special Committee on Youth Education for Citizenship.

331

Constitutional Thinness and Anorexia Nervosa: A Possible Misdiagnosis?  

PubMed Central

Clinical and biological aspects of restrictive anorexia nervosa (R-AN) are well documented. More than 10,000 articles since 1911 and more than 600 in 2013 have addressed R-AN psychiatric, somatic, and biological aspects. Genetic background, ineffectiveness of appetite regulating hormones on refeeding process, bone loss, and place of amenorrhea in the definition are widely discussed and reviewed. Oppositely, constitutional thinness (CT) is an almost unknown entity. Only 32 articles have been published on this topic since 1953. Similar symptoms associating low body mass index, low fat, and bone mass are reported in both CT and R-AN subjects. Conversely, menses are preserved in CT women and almost the entire hormonal profile is normal, except for leptin and PYY. The aim of the present review is to alert the clinician on the confusing clinical presentation of these two situations, a potential source of misdiagnosis, especially since R-AN definition has changed in DSM5. PMID:25368605

Estour, Bruno; Galusca, Bogdan; Germain, Natacha

2014-01-01

332

High constitutive activity of native H3 receptors regulates histamine neurons in brain  

Microsoft Academic Search

Some G-protein-coupled receptors display `constitutive activity', that is, spontaneous activity in the absence of agonist. This means that a proportion of the receptor population spontaneously undergoes an allosteric transition, leading to a conformation that can bind G proteins. The process has been shown to occur with recombinant receptors expressed at high density, and\\/or mutated, but also non-mutated recombinant receptors expressed

Séverine Morisset; Agnčs Rouleau; Xavier Ligneau; Florence Gbahou; Joël Tardivel-Lacombe; Holger Stark; Walter Schunack; C. Robin Ganellin; Jean-Michel Arrang

2000-01-01

333

78 FR 57777 - Constitution Day and Citizenship Day, Constitution Week, 2013  

Federal Register 2010, 2011, 2012, 2013

...nascent country. They met in a time of economic hardship and passionate debate, but with the understanding that while controversy...defines our history; with amendments that trace our national journey, the Constitution bears witness to how far we have...

2013-09-19

334

Involvement of the GTP binding protein Rho in constitutive endocytosis in Xenopus laevis oocytes  

Microsoft Academic Search

To study an endocytotic role of the GTP- binding protein RhoA in Xenopus oocytes, we have monitored changes in the surface expression of sodium pumps, the surface area of the oocyte and the uptake of the fluid-phase marker inulin. Xenopus oocytes possess intracellular sodium pumps that are continuously ex- changed for surface sodium pumps by constitutive endo- and exocytosis. Injection

Giinther Schmalzing; Hans-Peter Richter; Alexander Hansen; Wolfgang Schwarz; Ingo Just

1995-01-01

335

Rumex acetosa Y chromosomes: constitutive or facultative heterochromatin?  

PubMed

Condensed Y chromosomes in Rumex acetosa L. root-tip nuclei were studied using 5-azaC treatment and immunohistochemical detection of methylated histones. Although Y chromosomes were decondensed within root meristem in vivo, they became condensed and heteropycnotic in roots cultured in vitro. 5-azacytidine (5-azaC) treatment of cultured roots caused transitional dispersion of their Y chromosome bodies, but 7 days after removal of the drug from the culture medium, Y heterochromatin recondensed and again became visible. The response of Rumex sex chromatin to 5-azaC was compared with that of condensed segments of pericentromeric heterochromatin in Rhoeo spathacea (Sw.) Steam roots. It was shown that Rhoeo chromocentres, composed of AT-rich constitutive heterochromatin, did not undergo decondensation after 5-azaC treatment. The Y-bodies observed within male nuclei of R. acetosa were globally enriched with H3 histone, demethylated at lysine 4 and methylated at lysine 9. This is the first report of histone tail-modification in condensed sex chromatin in plants. Our results suggest that the interphase condensation of Y chromosomes in Rumex is facultative rather than constitutive. Furthermore, the observed response of Y-bodies to 5-azaC may result indirectly from demethylation and the subsequent altered expression of unknown genes controlling tissue-specific Y-inactivation as opposed to the global demethylation of Y-chromosome DNA. PMID:16201317

Mosio?ek, Magdalena; Pasierbek, Pawe?; Malarz, Janusz; Mo?, Maria; Joachimiak, Andrzej J

2005-01-01

336

Outside-In Signalling Generated by a Constitutively Activated Integrin ?IIb?3 Impairs Proplatelet Formation in Human Megakaryocytes  

PubMed Central

Background The interaction of megakaryocytes with matrix proteins of the osteoblastic and vascular niche is essential for megakaryocyte maturation and proplatelet formation. Fibrinogen is present in the vascular niche and the fibrinogen receptor ?IIb?3 is abundantly expressed on megakaryocytes, however the role of the interaction between fibrinogen and ?IIb?3 in proplatelet formation in humans is not yet fully understood. We have recently reported a novel congenital macrothrombocytopenia associated with a heterozygous mutation of the ?3 subunit of ?IIb?3. The origin of thrombocytopenia in this condition remains unclear and this may represent an interesting natural model to get further insight into the role of the megakaryocyte fibrinogen receptor in megakaryopoiesis. Methodology/Principal Findings Patients' peripheral blood CD45+ cells in culture were differentiated into primary megakaryocytes and their maturation, spreading on different extracellular matrix proteins, and proplatelet formation were analyzed. Megakaryocyte maturation was normal but proplatelet formation was severely impaired, with tips decreased in number and larger in size than those of controls. Moreover, megakaryocyte spreading on fibrinogen was abnormal, with 50% of spread cells showing disordered actin distribution and more evident focal adhesion points than stress fibres. Integrin ?IIb?3 expression was reduced but the receptor was constitutively activated and a sustained, and substrate-independent, activation of proteins of the outside-in signalling was observed. In addition, platelet maturation from preplatelets was impaired. Conclusions/Significance Our data show that constitutive activation of ?IIb?3-mediated outside-in signalling in human megakaryocytes negatively influences proplatelet formation, leading to macrothombocytopenia. PMID:22539947

Bury, Loredana; Malara, Alessandro; Gresele, Paolo; Balduini, Alessandra

2012-01-01

337

Cosmic Microwave Background  

NSDL National Science Digital Library

In this lesson, students explore the cosmic microwave background to understand why it permeates the universe and why it peaks as microwave radiation. Students should be able to explain that the origin of the background radiation is the uniform thermal radiation of the big bang and that the radiation produced was evenly distributed around the small early universe, causing it to permeate today's universe. This activity is part of the Cosmic Times teachers guide and is intended to be used in conjunction with the 1965 Cosmic Times Poster.

338

The Cosmic Background Explorer  

NASA Technical Reports Server (NTRS)

The Cosmic Background Explorer (CBE), NASA's cosmological satellite which will observe a radiative relic of the big bang, is discussed. The major questions connected to the big bang theory which may be clarified using the CBE are reviewed. The satellite instruments and experiments are described, including the Differential Microwave Radiometer, which measures the difference between microwave radiation emitted from two points on the sky, the Far-Infrared Absolute Spectrophotometer, which compares the spectrum of radiation from the sky at wavelengths from 100 microns to one cm with that from an internal blackbody, and the Diffuse Infrared Background Experiment, which searches for the radiation from the earliest generation of stars.

Gulkis, Samuel; Lubin, Philip M.; Meyer, Stephan S.; Silverberg, Robert F.

1990-01-01

339

The cosmic microwave background  

NASA Technical Reports Server (NTRS)

Recent limits on spectral distortions and angular anisotropies in the cosmic microwave background are reviewed. The various backgrounds are described, and the theoretical implications are assessed. Constraints on inflationary cosmology dominated by cold dark matter (CDM) and on open cosmological models dominated by baryonic dark matter (BDM), with, respectively, primordial random phase scale-invariant curvature fluctuations or non-gaussian isocurvature fluctuations are described. More exotic theories are addressed, and I conclude with the 'bottom line': what theorists expect experimentalists to be measuring within the next two to three years without having to abandon their most cherished theories.

Silk, Joseph

1991-01-01

340

Radioactive Decay 1. Background  

E-print Network

Radioactive Decay 1. Background It is well known that many nuclei are unstable and are transformed into other nuclear species by means of either alpha decay or beta decay. The rate at which those radioactive on the number N of radioactive nuclei in the sample and also on the probability for each nucleus to decay

Elster, Charlotte

341

Country background Forest history  

E-print Network

33 Country background Forest history During the Gallo-Roman period (1st­4th century AD), forests this proportion decreased dramatically to only 15­17 % of the land area. This residual forest was then severely Colbert's Forest Ordinance was instituted in 1669 a gradual restoration took place. High forests produced

Paris-Sud XI, Université de

342

David Smith Academic background  

E-print Network

David Smith Academic background Ph.D. in Mathematics (Algebra), Université de Sherbrooke, Canada project program (I. Assem, F. Bergeron, C. Reutenauer, D. Smith) $132,000 ($44,000 per year for 3 years. Schiffler and D. Smith, Friezes, strings and cluster variables, to appear in Glasgow Mathematcal Journal. 2

343

PANDEMIC INFLUENZA background briefing  

E-print Network

PANDEMIC INFLUENZA background briefing Biomedicine Forum 5 November 2008 compiled by David Evans, Dave Carr, David Lynn and Phil Green Transmission electron micrograph of Influenza A virus (Wellcome influenza!' Page 2 #12;Consequences of an influenza pandemic THE PANDEMIC THREAT DEATH If the next pandemic

Rambaut, Andrew

344

Local microwave background radiation  

E-print Network

An inquiry on a possible local origin for the Microwave Background Radiation is made. Thermal MBR photons are contained in a system called {\\it magnetic bottle} which is due to Earth magnetic field and solar wind particles, mostly electrons. Observational tests are anticipated.

Domingos Soares

2014-11-13

345

Gauging the cosmic microwave background  

E-print Network

We provide a new derivation of the anisotropies of the cosmic microwave background (CMB), and find an exact expression that can be readily expanded perturbatively. Close attention is paid to gauge issues, with the motivation to examine the effect of super-Hubble modes on the CMB. We calculate a transfer function that encodes the behaviour of the dipole, and examine its long-wavelength behaviour. We show that contributions to the dipole from adiabatic super-Hubble modes are strongly suppressed, even in the presence of a cosmological constant, contrary to claims in the literature. We also introduce a naturally defined CMB monopole, which exhibits closely analogous long-wavelength behaviour. We discuss the geometrical origin of this super-Hubble suppression, pointing out that it is a simple reflection of adiabaticity, and hence argue that it will occur regardless of the matter content.

J. P. Zibin; Douglas Scott

2008-08-14

346

Deleterious background selection with recombination  

SciTech Connect

An analytic expression for the expected nucleotide diversity is obtained for a neutral locus in a region with deleterious mutation and recombination. Our analytic results are used to predict levels of variation for the entire third chromosome of Drosophila melanogaster. The predictions are consistent with the low levels of variation that have been observed at loci near the centromeres of the third chromosome of D. melanogaster. However, the low levels of variation observed near the tips of this chromosome are not predicted using currently available estimates of the deleterious mutation rate and of selection coefficients. If considerably smaller selection coefficients are assumed, the low observed levels of variation at the tips of the third chromosome are consistent with the background selection model. 33 refs., 4 figs., 1 tab.

Hudson, R.R. [Univ. of California, Irvine, CA (United States); Kaplan, N.L. [National Institute of Environmental Health Sciences, Research Triangle Park, NC (United States)

1995-12-01

347

Constitutional and thermal defects in nickel aluminides  

SciTech Connect

The formation energies of intrinsic point defects and the interaction energies of possible defect pairs in NiAl are calculated from first principles within an order-N, locally self-consistent Green's function method in conjunction with the multipole electrostatic corrections to the atomic sphere approximation. The theory correctly reproduces the ground-state properties of the off-stoichiometric NiAl alloys. The constitutional defects (antisite Ni atoms in Ni-rich and Ni vacancies in Al-rich NiAl) are shown to form ordered structures in the ground state, in which the defects of the same kind tend to avoid each other at the shortest separation distance on their sublattice. A mean-field theory is applied to calculate the equilibrium concentrations of thermal defects. The statistics of thermal defects is interpreted in terms of dominant composition-conserving complex defects which are shown to be triple defects in Ni-rich and nearly stoichiometric NiAl. In the Al-rich region a novel thermal excitation dominates where two constitutional Ni vacancies are replaced by one antisite Al atom. The number of vacancies, as well as the total number of point defects decrease with temperature in Al-rich NiAl. The boundary between the two regions is treated analytically. The vacancy concentration exhibits a minimum in its temperature dependence at the boundary. Similar analysis is applied to study constitutional and thermal defects in Ni{sub 3}Al as a function of concentration is in excellent agreement with recent experimental data.

Korzhavyi, P.A.; Abrikosov, I.A.; Johansson, B.

1999-07-01

348

Numerically abnormal chromosome constitutions in humans  

SciTech Connect

Chapter 24, discusses numerically abnormal chromosome constitutions in humans. This involves abnormalities of human chromosome number, including polyploidy (when the number of sets of chromosomes increases) and aneuploidy (when the number of individual normal chromosomes changes). Chapter sections discuss the following chromosomal abnormalities: human triploids, imprinting and uniparental disomy, human tetraploids, hydatidiform moles, anomalies caused by chromosomal imbalance, 13 trisomy (D{sub 1} trisomy, Patau syndrome), 21 trisomy (Down syndrome), 18 trisomy syndrome (Edwards syndrome), other autosomal aneuploidy syndromes, and spontaneous abortions. The chapter concludes with remarks on the nonrandom participation of chromosomes in trisomy. 69 refs., 3 figs., 4 tabs.

NONE

1993-12-31

349

Constitutive modeling for isotropic materials (HOST)  

NASA Technical Reports Server (NTRS)

The results of the third year of work on a program which is part of the NASA Hot Section Technology program (HOST) are presented. The goals of this program are: (1) the development of unified constitutive models for rate dependent isotropic materials; and (2) the demonstration of the use of unified models in structural analyses of hot section components of gas turbine engines. The unified models selected for development and evaluation are those of Bodner-Partom and of Walker. A test procedure was developed for assisting the generation of a data base for the Bodner-Partom model using a relatively small number of specimens. This test procedure involved performing a tensile test at a temperature of interest that involves a succession of strain-rate changes. The results for B1900+Hf indicate that material constants related to hardening and thermal recovery can be obtained on the basis of such a procedure. Strain aging, thermal recovery, and unexpected material variations, however, preluded an accurate determination of the strain-rate sensitivity parameter is this exercise. The effects of casting grain size on the constitutive behavior of B1900+Hf were studied and no particular grain size effect was observed. A systematic procedure was also developed for determining the material constants in the Bodner-Partom model. Both the new test procedure and the method for determining material constants were applied to the alternate material, Mar-M247 . Test data including tensile, creep, cyclic and nonproportional biaxial (tension/torsion) loading were collected. Good correlations were obtained between the Bodner-Partom model and experiments. A literature survey was conducted to assess the effects of thermal history on the constitutive behavior of metals. Thermal history effects are expected to be present at temperature regimes where strain aging and change of microstructure are important. Possible modifications to the Bodner-Partom model to account for these effects are outlined. The use of a unified constitutive model for hot section component analyses was demonstrated by applying the Walker model and the MARC finite-element code to a B1900+Hf airfoil problem.

Chan, Kwai S.; Lindholm, Ulric S.; Bodner, S. R.; Hill, Jeff T.; Weber, R. M.; Meyer, T. G.

1986-01-01

350

Microarray analysis of ncRNA expression patterns in Caenorhabditis elegans after RNAi against snoRNA associated proteins  

Microsoft Academic Search

BACKGROUND: Short non-coding RNAs (ncRNAs) perform their cellular functions in ribonucleoprotein (RNP) complexes, which are also essential for maintaining the stability of the ncRNAs. Depletion of individual protein components of non-coding ribonucleoprotein (ncRNP) particles by RNA interference (RNAi) may therefore affect expression levels of the corresponding ncRNA, and depletion of candidate associated proteins may constitute an alternative strategy when investigating

Muhammad Nauman Aftab; Housheng He; Geir Skogerbř; Runsheng Chen

2008-01-01

351

Cosmic Microwave Background Theory  

Microsoft Academic Search

A long-standing goal of theorists has been to constrain cosmological parameters that define the structure formation theory from cosmic microwave background (CMB) anisotropy experiments and large-scale structure (LSS) observations. The status and future promise of this enterprise is described. Current band-powers in ell -space are consistent with a Delta T flat in frequency and broadly follow inflation-based expectations. That the

J. Richard Bond

1998-01-01

352

The cosmic microwave background  

NASA Technical Reports Server (NTRS)

Recent observational and theoretical investigations of the cosmic microwave background radiation (CMBR) are reviewed. Particular attention is given to spectral distortions and CMBR temperature anisotropies at large, intermediate, and small angular scales. The implications of the observations for inflationary cosmological models with curvature fluctuation are explored, and it is shown that the limits determined for intermediate-scale CMBR anisotropy almost rule out a baryon-dominated cosmology.

Silk, Joseph

1989-01-01

353

Space shuttle propellant constitutive law verification tests  

NASA Technical Reports Server (NTRS)

As part of the Propellants Task (Task 2.0) on the Solid Propulsion Integrity Program (SPIP), a database of material properties was generated for the Space Shuttle Redesigned Solid Rocket Motor (RSRM) PBAN-based propellant. A parallel effort on the Propellants Task was the generation of an improved constitutive theory for the PBAN propellant suitable for use in a finite element analysis (FEA) of the RSRM. The outcome of an analysis with the improved constitutive theory would be more reliable prediction of structural margins of safety. The work described in this report was performed by Materials Laboratory personnel at Thiokol Corporation/Huntsville Division under NASA contract NAS8-39619, Mod. 3. The report documents the test procedures for the refinement and verification tests for the improved Space Shuttle RSRM propellant material model, and summarizes the resulting test data. TP-H1148 propellant obtained from mix E660411 (manufactured February 1989) which had experienced ambient igloo storage in Huntsville, Alabama since January 1990, was used for these tests.

Thompson, James R.

1995-01-01

354

Constitutive modeling for isotropic materials (HOST)  

NASA Technical Reports Server (NTRS)

This report presents the results of the second year of work on a problem which is part of the NASA HOST Program. Its goals are: (1) to develop and validate unified constitutive models for isotropic materials, and (2) to demonstrate their usefulness for structural analyses of hot section components of gas turbine engines. The unified models selected for development and evaluation are that of Bodner-Partom and Walker. For model evaluation purposes, a large constitutive data base is generated for a B1900 + Hf alloy by performing uniaxial tensile, creep, cyclic, stress relation, and thermomechanical fatigue (TMF) tests as well as biaxial (tension/torsion) tests under proportional and nonproportional loading over a wide range of strain rates and temperatures. Systematic approaches for evaluating material constants from a small subset of the data base are developed. Correlations of the uniaxial and biaxial tests data with the theories of Bodner-Partom and Walker are performed to establish the accuracy, range of applicability, and integability of the models. Both models are implemented in the MARC finite element computer code and used for TMF analyses. Benchmark notch round experiments are conducted and the results compared with finite-element analyses using the MARC code and the Walker model.

Lindholm, U. S.; Chan, K. S.; Bodner, S. R.; Weber, R. M.; Walker, K. P.; Cassenti, B. N.

1985-01-01

355

Constitutional ring chromosomes and tumour suppressor genes.  

PubMed Central

The types of malignancy reported in carriers of constitutional ring chromosomes r(11), r(13), and r(22) are concordant with the chromosomal assignment of tumour suppressor loci associated with Wilms' tumour, retinoblastoma, and meningioma. It is suggested that the somatic instability of ring chromosomes may play a role in this association and that constitutional ring chromosomes may be a source for mapping of tumour suppressor loci with the potential for covering most or all of the human genome. The hypothesis predicts the presence of a locus on chromosome 10 associated with follicular carcinoma of the thyroid, in line with previous cytogenetic findings of rearrangements involving chromosome 10 in thyroid tumours, and a locus on chromosome 22 associated with testicular cancer. Development of neurofibromatoses (NF) that do not fulfil the clinical criteria of neurofibromatosis type 2 (NF2) in carriers with r(22) suggests either the presence of an additional NF locus on chromosome 22 or that ring chromosome mediated predisposition to somatic mutation of a specific tumour suppressor may be associated with atypical development of features usually associated with germline mutations. PMID:1336057

Tommerup, N; Lothe, R

1992-01-01

356

Computer simulation of the mathematical modeling involved in constitutive equation development: Via symbolic computations  

NASA Technical Reports Server (NTRS)

Development of new material models for describing the high temperature constitutive behavior of real materials represents an important area of research in engineering disciplines. 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 process can be of significant benefit. A computerized procedure (SDICE) capable of efficiently deriving potential based constitutive models, in analytical form is presented. This package, running under MACSYMA, has the following features: partial differentiation, tensor computations, automatic grouping and labeling of common factors, expression substitution and simplification, back substitution of invariant and tensorial relations and a relational data base. Also limited aspects of invariant theory were incorporated into SDICE due to the utilization of potentials as a starting point and the desire for these potentials to be frame invariant (objective). Finally not only calculation of flow and/or evolutionary laws were accomplished but also the determination of history independent nonphysical coefficients in terms of physically measurable parameters, e.g., Young's modulus, was achieved. The uniqueness of SDICE resides in its ability to manipulate expressions in a general yet predefined order and simplify expressions so as to limit expression growth. Results are displayed when applicable utilizing index notation.

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

1989-01-01

357

Granular materials: constitutive equations and strain localization  

NASA Astrophysics Data System (ADS)

Strain localization into shear bands is commonly observed in natural soil masses, as well as in human-built embankments, footings, retaining walls and other geotechnical structures. Numerical predictions for the process of shear band formation are critically dependent on the constitutive equations employed. In this paper, the plane strain "double-shearing" constitutive model (e.g., Spencer, A.J.M., 1964. A theory of the kinematics of ideal soils under plane strain conditions. Journal of the Mechanics and Physics of Solids 12, 337-351; Spencer, A.J.M., 1982, Deformation of ideal granular materials. In: Hopkins, H.G., Sewell, M.J. (Eds.), Mechanics of Solids. Pergamon Press, Oxford and New York, pp. 607-652; Mehrabadi, M.M., Cowin, S.C., 1978. Initial planar deformation of dilatant granular materials. Journal of the Mechanics and Physics of Solids 26, 269-284; Nemat-Nasser, S., Mehrabadi, M.M., Iwakuma, T. 1981. On certain macroscopic and microscopic aspects of plastic flow of ductile materials. In: Nemat-Nasser, S. (Ed.), Three-dimensional Constitutive Relations and Ductile Fracture. North-Holland, Amsterdam, pp. 157-172; Anand, L., 1983. Plane deformations of ideal granular materials. Journal of the Mechanics and Physics of Solids 31, 105-122) is generalized to three dimensions including the effects of elastic deformation and pre-peak behavior. The constitutive model is implemented in a finite element program and is used to predict the formation of shear bands in plane strain compression, and plane strain cylindrical cavity expansion. The predictions from the model are shown to be in good quantitative agreement with the recent experiments of Han, C., Drescher, A., (1993. Shear bands in biaxial tests on dry coarse sand. Soils and Foundations 33, 118-132) and Alsiny, H., Vardoulakis, I., Drescher, A., (1992. Deformation localization in cavity inflation experiments on dry sand. Geotechnique 42, 395-410) on a dry sand. The constitutive model is also used to predict the stress state in a static sand pile — a topic which has occupied the attention of many investigators in recent years. In our simulations we model an initially loose sand mass as a cohesionless material with a mobilized internal friction coefficient which evolves from an initial value of zero to a saturation value. The formation of a sand pile is numerically modeled as a two-step process: (i) In the first step a conical sand mass is placed between a flat rigid surface and an axi-symmetric conical mold. Interaction between the sand mass and the rigid base plate is modeled using an interface friction coefficient which has the same value as the saturation value of the internal friction coefficient. The sand mass (which is confined between the base plate and the conical mold) is subjected to gravity loading, and the system is allowed to equilibrate. (ii) In the second step the conical mold is quickly lifted and the sand mass allowed to reach a new equilibrate, but slightly slumped configuration. In the process of slight slumping, there is non-homogeneous plastic deformation of the sand pile. This non-homogeneous plastic deformation, coupled with the evolving internal friction coefficient, naturally gives rise to a static stress state which exhibits the interesting feature that the vertical stress distribution at the base of the sand pile does not have a maximum under the apex of the cone, but shows a local dip there.

Anand, L.; Gu, C.

2000-08-01

358

Targets, backgrounds, and discrimination  

NASA Astrophysics Data System (ADS)

The present volume discusses a model-based aircraft identification technique, target intensity and angle scintillations, spatiotemporal nonstationary scene generation, an overview of the Strategic Scene Generation Model (SSGM), nuclear backgrounds for SSGM, and an atmospheric and transmittance code for 50-300 km altitudes. Also discussed are a data base for airborne target signatures, the auroral module of the Strategic High Altitude Radiance Code, and the 3D characteristics of underexpanded and overexpanded rectangular jets. (No individual items are abstracted in this volume)

Accetta, J. S.; Kelley, G. H.

359

Model-based target and background characterization  

NASA Astrophysics Data System (ADS)

Up to now most approaches of target and background characterization (and exploitation) concentrate solely on the information given by pixels. In many cases this is a complex and unprofitable task. During the development of automatic exploitation algorithms the main goal is the optimization of certain performance parameters. These parameters are measured during test runs while applying one algorithm with one parameter set to images that constitute of image domains with very different domain characteristics (targets and various types of background clutter). Model based geocoding and registration approaches provide means for utilizing the information stored in GIS (Geographical Information Systems). The geographical information stored in the various GIS layers can define ROE (Regions of Expectations) and may allow for dedicated algorithm parametrization and development. ROI (Region of Interest) detection algorithms (in most cases MMO (Man- Made Object) detection) use implicit target and/or background models. The detection algorithms of ROIs utilize gradient direction models that have to be matched with transformed image domain data. In most cases simple threshold calculations on the match results discriminate target object signatures from the background. The geocoding approaches extract line-like structures (street signatures) from the image domain and match the graph constellation against a vector model extracted from a GIS (Geographical Information System) data base. Apart from geo-coding the algorithms can be also used for image-to-image registration (multi sensor and data fusion) and may be used for creation and validation of geographical maps.

Mueller, Markus; Krueger, Wolfgang; Heinze, Norbert

2000-07-01

360

A continuum constitutive model for amorphous metallic materials  

E-print Network

A finite-deformation, Coulomb-Mohr type constitutive theory for the elastic-viscoplastic response of pressure-sensitive and plastically-dilatant isotropic materials has been developed. The constitutive model has been ...

Su, Cheng, Ph. D. Massachusetts Institute of Technology

2007-01-01

361

Making American: Constitutive Rhetoric in the Cold War  

E-print Network

issue of Playboy. Each chapter is designed to provide an example of what a constitutive analysis in the style of White would look like. The project begins with a description of the theories and analyses, including constitutive rhetoric, postmodernism...

Thorpe, Martha

2011-10-21

362

Constitutional rights versus malpractice insurance settlements.  

PubMed

The increasing costs and complexity of malpractice litigation have created an statutory right that allows malpractice insurance companies to settle malpractice claims regardless of the desires of the defendant physician. In the past, the consequences of settling a malpractice claim out of court were not as important as they are today. The Health Care Quality Improvement Act of 1986 mandates that any settlement in behalf of a physician be documented in the National Practitioner Data Bank (NPDB), which must be consulted every time the physician is credentialed. This NPDB requirement denies due process to health care providers and thus becomes a violation of the federal and many state constitutions. Physician executives and medical leaders must bring these issues to the table and negotiate solutions before damage to practicing physicians and the U.S. health care delivery system caused by this legal paradox become too severe. PMID:10139374

Trentalance, A E

1994-08-01

363

Rotational and constitutional dynamics of caged supramolecules  

PubMed Central

The confinement of molecular species in nanoscale environments leads to intriguing dynamic phenomena. Notably, the organization and rotational motions of individual molecules were controlled by carefully designed, fully supramolecular host architectures. Here we use an open 2D coordination network on a smooth metal surface to steer the self-assembly of discrete trimeric guest units, identified as noncovalently bound dynamers. Each caged chiral supramolecule performs concerted, chirality-preserving rotary motions within the template honeycomb pore, which are visualized and quantitatively analyzed using temperature-controlled scanning tunneling microscopy. Furthermore, with higher thermal energies, a constitutional system dynamics appears, which is revealed by monitoring repetitive switching events of the confined supramolecules’ chirality signature, reflecting decay and reassembly of the caged units. PMID:21098303

Kühne, Dirk; Klappenberger, Florian; Krenner, Wolfgang; Klyatskaya, Svetlana; Ruben, Mario; Barth, Johannes V.

2010-01-01

364

Backgrounds Data Center  

NASA Astrophysics Data System (ADS)

The Backgrounds Data Center (BDC) is the designated archive for backgrounds data collected by Ballistic Missile Defense Organization (BMDO) programs, some of which include ultraviolet sensors. Currently, the BDC holds ultraviolet data from the IBSS, UVPI, UVLIM, and FUVCAM sensors. The BDC will also be the prime archive for Midcourse Space Experiment (MSX) data and is prepared to negotiate with program managers to handle other datasets. The purpose of the BDC is to make data accessible to users and to assist them in analyzing it. The BDC maintains the Science Catalog Information Exchange System (SCIES) allowing remote users to log in, read or post notices about current programs, search the catalogs for datasets of interest, and submit orders for data. On-site facilities are also available for the analysis of data, and consist of VMS and UNIX workstations with access to software analysis packages such as IDL, IRAF, and Khoros. Either on-site or remotely, users can employ the BDC-developed graphical user interface called the Visual Interface for Space and Terrestrial Analysis (VISTA) to generate catalog queries and to display and analyze data. SCIES and VISTA permit nearly complete access to BDC services and capabilities without the need to be physically present at the data center.

Snyder, William A.; Gursky, Herbert; Heckathorn, Harry M.; Lucke, Bob L.; Dorland, Bryan N.; Kessel, R. A.; Berg, S. L.; Dombrowski, E. G.

1994-09-01

365

Freedom's Law: The Moral Reading of the American Constitution  

E-print Network

of the Constitution. The United States Consti­ tution, like most contemporary constitutions, defines individual rights in very broad and abstract language. The Fourth Amendment, for example, protects against "unreasonable searches and seizures." The Eighth Amend... is constitutional person, then it is protected by the Equal Protection Clause of the Fourteenth Amend­ ment, which declares that no state may deny to any person equal protec­ tion of the laws. Given that Dworkin argues that the language in the Constitution should...

Himma, Kenneth Einar

366

Pinning down the Constitution: Interactively Teaching Congress's Power, Federalism, and Constitutional Interpretation  

ERIC Educational Resources Information Center

"Pin-the-Tail-on-the-Constitution" is an engaged-learning activity that has been conducted in 26 classes over the past four years. The activity teaches multiple themes commonly included in a variety of courses on American politics such as federalism, congressional powers, the role of the federal courts, and the relevance of the commerce…

Emenaker, Ryan

2014-01-01

367

Inhibition of a background potassium channel by Gq protein alpha-subunits.  

PubMed

Two-pore-domain K(+) channels provide neuronal background currents that establish resting membrane potential and input resistance; their modulation provides a prevalent mechanism for regulating cellular excitability. The so-called TASK channel subunits (TASK-1 and TASK-3) are widely expressed, and they are robustly inhibited by receptors that signal through Galphaq family proteins. Here, we manipulated G protein expression and membrane phosphatidylinositol 4,5-bisphosphate (PIP(2)) levels in intact and cell-free systems to provide electrophysiological and biochemical evidence that inhibition of TASK channels by Galphaq-linked receptors proceeds unabated in the absence of phospholipase C (PLC) activity, and instead involves association of activated Galphaq subunits with the channels. Receptor-mediated inhibition of TASK channels was faster and less sensitive to a PLCbeta1-ct minigene construct than inhibition of PIP(2)-sensitive Kir3.4(S143T) homomeric channels that is known to be dependent on PLC. TASK channels were strongly inhibited by constitutively active Galphaq, even by a mutated version that is deficient in PLC activation. Receptor-mediated TASK channel inhibition required exogenous Galphaq expression in fibroblasts derived from Galphaq/11 knockout mice, but proceeded unabated in a cell line in which PIP(2) levels were reduced by regulated overexpression of a lipid phosphatase. Direct application of activated Galphaq, but not other G protein subunits, inhibited TASK channels in excised patches, and constitutively active Galphaq subunits were selectively coimmunoprecipitated with TASK channels. These data indicate that receptor-mediated TASK channel inhibition is independent of PIP(2) depletion, and they suggest a mechanism whereby channel modulation by Galphaq occurs through direct interaction with the ion channel or a closely associated intermediary. PMID:16492788

Chen, Xiangdong; Talley, Edmund M; Patel, Nitin; Gomis, Ana; McIntire, William E; Dong, Biwei; Viana, Félix; Garrison, James C; Bayliss, Douglas A

2006-02-28

368

Resolution and Letter to Congress from the Constitutional Convention  

ERIC Educational Resources Information Center

This article explores the drafting of the United States Constitution. The United States Constitution is the longest-lasting written national constitution in the world. Its four parchment pages serve as the blueprint for a government under which more than 290 million Americans live. It holds great significance for the American people and for others…

Potter, Lee Ann

2005-01-01

369

Constitutional Provisions on the Press: A World View.  

ERIC Educational Resources Information Center

A study examined the legal treatment of the press in constitutions or other basic legal institutional documents from around the world. Sixty-three constitutions or basic documents from the Western World, the Communist Bloc, the Middle East, Africa, Southeast Asia, and Latin America were analyzed. Analysis revealed that most constitutions open with…

Paraschos, Manny

370

Frederick Douglass Changed My Mind about the Constitution  

ERIC Educational Resources Information Center

Frederick Douglass dramatically and publicly changed his own mind about the Constitution. Like Frederick Douglass, the author had originally viewed the Constitution as pro-slavery. Yet a close look at Douglass's writings revealed a Constitution that empowered the federal government to abolish slavery.

Oakes, James

2008-01-01

371

Teaching United States History with a Constitutional Focus.  

ERIC Educational Resources Information Center

The basic premise of the paper is that the Constitution should be the main theme of courses in United States history at grade 8. Students should study a unit on the Constitution, as is commonly done at that level. In addition, other units taught throughout the school year should emphasize constitutional principles, values, and issues. Three…

Hyland, John T.

372

The Separation of Church and State. Exploring the Constitution Series.  

ERIC Educational Resources Information Center

This textbook on the separation of church and state continues the "Exploring the Constitution Series," which introduces important areas of constitutional law. Intended to serve either as a reference work, a supplement to a standard textbook, or as the textbook for a course, this volume covers the constitutional issues of prayer in public schools,…

McWhirter, Darien A.

373

Constitutive modeling of multiphase flows with moving interfaces and contact line  

NASA Astrophysics Data System (ADS)

A continuum description of multiphase flows, in which excess physical quantities associated with phase interfaces and the three-phase contact line are incorporated, is briefly presented. A thermodynamic analysis, based on the Müller-Liu thermodynamic approach of the second law of thermodynamics, is performed to derive the expressions of the constitutive variables in thermodynamic equilibrium. Non-equilibrium responses are proposed by use of a quasi-linear theory. A set of constitutive equations for the surface and line constitutive quantities is postulated. Some restrictions for the emerging material parameters are derived by means of the minimum conditions of the surface and line entropy productions in thermodynamic equilibrium. Hence, a complete continuum mechanical model to describe excess surface and line physical quantities is formulated.

Wang, Yongqi; Oberlack, Martin; Zieleniewicz, Andreas

2013-11-01

374

On the treatment of non-solvable implicit constitutive relations in solid mechanics  

NASA Astrophysics Data System (ADS)

We report results concerning the treatment of elastic bodies described by implicit constitutive relations which are not solvable, in the sense of expressing the stresses as functions of the strains or vice versa. Motivated by the theory of generalized hyperelastic materials, which applies also to polyatomic crystals, the field equations to be solved for a body described by a non-solvable implicit constitutive relation in the above sense are lied down. In addition to the momentum equation, auxiliary variables come into play. These variables are accompanied by an equation governing them. We specialize to the case of an isotropic constitutive relation and give some conditions in order for a broad subclass of an isotropic body to be a generalized material. Then, we lay down the field equations for such a body.

Sfyris, D.; Bustamante, R.

2014-05-01

375

A Comparative Analysis of Constitutive Promoters Located in Adeno-Associated Viral Vectors  

PubMed Central

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 ?-actin, human elongation factor-1?, chicken ?-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 ?-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. PMID:25170953

Ota, Akinobu; Hossain, Ekhtear; Konishi, Yuko; Hosokawa, Yoshitaka; Konishi, Hiroyuki

2014-01-01

376

A comparative analysis of constitutive promoters located in adeno-associated viral vectors.  

PubMed

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 ?-actin, human elongation factor-1?, chicken ?-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 ?-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. PMID:25170953

Damdindorj, Lkhagvasuren; Karnan, Sivasundaram; Ota, Akinobu; Hossain, Ekhtear; Konishi, Yuko; Hosokawa, Yoshitaka; Konishi, Hiroyuki

2014-01-01

377

Holography for Schrodinger backgrounds  

E-print Network

We discuss holography for Schrodinger solutions of both topologically massive gravity in three dimensions and massive vector theories in (d+1) dimensions. In both cases the dual field theory can be viewed as a d-dimensional conformal field theory (two dimensional in the case of TMG) deformed by certain operators that respect the Schrodinger symmetry. These operators are irrelevant from the viewpoint of the relativistic conformal group but they are exactly marginal with respect to the non-relativistic conformal group. The spectrum of linear fluctuations around the background solutions corresponds to operators that are labeled by their scaling dimension and the lightcone momentum k_v. We set up the holographic dictionary and compute 2-point functions of these operators both holographically and in field theory using conformal perturbation theory and find agreement. The counterterms needed for holographic renormalization are non-local in the v lightcone direction.

Monica Guica; Kostas Skenderis; Marika Taylor; Balt van Rees

2010-08-11

378

The diffuse UV background  

NASA Technical Reports Server (NTRS)

The diffuse radiation field in the UV (900-3,000 A) affects the structure of galactic molecular clouds and conveys important information concerning the physical characteristics and spatial distribution of gas and dust in the universe. Continuum emission in this range is probably dominated by interstellar dust scattering in our galaxy. For view directions and angular resolutions allowing observations in the rifts between galactic dust clouds, the background due to the integrated light of spiral galaxies may be detected, providing important information on their structure and evolution. The redshifted emission from an intergalactic medium may be observable in the regions between nearby bright galaxies. Present observations provide weak constraints on the radiation field required to ionize the intergalactic medium at the level required by the Gunn-Peterson test.

Paresce, F.; Jakobsen, P.

1980-01-01

379

Free Press in a Constitutional Democracy  

ERIC Educational Resources Information Center

This article describes a lesson in which students will examine several views expressed by the founders to understand the context for including freedom of the press in the First Amendment. Students will be asked to think about the role that the news media and the need to be an informed citizen continue to play in our democracy. Students will…

Lucianek, Christine

2014-01-01

380

Redressing Deprivations of Rights Secured by State Constitutions Outside the Shadow of the Supreme Court's Constitutional Remedies Jurisprudence  

Microsoft Academic Search

The second generation of state constitutionalism is now emerging. With the methodology of autonomous state constitutional protection more clearly defined, courts and legislatures are turning to the task of determining when, and from whom, they should award damages to citizens deprived of their state constitutional rights. State courts, as well as legislatures contemplating statutes authorizing damage actions, will be tempted

Gary S. Gildin

2011-01-01

381

The Constitution as Based on the Consent of the Governed—Or, Should We Have an Unwritten Constitution?  

Microsoft Academic Search

It is useful to embrace continuity in describing basic differences we have in giving effect to the Constitution, especially if particular ways of communicating help us convey and understand what is at stake. The individual who originated the term “non-interpretivist” to describe judicial review implementing the unwritten constitution, for example, continues to believe that the best approach to constitutional interpretation

Thomas B. McAffee

2001-01-01

382

Constitutive Relation in Transitional Granular Flows  

NASA Technical Reports Server (NTRS)

To study the constitutive behavior of granular materials, the presence of gravity is detrimental. Although empirical relations have been obtained for engineering designs to control granular flows on Earth, it is not known how well these Earthbound relations can be used in another gravity field. Fundamental understanding must be derived to reliably design for granular flows in space exploration. There are two extremes of granular flows of which significant amount of knowledge is available. One deals with a dense and quasi-static situation where the deformation rate nearly vanishes. The other deals with dilute and rapidly fluctuating grain velocities where particle inertia dominates. This project, funded by the NASA Microgravity Fluid Physics Program, aims to study this transitional regime via physical experiments and computer simulations. A conceptual model has been established as described below. There are two natural time scales in a granular flow. One is the travel time between two consecutive collisions and the other is the duration of a collision contact. At a very low shear-rate, the shear-induced particle velocity is low. Hence the travel time between collisions is longer than the contact time between colliding particles. Binary collisions prevail. As the shear-rate increases, the traveling time between collisions reduces and the probability of multiple collisions goes up. These particle groups disperse shortly after and new groups form. When shear-rate is further increased, clusters grow in size due to an increasing chance for free particles to join before groups have the time to disperse. The maximum cluster size may depend on the global concentration and material properties. As the solid concentration approaches zero, the cluster size goes to one particle diameter. The maximum possible cluster size under any condition is the container size, provided that the shear flow is inside a container. The critical shear-rates that dictate the initiation of the multiple contacts, and the size and lifetime of the collision clusters, are functions of the concentration also. A 'regime' theory has been proposed by Babic et al. This theory suggested that both the solid concentration, C, and the non-dimensional shear-rate, B, are important in determining the regimes of the granular constitutive law. Additional information is included in the original extended abstract.

Shen, Hayley H.; Hanes, Daniel M.; Jenkins, James T.

2002-01-01

383

Constitutive Theory Developed for Monolithic Ceramic Materials  

NASA Technical Reports Server (NTRS)

With the increasing use of advanced ceramic materials in high-temperature structural applications such as advanced heat engine components, the need arises to accurately predict thermomechanical behavior that is inherently time-dependent and that is hereditary in the sense that the current behavior depends not only on current conditions but also on the material's thermomechanical history. Most current analytical life prediction methods for both subcritical crack growth and creep models use elastic stress fields to predict the time-dependent reliability response of components subjected to elevated service temperatures. Inelastic response at high temperatures has been well documented in the materials science literature for these material systems, but this issue has been ignored by the engineering design community. From a design engineer's perspective, it is imperative to emphasize that accurate predictions of time-dependent reliability demand accurate stress field information. Ceramic materials exhibit different time-dependent behavior in tension and compression. Thus, inelastic deformation models for ceramics must be constructed in a fashion that admits both sensitivity to hydrostatic stress and differing behavior in tension and compression. A number of constitutive theories for materials that exhibit sensitivity to the hydrostatic component of stress have been proposed that characterize deformation using time-independent classical plasticity as a foundation. However, none of these theories allow different behavior in tension and compression. In addition, these theories are somewhat lacking in that they are unable to capture the creep, relaxation, and rate-sensitive phenomena exhibited by ceramic materials at high temperatures. The objective of this effort at the NASA Lewis Research Center has been to formulate a macroscopic continuum theory that captures these time-dependent phenomena. Specifically, the effort has focused on inelastic deformation behavior associated with these service conditions by developing a multiaxial viscoplastic constitutive model that accounts for time-dependent hereditary material deformation (such as creep and stress relaxation) in monolithic structural ceramics. Using continuum principles of engineering mechanics, we derived the complete viscoplastic theory from a scalar dissipative potential function.

Janosik, Lesley A.

1998-01-01

384

Elastic-Plastic Constitutive Equation of WC-Co Cemented Carbides with Anisotropic Damage  

NASA Astrophysics Data System (ADS)

Elastic-plastic constitutive equation of WC-Co cemented carbides with anisotropic damage is proposed to predict a precise service life of cold forging tools. A 2nd rank symmetric tensor damage tensor is introduced in order to express the stress unilaterality; a salient difference in uniaxial behavior between tension and compression. The conventional framework of irreversible thermodynamics is used to derive the constitutive equation. The Gibbs potential is formulated as a function of stress, damage tensor, isotropic hardening variable and kinematic hardening variable. The elastic-damage constitutive equation, conjugate forces of damage, isotropic hardening and kinematic hardening variable is derived from the potential. For the kinematic hardening variable, the superposition of three kinematic hardening laws is employed in order to improve the cyclic behavior of the material. For the evolution equation of the damage tensor, the damage is assumed to progress by fracture of the Co matrix — WC particle interface and by the mechanism of fatigue, i.e. the accumulation of microscopic plastic strain in matrix and particles. By using the constitutive equations, calculation of uniaxial tensile and compressive test is performed and the results are compared with the experimental ones in the literature. Furthermore, finite element analysis on cold forward extrusion was carried out, in which the proposed constitutive equation was employed as die insert material.

Hayakawa, Kunio; Nakamura, Tamotsu; Tanaka, Shigekazu

2007-05-01

385

Elastic-Plastic Constitutive Equation of WC-Co Cemented Carbides with Anisotropic Damage  

SciTech Connect

Elastic-plastic constitutive equation of WC-Co cemented carbides with anisotropic damage is proposed to predict a precise service life of cold forging tools. A 2nd rank symmetric tensor damage tensor is introduced in order to express the stress unilaterality; a salient difference in uniaxial behavior between tension and compression. The conventional framework of irreversible thermodynamics is used to derive the constitutive equation. The Gibbs potential is formulated as a function of stress, damage tensor, isotropic hardening variable and kinematic hardening variable. The elastic-damage constitutive equation, conjugate forces of damage, isotropic hardening and kinematic hardening variable is derived from the potential. For the kinematic hardening variable, the superposition of three kinematic hardening laws is employed in order to improve the cyclic behavior of the material. For the evolution equation of the damage tensor, the damage is assumed to progress by fracture of the Co matrix - WC particle interface and by the mechanism of fatigue, i.e. the accumulation of microscopic plastic strain in matrix and particles. By using the constitutive equations, calculation of uniaxial tensile and compressive test is performed and the results are compared with the experimental ones in the literature. Furthermore, finite element analysis on cold forward extrusion was carried out, in which the proposed constitutive equation was employed as die insert material.

Hayakawa, Kunio; Nakamura, Tamotsu; Tanaka, Shigekazu [Department of Mechanical Engineering, Shizuoka University, 3-5-1 Johoku, Hamamatsu 432-8561 (Japan)

2007-05-17

386

Background sources in optical communications  

NASA Technical Reports Server (NTRS)

The characterization and measurement of background radiation relevant to optical communications system performance is addressed. The necessary optical receiver parameters are described, and radiometric concepts required for the calculation of collected background power are developed. The most important components of optical background power are discussed, and their contribution to the total collected background power in various communications scenarios is examined.

Vilnrotter, V. A.

1983-01-01

387

Cosmic Microwave Background  

NASA Astrophysics Data System (ADS)

The cosmic microwave background (CMB) radiation, the relic of the early phases of the expanding universe, is bright, full of information, and difficult to measure. Along with the recession of galaxies and the primordial nucleosynthesis, it is one of the strongest signs that the Hot Big Bang Model of the universe is correct. It is brightest around 2 mm wavelength, has a temperature of T_{cmb} = 2.72548 ± 0.00057 K, and has a blackbody spectrum within 50 parts per million. Its spatial fluctuations (around 0.01% on 1{}^{circ } scales) are possibly the relics of quantum mechanical processes in the early universe, modified by processes up to the decoupling at a redshift of about 1,000 (when the primordial plasma became mostly transparent). In the cold dark matter (DM) model with cosmic acceleration (? CDM), the fluctuation statistics are consistent with the model of inflation and can be used to determine other parameters within a few percent, including the Hubble constant, the ? constant, the densities of baryonic and dark matter, and the primordial fluctuation amplitude and power spectrum slope. In addition, the polarization of the fluctuations reveals the epoch of reionization at a redshift approximately twice that determined from the Gunn-Peterson trough due to optically thick Lyman ? absorption in QSO spectra. It is of historic importance, and a testament to the unity of theory and experiment, that we now have a standard model of cosmology that is consistent with all of the observations.Current observational challenges include (1) improvement of the spectrum distortion measurements, especially at long wavelengths, where the measured background is unexpectedly bright; (2) the search for the B-mode polarization (the divergence-free part of the polarization map), arising from propagating gravitational waves; and (3) the extension of fluctuation measurements to smaller angular scales. Much more precise spectrum observations near 2 mm are likely and would test some very interesting theories. Current theoretical challenges include explanation of the dark matter and dark energy; understanding, estimating, and removing the interference of foreground sources that limit the measurements of the CMB; detailed understanding of the influence of nonequilibrium processes on the decoupling and reionization phases; and searches for signs of the second order or exotic processes (e.g., isocurvature fluctuations, cosmic strings, non-Gaussian fluctuations). At this writing, we await the cosmological results of the Planck mission.

Mather, John; Hinshaw, Gary; Page, Lyman

388

Health Care for Micronesians and Constitutional Rights  

PubMed Central

Under the Compacts of Free Association (COFA), people from the Freely Associated States — the Republic of Palau (ROP), the Republic of the Marshall Islands (RMI), and the Federated States of Micronesia (FSM) — have been migrating to the United States in increasing numbers. In 1996, Congress passed broad welfare reform (Personal Responsibility and Work Opportunity Reconciliation Act) which limited certain federal benefits previously available to COFA migrants, including Medicaid benefits. Prior to July 2010, the State of Hawai‘i had continued to include COFA migrants under its state-funded Medicaid program. In the face of budget constraints, the State removed these people from its Medicaid rolls. A challenge on the legal basis of the denial of equal protection of the laws, ie, the Fourteenth Amendment to the US Constitution, was successful in reinstating health care to the COFA migrants in December 2010. From the health worker's perspective, regardless of various social justice arguments that may have been marshaled in favor of delivering health care to the people, it was an appeal to the judicial system that succeeded. From the attorney's perspective, the legal victories are potentially limited to the four walls of the courtroom without community involvement and related social justice movements. Together, the authors propose that in order to better address the issue of health care access for Micronesian peoples, we must work together, as health and legal advocates, to define a more robust vision of both systems that includes reconciliation and community engagement. PMID:22235150

Shek, Dina

2011-01-01

389

Constitutive exposure of phosphatidylserine on viable cells  

PubMed Central

Apoptotic cells are quickly recognized and engulfed by phagocytes to prevent the release of noxious materials from dying cells. Phosphatidylserine (PS) exposed on the surface of apoptotic cells is a proposed “eat-me” signal for the phagocytes. Transmembrane protein 16F (TMEM16F), a membrane protein with eight transmembrane segments, has the Ca-dependent phospholipid scramblase activity. Here we show that when lymphoma cells were transformed with a constitutively active form of TMEM16F, they exposed a high level of PS that was comparable to that observed on apoptotic cells. The PS-exposing cells were morphologically normal and grew normally. They efficiently responded to interleukin 3 and underwent apoptosis upon treatment with Fas ligand. The viable PS-exposing cells bound to peritoneal macrophages at 4 °C, but not at 25 °C. Accordingly, these cells were not engulfed by macrophages. When apoptotic cells were injected i.v. into mice, they were phagocytosed by CD11c+CD8+ dendritic cells (DCs) in the spleen, but the PS-exposing living cells were not phagocytosed by these DCs. Furthermore, when PS-exposing lymphoma cells were transplanted s.c. into nude mice, they generated tumors as efficiently as parental lymphoma cells that did not expose PS. These results indicated that PS exposure alone is not sufficient to be recognized by macrophages as an eat-me signal. PMID:22084121

Segawa, Katsumori; Suzuki, Jun; Nagata, Shigekazu

2011-01-01

390

Constitutive Modeling of Magnesium Alloy Sheets  

SciTech Connect

Magnesium alloy sheets have unique mechanical properties: high in-plane anisotropy/asymmetry of yield stress and hardening response, which have not been thoroughly studied. The unusual mechanical behavior of magnesium alloys has been understood by the limited symmetry crystal structure of h.c.p metals and thus by deformation twinning. In this paper, the phenomenological continuum plasticity models considering the unusual plastic behavior of magnesium alloy sheet were developed for a finite element analysis. A new hardening law based on two-surface model was developed to consider the general stress-strain response of metal sheets such as Bauschinger effect, transient behavior and the unusual asymmetry. Three deformation modes observed during the continuous tension/compression tests were mathematically formulated with simplified relations between the state of deformation and their histories. In terms of the anisotropy and asymmetry of the initial yield stress, the Drucker-Prager's pressure dependent yield surface was modified to include the anisotropy of magnesium alloys. Also, characterization procedures of material parameters for the constitutive equations were presented and finally the correlation of simulation with measurements was performed to validate the proposed theory.

Lee, M. G.; Piao, K.; Wagoner, R. H. [Department of Materials Science and Engineering, 2041 College Road, Ohio State University, Columbus, OH 43210 (United States); Lee, J. K. [Department of Mechanical Engineering, Scott Laboratory, 201 West 19th Avenue, Ohio State University, Columbus, OH 43210 (United States); Chung, K. [School of Materials Science and Engineering, Intelligent Textile System Research Center, Seoul National University, 56-1, Shinlim-Dong, Kwanak-Ku, Seoul 151-742 (Korea, Republic of); Kim, H. Y. [Division of Mechanical Engineering and Mechatronics, Kangwon National University, 192-1 Hyoja 2-Dong, Chunchon, Gangwon-Do, 200-701 (Korea, Republic of)

2007-05-17

391

A micromorphic model for monolayer hexagonal boron nitride with determined constitutive constants by phonon dispersions  

NASA Astrophysics Data System (ADS)

A two dimensional (2D) micromorphic model is developed for monolayer hexagonal boron nitride (h-BN). Theoretical expressions of phonon dispersions for 2D crystals are derived based on the simplified governing equations of specialized three dimensional micromorphic crystals. The constitutive constants of governing equations of the h-BN micromorphic model are determined, which is performed by fitting the available phonon dispersions data of experimental measurements and first-principles calculations with our theoretical expressions. The obtained Young's modulus and Poisson's ratio of h-BN are comparable with the results of ab initio calculations and inelastic x-ray scattering experiments, thus the constitutive relations of the h-BN model are verified, which also indicates that mechanical properties of monolayer h-BN could be characterized by our 2D micromorphic model.

Zhang, Bin; Yang, Gang

2014-10-01

392

Ordered rate constitutive theories for thermoviscoelastic solids without memory in Lagrangian description using Gibbs potential  

NASA Astrophysics Data System (ADS)

The paper presents rate constitutive theories for finite deformation of homogeneous, isotropic, compressible, and incompressible thermoviscoelastic solids without memory in Lagrangian description derived using the second law of thermodynamics expressed in terms of Gibbs potential ?. To ensure thermodynamic equilibrium during evolution, the rate constitutive theories must be derived using entropy inequality [as other three conservation and balance laws are do not provide a mechanism for deriving constitutive theories for the deforming matter (Surana in Advanced mechanics of continuua. in preparation, 2014)]. The two forms of the entropy inequality in ? derived using conjugate pairs {mathbf{?}^*} , {[dot{J}]} : first Piola-Kirchhoff stress tensor and material derivative of the Jacobian of deformation and {mathbf{?}^{[0]}} , {dot{mathbf{\\varepsilon}}_{[0]}} ; second Piola-Kirchhoff stress tensor and material derivative of Green's strain tensor are precisely equivalent as the conjugate pairs {mathbf{?}^*} , {[dot{J}]} and {mathbf{?}^{[0]}} , {dot{mathbf{\\varepsilon}}_{[0]}} are transformable from each other. In the present work, we use {mathbf{?}^{[0]}} , {dot{mathbf{\\varepsilon}}_{[0]}} as conjugate pair. Two possible choices of dependent variables in the constitutive theories: ?, ?, {mathbf{?}^{[0]}} , {mathbf{q}} and ?, ?, {mathbf{\\varepsilon}_{[0]}} , {mathbf{q}} (in which ? is entropy density and {mathbf{q}} is heat vector) are explored based on conservation and balance laws. It is shown that the choice of ?, ?, {mathbf{\\varepsilon}_{[0]}} , {mathbf{q}} is essential when the entropy inequality is expressed in terms of ?. The arguments of these dependent variables are decided based on desired physics. Viscoelastic behavior requires considerations of at least {mathbf{\\varepsilon}_{[0]}} and {dot{mathbf{\\varepsilon}}_{[0]}} (or {mathbf{\\varepsilon}_{[1]}} ) in the constitutive theories. We generalize and consider strain rates {mathbf{\\varepsilon}_{[i]}} ; i = 0, 1, …, n-1 as arguments of the dependent variables in the derivations of the ordered rate theories of up to orders n. At the onset, {mathbf{?}^{[0]}} , {mathbf{\\varepsilon}_{[i]}} ; i = 0, 1, …, n-1, ? and {mathbf{g}} are considered as arguments of ?, ?, {mathbf{\\varepsilon}_{[n]}} and {mathbf{q}} . When {dot{?}} is substituted in the entropy inequality, the resulting conditions eliminate ? as a dependent variable, reduce arguments of some of the dependent variables in the constitutive theory etc. but do not provide a mechanism to derive constitutive theories for {mathbf{\\varepsilon}_{[i]}} and {mathbf{q}} . The stress tensor {mathbf{?}^{[0]}} is decomposed into equilibrium stress {{}_e mathbf{?}^{[0]}} and deviatoric stress {{}_d mathbf{?}^{[0]}} . Upon substituting this in the entropy inequality, we finally arrive at the inequality that must be satisfied by {{}_e mathbf{?}^{[0]}} , {{}_d mathbf{?}^{[0]}} and {mathbf{q}} . Derivations of the constitutive theory for {{}_e mathbf{?}^{[0]}} follow directly from {{}_e mathbf{?}^{(0)}} , equilibrium Cauchy stress tensor, and the constitutive theory for {mathbf{\\varepsilon}_{[n]}} is derived using the theory of generators and invariants. Constitutive theories for the heat vector {mathbf{q}} of up to orders n that are consistent (in terms of the argument tensors) with the constitutive theories for {mathbf{\\varepsilon}_{[n]}} are also derived. Many simplified forms of the rate theories of orders n are presented. Material coefficients are derived by considering Taylor series expansions of the coefficients in the linear combinations representing {mathbf{\\varepsilon}_{[n]}} and {mathbf{q}} using the combined generators of the argument tensors about a known configuration {\\underline{?}} in the combined invariants of the argument tensors and temperature. It is shown that the rate constitutive theories of order one (n = 1) when further simplified results in constitutive theories that resemble currently used theories but are in fact different. The solid materials characterized by these theories h

Surana, K. S.; Reddy, J. N.; Nunez, D.

2014-06-01

393

Adenoviral expression of a transforming growth factor-?1 antisense mRNA is effective in preventing liver fibrosis in bile-duct ligated rats  

Microsoft Academic Search

BACKGROUND: Transforming growth factor-? (TGF-?) is a key mediator in establishing liver fibrosis. Therefore, TGF-? as a causative agent may serve as a primary target for antifibrotic gene therapy approaches. We have previously shown that the adenoviral delivery of a transgene constitutively expressing a TGF-?1 antisense mRNA blocks TGF-? synthesis in culture-activated hepatic stellate cells and effectively abolishes ongoing fibrogenesis

Monica Arias; Sibille Sauer-Lehnen; Jens Treptau; Nora Janoschek; Ingo Theuerkauf; Reinhard Buettner; Axel M Gressner; Ralf Weiskirchen

2003-01-01

394

Development of operational models of receptor activation including constitutive receptor activity and their use to determine the efficacy of the chemokine CCL17 at the CC chemokine receptor CCR4  

PubMed Central

BACKGROUND AND PURPOSE The operational model provides a key conceptual framework for the analysis of pharmacological data. However, this model does not include constitutive receptor activity, a frequent phenomenon in modern pharmacology, particularly in recombinant systems. Here, we developed extensions of the operational model which include constitutive activity and applied them to effects of agonists at the chemokine receptor CCR4. EXPERIMENTAL APPROACH The effects of agonists of CCR4 on [35S]GTP?S binding to recombinant cell membranes and on the filamentous (F-) actin content of human CD4+ CCR4+ T cells were determined. The basal [35S]GTP?S binding was changed by varying the GDP concentration whilst the basal F-actin contents of the higher expressing T cell populations were elevated, suggesting constitutive activity of CCR4. Both sets of data were analysed using the mathematical models. RESULTS The affinity of CCL17 (also known as TARC) derived from analysis of the T cell data (pKa= 9.61 ± 0.17) was consistent with radioligand binding experiments (9.50 ± 0.11) while that from the [35S]GTP?S binding experiments was lower (8.27 ± 0.09). Its intrinsic efficacy differed between the two systems (110 in T cells vs. 11). CONCLUSIONS AND IMPLICATIONS The presence of constitutive receptor activity allows the absolute intrinsic efficacy of agonists to be determined without a contribution from the signal transduction system. Intrinsic efficacy estimated in this way is consistent with Furchgott's definition of this property. CCL17 may have a higher intrinsic efficacy at CCR4 in human T cells than that expressed recombinantly in CHO cells. PMID:22335621

Slack, RJ; Hall, DA

2012-01-01

395

Loi constitutive chimioplastique pour le beton expose aux hautes temperatures  

NASA Astrophysics Data System (ADS)

Concrete is the most widely used construction material in the world. Even though it has been used for several centuries, its behavior to high temperature remains to be understood. In the light of recent extreme events, including accidents, and arson, special attention has been focused on the performance of concrete in the fire safety assessment of buildings and tunnels. Fire represents one of the most severe conditions encountered during the life-time of a structure. Concrete exposed to high temperature can significantly jeopardize the structural integrity and load bearing capacity of the structure. Spalling of concrete remains one of the main issues to be addressed in the case of fire in buildings and tunnels. Successful modeling of this phenomenon depends not only on the accurate prediction of the temperature distribution through structural concrete but also on its mechanical response to the heating and boundaries restrains conditions and the migration of moisture and associated pore pressures. Therefore, it is necessary to develop a reliable formulation of concrete with all required information to understand its behavior during and after exposure to elevated temperature. It is also necessary to properly assess the effects of thermal degradation in order to develop predictive tools and validate design codes. Many structural problems can be adequately worthy by an elastoplastic model. The ultimate goal of this study is the development of a new constitutive model under a chemoplastic framework. To do this, an experimental program is carried out. The purpose of this program is twofold. First, it is essential to calibrate the proposed constitutive law that will be developed, and, second, for defining an inverse a problem. Usually, uniaxial and triaxial tests, conducted with confining pressure varied between 1.3 and 24 MPa and a temperature up to 700°C, allow us to identify the constitutive law parameters. This law reproduces the reduced field strength due to 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.

Hammoud, Rabah

396

Constitutive Activation of Stat3 Signaling Confers Resistance to Apoptosis in Human U266 Myeloma Cells  

Microsoft Academic Search

Interleukin 6 (IL-6) is the major survival factor for myeloma tumor cells and induces signaling through the STAT proteins. We report that one STAT family member, Stat3, is constitutively activated in bone marrow mononuclear cells from patients with multiple myeloma and in the IL-6-dependent human myeloma cell line U266. Moreover, U266 cells are inherently resistant to Fas-mediated apoptosis and express

Robyn Catlett-Falcone; Terry H Landowski; Marc M Oshiro; James Turkson; Alexander Levitzki; Rocco Savino; Gennaro Ciliberto; Lynn Moscinski; Jose Luis Fernández-Luna; Gabriel Nuńez; William S Dalton; Richard Jove

1999-01-01

397

Construction of a Novel Constitutively Active Chimeric EGFR to Identify New Targets for Therapy1  

Microsoft Academic Search

Tumor cells and tumor-associated endothelial cells express activated epidermal growth factor receptor (EGFR) due to production of EGF-related ligands in the tumor microenvironment. To investigate the effect of perpetual EGFR activation on endothelial cells, we developed a novel method to generate constitutively activeEGFR.Wefusedtheentireintracellulardomainof the EGFR to the N-terminus of the CD3Z component of theT-cellreceptorsignalingcomplex.Expressionofthe chimeric receptor CD3-EGFR in EGFR-deficient human

Hua Cheng; Robert R. Langley; Qiuyu Wu; Wenjuan Wu; Jie Feng; Rachel Tsan; Dominic Fan; Isaiah J. Fidler

398

Molecular pharmacological phenotyping of EBI2. An orphan seven-transmembrane receptor with constitutive activity.  

PubMed

Epstein-Barr virus (EBV)-induced receptor 2 (EBI2) is an orphan seven-transmembrane (7TM) receptor originally identified as the most up-regulated gene (>200-fold) in EBV-infected cells. Here we show that EBI2 signals with constitutive activity through Galpha(i) as determined by a receptor-mediated inhibition of forskolin-induced cAMP production and an induction of the serum response element-driven transcriptional activity in a pertussis toxin-sensitive manner. Galpha(s) and Galpha(q) were not activated constitutively as determined by the lack of cAMP production, the lack of inositol phosphate turnover, and the lack of activities of the transcription factors: cAMP response element-binding protein and nuclear factor-kappaB. Immunohistochemistry and confocal microscopy of FLAG- and green fluorescent protein-tagged EBI2 revealed cell-surface expression. A putative N-terminal truncated version of EBI2, delta4-EBI2, showed similar expression and signaling through Galpha(i) as full-length EBI2. By using a 32P-labeled EBI2 probe we found a very high expression in lymphoid tissue (spleen and lymph node) and peripheral blood mononuclear cells and a high expression in lung tissue. Real-time PCR of EBV-infected cells showed high expression of EBI2 during latent and lytic infection, in contrast to the EBV-encoded 7TM receptor BILF1, which was induced during lytic infection. EBI2 clustered with the orphan GPR18 by alignment analysis as well as by close proximity in the chromosomal region 13q32.3. Based on the constitutive signaling and cellular expression pattern of EBI2, it is suggested that it may function in conjunction with BILF1 in the reprogramming of the cell during EBV infection. PMID:16540462

Rosenkilde, Mette M; Benned-Jensen, Tau; Andersen, Helene; Holst, Peter J; Kledal, Thomas N; Lüttichau, Hans R; Larsen, Jřrgen K; Christensen, Jan P; Schwartz, Thue W

2006-05-12

399

Isolation and characterization of Schistosoma mansoni constitutive androstane receptor.  

PubMed

Full length cDNA clones encoding a Schistosoma mansoni homologue of vertebrate CAR/PXR/VDR group nuclear receptor, termed SmCAR were isolated from screening a S. mansoni adult worm cDNA library. SmCAR is a 702 amino acid protein which retains a typical domain organization of nuclear receptor superfamily members. A homology search demonstrated that SmCAR exhibits the highest homology with mouse constitutive androstane receptor (CAR). Like its orthologues from invertebrates, SmCAR contains a P box sequence of ESCKA in the DNA binding domain. The P box is important in determining the DNA binding specificity for nuclear receptors. SmCAR mRNA is expressed in every stage of S. mansoni life cycle with an elevated expression level in egg and cercaria stages. Two forms (78 and 81 kDa) of SmCAR protein were detected in schistosome worm extract by Western blot analysis. SmCAR protein was demonstrated to be widely distributed in adult worms by immunolocalization studies, being found in the subtegument in both male and female worms and in the ovaries, vitellaria and eggs in female worms. In vitro DNA binding assays demonstrated that SmCAR binds to the hsp27 ecdysone response element (EcRE) as well as schistosome p14 gene upstream region. The AGTGCA half site is essential for binding of SmCAR to the p14 gene upstream region. Therefore, AGTGCA probably serves as a high affinity binding half site for ESCKA containing nuclear receptors. PMID:16603257

Hu, Rong; Wu, Wenjie; Niles, Edward G; Loverde, Philip T

2006-07-01

400

Constitutive Endocytosis and Degradation of the Pre-T Cell Receptor  

PubMed Central

The pre-T cell receptor (TCR) signals constitutively in the absence of putative ligands on thymic stroma and signal transduction correlates with translocation of the pre-TCR into glycolipid-enriched microdomains (rafts) in the plasma membrane. Here, we show that the pre-TCR is constitutively routed to lysosomes after reaching the cell surface. The cell-autonomous down-regulation of the pre-TCR requires activation of the src-like kinase p56lck, actin polymerization, and dynamin. Constitutive signaling and degradation represents a feature of the pre-TCR because the ??TCR expressed in the same cell line does not exhibit these features. This is also evident by the observation that the protein adaptor/ubiquitin ligase c-Cbl is phosphorylated and selectively translocated into rafts in pre-TCR– but not ??TCR-expressing cells. A role of c-Cbl–mediated ubiquitination in pre-TCR degradation is supported by the reduction of degradation through pharmacological inhibition of the proteasome and through a dominant-negative c-Cbl ubiquitin ligase as well as by increased pre-TCR surface expression on immature thymocytes in c-Cbl–deficient mice. The pre-TCR internalization contributes significantly to the low surface level of the receptor on developing T cells, and may in fact be a requirement for optimal pre-TCR function. PMID:12070286

Panigada, Maddalena; Porcellini, Simona; Barbier, Eliane; Hoeflinger, Sonja; Cazenave, Pierre-André; Gu, Hua; Band, Hamid; von Boehmer, Harald; Grassi, Fabio

2002-01-01

401

Constitutive endocytosis and degradation of the pre-T cell receptor.  

PubMed

The pre-T cell receptor (TCR) signals constitutively in the absence of putative ligands on thymic stroma and signal transduction correlates with translocation of the pre-TCR into glycolipid-enriched microdomains (rafts) in the plasma membrane. Here, we show that the pre-TCR is constitutively routed to lysosomes after reaching the cell surface. The cell-autonomous down-regulation of the pre-TCR requires activation of the src-like kinase p56(lck), actin polymerization, and dynamin. Constitutive signaling and degradation represents a feature of the pre-TCR because the gammadeltaTCR expressed in the same cell line does not exhibit these features. This is also evident by the observation that the protein adaptor/ubiquitin ligase c-Cbl is phosphorylated and selectively translocated into rafts in pre-TCR- but not gammadeltaTCR-expressing cells. A role of c-Cbl-mediated ubiquitination in pre-TCR degradation is supported by the reduction of degradation through pharmacological inhibition of the proteasome and through a dominant-negative c-Cbl ubiquitin ligase as well as by increased pre-TCR surface expression on immature thymocytes in c-Cbl-deficient mice. The pre-TCR internalization contributes significantly to the low surface level of the receptor on developing T cells, and may in fact be a requirement for optimal pre-TCR function. PMID:12070286

Panigada, Maddalena; Porcellini, Simona; Barbier, Eliane; Hoeflinger, Sonja; Cazenave, Pierre-André; Gu, Hua; Band, Hamid; von Boehmer, Harald; Grassi, Fabio

2002-06-17

402

Constitutive regulatory activity of an evolutionarily excluded riboswitch variant.  

PubMed

The exquisite specificity of the adenine-responsive riboswitch toward its cognate metabolite has been shown to arise from the formation of a Watson-Crick interaction between the adenine ligand and residue U65. A recent crystal structure of a U65C adenine aptamer variant has provided a rationale for the phylogenetic conservation observed at position 39 for purine aptamers. The G39-C65 variant adopts a compact ligand-free structure in which G39 is accommodated by the ligand binding site and is base-paired to the cytosine at position 65. Here, we demonstrate using a combination of biochemical and biophysical techniques that the G39-C65 base pair not only severely impairs ligand binding but also disrupts the functioning of the riboswitch in vivo by constitutively activating gene expression. Folding studies using single-molecule FRET revealed that the G39-C65 variant displays a low level of dynamic heterogeneity, a feature reminiscent of ligand-bound wild-type complexes. A restricted conformational freedom together with an ability to significantly fold in monovalent ions are exclusive to the G39-C65 variant. This work provides a mechanistic framework to rationalize the evolutionary exclusion of certain nucleotide combinations in favor of sequences that preserve ligand binding and gene regulation functionalities. PMID:21676871

Tremblay, Renaud; Lemay, Jean-François; Blouin, Simon; Mulhbacher, Jérôme; Bonneau, Éric; Legault, Pascale; Dupont, Paul; Penedo, J Carlos; Lafontaine, Daniel A

2011-08-01

403

Dual action of oestrogens on the mouse constitutive androstane receptor.  

PubMed Central

mCAR (mouse constitutive androstane receptor; NR1I3) controls the expression of cytochrome P450 as well as other enzymes involved in drug and steroid metabolism. The high basal activity of mCAR can be modulated by inhibitory steroids related to androstenol and by activating xenobiotic chemicals such as 1,4-bis-[2-(3,5-dichloropyridyloxy)]benzene and chlorpromazine. The ability of oestrogens and some other xenobiotics to activate mCAR is not clear. In the present study, co-transfection assays in HEK-293 cells indicated that oestrogens varied in their efficacy to activate mCAR, depending on variation at the steroid D-ring and position of hydroxy groups. In general, oestrogens were weaker activators of mCAR than 1,4-bis-[2-(3,5-dichloropyridyloxy)]benzene and chlorpromazine. Also, the induction of CYP2B10 mRNA by oestrogens was less pronounced in mouse primary hepatocytes. Yeast two-hybrid assays indicated that, unlike androstenol and the established activators, oestrogens attracted both nuclear receptor co-repressors and co-activators to the mCAR ligand-binding domain, thus limiting the extent of mCAR activation. This novel dual action is not limited to oestrogens, but is shared by some xenobiotic CYP2B inducers such as clotrimazole and methoxychlor. These findings offer an alternative explanation for the recently suggested nuclear activation step of mCAR. PMID:12948398

Mäkinen, Janne; Reinisalo, Mika; Niemi, Kaisa; Viitala, Pirkko; Jyrkkärinne, Johanna; Chung, Hinfan; Pelkonen, Olavi; Honkakoski, Paavo

2003-01-01

404

The human constitutive androstane receptor promotes the differentiation and maturation of hepatic-like cells  

PubMed Central

Expression of the constitutive androstane receptor (CAR, NR1I3) is enriched in the mature mammalian liver and increasingly recognized for its prominent role in regulating a myriad of processes including biotransformation, chemical transport, energy metabolism and lipid homeostasis. Previously, we demonstrated that CAR levels were markedly enhanced during the differentiation of hepatic-like cells derived from hESCs, prompting the hypothesis that CAR contributes a key functional role in directing human hepatogenesis. Here we demonstrate that over-expression of CAR in human embryonic stem cells (ESCs), transduced by a lentiviral vector, accelerates the maturation of hepatic-like cells, with CAR over-expressing cells exhibiting a 2.5-fold increase in albumin secretion by day 20 in culture differentiation, and significantly enhanced levels of mRNA expression of several liver-selective markers, including hepatic transcription factors, plasma proteins, biotransformation enzymes, and metabolic enzymes. CAR over-expressing cells also exhibited enhanced CITCO-inducible CYP3A7 enzymatic activity. Knockdown of CAR via siRNA attenuated the differentiation-dependent expression programs. In contrast, expression levels of the pregnane X receptor (PXR), a nuclear receptor most similar to CAR in primary sequence, were negligible in human fetal liver tissues or in the differentiating hESCs, and stable over-expression of PXR in hepatic-induced hESCs failed to enhance expression of hepatic phenotype markers. Together, these results define a novel role for human CAR in hepatic lineage commitment. PMID:24144921

Chen, Fengming; Zamule, Stephanie M.; Coslo, Denise M.; Chen, Tao; Omiecinski, Curtis J.

2013-01-01

405

MLH1 constitutional and somatic methylation in patients with MLH1 negative tumors fulfilling the revised Bethesda criteria.  

PubMed

Lynch syndrome (LS) is a tumor predisposing condition caused by constitutional defects in genes coding for components of the mismatch repair (MMR) apparatus. While hypermethylation of the promoter of the MMR gene MLH1 occurs in about 15% of colorectal cancer samples, it has also been observed as a constitutional alteration, in the absence of DNA sequence mutations, in a small number of LS patients. In order to obtain further insights on the phenotypic characteristics of MLH1 epimutation carriers, we investigated the somatic and constitutional MLH1 methylation status of 14 unrelated subjects with a suspicion of LS who were negative for MMR gene constitutional mutations and whose tumors did not express the MLH1 protein. A novel case of constitutional MLH1 epimutation was identified. This patient was affected with multiple primary tumors, including breast cancer, diagnosed starting from the age of 55 y. Investigation of her offspring by allele specific expression revealed that the epimutation was not stable across generations. We also found MLH1 hypermethylation in cancer samples from 4 additional patients who did not have evidence of constitutional defects. These patients had some characteristics of LS, namely early age at onset and/or positive family history, raising the possibility of genetic influences in the establishment of somatic MLH1 methylation. PMID:25437057

Crucianelli, Francesca; Tricarico, Rossella; Turchetti, Daniela; Gorelli, Greta; Gensini, Francesca; Sestini, Roberta; Giunti, Laura; Pedroni, Monica; Ponz de Leon, Maurizio; Civitelli, Serenella; Genuardi, Maurizio

2014-10-01

406

Constitutive activation of AMPK ?1 in vascular endothelium promotes high-fat diet-induced fatty liver injury: role of COX-2 induction  

PubMed Central

Background and Purpose:?AMP-activated protein kinase (AMPK), an important regulator of energy metabolism, comprises three (?, ? and ?) subunits, each with a unique tissue distribution. As AMPK has a wide range of protein and gene targets, defining its role has been difficult. Here, we have studied a transgenic mouse model overexpressing the constitutively active ?1 subunit of AMPK in endothelial cells (EC-AMPK) to elucidate its role in energy homeostasis. Experimental Approach:?Wild-type and EC-AMPK mice were fed with a high fat diet for 16 weeks. Drugs (or vehicles) were given daily by oral gavage. Body weight, fat mass composition, glucose and lipid levels were monitored regularly. Tissues including aortae and liver were collected for quantitative RT-PCR, Western blotting, elisa, histological and biochemical evaluations. Key Results:?Compared with wild-type animals, high fat diet caused more severe metabolic defects in EC-AMPK mice, which exhibited increased body weight and fat mass, elevated blood pressure, augmented glucose and lipid levels, impaired glucose tolerance, hepatomegaly and steatohepatitis. Constitutive activation of AMPK ?1 in endothelial cells induced COX-2 expression and arterial inflammation. Genes involved in lipid metabolism were down-regulated in aortae and livers of EC-AMPK mice. Chronic treatment with selective COX-2 inhibitors, celecoxib or nimesulide, significantly ameliorated arterial inflammation, steatohepatitis and hyperlipidaemia in EC-AMPK mice, without altering their blood pressure or clotting. Conclusions and Implications:?Constitutive activation of endothelial AMPK ?1 promotes vascular inflammation and the development of obesity-induced fatty livers largely via induction of COX-2. PMID:24372551

Liang, Yan; Huang, Bosheng; Song, Erfei; Bai, Bo; Wang, Yu

2014-01-01

407

Integration of constitutive equations with a new explicit scheme  

Microsoft Academic Search

Precise integration of a constitutive law is required whenever a boundary value problem is to be solved. In this paper a new\\u000a explicit scheme for the integration of elasto-plastic constitutive models, that consist of algebraic yield condition and differential\\u000a evolution equations, is presented. The scheme was primarily developed for the integration of constitutive models in boundary\\u000a value problems which are

Miroslav Halilovi?; Marko Vrh; Boris Štok

2010-01-01

408

Constitutive activation of RANK disrupts mammary cell fate leading to tumorigenesis.  

PubMed

Receptor Activator of NF-kappa B (RANK) pathway controls mammary gland development in mice but its role in mammary stem cell fate remains undefined. We show that constitutive RANK signaling expands luminal and basal mammary compartments including mammary stem and luminal progenitor cell pools and interferes with the generation of CD61+ and Sca1+ luminal cells and Elf5 expression. Impaired mammary cell commitment upon RANK overexpression leads to the accumulation of progenitors including K14+K8+ bipotent cells and the formation of heterogeneous tumors containing hyperplastic basal, luminal, and progenitor cells. RANK expression increases in wild-type mammary epithelia with age and parity, and spontaneous preneoplastic lesions express RANK and accumulate K14+K8+ cells. In human breast tumors, high RANK expression levels are also associated with altered mammary differentiation. These results suggest that increased RANK signaling interferes with mammary cell commitment, contributing to breast carcinogenesis. PMID:23766243

Pellegrini, Pasquale; Pasquale, Pellegrini; Cordero, Alex; Alex, Cordero; Gallego, Marta Ines; Marta Ines, Gallego; Dougall, William C; William, Dougall C; Purificación, Muńoz; Pujana, Miguel Angel; Angel, Pujana Miguel; Gonzalez-Suarez, Eva; Eva, Gonzalez-Suarez

2013-09-01

409

Global Transcriptome Analysis of Constitutive Resistance to the White Pine Weevil in Spruce  

PubMed Central

Constitutive defense mechanisms are critical to the understanding of defense mechanisms in conifers because they constitute the first barrier to attacks by insect pests. In interior spruce, trees that are putatively resistant and susceptible to attacks by white pine weevil (Pissodes strobi) typically exhibit constitutive differences in traits such as resin duct size and number, bark thickness, and terpene content. To improve our knowledge of their genetic basis, we compared globally the constitutive expression levels of 17,825 genes between 20 putatively resistant and 20 putatively susceptible interior spruce trees from the British Columbia tree improvement program. We identified 54 upregulated and 137 downregulated genes in resistant phenotypes, relative to susceptible phenotypes, with a maximum fold change of 2.24 and 3.91, respectively. We found a puzzling increase of resistance by downregulated genes, as one would think that “procuring armaments” is the best defense. Also, although terpenes and phenolic compounds play an important role in conifer defense, we found few of these genes to be differentially expressed. We found 15 putative small heat-shock proteins (sHSP) and several other stress-related proteins to be downregulated in resistant trees. Downregulated putative sHSP belong to several sHSP classes and represented 58% of all tested putative sHSP. These proteins are well known to be involved in plant response to various kinds of abiotic stress; however, their role in constitutive resistance is not yet understood. The lack of correspondence between transcriptome profile clusters and phenotype classifications suggests that weevil resistance in spruce is a complex trait. PMID:21852250

Verne, Sébastien; Jaquish, Barry; White, Rick; Ritland, Kermit

2011-01-01

410

Nonlinear Constitutive Relations for High Temperature Application, 1984  

NASA Technical Reports Server (NTRS)

Nonlinear constitutive relations for high temperature applications were discussed. The state of the art in nonlinear constitutive modeling of high temperature materials was reviewed and the need for future research and development efforts in this area was identified. Considerable research efforts are urgently needed in the development of nonlinear constitutive relations for high temperature applications prompted by recent advances in high temperature materials technology and new demands on material and component performance. Topics discussed include: constitutive modeling, numerical methods, material testing, and structural applications.

1985-01-01

411

Constitutive equations for discrete electromagnetic problems over polyhedral grids  

SciTech Connect

In this paper a novel approach is proposed for constructing discrete counterparts of constitutive equations over polyhedral grids which ensure both consistency and stability of the algebraic equations discretizing an electromagnetic field problem. The idea is to construct discrete constitutive equations preserving the thermodynamic relations for constitutive equations. In this way, consistency and stability of the discrete equations are ensured. At the base, a purely geometric condition between the primal and the dual grids has to be satisfied for a given primal polyhedral grid, by properly choosing the dual grid. Numerical experiments demonstrate that the proposed discrete constitutive equations lead to accurate approximations of the electromagnetic field.

Codecasa, Lorenzo [Politecnico di Milano, Piazza Leonardo da Vinci 32, I-20133 Milan (Italy)]. E-mail: codecasa@elet.polimi.it; Trevisan, Francesco [Universita di Udine, Via delle Scienze 208, I-33100 Udine (Italy)]. E-mail: trevisan@uniud.it

2007-08-10

412

Beam induced backgrounds: CDF experience  

SciTech Connect

We summarize the experiences of the Collider Detector at Fermilab (CDF) experiment in the presence of backgrounds originating from the counter circulating beams in the Fermilab Tevatron. These backgrounds are measured and their sources identified. Finally, we outline the strategies employed to reduce the effects of these backgrounds on the experiment.

Tesarek, R.J.; /Fermilab

2008-05-01

413

Nonlocal constitutive relation for steady granular flow.  

PubMed

Extending recent modeling efforts for emulsions, we propose a nonlocal fluidity relation for flowing granular materials, capturing several known finite-size effects observed in steady flow. We express the local Bagnold-type granular flow law in terms of a fluidity ratio and then extend it with a particular Laplacian term that is scaled by the grain size. The resulting model is calibrated against a sequence of existing discrete element method data sets for two-dimensional annular shear, where it is shown that the model correctly describes the divergence from a local rheology due to the grain size as well as the rate-independence phenomenon commonly observed in slowly flowing zones. The same law is then applied in two additional inhomogeneous flow geometries, and the predicted velocity profiles are compared against corresponding discrete element method simulations utilizing the same grain composition as before, yielding favorable agreement in each case. PMID:22680912

Kamrin, Ken; Koval, Georg

2012-04-27

414

Modeling the constitutive response of bimodal metals  

NASA Astrophysics Data System (ADS)

The mechanical response of metals with a bimodal grain-size distribution is modeled using the secant Mori-Tanaka (M-T) mean-field approach. The actual microstructure of bimodal metals involves a grain size distribution in the ultrafine and coarse regimes; the model approximates this in terms of two phases with distinct grain sizes and with specific volume fractions. The model is applied to two bimodal materials: the Al-5083 alloys of Lavernia et al. and the Cu of Wang et al. In both the materials, the predictions agree well with the experiments. In the bimodal Al alloy, the effect of extrusion on the anisotropy in yield strength and flow behavior is also addressed. Finally, based on the model predictions, an empirical expression of the Voce form is proposed to describe the overall flow behavior of both bimodal metals.

Joshi, S. P.; Ramesh, K. T.; Han, B. Q.; Lavernia, E. J.

2006-08-01

415

Sucrose: a constitutive elicitor of phytoalexin synthesis.  

PubMed

Extracts of seeds and leaves of the tropical legume Cajanus cajan (L.) Millsp. (the pigeon pea) elicited the accumulation of three phytoalexins when applied as droplets to superficially wounded leaves of the plant. The active component was purified and identified as sucrose. Phytoalexin accumulation was proportional to the logarithm of the concentration of sucrose applied, with maxima ranging from 338 to 455 micrograms per gram (fresh weight) of leaf tissue. The sucrose concentrations required to elicit half these amounts ranged from 20 to 35 micrograms per milliliter, but other sugars had little effect even at 1000 micrograms per milliliter. The elicitor activity of sucrose was abolished by actinomycin D, puromycin, and cycloheximide at a concentration of 10 micrograms per milliliter or greater, suggesting that gene derepression is required for expression of the phytoalexin response. PMID:17730656

Cooksey, C J; Garratt, P J; Dahiya, J S; Strange, R N

1983-06-24

416

Indiana v. Jamie L. Curtis: "The Case of the Questionable Book Bag Search." A Constitution-based Scripted Trial.  

ERIC Educational Resources Information Center

This document presents "Indiana v. Jamie L. Curtis," a fictional court case stemming from the search by school officials of a student's bookbag. The case is based upon the actual U.S. Supreme Court case of New Jersey v. T.L.O., which involved similar facts. The document includes background materials on the Fourth Amendment to the U.S. Constitution

Leming, Robert S.

417

Background issues for defensive interceptors  

SciTech Connect

Mean nuclear backgrounds are large, but are arguably amenable to frame-to-frame subtraction. Striated backgrounds on the sensors for defensive interceptors could, however, cause clutter leak-through, which could make detection and track difficult. Nominal motions and backgrounds give signal to clutter ratios too low to be useful. Clutter leakage due to line-of-sight drift can be reduced by stabilizing the line of sight around the background clutter itself. Current interceptors have detector arrays large enough for operation independent of nuclear backgrounds in their fields of view. 6 refs., 2 figs.

Canavan, G.H.

1991-03-01

418

Mining microarray expression data by literature profiling  

PubMed Central

Background The rapidly expanding fields of genomics and proteomics have prompted the development of computational methods for managing, analyzing and visualizing expression data derived from microarray screening. Nevertheless, the lack of efficient techniques for assessing the biological implications of gene-expression data remains an important obstacle in exploiting this information. Results To address this need, we have developed a mining technique based on the analysis of literature profiles generated by extracting the frequencies of certain terms from thousands of abstracts stored in the Medline literature database. Terms are then filtered on the basis of both repetitive occurrence and co-occurrence among multiple gene entries. Finally, clustering analysis is performed on the retained frequency values, shaping a coherent picture of the functional relationship among large and heterogeneous lists of genes. Such data treatment also provides information on the nature and pertinence of the associations that were formed. Conclusions The analysis of patterns of term occurrence in abstracts constitutes a means of exploring the biological significance of large and heterogeneous lists of genes. This approach should contribute to optimizing the exploitation of microarray technologies by providing investigators with an interface between complex expression data and large literature resources. PMID:12372143

Chaussabel, Damien; Sher, Alan

2002-01-01

419

The Epistemological Background of our Strategies  

NASA Astrophysics Data System (ADS)

The major objectives of the long-term strategy in promoting a scientific discipline into the education sphere may be the following : • A permanent presence of the discipline inside the general education • A good harmonisation with other disciplines (at least the scientific ones) • A maximal efficiency of its presence as a required discipline Before any practical approaches (social, administrative, or any other), it is required (necessary) that the promoters of any discipline formulate and underline its cognitive and formative contribution. This can be done only by starting with the epistemological statute of the discipline and its pedagogical implications. Defining the epistemological statute, as well as the requirements of an optimal communication among the options concerning the orientation (re-orientation) of educational strategies, brings forth the necessity of analysing some fundamental concepts, with epistemological characters. Even if it seems to be known by everyone, usually the concepts are not taken into account with their real importance. The fundamental concept we refer to herein constitute a real "conceptual network" who, "volens-nolens", is the epistemological background for any strategy of education relative to science. Being conscious of the importance of the epistemological background will permit the modification of the beginning and the references used in defining the strategy of education in (through) astronomy.

Birlan, M.; Vass, G.; Teleanu, C.

2006-08-01

420

Constitutive and ligand-induced EGFR signalling triggers distinct and mutually exclusive downstream signalling networks.  

PubMed

Epidermal growth factor receptor (EGFR) overexpression plays an important oncogenic role in cancer. Regular EGFR protein levels are increased in cancer cells and the receptor then becomes constitutively active. However, downstream signals generated by constitutively activated EGFR are unknown. Here we report that the overexpressed EGFR oscillates between two distinct and mutually exclusive modes of signalling. Constitutive or non-canonical EGFR signalling activates the transcription factor IRF3 leading to expression of IFI27, IFIT1 and TRAIL. Ligand-mediated activation of EGFR switches off IRF3-dependent transcription, activates canonical extracellular signal-regulated kinase (ERK) and Akt signals, and confers sensitivity to chemotherapy and virus-induced cell death. Mechanistically, the distinct downstream signals result from a switch of EGFR-associated proteins. EGFR constitutively complexes with IRF3 and TBK1 leading to TBK1 and IRF3 phosphorylation. Addition of epidermal growth factor dissociates TBK1, IRF3 and EGFR leading to a loss of IRF3 activity, Shc-EGFR association and ERK activation. Finally, we provide evidence for non-canonical EGFR signalling in glioblastoma. PMID:25503978

Chakraborty, Sharmistha; Li, Li; Puliyappadamba, Vineshkumar Thidil; Guo, Gao; Hatanpaa, Kimmo J; Mickey, Bruce; Souza, Rhonda F; Vo, Peggy; Herz, Joachim; Chen, Mei-Ru; Boothman, David A; Pandita, Tej K; Wang, David H; Sen, Ganes C; Habib, Amyn A

2014-01-01