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  1. Stably Expressed Genes Involved in Basic Cellular Functions.

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    Kejian Wang

    Full Text Available Stably Expressed Genes (SEGs whose expression varies within a narrow range may be involved in core cellular processes necessary for basic functions. To identify such genes, we re-analyzed existing RNA-Seq gene expression profiles across 11 organs at 4 developmental stages (from immature to old age in both sexes of F344 rats (n = 4/group; 320 samples. Expression changes (calculated as the maximum expression / minimum expression for each gene of >19000 genes across organs, ages, and sexes ranged from 2.35 to >109-fold, with a median of 165-fold. The expression of 278 SEGs was found to vary ≤4-fold and these genes were significantly involved in protein catabolism (proteasome and ubiquitination, RNA transport, protein processing, and the spliceosome. Such stability of expression was further validated in human samples where the expression variability of the homologous human SEGs was significantly lower than that of other genes in the human genome. It was also found that the homologous human SEGs were generally less subject to non-synonymous mutation than other genes, as would be expected of stably expressed genes. We also found that knockout of SEG homologs in mouse models was more likely to cause complete preweaning lethality than non-SEG homologs, corroborating the fundamental roles played by SEGs in biological development. Such stably expressed genes and pathways across life-stages suggest that tight control of these processes is important in basic cellular functions and that perturbation by endogenous (e.g., genetics or exogenous agents (e.g., drugs, environmental factors may cause serious adverse effects.

  2. Antigenic structures stably expressed by recombinant TGEV-derived vectors.

    Science.gov (United States)

    Becares, Martina; Sanchez, Carlos M; Sola, Isabel; Enjuanes, Luis; Zuñiga, Sonia

    2014-09-01

    Coronaviruses (CoVs) are positive-stranded RNA viruses with potential as immunization vectors, expressing high levels of heterologous genes and eliciting both secretory and systemic immune responses. Nevertheless, its high recombination rate may result in the loss of the full-length foreign gene, limiting their use as vectors. Transmissible gastroenteritis virus (TGEV) was engineered to express porcine reproductive and respiratory syndrome virus (PRRSV) small protein domains, as a strategy to improve heterologous gene stability. After serial passage in tissue cultures, stable expression of small PRRSV protein antigenic domains was achieved. Therefore, size reduction of the heterologous genes inserted in CoV-derived vectors led to the stable expression of antigenic domains. Immunization of piglets with these TGEV vectors led to partial protection against a challenge with a virulent PRRSV strain, as immunized animals showed reduced clinical signs and lung damage. Further improvement of TGEV-derived vectors will require the engineering of vectors with decreased recombination rate. Copyright © 2014 Elsevier Inc. All rights reserved.

  3. Characterization of new cell line stably expressing CHI3L1 oncogene

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    Chekhonin V. P.

    2011-06-01

    Full Text Available Aim. To characterize the immortalized 293 cell line after stable transfection with human oncogene (CHI3L1. Methods. 293 cells, stably transfected with pcDNA3.1_CHI3L1, and 293 cells, stably transfected with pcDNA3.1 as a negative control, were used throughout all experiments. The clones of CHI3L1-expressing 293 cells and 293 cells, transfected with pcDNA3.1, were analyzed by immunofluorescence and confocal microscopy. Cell proliferation was measured using MTT assay; analyses of ERK1/2 and AKT activation and their cellular localization were performed with anti-phospho-ERK and anti-phospho-AKT antibodies. Specific activation of MAP and PI3 kinases was measured by densitometric analysis of Western-blot signals. Results. The obtained results show quite modest ability of CHI3L1 to stimulate cell growth and reflect rather an improved cellular plating efficiency of the 293 cells stably transfected with pcDNA3.1_CHI3L1 as compared to the 293 cells transfected with an «empty» vector. ERK1/2 and AKT are activated in the 293_CHI3L1 cells. In these cells phosphorylated ERK1/2 were localized in both cell cytoplasm and nuclei while AKT only in cytoplasm. The 293_CHI3L1 cells differed from the 293 cells, transfected with an «empty» vector, in their size and ability to adhere to the culture plates. Conclusions. The overexpression of CHI3L1 is likely to have an important role in tumorigenesis via a mechanism which involves activation of PI3K and ERK1/2 pathways. The tumors which can be induced by orthotopic implantation of the transformed human cells with overexpressed human oncogene CHI3L1 into the rat brain can be used as a target for anticancer drug development.

  4. Identifying stably expressed housekeeping genes in the endometrium of fertile women, women with recurrent implantation failure and recurrent miscarriages

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    Stocker, Linden; Cagampang, Felino; Cheong, Ying

    2017-01-01

    Housekeeping genes (HKG) are presumed to be constitutively expressed throughout tissue types but recent studies have shown they vary with pathophysiology. Often, validation of appropriate HKG is not made. There is no consensus on which HKGs are most stably expressed in endometrial tissue so this study aimed to identify the most stable HKG in the endometrium of women with recurrent implantation failure (RIF) and recurrent miscarriages (RM). Inclusion criteria were women between 25-45 years (...

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

    DEFF Research Database (Denmark)

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

    2013-01-01

    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...

  6. Stably transfected human cell lines as fluorescent screening assay for nuclear factor KB activation dependent gene expression

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    Hellweg, Christine E.; Baumstark-Khan, Christa; Horneck, Gerda

    2004-06-01

    Activation of the Nuclear Factor kappaB (NF-kappaB) pathway as a possible antiapoptotic route represents one important cellular stress response. For identifying conditions which are capable to modify this pathway, a screening assay for detection of NF-kappaB-dependent gene activation using the reporter proteins Enhanced Green Fluorescent Protein (EGFP) and its destabilized variant (d2EGFP) has been developed. Human Embryonic Kidney (HEK/293) cells were stably transfected with a vector carrying EGFP or d2EGFP under control of a synthetic promoter containing four copies of the NF-kappaB response element. Treatment with tumor necrosis factor alpha (TNF-alpha) gave rise to substantial EGFP / d2EGFP expression in up to 90 % of the cells and was therefore used to screen different stably transfected clones for induction of NF-kappaB dependent gene expression. The time course of d2EGFP expression after treatment with TNF-alpha or phorbol ester was measured using flow cytometry. Cellular response to TNF-alpha was faster than to phorbol ester. Treatment of cells with TNF-alpha and DMSO revealed antagonistic interactions of these substances in the activation NF-kappaB dependent gene expression. The detection of d2EGFP expression required FACS analysis or fluorescence microscopy, while EGFP could also be measured in the microplate reader, rendering the assay useful for high-throughput screening.

  7. The antagonistic effect of antipsychotic drugs on a HEK293 cell line stably expressing human alpha(1A1)-adrenoceptors

    DEFF Research Database (Denmark)

    Nourian, Zahra; Mulvany, Michael J; Nielsen, Karsten Bork

    2008-01-01

    Antipsychotic drugs often cause orthostatic hypotension, probably through antagonist action on resistance vessel alpha(1A)-adrenoceptors. Here we have tested this possibility directly using cells transfected with a relevant human alpha(1A)-adrenoceptor splice variant. To determine a splice variant...... a cell line stably expressing a functional form of this splice variant. The expression of recombinant alpha(1A1)-adrenoceptor subtype was confirmed by Western immunoblot analysis, and its functionality demonstrated using a Fura-2 assay by a rise in intracellular calcium concentration ([Ca(2+)](i)) when...... human alpha(1A1)-adrenoceptors in competition binding studies confirmed much higher antagonist affinity of sertindole and risperidone than haloperidol for these receptors. In summary, it can be concluded that there is an approximately 10-fold higher adrenoceptor affinity of risperidone and sertindole...

  8. [Yeast (Saccharomyces cerevisiae) secretory expression vector maintained stably in Pro3+ transformants in rich medium].

    Science.gov (United States)

    Xie, H Y; Tang, Y; Jiang, W D; He, H Y; Liu, M; Kuang, D R

    2000-01-01

    A yeast (Saccharomyces cerevisiae) secretory expression vector containing PRO3 gene was constructed (pCBy310). beta HCG(Human choriogonadotropin beta subunit)-cDNA was inserted into pCBy310 to form a recombinant plasmid pCBy314. Since yeast proline auxotroph will not survive in rich medium (YPD), YPD could be used as a selection pressure, and pCBy314 could be maintained mitotically stable in transformants of yeast Pro3- auxotroph (MB299-7A) in rich medium. At an improved, yet not optimized cultural condition, the expression of beta HCG in culture medium was 650 micrograms/L. Our results showed not only that YPD could be used as a selection medium, but also that yeast grew better in YPD so as to increase the gene expression product, and that the component of YPD was simple and cheap. Our data suggested that PRO genes might be used widely in constructing vectors to clone and express foreign genes in yeast so that rich medium can be used as a selection pressure for direct selection.

  9. Cholesterol Down-Regulates BK Channels Stably Expressed in HEK 293 Cells

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    Deng, Xiu-Ling; Sun, Hai-Ying; Li, Gui-Rong

    2013-01-01

    Cholesterol is one of the major lipid components of the plasma membrane in mammalian cells and is involved in the regulation of a number of ion channels. The present study investigates how large conductance Ca2+-activated K+ (BK) channels are regulated by membrane cholesterol in BK-HEK 293 cells expressing both the α-subunit hKCa1.1 and the auxiliary β1-subunit or in hKCa1.1-HEK 293 cells expressing only the α-subunit hKCa1.1 using approaches of electrophysiology, molecular biology, and immunocytochemistry. Membrane cholesterol was depleted in these cells with methyl-β-cyclodextrin (MβCD), and enriched with cholesterol-saturated MβCD (MβCD-cholesterol) or low-density lipoprotein (LDL). We found that BK current density was decreased by cholesterol enrichment in BK-HEK 293 cells, with a reduced expression of KCa1.1 protein, but not the β1-subunit protein. This effect was fully countered by the proteasome inhibitor lactacystin or the lysosome function inhibitor bafilomycin A1. Interestingly, in hKCa1.1-HEK 293 cells, the current density was not affected by cholesterol enrichment, but directly decreased by MβCD, suggesting that the down-regulation of BK channels by cholesterol depends on the auxiliary β1-subunit. The reduced KCa1.1 channel protein expression was also observed in cultured human coronary artery smooth muscle cells with cholesterol enrichment using MβCD-cholesterol or LDL. These results demonstrate the novel information that cholesterol down-regulates BK channels by reducing KCa1.1 protein expression via increasing the channel protein degradation, and the effect is dependent on the auxiliary β1-subunit. PMID:24260325

  10. Stably expressed housekeeping genes across developmental stages in the two-spotted spider mite, Tetranychus urticae.

    Science.gov (United States)

    Yang, Chunxiao; Pan, Huipeng; Liu, Yong; Zhou, Xuguo

    2015-01-01

    Quantitative real-time PCR (qRT-PCR) is a reliable and reproducible technique for measuring mRNA expression. To facilitate gene expression studies and obtain more accurate qRT-PCR analysis, normalization relative to stable housekeeping genes is mandatory. In this study, ten housekeeping genes, including beta-actin (Actin) , elongation factor 1 α (EF1A) , glyceralde hyde-3-phosphate dehydrogenase (GAPDH) , ribosomal protein L13 (RPL13) , ribosomal protein 49 (RP49) , α-tubulin (Tubulin) , vacuolar-type H+-ATPase (v-ATPase) , succinate dehydrogenase subunit A (SDHA) , 28S ribosomal RNA (28S) , and 18S ribosomal RNA (18S) from the two-spotted spider mite, Tetranychus urticae, were selected as the candidate reference genes. Four algorithms, geNorm, Normfinder, BestKeeper, and the ΔCt method, were used to evaluate the performance of these candidates as endogenous controls across different developmental stages. In addition, RefFinder, which integrates the above-mentioned software tools, provided the overall ranking of the stability/suitability of these candidate reference genes. Among them, PRL13 and v-ATPase were the two most stable housekeeping genes across different developmental stages. This work is the first step toward establishing a standardized qRT-PCR analysis in T. urticae following the MIQE guideline. With the recent release of the T. urticae genome, results from this study provide a critical piece for the subsequent genomics and functional genomics research in this emerging model system.

  11. Biophysical and Pharmacological Characterization of Nav1.9 Voltage Dependent Sodium Channels Stably Expressed in HEK-293 Cells.

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    Zhixin Lin

    Full Text Available The voltage dependent sodium channel Nav1.9, is expressed preferentially in peripheral sensory neurons and has been linked to human genetic pain disorders, which makes it target of interest for the development of new pain therapeutics. However, characterization of Nav1.9 pharmacology has been limited due in part to the historical difficulty of functionally expressing recombinant channels. Here we report the successful generation and characterization of human, mouse and rat Nav1.9 stably expressed in human HEK-293 cells. These cells exhibit slowly activating and inactivating inward sodium channel currents that have characteristics of native Nav1.9. Optimal functional expression was achieved by coexpression of Nav1.9 with β1/β2 subunits. While recombinantly expressed Nav1.9 was found to be sensitive to sodium channel inhibitors TC-N 1752 and tetracaine, potency was up to 100-fold less than reported for other Nav channel subtypes despite evidence to support an interaction with the canonical local anesthetic (LA binding region on Domain 4 S6. Nav1.9 Domain 2 S6 pore domain contains a unique lysine residue (K799 which is predicted to be spatially near the local anesthetic interaction site. Mutation of this residue to the consensus asparagine (K799N resulted in an increase in potency for tetracaine, but a decrease for TC-N 1752, suggesting that this residue can influence interaction of inhibitors with the Nav1.9 pore. In summary, we have shown that stable functional expression of Nav1.9 in the widely used HEK-293 cells is possible, which opens up opportunities to better understand channel properties and may potentially aid identification of novel Nav1.9 based pharmacotherapies.

  12. Identifying stably expressed housekeeping genes in the endometrium of fertile women, women with recurrent implantation failure and recurrent miscarriages.

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    Stocker, Linden; Cagampang, Felino; Cheong, Ying

    2017-11-01

    Housekeeping genes (HKG) are presumed to be constitutively expressed throughout tissue types but recent studies have shown they vary with pathophysiology. Often, validation of appropriate HKG is not made. There is no consensus on which HKGs are most stably expressed in endometrial tissue so this study aimed to identify the most stable HKG in the endometrium of women with recurrent implantation failure (RIF) and recurrent miscarriages (RM). Inclusion criteria were women between 25-45 years (n = 45) suffering recurrent miscarriage (RM), recurrent implantation failure (RIF) or fertile controls. Endometrial biopsies were taken and total RNA extraction, cDNA synthesis and PCR was performed using 10 candidate HKG. The genes were arranged in terms of stability and normalisation was determined. Several HKGs not previously tested in endometrial samples were found to be more stable than those previously identified as the most stable. Of these, the 5 most stable HKG (in order of stability) were Prdm4 (PR domain 4) > Ube4a (Ubiquitin-Conjugating Enzyme 4a) > Enox2 (Ecto-NOX Disulfide-Thiol Exchanger 2) > Ube2d2 (Ubiquitin-conjugating enzyme E2D 2) > Actb (Actin beta). We therefore recommend using at least four of the aforementioned HKG for normalisation of endometrial tissues taken from patients with RM and RIF.

  13. Live Imaging of Type I Collagen Assembly Dynamics in Osteoblasts Stably Expressing GFP and mCherry-Tagged Collagen Constructs.

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    Lu, Yongbo; Kamel-El Sayed, Suzan A; Wang, Kun; Tiede-Lewis, LeAnn M; Grillo, Michael A; Veno, Patricia A; Dusevich, Vladimir; Phillips, Charlotte L; Bonewald, Lynda F; Dallas, Sarah L

    2018-02-20

    Type I collagen is the most abundant extracellular matrix protein in bone and other connective tissues and plays key roles in normal and pathological bone formation as well as in connective tissue disorders and fibrosis. Although much is known about the collagen biosynthetic pathway and its regulatory steps, the mechanisms by which it is assembled extracellularly are less clear. We have generated GFPtpz and mCherry-tagged collagen fusion constructs for live imaging of type I collagen assembly by replacing the α2(I)-procollagen N-terminal propeptide with GFPtpz or mCherry. These novel imaging probes were stably transfected into MLO-A5 osteoblast-like cells and fibronectin-null mouse embryonic fibroblasts (FN-null-MEFs) and used for imaging type I collagen assembly dynamics and its dependence on fibronectin. Both fusion proteins co-precipitated with α1(I)-collagen and remained intracellular without ascorbate but were assembled into α1(I) collagen-containing extracellular fibrils in the presence of ascorbate. Immunogold-EM confirmed their ultrastuctural localization in banded collagen fibrils. Live cell imaging in stably transfected MLO-A5 cells revealed the highly dynamic nature of collagen assembly and showed that during assembly the fibril networks are continually stretched and contracted due to the underlying cell motion. We also observed that cell-generated forces can physically reshape the collagen fibrils. Using co-cultures of mCherry- and GFPtpz-collagen expressing cells, we show that multiple cells contribute collagen to form collagen fiber bundles. Immuno-EM further showed that individual collagen fibrils can receive contributions of collagen from more than one cell. Live cell imaging in FN-null-MEFs expressing GFPtpz-collagen showed that collagen assembly was both dependent upon and dynamically integrated with fibronectin assembly. These GFP-collagen fusion constructs provide a powerful tool for imaging collagen in living cells and have revealed novel

  14. Characteristics of stably expressed human dopamine D1a and D1b receptors: atypical behavior of the dopamine D1b receptor

    DEFF Research Database (Denmark)

    Pedersen, U B; Norby, B; Jensen, Anders A.

    1994-01-01

    Human dopamine D1a and D1b receptors were stably expressed in Baby Hamster Kidney (BHK) or Chinese Hamster Ovary (CHO) cells. [3H]SCH23390 saturation experiments indicated the presence of only a single binding site in the D1a expressing cell line with a Kd of 0.5 nM. In D1b expressing cell lines...

  15. Harnessing gene expression networks to prioritize candidate epileptic encephalopathy genes.

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    Oliver, Karen L; Lukic, Vesna; Thorne, Natalie P; Berkovic, Samuel F; Scheffer, Ingrid E; Bahlo, Melanie

    2014-01-01

    We apply a novel gene expression network analysis to a cohort of 182 recently reported candidate Epileptic Encephalopathy genes to identify those most likely to be true Epileptic Encephalopathy genes. These candidate genes were identified as having single variants of likely pathogenic significance discovered in a large-scale massively parallel sequencing study. Candidate Epileptic Encephalopathy genes were prioritized according to their co-expression with 29 known Epileptic Encephalopathy genes. We utilized developing brain and adult brain gene expression data from the Allen Human Brain Atlas (AHBA) and compared this to data from Celsius: a large, heterogeneous gene expression data warehouse. We show replicable prioritization results using these three independent gene expression resources, two of which are brain-specific, with small sample size, and the third derived from a heterogeneous collection of tissues with large sample size. Of the nineteen genes that we predicted with the highest likelihood to be true Epileptic Encephalopathy genes, two (GNAO1 and GRIN2B) have recently been independently reported and confirmed. We compare our results to those produced by an established in silico prioritization approach called Endeavour, and finally present gene expression networks for the known and candidate Epileptic Encephalopathy genes. This highlights sub-networks of gene expression, particularly in the network derived from the adult AHBA gene expression dataset. These networks give clues to the likely biological interactions between Epileptic Encephalopathy genes, potentially highlighting underlying mechanisms and avenues for therapeutic targets.

  16. NF-kappa B activity in T cells stably expressing the Tax protein of human T cell lymphotropic virus type I

    International Nuclear Information System (INIS)

    Lacoste, J.; Cohen, L.; Hiscott, J.

    1991-01-01

    The effect of constitutive Tax expression on the interaction of NF-κ B with its recognition sequence and on NF-κ B-dependent gene expression was examined in T lymphoid Jurkat cell lines (19D and 9J) stably transformed with a Tax expression vector. Tax expressing T cell lines contained a constitutive level of NF-κ B binding activity, detectable by mobility shift assay and uv cross-linking using a palindromic NF-κ B probe homologous to the interferon beta PRDII site. In Jurkat and NC2.10 induction with phorbol esters resulted in the appearance of new DNA binding proteins of 85, 75, and 54 kDa, whereas in Tax expressing cells the 85-kDa protein and a 92-kDa DNA binding protein were constitutively induced. Expression of Tax protein in 19D and 9J resulted in transcription of the endogenous NF-kappa B-dependent granulocyte-macrophage colony stimulating factor gene and increased basal level expression of transfected NF-kappa B-regulated promoters. Nonetheless transcription of both the endogenous and the transfected gene was inducible by PMA treatment. Tax expression in Jurkat T cells may alter the stoichiometry of NF-kappa B DNA binding proteins and thus change the expression of NF-kappa B-regulated promoters

  17. SPARC Expression Is Selectively Suppressed in Tumor Initiating Urospheres Isolated from As+3- and Cd+2-Transformed Human Urothelial Cells (UROtsa Stably Transfected with SPARC.

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    Andrea Slusser-Nore

    Full Text Available This laboratory previously analyzed the expression of SPARC in the parental UROtsa cells, their arsenite (As(+3 and cadmium (Cd(+2-transformed cell lines, and tumor transplants generated from the transformed cells. It was demonstrated that SPARC expression was down-regulated to background levels in Cd(+2-and As(+3-transformed UROtsa cells and tumor transplants compared to parental cells. In the present study, the transformed cell lines were stably transfected with a SPARC expression vector to determine the effect of SPARC expression on the ability of the cells to form tumors in immune-compromised mice.Real time PCR, western blotting, immunohistochemistry, and immunofluorescence were used to define the expression of SPARC in the As(+3-and Cd(+2-transformed cell lines, and urospheres isolated from these cell lines, following their stable transfection with an expression vector containing the SPARC open reading frame (ORF. Transplantation of the cultured cells into immune-compromised mice by subcutaneous injection was used to assess the effect of SPARC expression on tumors generated from the above cell lines and urospheres.It was shown that the As(+3-and Cd(+2-transformed UROtsa cells could undergo stable transfection with a SPARC expression vector and that the transfected cells expressed both SPARC mRNA and secreted protein. Tumors formed from these SPARC-transfected cells were shown to have no expression of SPARC. Urospheres isolated from cultures of the SPARC-transfected As(+3-and Cd(+2-transformed cell lines were shown to have only background expression of SPARC. Urospheres from both the non-transfected and SPARC-transfected cell lines were tumorigenic and thus fit the definition for a population of tumor initiating cells.Tumor initiating cells isolated from SPARC-transfected As(+3-and Cd(+2-transformed cell lines have an inherent mechanism to suppress the expression of SPARC mRNA.

  18. Generation of induced pluripotent stem cells (iPSCs) stably expressing CRISPR-based synergistic activation mediator (SAM)

    DEFF Research Database (Denmark)

    Xiong, Kai; Zhou, Yan; Hyttel, Poul

    2016-01-01

    Human fibroblasts were engineered to express the CRISPR-based synergistic activation mediator (SAM) complex: dCas9-VP64 and MS2-P65-HSF1. Two induced pluripotent stem cells (iPSCs) clones expressing SAM were established by transducing these fibroblasts with lentivirus expressing OCT4, SOX2, KLF4...... a novel, useful tool to investigate genetic regulation of stem cell proliferation and differentiation through CRISPR-mediated activation of endogenous genes....

  19. Characterization of Caco-2 cells stably expressing the protein-based zinc probe eCalwy-5 as a model system for investigating intestinal zinc transport.

    Science.gov (United States)

    Maares, Maria; Keil, Claudia; Thomsen, Susanne; Günzel, Dorothee; Wiesner, Burkhard; Haase, Hajo

    2018-01-29

    Intestinal zinc resorption, in particular its regulation and mechanisms, are not yet fully understood. Suitable intestinal cell models are needed to investigate zinc uptake kinetics and the role of labile zinc in enterocytes in vitro. Therefore, a Caco-2 cell clone was produced, stably expressing the genetically encoded zinc biosensor eCalwy-5. The aim of the present study was to reassure the presence of characteristic enterocyte-specific properties in the Caco-2-eCalwy clone. Comparison of Caco-2-WT and Caco-2-eCalwy cells revealed only slight differences regarding subcellular localization of the tight junction protein occludin and alkaline phosphatase activity, which did not affect basic integrity of the intestinal barrier or the characteristic brush border membrane morphology. Furthermore, introduction of the additional zinc-binding protein in Caco-2 cells did not alter mRNA expression of the major intestinal zinc transporters (zip4, zip5, znt-1 and znt-5), but increased metallothionein 1a-expression and cellular resistance to higher zinc concentrations. Moreover, this study examines the effect of sensor expression level on its saturation with zinc. Fluorescence cell imaging indicated considerable intercellular heterogeneity in biosensor-expression. However, FRET-measurements confirmed that these differences in expression levels have no effect on fractional zinc-saturation of the probe. Copyright © 2018 Elsevier GmbH. All rights reserved.

  20. Efficient propagation of progressive multifocal leukoencephalopathy-type JC virus in COS-7-derived cell lines stably expressing Tat protein of human immunodeficiency virus type 1.

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    Nukuzuma, Souichi; Nakamichi, Kazuo; Kameoka, Masanori; Sugiura, Shigeki; Nukuzuma, Chiyoko; Miyoshi, Isao; Takegami, Tsutomu

    2010-12-01

    The high incidence of progressive multifocal leukoencephalopathy (PML) in AIDS patients compared with many other immunosuppressive diseases suggests that HIV-1 infection is strictly related to the activation of JC virus (JCV) propagation. In this report, propagation of PML-type JCV in COS-7-derived cell lines stably expressing HIV-1 Tat (COS-tat cells) has been examined. In COS-tat cells, production of viral particles and replication of genomic DNA were markedly increased compared to COS-7 cells, as judged by HA and real-time PCR analyses. These results demonstrate that COS-tat cells provide a useful model system for studying HIV-1 Tat-mediated propagation of PML-type JCV. © 2010 The Societies and Blackwell Publishing Asia Pty Ltd.

  1. Characterization of calcium signals in human induced pluripotent stem cell-derived dentate gyrus neuronal progenitors and mature neurons, stably expressing an advanced calcium indicator protein.

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    Vőfély, Gergő; Berecz, Tünde; Szabó, Eszter; Szebényi, Kornélia; Hathy, Edit; Orbán, Tamás I; Sarkadi, Balázs; Homolya, László; Marchetto, Maria C; Réthelyi, János M; Apáti, Ágota

    2018-04-01

    Pluripotent stem cell derived human neuronal progenitor cells (hPSC-NPCs) and their mature neuronal cell culture derivatives may efficiently be used for central nervous system (CNS) drug screening, including the investigation of ligand-induced calcium signalization. We have established hippocampal NPC cultures derived from human induced PSCs, which were previously generated by non-integrating Sendai virus reprogramming. Using established protocols these NPCs were differentiated into hippocampal dentate gyrus neurons. In order to study calcium signaling without the need of dye loading, we have stably expressed an advanced calcium indicator protein (GCaMP6fast) in the NPCs using the Sleeping Beauty transposon system. We observed no significant effects of the long-term GCaMP6 expression on NPC morphology, gene expression pattern or neural differentiation capacity. In order to compare the functional properties of GCaMP6-expressing neural cells and the corresponding parental cells loaded with calcium indicator dye Fluo-4, a detailed characterization of calcium signals was performed. We found that the calcium signals induced by ATP, glutamate, LPA, or proteases - were similar in these two systems. Moreover, the presence of the calcium indicator protein allowed for a sensitive, repeatable detection of changes in calcium signaling during the process of neurogenesis and neuronal maturation. Copyright © 2018 Elsevier Inc. All rights reserved.

  2. Expression of calmodulin and calmodulin binding proteins in rat fibroblasts stably transfected with protein kinase C and oncogenes

    DEFF Research Database (Denmark)

    Ye, Q; Wei, Y; Fischer, R

    1997-01-01

    Molecular mechanisms leading to elevated calmodulin (CaM) expression in cancer have not yet been discovered. We have quantitated the levels of transcripts derived from all three CaM genes in a variety of the same origin rat fibroblasts transformed with oncogenes in combination with gene for prote...

  3. Expression of calmodulin and calmodulin binding proteins in rat fibroblasts stably transfected with protein kinase C and oncogenes

    DEFF Research Database (Denmark)

    Ye, Q; Wei, Y; Fischer, R

    1997-01-01

    Molecular mechanisms leading to elevated calmodulin (CaM) expression in cancer have not yet been discovered. We have quantitated the levels of transcripts derived from all three CaM genes in a variety of the same origin rat fibroblasts transformed with oncogenes in combination with gene for protein...

  4. Characterization of a human cell line stably over-expressing the candidate oncogene, dual specificity phosphatase 12.

    Directory of Open Access Journals (Sweden)

    Erica L Cain

    2011-04-01

    Full Text Available Analysis of chromosomal rearrangements within primary tumors has been influential in the identification of novel oncogenes. Identification of the "driver" gene(s within cancer-derived amplicons is, however, hampered by the fact that most amplicons contain many gene products. Amplification of 1q21-1q23 is strongly associated with liposarcomas and microarray-based comparative genomic hybridization narrowed down the likely candidate oncogenes to two: the activating transcription factor 6 (atf6 and the dual specificity phosphatase 12 (dusp12. While atf6 is an established transcriptional regulator of the unfolded protein response, the potential role of dusp12 in cancer remains uncharacterized.To evaluate the oncogenic potential of dusp12, we established stable cell lines that ectopically over-express dusp12 in isolation and determined whether this cell line acquired properties frequently associated with transformed cells. Here, we demonstrate that cells over-expressing dusp12 display increased cell motility and resistance to apoptosis. Additionally, over-expression of dusp12 promoted increased expression of the c-met proto-oncogene and the collagen and laminin receptor intergrin alpha 1 (itga1 which is implicated in metastasis.Collectively, these results suggest that dusp12 is oncologically relevant and exposes a potential association between dusp12 and established oncogenes that could be therapeutically targeted.

  5. Characteristics of stably expressed human dopamine D1a and D1b receptors: atypical behavior of the dopamine D1b receptor

    DEFF Research Database (Denmark)

    Pedersen, U B; Norby, B; Jensen, Anders A.

    1994-01-01

    Human dopamine D1a and D1b receptors were stably expressed in Baby Hamster Kidney (BHK) or Chinese Hamster Ovary (CHO) cells. [3H]SCH23390 saturation experiments indicated the presence of only a single binding site in the D1a expressing cell line with a Kd of 0.5 nM. In D1b expressing cell lines......, two binding sites were observed with Kd values of 0.5 and 5 nM in CHO cells and 0.05 and 1.6 nM in BHK cells, respectively. Neither of the receptors affected Ca2+ metabolism whereas they both were coupled in a stimulatory fashion to adenylyl cyclase. The pharmacological profile of both the D1a and D1b...... for these receptors. Besides SCH 23390, only NNC 112, fluphenazine and bulbocapnine were able to discriminate between the two states of the D1b receptor. In case of the D1a receptor, the Ki values obtained in binding experiments were very similar to Ki values obtained from inhibition of dopamine stimulated adenylyl...

  6. Persistent human cardiac Na+ currents in stably transfected mammalian cells: Robust expression and distinct open-channel selectivity among Class 1 antiarrhythmics.

    Science.gov (United States)

    Wang, Ging Kuo; Russell, Gabriella; Wang, Sho-Ya

    2013-01-01

    Miniature persistent late Na(+) currents in cardiomyocytes have been linked to arrhythmias and sudden death. The goals of this study are to establish a stable cell line expressing robust persistent cardiac Na(+) currents and to test Class 1 antiarrhythmic drugs for selective action against resting and open states. After transient transfection of an inactivation-deficient human cardiac Na(+) channel clone (hNav1.5-CW with L409C/A410W double mutations), transfected mammalian HEK293 cells were treated with 1 mg/ml G-418. Individual G-418-resistant colonies were isolated using glass cylinders. One colony with high expression of persistent Na(+) currents was subjected to a second colony selection. Cells from this colony remained stable in expressing robust peak Na(+) currents of 996 ± 173 pA/pF at +50 mV (n = 20). Persistent late Na(+) currents in these cells were clearly visible during a 4-second depolarizing pulse albeit decayed slowly. This slow decay is likely due to slow inactivation of Na(+) channels and could be largely eliminated by 5 μM batrachotoxin. Peak cardiac hNav1.5-CW Na(+) currents were blocked by tetrodotoxin with an IC(50) value of 2.27 ± 0.08 μM (n = 6). At clinic relevant concentrations, Class 1 antiarrhythmics are much more selective in blocking persistent late Na(+) currents than their peak counterparts, with a selectivity ratio ranging from 80.6 (flecainide) to 3 (disopyramide). We conclude that (1) Class 1 antiarrhythmics differ widely in their resting- vs. open-channel selectivity, and (2) stably transfected HEK293 cells expressing large persistent hNav1.5-CW Na(+) currents are suitable for studying as well as screening potent open-channel blockers.

  7. Pharmacological and functional characterisation of the wild-type and site-directed mutants of the human H1 histamine receptor stably expressed in CHO cells.

    Science.gov (United States)

    Moguilevsky, N; Varsalona, F; Guillaume, J P; Noyer, M; Gillard, M; Daliers, J; Henichart, J P; Bollen, A

    1995-01-01

    A cDNA clone for the human histamine H1 receptor was isolated from a lung cDNA library and stably expressed in CHO cells. The recombinant receptor protein present in the cell membranes, displayed the functional and binding characteristics of histamine H1 receptors. Mutation of Ser155 to Ala in the fourth transmembrane domain did not significantly change the affinity of the receptor for histamine and H1 antagonists. However, mutation of the fifth transmembrane Asn198 to Ala resulted in a dramatic decrease of the affinity for histamine binding, and for the histamine-induced polyphosphoinositides breakdown, whereas the affinity towards antagonists was not significantly modified. In addition, mutation of another fifth transmembrane amino acid, Thr194 to Ala also diminished, but to a lesser extent, the affinity for histamine. These data led us to propose a molecular model for histamine interaction with the human H1 receptor. In this model, the amide moiety of Asn198 and the hydroxyl group of Thr194 are involved in hydrogen bonding with the nitrogen atoms of the imidazole ring of histamine. Moreover, mutation of Thr194 to Ala demonstrated that this residue is responsible for the discrimination between enantiomers of cetirizine.

  8. Phenylpropanoids from cinnamon bark reduced β-amyloid production by the inhibition of β-secretase in Chinese hamster ovarian cells stably expressing amyloid precursor protein.

    Science.gov (United States)

    Kang, Yu Jeong; Seo, Dae-Gun; Park, So-Young

    2016-11-01

    β-Amyloid (Aβ) is a substance of Alzheimer disease (AD), which is generated via the amyloidogenic pathway from amyloid precursor protein (APP) by β-secretase and γ-secretase. Inhibition of Aβ production is a potential therapeutic approach to AD. Thus, we tested the hypothesis that cinnamon bark (Cinnamomi Cortex Spissus), the dried bark of Cinnamomum cassia Blume (Lauraceae), and its constituents are beneficial to AD. The methanol extract of cinnamon bark efficiently reduced Aβ40 production in Chinese hamster ovarian (CHO) cells stably expressing APP as determined by enzyme-linked immunosorbent assay. Bioassay-guided isolation of cinnamon bark extract was carried out using open column chromatography and high-performance liquid chromatography, and the following 6 phenylpropanoids were isolated: syringaresinol (1); medioresinol (2); coumarin (3); 2-hydroxycinnamaldehyde (4); cryptamygin A (5); and 3',5,7-trimethoxy epicatechin (6). Among these, 4 μg/mL medioresinol and cryptamygin A reduced Aβ40 production by 50% and 60%, respectively, compared with dimethyl sulfoxide-treated control cells. The IC 50 values of medioresinol and cryptamygin A for the inhibition of Aβ40 production were 10.8 and 8.2 μg/mL, respectively. Furthermore, treatment of APP-CHO cells with either compound decreased the amount of β-secretase and sAPPβ (the proteolytic fragment of APP catalyzed by β-secretase). These results suggest that the antiamyloidogenic activity of cinnamon bark extract was exerted by medioresinol and cryptamygin A via a reduction in the amount of β-secretase. The extract of cinnamon bark contains potentially valuable antiamyloidogenic agents for the prevention and treatment of AD. Copyright © 2016 Elsevier Inc. All rights reserved.

  9. Nicotine Component of Cigarette Smoke Extract (CSE) Decreases the Cytotoxicity of CSE in BEAS-2B Cells Stably Expressing Human Cytochrome P450 2A13.

    Science.gov (United States)

    Ji, Minghui; Zhang, Yudong; Li, Na; Wang, Chao; Xia, Rong; Zhang, Zhan; Wang, Shou-Lin

    2017-10-13

    Cytochrome P450 2A13 (CYP2A13), an extrahepatic enzyme mainly expressed in the human respiratory system, has been reported to mediate the metabolism and toxicity of cigarette smoke. We previously found that nicotine inhibited 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanone (NNK) metabolism by CYP2A13, but its influence on other components of cigarette smoke remains unclear. The nicotine component of cigarette smoke extract (CSE) was separated, purified, and identified using high-performance liquid chromatography (HPLC) and ultra-performance liquid chromatography tandem mass spectrometry (UPLC-MS/MS), splitting CSE into a nicotine section (CSE-N) and nicotine-free section (CSE-O). Cell viability and apoptosis by Cell Counting Kit-8 (CCK-8) and flow cytometry assays were conducted on immortalized human bronchial epithelial (BEAS-2B) cells stably expressing CYP2A13 (B-2A13) or vector (B-V), respectively. Interestingly, CSE and CSE-O were toxic to BEAS-2B cells whereas CSE-N showed less cytotoxicity. CSE-O was more toxic to B-2A13 cells than to B-V cells (IC 50 of 2.49% vs. 7.06%), which was flatted by 8-methoxypsoralen (8-MOP), a CYP inhibitor. CSE-O rather than CSE or CSE-N increased apoptosis of B-2A13 cells rather than B-V cells. Accordingly, compared to CSE-N and CSE, CSE-O significantly changed the expression of three pairs of pro- and anti-apoptotic proteins, Bcl-2 Associated X Protein/B cell lymphoma-2 (Bax/Bcl-2), Cleaved Poly (Adenosine Diphosphate-Ribose) Polymerase/Poly (Adenosine Diphosphate-Ribose) Polymerase (C-PARP/PARP), and C-caspase-3/caspase-3, in B-2A13 cells. In addition, recombination of CSE-N and CSE-O (CSE-O/N) showed similar cytotoxicity and apoptosis to the original CSE. These results demonstrate that the nicotine component decreases the metabolic activation of CYP2A13 to CSE and aids in understanding the critical role of CYP2A13 in human respiratory diseases caused by cigarette smoking.

  10. [Selection and construction of cell line stably expressing survivin gene in lower level through eukaryotic plasmid vector of shRNA].

    Science.gov (United States)

    Wang, Wen-Xia; Sun, Shan-Zhen; Song, Ying

    2008-06-01

    To construct a short hairpin RNA(shRNA) interference expression plasmid vector of survivin gene, transfect tongue squamous cell carcinoma line Tca8113 which expressed survivin gene in a high level, and choose the cells whose survivin gene were suppressed significantly. Two pairs of oligonucleotide sequences specific for survivin gene were designed and synthesized, and cloned into pSilencer-2.1U6-neo plasmid. The recombinant plasmids (named PS1 and PS2) were amplified in Ecoli. DH5alpha was identified by restriction digestion, PCR and sequencing. The vectors were transfected into Tca8113 cells with lipofectamine 2000. After selection with G418, the stable cell clones were attained. Survivn expression was assayed with real-time quantitative PCR and Western blotting. SAS8.0 software package was used for Student t test. Two vectors were constructed successfully and stable cell clones with PS1 or PS2 plasmid were obtained. As compared with those of control, survivin expression of transfected cell with PS1 or PS2 in mRNA level was significantly suppressed (P<0.05). In protein level, only those of transfected cell with PS2 was significantly suppressed (P<0.01). The shRNA interference expression plasmid vectors of survivin gene are successfully constructed, and Tca8113 cells which express survivin gene in a stable lower level are attained, which enable us to carry out further research on gene therapy of oral squamous cell carcinoma. Supported by National Natural Science Foundation of China (Grant No.30572056).

  11. GH induced lipolysis stimulation in 3T3-L1 adipocytes stably expressing hGHR: analysis on signaling pathway and activity of 20K hGH.

    Science.gov (United States)

    Asada, N; Takahashi, Y; Wada, M; Naito, N; Uchida, H; Ikeda, M; Honjo, M

    2000-04-25

    We have constructed a cell line of 3T3-L1 which can efficiently express human GHR (3T3-L1-hGHR) after differentiation to adipocytes. The expressed hGHR was detected as two bands with approximate molecular sizes of 120K by Western analysis using hGHR specific monoclonal antibody. Maximum lipolytic activity induced by hGH in the 3T3-L1-hGHR was enhanced 10-fold as compared to that in 3T3-L1, suggesting that expressed hGHR is functionally active. Comparative analysis using bGH and hGH revealed that 70% of lipolysis stimulation by 1-10 ng/ml hGH could be attributed to hGHR-mediated response. Analyses on inhibition and phosphorylation of signaling molecules suggested that GH-induced lipolysis stimulation is dependent on gene expression and not mediated through PKA-, PKC-, PLA-, PLC-, nor MAPK-pathway but possibly through JAK-STATs pathway. Duration of STAT5 activation by hGH continued up to 48 h. We also revealed that 22 K hGH isoform, 20K hGH which has been reported as a weaker agonist for GH-induced lipolysis stimulation, possesses equipotent activity and shows stronger action in the presence of hGHBP as compared to 22 K hGH. Taken together we conclude that the hGH-induced lipolysis was not mediated through MAP-, PKA-, PKC-, nor PLA-pathway but might be mediated through STAT pathway and that 20K hGH might show higher lipolytic activity than 22 K hGH in adipose tissue that produces a large amount of GHBP.

  12. Biologically active, magnICON®-expressed EPO-Fc from stably transformed Nicotiana benthamiana plants presenting tetra-antennary N-glycan structures.

    Science.gov (United States)

    Nagels, Bieke; Van Damme, Els J M; Callewaert, Nico; Zabeau, Lennart; Tavernier, Jan; Delanghe, Joris R; Boets, Annemie; Castilho, Alexandra; Weterings, Koen

    2012-08-31

    In the past two decades plants have emerged as a valuable alternative for the production of pharmaceutical proteins. Since N-glycosylation influences functionality and stability of therapeutic proteins, the plant N-glycosylation pathway should be humanized. Here, we report the transient magnICON(®) expression of the erythropoietin fusion protein (EPO-Fc) in Nicotiana benthamiana plants that produce multi-antennary N-glycans without the plant-specific β1,2-xylose and α1,3-fucose residues in a stable manner (Nagels et al., 2011). The EPO-Fc fusion protein consists of EPO with a C-terminal-linked IgG-Fc domain and is used for pulmonary delivery of recombinant EPO to patients (Bitonti et al., 2004). Plant expressed EPO-Fc was quantified using a paramagnetic-particle chemiluminescent immunoassay and shown to be active in vitro via receptor binding experiments in HEK293T cells. Mass spectrometry-based N-glycan analysis confirmed the presence of multi-antennary N-glycans on plant-expressed EPO-Fc. The described research is the next step towards the development of a production platform for pharmaceutical proteins in plants. Copyright © 2012 Elsevier B.V. All rights reserved.

  13. Harnessing inequality

    Science.gov (United States)

    Eliazar, Iddo

    2016-09-01

    Living in the era of "big-data" information, we are ubiquitously inundated by overabundances of sizes-non-negative numerical values representing count, score, length, area, volume, duration, mass, energy, etc. Datasets of sizes display numerous types of statistical variability that are commonly quantified either by the standard deviation, or by the Boltzmann-Gibbs-Shannon entropy. The standard deviation measures the sizes' Euclidean divergence from their mean, the Boltzmann-Gibbs-Shannon entropy measures the sizes' informational divergence from the benchmark of pure determinism, and both these gauges are one-dimensional. In this paper we overview a methodology that harnesses inequality in order to quantify statistical variability. The methodology follows a socioeconomic approach of measuring the sizes' inequality-their divergence from the benchmark of pure egalitarianism-and yields frameworks that gauge statistical variability in a multi-dimensional fashion. The aim of this overview is to serve both researchers and practitioners as a crash-introduction to the "harnessing inequality" methodology, and as a crash-manual to the implementation of this methodology.

  14. Molecular Imaging of Human Embryonic Stem Cells Stably Expressing Human PET Reporter Genes After Zinc Finger Nuclease-Mediated Genome Editing.

    Science.gov (United States)

    Wolfs, Esther; Holvoet, Bryan; Ordovas, Laura; Breuls, Natacha; Helsen, Nicky; Schönberger, Matthias; Raitano, Susanna; Struys, Tom; Vanbilloen, Bert; Casteels, Cindy; Sampaolesi, Maurilio; Van Laere, Koen; Lambrichts, Ivo; Verfaillie, Catherine M; Deroose, Christophe M

    2017-10-01

    Molecular imaging is indispensable for determining the fate and persistence of engrafted stem cells. Standard strategies for transgene induction involve the use of viral vectors prone to silencing and insertional mutagenesis or the use of nonhuman genes. Methods: We used zinc finger nucleases to induce stable expression of human imaging reporter genes into the safe-harbor locus adeno-associated virus integration site 1 in human embryonic stem cells. Plasmids were generated carrying reporter genes for fluorescence, bioluminescence imaging, and human PET reporter genes. Results: In vitro assays confirmed their functionality, and embryonic stem cells retained differentiation capacity. Teratoma formation assays were performed, and tumors were imaged over time with PET and bioluminescence imaging. Conclusion: This study demonstrates the application of genome editing for targeted integration of human imaging reporter genes in human embryonic stem cells for long-term molecular imaging. © 2017 by the Society of Nuclear Medicine and Molecular Imaging.

  15. Harnessing gas

    International Nuclear Information System (INIS)

    1998-01-01

    Nigeria produces almost two million barrels of oil a day from its oil fields in the Niger Delta area. Most of the oil comes from reservoirs containing gas, which is produced with the oil. This associated gas is separated from the oil at flow stations and more than 95 per cent of it is flared-currently a total of some two billion standard cubic feet per day (scf/d), which is estimated to be about a quarter of the gas the world flares and vents. The energy available from Nigeria's flared gas is prodigious, equivalent to one quarter of France's gas requirements. The company recognises that flaring wastes a valuable resource and is environmentally damaging. It aims to stop necessary flaring as soon as possible through a series of projects to harness or conserve this gas. Several gas gathering and conservation projects are already underway in response to emerging markets while other plans await new markets. The company is committed to reduce gas flaring as soon as is feasible to the minimum needed to maintain safe operations. But why are solutions being found only now? why has Nigeria been flaring gas for so long? These question lie at the crux of the debate about Nigeria and gas flaring and the answers, which continue to have a major impact on associated gas development today, are rooted in history, economics and geography

  16. Development of an Immunoperoxidase Monolayer Assay for the Detection of Antibodies against Peste des Petits Ruminants Virus Based on BHK-21 Cell Line Stably Expressing the Goat Signaling Lymphocyte Activation Molecule.

    Directory of Open Access Journals (Sweden)

    Jialin Zhang

    Full Text Available From 2013 to 2015, peste des petits ruminants (PPR broke out in more than half of the provinces of China; thus, the application and development of diagnostic methods are very important for the control of PPR. Here, an immunoperoxidase monolayer assay (IPMA was developed to detect antibodies against PPR. However, during IPMA development, we found that Vero cells were not the appropriate choice because staining results were not easily observed. Therefore, we first established a baby hamster kidney-goat signaling lymphocyte activation molecule (BHK-SLAM cell line that could stably express goat SLAM for at least 20 generations. Compared with Vero cells, the PPR-mediated cytopathic effect occurred earlier in BHK-SLAM cells, and large syncytia appeared after virus infection. Based on this cell line and recombinant PPR virus expressing the green fluorescent protein (GFP (rPPRV-GFP, an IPMA for PPR diagnosis was developed. One hundred and ninety-eight PPR serum samples from goats or sheep were tested by the IPMA and virus neutralization test (VNT. Compared with the VNT, the sensitivity and specificity of the IPMA were 91% and 100%, respectively, and the coincidence rate of the two methods was 95.5%. The IPMA assay could be completed in 4 h, compared with more than 6 d for the VNT using rPPRV-GFP, and it is easily performed, as the staining results can be observed under a microscope. Additionally, unlike the VNT, the IPMA does not require antigen purification, which will reduce its cost. In conclusion, the established IPMA will be an alternative method that replaces the VNT for detecting antibodies against PPRV in the field.

  17. Har erhvervsuddannelserne en fremtid?

    DEFF Research Database (Denmark)

    Jørgensen, Christian Helms

    2012-01-01

    Har erhvervsuddannelserne en fremtid? Eller mere præcist: Har de en fremtid med den grundform som nu har fungeret i over 50 år? Det vil sige som et separat system baseret på vekseluddannelser, hvor hovedparten af uddannelserne foregår i praktik, og som i vid udstrækning styres af arbejdsmarkedets...

  18. Stably-stratified wall-bounded turbulence

    Science.gov (United States)

    Hadi Sichani, Pejman; Zonta, Francesco; Obabko, Aleksandr; Soldati, Alfredo

    2017-11-01

    Stably-stratified (bottom-up cooling) turbulent flows are encountered in a number of industrial applications, environmental processes and geophysical flows. Turbulent entrainment and mixing across density interfaces in terrestrial water bodies (oceans, lakes and rivers) and in industrial heat transfer equipments are just some important examples of stably-stratified flows. In this work we use Direct Numerical Simulation to investigate the fundamental physics of stably-stratified channel turbulence under Boussinesq and Non-Oberbeck-Boussinesq (NOB) conditions. Compared to the neutrally-buoyant case, in the stably-stratified case active turbulence survives only in the near-wall region and coexists with internal gravity waves (IGW) moving in the core region of the channel. This induces a general suppression of turbulence levels, momentum and buoyancy fluxes. Our results show also that NOB effects may be important when the flow is subject to large temperature gradients. The most striking feature observed in case of NOB conditions is the generation of a strong flow asymmetry with possible local flow laminarization in the near wall region.

  19. Large eddy simulation of stably stratified turbulence

    International Nuclear Information System (INIS)

    Shen Zhi; Zhang Zhaoshun; Cui Guixiang; Xu Chunxiao

    2011-01-01

    Stably stratified turbulence is a common phenomenon in atmosphere and ocean. In this paper the large eddy simulation is utilized for investigating homogeneous stably stratified turbulence numerically at Reynolds number Re = uL/v = 10 2 ∼10 3 and Froude number Fr = u/NL = 10 −2 ∼10 0 in which u is root mean square of velocity fluctuations, L is integral scale and N is Brunt-Vaïsälä frequency. Three sets of computation cases are designed with different initial conditions, namely isotropic turbulence, Taylor Green vortex and internal waves, to investigate the statistical properties from different origins. The computed horizontal and vertical energy spectra are consistent with observation in atmosphere and ocean when the composite parameter ReFr 2 is greater than O(1). It has also been found in this paper that the stratification turbulence can be developed under different initial velocity conditions and the internal wave energy is dominated in the developed stably stratified turbulence.

  20. Helle har din ryg

    DEFF Research Database (Denmark)

    Kock, Christian Erik J

    2015-01-01

    Thornings 1. maj-tale er ikke noget mesterværk af en levende, inciterende, inviterende tekst. Her er stadig for mange slatne sætninger og metaforer, der har overskredet sidste muggenhedsdato. Men talen her har bestemt sine takter – af noget, som er alt for sjældent i dansk politisk diskurs: gnist...

  1. Harnessing vision for computation.

    Science.gov (United States)

    Changizi, Mark

    2008-01-01

    Might it be possible to harness the visual system to carry out artificial computations, somewhat akin to how DNA has been harnessed to carry out computation? I provide the beginnings of a research programme attempting to do this. In particular, new techniques are described for building 'visual circuits' (or 'visual software') using wire, NOT, OR, and AND gates in a visual 6modality such that our visual system acts as 'visual hardware' computing the circuit, and generating a resultant perception which is the output.

  2. Har vi en virkelighedssans?

    DEFF Research Database (Denmark)

    Trettvik, Johan

    2013-01-01

    Vores teoretiske forhold til virkeligheden er meget omdiskuteret, og særlig spørgsmålet om, hvorvidt vores viden afspejler noget virkeligt, er centralt i diskussionen af begrebet om virkelighed. I psykologien har det sat sine spor ved enten en reduktionistisk holdning til sit genstandsfelt eller ...

  3. Har Gobind Khorana

    Indian Academy of Sciences (India)

    Home; Journals; Resonance – Journal of Science Education; Volume 17; Issue 12. Har Gobind Khorana - Early Years in Science and Transition from Chemistry to Chemical Biology. Uttam L Rajbhandary. General Article Volume 17 Issue 12 December 2012 pp 1125-1135 ...

  4. Kumar, Prof. Har Darshan

    Indian Academy of Sciences (India)

    Fellow Profile. Elected: 1975 Section: Plant Sciences. Kumar, Prof. Har Darshan Ph.D. (London), FNA, FNASc. Date of birth: 25 February 1934. Specialization: Algology, Microbial Biotechnology, Genetics, Ecology and Environment Address: Mrigtrishna, B.32/32, H.K. 214, Saketnagar Colony, Naria, Varanasi 221 005, U.P.

  5. Har Gobind Khorana

    Indian Academy of Sciences (India)

    IAS Admin

    This article provides a brief summary of Har Gobind. Khorana's early years in science including his years as an independent investigator at the British Columbia Research. Council in Vancouver, Canada. It also includes interesting anecdotes about his career and about him as a scientist based on his own writings, including ...

  6. Khorana, Prof. Har Gobind

    Indian Academy of Sciences (India)

    Elected: 1976 Honorary. Khorana, Prof. Har Gobind Nobel Laureate (Medicine) - 1968. Date of birth: 9 January 1922. Date of death: 9 November 2011. Last known address: Alfred P. Sloan Professor of, Biology and Chemistry, Massachusetts Institute of, Technology, 77 Massachusetts Ave., Cambridge, MA 02139, U.S.A..

  7. Generation of H9 T-cells stably expressing a membrane-bound form of the cytoplasmic tail of the Env-glycoprotein: lack of transcomplementation of defective HIV-1 virions encoding C-terminally truncated Env

    OpenAIRE

    Bosch Valerie; Pfeiffer Tanya; Holtkotte Denise

    2006-01-01

    Abstract H9-T-cells do not support the replication of mutant HIV-1 encoding Env protein lacking its long cytoplasmic C-terminal domain (Env-CT). Here we describe the generation of a H9-T-cell population constitutively expressing the HIV-1 Env-CT protein domain anchored in the cellular membrane by it homologous membrane-spanning domain (TMD). We confirmed that the Env-TMD-CT protein was associated with cellular membranes, that its expression did not have any obvious cytotoxic effects on the ce...

  8. Generation of H9 T-cells stably expressing a membrane-bound form of the cytoplasmic tail of the Env-glycoprotein: lack of transcomplementation of defective HIV-1 virions encoding C-terminally truncated Env

    Directory of Open Access Journals (Sweden)

    Bosch Valerie

    2006-05-01

    Full Text Available Abstract H9-T-cells do not support the replication of mutant HIV-1 encoding Env protein lacking its long cytoplasmic C-terminal domain (Env-CT. Here we describe the generation of a H9-T-cell population constitutively expressing the HIV-1 Env-CT protein domain anchored in the cellular membrane by it homologous membrane-spanning domain (TMD. We confirmed that the Env-TMD-CT protein was associated with cellular membranes, that its expression did not have any obvious cytotoxic effects on the cells and that it did not affect wild-type HIV-1 replication. However, as measured in both a single-round assay as well as in spreading infections, replication competence of mutant pNL-Tr712, lacking the Env-CT, was not restored in this H9 T-cell population. This means that the Env-CT per se cannot transcomplement the replication block of HIV-1 virions encoding C-terminally truncated Env proteins and suggests that the Env-CT likely exerts its function only in the context of the complete Env protein.

  9. Dine marker har DNA

    DEFF Research Database (Denmark)

    Eckholdt, Annette; Winding, Anne; Krogh, Paul Henning

    2017-01-01

    Ordet "biodiversitet" og at det er noget, vi skal have mere af, nævnes hyppigt. Men hvad er biodiversitet, og hvordan måles det? Agrologisk har bedt et par eksperter fra Aarhus Universitet forklare, hvordan et DNA-aftryk af jord og vand kan erstatte optællinger i felten og sige noget om biodivers......Ordet "biodiversitet" og at det er noget, vi skal have mere af, nævnes hyppigt. Men hvad er biodiversitet, og hvordan måles det? Agrologisk har bedt et par eksperter fra Aarhus Universitet forklare, hvordan et DNA-aftryk af jord og vand kan erstatte optællinger i felten og sige noget om...

  10. Colony polymerase chain reaction of stably transfected Trypanosoma cruzi grown on solid medium

    Directory of Open Access Journals (Sweden)

    Wagner G dos Santos

    2000-01-01

    Full Text Available Tools for the genetic manipulation of Trypanosoma cruzi are largely unavailable, although several vectors for transfection of epimastigotes and expression of foreign or recombinant genes have been developed. We have previously constructed several plasmid vectors in which recombinant genes are expressed in T. cruzi using the rRNA promoter. In this report, we demonstrate that one of these vectors can simultaneously mediate expression of neomycin phosphotransferase and green fluorescent protein when used to stably transfect cultured epimastigotes. These stably transfected epimastigotes can be selected and cloned as unique colonies on solid medium. We describe a simple colony PCR approach to the screening of these T. cruzi colonies for relevant genes. Thus, the methodologies outlined herein provide important new tools for the genetic dissection of this important parasite.

  11. Persistent human cardiac Na+ currents in stably transfected mammalian cells

    Science.gov (United States)

    Wang, Ging Kuo; Russell, Gabriella; Wang, Sho-Ya

    2013-01-01

    Miniature persistent late Na+ currents in cardiomyocytes have been linked to arrhythmias and sudden death. The goals of this study are to establish a stable cell line expressing robust persistent cardiac Na+ currents and to test Class 1 antiarrhythmic drugs for selective action against resting and open states. After transient transfection of an inactivation-deficient human cardiac Na+ channel clone (hNav1.5-CW with L409C/A410W double mutations), transfected mammalian HEK293 cells were treated with 1 mg/ml G-418. Individual G-418-resistant colonies were isolated using glass cylinders. One colony with high expression of persistent Na+ currents was subjected to a second colony selection. Cells from this colony remained stable in expressing robust peak Na+ currents of 996 ± 173 pA/pF at +50 mV (n = 20). Persistent late Na+ currents in these cells were clearly visible during a 4-second depolarizing pulse albeit decayed slowly. This slow decay is likely due to slow inactivation of Na+ channels and could be largely eliminated by 5 μM batrachotoxin. Peak cardiac hNav1.5-CW Na+ currents were blocked by tetrodotoxin with an IC50 value of 2.27 ± 0.08 μM (n = 6). At clinic relevant concentrations, Class 1 antiarrhythmics are much more selective in blocking persistent late Na+ currents than their peak counterparts, with a selectivity ratio ranging from 80.6 (flecainide) to 3 (disopyramide). We conclude that (1) Class 1 antiarrhythmics differ widely in their resting- vs. open-channel selectivity, and (2) stably transfected HEK293 cells expressing large persistent hNav1.5-CW Na+ currents are suitable for studying as well as screening potent open-channel blockers. PMID:23695971

  12. Harnessing solar heat

    CERN Document Server

    Norton, Brian

    2013-01-01

    Systems engineered by man to harness solar heat in a controlled manner now include a diverse range of technologies each serving distinctive needs in particular climate contexts. This text covers the breadth of solar energy technologies for the conversion of solar energy to provide heat, either as the directly-used output or as an intermediary to other uses such as power generation or cooling. It is a wholly updated, extended and revised version of “Solar Energy Thermal Technology” first published in 1992. The text draws on the own author’s research and that of numerous colleagues and

  13. Evaluation of the Expression of Amyloid Precursor Protein and the Ratio of Secreted Amyloid Beta 42 to Amyloid Beta 40 in SH-SY5Y Cells Stably Transfected with Wild-Type, Single-Mutant and Double-Mutant Forms of the APP Gene for the Study of Alzheimer's Disease Pathology.

    Science.gov (United States)

    Pahrudin Arrozi, Aslina; Shukri, Siti Nur Syazwani; Wan Ngah, Wan Zurinah; Mohd Yusof, Yasmin Anum; Ahmad Damanhuri, Mohd Hanafi; Makpol, Suzana

    2017-11-01

    Neuroblastoma cell lines such as SH-SY5Y are the most frequently utilized models in neurodegenerative research, and their use has advanced the understanding of the pathology of neurodegeneration over the past few decades. In Alzheimer's disease (AD), several pathogenic mutations have been described, all of which cause elevated levels of pathological hallmarks such as amyloid-beta (Aβ). Although the genetics of Alzheimer's disease is well known, familial AD only accounts for a small number of cases in the population, with the rest being sporadic AD, which contains no known mutations. Currently, most of the in vitro models used to study AD pathogenesis only examine the level of Aβ42 as a confirmation of successful model generation and only perform comparisons between wild-type APP and single mutants of the APP gene. Recent findings have shown that the Aβ42/40 ratio in cerebrospinal fluid (CSF) is a better diagnostic indicator for AD patients than is Aβ42 alone and that more extensive Aβ formation, such as accumulation of intraneuronal Aβ, Aβ plaques, soluble oligomeric Aβ (oAβ), and insoluble fibrillar Aβ (fAβ) occurs in TgCRND8 mice expressing a double-mutant form (Swedish and Indiana) of APP, later leading to greater progressive impairment of the brain. In this study, we generated SH-SY5Y cells stably transfected separately with wild-type APP, the Swedish mutation of APP, and the Swedish and Indiana mutations of APP and evaluated the APP expression as well as the Aβ42/40 ratio in those cells. The double-mutant form of APP (Swedish/Indiana) expressed markedly high levels of APP protein and showed a high Aβ2/40 ratio compared to wild-type and single-mutant cells.

  14. HARNESSING BIG DATA VOLUMES

    Directory of Open Access Journals (Sweden)

    Bogdan DINU

    2014-04-01

    Full Text Available Big Data can revolutionize humanity. Hidden within the huge amounts and variety of the data we are creating we may find information, facts, social insights and benchmarks that were once virtually impossible to find or were simply inexistent. Large volumes of data allow organizations to tap in real time the full potential of all the internal or external information they possess. Big data calls for quick decisions and innovative ways to assist customers and the society as a whole. Big data platforms and product portfolio will help customers harness to the full the value of big data volumes. This paper deals with technical and technological issues related to handling big data volumes in the Big Data environment.

  15. Environmental Control Unit Harness Project

    Science.gov (United States)

    Oliva-Buisson, Yvette J.

    2014-01-01

    Testing four new Environmental Control Unit Harnesses for improved user comfort during SCAPE operations. Phase I, testing in a lab environment, Phase II will continue testing the best candidates in a field environment.

  16. Causal boundary for stably causal space-times

    International Nuclear Information System (INIS)

    Racz, I.

    1987-12-01

    The usual boundary constructions for space-times often yield an unsatisfactory boundary set. This problem is reviewed and a new solution is proposed. An explicit identification rule is given on the set of the ideal points of the space-time. This construction leads to a satisfactory boundary point set structure for stably causal space-times. The topological properties of the resulting causal boundary construction are examined. For the stably causal space-times each causal curve has a unique endpoint on the boundary set according to the extended Alexandrov topology. The extension of the space-time through the boundary is discussed. To describe the singularities the defined boundary sets have to be separated into two disjoint sets. (D.Gy.) 8 refs

  17. Harnessing off-target effects

    DEFF Research Database (Denmark)

    Saginc, Gaye; Voellmy, Franziska; Linding, Rune

    2017-01-01

    The 'off-targets' of a drug are often poorly characterized yet could be harnessed in the treatment of complex diseases. A recent study used a small-molecule screening in non-small-cell lung cancer to repurpose an FDA-approved ALK/IGF1R inhibitor and uncover its mechanism of action.......The 'off-targets' of a drug are often poorly characterized yet could be harnessed in the treatment of complex diseases. A recent study used a small-molecule screening in non-small-cell lung cancer to repurpose an FDA-approved ALK/IGF1R inhibitor and uncover its mechanism of action....

  18. Harnessing Aquatic Physicochemical Parameters Influencing ...

    African Journals Online (AJOL)

    The management-oriented background for harnessing aquatic physicochemical parameters influencing macro invertebrate fauna of Anambra River basin for sustainable fish productivity was studied. The intra seasonal variability in the water quality of the river revealed mean transparency of 1.79 cm, Conductivity of 28.81 ...

  19. Harness the Power of Technology

    Science.gov (United States)

    Duncan, Arne

    2011-01-01

    Today, U.S. educators are teaching in the midst of a technological revolution. Technology promises to provide innovative solutions in the nation's classrooms, just as it has transformed the way people communicate, socialize, and conduct business. In this article, the author argues that now is the time to harness technology to revolutionize the way…

  20. Optimal energy growth in a stably stratified shear flow

    Science.gov (United States)

    Jose, Sharath; Roy, Anubhab; Bale, Rahul; Iyer, Krithika; Govindarajan, Rama

    2018-02-01

    Transient growth of perturbations by a linear non-modal evolution is studied here in a stably stratified bounded Couette flow. The density stratification is linear. Classical inviscid stability theory states that a parallel shear flow is stable to exponentially growing disturbances if the Richardson number (Ri) is greater than 1/4 everywhere in the flow. Experiments and numerical simulations at higher Ri show however that algebraically growing disturbances can lead to transient amplification. The complexity of a stably stratified shear flow stems from its ability to combine this transient amplification with propagating internal gravity waves (IGWs). The optimal perturbations associated with maximum energy amplification are numerically obtained at intermediate Reynolds numbers. It is shown that in this wall-bounded flow, the three-dimensional optimal perturbations are oblique, unlike in unstratified flow. A partitioning of energy into kinetic and potential helps in understanding the exchange of energies and how it modifies the transient growth. We show that the apportionment between potential and kinetic energy depends, in an interesting manner, on the Richardson number, and on time, as the transient growth proceeds from an optimal perturbation. The oft-quoted stabilizing role of stratification is also probed in the non-diffusive limit in the context of disturbance energy amplification.

  1. HeLa-LAV, an epithelial cell line stably infected with HIV-1.

    Science.gov (United States)

    Berg, J; Doe, B; Steimer, K S; Wabl, M

    1991-01-01

    An HeLa-LAV cell line was established by infecting and subcloning previously described CD4-expressing HeLa cells with HIV-1. Cells of this line stably synthesize all major HIV proteins, release infectious particles of HIV-1, but do not die even after long term culture. More than 90% of the cells express the envelope protein gp120 on the surface. The cells can be easily and efficiently labeled with 51chromium, and exhibit a low spontaneous release. Because they are susceptible to killing by allogeneic cytotoxic T cells (CTL) when targeted to gp120, they ought to be a useful source of target cells in any kind of HIV-specific killing assays. The cells may also help studies on HIV replication in non-lymphatic/non-monocytic cells. The HeLa-LAV cell line will be freely available from the AIDS Research and Reference Reagent Program.

  2. HARNESSING INNOVATION POTENTIAL OF CROWDSOURCING

    OpenAIRE

    Slavica ROCHESKA; Marjan ANGELESKI; Dimitar NIKOLOSKI; Gjorgji MANCHESKI

    2014-01-01

    The new innovation paradigm is based upon articulation of external and internal sources of innovation. Nowadays, crowdsourcing is coming up as significant external source in innovation processes of the companies. Crowdsourcing enables harnessing of initiatives, ideas, solutions and knowledge of the crowd so as to enhance innovation performance and induce a value creation. Given that existing innovation literature does not cover sufficiently the issues related to crowdsourcing this paper is to...

  3. Materials in Harnessing Solar Power

    Indian Academy of Sciences (India)

    64

    Mission (JNNSM) [8] to boost India's solar energy harnessing capability. The mission document .... Through “The Solar Mission 2020”, Government of India has projected various goals and time-lines to ...... [40] Kim H S, Lee C R, Im J H, Lee K B, Moehl T, Marchioro A, Moon S J, Humphry-Baker. R, Yum J H, Moser J E, ...

  4. Assembly of homotrimeric type XXI minicollagen by coexpression of prolyl 4-hydroxylase in stably transfected Drosophila melanogaster S2 cells.

    Science.gov (United States)

    Li, Hsiu-Chuan; Huang, Chuan-Chuan; Chen, Sung-Fang; Chou, Min-Yuan

    2005-10-21

    We established stably transfected insect cell lines containing cDNAs encoding the alpha and beta subunits of human prolyl 4-hydroxylase in both Trichoplusia ni and Drosophila melanogaster S2 cells. The expression level and enzymatic activity of recombinant prolyl 4-hydroxylase produced in the Drosophila expression system were significantly higher than those produced in the T. ni system. We further characterized the involvement of prolyl 4-hydroxylase in the assembly of the three alpha chains to form trimeric type XXI minicollagen, which comprises the intact C-terminal non-collagenous (NC1) and collagenous domain (COL1), in the Drosophila system. When minicollagen XXI was stably expressed in Drosophila S2 cells alone, negligible amounts of interchain disulfide-bonded trimers were detected in the culture media. However, minicollagen XXI was secreted as disulfide-bonded homotrimers by coexpression with prolyl 4-hydroxylase in the stably transfected Drosophila S2 cells. Minicollagen XXI coexpressed with prolyl 4-hydroxylase contained sufficient amounts of hydroxyproline to form thermal stable pepsin-resistant triple helices consisting of both interchain and non-interchain disulfide-bonded trimers. These results demonstrate that a sufficient amount of active prolyl 4-hydroxylase is required for the assembly of type XXI collagen triple helices in Drosophila cells and the trimeric assembly is governed by the C-terminal collagenous domain.

  5. Stably Doped Conducting Polymer Nanoshells by Surface Initiated Polymerization.

    Science.gov (United States)

    Li, Junwei; Yoon, Soon Joon; Hsieh, Bao-Yu; Tai, Wanyi; O'Donnell, Matthew; Gao, Xiaohu

    2015-12-09

    Despite broad applications ranging from electronics to biomedical sensing and imaging, a long-standing problem of conducting polymers is the poor resistance to dedoping, which directly affects their signature electrical and optical properties. This problem is particularly significant for biomedical uses because of fast leaching of dopant ions in physiological environments. Here, we describe a new approach to engineer multimodal core-shell nanoparticles with a stably doped conductive polymer shell in biological environments. It was achieved by making a densely packed polymer brush rather than changing its molecular structure. Polyaniline (PANI) was used as a model compound due to its concentrated near-infrared (NIR) absorption. It was grafted onto a magnetic nanoparticle via a polydopamine intermediate layer. Remarkably, at pH 7 its conductivity is ca. 2000× higher than conventional PANI nanoshells. Similarly, its NIR absorption is enhanced by 2 orders of magnitude, ideal for photothermal imaging and therapy. Another surprising finding is its nonfouling property, even outperforming polyethylene glycol. This platform technology is also expected to open exciting opportunities in engineering stable conductive materials for electronics, imaging, and sensing.

  6. Dynamics of mixed convective-stably-stratified fluids

    Science.gov (United States)

    Couston, L.-A.; Lecoanet, D.; Favier, B.; Le Bars, M.

    2017-09-01

    We study the dynamical regimes of a density-stratified fluid confined between isothermal no-slip top and bottom boundaries (at temperatures Tt and Tb) via direct numerical simulation. The thermal expansion coefficient of the fluid is temperature dependent and chosen such that the fluid density is maximum at the inversion temperature Tb>Ti>Tt . Thus, the lower layer of the fluid is convectively unstable while the upper layer is stably stratified. We show that the characteristics of the convection change significantly depending on the degree of stratification of the stable layer. For strong stable stratification, the convection zone coincides with the fraction of the fluid that is convectively unstable (i.e., where T >Ti ), and convective motions consist of rising and sinking plumes of large density anomaly, as is the case in canonical Rayleigh-Bénard convection; internal gravity waves are generated by turbulent fluctuations in the convective layer and propagate in the upper layer. For weak stable stratification, we demonstrate that a large fraction of the stable fluid (i.e., with temperature T phenomenological description of the transition between the regimes of plume-dominated and entrainment-dominated convection through analysis of the differences in the heat transfer mechanisms, kinetic energy density spectra, and probability density functions for different stratification strengths. Importantly, we find that the effect of the stable layer on the convection decreases only weakly with increasing stratification strength, meaning that the dynamics of the stable layer and convection should be studied self-consistently in a wide range of applications.

  7. Har du rådne kunder?

    DEFF Research Database (Denmark)

    Grünbaum, Niels Nolsøe

    2015-01-01

    Tætte kunderelationer er ikke altid guld værd. Har du modet til at skille dig af med kunder, som ødelægger din forretning?......Tætte kunderelationer er ikke altid guld værd. Har du modet til at skille dig af med kunder, som ødelægger din forretning?...

  8. Alle har en historie, IV

    DEFF Research Database (Denmark)

    Ydegaard, Torbjørn

    Alle har en historie IV: Pædagogik med Freedom Writers lader et væld af pædagogiske facetter udspringe af den kendte skole-film Freedom Writers’ scener og handlinger. Der er både fokus på en almen tilgang til pædagogik og på Freedom Writer-metodikken, dels gennem de filmscener, der sættes til...... drøftelse, dels gennem foreslåede øvelser. Det styrende princip i bogen er filmens scener og kronologi. Til hver afdeling af filmen knytter der sig oplæg til pædagogiske drøftelser primært rettet mod udsatte unge – og unge i al almindelighed – samt øvelser, der kan anvendes i mange forskellige...... undervisningssammenhænge. Bogens målgruppe er først og fremmest studerende inden for de pædagogiske fagområder, samt professionelle, der arbejder med unge, uanset baggrund. Torbjørn Ydegaard er kandidat i pædagogik, lektor ved University College Syd og uddannet Freedom Writer Teacher....

  9. A TCP-Over-UDP Test Harness

    Energy Technology Data Exchange (ETDEWEB)

    Dunigan, TH

    2002-10-31

    This report describes an implementation of a TCP-like protocol that runs over UDP. This TCP-like implementation, which does not require kernel modifications, provides a test harness for evaluating variations of the TCP transport protocol over the Internet. The test harness provides a tunable and instrumented version of TCP that supports Reno, NewReno, and SACK/FACK. The test harness can also be used to characterize the TCP-performance of a network path. Several case studies illustrate how one can tune the transport protocol to improve performance.

  10. Characterization of cell lines stably transfected with rubella virus replicons

    International Nuclear Information System (INIS)

    Tzeng, Wen-Pin; Xu, Jie; Frey, Teryl K.

    2012-01-01

    Rubella virus (RUBV) replicons expressing a drug resistance gene and a gene of interest were used to select cell lines uniformly harboring the replicon. Replicons expressing GFP and a virus capsid protein GFP fusion (C-GFP) were compared. Vero or BHK cells transfected with either replicon survived drug selection and grew into a monolayer. However, survival was ∼9-fold greater following transfection with the C-GFP-replicon than with the GFP-expressing replicon and while the C-GFP-replicon cells grew similarly to non-transfected cells, the GFP-replicon cells grew slower. Neither was due to the ability of the CP to enhance RNA synthesis but survival during drug selection was correlated with the ability of CP to inhibit apoptosis. Additionally, C-GFP-replicon cells were not cured of the replicon in the absence of drug selection. Interferon-alpha suppressed replicon RNA and protein synthesis, but did not cure the cells, explaining in part the ability of RUBV to establish persistent infections.

  11. Characterization of cell lines stably transfected with rubella virus replicons

    Energy Technology Data Exchange (ETDEWEB)

    Tzeng, Wen-Pin; Xu, Jie [Department of Biology, Georgia State University, P.O. Box 4010, Atlanta GA 30302-4010 (United States); Frey, Teryl K., E-mail: tfrey@gsu.edu [Department of Biology, Georgia State University, P.O. Box 4010, Atlanta GA 30302-4010 (United States)

    2012-07-20

    Rubella virus (RUBV) replicons expressing a drug resistance gene and a gene of interest were used to select cell lines uniformly harboring the replicon. Replicons expressing GFP and a virus capsid protein GFP fusion (C-GFP) were compared. Vero or BHK cells transfected with either replicon survived drug selection and grew into a monolayer. However, survival was {approx}9-fold greater following transfection with the C-GFP-replicon than with the GFP-expressing replicon and while the C-GFP-replicon cells grew similarly to non-transfected cells, the GFP-replicon cells grew slower. Neither was due to the ability of the CP to enhance RNA synthesis but survival during drug selection was correlated with the ability of CP to inhibit apoptosis. Additionally, C-GFP-replicon cells were not cured of the replicon in the absence of drug selection. Interferon-alpha suppressed replicon RNA and protein synthesis, but did not cure the cells, explaining in part the ability of RUBV to establish persistent infections.

  12. Design Process Improvement for Electric CAR Harness

    Science.gov (United States)

    Sawatdee, Thiwarat; Chutima, Parames

    2017-06-01

    In an automobile parts design company, the customer satisfaction is one of the most important factors for product design. Therefore, the company employs all means to focus its product design process based on the various requirements of customers resulting in high number of design changes. The objective of this research is to improve the design process of the electric car harness that effects the production scheduling by using Fault Tree Analysis (FTA) and Failure Mode and Effect Analysis (FMEA) as the main tools. FTA is employed for root cause analysis and FMEA is used to ranking a High Risk Priority Number (RPN) which is shows the priority of factors in the electric car harness that have high impact to the design of the electric car harness. After the implementation, the improvements are realized significantly since the number of design change is reduced from 0.26% to 0.08%.

  13. Stem cells expanded from the human embryonic hindbrain stably retain regional specification and high neurogenic potency.

    Science.gov (United States)

    Tailor, Jignesh; Kittappa, Raja; Leto, Ketty; Gates, Monte; Borel, Melodie; Paulsen, Ole; Spitzer, Sonia; Karadottir, Ragnhildur Thora; Rossi, Ferdinando; Falk, Anna; Smith, Austin

    2013-07-24

    Stem cell lines that faithfully maintain the regional identity and developmental potency of progenitors in the human brain would create new opportunities in developmental neurobiology and provide a resource for generating specialized human neurons. However, to date, neural progenitor cultures derived from the human brain have either been short-lived or exhibit restricted, predominantly glial, differentiation capacity. Pluripotent stem cells are an alternative source, but to ascertain definitively the identity and fidelity of cell types generated solely in vitro is problematic. Here, we show that hindbrain neuroepithelial stem (hbNES) cells can be derived and massively expanded from early human embryos (week 5-7, Carnegie stage 15-17). These cell lines are propagated in adherent culture in the presence of EGF and FGF2 and retain progenitor characteristics, including SOX1 expression, formation of rosette-like structures, and high neurogenic capacity. They generate GABAergic, glutamatergic and, at lower frequency, serotonergic neurons. Importantly, hbNES cells stably maintain hindbrain specification and generate upper rhombic lip derivatives on exposure to bone morphogenetic protein (BMP). When grafted into neonatal rat brain, they show potential for integration into cerebellar development and produce cerebellar granule-like cells, albeit at low frequency. hbNES cells offer a new system to study human cerebellar specification and development and to model diseases of the hindbrain. They also provide a benchmark for the production of similar long-term neuroepithelial-like stem cells (lt-NES) from pluripotent cell lines. To our knowledge, hbNES cells are the first demonstration of highly expandable neuroepithelial stem cells derived from the human embryo without genetic immortalization.

  14. Økonomer har et forklaringsproblem

    DEFF Research Database (Denmark)

    Rangvid, Jesper

    2016-01-01

    Markederne toppede i april/maj 2015 og har haft det svært siden. Meget tyder 2016 er startet med voldsomme fald på de globale aktiemarkeder. Det er muligvis med rette for nogle banker, men overordnet er sektoren mere robust, ikke mindst grundet bedre regulering, end før finanskrisen i 2008...

  15. Kyllinger har effektivt immunforsvar mod herpes

    DEFF Research Database (Denmark)

    Buchmann, Kurt

    2008-01-01

    Forskere ved Københavns Universitet, Fakultet for Biovidenskab har studeret kyllingens MHC molekyler og kan derigennem forklare hvorfor en særlig stamme indenfor denne art er modstandsdygtige mod en særlig herpesvirus. Udgivelsesdato: 18. januar 2008...

  16. Appropriate administrative structures in harnessing water resources ...

    African Journals Online (AJOL)

    Appropriate administrative structures in harnessing water resources for sustainable growth in Nigeria. Lekan Oyebande. Abstract. No Abstract. Journal of Mining and Geology Vol. 42(1) 2006: 21-30. Full Text: EMAIL FULL TEXT EMAIL FULL TEXT · DOWNLOAD FULL TEXT DOWNLOAD FULL TEXT.

  17. Medicinalindustrien har brug for diagnoser som ADHD

    DEFF Research Database (Denmark)

    Fugl, Marie

    2008-01-01

    Det er ikke første gang, at medicinalindustriens evne til at finde nye markeder har medført nye eller bredere diagnoser. I det forrige årti så vi et parallelt forløb mellem et boom i antallet af depressionsdiagnoser og forbruget af lykkepiller. Interview med sociolog Thomas Brante. Udgivelsesdato...

  18. Hvor mange betydninger har et ord?

    DEFF Research Database (Denmark)

    Bergenholtz, Henning; Toftgaard Madsen, Jens August

    2017-01-01

    artikler, som August har skrevet til Den Danske Betydningsordbog: trøske, tonisk, tonnage, topskud, topstykke, toradet, tornerosesøvn, touch, trykkoger, træhammer, trøffel. Ud fra analysen af disse eksempler udleder vi følgende metodiske tese: I forbindelse med receptionsproblem laves polysemi, når et...

  19. Extracellular matrix and hormones transcriptionally regulate bovine. beta. -casein 5 prime sequences in stably transfected mouse mammary cells

    Energy Technology Data Exchange (ETDEWEB)

    Schmidhauser, C. Bissell, M.J. (Univ. of California, Berkeley (United States)); Myers, C.A.; Casperson, G.F. (Monsanto Corporate Research, St. Louis, MO (United States))

    1990-12-01

    Milk protein regulation involves synergistic action of lactogenic hormones and extracellular matrix (ECM). It is well established that substratum has a dramatic effect on morphology and function of mammary cells. The molecular mechanisms that regulate the ECM- and hormone-dependent gene expression, however, have not been resolved. To address this question, a subpopulation (designated CID 9) of the mouse mammary epithelial cell strain COMMA-2D has been developed in which more than 35% of the cells express {beta}-casein, form alveoli-like structures when plated onto a reconstituted basement membrane, and secrete {beta}-casein undirectionally into a lumen. These cells were stably transfected with a series of chloramphenicol acetyltransferase (CAT) fusion genes to study transcriptional regulation of the bovine {beta}-casein gene. The expression of CAT in these lines demonstrated a striking matrix and hormone dependency. This regulation occurered primarily at the transcriptional level and was dependent on the length of the 5{prime} flanking region of the {beta}-casein promotor. Both matrix and hormonal control of transcription occurred within at least the first 1790 base pairs upstream and/or 42 base pairs downstream of the transcriptional initiation site. The ECM effect was independent of glucocorticoid stimulation. However, prolactin was essential and hydrocortisone further increased CAT expression. Endogenous {beta}-casein expression in these lines was similar to that of the parent CID 9 cells. Our data indicate the existence of matrix-dependent elements that regulate transcription.

  20. Dry Electrode Harness System For Wireless 12-LEAD ECG Project

    Data.gov (United States)

    National Aeronautics and Space Administration — The dry-electrode harness is composed of a variety of components.  The harness interfaces with the human via a commercial-off-the-shelf (COTS) FDA cleared dry...

  1. A germline chromothripsis event stably segregating in 11 individuals through three generations

    DEFF Research Database (Denmark)

    Bertelsen, Birgitte; Nazaryan-Petersen, Lusine; Sun, Wei

    2016-01-01

    PURPOSE: Parentally transmitted germ-line chromothripsis (G-CTH) has been identified in only a few cases. Most of these rearrangements were stably transmitted, in an unbalanced form, from a healthy mother to her child with congenital abnormalities probably caused by de novo copy-number changes of...

  2. Targeted surface expression of an exogenous antigen in stably transfected babesia bovis

    Science.gov (United States)

    Babesia bovis is a tick-borne intraerythocytic protozoan responsible for acute disease in cattle which can be controlled by vaccination with attenuated B. bovis strains. Emerging B. bovis transfection technologies may increase the usefulness of these live vaccines. Here we propose using transfected ...

  3. An Archaea 5S rRNA analog is stably expressed in Escherichia coli

    Science.gov (United States)

    Yang, Y.; Fox, G. E.

    1996-01-01

    Mini-genes for 5S-like rRNA were constructed. These genes had a sequence which largely resembles that of the naturally occurring 5S rRNA of a bacterium, Halococcus morrhuae, which phylogenetically belongs to the Archaea. Plasmids carrying the mini-genes were transformed into Escherichia coli (Ec). Ribosomal incorporation was not a prerequisite for stable accumulation of the RNA product. However, only those constructs with a well-base-paired helix I accumulated RNA product. This result strongly implies that this aspect of the structure is likely to be an important condition for stabilizing 5S rRNA-like products. The results are consistent with our current understanding of 5S rRNA processing in Ec. When used in conjunction with rRNA probe technology, the resulting chimeric RNA may be useful as a monitoring tool for genetically engineered microorganisms or naturally occurring organisms that are released into the environment.

  4. Activation of KCNQ5 channels stably expressed in HEK293 cells by BMS-204352

    DEFF Research Database (Denmark)

    Dupuis, Delphine S; Schrøder, Rikke L; Jespersen, Thomas

    2002-01-01

    The novel anti-ischemic compound, BMS-204352 ((3S)-(+)-(5-chloro-2-methoxyphenyl)-1,3-dihydro-3-fluoro-6-(trifluoromethyl)-2H-indol-2-one)), strongly activates the voltage-gated K+ channel KCNQ5 in a concentration-dependent manner with an EC50 of 2.4 microM. At 10 microM, BMS-204352 increased......, BMS-204352 (10 microM) did not significantly shift the KCNQ5 activation curves (threshold and potential for half-activation, V1/2), as observed for the other KCNQ channels. In the presence of BMS-204352, the activation and deactivation kinetics of the KCNQ5 currents were slowed as the slow activation...

  5. Harnessing supramolecular peptide nanotechnology in biomedical applications

    OpenAIRE

    Chan, Kiat Hwa; Lee, Wei Hao; Zhuo, Shuangmu; Ni, Ming

    2017-01-01

    Kiat Hwa Chan,1 Wei Hao Lee,2 Shuangmu Zhuo,3 Ming Ni3 1Division of Science, Yale-NUS College, Singapore; 2Department of Chemistry, Krieger School of Arts & Sciences, Johns Hopkins University, Baltimore, MD, USA; 3Fujian Provincial Key Laboratory for Photonics Technology, Key Laboratory of OptoElectronic Science and Technology for Medicine of Ministry of Education, Fujian Normal University, Fuzhou, People’s Republic of China Abstract: The harnessing of peptides in biomedic...

  6. Harnessing the Microbiome to Enhance Cancer Immunotherapy

    Directory of Open Access Journals (Sweden)

    Michelle H. Nelson

    2015-01-01

    Full Text Available The microbiota plays a key role in regulating the innate and adaptive immune system. Herein, we review the immunological aspects of the microbiota in tumor immunity in mice and man, with a focus on toll-like receptor (TLR agonists, vaccines, checkpoint modulators, chemotherapy, and adoptive T cell transfer (ACT therapies. We propose innovative treatments that may safely harness the microbiota to enhance T cell-based therapies in cancer patients. Finally, we highlight recent developments in tumor immunotherapy, particularly novel ways to modulate the microbiome and memory T cell responses to human malignancies.

  7. Harnessing Dendritic Cells for Tumor Antigen Presentation

    Energy Technology Data Exchange (ETDEWEB)

    Nierkens, Stefan [Department of Tumor Immunology, Nijmegen Centre for Molecular Life Sciences, Radboud University Nijmegen Medical Centre, Geert Grooteplein 28, Nijmegen 6525 GA (Netherlands); Janssen, Edith M., E-mail: edith.janssen@cchmc.org [Division of Molecular Immunology, Cincinnati Children' s Hospital Research Foundation, University of Cincinnati College of Medicine, 3333 Burnet Avenue, Cincinnati, OH 45229 (United States)

    2011-04-26

    Dendritic cells (DC) are professional antigen presenting cells that are crucial for the induction of anti-tumor T cell responses. As a consequence, research has focused on the harnessing of DCs for therapeutic interventions. Although current strategies employing ex vivo-generated and tumor-antigen loaded DCs have been proven feasible, there are still many obstacles to overcome in order to improve clinical trial successes and offset the cost and complexity of customized cell therapy. This review focuses on one of these obstacles and a pivotal step for the priming of tumor-specific CD8{sup +} and CD4{sup +} T cells; the in vitro loading of DCs with tumor antigens.

  8. Harnessing supramolecular peptide nanotechnology in biomedical applications.

    Science.gov (United States)

    Chan, Kiat Hwa; Lee, Wei Hao; Zhuo, Shuangmu; Ni, Ming

    2017-01-01

    The harnessing of peptides in biomedical applications is a recent hot topic. This arises mainly from the general biocompatibility of peptides, as well as from the ease of tunability of peptide structure to engineer desired properties. The ease of progression from laboratory testing to clinical trials is evident from the plethora of examples available. In this review, we compare and contrast how three distinct self-assembled peptide nanostructures possess different functions. We have 1) nanofibrils in biomaterials that can interact with cells, 2) nanoparticles that can traverse the bloodstream to deliver its payload and also be bioimaged, and 3) nanotubes that can serve as cross-membrane conduits and as a template for nanowire formation. Through this review, we aim to illustrate how various peptides, in their various self-assembled nanostructures, possess great promise in a wide range of biomedical applications and what more can be expected.

  9. Reduction of vertical transport in two-dimensional stably stratified forced shear flows

    Science.gov (United States)

    Toqué, Nathalie; Lignières, François; Vincent, Alain

    2006-04-01

    The effect of stable stratification on the vertical transport of passive contaminants in forced, stationary, two-dimensional (2D) and inhomogeneous shear turbulence is investigated numerically. The mean flow consists of several superimposed parallel sheared layers in a stably stratified medium. We find that, as stratification increases, the vertical transport decreases much faster than predicted by mixing length estimates. For the highest stratification, particles vertical dispersion nearly vanishes. The proposed interpretation emphasizes the role of weakly sheared layers where the relative increase of the mean horizontal velocity with respect to the root-mean-square (rms) vertical velocity causes the decrease of the Lagrangian correlation timescale.

  10. A KBE Application for Automatic Aircraft Wire Harness Routing

    NARCIS (Netherlands)

    Zhu, Z.; Van Tooren, M.J.L.; La Rocca, G.

    2012-01-01

    Wire harness design is an increasingly complex task. Knowledge Based Engineering (KBE) and optimization techniques can be used to support designers in handling this complexity. The wire harness design process can be divided in three main parts, namely electrical design, configuration design and

  11. 46 CFR 197.324 - Diver's safety harness.

    Science.gov (United States)

    2010-10-01

    ... 46 Shipping 7 2010-10-01 2010-10-01 false Diver's safety harness. 197.324 Section 197.324 Shipping COAST GUARD, DEPARTMENT OF HOMELAND SECURITY (CONTINUED) MARINE OCCUPATIONAL SAFETY AND HEALTH STANDARDS GENERAL PROVISIONS Commercial Diving Operations Equipment § 197.324 Diver's safety harness. Each safety...

  12. Registration of two confection sunflower germplasm Lines, HA-R10 and HA-R11, Resistant to sunflower rust

    Science.gov (United States)

    Two confection sunflower (Helianthus annuus L.) germplasm lines, HA-R10 (Reg. No.xxx, PI670043) and HA-R11 (Reg. No.xxx, PI670044) were developed by the USDA-ARS Sunflower and Plant Biology Research Unit in collaboration with the North Dakota Agricultural Experiment Station and released December, 20...

  13. Registration of two double rust resistant germplasms, HA-R12 and HA-R13 for confection sunflower

    Science.gov (United States)

    The confection sunflower (Helianthus annuus L.) germplasms HA-R12 (Reg. No. ______, PI 673104) and HA-R13 (Reg. No. ______, PI 673105) were developed by the USDA-ARS, Sunflower and Plant Biology Research Unit in collaboration with the North Dakota Agricultural Experiment Station, and released in Jul...

  14. Harnessing adolescent values to motivate healthier eating.

    Science.gov (United States)

    Bryan, Christopher J; Yeager, David S; Hinojosa, Cintia P; Chabot, Aimee; Bergen, Holly; Kawamura, Mari; Steubing, Fred

    2016-09-27

    What can be done to reduce unhealthy eating among adolescents? It was hypothesized that aligning healthy eating with important and widely shared adolescent values would produce the needed motivation. A double-blind, randomized, placebo-controlled experiment with eighth graders (total n = 536) evaluated the impact of a treatment that framed healthy eating as consistent with the adolescent values of autonomy from adult control and the pursuit of social justice. Healthy eating was suggested as a way to take a stand against manipulative and unfair practices of the food industry, such as engineering junk food to make it addictive and marketing it to young children. Compared with traditional health education materials or to a non-food-related control, this treatment led eighth graders to see healthy eating as more autonomy-assertive and social justice-oriented behavior and to forgo sugary snacks and drinks in favor of healthier options a day later in an unrelated context. Public health interventions for adolescents may be more effective when they harness the motivational power of that group's existing strongly held values.

  15. Harnessing supramolecular peptide nanotechnology in biomedical applications

    Directory of Open Access Journals (Sweden)

    Chan KH

    2017-02-01

    Full Text Available Kiat Hwa Chan,1 Wei Hao Lee,2 Shuangmu Zhuo,3 Ming Ni3 1Division of Science, Yale-NUS College, Singapore; 2Department of Chemistry, Krieger School of Arts & Sciences, Johns Hopkins University, Baltimore, MD, USA; 3Fujian Provincial Key Laboratory for Photonics Technology, Key Laboratory of OptoElectronic Science and Technology for Medicine of Ministry of Education, Fujian Normal University, Fuzhou, People’s Republic of China Abstract: The harnessing of peptides in biomedical applications is a recent hot topic. This arises mainly from the general biocompatibility of peptides, as well as from the ease of tunability of peptide structure to engineer desired properties. The ease of progression from laboratory testing to clinical trials is evident from the plethora of examples available. In this review, we compare and contrast how three distinct self-assembled peptide nanostructures possess different functions. We have 1 nanofibrils in biomaterials that can interact with cells, 2 nanoparticles that can traverse the bloodstream to deliver its payload and also be bioimaged, and 3 nanotubes that can serve as cross-membrane conduits and as a template for nanowire formation. Through this review, we aim to illustrate how various peptides, in their various self-assembled nanostructures, possess great promise in a wide range of biomedical applications and what more can be expected. Keywords: peptides, self-assembly, nanotechnology

  16. Wearing a safety harness during treadmill walking influences lower extremity kinematics mainly through changes in ankle regularity and local stability

    Directory of Open Access Journals (Sweden)

    Decker Leslie M

    2012-02-01

    Full Text Available Abstract Background Wearing a harness during treadmill walking ensures the subject's safety and is common practice in biomedical engineering research. However, the extent to which such practice influences gait is unknown. This study investigated harness-related changes in gait patterns, as evaluated from lower extremity kinematics during treadmill walking. Findings Healthy subjects (n = 10 walked on a treadmill at their preferred speed for 3 minutes with and without wearing a harness (LiteGait®, Mobility Research, Inc.. In the former condition, no weight support was provided to the subjects. Lower extremity kinematics was assessed in the sagittal plane from the mean (meanRoM, standard deviation (SDRoM and coefficient of variation (CoVRoM of the hip, knee, and ankle ranges of motion (RoM, as well as from the sample entropy (SampEn and the largest Lyapunov exponent (LyE of the joints' angles. Wearing the harness increased the meanRoM of the hip, the SDRoM and the CoVRoM of the knee, and the SampEn and the LyE of the ankle. In particular, the harness effect sizes for both the SampEn and the LyE of the ankle were large, likely reflecting a meaningful decline in the neuromuscular stabilizing control of this joint. Conclusions Wearing a harness during treadmill walking marginally influences lower extremity kinematics, resulting in more or less subtle changes in certain kinematic variables. However, in cases where differences in gait patterns would be expressed through modifications in these variables, having subjects walk with a harness may mask or reinforce such differences.

  17. Turbulent Mechanical Energy Budget in Stably Stratified Baroclinic Flows over Sloping Terrain

    Science.gov (United States)

    Łobocki, Lech

    2017-09-01

    Analysis of second-moment budget equations in a slope-oriented coordinate frame exhibits the pathways of exchange between the potential energy of mean flow and the total turbulent mechanical energy. It is shown that this process is controlled by the inclination of the potential temperature gradient. Hence, this parameter should be considered in studies of turbulence in slope flows as well as the slope inclination. The concept of turbulent potential energy is generalized to include baroclinicity, and is used to explain the role of along-slope turbulent heat flux in energy conversions. A generalization of static stability criteria for baroclinic conditions is also proposed. In addition, the presence of feedback between the turbulent heat flux and the temperature variance in stably-stratified flows is identified, which implies the existence of oscillatory modes characterized by the Brunt-Väisäla frequency.

  18. Numerical Simulations of Stably Stratified Fluid Flow Using Compact Finite-Difference Schemes

    Science.gov (United States)

    Bodnár, T.; Fraunié, Ph.; Kozel, K.

    2010-09-01

    The aim of this paper is to present the class of high order compact schemes in the context of numerical simulation of stratified flow. The numerical schemes presented here are based on the approach outlined in Lele [1]. The numerical model presented in this contribution is based on the solution of the Boussinesq approximation by a finite-difference scheme. The numerical scheme itself follows the principle of semi-discretization, with high order compact discretization in space, while the time integration is carried out by suitable Runge-Kutta time-stepping scheme. In the case presented here the steady flow was considered and thus the artificial compressibility method was used to resolve the pressure from the modified continuity equation. The test case used to demonstrate the capabilities of the selected model consists of the flow of stably stratified fluid over low, smooth hill.

  19. Harnessing Earth Observations for Disaster Application Science

    Science.gov (United States)

    Green, D. S.

    2015-12-01

    Earth observations have made substantive contributions to the understanding of natural hazards, answering key science questions on the mechanisms, processes and dynamics of changes in the land, air and water. This has been achieved through the ability to advance models and interpret the results through maps and assessments. Disaster application science is focused on the two-way flow of data and information between hazard understanding and the knowledge required for disaster response, relief and recovery. This presentation will examine the integration of results from mature science and technology development in areas including optical imagery, synthetic-aperture radar and geodetic sensors, which together provide new levels of situational awareness. Specific examples will be highlighted from the recent Nepal "Gorkha" earthquake. Optical imagery from a host of satellite missions was used to create a comprehensive mosaic across the region, which when analyzed by a global network of volunteer scientists yielded insight into the extent of induced hazards and impacts. In some cases unique day/night band images provided guidance on areas where energy-dependent infrastructure of livelihoods were disrupted. Earthquake modeling and historical trend analysis revealed areas of potential vulnerability and combined with aftershock analysis to guide areas for urgent analysis and action. The combination of SAR and GPS data, innovative integration and processing approaches and nontraditional data integration approaches resulted in damage proxy maps or where combination with airborne photography, field sightings and crowd sourced reports to assess susceptibility to induced hazards (floods and landslides). Opportunities and challenges to build the science and community relationships, harness the earth observations from multiple agencies and institutions and co-develop timely applications to users will be areas for ongoing collaboration and study.Earth observations have made

  20. Generation and preclinical immunogenicity study of dengue type 2 virus-like particles derived from stably transfected mosquito cells.

    Science.gov (United States)

    Suphatrakul, Amporn; Yasanga, Thippawan; Keelapang, Poonsook; Sriburi, Rungtawan; Roytrakul, Thaneeya; Pulmanausahakul, Rojjanaporn; Utaipat, Utaiwan; Kawilapan, Yanee; Puttikhunt, Chunya; Kasinrerk, Watchara; Yoksan, Sutee; Auewarakul, Prasert; Malasit, Prida; Charoensri, Nicha; Sittisombut, Nopporn

    2015-10-13

    Recent phase IIb/III trials of a tetravalent live attenuated vaccine candidate revealed a need for improvement in the stimulation of protective immunity against diseases caused by dengue type 2 virus (DENV-2). Our attempts to develop particulate antigens for possibly supplementing live attenuated virus preparation involve generation and purification of recombinant DENV-2 virus-like particles (VLPs) derived from stably (prM+E)-expressing mosquito cells. Two VLP preparations generated with either negligible or enhanced prM cleavage exhibited different proportions of spherical particles and tubular particles of variable lengths. In BALB/c mice, VLPs were moderately immunogenic, requiring adjuvants for the induction of strong virus neutralizing antibody responses. VLPs with enhanced prM cleavage induced higher levels of neutralizing antibody than those without, but the stimulatory activity of both VLPs was similar in the presence of adjuvants. Comparison of EDIII-binding antibodies in mice following two adjuvanted doses of these VLPs revealed subtle differences in the stimulation of anti-EDIII binding antibodies. In cynomolgus macaques, VLPs with enhanced prM cleavage augmented strongly neutralizing antibody and EDIII-binding antibody responses in live attenuated virus-primed recipients, suggesting that these DENV-2 VLPs may be useful as the boosting antigen in prime-boost immunization. As the levels of neutralizing antibody induced in macaques with the prime-boost immunization were comparable to those infected with wild type virus, this virus-prime VLP-boost regimen may provide an immunization platform in which a need for robust neutralizing antibody response in the protection against DENV-2-associated illnesses could be tested. Copyright © 2015 Elsevier Ltd. All rights reserved.

  1. Establishment of a human cell line stably overexpressing mouse Nip45 and characterization of Nip45 subcellular localization

    International Nuclear Information System (INIS)

    Hashiguchi, Kohtaro; Ozaki, Masumi; Kuraoka, Isao; Saitoh, Hisato

    2013-01-01

    Highlights: ► A human cell line expressing a mouse Nip45 has facilitated Nip45 analysis. ► Nip45 does not effectively inhibit polySUMOylation in vivo. ► Nip45 interacts directly with SUMO and SUMO chains. ► Nip45 accumulates at PML bodies in response to proteasome inhibition. -- Abstract: The nuclear factor of activated T cells, cytoplasmic, calcineurin dependent 2 interacting protein, Nfatc2ip (Nip45), has been implicated as a crucial coordinator of the immune response and of cellular differentiation in humans and mice, and contains SUMO-like domains in its C-terminal region. However, the significance of its N-terminal region and its correlation to the SUMO modification pathway remain largely uncharacterized. In this study, a human cultured cell line was established, in which FLAG-tagged mouse Nip45 (FLAG-mNip45) was stably overexpressed. Under standard, non-stressful conditions, we detected FLAG-mNip45 diffusely distributed in the nucleus. Intriguingly, proteasome inhibition by MG132 caused FLAG-mNip45, together with SUMOylated proteins, to localize in nuclear domains associated with promyelocytic leukemia protein. Finally, using an in vitro binding assay, we showed interaction of the N-terminal region of mNip45 with both free SUMO-3 and SUMO-3 chains, indicating that Nip45 may, in part, exert its function via interaction with SUMO/SUMOylated proteins. Taken together, our study provides novel information on a poorly characterized mammalian protein and suggests that our newly established cell line will be useful for elucidating the physiological role of Nip45.

  2. Developing a TTCN-3 Test Harness for Legacy Software

    DEFF Research Database (Denmark)

    Okika, Joseph C.; Ravn, Anders Peter; Siddalingaiah, Lokesh

    2006-01-01

    challenge in developing the test harness is to interface a generic test driver to the legacy software and provide a suitable interface for test engineers. The main contribution of this paper is a demonstration of a suitable design for such a test harness. It includes: a TTCN-3 test driver in C++, the legacy...... control software in C, a Graphical User Interface (GUI) and the connectors in Java. Our experience shows that it is feasible to use TTCN-3 in developing a test harness for a legacy software for an embedded system, even when it involves different heterogeneous components.......We describe a prototype test harness for an embedded system which is the control software for a modern marine diesel engine. The operations of such control software requires complete certification. We adopt Testing and Test Control Notation (TTCN-3) to define test cases for this purpose. The main...

  3. Developing a TTCN-3 Test Harness for Legacy Software

    DEFF Research Database (Denmark)

    Okika, Joseph C.; Ravn, Anders Peter; Siddalingaiah, Lokesh

    2006-01-01

    We describe a prototype test harness for an embedded system which is the control software for a modern marine diesel engine. The operations of such control software requires complete certification. We adopt Testing and Test Control Notation (TTCN-3) to define test cases for this purpose. The main...... challenge in developing the test harness is to interface a generic test driver to the legacy software and provide a suitable interface for test engineers. The main contribution of this paper is a demonstration of a suitable design for such a test harness. It includes: a TTCN-3 test driver in C++, the legacy...... control software in C, a Graphical User Interface (GUI) and the connectors in Java. Our experience shows that it is feasible to use TTCN-3 in developing a test harness for a legacy software for an embedded system, even when it involves different heterogeneous components....

  4. Harnessing the power of civil registration and vital statistics systems ...

    International Development Research Centre (IDRC) Digital Library (Canada)

    2018-02-27

    :45pm. This conference will identify how best to harness the global momentum for strengthening civil registration and vital statistics (CRVS) systems to meet the 2030 Sustainable Development Agenda. Policy and practice ...

  5. Har læreplanen (stadig) minimal betydning?

    DEFF Research Database (Denmark)

    Poulsen, Jens Aage

    2016-01-01

    Undervisningsministeriets læreplaner for skolens fag er centrale styredokumenter, der fastlægger formål og rammesætter indhold og form for undervisningen. Men hvilken betydning har de i praksis?...

  6. Modeling Cable-Harness Effects on Spacecraft Structures

    Data.gov (United States)

    National Aeronautics and Space Administration — The proposed research involves modeling the effects of cable-harnesses and wiring on the dynamics of spacecraft structures. As space structure mass decreases due to...

  7. 42 CFR 84.1138 - Head harnesses; minimum requirements.

    Science.gov (United States)

    2010-10-01

    ..., and Mist; Pesticide; Paint Spray; Powered Air-Purifying High Efficiency Respirators and Combination Gas Masks § 84.1138 Head harnesses; minimum requirements. (a) All facepieces shall be equipped with...

  8. Army Social Media: Harnessing the Power of Networked Communications

    Science.gov (United States)

    2011-09-01

    9/1/2011 Army Social Media : harnessing the power of networked communications Report Documentation Page Form ApprovedOMB No. 0704-0188 Public...4. TITLE AND SUBTITLE Army Social Media : harnessing the power of networked communications 5a. CONTRACT NUMBER 5b. GRANT NUMBER 5c. PROGRAM...the Chief of Public Affairs,Online and Social Media Division,1500 Pentagon,Washington,DC,20301 8. PERFORMING ORGANIZATION REPORT NUMBER 9

  9. Edge and core dynamics in harness

    International Nuclear Information System (INIS)

    Ball, R.

    2007-01-01

    Resistive kink oscillations in tokamak plasmas are usually treated as core localized events, yet there there are several mechanisms by which they may interact with the edge dynamics. This suggests that we may regulate edge oscillatory behaviour, or ELMs, by harnessing the natural or contrived sawtooth period and amplitude. In this work I investigate core-edge oscillatory entrainment through direct propagation of heat pulses, inductive coupling, and global higher order resonance effects. In the core of auxiliary heated tokamak plasmas the ineluctable rhythm of slow buildup and rapid conversion of potential energy governs electron and heat radial transport. The growth phase of the sawtooth is accompanied by significant reconnection, then during the collapse the temperature and density in the core fall dramatically. There is evidence from experiments in reversed field pinch devices that ensuing energy fluxes can affect flow shear and confinement at the edge. The basis for this study is the dynamical (BDS) model for edge plasma behavior that was derived from electrostatic resistive MHD equations. The BDS model reflects the major qualitative features of edge dynamics that have been observed, such as L-H transitions and associated ELMs, hysteresis, and spontaneous reversal of poloidal shear flow. Under poorly dissipative conditions the transient behavior of the model can exhibit period-doubling, blue-sky, homoclinic, and other exotic bifurcations. Thus we might ask questions such as: Is it possible to mode-lock the edge dynamics to the core sawteeth? Can we induce, or prevent, a change in direction of shear flow? What about MHD effects? Is core-edge communication one way or is there some feedback? In the simplest prototype for coupled core-edge dynamics I model the sawtooth crash as a periodic power input to the edge potential energy reservoir. This is effected by coupling the BDS model to the dynamical system u = u(1 - u 2 - x 2 ) - ω s x, x = x(1-u 2 -x 2 ) + ω s u

  10. A Case Study of Offshore Advection of Boundary Layer Rolls over a Stably Stratified Sea Surface

    Directory of Open Access Journals (Sweden)

    Nina Svensson

    2017-01-01

    Full Text Available Streaky structures of narrow (8-9 km high wind belts have been observed from SAR images above the Baltic Sea during stably stratified conditions with offshore winds from the southern parts of Sweden. Case studies using the WRF model and in situ aircraft observations indicate that the streaks originate from boundary layer rolls generated over the convective air above Swedish mainland, also supported by visual satellite images showing the typical signature cloud streets. The simulations indicate that the rolls are advected and maintained at least 30–80 km off the coast, in agreement with the streaks observed by the SAR images. During evening when the convective conditions over land diminish, the streaky structures over the sea are still seen in the horizontal wind field; however, the vertical component is close to zero. Thus advected feature from a land surface can affect the wind field considerably for long times and over large areas in coastal regions. Although boundary layer rolls are a well-studied feature, no previous study has presented results concerning their persistence during situations with advection to a strongly stratified boundary layer. Such conditions are commonly encountered during spring in coastal regions at high latitudes.

  11. Silicon homo-heterojunction solar cells: A promising candidate to realize high performance more stably

    Science.gov (United States)

    Tan, Miao; Zhong, Sihua; Wang, Wenjie; Shen, Wenzhong

    2017-08-01

    We have investigated the influences of diverse physical parameters on the performances of a silicon homo-heterojunction (H-H) solar cell, which encompasses both homojunction and heterojunction, together with their underlying mechanisms by the aid of AFORS-HET simulation. It is found that the performances of H-H solar cell are less sensitive to (i) the work function of the transparent conductive oxide layer, (ii) the interfacial density of states at the front hydrogenated amorphous silicon/crystalline silicon (a-Si:H/c-Si) interface, (iii) the peak dangling bond defect densities within the p-type a-Si:H (p-a-Si:H) layer, and (iv) the doping concentration of the p-a-Si:H layer, when compared to that of the conventional heterojunction with intrinsic thin layer (HIT) counterparts. These advantages are due to the fact that the interfacial recombination and the recombination within the a-Si:H region are less affected by all the above parameters, which fundamentally benefit from the field-effect passivation of the homojunction. Therefore, the design of H-H structure can provide an opportunity to produce high-efficiency solar cells more stably.

  12. Silicon homo-heterojunction solar cells: A promising candidate to realize high performance more stably

    Directory of Open Access Journals (Sweden)

    Miao Tan

    2017-08-01

    Full Text Available We have investigated the influences of diverse physical parameters on the performances of a silicon homo-heterojunction (H-H solar cell, which encompasses both homojunction and heterojunction, together with their underlying mechanisms by the aid of AFORS-HET simulation. It is found that the performances of H-H solar cell are less sensitive to (i the work function of the transparent conductive oxide layer, (ii the interfacial density of states at the front hydrogenated amorphous silicon/crystalline silicon (a-Si:H/c-Si interface, (iii the peak dangling bond defect densities within the p-type a-Si:H (p-a-Si:H layer, and (iv the doping concentration of the p-a-Si:H layer, when compared to that of the conventional heterojunction with intrinsic thin layer (HIT counterparts. These advantages are due to the fact that the interfacial recombination and the recombination within the a-Si:H region are less affected by all the above parameters, which fundamentally benefit from the field-effect passivation of the homojunction. Therefore, the design of H-H structure can provide an opportunity to produce high-efficiency solar cells more stably.

  13. The turbulent decay of trailing vortex pairs in stably stratified environments

    Energy Technology Data Exchange (ETDEWEB)

    Holzaepfel, F.; Gerz, T.; Baumann, R.

    2000-03-01

    The decay of trailing vortex pairs in thermally stably stratified environments is investigated by means of large eddy simulations. Results of in-situ measurements in the wakes of different aircraft are used to find appropriate intitializations for the simulation of wake turbulence in the quiescent atmosphere. Furthermore, cases with weak atmospheric turbulence are investigated. It is shown that the early development of the vortices is not affected by turbulence and develops almost identically as in 2D simulations. In a quiescent atmosphere the subsequent vortex decay is controlled by the interaction of short-wave disturbances, owing to the aircraft induced turbulence, and baroclinic vorticity, owing to stable stratification. As a consequence, vertical vorticity streaks between the vortices are induced which are substantially intensified by vortex stretching and finally lead to rapid turbulent wake-vortex decay. When in addition also atmospheric turbulence is present, the long-wave instability is dominantly promoted. For very strong stratification (Fr < 1) it is observed that wake vortices may rebound but lose most of their strength before reaching the flight level. Finally, the simulation results are compared to the predictive capabilities of Greene's approximate model. (orig.)

  14. Harnessing transcription for bioproduction in cyanobacteria

    DEFF Research Database (Denmark)

    Stensjö, Karin; Vavitsas, Konstantinos; Tyystjärvi, Taina

    2018-01-01

    Sustainable production of biofuels and other valuable compounds is one of our future challenges. One tempting possibility is to use photosynthetic cyanobacteria as production factories. Currently, tools for genetic engineering of cyanobacteria are yet not good enough to exploit the full potential...... of cyanobacteria. A wide variety of expression systems will be required to adjust both the expression of heterologous enzyme(s) and metabolic routes to the best possible balance, allowing the optimal production of a particular substance. In bacteria, transcription, especially the initiation of transcription, has...

  15. Advances and limitations of visual conditioning protocols in harnessed bees.

    Science.gov (United States)

    Avarguès-Weber, Aurore; Mota, Theo

    2016-10-01

    Bees are excellent invertebrate models for studying visual learning and memory mechanisms, because of their sophisticated visual system and impressive cognitive capacities associated with a relatively simple brain. Visual learning in free-flying bees has been traditionally studied using an operant conditioning paradigm. This well-established protocol, however, can hardly be combined with invasive procedures for studying the neurobiological basis of visual learning. Different efforts have been made to develop protocols in which harnessed honey bees could associate visual cues with reinforcement, though learning performances remain poorer than those obtained with free-flying animals. Especially in the last decade, the intention of improving visual learning performances of harnessed bees led many authors to adopt distinct visual conditioning protocols, altering parameters like harnessing method, nature and duration of visual stimulation, number of trials, inter-trial intervals, among others. As a result, the literature provides data hardly comparable and sometimes contradictory. In the present review, we provide an extensive analysis of the literature available on visual conditioning of harnessed bees, with special emphasis on the comparison of diverse conditioning parameters adopted by different authors. Together with this comparative overview, we discuss how these diverse conditioning parameters could modulate visual learning performances of harnessed bees. Copyright © 2016 Elsevier Ltd. All rights reserved.

  16. Turbulent jet erosion of a stably stratified gas layer in a nuclear reactor test containment

    Energy Technology Data Exchange (ETDEWEB)

    Ishay, Liel [Department of Mechanical Engineering, Ben-Gurion University of the Negev, Beer-Sheva 84105 (Israel); Bieder, Ulrich [Commissariat à l’énergie atomique et aux énergies alternatives, Centre de SACLAY DEN/SAC/DANS/DM2S/STMF/LMSF, F-91191 Gif-sur-Yvette (France); Ziskind, Gennady [Department of Mechanical Engineering, Ben-Gurion University of the Negev, Beer-Sheva 84105 (Israel); Rashkovan, Alex, E-mail: rashbgu@gmail.com [Physics Department, Nuclear Research Center Negev (NRCN), PO Box 9001, Beer-Sheva 84190 (Israel)

    2015-10-15

    Highlights: • We model stably stratified layer erosion by vertical turbulent round jet. • Separate effect studies are performed as a platform for choosing modeling approach. • A test performed in MISTRA facility, CEA, Saclay is modeled using Fluent and Trio-U codes. • The proposed modeling approach showed good agreement with the MISTRA facility LOWMA-3 test. - Abstract: A number of integral and separate effect experiments were performed in the last two decades for validation of containment computational tools. The main goal of these benchmark experiments was to assess the ability of turbulence models and computational fluid dynamics codes to predict hydrogen concentration distribution and steam condensation rate in a nuclear reactor containment in the course of severe accidents. It appears from the published literature that the predictive capability of the existing computational tools still needs to be improved. This work examines numerically the temporal evolution of helium concentration in the experiment called LOWMA-3, performed in the MISTRA facility of CEA-Saclay, France. In the experiment, helium is used to mimic hydrogen of a real-case accident. The aim of this separate effect experiment, where steam condensation was not involved, is to predict helium concentration field. The conditions of the experiment are such that both the momentum transport and molecular diffusion contributions to the mixing process are of the same order of magnitude (Fr ∼ 1). A commercial CFD code, Fluent, and a CEA in-house code, Trio-U, are used for flow and helium concentration fields temporal evolution prediction in the present study. The preliminary separate effect studies provide guidance to an optimal modeling approach for the LOWMA-3 experiment. Temporal evolution of helium concentration in the stratification layer is shown, and a comparison to the experiment is discussed. It is shown that correct modeling of the round jet flowfield is essential for a reliable

  17. Direct numerical simulations of stably-stratified sheared turbulence: Implications for oceanic mixing

    Science.gov (United States)

    Itsweire, E. C.; Holt, S. E.; Koseff, J. R.; Ferziger, J. H.

    1990-01-01

    Direct numerical simulations of the time evolution of homogeneous stably stratified turbulent shear flows have been performed for several Richardson numbers Ri and Reynolds numbers R(sub lambda) in earlier works. The results show excellent agreement with length scale models developed from laboratory experiments to characterize oceanic turbulence. When the Richardson number Ri is less than the stationary value Ri(sub s), the turbulence intensity grows at all scales, and the growth rate appears to be a function of Ri. The size of the vertical density inversions also increases. On the other hand, when Ri is greater than or equal to Ri(sub s) the largest turbulent eddies become vertically constrained by buoyancy when the Ellison (turbulence) scale L(sub E) and the Ozmidov (buoyancy) scale L(sub O) are equal. At this point, the mixing efficiency is maximal and corresponds to a flux Richardson number R(sub f) = 0.20. The vertical mass flux becomes counter-gradient when epsilon = 19(nu)N(exp 2) and vertical density overturns are suppressed in less than half a Brunt-Vaisala period. The results of the simulations were also recast in terms of the Hydrodynamic Phase Diagram introduced in fossil turbulence models. The so-called point of fossilization occurs when epsilon = 4DCN(exp 2); Gibson proposed 13DCN(exp 2). This value is in agreement with indirect laboratory observations and field observations. Finally, the validity of the steady-state models to estimate vertical eddy diffusivities in the oceanic thermocline is discussed.

  18. Turbulent jet erosion of a stably stratified gas layer in a nuclear reactor test containment

    International Nuclear Information System (INIS)

    Ishay, Liel; Bieder, Ulrich; Ziskind, Gennady; Rashkovan, Alex

    2015-01-01

    Highlights: • We model stably stratified layer erosion by vertical turbulent round jet. • Separate effect studies are performed as a platform for choosing modeling approach. • A test performed in MISTRA facility, CEA, Saclay is modeled using Fluent and Trio-U codes. • The proposed modeling approach showed good agreement with the MISTRA facility LOWMA-3 test. - Abstract: A number of integral and separate effect experiments were performed in the last two decades for validation of containment computational tools. The main goal of these benchmark experiments was to assess the ability of turbulence models and computational fluid dynamics codes to predict hydrogen concentration distribution and steam condensation rate in a nuclear reactor containment in the course of severe accidents. It appears from the published literature that the predictive capability of the existing computational tools still needs to be improved. This work examines numerically the temporal evolution of helium concentration in the experiment called LOWMA-3, performed in the MISTRA facility of CEA-Saclay, France. In the experiment, helium is used to mimic hydrogen of a real-case accident. The aim of this separate effect experiment, where steam condensation was not involved, is to predict helium concentration field. The conditions of the experiment are such that both the momentum transport and molecular diffusion contributions to the mixing process are of the same order of magnitude (Fr ∼ 1). A commercial CFD code, Fluent, and a CEA in-house code, Trio-U, are used for flow and helium concentration fields temporal evolution prediction in the present study. The preliminary separate effect studies provide guidance to an optimal modeling approach for the LOWMA-3 experiment. Temporal evolution of helium concentration in the stratification layer is shown, and a comparison to the experiment is discussed. It is shown that correct modeling of the round jet flowfield is essential for a reliable

  19. Phenomenology of two-dimensional stably stratified turbulence under large-scale forcing

    KAUST Repository

    Kumar, Abhishek

    2017-01-11

    In this paper, we characterise the scaling of energy spectra, and the interscale transfer of energy and enstrophy, for strongly, moderately and weakly stably stratified two-dimensional (2D) turbulence, restricted in a vertical plane, under large-scale random forcing. In the strongly stratified case, a large-scale vertically sheared horizontal flow (VSHF) coexists with small scale turbulence. The VSHF consists of internal gravity waves and the turbulent flow has a kinetic energy (KE) spectrum that follows an approximate k−3 scaling with zero KE flux and a robust positive enstrophy flux. The spectrum of the turbulent potential energy (PE) also approximately follows a k−3 power-law and its flux is directed to small scales. For moderate stratification, there is no VSHF and the KE of the turbulent flow exhibits Bolgiano–Obukhov scaling that transitions from a shallow k−11/5 form at large scales, to a steeper approximate k−3 scaling at small scales. The entire range of scales shows a strong forward enstrophy flux, and interestingly, large (small) scales show an inverse (forward) KE flux. The PE flux in this regime is directed to small scales, and the PE spectrum is characterised by an approximate k−1.64 scaling. Finally, for weak stratification, KE is transferred upscale and its spectrum closely follows a k−2.5 scaling, while PE exhibits a forward transfer and its spectrum shows an approximate k−1.6 power-law. For all stratification strengths, the total energy always flows from large to small scales and almost all the spectral indicies are well explained by accounting for the scale-dependent nature of the corresponding flux.

  20. ”Eldre har ikke slike behov” Eldre og seksualitet

    OpenAIRE

    Paulsen, Kine; Løkken, Stine

    2016-01-01

    Bacheloroppgave sykepleie, 2016 Tittel ”Eldre har ikke slike behov” : Eldre og seksualitet. ”The elderly do not have those needs” : Sexuality in later life. Formål Vi ønsker med denne bacheloroppgaven å belyse dette høyst aktuelle, men tabubelagte temaet. Gjennom erfaringer fra praksis fremstår seksualitet som et tema sykepleiere unnviker å kommunisere om. Oppgaven har som formål å innhente økt kunnskap om eldre og seksualitet, og om hvordan sykepleiere kan ivareta dette beh...

  1. GnRH receptor activation competes at a low level with growth signaling in stably transfected human breast cell lines

    International Nuclear Information System (INIS)

    Morgan, Kevin; Meyer, Colette; Miller, Nicola; Sims, Andrew H; Cagnan, Ilgin; Faratian, Dana; Harrison, David J; Millar, Robert P; Langdon, Simon P

    2011-01-01

    Gonadotrophin releasing hormone (GnRH) analogs lower estrogen levels in pre-menopausal breast cancer patients. GnRH receptor (GnRH-R) activation also directly inhibits the growth of certain cells. The applicability of GnRH anti-proliferation to breast cancer was therefore analyzed. GnRH-R expression in 298 primary breast cancer samples was measured by quantitative immunofluorescence. Levels of functional GnRH-R in breast-derived cell lines were assessed using 125 I-ligand binding and stimulation of 3 H-inositol phosphate production. Elevated levels of GnRH-R were stably expressed in cells by transfection. Effects of receptor activation on in vitro cell growth were investigated in comparison with IGF-I and EGF receptor inhibition, and correlated with intracellular signaling using western blotting. GnRH-R immunoscoring was highest in hormone receptor (triple) negative and grade 3 breast tumors. However prior to transfection, functional endogenous GnRH-R were undetectable in four commonly studied breast cancer cell lines (MCF-7, ZR-75-1, T47D and MDA-MB-231). After transfection with GnRH-R, high levels of cell surface GnRH-R were detected in SVCT and MDA-MB-231 clones while low-moderate levels of GnRH-R occurred in MCF-7 clones and ZR-75-1 clones. MCF-7 sub-clones with high levels of GnRH-R were isolated following hygromycin phosphotransferase transfection. High level cell surface GnRH-R enabled induction of high levels of 3 H-inositol phosphate and modest growth-inhibition in SVCT cells. In contrast, growth of MCF-7, ZR-75-1 or MDA-MB-231 clones was unaffected by GnRH-R activation. Cell growth was inhibited by IGF-I or EGF receptor inhibitors. IGF-I receptor inhibitor lowered levels of p-ERK1/2 in MCF-7 clones. Washout of IGF-I receptor inhibitor resulted in transient hyper-elevation of p-ERK1/2, but co-addition of GnRH-R agonist did not alter the dynamics of ERK1/2 re-phosphorylation. Breast cancers exhibit a range of GnRH-R immunostaining, with higher levels of

  2. Harnessing the potential of Knowledge Based Engineering in manufacturing design

    NARCIS (Netherlands)

    Van den Berg, T.

    2013-01-01

    An increase in productivity is required for the European aviation industry to remain competitive in a world with rising competition, shorter development timescales, more complex products and decreasing numbers of technically skilled personnel. Fokker Elmo, a Dutch aircraft electrical wiring harness

  3. Harnessing big data to meet the Sustainable Development Goals ...

    International Development Research Centre (IDRC) Digital Library (Canada)

    Harnessing big data to meet the Sustainable Development Goals – Building capacity in the Global South. Large volumes of complex and variable data, often called big data, promise to improve government service delivery, complement official statistics, and facilitate development in sectors such as health, urban ...

  4. Taso Mähar : avatud vannitoaga vanalinna korter / Gitte Hint

    Index Scriptorium Estoniae

    Hint, Gitte

    2003-01-01

    Modernne korter (220 m2). Sisearhitekt Taso Mähar, tema kommentaarid. Kogu korteri ulatuses on sisekujundusmaterjalidena kasutatud väärispuitu, metalli, marmorit ja klaasi. T. Mähari projekteeritud on trepp ja piirded, kamin, baarilett ja õhukesed alumiiniumraamidel mattklaasidega uksed. Ill.: I ja II korruse plaan, 8 värv. vaadet

  5. Harnessing the potentials of youth for rural household food security ...

    African Journals Online (AJOL)

    The food security question calls for participatory approach by the major stakeholders in the rural household in agriculture Youth are veritable assets in the household because of their dynamism, adventure, ambition and innovativeness. These potentials could be harnessed for rural household food security. This paper ...

  6. A Key Review of Non-Industrial Greywater Heat Harnessing

    Directory of Open Access Journals (Sweden)

    Abdur Rehman Mazhar

    2018-02-01

    Full Text Available The ever-growing concerns about making buildings more energy efficient and increasing the share of renewable energy used in them, has led to the development of ultra-low carbon buildings or passive houses. However, a huge potential still exists to lower the hot water energy demand, especially by harnessing heat from waste water exiting these buildings. Reusing this heat makes buildings more energy-efficient and this source is considered as a third-generation renewable energy technology, both factors conforming to energy policies throughout the world. Based on several theoretical and experimental studies, the potential to harness non-industrial waste water is quite high. As an estimate about 3.5 kWh of energy, per person per day could be harnessed and used directly, in many applications. A promising example of such an application, are low temperature fourth generation District Heating grids, with decentralized sources of heat. At the moment, heat exchangers and heat pumps are the only viable options to harness non-industrial waste heat. Both are used at different scales and levels of the waste-water treatment hierarchical pyramid. Apart from several unfavourable characteristics of these technologies, the associated exergetic efficiencies are low, in the range of 20–50%, even when cascaded combinations of both are used. To tackle these shortcomings, several promising trends and technologies are in the pipeline, to scavenge this small-scale source of heat to a large-scale benefit.

  7. Harnessing cell-material interaction to control cell fate: design ...

    Indian Academy of Sciences (India)

    Rajat K Das

    2017-10-26

    Oct 26, 2017 ... Harnessing cell-material interaction to control cell fate: design principle of advanced functional hydrogel materials. †. RAJAT K DAS. ∗. Materials Science Centre, Indian Institute of Technology, Kharagpur, West Bengal, India. E-mail: rajat@matsc.iitkgp.ernet.in. MS received 15 August 2017; revised 29 ...

  8. Harnessing the crowd to accelerate molecular medicine research.

    Science.gov (United States)

    Smith, Robert J; Merchant, Raina M

    2015-07-01

    Crowdsourcing presents a novel approach to solving complex problems within molecular medicine. By leveraging the expertise of fellow scientists across the globe, broadcasting to and engaging the public for idea generation, harnessing a scalable workforce for quick data management, and fundraising for research endeavors, crowdsourcing creates novel opportunities for accelerating scientific progress. Copyright © 2015 Elsevier Ltd. All rights reserved.

  9. HTLV-1 Tax mediated downregulation of miRNAs associated with chromatin remodeling factors in T cells with stably integrated viral promoter.

    Directory of Open Access Journals (Sweden)

    Saifur Rahman

    Full Text Available RNA interference (RNAi is a natural cellular mechanism to silence gene expression and is predominantly mediated by microRNAs (miRNAs that target messenger RNA. Viruses can manipulate the cellular processes necessary for their replication by targeting the host RNAi machinery. This study explores the effect of human T-cell leukemia virus type 1 (HTLV-1 transactivating protein Tax on the RNAi pathway in the context of a chromosomally integrated viral long terminal repeat (LTR using a CD4(+ T-cell line, Jurkat. Transcription factor profiling of the HTLV-1 LTR stably integrated T-cell clone transfected with Tax demonstrates increased activation of substrates and factors associated with chromatin remodeling complexes. Using a miRNA microarray and bioinformatics experimental approach, Tax was also shown to downregulate the expression of miRNAs associated with the translational regulation of factors required for chromatin remodeling. These observations were validated with selected miRNAs and an HTLV-1 infected T cells line, MT-2. miR-149 and miR-873 were found to be capable of directly targeting p300 and p/CAF, chromatin remodeling factors known to play critical role in HTLV-1 pathogenesis. Overall, these results are first in line establishing HTLV-1/Tax-miRNA-chromatin concept and open new avenues toward understanding retroviral latency and/or replication in a given cell type.

  10. HARNESS: Heterogeneous Adaptable Reconfigurable Networked Systems. Final Progress Report

    Energy Technology Data Exchange (ETDEWEB)

    Fagg, G. E.

    2004-01-20

    HARNESS was proposed as a system that combined the best of emerging technologies found in current distributed computing research and commercial products into a very flexible, dynamically adaptable framework that could be used by applications to allow them to evolve and better handle their execution environment. The HARNESS system was designed using the considerable experience from previous projects such as PVM, MPI, IceT and Cumulvs. As such, the system was designed to avoid any of the common problems found with using these current systems, such as no single point of failure, ability to survive machine, node and software failures. Additional features included improved intercomponent connectivity, with full support for dynamic down loading of addition components at run-time thus reducing the stress on application developers to build in all the libraries they need in advance.

  11. Uncovering Listeria monocytogenes hypervirulence by harnessing its biodiversity

    Science.gov (United States)

    Charlier, Caroline; Touchon, Marie; Chenal-Francisque, Viviane; Leclercq, Alexandre; Criscuolo, Alexis; Gaultier, Charlotte; Roussel, Sophie; Brisabois, Anne; Disson, Olivier; Rocha, Eduardo P. C.; Brisse, Sylvain; Lecuit, Marc

    2016-01-01

    Microbial pathogenesis studies are typically performed with reference strains, thereby overlooking microbial intra-species virulence heterogeneity. Here we integrated human epidemiological and clinical data with bacterial population genomics to harness the biodiversity of the model foodborne pathogen Listeria monocytogenes and decipher the basis of its neural and placental tropisms. Taking advantage of the clonal structure of this bacterial species, we identify clones epidemiologically associated with either food or human central nervous system (CNS) and maternal-neonatal (MN) listeriosis. The latter are also most prevalent in patients without immunosuppressive comorbidities. Strikingly, CNS and MN clones are hypervirulent in a humanized mouse model of listeriosis. By integrating epidemiological data and comparative genomics, we uncovered multiple novel putative virulence factors and demonstrated experimentally the contribution of the first gene cluster mediating Listeria monocytogenes neural and placental tropisms. This study illustrates the exceptional power of harnessing microbial biodiversity to identify clinically relevant microbial virulence attributes. PMID:26829754

  12. Uncovering Listeria monocytogenes hypervirulence by harnessing its biodiversity.

    Science.gov (United States)

    Maury, Mylène M; Tsai, Yu-Huan; Charlier, Caroline; Touchon, Marie; Chenal-Francisque, Viviane; Leclercq, Alexandre; Criscuolo, Alexis; Gaultier, Charlotte; Roussel, Sophie; Brisabois, Anne; Disson, Olivier; Rocha, Eduardo P C; Brisse, Sylvain; Lecuit, Marc

    2016-03-01

    Microbial pathogenesis studies are typically performed with reference strains, thereby overlooking within-species heterogeneity in microbial virulence. Here we integrated human epidemiological and clinical data with bacterial population genomics to harness the biodiversity of the model foodborne pathogen Listeria monocytogenes and decipher the basis of its neural and placental tropisms. Taking advantage of the clonal structure of this bacterial species, we identify clones epidemiologically associated either with food or with human central nervous system (CNS) or maternal-neonatal (MN) listeriosis. The latter clones are also most prevalent in patients without immunosuppressive comorbidities. Strikingly, CNS- and MN-associated clones are hypervirulent in a humanized mouse model of listeriosis. By integrating epidemiological data and comparative genomics, we have uncovered multiple new putative virulence factors and demonstrate experimentally the contribution of the first gene cluster mediating L. monocytogenes neural and placental tropisms. This study illustrates the exceptional power in harnessing microbial biodiversity to identify clinically relevant microbial virulence attributes.

  13. "Har andre plejebørn det som mig?"

    DEFF Research Database (Denmark)

    Warming, H.

    Bogen handler om livet som plejebarn, som det opleves af børnene selv. Den formidler resultaterne fra forskningsprojektet: "Projekt Børnetinget - et talerør for plejebørn", hvor 15 børn i familiepleje er blevet interviewet og hvor 107 nuværende og tidligere plejebørn har skrevet på hjemmesiden ww...

  14. Hello World: Harnessing social media for the Rosetta mission

    Science.gov (United States)

    Baldwin, E.; Mignone, C.; O'Flaherty, K. S.; Homfeld, A.-M.; Bauer, M.; McCaughrean, M. J.

    2015-10-01

    The European Space Agency's (ESA) comet-chasing Rosetta mission was launched in 2004, before social media became a popular tool for mainstream communication. By harnessing a range of platforms for communicating the key messages of this unprecedented space adventure as the spacecraft reached its destination ten years later, a wide range of new audiences were reached and could follow this once-in-a-lifetime mission.

  15. Harnessing the Power of Digital Data for Science and Society

    Science.gov (United States)

    2009-01-01

    Paul Ryan Department of Energy (DOE) George Seweryniak Walter Warnick Department of Homeland Security (DHS) Joseph Kielman Department of State Bie Yie...caltech.edu/~ george /sdt/sdt-final.pdf. 2 For purposes of this document, digital data are defined as any information that can be stored digitally and...Smithsonian Museums Yale Peabody Museum Exhibit C-1. Entities by Role HARNESSING THE POWER OF DIGITAL DATA FOR SCIENCE AND SOCIETY — C5 ENTITY TYPE ROLE

  16. Expression

    Directory of Open Access Journals (Sweden)

    Wang-Xia Wang

    2014-02-01

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

  17. HIV Provirus Stably Reproduces Parental Latent and Induced Transcription Phenotypes Regardless of the Chromosomal Integration Site.

    Science.gov (United States)

    Hashemi, Farhad B; Barreto, Kris; Bernhard, Wendy; Hashemi, Pargol; Lomness, Adam; Sadowski, Ivan

    2016-06-01

    Understanding the mechanisms of HIV proviral latency is essential for development of a means to eradicate infection and achieve a cure. We have previously described an in vitro latency model that reliably identifies HIV expression phenotypes of infected cells using a dual-fluorescence reporter virus. Our results have demonstrated that ∼50% of infected cells establish latency immediately upon integration of provirus, a phenomenon termed early latency, which appears to occur by mechanisms that are distinct from epigenetic silencing observed with HIV provirus that establishes productive infections. In this study, we have used a mini-dual HIV reporter virus (mdHIV) to compare the long-term stability of provirus produced as early latent or productive infections using Jurkat-Tat T cell clones. Cloned lines bearing mdHIV provirus integrated at different chromosomal locations display unique differences in responsiveness to signaling agonists and chromatin-modifying compounds, and they also produce characteristic expression patterns from the 5' long terminal repeat (LTR) dsRed and internal EIF1α-enhanced green fluorescent protein (EIF1α-eGFP) reporters. Furthermore, reporter expression profiles of single cell sorted subcultures faithfully reproduce expression profiles identical to that of their original parental population, following prolonged growth in culture, without shifting toward expression patterns resembling that of cell subclones at the time of sorting. Comparison of population dispersion coefficient (CV) and mean fluorescence intensity (MFI) of the subcloned lines showed that both untreated and phorbol myristate acetate (PMA)-ionomycin-stimulated cultures produce expression patterns identical to those of their parental lines. These results indicate that HIV provirus expression characteristics are strongly influenced by the epigenetic landscape at the site of chromosomal integration. There is currently considerable interest in development of therapies to

  18. Network harness: bundles of routes in public transport networks

    Science.gov (United States)

    Berche, B.; von Ferber, C.; Holovatch, T.

    2009-12-01

    Public transport routes sharing the same grid of streets and tracks are often found to proceed in parallel along shorter or longer sequences of stations. Similar phenomena are observed in other networks built with space consuming links such as cables, vessels, pipes, neurons, etc. In the case of public transport networks (PTNs) this behavior may be easily worked out on the basis of sequences of stations serviced by each route. To quantify this behavior we use the recently introduced notion of network harness. It is described by the harness distribution P(r, s): the number of sequences of s consecutive stations that are serviced by r parallel routes. For certain PTNs that we have analyzed we observe that the harness distribution may be described by power laws. These power laws indicate a certain level of organization and planning which may be driven by the need to minimize the costs of infrastructure and secondly by the fact that points of interest tend to be clustered in certain locations of a city. This effect may be seen as a result of the strong interdependence of the evolutions of both the city and its PTN. To further investigate the significance of the empirical results we have studied one- and two-dimensional models of randomly placed routes modeled by different types of walks. While in one dimension an analytic treatment was successful, the two dimensional case was studied by simulations showing that the empirical results for real PTNs deviate significantly from those expected for randomly placed routes.

  19. Construction of PVX virus-expression vector to express enterotoxin ...

    African Journals Online (AJOL)

    Potato X potyvirus (PVX)-based vector has been comprehensively applied in transient expression system. In order to produce the heterologous proteins more quickly and stably, the ClaI and NotI enzyme sites were introduced into the Enterotoxin fusion gene LTB-ST by polymerase chain reaction (PCR) and the LTB-ST ...

  20. Dielectric Spectroscopy and Optical Density Measurement for the Online Monitoring and Control of Recombinant Protein Production in Stably Transformed Drosophila melanogaster S2 Cells

    Directory of Open Access Journals (Sweden)

    Jan Zitzmann

    2018-03-01

    Full Text Available The production of recombinant proteins in bioreactors requires real-time process monitoring and control to increase process efficiency and to meet the requirements for a comprehensive audit trail. The combination of optical near-infrared turbidity sensors and dielectric spectroscopy provides diverse system information because different measurement principles are exploited. We used this combination of techniques to monitor and control the growth and protein production of stably transformed Drosophila melanogaster S2 cells expressing antimicrobial proteins. The in situ monitoring system was suitable in batch, fed-batch and perfusion modes, and was particularly useful for the online determination of cell concentration, specific growth rate (µ and cell viability. These data were used to pinpoint the optimal timing of the key transitional events (induction and harvest during batch and fed-batch cultivation, achieving a total protein yield of ~25 mg at the 1-L scale. During cultivation in perfusion mode, the OD880 signal was used to control the bleed line in order to maintain a constant cell concentration of 5 × 107 cells/mL, thus establishing a turbidostat/permittistat culture. With this setup, a five-fold increase in productivity was achieved and 130 mg of protein was recovered after 2 days of induced perfusion. Our results demonstrate that both sensors are suitable for advanced monitoring and integration into online control strategies.

  1. Harnessing Invariant NKT Cells to Improve Influenza Vaccines: A Pig Perspective

    Directory of Open Access Journals (Sweden)

    Guan Yang

    2017-12-01

    Full Text Available Invariant natural killer T (iNKT cells are an “innate-like” T cell lineage that recognize glycolipid rather than peptide antigens by their semi-invariant T cell receptors. Because iNKT cells can stimulate an extensive array of immune responses, there is considerable interest in targeting these cells to enhance human vaccines against a wide range of microbial pathogens. However, long overlooked is the potential to harness iNKT cell antigens as vaccine adjuvants for domestic animal species that express the iNKT cell–CD1d system. In this review, we discuss the prospect of targeting porcine iNKT cells as a strategy to enhance the efficiency of swine influenza vaccines. In addition, we compare the phenotype and tissue distribution of porcine iNKT cells. Finally, we discuss the challenges that must be overcome before iNKT cell agonists can be contemplated for veterinary use in livestock.

  2. Trunk Muscle Activation and Estimating Spinal Compressive Force in Rope and Harness Vertical Dance.

    Science.gov (United States)

    Wilson, Margaret; Dai, Boyi; Zhu, Qin; Humphrey, Neil

    2015-12-01

    Rope and harness vertical dance takes place off the floor with the dancer suspended from his or her center of mass in a harness attached to a rope from a point overhead. Vertical dance represents a novel environment for training and performing in which expected stresses on the dancer's body are different from those that take place during dance on the floor. Two male and eleven female dancers with training in vertical dance performed six typical vertical dance movements with electromyography (EMG) electrodes placed bilaterally on rectus abdominus, external oblique, erector spinae, and latissimus dorsi. EMG data were expressed as a percentage of maximum voluntary isometric contraction (MVIC). A simplified musculoskeletal model based on muscle activation for these four muscle groups was used to estimate the compressive force on the spine. The greatest muscle activation for erector spinae and latissimus dorsi and the greatest trunk compressive forces were seen in vertical axis positions where the dancer was moving the trunk into a hyper-extended position. The greatest muscle activation for rectus abdominus and external oblique and the second highest compressive force were seen in a supine position with the arms and legs extended away from the center of mass (COM). The least muscle activation occurred in positions where the limbs were hanging below the torso. These movements also showed relatively low muscle activation compression forces. Post-test survey results revealed that dancers felt comfortable in these positions; however, observation of some positions indicated insufficient muscular control. Computing the relative contribution of muscles, expressed as muscle activation and estimated spinal compression, provided a measure of how much the muscle groups were working to support the spine and the rest of the dancer's body in the different movements tested. Additionally, identifying typical muscle recruitment patterns in each movement will help identify key exercises

  3. How can developing countries harness biotechnology to improve health?

    Directory of Open Access Journals (Sweden)

    Persad Deepa L

    2007-12-01

    Full Text Available Abstract Background The benefits of genomics and biotechnology are concentrated primarily in the industrialized world, while their potential to combat neglected diseases in the developing world has been largely untapped. Without building developing world biotechnology capacity to address local health needs, this disparity will only intensify. To assess the potential of genomics to address health needs in the developing world, the McLaughlin-Rotman Centre for Global Health, along with local partners, organized five courses on Genomics and Public Health Policy in the developing world. The overall objective of the courses was to collectively explore how to best harness genomics to improve health in each region. This article presents and analyzes the recommendations from all five courses. Discussion In this paper we analyze recommendations from 232 developing world experts from 58 countries who sought to answer how best to harness biotechnology to improve health in their regions. We divide their recommendations into four categories: science; finance; ethics, society and culture; and politics. Summary The Courses' recommendations can be summarized across the four categories listed above: Science - Collaborate through national, regional, and international networks - Survey and build capacity based on proven models through education, training, and needs assessments Finance - Develop regulatory and intellectual property frameworks for commercialization of biotechnology - Enhance funding and affordability of biotechnology - Improve the academic-industry interface and the role of small and medium enterprise Ethics, Society, Culture - Develop public engagement strategies to inform and educate the public about developments in genomics and biotechnology - Develop capacity to address ethical, social and cultural issues - Improve accessibility and equity Politics - Strengthen understanding, leadership and support at the political level for biotechnology

  4. "Hello, World!" Harnessing Social Media for the Rosetta Mission

    Science.gov (United States)

    Baldwin, E.; Mignone, C.; Scuka, D.; Homfeld, A. M.; Ranero, K.; Rolfe, E.; Bennett, M.; Schepers, A.; O'Flaherty, K. S.; Bauer, M.; McCaughrean, M.

    2016-03-01

    The European Space Agency's comet-chasing Rosetta mission was launched in 2004, before social media became a popular tool for mainstream communication. As it reached its destination ten years later, new audiences were reached and inspired by this once-in-a-lifetime event by harnessing a range of outlets for communicating the key messages. These included traditional online platforms, such as news websites, blogs, and Livestream, as well as Twitter, Facebook, Instagram, Flickr, YouTube, Google+ and SoundCloud. In this article, we outline the role social media channels played in making Rosetta one of the European Space Agency's biggest communication and public engagement successes.

  5. Harnessing natural product assembly lines: structure, promiscuity, and engineering

    Science.gov (United States)

    Ladner, Christopher C; Williams, Gavin J

    2015-01-01

    Many therapeutically relevant natural products are biosynthesized by the action of giant mega-enzyme assembly lines. By leveraging the specificity, promiscuity, and modularity of assembly lines, a variety of strategies have been developed that enable the biosynthesis of modified natural products. This review briefly summarizes recent structural advances related to natural product assembly lines, discusses chemical approaches to probing assembly line structures in the absence of traditional biophysical data, and surveys efforts that harness the inherent or engineered promiscuity of assembly lines for the synthesis of non-natural polyketides and nonribosomal peptide analogues. PMID:26527577

  6. Harnessing natural product assembly lines: structure, promiscuity, and engineering.

    Science.gov (United States)

    Ladner, Christopher C; Williams, Gavin J

    2016-03-01

    Many therapeutically relevant natural products are biosynthesized by the action of giant mega-enzyme assembly lines. By leveraging the specificity, promiscuity, and modularity of assembly lines, a variety of strategies has been developed that enables the biosynthesis of modified natural products. This review briefly summarizes recent structural advances related to natural product assembly lines, discusses chemical approaches to probing assembly line structures in the absence of traditional biophysical data, and surveys efforts that harness the inherent or engineered promiscuity of assembly lines for the synthesis of non-natural polyketides and non-ribosomal peptide analogues.

  7. When sparks fly harnessing the power of group creativity

    CERN Document Server

    Leonard, Dorothy

    2005-01-01

    Creativity and innovation are evergreen topics. Particularly in today's tough marketplace, companies in every industry are looking for ways to harness creativity and innovation to spur growth and enhance competitiveness. This book differs from many books on creativity in that its focus is on how to manage a group, rather than individual creativity. It refutes much conventional wisdom about where creativity comes from, particularly that creativity does not stem from the random genius of individuals. The authors reveal specific group configurations and strategies that can unleash the collective creativity of any team or group.

  8. 21-Methylpyrenyl-cholesterol stably and specifically associates with lipoprotein peripheral hemi-membrane: A new labelling tool

    International Nuclear Information System (INIS)

    Gaibelet, Gérald; Tercé, François; Bertrand-Michel, Justine; Allart, Sophie; Azalbert, Vincent; Lecompte, Marie-France; Collet, Xavier; Orlowski, Stéphane

    2013-01-01

    Highlights: •21-Methylpyrenyl-cholesterol specifically and stably associates to lipoproteins. •It is not esterified by LCAT, and thus reliably labels their peripheral hemi-membrane. •HDL vs. LDL are well distinguishable by various fluorescent labelling characteristics. •LDL peripheral hemi-membrane harbors cholesterol-rich ordered lipid (micro)domains. •Cultured cells can be stained by such labelled lipoproteins-mediated delivery. -- Abstract: Lipoproteins are important biological components. However, they have few convenient fluorescent labelling probes currently reported, and their physiological reliability can be questioned. We compared the association of two fluorescent cholesterol derivatives, 22-nitrobenzoxadiazole-cholesterol (NBD-Chol) and 21-methylpyrenyl-cholesterol (Pyr-met-Chol), to serum lipoproteins and to purified HDL and LDL. Both lipoproteins could be stably labelled by Pyr-met-Chol, but virtually not by NBD-Chol. At variance with NBD-Chol, LCAT did not esterify Pyr-met-Chol. The labelling characteristics of lipoproteins by Pyr-met-Chol were well distinguishable between HDL and LDL, regarding dializability, associated probe amount and labelling kinetics. We took benefit of the pyrene labelling to approach the structural organization of LDL peripheral hemi-membrane, since Pyr-met-Chol-labelled LDL, but not HDL, presented a fluorescence emission of pyrene excimers, indicating that the probe was present in an ordered lipid micro-environment. Since the peripheral membrane of LDL contains more sphingomyelin (SM) than HDL, this excimer formation was consistent with the existence of cholesterol- and SM-enriched lipid microdomains in LDL, as already suggested in model membranes of similar composition and reminiscent to the well-described “lipid rafts” in bilayer membranes. Finally, we showed that Pyr-met-Chol could stain cultured PC-3 cells via lipoprotein-mediated delivery, with a staining pattern well different to that observed with NBD

  9. 21-Methylpyrenyl-cholesterol stably and specifically associates with lipoprotein peripheral hemi-membrane: A new labelling tool

    Energy Technology Data Exchange (ETDEWEB)

    Gaibelet, Gérald [INSERM U563, CHU Purpan, Toulouse (France); CEA, SB2SM and UMR8221 CNRS, IBiTec-Saclay, Gif-sur-Yvette (France); Tercé, François [Université Toulouse III, UMR 1048, Toulouse (France); INSERM U1048, Toulouse (France); Bertrand-Michel, Justine [Université Toulouse III, UMR 1048, Toulouse (France); INSERM U1048, Lipidomic Platform Metatoul, Toulouse (France); Allart, Sophie [Plateau Technique d’Imagerie Cellulaire, INSERM U1043, Toulouse (France); Azalbert, Vincent [Université Toulouse III, UMR 1048, Toulouse (France); INSERM U1048, Toulouse (France); Lecompte, Marie-France [INSERM U563, Faculté de Médecine de Rangueil, Toulouse (France); Collet, Xavier [Université Toulouse III, UMR 1048, Toulouse (France); INSERM U1048, Toulouse (France); Orlowski, Stéphane, E-mail: stephane.orlowski@cea.fr [INSERM U563, CHU Purpan, Toulouse (France); CEA, SB2SM and UMR8221 CNRS, IBiTec-Saclay, Gif-sur-Yvette (France)

    2013-11-01

    Highlights: •21-Methylpyrenyl-cholesterol specifically and stably associates to lipoproteins. •It is not esterified by LCAT, and thus reliably labels their peripheral hemi-membrane. •HDL vs. LDL are well distinguishable by various fluorescent labelling characteristics. •LDL peripheral hemi-membrane harbors cholesterol-rich ordered lipid (micro)domains. •Cultured cells can be stained by such labelled lipoproteins-mediated delivery. -- Abstract: Lipoproteins are important biological components. However, they have few convenient fluorescent labelling probes currently reported, and their physiological reliability can be questioned. We compared the association of two fluorescent cholesterol derivatives, 22-nitrobenzoxadiazole-cholesterol (NBD-Chol) and 21-methylpyrenyl-cholesterol (Pyr-met-Chol), to serum lipoproteins and to purified HDL and LDL. Both lipoproteins could be stably labelled by Pyr-met-Chol, but virtually not by NBD-Chol. At variance with NBD-Chol, LCAT did not esterify Pyr-met-Chol. The labelling characteristics of lipoproteins by Pyr-met-Chol were well distinguishable between HDL and LDL, regarding dializability, associated probe amount and labelling kinetics. We took benefit of the pyrene labelling to approach the structural organization of LDL peripheral hemi-membrane, since Pyr-met-Chol-labelled LDL, but not HDL, presented a fluorescence emission of pyrene excimers, indicating that the probe was present in an ordered lipid micro-environment. Since the peripheral membrane of LDL contains more sphingomyelin (SM) than HDL, this excimer formation was consistent with the existence of cholesterol- and SM-enriched lipid microdomains in LDL, as already suggested in model membranes of similar composition and reminiscent to the well-described “lipid rafts” in bilayer membranes. Finally, we showed that Pyr-met-Chol could stain cultured PC-3 cells via lipoprotein-mediated delivery, with a staining pattern well different to that observed with NBD

  10. Stably Integrated luxCDABE for Assessment of Salmonella Invasion Kinetics

    Directory of Open Access Journals (Sweden)

    Kelly N. Flentie

    2008-09-01

    Full Text Available Salmonella Typhimurium is a common cause of gastroenteritis in humans and also localizes to neoplastic tumors in animals. Invasion of specific eukaryotic cells is a key mechanism of Salmonella interactions with host tissues. Early stages of gastrointestinal cell invasion are mediated by a Salmonella type III secretion system, powered by the adenosine triphosphatase invC. The aim of this work was to characterize the invC dependence of invasion kinetics into disparate eukaryotic cells traditionally used as models of gut epithelium or neoplasms. Thus, a nondestructive real-time assay was developed to report eukaryotic cell invasion kinetics using lux+ Salmonella that contain chromosomally integrated luxCDABE genes. Bioluminescence-based invasion assays using lux+ Salmonella exhibited inoculum dose-response correlation, distinguished invasion-competent from invasion-incompetent Salmonella, and discriminated relative Salmonella invasiveness in accordance with environmental conditions that induce invasion gene expression. In standard gentamicin protection assays, bioluminescence from lux+ Salmonella correlated with recovery of colony-forming units of internalized bacteria and could be visualized by bioluminescence microscopy. Furthermore, this assay distinguished invasion-competent from invasion-incompetent bacteria independent of gentamicin treatment in real time. Bioluminescence reported Salmonella invasion of disparate eukaryotic cell lines, including neoplastic melanoma, colon adenocarcinoma, and glioma cell lines used in animal models of malignancy. In each case, Salmonella invasion of eukaryotic cells was invC dependent.

  11. Synthesis of Fluorescent Carbon Dots by Gastrointestinal Fluid Treatment of Mongolia Har Gabur

    Directory of Open Access Journals (Sweden)

    Tegexibaiyin Wang

    2017-01-01

    Full Text Available Har Gabur is the carbide obtained from pig manure by burning. The fluorescent carbon dots (CDs of Har Gabur were successfully synthesized through simulating the digestion process of human gastrointestinal tract. Transmission Electron Microscope (TEM analysis showed that the average size of the prepared Har Gabur CDs was 4 nm, with good solubility in water and strong fluorescence under UV irradiation. The X-ray and Raman results showed that the Har Gabur CDs were mainly composed of oxygen “O” and carbon “C” elements, in the forms of “C=O” and “C-O.” The bond energy results showed that the nitrogen “N” atom presented as “C-N” form, which indicated that Har Gabur CDs also contain “N.” In photobleaching assay, Har Gabur CDs showed excellent light stability compared with ordinary organic dye, fluorescein, and Rhodamine B. The fluorescence intensity of Har Gabur CDs was fairly stable within a wide pH range of 3–10. When L-lysine and L-cysteine were applied for the passivation stage, the relative quantum yields were improved by 1.53 and 3.68 times, respectively. Finally, the fluorescence properties of Har Gabur CDs were tested in cells and zebrafish, illustrating that Har Gabur CD has potential in the application of biological labeling and imaging.

  12. A test harness for accelerating physics parameterization advancements into operations

    Science.gov (United States)

    Firl, G. J.; Bernardet, L.; Harrold, M.; Henderson, J.; Wolff, J.; Zhang, M.

    2017-12-01

    The process of transitioning advances in parameterization of sub-grid scale processes from initial idea to implementation is often much quicker than the transition from implementation to use in an operational setting. After all, considerable work must be undertaken by operational centers to fully test, evaluate, and implement new physics. The process is complicated by the scarcity of like-to-like comparisons, availability of HPC resources, and the ``tuning problem" whereby advances in physics schemes are difficult to properly evaluate without first undertaking the expensive and time-consuming process of tuning to other schemes within a suite. To address this process shortcoming, the Global Model TestBed (GMTB), supported by the NWS NGGPS project and undertaken by the Developmental Testbed Center, has developed a physics test harness. It implements the concept of hierarchical testing, where the same code can be tested in model configurations of varying complexity from single column models (SCM) to fully coupled, cycled global simulations. Developers and users may choose at which level of complexity to engage. Several components of the physics test harness have been implemented, including a SCM and an end-to-end workflow that expands upon the one used at NOAA/EMC to run the GFS operationally, although the testbed components will necessarily morph to coincide with changes to the operational configuration (FV3-GFS). A standard, relatively user-friendly interface known as the Interoperable Physics Driver (IPD) is available for physics developers to connect their codes. This prerequisite exercise allows access to the testbed tools and removes a technical hurdle for potential inclusion into the Common Community Physics Package (CCPP). The testbed offers users the opportunity to conduct like-to-like comparisons between the operational physics suite and new development as well as among multiple developments. GMTB staff have demonstrated use of the testbed through a

  13. A New Harness For Use with Exercise Countermeasures-Validation of Improved Comfort and Loading with the Center for Space Medicine (CSM) Harness

    Data.gov (United States)

    National Aeronautics and Space Administration — The on-orbit treadmill harness Station Development Test Objective (SDTO) evaluation ran from Increment 21 through 25 aboard the International Space Station...

  14. Spil har potentiale som motivation for læring

    DEFF Research Database (Denmark)

    Hedegaard, Lars Norup

    En designbaseret empirisk undersøgelse af almindeligt anvendte gamification elementer. Gamification har til formål at skabe fornøjelse, samt en følelse af leg, i ikke-spil miljøer. Grundet en øget brug af gamification på arbejdspladser og i læreprocesser ses området vigtigt at undersøge yderligere....... Projektet søger at belyse, hvordan gamification påvirker motivationen for læring, herunder hvordan almindeligt anvendte spil-elementer og -genrer kan indgå i planlægningen af læringsaktiviteter. Projektet tager omdrejningspunkt i de studerendes oplevede erfaringer, med den antagelse, at intervention i...

  15. Når sex er en ret, hvem har så pligt?

    DEFF Research Database (Denmark)

    Munk-Madsen, Eva

    2007-01-01

    Socialministeriet har fastslået i en vejledning, at samfundet har omsorgspligt for funktionshæmmedes borgeres  seksuelle udfoldelsesmulighder. Tre grupper er udpeget som dem der skal varetage denne omsorgspligt på samfundets vegne: ansatte hjælpepersoner, seksualvejledere og prostituerede. I modt...

  16. Vi har brug for dystopierne - men de kan ikke stå alene

    DEFF Research Database (Denmark)

    Baron, Christian

    2017-01-01

    Filmatiseringen af Margaret Atwoods »The Handmaid’s Tale« har aktualiseret dystopiens potentiale som samfundskritisk betydningsbærer. I sagens natur er dystopiens evne til at formulere alternativer til den udvikling, den advarer imod, imidlertid begrænsede. Vi har brug for litteratur, der tager...

  17. THE EVALUATION OF PEPTIDE/HISTIDINE TRANSPORTER 1 (PHT1) FUNCTION: UPTAKE KINETICS UTILIZING A COS-7 STABLY TRANSFECTED CELL LINE.

    Science.gov (United States)

    Lindley, David J; Carl, Stephen M; Mowery, Stephanie A; Knipp, Gregory T

    2011-10-01

    There have been relatively few studies focused on the proton-dependent oligopeptide transporter (POT) superfamily member, Peptide/Histidine Transporter 1 (PHT1), with respect to its contribution to the ADME of peptides and peptide-based drugs. These studies were conducted to determine hPHT1-mediated, H + -dependent uptake kinetics of histidine, carnosine, Gly-Sar and valacyclovir in stably transfected hPHT1-COS-7 cells comparative to kinetics determined in an empty vector (Mock) stably transfected cell line. The results suggest that Gly-Sar appears to be a substrate for PHT1 based on efflux from the stably transfected hPHT1 COS-7 cells. Histidine and Gly-Sar concentration- and time-dependent studies suggest mixed-uptake kinetics. These studies suggest that stably transfected hPHT1-COS-7 cells exhibit different uptake kinetics than those observed in our previous studies and illustrate the requirement for experiments to delineate the physiological role of hPHT1.

  18. Invariant Natural Killer T Cells in Immune Regulation of Blood Cancers: Harnessing Their Potential in Immunotherapies

    Directory of Open Access Journals (Sweden)

    Pui Yeng Lam

    2017-10-01

    Full Text Available Invariant natural killer T (iNKT cells are a unique innate T lymphocyte population that possess cytolytic properties and profound immunoregulatory activities. iNKT cells play an important role in the immune surveillance of blood cancers. They predominantly recognize glycolipid antigens presented on CD1d, but their activation and cytolytic activities are not confined to CD1d expressing cells. iNKT cell stimulation and subsequent production of immunomodulatory cytokines serve to enhance the overall antitumor immune response. Crucially, the activation of iNKT cells in cancer often precedes the activation and priming of other immune effector cells, such as NK cells and T cells, thereby influencing the generation and outcome of the antitumor immune response. Blood cancers can evade or dampen iNKT cell responses by downregulating expression of recognition receptors or by actively suppressing or diverting iNKT cell functions. This review will discuss literature on iNKT cell activity and associated dysregulation in blood cancers as well as highlight some of the strategies designed to harness and enhance iNKT cell functions against blood cancers.

  19. Bioactive polyphenols from muscadine grape and blackcurrant stably concentrated onto protein-rich matrices for topical applications.

    Science.gov (United States)

    Plundrich, N; Grace, M H; Raskin, I; Ann Lila, M

    2013-08-01

    Natural botanical agents that are antimicrobial, or that modulate skin hyperpigmentation via tyrosinase inhibition, are increasingly sought in the cosmetic industry. In this study, an efficient tactic is demonstrated for concentrating and stabilizing skin-beneficial bioactive compounds from muscadine grape and blackcurrant juice or muscadine pomace, into hemp flour (HF), hemp protein isolate (HPI) and soy protein isolate (SPI) matrices suitable for cosmetic applications. Anthocyanins were most efficiently captured from blackcurrant juice into HF (8.39 mg g(-1) ). HPI most effectively captured total phenolics from muscadine pomace (72.32 and 77.32 mg g(-1) from Noble and Carlos, respectively), while the three matrices incorporated highest levels of ellagic acid, gallic acid, and PAC B1 from Noble muscadine grape juice. The enriched matrices demonstrated effective in vitro inhibition of tyrosinase (up to 57.29% for blackcurrant juice-HPI matrix), and in general, juice sources provided greater inhibition on L-dopamine oxidation by tyrosinase than pomace sources. The polyphenol-enriched matrices effectively inhibited microbial proliferation in a screening assay against Staphylococcus aureus bacteria, whereas untreated HF, HPI or SPI did not inhibit bacterial growth. The technology of combining and stably concentrating phytoactive polyphenols with proteins has potential use for cosmetic topical applications. © 2013 John Wiley & Sons Ltd.

  20. Harnessing diversity towards the reconstructing of large scale gene regulatory networks.

    Directory of Open Access Journals (Sweden)

    Takeshi Hase

    Full Text Available Elucidating gene regulatory network (GRN from large scale experimental data remains a central challenge in systems biology. Recently, numerous techniques, particularly consensus driven approaches combining different algorithms, have become a potentially promising strategy to infer accurate GRNs. Here, we develop a novel consensus inference algorithm, TopkNet that can integrate multiple algorithms to infer GRNs. Comprehensive performance benchmarking on a cloud computing framework demonstrated that (i a simple strategy to combine many algorithms does not always lead to performance improvement compared to the cost of consensus and (ii TopkNet integrating only high-performance algorithms provide significant performance improvement compared to the best individual algorithms and community prediction. These results suggest that a priori determination of high-performance algorithms is a key to reconstruct an unknown regulatory network. Similarity among gene-expression datasets can be useful to determine potential optimal algorithms for reconstruction of unknown regulatory networks, i.e., if expression-data associated with known regulatory network is similar to that with unknown regulatory network, optimal algorithms determined for the known regulatory network can be repurposed to infer the unknown regulatory network. Based on this observation, we developed a quantitative measure of similarity among gene-expression datasets and demonstrated that, if similarity between the two expression datasets is high, TopkNet integrating algorithms that are optimal for known dataset perform well on the unknown dataset. The consensus framework, TopkNet, together with the similarity measure proposed in this study provides a powerful strategy towards harnessing the wisdom of the crowds in reconstruction of unknown regulatory networks.

  1. Effects of recombinant plasmid pEgr-p53 transfected stably in combination with X-irradiation on cell cycle progression and proliferation in human SKOV-3 tumor cells in vitro

    International Nuclear Information System (INIS)

    Dong Lihua; Liu Feng; Li Yanbo; Fu Shibo; Gong Shouliang

    2008-01-01

    Objective: To investigate the effect of recombinant plasmid pEgr-hp53 transfected stably in combination with X-ray irradiation on the cell cycle progression and the proliferation in human SKOV-3 tumor cells. Methods: pEgr-hp53 and pcDNA3.1 packaged with liposome were stably transfected into SKOV-3 cells in vitro. SKOV-3-hp53 and SKOV-3-vect were irradiated with 0, 0.5, 2.0 and 5.0 Gy X-rays, respectively, i.e. 8 experimental groups. The SKOV-3 cell proliferation and the cell cycle progression were measured with flow cytometry and cell growth curve, respectively. Results: Compared with 0 Gy group, the cell counts in SKOV-3- hp53 plus different doses of irradiation groups 2 d after irradiation decreased significantly (P 0 /G 1 cells increased significantly (P 2 /M cells decreased in varying degrees. The cell counts in SKOV-3-hp53 plus irradiation group were significantly lower than those in corresponding SKOV-3-vect plus irradiation group, the cell counts 4-8 d after irradiation with 0.5 Gy, 2 d after 2.0 Gy irradiation and 6 d after 5.0 Gy irradiation decreased significantly (P 0 /G 1 cells increased significantly (P 2 /M cells decreased significantly (P 1 arrest in SKOV-3 cells and inhibits the cell proliferation. Ionizing radiation can activate early growth response-1 (Egr-1) gene promoter and increase the expression of p53 gene, and enhance the inhibition of tumor cell growth. (authors)

  2. Health as foreign policy: harnessing globalization for health.

    Science.gov (United States)

    Fidler, David P

    2006-12-01

    This paper explores the importance for health promotion of the rise of public health as a foreign policy issue. Although health promotion encompassed foreign policy as part of 'healthy public policy', mainstream foreign policy neglected public health and health promotion's role in it. Globalization forces health promotion, however, to address directly the relationship between public health and foreign policy. The need for 'health as foreign policy' is apparent from the prominence public health now has in all the basic governance functions served by foreign policy. The Secretary-General's United Nations (UN) reform proposals demonstrate the importance of foreign policy to health promotion as a core component of public health because the proposals embed public health in each element of the Secretary-General's vision for the UN in the 21st century. The emergence of health as foreign policy presents opportunities and risks for health promotion that can be managed by emphasizing that public health constitutes an integrated public good that benefits all governance tasks served by foreign policy. Any effort to harness globalization for public health will have to make health as foreign policy a centerpiece of its ambitions, and this task is now health promotion's burden and opportunity.

  3. The globalization of health research: harnessing the scientific diaspora.

    Science.gov (United States)

    Anand, Nalini P; Hofman, Karen J; Glass, Roger I

    2009-04-01

    The scientific diaspora is a unique resource for U.S. universities. By drawing on the expertise, experience, and catalytic potential of diaspora scientists, universities can capitalize more fully on their diverse intellectual resources to make lasting contributions to global health. This article examines the unique contributions of the diaspora in international research collaborations, advantages of harnessing the diaspora and benefits to U.S. universities of fostering these collaborations, challenges faced by scientists who want to work with their home countries, examples of scientists engaging with their home countries, and specific strategies U.S. universities and donors can implement to catalyze these collaborations. The contributions of the diaspora to the United States are immense: International students enrolled in academic year 2007-2008 contributed an estimated $15 billion to the U.S. economy. As scientific research becomes increasingly global, the percentage of scientific publications with authors from foreign countries has grown from 8% in 1988 to 20% in 2005. Diaspora scientists can help build trusting relationships with scientists abroad, and international collaborations may improve the health of underserved populations at home. Although opportunities for diaspora networks are increasing, most home countries often lack enabling policies, infrastructure, and resources to effectively utilize their diaspora communities abroad. This article examines how some governments have successfully mobilized their scientific diaspora to become increasingly engaged in their national research agendas. Recommendations include specific strategies, including those that encourage U.S. universities to promote mini-sabbaticals and provide seed funding and flexible time frames.

  4. Harnessing the Therapeutic Potential of Th17 Cells

    Directory of Open Access Journals (Sweden)

    Jonas Bystrom

    2015-01-01

    Full Text Available Th17 cells provide protective immunity to infections by fungi and extracellular bacteria as well as cancer but are also involved in chronic inflammation. The cells were first identified by their ability to produce interleukin 17A (IL-17A and, subsequently, associated with chronic inflammation and autoimmunity. Th17 cells have some gene profile similarity with stem cells and can remain dormant in mucosal tissues for long periods. Indeed, recent studies suggest that functionally distinct subsets of pro- and anti-inflammatory Th17 cells can interchange phenotype and functions. For development, Th17 cells require activation of the transcription factors STAT3 and RORγt while RUNX1, c-Maf, and Aiolos are involved in changes of phenotype/functions. Attempts to harness Th17 cells against pathogens and cancer using vaccination strategies are being explored. The cells gain protective abilities when induced to produce interferon γ (IFNγ. In addition, treatment with antibodies to IL-17 is effective in treating patients with psoriasis, psoriatic arthritis, and refectory rheumatoid arthritis. Moreover, since RORγt is a nuclear receptor, it is likely to be a potential future drug target for modulating Th17 functions. This review explores pathways through which Th17 subsets are induced, the molecular basis of their plasticity, and potential therapeutic strategies for their modulation in diseases.

  5. Weakly supervised visual dictionary learning by harnessing image attributes.

    Science.gov (United States)

    Gao, Yue; Ji, Rongrong; Liu, Wei; Dai, Qionghai; Hua, Gang

    2014-12-01

    Bag-of-features (BoFs) representation has been extensively applied to deal with various computer vision applications. To extract discriminative and descriptive BoF, one important step is to learn a good dictionary to minimize the quantization loss between local features and codewords. While most existing visual dictionary learning approaches are engaged with unsupervised feature quantization, the latest trend has turned to supervised learning by harnessing the semantic labels of images or regions. However, such labels are typically too expensive to acquire, which restricts the scalability of supervised dictionary learning approaches. In this paper, we propose to leverage image attributes to weakly supervise the dictionary learning procedure without requiring any actual labels. As a key contribution, our approach establishes a generative hidden Markov random field (HMRF), which models the quantized codewords as the observed states and the image attributes as the hidden states, respectively. Dictionary learning is then performed by supervised grouping the observed states, where the supervised information is stemmed from the hidden states of the HMRF. In such a way, the proposed dictionary learning approach incorporates the image attributes to learn a semantic-preserving BoF representation without any genuine supervision. Experiments in large-scale image retrieval and classification tasks corroborate that our approach significantly outperforms the state-of-the-art unsupervised dictionary learning approaches.

  6. Flow Control in Wells Turbines for Harnessing Maximum Wave Power.

    Science.gov (United States)

    Lekube, Jon; Garrido, Aitor J; Garrido, Izaskun; Otaola, Erlantz; Maseda, Javier

    2018-02-10

    Oceans, and particularly waves, offer a huge potential for energy harnessing all over the world. Nevertheless, the performance of current energy converters does not yet allow us to use the wave energy efficiently. However, new control techniques can improve the efficiency of energy converters. In this sense, the plant sensors play a key role within the control scheme, as necessary tools for parameter measuring and monitoring that are then used as control input variables to the feedback loop. Therefore, the aim of this work is to manage the rotational speed control loop in order to optimize the output power. With the help of outward looking sensors, a Maximum Power Point Tracking (MPPT) technique is employed to maximize the system efficiency. Then, the control decisions are based on the pressure drop measured by pressure sensors located along the turbine. A complete wave-to-wire model is developed so as to validate the performance of the proposed control method. For this purpose, a novel sensor-based flow controller is implemented based on the different measured signals. Thus, the performance of the proposed controller has been analyzed and compared with a case of uncontrolled plant. The simulations demonstrate that the flow control-based MPPT strategy is able to increase the output power, and they confirm both the viability and goodness.

  7. Cre recombinase expression can result in phenotypic aberrations in plants

    NARCIS (Netherlands)

    Coppoolse, E.; Vroomen, de M.J.; Roelofs, D.; Smit, J.; Gennip, van F.; Hersmus, B.J.M.; Nijkamp, H.J.J.; Haaren, van M.J.

    2003-01-01

    The cre recombinase gene was stably introduced and expressed in tomato, petunia and Nicotiana tabacum. Some plants expressing the cre gene driven by a CaMV 35S promoter displayed growth retardation and a distinct pattern of chlorosis in their leaves. Although no direct relation can be proven between

  8. Cre recombinase expression can result in phenotypic aberrations in plants

    NARCIS (Netherlands)

    Coppoolse, Eric R; de Vroomen, Marianne J; Roelofs, Dick; Smit, Jaap; van Gennip, Femke; Hersmus, Bart J M; Nijkamp, H John J; van Haaren, Mark J J

    The cre recombinase gene was stably introduced and expressed in tomato, petunia and Nicotiana tabacum. Some plants expressing the cre gene driven by a CaMV 35S promoter displayed growth retardation and a distinct pattern of chlorosis in their leaves. Although no direct relation can be proven between

  9. Construction of stably maintained non-mobilizable derivatives of RSF1010 lacking all known elements essential for mobilization

    Directory of Open Access Journals (Sweden)

    Tokmakova Irina L

    2007-11-01

    Full Text Available Abstract Background RSF1010 is a well-studied broad-host-range plasmid able to be mobilized to different bacteria and plants. RSF1010-derived plasmid vectors are widely used in both basic research and industrial applications. In the latter case, exploiting of mobilizable plasmids or even the plasmids possessing negligible mobilization frequency, but containing DNA fragments that could promote conjugal transfer, is undesirable because of biosafety considerations. Previously, several mutations significantly decreasing efficiency of RSF1010 mobilization have been selected. Nevertheless, construction of the RSF1010 derivative lacking all known loci involved in the conjugal transfer has not been reported yet. Results Novel non-mobilizable derivatives of RSF1010 lacking all known DNA sequences involved in the mobilization process have been obtained due to the exploiting of λRed-driven recombination between the plasmid and a constructed in vitro linear DNA fragment. To provide auto-regulated transcription of the essential replication gene, repB, the plasmid loci oriT, mobC and mobA were substituted by the DNA fragment containing PlacUV5→lacI. Mobilization of the obtained RSFmob plasmid was not detected in standard tests. The derivative of RSFmob with increased copy number has been obtained after lacI elimination. High stability of both constructed plasmids has been demonstrated in Escherichia coli and Pantoea ananatis. Design of RSFmob allows easy substitution of PlacUV5 by any desirable promoter for construction of novel derivatives with changed copy number or host range. Conclusion Novel non-mobilizable derivatives of RSF1010 lacking all known DNA sequences involved in the mobilization process and stably maintained at least in E. coli and P. ananatis have been constructed. The obtained plasmids became the progenitors of new cloning vectors answering all biosafety requirements of genetically modified organisms used in scale-up production.

  10. Harnessing mosquito-Wolbachia symbiosis for vector and disease control.

    Science.gov (United States)

    Bourtzis, Kostas; Dobson, Stephen L; Xi, Zhiyong; Rasgon, Jason L; Calvitti, Maurizio; Moreira, Luciano A; Bossin, Hervé C; Moretti, Riccardo; Baton, Luke Anthony; Hughes, Grant L; Mavingui, Patrick; Gilles, Jeremie R L

    2014-04-01

    Mosquito species, members of the genera Aedes, Anopheles and Culex, are the major vectors of human pathogens including protozoa (Plasmodium sp.), filariae and of a variety of viruses (causing dengue, chikungunya, yellow fever, West Nile). There is lack of efficient methods and tools to treat many of the diseases caused by these major human pathogens, since no efficient vaccines or drugs are available; even in malaria where insecticide use and drug therapies have reduced incidence, 219 million cases still occurred in 2010. Therefore efforts are currently focused on the control of vector populations. Insecticides alone are insufficient to control mosquito populations since reduced susceptibility and even resistance is being observed more and more frequently. There is also increased concern about the toxic effects of insecticides on non-target (even beneficial) insect populations, on humans and the environment. During recent years, the role of symbionts in the biology, ecology and evolution of insect species has been well-documented and has led to suggestions that they could potentially be used as tools to control pests and therefore diseases. Wolbachia is perhaps the most renowned insect symbiont, mainly due to its ability to manipulate insect reproduction and to interfere with major human pathogens thus providing new avenues for pest control. We herein present recent achievements in the field of mosquito-Wolbachia symbiosis with an emphasis on Aedes albopictus. We also discuss how Wolbachia symbiosis can be harnessed for vector control as well as the potential to combine the sterile insect technique and Wolbachia-based approaches for the enhancement of population suppression programs. Copyright © 2013 International Atomic Energy Agency 2013. Published by Elsevier B.V. All rights reserved.

  11. Identification and validation of suitable endogenous reference genes for gene expression studies in human peripheral blood.

    Science.gov (United States)

    Stamova, Boryana S; Apperson, Michelle; Walker, Wynn L; Tian, Yingfang; Xu, Huichun; Adamczy, Peter; Zhan, Xinhua; Liu, Da-Zhi; Ander, Bradley P; Liao, Isaac H; Gregg, Jeffrey P; Turner, Renee J; Jickling, Glen; Lit, Lisa; Sharp, Frank R

    2009-08-05

    Gene expression studies require appropriate normalization methods. One such method uses stably expressed reference genes. Since suitable reference genes appear to be unique for each tissue, we have identified an optimal set of the most stably expressed genes in human blood that can be used for normalization. Whole-genome Affymetrix Human 2.0 Plus arrays were examined from 526 samples of males and females ages 2 to 78, including control subjects and patients with Tourette syndrome, stroke, migraine, muscular dystrophy, and autism. The top 100 most stably expressed genes with a broad range of expression levels were identified. To validate the best candidate genes, we performed quantitative RT-PCR on a subset of 10 genes (TRAP1, DECR1, FPGS, FARP1, MAPRE2, PEX16, GINS2, CRY2, CSNK1G2 and A4GALT), 4 commonly employed reference genes (GAPDH, ACTB, B2M and HMBS) and PPIB, previously reported to be stably expressed in blood. Expression stability and ranking analysis were performed using GeNorm and NormFinder algorithms. Reference genes were ranked based on their expression stability and the minimum number of genes needed for nomalization as calculated using GeNorm showed that the fewest, most stably expressed genes needed for acurate normalization in RNA expression studies of human whole blood is a combination of TRAP1, FPGS, DECR1 and PPIB. We confirmed the ranking of the best candidate control genes by using an alternative algorithm (NormFinder). The reference genes identified in this study are stably expressed in whole blood of humans of both genders with multiple disease conditions and ages 2 to 78. Importantly, they also have different functions within cells and thus should be expressed independently of each other. These genes should be useful as normalization genes for microarray and RT-PCR whole blood studies of human physiology, metabolism and disease.

  12. Identification and validation of suitable endogenous reference genes for gene expression studies in human peripheral blood

    Directory of Open Access Journals (Sweden)

    Turner Renee J

    2009-08-01

    Full Text Available Abstract Background Gene expression studies require appropriate normalization methods. One such method uses stably expressed reference genes. Since suitable reference genes appear to be unique for each tissue, we have identified an optimal set of the most stably expressed genes in human blood that can be used for normalization. Methods Whole-genome Affymetrix Human 2.0 Plus arrays were examined from 526 samples of males and females ages 2 to 78, including control subjects and patients with Tourette syndrome, stroke, migraine, muscular dystrophy, and autism. The top 100 most stably expressed genes with a broad range of expression levels were identified. To validate the best candidate genes, we performed quantitative RT-PCR on a subset of 10 genes (TRAP1, DECR1, FPGS, FARP1, MAPRE2, PEX16, GINS2, CRY2, CSNK1G2 and A4GALT, 4 commonly employed reference genes (GAPDH, ACTB, B2M and HMBS and PPIB, previously reported to be stably expressed in blood. Expression stability and ranking analysis were performed using GeNorm and NormFinder algorithms. Results Reference genes were ranked based on their expression stability and the minimum number of genes needed for nomalization as calculated using GeNorm showed that the fewest, most stably expressed genes needed for acurate normalization in RNA expression studies of human whole blood is a combination of TRAP1, FPGS, DECR1 and PPIB. We confirmed the ranking of the best candidate control genes by using an alternative algorithm (NormFinder. Conclusion The reference genes identified in this study are stably expressed in whole blood of humans of both genders with multiple disease conditions and ages 2 to 78. Importantly, they also have different functions within cells and thus should be expressed independently of each other. These genes should be useful as normalization genes for microarray and RT-PCR whole blood studies of human physiology, metabolism and disease.

  13. Harnessing light: optical science and engineering for the 21st century

    National Research Council Canada - National Science Library

    Committee on Optical Science and Engineering; Commission on Physical Sciences, Mathematics, and Applications; Division on Engineering and Physical Sciences; National Research Council; National Academy of Sciences

    .... Harnessing Light surveys this multitude of applications, as well as the status of the optics industry and of research and education in optics, and identifies actions that could enhance the field's...

  14. 42 CFR 84.1133 - Harnesses; installation and construction; minimum requirements.

    Science.gov (United States)

    2010-10-01

    ... DEVICES Dust, Fume, and Mist; Pesticide; Paint Spray; Powered Air-Purifying High Efficiency Respirators and Combination Gas Masks § 84.1133 Harnesses; installation and construction; minimum requirements. (a...

  15. Børn og unge i opholdssteder - Hvordan har de det?

    DEFF Research Database (Denmark)

    Nissen, Poul; Mulbjerg, Steen

    Statusopgørelse vedrørende børn og unge i opholdssteder, som bl.a. viser, at  en tredjedel af drengene og halvdelen af pigerne har betydelige vanskeligheder i skolen og  forældre og pædagoger samstemmende rapporterer, at to tredjedele af drengene og fire femtedele af pigerne har kompromitteret so...

  16. Hvorfor skal Adam betale børnepenge, når Eva har bedraget ham?

    DEFF Research Database (Denmark)

    Petersen, Thomas Søbirk

    2016-01-01

    Juridisk abort bør være en mulighed for mænd, der er blevet bedraget, bestjålet eller som har brugt prævention, der ikke virker efter hensigten.......Juridisk abort bør være en mulighed for mænd, der er blevet bedraget, bestjålet eller som har brugt prævention, der ikke virker efter hensigten....

  17. Molecular mapping of a sunflower rust resistance gene from HAR6.

    Science.gov (United States)

    Bulos, Mariano; Ramos, María L; Altieri, Emiliano; Sala, Carlos A

    2013-03-01

    Sunflower rust, caused by Puccinia helianthi Schw., can result in significant yield losses in cultivated sunflower (Helianthus annuus L. var. macrocarpus Ckll.). HAR6 is a germplasm population resistant to most predominant rust races. The objectives of this study were to map the resistance factor present in HAR6 (R HAR6 ), and to provide and validate molecular tools for the identification of this gene for marker assisted selection purposes. Virulence reaction of seedlings for the F2 population and F2:3 families suggested that a single dominant gene confers rust resistance in HAR6-1, a selected rust resistance line from the original population. Genetic mapping with eight markers covered 97.4 cM of genetic distance on linkage group 13 of the sunflower consensus map. A co-dominant marker ZVG61 is the closest marker distal to R HAR6 at a genetic distance of 0.7 cM, while ORS581, a dominant marker linked in the coupling phase, is proximal to R HAR6 at a genetic distance of 1.5 cM. Validation of these markers was assessed by converting a susceptible line into a rust resistant isoline by means of marker assisted backcrossing. The application of these results to assist the breeding process and to design new strategies for rust control in sunflower is discussed.

  18. Hello, world: Harnessing social media for the Rosetta mission

    Science.gov (United States)

    Baldwin, Emily; Mignone, Claudia; O'Flaherty, Karen; Homfeld, Anne-Mareike; Bauer, Markus; McCaughrean, Mark

    2015-04-01

    The European Space Agency's (ESA) comet-chasing Rosetta mission was launched in 2004, before social media became a popular tool for mainstream communication. By harnessing a range of platforms for communicating the key messages of this unprecedented mission as it reached its destination ten years later, new audiences were reached and a global impact was achieved. Rosetta-specific social media accounts - @ESA_Rosetta on Twitter, the Rosetta Mission Facebook page and the rosettamission Instagram account - were developed during 2013/14 and used alongside the traditional reporting line of the main ESA website and the Rosetta blog to build awareness about the mission. Coordinated with ESA's existing social media channels (Flickr, YouTube, G+, Twitter, Facebook and Livestream) and with the support of ESA's country desks and Rosetta partner agency accounts (including @philae2014), information could be shared in a number of European languages, ensuring a wide reach across Europe - and the world. We discuss the roles of the various social media accounts in supporting and promoting the competitions and social media campaigns that were built around the key mission milestones of 2014: waking up from deep space hibernation (January), arriving at Comet 67P/Churyumov-Gerasimenko (August) and naming the landing site for Philae ahead of the landing event in November. We discuss the different approach to each channel, such as the first person twitter accounts, the dialogue with and between blog users, and the discussions held live via G+ Hangouts with leading scientists and spacecraft operators. We compare and contrast the audiences, the interaction we had with them and how challenges were overcome. We also use the science-fiction-meets-science-fact Ambition short movie, and its "undercover" dissemination on social media, as an example of how the profile of the Rosetta mission was raised in a unique way. By using a variety of social media platforms to target different audiences with

  19. An evaluation of backpack harness systems in non-neutral torso postures.

    Science.gov (United States)

    Southard, Stephanie A; Mirka, Gary A

    2007-09-01

    Much of the research on backpack design has been focused on spinal loading/biomechanics while the wearer is in a neutral/upright trunk posture, such as those employed by outdoor enthusiasts and schoolchildren. This research has led to some important harness design improvements that reduce trunk muscle exertions, fatigue and improve overall comfort. There are number of occupations, however, wherein workers wear back-mounted packs/devices (e.g. air tanks) while working in non-neutral trunk postures. The objective of the current study was to evaluate the effects of these non-neutral postures on biomechanical loading and then reconsider the backpack system design recommendations. Fifteen participants were asked to support a 18.2 kg load on their back while assuming static forward flexed postures of the torso (15 degrees , 30 degrees , 45 degrees , and 60 degrees of sagittal bend). The mass on the back was attached to the participant through two different harness mechanisms: a basic harness design (as seen on college student backpacks) and a more advanced design containing lateral stiffness rods and a weight-bearing hip belt (as seen on backpacks for hikers). While performing these static, posture maintenance tasks, the activation levels of the bilateral trapezius, erector spinae, and rectus abdominis were collected. Participants also provided subjective ratings of comfort. The results showed that there was a significant interaction between harness type and forward flexion angle for the trapezius and the erector spinae muscles. The normalized EMG for the trapezius muscles showed a 14% and 11% reduction in muscle activity at 15 degrees and 30 degrees , respectively, with the advanced design but these positive effects of the advanced design were not found at the greater flexion angles. Likewise the erector spinae muscles showed a 24% and 14% reduction in muscle activity at 15 degrees and 30 degrees , respectively, with the advanced design harness but these effects of the

  20. The use of a safety harness does not affect body sway during quiet standing.

    Science.gov (United States)

    Freitas, Sandra M S F; Prado, Janina M; Duarte, Marcos

    2005-03-01

    Postural sway during quiet standing reduces when somatosensorial information is provided by an active or passive "light touch" of different body parts with a surface. The contact of the safety harness with the body could induce a similar effect, leading to an undesirable side effect in the balance evaluation. This study investigated if a safety harness system, commonly used in balance studies, affects body sway during the balance evaluation. Healthy adults stood as quietly as possible for 60s in a comfortable position on a force plate. First, we performed an experiment on the light-touch effect with 10 subjects to determine the necessary sample size for the main investigation. Then, 60 subjects completed four tasks where the use of the safety harness and the visual information were manipulated. Area, root-mean square, speed, and frequency of the center of pressure displacement were analyzed. A light touch decreased postural sway on both visual conditions but there was no effect of the use of a safety harness on sway when quietly standing, independent of the visual information. Postural sway increased on both somatosensorial conditions when the visual information was not provided. This result shows that the safety harness does not interfere with the evaluation what is of major importance to methodological aspects of balance evaluation.

  1. Construction of an expression vector for Lactococcus lactis based on ...

    African Journals Online (AJOL)

    To construct an expression vector for Lactococcus lactis, the EmPMT fragment which contained the erythromycin resistance gene, P32 promoter, multiple cloning site (MCS) and terminator (T) was subcloned into the small cryptic plasmid pAR141. The resulting vector, designated as pAR1411, was found to be stably ...

  2. National Strategies to Harness Information Technology Seeking Transformation in Singapore, Finland, the Philippines, and South Africa

    CERN Document Server

    Knight, Peter

    2012-01-01

    The ability to harness Information Communication Technologies (ICTs) is increasingly at the heart of competitiveness and sustainable growth. As countries engage in an increasingly competitive global economy, they are trying to weave ICT into their development strategies, in the same way enterprises have learned to use ICT to transform their business models and strategies. This integration offers a new path to development that is responsive to the challenges of our times.  In National Strategies to Harness Information Technology, Nagy Hanna and Peter Knight provide a framework for assessing the opportunities, challenges, and prospects for “e-transformation” and for analyzing the options and innovations adopted to manage the e-transformation process.  They ask hard questions: what does it take to harness ICT to transform an economy?  Why some countries accelerate their development journey with ICT while others fail?  How did successful countries balance the need for strategic leadership with bottom up ...

  3. Use of isotopic tools to delimit areas of harnessing for drinking water supply - Final report

    International Nuclear Information System (INIS)

    Gourcy, L.; Petelet-Giraud, E.

    2011-03-01

    Within the frame of an action of the ONEMA-BRGM convention (Methodological approaches and tools for the protection of drinking water harnessing against diffuse pollutions), this study aims at developing a synthesis of isotopic geochemical tools to obtain the information required for the delimitation of harnessing supply areas. The report first describes the conventional tools: water molecule steady isotopes, radioactive isotopes, water dating tools, tools developed for another use, and artificial tracers. It presents the possible uses of natural and artificial tracers to determine parameters like flow directions, water residence duration, exchanges between aquifers and water sheet-river interactions. It gives an overview of knowledge on the use of isotopic methods to determine the origin of contaminants. It proposes a brief overview of a previous study of water sheets-rivers relationships. It finally discusses the use of geochemical and isotopic tools when delimiting supply areas for harnessing aimed at drinking water supply

  4. TRH-receptor mobility and function in intact and cholesterol-depleted plasma membrane of HEK293 cells stably expressing TRH-R-eGFP

    Czech Academy of Sciences Publication Activity Database

    Brejchová, Jana; Sýkora, Jan; Ostašov, Pavel; Merta, Ladislav; Roubalová, Lenka; Janáček, Jiří; Hof, Martin; Svoboda, Petr

    2015-01-01

    Roč. 1848, č. 3 (2015), s. 781-796 ISSN 0005-2736 R&D Projects: GA ČR(CZ) GAP207/12/0919 Institutional support: RVO:67985823 ; RVO:61388955 Keywords : cholesterol * TRH-R-eGFP mobility * FRAP * RICS * DPH fluorescence * G protein coupling Subject RIV: CE - Biochemistry; CF - Physical ; Theoretical Chemistry (UFCH-W) Impact factor: 3.687, year: 2015

  5. During Stably Suppressive Antiretroviral Therapy Integrated HIV-1 DNA Load in Peripheral Blood is Associated with the Frequency of CD8 Cells Expressing HLA-DR/DP/DQ

    Directory of Open Access Journals (Sweden)

    Alessandra Ruggiero

    2015-09-01

    Conclusions: The observed positive association between integrated HIV-1 DNA load and frequency of CD8+DR/DP/DQ+ cells indicates that a close correlation between HIV persistence and immune activation continues during consistently suppressive therapy. The inducers of the distinct activation profile warrant further investigation.

  6. Effectiveness of Mom-made Pavlik harness for maintaining reduction of the hip in DDH.

    Science.gov (United States)

    Lewsirirat, Supphamard; Piyapromdee, Urawit; Sathompanich, Suphasak

    2015-01-01

    Co-operation of the parent is a major factor for successful treatment of developmental dysplasia of the hip (DDH). The commercial Pavlik harness is too expensive and the parent may not understand or refuse its use. Regarding the function of the Pavlik harness and awareness of complications, a Mom-made Pavlik harness was developed. To evaluate the effectiveness of Mom-made Pavlik harness in maintaining the reduction ofDDH MATERIAL AND METHOD: Infants with Ortolani positive DDH, ages up to five months, treated at Maharat Nakhon Ratchasima Hospital between October 2003 and September 2012, were analyzed, retrospectively. Irreducible DDH, neuromuscular DDH and hyperlaxity syndrome were excluded. A Mom-made Pavlik harness was used to maintain reduction after the initial two weeks of a hip spica cast. The success of treatment was evaluated by ultrasound of the hip. The center edge (CE) angle was measured from AP film at age one and four years. There were 24 infants, 35 hips with 11 bilateral, seven left and six right sides DDH. Thirty-two hips were successfully reduced (91.4%). Three hips had recurrent dislocation. Two of these were further managed by adductor tenotomy and closed reduction under general anesthesia and hip spica cast, followed by hip abduction brace. Another was treated by open reduction and Salter's innominate osteotomy. The average CE angle at age one and four years was 16.5 and 25.9 degree, respectively. Treatment of the infants with DDH requires a long period of care not only in hospital but also at home. Therefore, it needs the diligence, intention and regularity by their parents to achieve a successful outcome. The Mom-made Pavlik harness represents the spirit of this resolution.

  7. Inquiry-based science education : - har naturfagsundervisningen i Danmark brug for det?

    DEFF Research Database (Denmark)

    Østergaard, Lars Domino; Sillasen, Martin Krabbe; Hagelskjær, Jens

    2010-01-01

    Inquiry-based science education (IBSE) er en internationalt afprøvet naturfagsdidaktisk metode der har til formål at øge elevernes interesse for og udbytte af naturfag. I artiklen redegøres der for metoden, der kan betegnes som en elevstyret problem- og undersøgelsesbaseret naturfagsundervisnings......Inquiry-based science education (IBSE) er en internationalt afprøvet naturfagsdidaktisk metode der har til formål at øge elevernes interesse for og udbytte af naturfag. I artiklen redegøres der for metoden, der kan betegnes som en elevstyret problem- og undersøgelsesbaseret...

  8. Science har fundet løsning på CURIS-registrering

    DEFF Research Database (Denmark)

    Mørch, Frede

    2012-01-01

    Fakultetsbibliotekar Frede Mørch fortæller her om den løsning, som Science har indført for at sikre at alle forskningspublikationer bliver registreret i databasen, CURIS, uden at forskerne skal spilde tid på det.......Fakultetsbibliotekar Frede Mørch fortæller her om den løsning, som Science har indført for at sikre at alle forskningspublikationer bliver registreret i databasen, CURIS, uden at forskerne skal spilde tid på det....

  9. Conversion of Human Fibroblasts to Stably Self-Renewing Neural Stem Cells with a Single Zinc-Finger Transcription Factor

    Directory of Open Access Journals (Sweden)

    Ebrahim Shahbazi

    2016-04-01

    Full Text Available Direct conversion of somatic cells into neural stem cells (NSCs by defined factors holds great promise for mechanistic studies, drug screening, and potential cell therapies for different neurodegenerative diseases. Here, we report that a single zinc-finger transcription factor, Zfp521, is sufficient for direct conversion of human fibroblasts into long-term self-renewable and multipotent NSCs. In vitro, Zfp521-induced NSCs maintained their characteristics in the absence of exogenous factor expression and exhibited morphological, molecular, developmental, and functional properties that were similar to control NSCs. In addition, the single-seeded induced NSCs were able to form NSC colonies with efficiency comparable with control NSCs and expressed NSC markers. The converted cells were capable of surviving, migrating, and attaining neural phenotypes after transplantation into neonatal mouse and adult rat brains, without forming tumors. Moreover, the Zfp521-induced NSCs predominantly expressed rostral genes. Our results suggest a facilitated approach for establishing human NSCs through Zfp521-driven conversion of fibroblasts.

  10. Construction of a high-EGFR expression cell line and its biological ...

    African Journals Online (AJOL)

    Targeted screening of EGFR compounds has become one of the medical research focuses for tumor therapy. A431, which naturally expresses high levels of EGFR, was compared with the stably high expressing EGFR cell line HEK293. Flow cytometry was used to analyze cell growth and Western blot was used to ...

  11. Comparison of plant-based expression platforms for the heterologous production of geraniol

    NARCIS (Netherlands)

    Vasilev, N.; Schmitz, C.; Dong, L.; Ritala, A.; Imseng, N.; Hakkinen, S.T.; Krol, van der A.R.; Eibl, R.; Oksman-Caldentey, K.M.; Bouwmeester, H.J.; Fischer, R.; Schillberg, S.

    2014-01-01

    We compared the ability of different plant-based expression platforms to produce geraniol, a key metabolite in the monoterpenoid branch of the terpenoid indole alkaloid biosynthesis pathway. A geraniol synthase gene isolated from Valeriana officinalis (VoGES) was stably expressed in different

  12. The expression of GFP under the control of fibroin promotor in ...

    Indian Academy of Sciences (India)

    Unknown

    The fibroin promoter can stably express foreign gene in lepidopteran cells. Total RNA was extracted from the gland of silkworm, Antheraea pernyi and the transcription initiation site of fibroin gene of A. pernyi was identi- fied by RNA ligase mediated rapid amplification of cDNA ends (RLM-RACE). The expression vector ...

  13. The expression of GFP under the control of fibroin promotor in ...

    Indian Academy of Sciences (India)

    The fibroin promoter can stably express foreign gene in lepidopteran cells. Total RNA was extracted from the gland of silkworm, Antheraea pernyi and the transcription initiation site of fibroin gene of A. pernyi was identified by RNA ligase mediated rapid amplification of cDNA ends (RLM-RACE). The expression vector ...

  14. Fem sociale medier har »uindskrænket ret« til at misbruge alt dit indhold

    DEFF Research Database (Denmark)

    Nielsen, Jøren Ullits Olai

    2016-01-01

    Ved du, hvor meget kontrol du har over dit indhold på Facebook, YouTube, Snapchat, Instagram og Twitter? Her giver en juraforsker dig det forkromede overblik.......Ved du, hvor meget kontrol du har over dit indhold på Facebook, YouTube, Snapchat, Instagram og Twitter? Her giver en juraforsker dig det forkromede overblik....

  15. Harnessing the power of big data: infusing the scientific method with machine learning to transform ecology

    Science.gov (United States)

    Most efforts to harness the power of big data for ecology and environmental sciences focus on data and metadata sharing, standardization, and accuracy. However, many scientists have not accepted the data deluge as an integral part of their research because the current scientific method is not scalab...

  16. Universities plan revolution in research; Four institutes will harness top brains

    CERN Multimedia

    Buie, E

    2003-01-01

    "Scotland is on the brink of revolutionising its approach to university research by creating four institutions that would harness the country's top brains to work together in specific areas. The move to create pan-academic institutes in physics, life-sciences, economics and creative arts has won the backing of the principals of every higher education institution in Scotland..." (1 page).

  17. How to Move in a Jostling Crowd The Art of Harnessing Random ...

    Indian Academy of Sciences (India)

    Home; Journals; Resonance – Journal of Science Education; Volume 1; Issue 10. How to Move in a Jostling Crowd The Art of Harnessing Random Motions. G S Ranganath. Research News Volume 1 Issue 10 October 1996 pp 84-86. Fulltext. Click here to view fulltext PDF. Permanent link:

  18. Secondary geochemical reactions in hydrothermal energy harnessing; Geochemische Folgereaktionen bei der hydrogeothermalen Energiegewinnung

    Energy Technology Data Exchange (ETDEWEB)

    Kuehn, M.

    1996-12-01

    This thesis is in six parts: description of geothermal energy harnessing, analytics, geochemical-thermodynamic modellings, experimental investigations, discussion of results, summary. (HW) [Deutsch] Die vorliegende Dissertationsschrift gliedert sich in 6 Teile: - Beschreibung der geothermischen Energienutzung - Analytik - geochemische-thermodynamische Modellierungen - experimentelle Untersuchungen - Diskussion der Ergebnisse - Zusammenfassung. (HW)

  19. Improving Students' Educational Experience by Harnessing Digital Technology: elgg in the ODL Environment

    Science.gov (United States)

    Tung, Lai Cheng

    2013-01-01

    Given the rising popularity of both open and distance learning (ODL) and social networking tools, it seems logical to merge and harness these two popular technologies with the goal of improving student educational experience. The integration seems to hold tremendous promise for the open and distance learning mode. To reduce the gap in the…

  20. Taso Mähar : maali ümber ehitatud maja / Gitte Hint

    Index Scriptorium Estoniae

    Hint, Gitte

    2003-01-01

    220-ruutmeetrise ühekorde eramu sisekujundus. Arhitekt Emil Urbel. Sisearhitekt Taso Mähar, tema kommentaarid. Köögimööbel on T. Mähari projekteeritud. Elutoas mööblifirma T&T Mang pehme mööbel, seinal Lemming Nageli maal "Punane joon all - Burn Out". Ill.: plaan, 17 värv. vaadet

  1. Harnessing ICT Potential: The Adoption and Analysis of ICT Systems for Enhancing the Student Learning Experience

    Science.gov (United States)

    Dawson, Shane; Heathcote, Liz; Poole, Gary

    2010-01-01

    Purpose: This paper aims to examine how effective higher education institutions have been in harnessing the data capture mechanisms from their student information systems, learning management systems and communication tools for improving the student learning experience and informing practitioners of the achievement of specific learning outcomes.…

  2. Warming Up to Trade? Harnessing International Trade to Support Climate Change Objectives

    OpenAIRE

    World Bank

    2007-01-01

    This study on harnessing international trade to support climate change objectives assesses the following: 1) What are the main policy prescriptions for reducing greenhouse gases that are employed by OECD countries and how do they impact the competitiveness of their energy-intensive industries? 2) On account of the impact on competitiveness, is there is leakage of energy intensive industrie...

  3. Pressure distribution under three different types of harnesses used for guide dogs.

    Science.gov (United States)

    Peham, C; Limbeck, S; Galla, K; Bockstahler, B

    2013-12-01

    The aim of this study was to evaluate the pressure distribution under three different types of harnesses used for guide dogs (designated H1, H2 and H3). The dogs (n = 8) led a trainer through a course including a range of exercises (straight line, curve left, curve right, upstairs and downstairs). All dogs were clinically sound and showed no sign of lameness. The pressures beneath the harnesses were determined by sensor strips and related to the gait. In all harnesses, the highest pressures were found in the right sternal region (H1 2.02 ± 0.6N/cm(2); H2 1.76 ± 0.4N/cm(2); H3 1.14 ± 0.5 N/cm(2)). In all other regions, the pressures were in the range of 0-1.32 N/cm(2). The right and left sternal regions were almost constantly loaded. Contrary to previous assumptions, the back regions had minimal loading. This investigation demonstrated that there were significant differences among the harnesses. Copyright © 2013 Elsevier Ltd. All rights reserved.

  4. Harnessing the plurality of actor frames in social-ecological systems : Ecological sanitation in Bolivia

    NARCIS (Netherlands)

    Eelderink, M.; Vervoort, J.; Snel, D.; de Castro, F.

    2017-01-01

    This article uses a case study on ecological sanitation as a basis for lessons on identifying and harnessing the plurality of actor frames in social-ecological systems, thereby moving beyond the advocacy positions often taken by implementing NGOs. The study aimed to explore how perspectives between

  5. The Facebook-in-Action: Challenging, Harnessing and Enhancing Students Class Assignments and Projects

    Science.gov (United States)

    Saifudin, Adam Mohd; Yacob, Aizan; Saad, Rohaizah

    2016-01-01

    Issues of universities students harnessing and capitalizing the usage of Facebook for their own learning capabilities and effective thinking is always the focus of education scholars, in assessing the quality class assignments and projects produced by them. Therefore, Facebook is now becoming unbearable influence since the internet activation in…

  6. Harnessing eugenol as a substrate for production of aromatic compounds with recombinant strains of Amycolatopsis sp. HR167.

    Science.gov (United States)

    Overhage, Jörg; Steinbüchel, Alexander; Priefert, Horst

    2006-09-18

    To harness eugenol as cheap substrate for the biotechnological production of aromatic compounds, the vanillyl alcohol oxidase gene (vaoA) from Penicillium simplicissimum CBS 170.90 was cloned in an expression vector suitable for Gram-positive bacteria and expressed in the vanillin-tolerant Gram-positive strain Amycolatopsis sp. HR167. Recombinant strains harboring hybrid plasmid pRLE6SKvaom exhibited a specific vanillyl alcohol oxidase activity of 1.1U/g protein. Moreover, this strain had gained the ability to grow on eugenol as sole carbon source. The intermediates coniferyl alcohol, coniferyl aldehyde, ferulic acid, guajacol, and vanillic acid were detected as excreted compounds during growth on eugenol, whereas vanillin could only be detected in trace amounts. Resting cells of Amycolatopsis sp. HR167 (pRLE6SKvaom) produced coniferyl alcohol from eugenol with a maximum conversion rate of about 2.3 mmol/h/l of culture, and a maximum coniferyl alcohol concentration of 4.7 g/1 was obtained after 16 h biotransformation without further optimization. Beside coniferyl alcohol, traces of coniferyl aldehyde and ferulic acid were also detected.

  7. Water-resistant, monodispersed and stably luminescent CsPbBr3/CsPb2Br5 core-shell-like structure lead halide perovskite nanocrystals

    Science.gov (United States)

    Qiao, Bo; Song, Pengjie; Cao, Jingyue; Zhao, Suling; Shen, Zhaohui; Gao, Di; Liang, Zhiqin; Xu, Zheng; Song, Dandan; Xu, Xurong

    2017-11-01

    Lead halide perovskite materials are thriving in optoelectronic applications due to their excellent properties, while their instability due to the fact that they are easily hydrolyzed is still a bottleneck for their potential application. In this work, water-resistant, monodispersed and stably luminescent cesium lead bromine perovskite nanocrystals coated with CsPb2Br5 were obtained using a modified non-stoichiometric solution-phase method. CsPb2Br5 2D layers were coated on the surface of CsPbBr3 nanocrystals and formed a core-shell-like structure in the synthetic processes. The stability of the luminescence of the CsPbBr3 nanocrystals in water and ethanol atmosphere was greatly enhanced by the photoluminescence-inactive CsPb2Br5 coating with a wide bandgap. The water-stable enhanced nanocrystals are suitable for long-term stable optoelectronic applications in the atmosphere.

  8. Failed Pavlik harness treatment for DDH as a risk factor for avascular necrosis.

    Science.gov (United States)

    Tiruveedhula, Madhu; Reading, Isabel C; Clarke, Nicholas M P

    2015-03-01

    Avascular necrosis (AVN) of the femoral head is an irreversible complication seen in the treatment of developmental dysplasia of hip (DDH) with the Pavlik harness. Its incidence is reported to be low after successful reduction of the hip but high if the hip is not concentrically relocated. We aim to investigate its incidence after failed Pavlik harness treatment. We prospectively followed up a group of children who failed Pavlik harness treatment for DDH treated at our institution by the senior author between 1988 and 2001 and compared their rates of AVN with a group of children who presented late and hence were treated surgically. AVN was graded as described by Kalamchi and MacEwen and only grade 2 to 4 AVN was considered significant and included in the analysis. Thirty-seven hips were included in the failed Pavlik group (group 1) and 86 hips in the no Pavlik group (group 2). Ten hips in group 1 developed AVN (27%), whereas only 7 hips in group 2 (8%) developed AVN; the odds of developing AVN after failed Pavlik treatment was 4.7 (95% confidence interval, 1.3-14.1) (P=0.009) with a relative risk of 3.32 (range, 1.37 to 8.05). There was no statistically significant association observed with duration of splintage and severity of AVN (Spearman's correlation, -0.46; P=0.18). However, there was a positive correlation noted with age at presentation and severity of AVN. Therefore, we advise close monitoring of hips in the Pavlik harness and discontinue its use if the hips are not reduced within 3 weeks. Level III.

  9. The relevance of netnography to the harness of Romanian health care electronic word-of-mouth.

    Science.gov (United States)

    Bratucu, R; Gheorghe, I R; Radu, A; Purcarea, V L

    2014-09-15

    Nowadays, consumers use the computer mediated communication to make purchase decisions on a large variety of products and services. Since health care services are archetypal by nature, consumers in this field are one of the most encountered users of electronic word-of-mouth. The objective of this paper is to explain and support the necessity of adopting a different qualitative method when electronic word of mouth is harnessed on health care dedicated forums, that is, netnography.

  10. Følelser, fiktion – og Facebook - har erobret den politiske arena

    DEFF Research Database (Denmark)

    Aagaard, Peter

    2016-01-01

    Er der ét træk de nye politiske bevægelser – fra Trump til Alternativet – har til fælles, er det evnen til at fortælle historier, vælgerne kan koble sig på som medfortællere. Historier om håb og frelse, fornedrelse og fortabelse. Det bringer drømmene og magien tilbage i politik. Derfor vinder de...

  11. Enhanced Harnessing of the Graviola Bioactive Components Using a Neoteric Sonication Cum Microwave Coadjuvant Extraction Protocol

    OpenAIRE

    Se Chul Chun; Shang Xiaomin; Vimala Anthonydhason; Hyejin Jung; Shimels Tilahun Belachew; Judy Gopal; Diby Paul

    2018-01-01

    Graviola is one of the most accomplished natural anticancer therapists gaining popularity in recent times. Harnessing the full benefit from tapping all of its rich bioactive reservoirs is absolutely worthy and mandatory. It is in this regard that a well optimized extraction methodology gains paramount importance. In case of Graviola, no sophistication in terms of extraction methods is reported. A neoteric sonication cum microwave combined extraction technology was introduced that maximized th...

  12. A Comparative Study of Harnessing Cattle - An Important Chapter in the History of European Ethnology

    Czech Academy of Sciences Publication Activity Database

    Woitsch, Jiří

    2017-01-01

    Roč. 38, č. 2017 (2017), s. 59-89 ISSN 1335-4116 R&D Projects: GA ČR GA15-03754S Institutional support: RVO:68378076 Keywords : harnessing cattle * European ethnology * history of ethnology Subject RIV: AC - Archeology, Anthropology, Ethnology OBOR OECD: History (history of science and technology to be 6.3, history of specific sciences to be under the respective headings)

  13. Application Layer Definition and Analyses of Controller Area Network Bus for Wire Harness Assembly Machine

    OpenAIRE

    Guo, Hui; Jiang, Ying

    2006-01-01

    With the feature of multi-master bus access, nondestructive contention-based arbitration and flexible configuration, Controller Area Network (CAN) bus is applied into the control system of Wire Harness Assembly Machine (WHAM). To accomplish desired goal, the specific features of the CAN bus is analyzed by compared with other field buses and the functional performances in the CAN bus system of WHAM is discussed. Then the application layer planning of CAN bus for dynamic priority is presented. ...

  14. Harnessing microbial gene pools to remediate persistent organic pollutants using genetically modified plants--a viable technology?

    Science.gov (United States)

    Rylott, Elizabeth L; Johnston, Emily J; Bruce, Neil C

    2015-11-01

    It has been 14 years since the international community came together to legislate the Stockholm Convention on Persistent Organic Pollutants (POPs), restricting the production and use of specific chemicals that were found to be environmentally stable, often bioaccumulating, with long-term toxic effects. Efforts are continuing to remove these pollutants from the environment. While incineration and chemical treatment can be successful, these methods require the removal of tonnes of soil, at high cost, and are damaging to soil structure and microbial communities. The engineering of plants for in situ POP remediation has had highly promising results, and could be a more environmentally-friendly alternative. This review discusses the characterization of POP-degrading bacterial pathways, and how the genes responsible have been harnessed using genetic modification (GM) to introduce these same abilities into plants. Recent advances in multi-gene cloning, genome editing technologies and expression in monocot species are accelerating progress with remediation-applicable species. Examples include plants developed to degrade 2,4,6-trinitrotoluene (TNT), hexahydro-1,3,5-trinitro-1,3,5-triazine (RDX), trichloroethylene (TCE), and polychlorinated biphenyls (PCBs). However, the costs and timescales needed to gain regulatory approval, along with continued public opposition, are considerable. The benefits and challenges in this rapidly developing and promising field are discussed. © The Author 2015. Published by Oxford University Press on behalf of the Society for Experimental Biology. All rights reserved. For permissions, please email: journals.permissions@oup.com.

  15. Riding Horse Harness (Based on Materials from the Chulkovo Burial Ground

    Directory of Open Access Journals (Sweden)

    Grishakov Valeriy V.

    2013-12-01

    Full Text Available The items of riding horse harness (bits with psalia, stirrups, buckles, decorations of bridles are analyzed in the article on the basis of the materials from the Chulkovo (Murom burial ground site of the 8th-10th centuries, located on right bank of the Oka river. It has been established that the use of horse harness items by the Muroma culture population, which had left the Chulkovo burial ground, is in full correspondence with the general traditions of the Finno-Ugric peoples of the Volga river region characteristic of the 8th-10th centuries. The shapes of the bits correspond to both the steppe (bits with S-shaped and straight psalia and the local traditions (bits with two and three moving rings. A tendency to uniformity is observed in the tradition of stirrups production. The main differences of the Muroma burial grounds from the neighboring Mordovian ones consist in the presence of horse burials (not known among the Mordovians, and, on the other hand, in a practically complete lack of horse harness in the funeral set of human burials. It can be assumed that the importance of cavalry with the Muromа was lower than with the Mordovians who were living next to the steppe world.

  16. Mapping the Sea Floor of the Historic Area Remediation Site (HARS) Offshore of New York City

    Science.gov (United States)

    Butman, Bradford

    2002-01-01

    The area offshore of New York City has been used for the disposal of dredged material for over a century. The area has also been used for the disposal of other materials such as acid waste, industrial waste, municipal sewage sludge, cellar dirt, and wood. Between 1976 and 1995, the New York Bight Dredged Material Disposal Site, also known as the Mud Dump Site (MDS), received on average about 6 million cubic yards of dredged material annually. In September 1997 the MDS was closed as a disposal site, and it and the surrounding area were designated as the Historic Area Remediation Site (HARS). The sea floor of the HARS, approximately 9 square nautical miles in area, currently is being remediated by placing a minimum 1-m-thick cap of clean dredged material on top of the surficial sediments that are contaminated from previous disposal of dredged and other materials. The U.S. Geological Survey (USGS) is working cooperatively with the U.S. Army Corps of Engineers (USACE) to map the sea floor geology of the HARS and changes in the characteristics of the surficial sediments over time.

  17. The Belem Framework for Action: Harnessing the Power and Potential of Adult Learning and Education for a Viable Future

    Science.gov (United States)

    Adult Learning, 2012

    2012-01-01

    This article presents the Belem Framework for Action. This framework focuses on harnessing the power and potential of adult learning and education for a viable future. This framework begins with a preamble on adult education and towards lifelong learning.

  18. Technical note: A simple back-mounted harness for grazing dairy cows to facilitate the sulfur hexafluoride tracer gas technique.

    Science.gov (United States)

    van Wyngaard, Josef D V; Meeske, Robin; Erasmus, Lourens J

    2018-03-01

    We describe here a cattle harness to attach a gas collection vessel to facilitate the sulfur hexafluoride (SF 6 ) tracer gas technique. The harness consists of 2 major components: (1) a lightweight, robust body fabricated from an equine surcingle or lunge roller with padded thoracic trapezius pressure points, a bespoke shaping shaft for spine support, and adjustable buckles on both sides; and (2) an elastic flank-strap to prevent the harness from dislodging. The spine support consists of stainless steel laminated with carbon fiber. This support minimizes the contact area with the animal's skin, relieves the spine area of pressure, and creates free flow of ambient air below the platform, reducing sweat accumulation and hence preventing skin lesions. The harness weighs approximately 1.2 kg, allows for attachment of 2 gas collection vessels (animal and background sample), and is cost effective. Copyright © 2018 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.

  19. Double-Stranded Linear Duck Hepatitis B Virus (DHBV) Stably Integrates at a Higher Frequency than Wild-Type DHBV in LMH Chicken Hepatoma Cells

    Science.gov (United States)

    Gong, Shih S.; Jensen, Anne D.; Chang, C. J.; Rogler, Charles E.

    1999-01-01

    Integration of hepadnavirus DNAs into host chromosomes can have oncogenic consequences. Analysis of host-viral DNA junctions of DHBV identified the terminally duplicated r region of the viral genome as a hotspot for integration. Since the r region is present on the 5′ and 3′ ends of double-stranded linear (DSL) hepadnavirus DNAs, these molecules have been implicated as integration precursors. We have produced a LMH chicken hepatoma cell line (LMH 66-1 DSL) which replicates exclusively DSL duck hepatitis B virus (DHBV) DNA. To test whether linear DHBV DNAs integrate more frequently than the wild type open circular DHBV DNAs, we have characterized the integration frequency in LMH 66-1 DSL cells by using a subcloning approach. This approach revealed that 83% of the LMH 66-1 DSL subclones contained new integrations, compared to only 16% of subclones from LMH-D2 cells replicating wild-type open circular DHBV DNA. Also, a higher percentage of the LMH 66-1 DSL subclones contained two or more new integrations. Mathematical analysis suggests that the DSL DHBV DNAs integrated stably once every three generations during subcloning whereas wild-type DHBV integrated only once every four to five generations. Cloning and sequencing of new integrations confirmed the r region as a preferred integration site for linear DHBV DNA molecules. One DHBV integrant was associated with a small deletion of chromosomal DNA, and another DHBV integrant occurred in a telomeric repeat sequence. PMID:9882355

  20. Analytical and numerical study of natural convection in a stably stratified fluid along vertical plates and cylinders with temporally-periodic surface temperature variations

    International Nuclear Information System (INIS)

    Shapiro, A.; Fedorovich, E.

    2005-01-01

    This paper describes one-dimensional (parallel) laminar and transitional regimes of natural convection in a viscous stably stratified fluid due to temporally-periodic variations in the surface temperature of infinite vertical plates and cylinders. Analytical solutions are obtained for the periodic laminar regime for arbitrary values of stratification, Prandtl number and forcing frequency. The solutions for plates and cylinders are qualitatively similar and show that (i) the flows are composed of two waves that decay exponentially with distance from the surface; a fast long wave and a slow short wave, (ii) for forcing frequencies less than the natural frequency, both waves propagate away from the surface, while (iii) for forcing frequencies less than this natural frequency, the short wave propagates away from the surface while the long wave propagates toward the surface. The analytical results are complemented, for the plate problem, with three-dimensional numerical simulations of flows that start from rest and are suddenly subjected to a periodic thermal forcing. The numerical results depict the transient (start-up) stage of the laminar flow and the approach to the periodicity, and confirm that the analytical solutions provide the appropriate description of the periodic regime for the laminar convection case. Preliminary numerical data are presented for transition from the laminar to turbulent convection. (authors)

  1. Microfibrillated cellulose sheets coating oxygen-permeable PDMS membranes induce rat hepatocytes 3D aggregation into stably-attached 3D hemispheroids.

    Science.gov (United States)

    Evenou, Fanny; Couderc, Sandrine; Kim, Beomjoon; Fujii, Teruo; Sakai, Yasuyuki

    2011-01-01

    Here we report the use of natural, chemically-unmodified, microfibrillated cellulose (MFC) as a matrix for hepatocyte culture. We developed an original cell-culture design composed of a thin 3D-microstructured fibrous substrate consisting of a MFC sheet coating a highly O(2)-permeable polydimethylsiloxane (PDMS) membrane. The MFC-coated PDMS membranes were obtained according to a simple process where cellulose fibres were deposited from an aqueous suspension on the PDMS surfaces and the films were dried under mild conditions. To enable oxygen diffusion through the membranes, they were assembled on bottomless frames ('O(2)+' condition). Rat hepatocytes primary-cultured on such MFC-PDMS membranes quickly organized themselves into large hemispherical 3D aggregates which were tightly anchored to the MFC sheets. In contrast, hepatocytes cultured on smooth PDMS membranes in the O(2)+ system (O(2)+, PDMS) organized into unstable 2D monolayers which easily detached from the surfaces. Hepatocyte 3D cultures obtained on MFC-PDMS membranes exhibited higher liver-specific functions over a 2-week culture period, as assessed by both the higher albumin secretion and urea synthesis rate. The MFC-PDMS membranes appear suitable for obtaining stably-attached and functional hepatocyte 3D cultures and appear interesting for drug/chemical screenings in a microplate format, but also for microfluidic applications.

  2. Enhanced Harnessing of the Graviola Bioactive Components Using a Neoteric Sonication Cum Microwave Coadjuvant Extraction Protocol

    Directory of Open Access Journals (Sweden)

    Se Chul Chun

    2018-02-01

    Full Text Available Graviola is one of the most accomplished natural anticancer therapists gaining popularity in recent times. Harnessing the full benefit from tapping all of its rich bioactive reservoirs is absolutely worthy and mandatory. It is in this regard that a well optimized extraction methodology gains paramount importance. In case of Graviola, no sophistication in terms of extraction methods is reported. A neoteric sonication cum microwave combined extraction technology was introduced that maximized the extraction process and minimized (7 min the extraction time. The extraction efficiency was validated based on the significant enrichment of bioactive ingredients in Graviola extracts following the sonication cum microwave combined protocol.

  3. Deaf Workers in Restaurant, Retail, and Hospitality Sector Employment: Harnessing Research to Promote Advocacy.

    Science.gov (United States)

    Stokar, Hayley

    2017-01-01

    A quarter-century after the passage of the Americans with Disabilities Act (ADA, 1990 ), workplace accommodation is still a struggle for deaf employees and their managers. Many challenges are the result of communication barriers that can be overcome through much needed-although often absent-advocacy and training. This article highlights the literature on the employment of deaf individuals in the United States service industries of food service, retail, and hospitality conducted from 2000 to 2016. Exploring dimensions of both hiring and active workplace accommodation, suggestions are made for how social work advocates can harness information and strengthen their approaches for educating managers and supporting workers.

  4. Harnessing the beneficial properties of adipogenic microbes for improving human health

    DEFF Research Database (Denmark)

    Dhurandhar, N V; Geurts, L; Atkinson, R L

    2013-01-01

    protection from obesity-induced metabolic comorbidities. To this end, the 10th Stock conference examined new approaches to improve metabolic comorbidities independent of weight loss. In particular, human adenovirus 36 (Ad36) and specific gut microbes were examined for their potential to influence lipid...... and glucose homeostasis in animals and humans. While these microbes possess some undesirable properties, research has identified attributes of adenovirus Ad36 and gut microbes that may be selectively harnessed to improve metabolic profile without the obligatory weight loss. Furthermore, identifying the host...

  5. DEAD-box helicase DDX27 regulates 3′ end formation of ribosomal 47S RNA and stably associates with the PeBoW-complex

    Energy Technology Data Exchange (ETDEWEB)

    Kellner, Markus; Rohrmoser, Michaela [Department of Molecular Epigenetics, Helmholtz Center Munich, Center for Integrated Protein Science Munich (CIPSM), Marchioninistr. 25, Munich 81377 (Germany); Forné, Ignasi [Adolf Butenandt Institute, Ludwig Maximilians University of Munich, Center for Integrated Protein Science Munich (CIPSM), Schillerstr. 44, Munich 80336 (Germany); Voss, Kirsten; Burger, Kaspar; Mühl, Bastian; Gruber-Eber, Anita [Department of Molecular Epigenetics, Helmholtz Center Munich, Center for Integrated Protein Science Munich (CIPSM), Marchioninistr. 25, Munich 81377 (Germany); Kremmer, Elisabeth [Institute of Molecular Immunology, Helmholtz Center Munich, Marchioninistr. 25, Munich 81377 (Germany); Imhof, Axel [Adolf Butenandt Institute, Ludwig Maximilians University of Munich, Center for Integrated Protein Science Munich (CIPSM), Schillerstr. 44, Munich 80336 (Germany); Eick, Dirk, E-mail: eick@helmholtz-muenchen.de [Department of Molecular Epigenetics, Helmholtz Center Munich, Center for Integrated Protein Science Munich (CIPSM), Marchioninistr. 25, Munich 81377 (Germany)

    2015-05-15

    PeBoW, a trimeric complex consisting of pescadillo (Pes1), block of proliferation (Bop1), and the WD repeat protein 12 (WDR12), is essential for processing and maturation of mammalian 5.8S and 28S ribosomal RNAs. Applying a mass spectrometric analysis, we identified the DEAD-box helicase DDX27 as stably associated factor of the PeBoW-complex. DDX27 interacts with the PeBoW-complex via an evolutionary conserved F×F motif in the N-terminal domain and is recruited to the nucleolus via its basic C-terminal domain. This recruitment is RNA-dependent and occurs independently of the PeBoW-complex. Interestingly, knockdown of DDX27, but not of Pes1, induces the accumulation of an extended form of the primary 47S rRNA. We conclude that DDX27 can interact specifically with the Pes1 and Bop1 but fulfils critical function(s) for proper 3′ end formation of 47S rRNA independently of the PeBoW-complex. - Highlights: • DEAD-box helicase DDX27 is a new constituent of the PeBoW-complex. • The N-terminal F×F motif of DDX27 interacts with the PeBoW components Pes1 and Bop1. • Nucleolar anchoring of DDX27 via its basic C-terminal domain is RNA dependent. • Knockdown of DDX27 induces a specific defect in 3′ end formation of 47S rRNA.

  6. Ecoenvironment characteristics of subsidence lakes in coal mine areas and ways to harness them

    International Nuclear Information System (INIS)

    Bian Zhengfu; Zhang Guoliang; Lin Jiacong

    1994-01-01

    With the increase in coal output, the environment in coal mine areas, especially land destruction, has become increasingly worse. A main form of land destruction is subsidence lakes, which have disrupted people's lives and coal production and which change the ecological system of mine areas into water-land type or aquatic system. Two conditions forming subsidence lakes are surface subsidence and high phreatic level. The ratio of waterlogging in China's eastern coal mine areas is higher than that of other areas because of high phreatic level, and eastern mine areas are agriculture-based at the same time. It is a fact that the structure and function of an ecological system after mining is different from that before mining. It is not a good way to resume the original structure and function of the ecological system. Therefore, before harnessing subsidence lakes, it is necessary that their ecoenvironment characteristics be investigated thoroughly, then new structure and function of ecology affected by coal mining and ways to harness subsidence lakes are posed. These ways include: subsidence lakes used as fly ash, coal waste and other wastes deposit, land reclamation by raising the shallow land with digging the deep, recreating new aquatic ecosystem, etc

  7. Fatigue life prediction of a cable harness in an industrial robot using dynamic simulation

    Energy Technology Data Exchange (ETDEWEB)

    Yoon, Ji Won; Park, Tae Won [Ajou University, Suwon (Korea, Republic of); Yim, Hong Jae [Kookmin University, Seoul (Korea, Republic of)

    2008-03-15

    The cable which transfers the signal and power in an industrial robot has a problem of fatigue fracture like steel components. Since the cable is very flexible compared to other components of the system, it is difficult to estimate its motion numerically. Some studies have been done on a large deformation problem, especially in a cable, and a few attempts have been made to apply the absolute nodal coordinate formulation (ANCF), which can simulate a large deformation. Only researches about the fatigue life of structural cables or comparative studies of FEM and ANCF simulations can be found. This paper presents a method to simulate the behavior of the cable harness using the ANCF and to predict the fatigue life while computing the strain time history of the point of interest. Rigid body dynamics is applied for the robot system, while ANCF is used for the cable harness. The simulation is performed by using the dynamic analysis process. The material property of the cable is obtained by a test. A simplified model is prepared. With these data, the behavior of the cable is simulated and the fatigue life is predicted

  8. Effects of harness transmitters on behavior and reproduction of wild mallards

    Science.gov (United States)

    Pietz, Pamela J.; Krapu, Gary L.; Greenwood, Raymond J.; Lokemoen, John T.

    1993-01-01

    Radio telemetry has been an important research tool in waterfowl studies for >20 years, yet little effort has been made to evaluate potential effects of transmitters on the birds that carry them. As part of a 4-year mallard (Anas platyrhynchos) study in the prairie pothole region of North Dakota and Minnesota, we compared radio-marked and unmarked female mallards in terms of percent time observed feeding, resting, and preening; nest initiation date; and clutch size and egg volume. Radio-marked females carried a 23-g back-mounted transmitter attached with a 2-loop harness (Dwyer 1972). On average, radio-marked females tended to feed less, rest and preen more, initiate nests later, and lay smaller clutches and eggs than unmarked females. Thus, behavioral and reproductive data from ducks marked with back-mounted harness-attached transmitters may be biased. We recommend that new designs of radio packages be field tested and caution that effects may be masked under extreme environmental conditions.

  9. Tectonothermal evolution of the Triassic flysch in the Bayan Har Orogen, Tibetan plateau

    Science.gov (United States)

    Wang, Hejing; Rahn, Meinert; Zhou, Jian

    2018-01-01

    The Bayan Har Orogen comprises a major part of the "Qingzang-Dianxi fold region" in western China. It preserves important information of the tectono-thermal evolution covering the time span from the closure of the Paleo-Tethys Ocean up to the formation of the Himalayas. Low temperature metamorphic indicators, such as mineral assemblages, illite "crystallinity" (IC), chlorite "crystallinity" (CC), illite polytype, b-cell dimension of K-white micas, geothermometry of selected minerals were analyzed. The values of Kübler index (KI) of the Triassic flysch in the Bayan Har Orogen range from 0.23-1.63°Δ2θ while Árkai index (ÁI) in a range of 0.21-0.60°Δ2θ. Iso-thermal zones mapped with KI describe a pair of anchizones and an anchiregion within the Bayan Har Orogen: the "Giant Yushu Anchizone" in the southwest (extending > 750 km long and 100 km wide), the "Zaling-Eling-Lakes Anchizone" in the center (about 150 km long and 40 km wide) and the "Xing-Tong-Zhe Anchiregion" in the northeast (covering an area of roughly 60,000 km2). They are separated by diagenetic zones. Peak metamorphic conditions are estimated around 280-330 °C and a low to intermediate (N. New Hampshire) pressure type. A slight change with increasing then decreasing pressure was observed from SW to NE. The relationship between anchimetamorphic pattern of Triassic flysch and large-scale folds and faults indicates syn- to post structural metamorphism. Compression at the end of the Triassic, induced by the interaction of the Tarim, North China and Indian blocks caused the closure of the Paleo-Tethys Ocean and led to the folding of the Triassic flysch within the Paleo-Tethys Ocean basin. Anchimetamorphism may have been caused by crustal thickening of > 10 km due to an accretionary wedge setting and a temperature increase in those rocks due to burial. Such a regional metamorphic pattern would provide important information for reconstruction of palaeotectonic-palaeogeograph and the evolutionary history

  10. Trump har droppet fløjlshandskerne, nu da han er slået tilbage til start

    DEFF Research Database (Denmark)

    Hendricks, Vincent Fella

    2016-01-01

    Donald Trump vandt primærvalget ved hjælp af personangreb. I præsidentvalgkampens slutspurt har han nu taget den strategi op igen, selvom alle dømte den ude – og det er måske slet ikke så dumt, for det er det, Trump kan......Donald Trump vandt primærvalget ved hjælp af personangreb. I præsidentvalgkampens slutspurt har han nu taget den strategi op igen, selvom alle dømte den ude – og det er måske slet ikke så dumt, for det er det, Trump kan...

  11. Housekeeping genes for quantitative expression studies in the three-spined stickleback Gasterosteus aculeatus

    OpenAIRE

    Hibbeler, S.; Scharsack, J.P.; Becker, S.

    2008-01-01

    Abstract Background During the last years the quantification of immune response under immunological challenges, e.g. parasitation, has been a major focus of research. In this context, the expression of immune response genes in teleost fish has been surveyed for scientific and commercial purposes. Despite the fact that it was shown in teleostei and other taxa that the gene for beta-actin is not the most stably expressed housekeeping gene (HKG), depending on the tissue and experimental treatmen...

  12. ZigBee Test Harness: An Innovative Tool for ZigBee Node Testing

    Directory of Open Access Journals (Sweden)

    Andrea Ranalli

    2010-12-01

    Full Text Available The document describes an innovative tool called Test Harness, developed by Telecom Italia S.p.A (TIT.MI. The purpose of this software is to tests any ZigBee node compliant to the ZigBee public application profile specifications. The tool can be used to debug a device before its certification process, a step that needs to be done within the ZigBee Alliance in order to commercialize a product with the ZigBee logo. Typical testing include finding malformed packets format, wrong field values, checks the correct behavior of the node when receiving or sending messages, etc… The tool tries to speed up the activities of Test Houses, helping them to automate a test plan. Finally, the tool supports also a distribute mode testing, allowing companies to run remote testing and reduce budget cost (flights, accommodation, etc....

  13. Wind turbine blades for harnessing energy from Malaysian low speed wind - manufacturing technique

    International Nuclear Information System (INIS)

    Abas Abd Wahab; Azmin Shakrine

    2000-01-01

    Blades for wind turbine to harness energy in the Malaysia low speed winds have been designed. During wind tunnel testing, wind turbine model using this type of blades has cut in speed of 1.5 m/s and turned at 450 rpm at 4 m/s wind. The blades, due to their critical dimensions of 1.2 m length, 5 cm thickness, tapered and 15 degree twist, were difficult to produce especially in large number. Several production methods have been studied but for economical mass production, fibreglass blades using CNC cutting mould were chosen. The blade and mould designs and the manufacturing processes are briefly outlined in this paper. (Author)

  14. How is knowledge co-produced in dialogue through the harnessing of “difference”?

    DEFF Research Database (Denmark)

    Phillips, Louise Jane; Olesen, Birgitte Ravn; Nielsen, Jørgen Lerche

    This panel is about the “dialogic turn” in the production and communication of knowledge, a general societal tendency in which practices of co-creation are widespread across diverse fields of social practice as a means of generating knowledge, often with a view to practice change. Practices of co....... In the dialogic turn, communication is configured as dialogue in which multiple social actors co-produce knowledge collaboratively across multiple knowledge forms and knowledge interests. Participants in dialogue, it is claimed, are empowered as self-directed co-learners, co-researchers or co...... in dialogue. By harnessing difference as a transformative force, it is claimed, dialogue can generate knowledge across differences, including differences of organizational and professional position, theoretical perspective, gender, ethnicity, class and so on. But how exactly is knowledge co...

  15. Harnessing Vehicle Automation for Public Mobility -- An Overview of Ongoing Efforts

    Energy Technology Data Exchange (ETDEWEB)

    Young, Stanley E.

    2015-11-05

    This presentation takes a look at the efforts to harness automated vehicle technology for public transport. The European CityMobil2 is the leading demonstration project in which automated shuttles were, or are planned to be, demonstrated in several cities and regions. The presentation provides a brief overview of the demonstrations at Oristano, Italy (July 2014), LaRochelle, France (Dec 2014), Lausanne, Switzerland (Apr 2015), Vantaa, Finland (July 2015), and Trikala, Greece (Sept 2015). In addition to technology exposition, the objectives included generating a legal framework for operation in each location and gaging the reaction of the public to unmanned shuttles, both of which were successfully achieved. Several such demonstrations are planned throughout the world, including efforts in North America in conjunction with the GoMentum Station in California. These early demonstration with low-speed automated shuttles provide a glimpse of the possible with a fully automated fleet of driverless vehicle providing a public transit service.

  16. Let Our Powers Combine! Harnessing NASA's Earth Observatory Natural Event Tracker (EONET) in Worldview

    Science.gov (United States)

    Wong, Min Minnie; Ward, Kevin; Boller, Ryan; Gunnoe, Taylor; Baynes, Kathleen; King, Benjamin

    2016-01-01

    Constellations of NASA Earth Observing System (EOS) satellites orbit the earth to collect images and data about the planet in near real-time. Within hours of satellite overpass, you can discover where the latest wildfires, severe storms, volcanic eruptions, and dust and haze events are occurring using NASA's Worldview web application. By harnessing a repository of curated natural event metadata from NASA Earth Observatory's Natural Event Tracker (EONET), Worldview has moved natural event discovery to the forefront and allows users to select events-of-interest from a curated list, zooms to the area, and adds the most relevant imagery layers for that type of natural event. This poster will highlight NASA Worldviews new natural event feed functionality.

  17. The sweetest pill to swallow: how patient neurobiology can be harnessed to maximise placebo effects.

    Science.gov (United States)

    Jubb, Jayne; Bensing, Jozien M

    2013-12-01

    The burgeoning interest in placebo effects over the last 10-15 years has fallen into two main research areas: elucidation of the neurobiological mechanisms recruited following placebo administration, and investigations into the situations and contexts in which placebo effects are evoked. There has been little attention focused on bridging these two i.e. how to actively translate and apply these neurobiological mechanisms into daily clinical practice in a responsible way. This article addresses this gap, first through a narrative review of the last 15 years of neuroscience findings with special attention focussed on the elucidation of the neurotransmitters, pathways and mechanisms involved in placebo effects, and secondly, at how these psycho(neuro)biological effects could be harnessed in medical care. Copyright © 2013 Elsevier Ltd. All rights reserved.

  18. Harnessing type I and type III CRISPR-Cas systems for genome editing

    DEFF Research Database (Denmark)

    Li, Yingjun; Pan, Saifu; Zhang, Yan

    2016-01-01

    CRISPR-Cas (clustered regularly interspaced short palindromic repeats-CRISPR-associated) systems are widespread in archaea and bacteria, and research on their molecular mechanisms has led to the development of genome-editing techniques based on a few Type II systems. However, there has not been any...... report on harnessing a Type I or Type III system for genome editing. Here, a method was developed to repurpose both CRISPR-Cas systems for genetic manipulation in Sulfolobus islandicus, a thermophilic archaeon. A novel type of genome-editing plasmid (pGE) was constructed, carrying an artificial mini-CRISPR...... and selectively retained as transformants. Using this strategy, different types of mutation were generated, including deletion, insertion and point mutations. We envision this method is readily applicable to different bacteria and archaea that carry an active CRISPR-Cas system of DNA interference provided...

  19. Antiviral Goes Viral: Harnessing CRISPR/Cas9 to Combat Viruses in Humans.

    Science.gov (United States)

    Soppe, Jasper Adriaan; Lebbink, Robert Jan

    2017-10-01

    The clustered regularly interspaced short palindromic repeats (CRISPR) and CRISPR-associated (Cas) systems are RNA-guided sequence-specific prokaryotic antiviral immune systems. In prokaryotes, small RNA molecules guide Cas effector endonucleases to invading foreign genetic elements in a sequence-dependent manner, resulting in DNA cleavage by the endonuclease upon target binding. A rewired CRISPR/Cas9 system can be used for targeted and precise genome editing in eukaryotic cells. CRISPR/Cas has also been harnessed to target human pathogenic viruses as a potential new antiviral strategy. Here, we review recent CRISPR/Cas9-based approaches to combat specific human viruses in humans and discuss challenges that need to be overcome before CRISPR/Cas9 may be used in the clinic as an antiviral strategy. Copyright © 2017 Elsevier Ltd. All rights reserved.

  20. Genetic suppressors of the Lotus japonicus har1-1 hypernodulation phenotype

    DEFF Research Database (Denmark)

    Murray, Jeremy; Karas, Bogumil; Ross, Loretta

    2006-01-01

    approach of genetic mapping, targeting induced local lesions in genomics, and sequencing, all non-nodulating mutant lines were characterized and shown to represent new alleles of at least nine independent symbiotic loci. The class of mutants with reduced nodulation capacity was of particular interest...... mutant lines carried more subtle symbiotic phenotypes, either forming white ineffective nodules or showing reduced nodulation capacity. When challenged with Glomus intraradices, 18 of the 61 suppressor lines were unable to establish a symbiosis with this arbuscular mycorrhiza fungus. Using a combined...... because some of them may specify novel plant functions that regulate nodule development in L. japonicus. To facilitate mapping of the latter class of mutants, an introgression line, in which the har1-1 allele was introduced into a polymorphic background of L. japonicus ecotype MG20, was constructed...

  1. Harnessing big data for estimating the energy consumption and driving range of electric vehicles

    DEFF Research Database (Denmark)

    Fetene, Gebeyehu Manie; Kaplan, Sigal; Mabit, Stefan Lindhard

    2017-01-01

    to conventional ones. Moreover, the results of the analysis show that driving speed, acceleration and temperature have non-linear effects on the ECR, while season and precipitation level have a strong linear effect. The econometric model of the ECR of BEVs suggests that the optimal driving speed is between 45......Analyzing the factors that affect the energy efficiency of vehicles is crucial to the overall improvement of the environmental efficiency of the transport sector, one of the top polluting sectors at the global level. This study analyses the energy consumption rate (ECR) and driving range of battery...... electric vehicles (BEVs) and provides insight into the factors that affect their energy consumption by harnessing big data from real-world driving. The analysis relied on four data sources: (i) driving patterns collected from 741 drivers over a two-year period; (ii) drivers’ characteristics; (iii) road...

  2. Opportunities for high aspect ratio micro-electro-magnetic-mechanical systems (HAR-MEMMS) at Lawrence Berkeley Laboratory

    Energy Technology Data Exchange (ETDEWEB)

    Hunter, S. [ed.

    1993-10-01

    This report contains viewgraphs on the following topics: Opportunities for HAR-MEMMS at LBL; Industrial Needs and Opportunities; Deep Etch X-ray Lithography; MEMS Activities at BSAC; DNA Amplification with Microfabricated Reaction Chamber; Electrochemistry Research at LBL; MEMS Activities at LLNL; Space Microsensors and Microinstruments; The Advanced Light Source; Institute for Micromaching; IBM MEMS Interests; and Technology Transfer Opportunities at LBL.

  3. Evaluating the High Asia Reanalysis (HAR) using Gauge-based and Satellite Precipitation Data over High Mountain Asia

    Science.gov (United States)

    Pangaluru, K.; Velicogna, I.; Ciraci, E.; Mohajerani, Y.

    2017-12-01

    The Indus, Ganges and Brahmaputra (IGB) basins supply water for both domestic and agricultural demands, the latter of which is the mainstay of Indian economy. Here, we use high-resolution Asia Refined Analysis (HAR) rainfall datasets to study the spatial and temporal behavior of rainfall over the mountainous areas of the Indus, Ganges and Brahmaputra (IGB) over the period from 2001 to 2014. The validation of High Asia Refined Analysis (HAR) precipitation data is carried out with observational (GPCP, CRU and CPC) and satellite (TRMM_3B43) datasets for the period. We find that the relative differences between the HAR model and the satellite and gauge-based datasets varies between -9% and 67% for the seasonal mean and between 1% and 26% for the annual mean for all basins. The correlation between the HAR model and the observational datasets lies between 0.5 and 0.9 for all seasons. Spatial variations and monthly magnitudes of gridded precipitation trends are calculated by using the Mann-Kendall (MK) test and the Thei-Sen approach (TSA) respectively. We found significant positive trends precipitation grids over the IGB basins in the annual and monsoon season time frames, as opposed to winter and falls seasons.

  4. Opportunities for high aspect ratio micro-electro-magnetic-mechanical systems (HAR-MEMMS) at Lawrence Berkeley Laboratory

    International Nuclear Information System (INIS)

    Hunter, S.

    1993-10-01

    This report contains viewgraphs on the following topics: Opportunities for HAR-MEMMS at LBL; Industrial Needs and Opportunities; Deep Etch X-ray Lithography; MEMS Activities at BSAC; DNA Amplification with Microfabricated Reaction Chamber; Electrochemistry Research at LBL; MEMS Activities at LLNL; Space Microsensors and Microinstruments; The Advanced Light Source; Institute for Micromaching; IBM MEMS Interests; and Technology Transfer Opportunities at LBL

  5. Ipsilateral Scapular Cutaneous Anchor System : An alternative for the harness in body-powered upper-limb prostheses

    NARCIS (Netherlands)

    Hichert, M.; Plettenburg, D.H.

    2017-01-01

    Background: Body-powered prosthesis users frequently complain about the poor cosmesis and comfort of the traditional shoulder harness. The Ipsilateral Scapular Cutaneous Anchor System offers an alternative, but it remains unclear to what extent it affects the perception and control of cable

  6. Some estimated effects of the planned harnessing of the Ounasjoki river on reindeer husbandry

    Directory of Open Access Journals (Sweden)

    Pirkko Nieminen

    1983-05-01

    Full Text Available The harnessing of waterways for electrical power has caused permanent pasture losses and prevented the free movement of the reindeer herds in Finland. Many great changes occurred after construction of the two large artificial lakes of Lokka and Porttipahta (total 630 km2 in the Lappi reindeer herders association in the 1960s. The planned harnessing of the Ounasjoki river consists of 10 power plants and 2 big and 12 smaller artificial lakes (total 270 km2. The plan will have effects on the income of 1070 owners in 7 reindeer herders associations. The losses in reindeer husbandry estimated by three different methods were maximally 476, 2824 and 9900 reindeer (value of meat production 0.12 to 2.5 million FIM/year, 64 jobs and various herding buildings (value 3.9 million FIM. Two new reindeer farmes would become unusuable (0.5 million FIM and in addition hay production from seasonally flooded fields (approx. 25 000 - 30 000 FIM/year would be lost. The building of new forces in the reindeer herders association areas of Ounasjoki river would require 6.2 million FIM.Ounasjoen rakentamissuunnitelman mahdolliset vaikutukset porotalouteen.Abstract in Finnsish / Yhteenveto: Vesistojen valjastaminen såhkontuottoon on tuhonnut porolaitumia ja vaikcuttanut porojen vapaata liikkumista Suomessa. Tasta on hyvånå esimerkkinå Lokan ja Portipahdan tekoaltaiden (yhteenså 630 km2 rakentaminen Lapin paliskunnassa 1960-luvulta alkacn. Ounasjoen rakentamissuunnitelma kåsittåå 10 voimalaitosta ja 2 isoa ja 12 picnempåå tekoallasta (yhteenså 270 km2. Rakennussuunnitelma vaikuttaa 1070 poronomistajan talouteen 7 cri paliskunnassa. Kolmella eri mcnetelmallå laskien jouduttaisiin enimmillåån våhentåmåån 476, 2824 ja 9900 lukuporoa (lihantuotto 0,12-2,5 milj. mk/vuosi sekå menetettåisim 64 ympårivuotista tyopaikkaa ja kocttaisiin useita eri rakennevahinkoja (arvoltaan noin 3,9 milj. mk. Kaksi uutta porotilaa jåisi kåyttokclvottomiksi (0,5 milj. mk ja

  7. Transition through co-optation: Harnessing carbon democracy for clean energy

    Science.gov (United States)

    Meng, Kathryn-Louise

    This dissertation explores barriers to a clean energy transition in the United States. Clean energy is demonstrably viable, yet the pace of clean energy adoption in the U.S. is slow, particularly given the immediate threat of global climate change. The purpose of this dissertation is to examine the factors inhibiting a domestic energy transition and to propose pragmatic approaches to catalyzing a transition. The first article examines the current political-economic and socio-technical energy landscape in the U.S. Fossil fuels are central to the functioning of the American economy. Given this centrality, constellations of power have been constructed around the reliable and affordable access of fossil fuels. The fossil fuel energy regime is comprised of: political-economic networks with vested interests in continued fossil fuel reliance, and fixed infrastructure that is minimally compatible with distributed generation. A transition to clean energy threatens the profitability of fossil fuel regime actors. Harnessing structural critiques from political ecology and process and function-oriented socio-technical systems frameworks, I present a multi-level approach to identifying pragmatic means to catalyzing an energy transition. High-level solutions confront the existing structure, mid-level solutions harness synergy with the existing structure, and low-level solutions lie outside of the energy system or foster the TIS. This is exemplified using a case study of solar development in Massachusetts. Article two presents a case study of the clean energy technological innovation system (TIS) in Massachusetts. I examine the actors and institutions that support cleantech development. Further, I scrutinize the actors and institutions that help sustain the TIS support system. The concept of a catalyst is presented; a catalyst is an actor that serves to propel TIS functions. Catalysts are critical to facilitating anchoring. Strategic corporate partners are identified as powerful

  8. In vivo photoacoustic imaging of tyrosinase expressing tumours in mice

    Science.gov (United States)

    Laufer, Jan; Jathoul, Amit; Johnson, Peter; Zhang, Edward; Lythgoe, Mark; Pedley, R. Barbara; Pule, Martin; Beard, Paul

    2012-02-01

    Two human tumour cell lines (K562, 293T) were stably transfected to achieve the genetic expression of tyrosinase, which is involved in the production of the pigment eumelanin. The cells were injected subcutaneously into nude mice to form tumour xenografts, which were imaged over a period of up to 26 days using an all-optical photoacoustic imaging system. 3D photoacoustic images of the tumours and the surrounding vasculature were acquired at excitation wavelengths ranging from 600nm to 770nm. The images showed tumour growth and continued tyrosinase expression over the full 26 day duration of the study. These findings were confirmed by histological analysis of excised tumour samples.

  9. Construction of pEgr-hTRAIL expression vector induced by irradiaiton and its apoptosis in tumor cells in vitro

    International Nuclear Information System (INIS)

    Piao Chunji; Tian Mei; Yang Wei; Li Xiuyi; Liu Linlin

    2005-01-01

    Objective: To construct the radiation-inducible expression vector pEgr-hTRAIL containing human TNF related apoptosis inducing ligand (hTRAIL) gene and study its expression and function of inducing apoptosis in A549 human lung adenocarcinoma cells. Methods: Expression vector pEgr-hTRAIL was constructed with DNA recombinant technique. pEgr-hTRAIL plasmids were packaged with lipofectin to transfect into A549 cells in vitro. Stably transfected cell line A549-shTRAIL was selected through G418. The expression of hTRAIL gene was detected by RT-PCR. The early stage apoptosis of A549 cells was detected by Annexin-V-FITC apoptosis detecting kit. Results: Expression vector pEgr-hTRAIL was constructed correctly by identification with restriction enzyme digestion. The expression of hTRAIL mRNA in stably transfected cell line A549-shTRAIL was increased significantly. The percentage of apoptotic A549-shTRAIL was increased significantly; it was 1.8 times as much as A549 cells (P<0.05). Conclusion: Expression vector pEgr-hTRAIL is constructed successfully, which can increase the apoptosis of the stably transfected cell line A549-shTRAIL significantly. (authors)

  10. Feline and canine coronaviruses are released from the basolateral side of polarized epithelial LLC-PK1 cells expressing the recombinant feline aminopeptidase-N cDNA

    NARCIS (Netherlands)

    Rossen, J W; Kouame, J; Goedheer, A J; Vennema, H; Rottier, P J

    2001-01-01

    In this study feline (FECV and FIPV) and canine (CCoV) coronavirus entry into and release from polarized porcine epithelial LLC-PK1 cells, stably expressing the recombinant feline aminopeptidase-N cDNA, were investigated. Virus entry appeared to occur preferentially through the apical membrane,

  11. Newly constructed stable reporter cell lines for mechanistic studies on electrophile-responsive element-mediated gene expression reveal a role for flavonoid planarity

    NARCIS (Netherlands)

    Boerboom, A.M.J.F.; Vermeulen, M.; Woude, H. van der; Bremer, B.I.; Lee-Hilz, Y.Y.; Kampman, E.; Bladeren, P.J. van; Rietjens, I.M.C.M.; Aarts, J.M.M.J.G.

    2006-01-01

    The electrophile-responsive element (EpRE) is a transcriptional enhancer involved in cancer-chemoprotective gene expression modulation by certain food components. Two stably transfected luciferase reporter cell lines were developed, EpRE(hNQO1)-LUX and EpRE(mGST-Ya)-LUX, based on EpRE sequences from

  12. Validation of a rapid yeast estrogen bioassay, based on the expression of green fluorescent protein, for the screening of estrogenic activity in calf urine

    NARCIS (Netherlands)

    Bovee, T.F.H.; Heskamp, H.H.; Hamers, A.R.M.; Hoogenboom, L.A.P.; Nielen, M.W.F.

    2005-01-01

    Previously we described the construction and properties of a rapid yeast bioassay stably expressing human estrogen receptor a (hERa) and yeast enhanced green fluorescent protein (yEGFP) in response to estrogens. In the present study, this yeast estrogen assay was validated as a qualitative screening

  13. Newly constructed stable reporter cell lines for mechanistic studies on electrophile-responsive element-mediated gene expression reveal a role for flavonoid planarity.

    NARCIS (Netherlands)

    Boerboom, A.M.A.; Vermeulen, M.; Woude, H. van der; Bremer, B.I.; Lee-Hilz, Y.Y.; Kampman, E.; Bladeren, P.J. van; Rietjens, I.M.C.M.; Aarts, J.

    2006-01-01

    The electrophile-responsive element (EpRE) is a transcriptional enhancer involved in cancer-chemoprotective gene expression modulation by certain food components. Two stably transfected luciferase reporter cell lines were developed, EpRE(hNQO1)-LUX and EpRE(mGST-Ya)-LUX, based on EpRE sequences from

  14. Harnessing the immune system through programmed death-1 blockade in the management of Hodgkin lymphoma

    Directory of Open Access Journals (Sweden)

    Oncale MB

    2017-02-01

    Full Text Available Melody B Oncale, Hossein Maymani, Loretta J Nastoupil Department of Lymphoma and Myeloma, Division of Cancer Medicine, The University of Texas MD Anderson Cancer Center, Houston, TX, USA Abstract: Immunotherapy is a rapidly evolving therapeutic option in the treatment of lymphoma. Neoplastic cells evade immune recognition through the programmed death (PD-1/PD-ligand immune checkpoint pathway. Several novel agents have been developed to restore the immune system’s ability to recognize and destroy cancer cells. Nivolumab and pembrolizumab are two anti-PD-1 antibodies that have demonstrated success in the treatment of refractory Hodgkin lymphoma. Harnessing the immune system’s ability to target neoplastic cells, ideally without the use of cytotoxic chemotherapeutic agents, is one way in which these novel agents are changing the therapeutic landscape in the treatment of lymphomas. Here, we review the emerging data regarding checkpoint inhibitors in the management of Hodgkin lymphoma, the unique adverse effects encountered with the use of these agents, and a practical approach to the management of these adverse effects. Additionally, we discuss upcoming trials that will further assess the promising future developments of checkpoint inhibition in the treatment of not only Hodgkin lymphoma but also other B cell lymphomas and myeloma. These agents offer immense promise of a future where many lymphomas can be treated without the toxic effects of chemotherapeutic agents. Keywords: Hodgkin lymphoma, programmed death-1, nivolumab, pembrolizumab, lymphoma

  15. Periodic trends governing the interactions between impurity atoms [H-Ar] and (alpha)-U

    Energy Technology Data Exchange (ETDEWEB)

    Taylor, Christopher David [Los Alamos National Laboratory

    2008-01-01

    The binding energies, geometries, charges and electronic structures of a series of impurity atoms [H-Ar] interacting with the {alpha}-U lattice in various configurations were assessed by means of density functional theory calculations. Periodic trends governing the binding energy were highlighted and related to the electronic properties of the impurity atoms, with some consideration given to the band-structure of {alpha}-U. The strongest bound impurity atoms include [C, N, O] and [Si, P, S]. The general trends in the binding energy can be reproduced by a simple parameterisation in terms of the electronegativity (charge-transfer) and covalent radius (elasticity theory) of the impurity atom. The strongest bound atoms deviate from this model, due to their ability to bind with an optimum mixture of covalency and ionicity. This last point is evidenced by the partial overlap of the impurity atom p-band with the hybrid d-/f-band of {alpha}-U. It is expected that the trends and general behaviour reported in this work can be extended to the interactions of impurity atoms with other metallic systems.

  16. Harnessing surface-bound enzymatic reactions to organize microcapsules in solution.

    Science.gov (United States)

    Shklyaev, Oleg E; Shum, Henry; Sen, Ayusman; Balazs, Anna C

    2016-03-01

    By developing new computational models, we examine how enzymatic reactions on an underlying surface can be harnessed to direct the motion and organization of reagent-laden microcapsules in a fluid-filled microchannel. In the presence of appropriate reagents, surface-bound enzymes can act as pumps, which drive large-scale fluid flows. When the reagents diffuse through the capsules' porous shells, they can react with enzymatic sites on the bottom surface. The ensuing reaction generates fluid density variations, which result in fluid flows. These flows carry the suspended microcapsules and drive them to aggregate into "colonies" on and near the enzyme-covered sites. This aggregation continues until the reagent has been depleted and the convection stops. We show that the shape of the assembled colonies can be tailored by patterning the distribution of enzymes on the surface. This fundamental physicochemical mechanism could have played a role in the self-organization of early biological cells (protocells) and can be used to regulate the autonomous motion and targeted delivery of microcarriers in microfluidic devices.

  17. Harnessing Wind Power in Moving Reference Frames with Application to Vehicles

    Science.gov (United States)

    Goushcha, Oleg; Felicissimo, Robert; Danesh-Yazdi, Amir; Andreopoulos, Yiannis

    2017-11-01

    The extraction of wind power from unique configurations embedded in moving vehicles by using micro-turbine devices has been investigated. In such moving environments, the specific power of the air motion is much greater and less intermittent than in stationary wind turbines anchored to the ground in open atmospheric conditions. In a translational frame of reference, the rate of work done by the drag force acting on the wind harnessing device due the relative motion of air should be taken into account in the overall performance evaluation through an energy balance. A device with a venting tube has been tested that connects a high-pressure stagnating flow region in the front of the vehicle with a low-pressure region at its rear. Our analysis identified two key areas to focus on for potentially significant rewards: (1) Vehicles with high energy conversion efficiency which require a high mass flow rate through the venting duct, and (2) low efficiency vehicles with wakes, which will be globally affected by the introduction of the venting duct device in a manner that reduces their drag so that there is a net gain in power generation.

  18. Harnessing Geothermal Energy from CO2 Enhanced Oil Recovery (EOR) Operations

    Science.gov (United States)

    Saar, M. O.; Randolph, J. B.

    2012-12-01

    Recent geotechnical research shows that geothermal heat can be efficiently mined by circulating CO2 through naturally permeable, porous rock formations. This method, called CO2 Plume Geothermal (CPG), targets the same geologic reservoirs that are suitable for deep saline aquifer CO2 sequestration or enhanced oil recovery (EOR). While previous investigations have focused on CO2-based heat mining from saline aquifers, here we present new research that is primarily concerned with EOR reservoirs, specifically those using a CO2 flood. EOR operations provide excellent opportunities for economically-favorable geothermal energy recovery, assuming subsurface temperatures are sufficient, because the majority of costly infrastructure (i.e., wells) is in place. Moreover, the subsurface characteristics that make a site suitable for hydrocarbon recovery -- at least moderate reservoir permeability and porosity, and a low-permeability capping feature -- help ensure that fluid can be circulated for heat extraction and that CO2 will be contained. However, heat extraction from the CO2 + water/brine + hydrocarbon EOR production stream is challenging, requiring fluid separation and multiple binary and/or direct power systems (depending on site-specific fluid composition and conditions). We discuss several scenarios, encompassing multiple power system configurations, for harnessing geothermal energy from CO2 EOR operations. In addition, we present preliminary numerical modeling results for net power production from such EOR operations -- accounting for wide variation in produced fluid temperature, pressure, and composition -- and consider the economic implications of power sales for EOR sites.

  19. Harnessing cell-to-cell variations to probe bacterial structure and biophysics

    Science.gov (United States)

    Cass, Julie A.

    Advances in microscopy and biotechnology have given us novel insights into cellular biology and physics. While bacteria were long considered to be relatively unstructured, the development of fluorescence microscopy techniques, and spatially and temporally resolved high-throughput quantitative studies, have uncovered that the bacterial cell is highly organized, and its structure rigorously maintained. In this thesis I will describe our gateTool software, designed to harness cell-to-cell variations to probe bacterial structure, and discuss two exciting aspects of structure that we have employed gateTool to investigate: (i) chromosome organization and the cellular mechanisms for controlling DNA dynamics, and (ii) the study of cell wall synthesis, and how the genes in the synthesis pathway impact cellular shape. In the first project, we develop a spatial and temporal mapping of cell-cycle-dependent chromosomal organization, and use this quantitative map to discover that chromosomal loci segregate from midcell with universal dynamics. In the second project, I describe preliminary time- lapse and snapshot imaging analysis suggesting phentoypical coherence across peptidoglycan synthesis pathways.

  20. Harnessing real world data from wearables and self-monitoring devices: feasibility, confounders and ethical considerations

    Directory of Open Access Journals (Sweden)

    Uttam Barick

    2016-07-01

    Full Text Available The increasing usage of smart phones has compelled mobile technology to become a universal part of everyday life. From wearable gadgets to sophisticated implantable medical devices, the advent of mobile technology has completely transformed the healthcare delivery scenario. Self-report measures enabled by mobile technology are increasingly becoming a more time and cost efficient method of assessing real world health outcomes. But, amidst all the optimism, there are concerns also on adopting this technology as regulations and ethical considerations on privacy legislations of end users are unclear. In general, the healthcare industry functions on some stringent regulations and compliances to ensure the safety and protection of patient information. A couple of the most common regulations are Health Insurance Portability Accountability Act (HIPPA and Health Information Technology for Economic and Clinical Health (HITECH. To harness the true potential of mobile technology to empower stakeholders and provide them a common platform which seamlessly integrates healthcare delivery and research, it is imperative that challenges and drawbacks in the sphere are identified and addressed. In this age of information and technology, no stones should be left unturned to ensure that the human race has access to the best healthcare services without an intrusion into his/her confidentiality. This article is an overview of the role of tracking and self-monitoring devices in data collection for real world evidence/observational studies in context to feasibility, confounders and ethical considerations.

  1. Deep Unconventional Geothermal Resources: a major opportunity to harness new sources of sustainable energy

    Energy Technology Data Exchange (ETDEWEB)

    Fridleifsson, G.O.; Albertsson, A.; Stefansson, B.; Gunnlaugsson, E.; Adalsteinsson, H.

    2007-07-01

    The Iceland Deep Drilling Project (IDDP) is a long-term program to improve the efficiency and economics of geothermal energy by harnessing Deep Unconventional Geothermal Resources (DUGR). Its aim is to produce electricity from natural supercritical hydrous fluids from drillable depths. Producing supercritical fluids will require drilling wells and sampling fluids and rocks to depths of 3.5 to 5 km, and at temperatures of 450-600{sup o}C. The long-term plan is to drill and test a series of such deep boreholes in Iceland at the Krafla, the Hengill, and the Reykjanes high temperature geothermal systems. Beneath these three developed drill fields temperatures should exceed 550-650{sup o}C, and the occurrence of frequent seismic activity below 5 km, indicates that the rocks are brittle and therefore likely to be permeable. Modeling indicates that if the wellhead enthalpy is to exceed that of conventionally produced geothermal steam, the reservoir temperature must be higher than 450{sup o}C. A deep well producing 0.67 m3/sec steam ({approx}2400 m3/h) from a reservoir with a temperature significantly above 450{sup o}C could yield enough high-enthalpy steam to generate 40-50 MW of electric power. This exceeds by an order of magnitude the power typically obtained from conventional geothermal wells. (auth)

  2. Harnessing coherence of area decomposition and semantic shared spaces for task allocation in a robotic fleet

    Directory of Open Access Journals (Sweden)

    Domagoj Drenjanac

    2014-08-01

    Full Text Available Task allocation is a fundamental problem in multi-robot systems where heterogeneous robots cooperate to perform a complex mission. A general requirement in a task allocation algorithm is to find an optimal set of robots to execute a certain task. This paper presents the work that harnesses an area decomposition algorithm, and a space-based middleware to facilitate task allocation process in unstructured and dynamic environments. To reduce spatial interference between robots, area decomposition algorithm divides a working area into cells which are then dynamically assigned to robots. In addition, coordination and collaboration among distributed robots are realized through a space-based middleware. For this purpose, the space-based middleware is extended with a semantic model of robot capabilities to improve task selection in terms of flexibility, scalability, and reduced communication overhead during task allocation. In this way a framework which exploits the synergy of area decomposition and semantically enriched space-based approach is created. We conducted performance tests in a specific precision agriculture use case focusing on the utilization of a robotic fleet for weed control introduced in the European Project RHEA – Robot Fleets for Highly Effective Agriculture and Forestry Management.

  3. MRI Evaluation of Spinal Length and Vertebral Body Angle During Loading with a Spinal Compression Harness

    Science.gov (United States)

    Campbell, James A.; Hargens, Alan R.; Murthy, G.; Ballard, R. E.; Watenpaugh, D. E.; Hargens, Alan, R.; Sanchez, E.; Yang, C.; Mitsui, I.; Schwandt, D.; hide

    1998-01-01

    Weight bearing by the spinal column during upright posture often plays a role in the common problem of low back pain. Therefore, we developed a non-ferromagnetic spinal compression harness to enable MRI investigations of the spinal column during axial loading. Human subjects were fitted with a Nest and a footplate which were connected by adjustable straps to an analog load cell. MRI scans of human subjects (5 males and 1 female with age range of 27-53 yrs) during loaded and unloaded conditions were accomplished with a 1.5 Tesla GE Signa scanner. Studies of two subjects undergoing sequentially increasing spinal loads revealed significant decreases (r(sup 2) = 0.852) in spinal length between T4 and L5 culminating in a 1.5 to 2% length decrease during loading with 75% body weight. Sagittal vertebral body angles of four subjects placed under a constant 50% body weight load for one hour demonstrated increased lordotic and kyphotic curvatures. In the lumbar spine, the L2 vertebral body experienced the greatest angular change (-3 deg. to -5 deg.) in most subjects while in the thoracic spine, T4 angles increased from the unloaded state by +2 deg. to +9 deg. Overall, our studies demonstrate: 1) a progressive, although surprisingly small, decrease in spinal length with increasing load and 2) relatively large changes in spinal column angulation with 50% body weight.

  4. Harnessing structural darkness in the visible and infrared wavelengths for a new source of light

    KAUST Repository

    Huang, Jianfeng

    2015-10-19

    Engineering broadband light absorbers is crucial to many applications, including energy-harvesting devices and optical interconnects. The performances of an ideal absorber are that of a black body, a dark material that absorbs radiation at all angles and polarizations. Despite advances in micrometre-thick films, the absorbers available to date are still far from an ideal black body. Here, we describe a disordered nanostructured material that shows an almost ideal black-body absorption of 98-99% between 400 and 1,400 nm that is insensitive to the angle and polarization of the incident light. The material comprises nanoparticles composed of a nanorod with a nanosphere of 30 nm diameter attached. When diluted into liquids, a small concentration of nanoparticles absorbs on average 26% more than carbon nanotubes, the darkest material available to date. By pumping a dye optical amplifier with nanosecond pulses of 100 mW power, we harness the structural darkness of the material and create a new type of light source, which generates monochromatic emission (5 nm wide) without the need for any resonance. This is achieved through the dynamics of light condensation in which all absorbed electromagnetic energy spontaneously generates single-colour energy pulses. © 2016 Macmillan Publishers Limited. All rights reserved.

  5. Harnessing the Flexibility of Thermostatic Loads in Microgrids with Solar Power Generation

    Directory of Open Access Journals (Sweden)

    Rosa Morales González

    2016-07-01

    Full Text Available This paper presents a demand response (DR framework that intertwines thermodynamic building models with a genetic algorithm (GA-based optimization method. The framework optimizes heating/cooling schedules of end-users inside a business park microgrid with local distributed generation from renewable energy sources (DG-RES based on two separate objectives: net load minimization and electricity cost minimization. DG-RES is treated as a curtailable resource in anticipation of future scenarios where the infeed of DG-RES to the regional distribution network could be limited. We test the DR framework with a case study of a refrigerated warehouse and an office building located in a business park with local PV generation. Results show the technical potential of the DR framework in harnessing the flexibility of the thermal masses from end-user sites in order to: (1 reduce the energy exchange at the point of connection; (2 reduce the cost of electricity for the microgrid end-users; and (3 increase the local utilization of DG-RES in cases where DG-RES exports to the grid are restricted. The results of this work can aid end-users and distribution network operators to reduce energy costs and energy consumption.

  6. Harnessing Innovative Technologies to Advance Children’s Mental Health: Behavioral Parent Training As an Example

    Science.gov (United States)

    Jones, Deborah J.; Forehand, Rex; Cuellar, Jessica; Kincaid, Carlye; Parent, Justin; Fenton, Nicole; Goodrum, Nada

    2012-01-01

    Disruptive behaviors of childhood are among the most common reasons for referral of children to mental health professionals. Behavioral parent training (BPT) is the most efficacious intervention for these problem behaviors, yet BPT is substantially underutilized beyond university research and clinic settings. With the aim of addressing this research-to-practice gap, this article highlights the considerable, but largely unrealized, potential for technology to overcome the two most pressing challenges hindering the diffusion of BPT: (1). The dearth of BPT training and supervision opportunities for therapists who work with families of children with disruptive behaviors and; (2). The failure to engage and retain families in BPT services when services are available. To this end, this review presents a theoretical framework to guide technological innovations in BPT and highlights examples of how technology is currently being harnessed to overcome these challenges. This review also discusses recommendations for using technology as a delivery vehicle to further advance the field of BPT and the potential implications of technological innovations in BPT for other areas of children’s mental health are discussed. PMID:23313761

  7. Harnessing scientific literature reports for pharmacovigilance. Prototype software analytical tool development and usability testing.

    Science.gov (United States)

    Sorbello, Alfred; Ripple, Anna; Tonning, Joseph; Munoz, Monica; Hasan, Rashedul; Ly, Thomas; Francis, Henry; Bodenreider, Olivier

    2017-03-22

    We seek to develop a prototype software analytical tool to augment FDA regulatory reviewers' capacity to harness scientific literature reports in PubMed/MEDLINE for pharmacovigilance and adverse drug event (ADE) safety signal detection. We also aim to gather feedback through usability testing to assess design, performance, and user satisfaction with the tool. A prototype, open source, web-based, software analytical tool generated statistical disproportionality data mining signal scores and dynamic visual analytics for ADE safety signal detection and management. We leveraged Medical Subject Heading (MeSH) indexing terms assigned to published citations in PubMed/MEDLINE to generate candidate drug-adverse event pairs for quantitative data mining. Six FDA regulatory reviewers participated in usability testing by employing the tool as part of their ongoing real-life pharmacovigilance activities to provide subjective feedback on its practical impact, added value, and fitness for use. All usability test participants cited the tool's ease of learning, ease of use, and generation of quantitative ADE safety signals, some of which corresponded to known established adverse drug reactions. Potential concerns included the comparability of the tool's automated literature search relative to a manual 'all fields' PubMed search, missing drugs and adverse event terms, interpretation of signal scores, and integration with existing computer-based analytical tools. Usability testing demonstrated that this novel tool can automate the detection of ADE safety signals from published literature reports. Various mitigation strategies are described to foster improvements in design, productivity, and end user satisfaction.

  8. Harnessing intrinsic localized modes to identify impurities in nonlinear periodic systems

    Science.gov (United States)

    Thota, M.; Harne, R. L.; Wang, K. W.

    2015-02-01

    Intrinsic localized modes (ILMs) are concentrations of vibrational energy in periodic systems/lattices due to the combined influences of nonlinearity and discreteness. Moreover, ILMs can move within the system and may strongly interact with an impurity, such as a stiffness change, mass variation, etc. Numerous scientific fields have uncovered examples and evidence of ILMs, motivating a multidisciplinary pursuit to rigorously understand the underlying principles. In spite of the diverse technical studies, a characterization of ILM interaction behaviors with multiple impurities in dissipative lattices remains outstanding. The insights on such behaviors may be broadly useful when dynamic measurements are the only accessible features of the periodic system. For instance, one may guide an ILM within the lattice using a deliberately applied and steered impurity and harness the observed interaction behaviors with a second, static (immovable) impurity/defect to identify how the underlying lattice is different at the second, defected site, whether or not one knew the position of the defect a priori. In this spirit, this research studies, analyzes, and characterizes the interaction types amongst an ILM and multiple impurities, and devises a method to identify a static defect impurity using quantitatively and qualitatively distinct interaction phenomena. The method is found to be robust to moderate levels of lattice stiffness heterogeneity and is applicable to monitor various property changes that represent impurities. Finally, experimental studies verify that ILMs interact with multiple impurities in unique ways such that defect features may be effectively identified.

  9. A New 3D Printing Strategy by Harnessing Deformation, Instability, and Fracture of Viscoelastic Inks.

    Science.gov (United States)

    Yuk, Hyunwoo; Zhao, Xuanhe

    2018-02-01

    Direct ink writing (DIW) has demonstrated great potential as a multimaterial multifunctional fabrication method in areas as diverse as electronics, structural materials, tissue engineering, and soft robotics. During DIW, viscoelastic inks are extruded out of a 3D printer's nozzle as printed fibers, which are deposited into patterns when the nozzle moves. Hence, the resolution of printed fibers is commonly limited by the nozzle's diameter, and the printed pattern is limited by the motion paths. These limits have severely hampered innovations and applications of DIW 3D printing. Here, a new strategy to exceed the limits of DIW 3D printing by harnessing deformation, instability, and fracture of viscoelastic inks is reported. It is shown that a single nozzle can print fibers with resolution much finer than the nozzle diameter by stretching the extruded ink, and print various thickened or curved patterns with straight nozzle motions by accumulating the ink. A quantitative phase diagram is constructed to rationally select parameters for the new strategy. Further, applications including structures with tunable stiffening, 3D structures with gradient and programmable swelling properties, all printed with a single nozzle are demonstrated. The current work demonstrates that the mechanics of inks plays a critical role in developing 3D printing technology. © 2017 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  10. Harnessing Cell Dynamic Responses on Magnetoelectric Nanocomposite Films to Promote Osteogenic Differentiation.

    Science.gov (United States)

    Tang, Bolin; Zhuang, Junjun; Wang, Liming; Zhang, Bo; Lin, Suya; Jia, Fei; Dong, Lingqing; Wang, Qi; Cheng, Kui; Weng, Wenjian

    2018-03-07

    The binding of cell integrins to proteins adsorbed on the material surface is a highly dynamic process critical for guiding cellular responses. However, temporal dynamic regulation of adsorbed proteins to meet the spatial conformation requirement of integrins for a certain cellular response remains a great challenge. Here, an active CoFe 2 O 4 /poly(vinylidene fluoride-trifluoroethylene) nanocomposite film, which was demonstrated to be an obvious surface potential variation (Δ V ≈ 93 mV) in response to the applied magnetic field intensity (0-3000 Oe), was designed to harness the dynamic binding of integrin-adsorbed proteins by in situ controlling of the conformation of adsorbed proteins. Experimental investigation and molecular dynamics simulation confirmed the surface potential-induced conformational change in the adsorbed proteins. Cells cultured on nanocomposite films indicated that cellular responses in different time periods (adhesion, proliferation, and differentiation) required distinct magnetic field intensity, and synthetically programming the preferred magnetic field intensity of each time period could further enhance the osteogenic differentiation through the FAK/ERK signaling pathway. This work therefore provides a distinct concept that dynamically controllable modulation of the material surface property fitting the binding requirement of different cell time periods would be more conducive to achieving the desired osteogenic differentiation.

  11. Harnessing CRISPR/Cas systems for programmable transcriptional and post-transcriptional regulation

    KAUST Repository

    Mahas, Ahmed

    2017-11-29

    Genome editing has enabled broad advances and novel approaches in studies of gene function and structure; now, emerging methods aim to precisely engineer post-transcriptional processes. Developing precise, efficient molecular tools to alter the transcriptome holds great promise for biotechnology and synthetic biology applications. Different approaches have been employed for targeted degradation of RNA species in eukaryotes, but they lack programmability and versatility, thereby limiting their utility for diverse applications. The CRISPR/Cas9 system has been harnessed for genome editing in many eukaryotic species and, using a catalytically inactive Cas9 variant, the CRISPR/dCas9 system has been repurposed for transcriptional regulation. Recent studies have used other CRISPR/Cas systems for targeted RNA degradation and RNA-based manipulations. For example, Cas13a, a Type VI-A endonuclease, has been identified as an RNA-guided RNA ribonuclease and used for manipulation of RNA. Here, we discuss different modalities for targeted RNA interference with an emphasis on the potential applications of CRISPR/Cas systems as programmable transcriptional regulators for broad uses, including functional biology, biotechnology, and synthetic biology applications.

  12. Harnessing the opportunities of austerity: a detailed mapping of the Greek transportation sector

    Directory of Open Access Journals (Sweden)

    Iraklis Stamos

    2016-12-01

    Full Text Available Since 2009, shortly after the global economic crisis of 2008, Greece has entered into a deep recession phase. The multifaceted presence of austerity is experienced in an increasing number of sectors of the country. The Greek transportation sector is not immune to this state of affairs. The ongoing crisis had a significant impact on its economic (investment, employment, exports-imports turnover as well as its operational (transportation intensity, throughput, performance aspects and capabilities. In this paper, a detailed mapping of these impacts is laid out, correlating transport-related characteristics, trends and estimations with the respective economic ones. Specifically, the paper presents analyses of the Greek passenger and freight transportation, following a data-driven approach over several areas. Findings show a substantial decrease of activities overall. Paradoxically, however, austerity can also be said to offer opportunities, such as the development of innovative, cost-effective and outward-looking business schemes, for handling transportation-related issues. In the face of these opportunities, authorities and stakeholders have recently turned their attention to ways of harnessing them as they may arise. The paper conducts a detailed analysis of these efforts to discover prospects for development in the midst of austerity, and highlights the steps currently being taken in that direction.

  13. LncRNAWiki: harnessing community knowledge in collaborative curation of human long non-coding RNAs

    KAUST Repository

    Ma, L.

    2014-11-15

    Long non-coding RNAs (lncRNAs) perform a diversity of functions in numerous important biological processes and are implicated in many human diseases. In this report we present lncRNAWiki (http://lncrna.big.ac.cn), a wiki-based platform that is open-content and publicly editable and aimed at community-based curation and collection of information on human lncRNAs. Current related databases are dependent primarily on curation by experts, making it laborious to annotate the exponentially accumulated information on lncRNAs, which inevitably requires collective efforts in community-based curation of lncRNAs. Unlike existing databases, lncRNAWiki features comprehensive integration of information on human lncRNAs obtained from multiple different resources and allows not only existing lncRNAs to be edited, updated and curated by different users but also the addition of newly identified lncRNAs by any user. It harnesses community collective knowledge in collecting, editing and annotating human lncRNAs and rewards community-curated efforts by providing explicit authorship based on quantified contributions. LncRNAWiki relies on the underling knowledge of scientific community for collective and collaborative curation of human lncRNAs and thus has the potential to serve as an up-to-date and comprehensive knowledgebase for human lncRNAs.

  14. Harnessing Wisdom of the Crowds Dynamics for Time-dependent Reputation and Ranking

    Science.gov (United States)

    Daly, Elizabeth M.

    The "wisdom of the crowds" is a concept used to describe the utility of harnessing group behaviour, where user opinion evolves over time and the opinion of the masses collectively demonstrates wisdom. Web 2.0 is a new medium where users are not just consumers, but are also contributors. By contributing content to the system, users become part of the network and relationships between users and content can be derived. Example applications are collaborative bookmarking networks such as dcl.icio.us and file sharing applications such as You Tube and Fliekr. These networks rely on user contributed content, described and classified using tags. The wealth of user generated content can be hard to navigate and search due to difficulties in comparing documents with similar tags and the application of traditional information retrieval scoring techniques are limited. Evaluating the time evolving interests of users may be used to derive quality of content. In this chapter, we explore a technique to rank documents based on reputation. The reputation is a combination of the number of bookmarkers, the reputation of the bookmarking user and the time dynamics of the document. Additionally, this reputation measure is extended to take into account the time-dependent, term-dependent reputation of a document. Experimental results and analysis are presented on a large collaborative IBM bookmarking network called Dogear.

  15. Har du ikke en bra feed på Insta, så har du ikke orden på livet ditt!” Unge jenter sin opplevelse av andres selvrepresentasjon i bildedelingstjenester.

    OpenAIRE

    Magnussen, Andrea Bergum

    2017-01-01

    Mastergradsoppgave i digital kommunikasjon og kultur, Avdeling for lærerutdanning og naturvitenskap, Høgskolen i Innlandet, 2017 Norsk: Denne oppgaven undersøker unge jenters opplevelse av selvrepresentasjonen på bildedelingstjenestene Instagram og Snapchat. Funnene er basert på individuelle intervjuer og et fokusgruppeintervju av fem jenter på 14 år. Intervjupersonene har i hovedsak uttrykt seg om deres opplevelse knyttet til kjendiser og influencers sin selvrepresentasjon. Oppgaven ta...

  16. Sykepleiers erfaringer i møte med ungdommer som har anorexia nervosa, på en barne- eller ungdomspsykiatrisk avdeling.

    OpenAIRE

    Vollan, Renate; Andersen, Iselin Margrethe Iserni

    2016-01-01

    Hensikt: Hensikten med oppgaven er å se nærmere på hvilke erfaringer sykepleiere har med å jobbe med ungdommer som har anorexia nervosa. Metode: Vi har anvend åtte kvalitative forskningsartikler for å gjøre et systematisk litteraturstudie. Funnene ble brukt for å belyse problemstillingen. Resultat: Sykepleierne fulgte et gitt behandlingsprogram, som gjorde at de måtte være overvåkende og strenge. Dette gikk på akkord med deres verdier, og at sykepleierne heller ønsket å være omsorgsfull...

  17. Impact of cell type and epitope tagging on heterologous expression of G protein-coupled receptor: a systematic study on angiotensin type II receptor.

    Directory of Open Access Journals (Sweden)

    Lili Jiang

    Full Text Available Despite heterologous expression of epitope-tagged GPCR is widely adopted for functional characterization, there is lacking of systematic analysis of the impact of expression host and epitope tag on GPCR expression. Angiotensin type II (AT2 receptor displays agonist-dependent and -independent activities, coupling to a spectrum of signaling molecules. However, consensus has not been reached on the subcellular distributions, signaling cascades and receptor-mediated actions. To examine the contributions of host cell and epitope tag on receptor expression and activity, epitope-tagged AT2 receptor variants were transiently or stably expressed in HEK293, CHO-K1 and PC12 cells. The epitope-tagged AT2 receptor variants were detected both on the cell membrane and in the perinuclear region. In transiently transfected HEK293 cells, Myc-AT2 existed predominantly as monomer. Additionally, a ladder of ubiquitinated AT2 receptor proteins was detected. By contrast, stably expressed epitope-tagged AT2 receptor variants existed as both monomer and high molecular weight complexes, and the latter was enriched in cell surface. Glycosylation promoted cell surface expression of Myc-AT2 but had no effect on AT2-GFP in HEK293 cells. In cells that stably expressed Myc-AT2, serum starvation induced apoptosis in CHO-K1 cells but not in HEK293 or PC12 cells. Instead, HEK293 and PC12 cells stably expressing Myc-AT2 exhibited partial cell cycle arrest with cells accumulating at G1 and S phases, respectively. Taken together, these results suggest that expression levels, subcellular distributions and ligand-independent constitutive activities of AT2 receptor were cell type-dependent while posttranslational processing of nascent AT2 receptor protein was modulated by epitope tag and mode of expression.

  18. Harnessing the power of Office 365 to provide a social learning environment through a new Student Portal

    OpenAIRE

    Ellison, Adrian; Arora, Mauli

    2013-01-01

    The University of West London (UWL) has harnessed the potential of a Cloud-based suite of collaboration tools (Microsoft Office 365, offered free of charge to higher education institutions) to create a personalised, interactive, social platform to both improve the learning experience and to enable students to combine social learning with their academic study.UWL describes how it engaged with its students to design and refine the requirements for a new student portal, outsourced the developmen...

  19. Impact of ionic and nanoparticle speciation states of silver on light harnessing photosynthetic events in Spirodela polyrhiza.

    Science.gov (United States)

    Shabnam, Nisha; Sharmila, P; Pardha-Saradhi, P

    2017-01-02

    Owing to wide range of applications, nanotechnology is growing expeditiously. Likely negative impact of nanoparticles (NPs), which are inevitably released into our surroundings, on living organisms is of growing concern. Findings presented here are outcome of investigations carried out to evaluate the impact of ionic and NP speciation states of silver on light harnessing photosynthetic events in Spirodela polyrhiza fronds. Fronds exposed to ionic speciation state showed significant decline in PS (photosystem) II efficiency (Fv/Fm; variable fluorescence/maximal fluorescence), while those exposed to silver nanoparticles (Ag-NPs) showed marginal decline. Accordingly, decline in amplitude of Chl a fluorescence transients was sharper in fronds treated with Ag + than those treated with Ag-NPs. Of the various phases Chl a fluorescence transient, J-I phase [which reflects reduction of plastoquinone (PQ) pool] was most sensitive to both Ag + and Ag-NPs. Phenomenological yield models, built using Biolyzer software, revealed that fronds exposed to Ag + possessed significantly lower potential to trap and harness absorbed light energy for photochemical reactions than those exposed to Ag-NPs. Accordingly, dissipation of absorbed light energy as heat was significantly higher in fronds exposed to Ag + than those exposed to Ag-NPs. These findings revealed that NP speciation state of silver is significantly less toxic to light harnessing photosynthetic machinery of S. polyrhiza, compared to ionic speciation state.

  20. The Ismailis’ « War » against Qat in Harâz

    Directory of Open Access Journals (Sweden)

    Peer Gatter

    2007-03-01

    Full Text Available La « guerre » des ismaeiliens contre le qat dans le Haraz.Entre mobilisation communautaire et contraintes politico-économiquesEn 1999, la communauté ismaélienne des Bohra, du massif du .Harâz a lancé une campagne pour remplacer la culture du qat par d’autres activités économiques. Fortement motivée sur le plan religieux, cette campagne visait aussi à la suppression de la consommation du qat. Plus de 200 000 arbustes de qat ont étés arrachés, auxquels on a substitué des caféiers et des manguiers. Les moyens pour générer les revenus alternatifs incluaient aussi l’élevage, l’apiculture, la planification du développement d’artisanat, à quoi s’ajoutaient des projets d’irrigation, des programmes éducatifs et des prêts sans intérêt pour les investissements dans d’autres secteurs économiques. Bien que cette campagne ait été entravée dès le début par une sécheresse sévère ainsi que par l’absence d’aide gouvernementale, il a abouti d’une part à une réduction réelle de la culture et de la consommation du qat, et à la diversification importante de l’activité économique des fermiers. Mais le projet a aussi entrainé une migration accrue de la campagne vers les villes et l’abandon de nombreuses terres cultivées. Cependant, malgré ces limites, ce projet, unique au Yémen par son haut niveau de la mobilisation sociale, apporte des leçons précieuses concernant les contraintes politiques et économiques qui pèsent sur toute tentative de développer une agriculture sans qat dans les montagnes du Yémen.In 1999, the Ismailî   Bohra Community in Yemen’s Harâz mountains launched a drug substitution programme in order to replace the cultivation of qat with other economic and social activities. Motivated by religious concern, this campaign aimed at suppressing the consumption of qat. Over 200,000 qat trees were uprooted and coffee and mango trees planted instead. Alternative means of income

  1. Harnessing Geographic Information Systems (GIS) to enable community-oriented primary care.

    Science.gov (United States)

    Bazemore, Andrew; Phillips, Robert L; Miyoshi, Thomas

    2010-01-01

    settings, particularly if the hurdles of cost and technological expertise are overcome by harnessing new advances in web-based mapping technology.

  2. Molecular breeding of Saccharomyces cerevisiae with high RNA content by harnessing essential ribosomal RNA transcription regulator.

    Science.gov (United States)

    Sasano, Yu; Kariya, Takahiro; Usugi, Shogo; Sugiyama, Minetaka; Harashima, Satoshi

    2017-12-01

    As yeast is commonly used for RNA production, it is industrially important to breed strains with high RNA contents. The upstream activating factor (UAF) plays an important role in transcription of ribosomal RNA (rRNA), a major constituent of intracellular RNA species. Here, we targeted the essential rRNA transcription regulator Rrn5 of Saccharomyces cerevisiae, a component of the UAF complex, and disrupted the genomic RRN5 gene using a helper plasmid carrying an RRN5 gene. Then we isolated nine suppressor mutants (Sup mutants) of RRN5 gene disruption, causing deficiency in rRNA transcription. The Sup mutants had RNA contents of approximately 40% of the wild type level and expansion of rDNA repeats to ca. 400-700 copies. Reintroduction of a functional RRN5 gene into Sup mutants caused a reduction in the number of rDNA repeats to close to the wild type level but did not change RNA content. However, we found that reintroduction of RRN5 into the Sup16 mutant (in which the FOB1 gene encoding the rDNA replication fork barrier site binding protein was disrupted) resulted in a significant increase (17%) in RNA content compared with wild type, although the rDNA repeat copy number was almost identical to the wild type strain. In this case, upregulated transcription of non-transcribed spacers (NTS) occurred, especially in the NTS2 region; this was likely mediated by RNA polymerase II and accounted for the increased RNA content. Thus, we propose a novel breeding strategy for developing high RNA content yeast by harnessing the essential rRNA transcription regulator.

  3. Harnessing Petaflop-Scale Multi-Core Supercomputing for Problems in Space Science

    Science.gov (United States)

    Albright, B. J.; Yin, L.; Bowers, K. J.; Daughton, W.; Bergen, B.; Kwan, T. J.

    2008-12-01

    The particle-in-cell kinetic plasma code VPIC has been migrated successfully to the world's fastest supercomputer, Roadrunner, a hybrid multi-core platform built by IBM for the Los Alamos National Laboratory. How this was achieved will be described and examples of state-of-the-art calculations in space science, in particular, the study of magnetic reconnection, will be presented. With VPIC on Roadrunner, we have performed, for the first time, plasma PIC calculations with over one trillion particles, >100× larger than calculations considered "heroic" by community standards. This allows examination of physics at unprecedented scale and fidelity. Roadrunner is an example of an emerging paradigm in supercomputing: the trend toward multi-core systems with deep hierarchies and where memory bandwidth optimization is vital to achieving high performance. Getting VPIC to perform well on such systems is a formidable challenge: the core algorithm is memory bandwidth limited with low compute-to-data ratio and requires random access to memory in its inner loop. That we were able to get VPIC to perform and scale well, achieving >0.374 Pflop/s and linear weak scaling on real physics problems on up to the full 12240-core Roadrunner machine, bodes well for harnessing these machines for our community's needs in the future. Many of the design considerations encountered commute to other multi-core and accelerated (e.g., via GPU) platforms and we modified VPIC with flexibility in mind. These will be summarized and strategies for how one might adapt a code for such platforms will be shared. Work performed under the auspices of the U.S. DOE by the LANS LLC Los Alamos National Laboratory. Dr. Bowers is a LANL Guest Scientist; he is presently at D. E. Shaw Research LLC, 120 W 45th Street, 39th Floor, New York, NY 10036.

  4. Harnessing landscape heterogeneity for managing future disturbance risks in forest ecosystems.

    Science.gov (United States)

    Seidl, Rupert; Albrich, Katharina; Thom, Dominik; Rammer, Werner

    2018-03-01

    In order to prevent irreversible impacts of climate change on the biosphere it is imperative to phase out the use of fossil fuels. Consequently, the provisioning of renewable resources such as timber and biomass from forests is an ecosystem service of increasing importance. However, risk factors such as changing disturbance regimes are challenging the continuous provisioning of ecosystem services, and are thus a key concern in forest management. We here used simulation modeling to study different risk management strategies in the context of timber production under changing climate and disturbance regimes, focusing on a 8127 ha forest landscape in the Northern Front Range of the Alps in Austria. We show that under a continuation of historical management, disturbances from wind and bark beetles increase by +39.5% on average over 200 years in response to future climate change. Promoting mixed forests and climate-adapted tree species as well as increasing management intensity effectively reduced future disturbance risk. Analyzing the spatial patterns of disturbance on the landscape, we found a highly uneven distribution of risk among stands (Gini coefficients up to 0.466), but also a spatially variable effectiveness of silvicultural risk reduction measures. This spatial variability in the contribution to and control of risk can be used to inform disturbance management: Stands which have a high leverage on overall risk and for which risks can effectively be reduced (24.4% of the stands in our simulations) should be a priority for risk mitigation measures. In contrast, management should embrace natural disturbances for their beneficial effects on biodiversity in areas which neither contribute strongly to landscape-scale risk nor respond positively to risk mitigation measures (16.9% of stands). We here illustrate how spatial heterogeneity in forest landscapes can be harnessed to address both positive and negative effects of changing natural disturbance regimes in

  5. Harnessing long-term flux records to better understand ecosystem response to drought

    Science.gov (United States)

    Novick, K. A.; Ficklin, D. L.; Stoy, P. C.; Williams, C. A.; Bohrer, G.; Oishi, A. C.; Papuga, S. A.; Blanken, P.; Noormets, A.; Scott, R. L.; Wang, L.; Roman, D. T.; Yi, K.; Sulman, B. N.; Phillips, R.

    2016-12-01

    While ongoing climate change affects a number of meteorological drivers relevant to plant functioning, the predicted increase in the frequency and severity of droughts may ultimately have the biggest impact on ecosystem carbon cycling. Because it is difficult to experimentally manipulate all of the meteorological drivers that change during drought (including precipitation, light, temperature, and humidity), our understanding of the mechanisms by which plants respond to drought is generally limited to an understanding of how plants respond to variable soil moisture. As flux tower records grow in length and number, they permit us to harness natural spatial and temporal variability in hydrologic condition to better understand how ecosystems respond to the full suite of meteorological drivers that change during drought stress. Here, a series of case studies are presented that illustrate how long term flux data can be used to disentangle limitations to ecosystem functioning imposed by declining soil moisture as compared to rising atmospheric demand for water during drought. At the site-level, we pair observations from the Morgan-Monroe State Forest Ameriflux tower (active since 1999) with eco-physiological datasets collected during the severe 2012 Midwestern drought. We show that vapor pressure deficit (VPD) limits ecosystem carbon uptake and transpiration as much as soil moisture, but that individual species vary in their sensitivity to these drivers. We then present results from two cross-site Ameriflux syntheses that quantify how VPD as compared to soil moisture limitations to carbon and water cycling vary across broad climate gradients spanning semi-arid to mesic biomes. Informed by these results, we end by highlighting ways that flux network data may be leveraged together with other eco-physiological networks and databases to further expand our understanding of the mechanisms determining ecosystem response to drought.

  6. Harnessing Sun’s Energy with Quantum Dots Based Next Generation Solar Cell

    Directory of Open Access Journals (Sweden)

    Mohammad A. Halim

    2012-12-01

    Full Text Available Our energy consumption relies heavily on the three components of fossil fuels (oil, natural gas and coal and nearly 83% of our current energy is consumed from those sources. The use of fossil fuels, however, has been viewed as a major environmental threat because of their substantial contribution to greenhouse gases which are responsible for increasing the global average temperature. Last four decades, scientists have been searching for alternative sources of energy which need to be environmentally clean, efficient, cost-effective, renewable, and sustainable. One of the promising sustainable sources of energy can be achieved by harnessing sun energy through silicon wafer, organic polymer, inorganic dye, and quantum dots based solar cells. Among them, quantum dots have an exceptional property in that they can excite multiple electrons using only one photon. These dots can easily be synthesized, processed in solution, and incorporated into solar cell application. Interestingly, the quantum dots solar cells can exceed the Shockley-Queisser limit; however, it is a great challenge for other solar cell materials to exceed the limit. Theoretically, the quantum dots solar cell can boost the power conversion efficiency up to 66% and even higher to 80%. Moreover, in changing the size of the quantum dots one can utilize the Sun’s broad spectrum of visible and infrared ranges. This review briefly overviews the present performance of different materials-based solar cells including silicon wafer, dye-sensitized, and organic solar cells. In addition, recent advances of the quantum dots based solar cells which utilize cadmium sulfide/selenide, lead sulfide/selenide, and new carbon dots as light harvesting materials has been reviewed. A future outlook is sketched as to how one could improve the efficiency up to 10% from the current highest efficiency of 6.6%.

  7. The HARNESS Workbench: Unified and Adaptive Access to Diverse HPC Platforms

    Energy Technology Data Exchange (ETDEWEB)

    Sunderam, Vaidy S.

    2012-03-20

    The primary goal of the Harness WorkBench (HWB) project is to investigate innovative software environments that will help enhance the overall productivity of applications science on diverse HPC platforms. Two complementary frameworks were designed: one, a virtualized command toolkit for application building, deployment, and execution, that provides a common view across diverse HPC systems, in particular the DOE leadership computing platforms (Cray, IBM, SGI, and clusters); and two, a unified runtime environment that consolidates access to runtime services via an adaptive framework for execution-time and post processing activities. A prototype of the first was developed based on the concept of a 'system-call virtual machine' (SCVM), to enhance portability of the HPC application deployment process across heterogeneous high-end machines. The SCVM approach to portable builds is based on the insertion of toolkit-interpretable directives into original application build scripts. Modifications resulting from these directives preserve the semantics of the original build instruction flow. The execution of the build script is controlled by our toolkit that intercepts build script commands in a manner transparent to the end-user. We have applied this approach to a scientific production code (Gamess-US) on the Cray-XT5 machine. The second facet, termed Unibus, aims to facilitate provisioning and aggregation of multifaceted resources from resource providers and end-users perspectives. To achieve that, Unibus proposes a Capability Model and mediators (resource drivers) to virtualize access to diverse resources, and soft and successive conditioning to enable automatic and user-transparent resource provisioning. A proof of concept implementation has demonstrated the viability of this approach on high end machines, grid systems and computing clouds.

  8. Harnessing Sun's Energy with Quantum Dots Based Next Generation Solar Cell.

    Science.gov (United States)

    Halim, Mohammad A

    2012-12-27

    Our energy consumption relies heavily on the three components of fossil fuels (oil, natural gas and coal) and nearly 83% of our current energy is consumed from those sources. The use of fossil fuels, however, has been viewed as a major environmental threat because of their substantial contribution to greenhouse gases which are responsible for increasing the global average temperature. Last four decades, scientists have been searching for alternative sources of energy which need to be environmentally clean, efficient, cost-effective, renewable, and sustainable. One of the promising sustainable sources of energy can be achieved by harnessing sun energy through silicon wafer, organic polymer, inorganic dye, and quantum dots based solar cells. Among them, quantum dots have an exceptional property in that they can excite multiple electrons using only one photon. These dots can easily be synthesized, processed in solution, and incorporated into solar cell application. Interestingly, the quantum dots solar cells can exceed the Shockley - Queisser limit; however, it is a great challenge for other solar cell materials to exceed the limit. Theoretically, the quantum dots solar cell can boost the power conversion efficiency up to 66% and even higher to 80%. Moreover, in changing the size of the quantum dots one can utilize the Sun's broad spectrum of visible and infrared ranges. This review briefly overviews the present performance of different materials-based solar cells including silicon wafer, dye-sensitized, and organic solar cells. In addition, recent advances of the quantum dots based solar cells which utilize cadmium sulfide/selenide, lead sulfide/selenide, and new carbon dots as light harvesting materials has been reviewed. A future outlook is sketched as to how one could improve the efficiency up to 10% from the current highest efficiency of 6.6%.

  9. Harnessing Sun’s Energy with Quantum Dots Based Next Generation Solar Cell

    Science.gov (United States)

    Halim, Mohammad A.

    2012-01-01

    Our energy consumption relies heavily on the three components of fossil fuels (oil, natural gas and coal) and nearly 83% of our current energy is consumed from those sources. The use of fossil fuels, however, has been viewed as a major environmental threat because of their substantial contribution to greenhouse gases which are responsible for increasing the global average temperature. Last four decades, scientists have been searching for alternative sources of energy which need to be environmentally clean, efficient, cost-effective, renewable, and sustainable. One of the promising sustainable sources of energy can be achieved by harnessing sun energy through silicon wafer, organic polymer, inorganic dye, and quantum dots based solar cells. Among them, quantum dots have an exceptional property in that they can excite multiple electrons using only one photon. These dots can easily be synthesized, processed in solution, and incorporated into solar cell application. Interestingly, the quantum dots solar cells can exceed the Shockley-Queisser limit; however, it is a great challenge for other solar cell materials to exceed the limit. Theoretically, the quantum dots solar cell can boost the power conversion efficiency up to 66% and even higher to 80%. Moreover, in changing the size of the quantum dots one can utilize the Sun’s broad spectrum of visible and infrared ranges. This review briefly overviews the present performance of different materials-based solar cells including silicon wafer, dye-sensitized, and organic solar cells. In addition, recent advances of the quantum dots based solar cells which utilize cadmium sulfide/selenide, lead sulfide/selenide, and new carbon dots as light harvesting materials has been reviewed. A future outlook is sketched as to how one could improve the efficiency up to 10% from the current highest efficiency of 6.6%. PMID:28348320

  10. Harnessing landscape heterogeneity for managing future disturbance risks in forest ecosystems

    Science.gov (United States)

    Seidl, Rupert; Albrich, Katharina; Thom, Dominik; Rammer, Werner

    2018-01-01

    In order to prevent irreversible impacts of climate change on the biosphere it is imperative to phase out the use of fossil fuels. Consequently, the provisioning of renewable resources such as timber and biomass from forests is an ecosystem service of increasing importance. However, risk factors such as changing disturbance regimes are challenging the continuous provisioning of ecosystem services, and are thus a key concern in forest management. We here used simulation modeling to study different risk management strategies in the context of timber production under changing climate and disturbance regimes, focusing on a 8127 ha forest landscape in the Northern Front Range of the Alps in Austria. We show that under a continuation of historical management, disturbances from wind and bark beetles increase by +39.5% on average over 200 years in response to future climate change. Promoting mixed forests and climate-adapted tree species as well as increasing management intensity effectively reduced future disturbance risk. Analyzing the spatial patterns of disturbance on the landscape, we found a highly uneven distribution of risk among stands (Gini coefficients up to 0.466), but also a spatially variable effectiveness of silvicultural risk reduction measures. This spatial variability in the contribution to and control of risk can be used to inform disturbance management: Stands which have a high leverage on overall risk and for which risks can effectively be reduced (24.4% of the stands in our simulations) should be a priority for risk mitigation measures. In contrast, management should embrace natural disturbances for their beneficial effects on biodiversity in areas which neither contribute strongly to landscape-scale risk nor respond positively to risk mitigation measures (16.9% of stands). We here illustrate how spatial heterogeneity in forest landscapes can be harnessed to address both positive and negative effects of changing natural disturbance regimes in

  11. Epidemiology of musculoskeletal injuries in a population of harness Standardbred racehorses in training

    Science.gov (United States)

    2014-01-01

    Background There is a substantial paucity of studies concerning musculoskeletal injuries in harness Standardbred racehorses. Specifically, little is known about the epidemiology of exercise-related musculoskeletal injuries. Most studies on this subject involve Thoroughbred racehorses, whose biomechanics and racing speed differ from Standardbred, making comparisons difficult. Here, a population of Standardbred racehorses trained at the same racecourse was studied over four years and a classification system for exercise-related musculoskeletal injuries was designed. The incidence rates of musculoskeletal injuries causing horses’ withdrawal from training for 15 days or longer were investigated. A mixed-effects Poisson regression model was used to estimate musculoskeletal injury rates and to describe significance of selected risk factors for exercise-related injuries in this population. Results A total of 356 trotter racehorses from 10 different stables contributed 8961 months at risk of musculoskeletal injuries. Four-hundred-and-twenty-nine injuries were reported and classified into 16 categories, based on their aetiology and anatomical localisation. The overall exercise-related injury rate was 4.79 per 100 horse months. When considering risk factors one by one in separate univariable analyses, we obtained the following results: rates did not differ significantly between genders and classes of age, whereas one driver seemed to cause fewer injuries than the others. Racing speed and racing intensity, as well as recent medical history, seemed to be significant risk factors (p fracture are lower in Standardbreds compared to Thoroughbreds, whereas the opposite is true for tendon and suspensory ligament injuries. In addition to identification of risk factors for musculoskeletal injuries among Standardbred racehorses, results suggest that racing intensity seems to be a protective predictor of risk and recent medical history could be used to identify horses at risk of

  12. Harnessing Science and Technology for preservation and conservation of cultural heritage in Malaysia

    International Nuclear Information System (INIS)

    Adi Taha

    2005-04-01

    Malaysia's heritage is extraordinarily rich. Heritage links people, places and things from our history to the present and to the future. Department of Muzeums and Antiquities work diligently at collecting and preserving the artifacts, written records, oral traditions, special places and lands that make up the Malaysia's history. Over the years our concept of cultural heritage and its role as a central part of the experience of our communities has expanded from a focus on objects and monuments to include our social structures, ways of life, beliefs and systems of knowledge. We seek answers in our attempts to promote the understanding and unity among people that have made our country a nation regardless of ethnic origins and religious affiliations, and to prolong the life essence of our rich heritage. We found a simple but yet, a meaningful answer; Harnessing Science and Technology for Preservation and Conservation of Cultural Heritage in Malaysia. Conservation has gained an increasing importance world over, as there is greater awareness and a sense of urgency about the need to conserve and preserve cultural heritages. Recent years are witnessing unprecedented growth in various fields of science and technology in Malaysia, such as materials technology, medical sciences, biotechnology, information and communications technology. Whichever perspective is used, it is clear that science forms an integral part of Malaysia's culture, in the past as well as now. Fulfilling a vital function as a carrier of knowledge and methodology, sciences places on our shoulders a strong obligation towards future generations. As Malaysians, we have been formed by our cultural heritage. Clearly, we must protect that heritage and continue to enrich and develop it, incorporating new knowledge, new insights, new ideas and new experience. (Author)

  13. Genetic mapping of rust resistance genes in confection sunflower line HA-R6 and oilseed line RHA 397.

    Science.gov (United States)

    Gong, L; Gulya, T J; Markell, S G; Hulke, B S; Qi, L L

    2013-08-01

    Few widely effective resistance sources to sunflower rust, incited by Puccinia helianthi Schwein., have been identified in confection sunflower (Helianthus annuus L.). The USDA inbred line HA-R6 is one of the few confection sunflower lines resistant to rust. A previous allelism test indicated that rust resistance genes in HA-R6 and RHA 397, an oilseed-type restorer line, are either allelic or closely linked; however, neither have been characterized nor molecularly mapped. The objectives of this study are (1) to locate the rust resistance genes in HA-R6 and RHA 397 on a molecular map, (2) to develop closely linked molecular markers for rust resistance diagnostics, and (3) to determine the resistance spectrum of two lines when compared with other rust-resistant lines. Two populations of 140 F2:3 families each from the crosses of HA 89, as susceptible parent, with HA-R6 and RHA 397 were inoculated with race 336 of P. helianthi in the greenhouse. The resistance genes (R-genes) in HA-R6 and RHA 397 were molecularly mapped to the lower end of linkage group 13, which encompasses a large R-gene cluster, and were designated as R 13a and R 13b, respectively. In the initial maps, SSR (simple sequence repeat) and InDel (insertion and deletion) markers revealed 2.8 and 8.2 cM flanking regions for R 13a and R 13b, respectively, linked with a common marker set of four co-segregating markers, ORS191, ORS316, ORS581, and ZVG61, in the distal side and one marker ORS464 in the proximal side. To identify new markers closer to the genes, sunflower RGC (resistance gene candidate) markers linked to the downy mildew R-gene Pl 8 and located at the same region as R 13a and R 13b were selected to screen the two F2 populations. The RGC markers RGC15/16 and a newly developed marker SUN14 designed from a BAC contig anchored by RGC251 further narrowed down the region flanking R 13a and R 13b to 1.1 and 0.1 cM, respectively. Both R 13a and R 13b are highly effective against all rust races

  14. A vaccine grade of yeast Saccharomyces cerevisiae expressing mammalian myostatin

    Directory of Open Access Journals (Sweden)

    Zhang Tingting

    2012-12-01

    Full Text Available Abstract Background Yeast Saccharomyces cerevisiae is a widely-used system for protein expression. We previously showed that heat-killed whole recombinant yeast vaccine expressing mammalian myostatin can modulate myostatin function in mice, resulting in increase of body weight and muscle composition in these animals. Foreign DNA introduced into yeast cells can be lost soon unless cells are continuously cultured in selection media, which usually contain antibiotics. For cost and safety concerns, it is essential to optimize conditions to produce quality food and pharmaceutical products. Results We developed a simple but effective method to engineer a yeast strain stably expressing mammalian myostatin. This method utilized high-copy-number integration of myostatin gene into the ribosomal DNA of Saccharomyces cerevisiae. In the final step, antibiotic selection marker was removed using the Cre-LoxP system to minimize any possible side-effects for animals. The resulting yeast strain can be maintained in rich culture media and stably express mammalian myostatin for two years. Oral administration of the recombinant yeast was able to induce immune response to myostatin and modulated the body weight of mice. Conclusions Establishment of such yeast strain is a step further toward transformation of yeast cells into edible vaccine to improve meat production in farm animals and treat human muscle-wasting diseases in the future.

  15. Harnessing the power of mobile technologies for collaborating, crowdsourcing, and creating

    Science.gov (United States)

    Crompton, H.

    2015-12-01

    Today's digital technologies can have a powerful influence on teaching and learning. M-learning and u-learning in particular are changing pedagogical practice. Sub categories are rapidly emerging, such as context-aware ubiquitous learning, that involve students learning subject content while immersed in authentic and relevant surroundings. Learning cultures are a nebulous blend of traditions, values, beliefs, and rituals built up over time. For a long time, education has long been conceived as classroom-based and predominantly sedentary (Merchant, 2012). Recent mobile technologies are disrupting this culture in favor of learning that is contextualized, personalized, on demand, and ubiquitous (Crompton, 2013). 21st century students are a different breed than past generations (Prensky, 2001). These students have grown up in a time that has not only altered their perceptions and practices but modified the wiring of the brain through neuroplasticity (Crompton, 2012). Students now cognitively receive information quickly through non-linear methods (Gross, 2003, Oblinger & Oblinger, 2005). They think differently. They also seem to be attached to mobile devices 24/7, although the content of the lesson does not match what they seem to be doing on the mobile devices. This presentation will showcase how to get your students to harness the power of mobile devices for educational purposes. For example, students in your classes will be using devices to collaborate on activities with Google Forms, crowdsourcing the best class questions in Slido, and screencasting thoughts and ideas to share with others with Educreations. These are examples of free apps or Web 2.0 tools that can be used on all the major mobile platforms. Crompton, H. (2013). Mobile learning: New approach, new theory. In Z. L. Berge & L. Y. Muilenburg (Eds.), Handbook of mobile learning (pp. 47-57). Prensky, M. (2001). Digital natives, digital immigrants. Mcb University Press, 9(5). Oblinger, D., & Oblinger, J

  16. Harnessing electro driven separation technique for the separation of selected agrochemicals

    International Nuclear Information System (INIS)

    Wan Aini Wan Ibrahim; Alam, S.M. Monjurul; Azli Sulaiman

    2008-01-01

    Electro driven separation techniques offer a different approach to the analysis of complex mixtures than do traditional pressure-driven chromatographic system; it may rely on electrophoresis, the transport of charged species through a medium by an applied field or may rely on electro driven mobile phase to provide a true chromatographic separation. In the current work the potential of an electro driven separation technique viz. Micellar electrokinetic chromatography (MEKC), is harnessed for the separation of selected agrochemicals (organophosphorus pesticides, OPPs) widely used in the agriculture sector in Malaysia. The current study compares the use of MEKC in normal mode (NM) and reverse mode (RM) for the separation of the selected OPPS. This study also highlights the difference in separations produced by performing separations in normal mode-MEKC (NM-MEKC) and reverse mode-MEKC (RM-MEKC) for the selected OPPs. In RM-MEKC, separation is conducted at acidic pH (pH 2.5 in the current work) where the electro osmotic flow (EOF) is weak whereas in NM-MEKC, the separation is carried out under basic pH (9.3 in this work) where the EOF is strong. A reverse migration order of the OPPs was observed under RM-MEKC. Separation under NM-MEKC was found to be superior to those of RM- MEKC. A comparison is also made between separations performed under sweeping-NM-MEKC and sweeping-RM-MEKC. In sweeping, the OPPs are prepared in the same background solution (BGS) minus the micelles and is adjusted to the same conductivity as the BGS. The study showed that NM-MEKC is more sensitive than RM-MEKC but sweeping-RM-MEKC is superior to sweeping-NM-MEKC. However, sweeping-RM-MEKC only separates two of the OPPs in a single run whereas sweeping-NM-MEKC separates four OPPs in a single run. The better choice of separation mode would be sweeping-NM- MEKC for more OPPs separation in a single run. (author)

  17. MicroRNA Expression Profiling to Identify and Validate Reference Genes for the Relative Quantification of microRNA in Rectal Cancer

    DEFF Research Database (Denmark)

    Eriksen, Anne Haahr Mellergaard; Andersen, Rikke Fredslund; Pallisgaard, Niels

    2016-01-01

    INTRODUCTION: MicroRNAs (miRNAs) play important roles in regulating biological processes at the post-transcriptional level. Deregulation of miRNAs has been observed in cancer, and miRNAs are being investigated as potential biomarkers regarding diagnosis, prognosis and prediction in cancer...... management. Real-time quantitative polymerase chain reaction (RT-qPCR) is commonly used, when measuring miRNA expression. Appropriate normalisation of RT-qPCR data is important to ensure reliable results. The aim of the present study was to identify stably expressed miRNAs applicable as normaliser candidates...... in future studies of miRNA expression in rectal cancer.MATERIALS AND METHODS: We performed high-throughput miRNA profiling (OpenArray®) on ten pairs of laser micro-dissected rectal cancer tissue and adjacent stroma. A global mean expression normalisation strategy was applied to identify the most stably...

  18. Developmental dysplasia of the hip in neonates: evolution of acetabular dysplasia after hip stabilization by brief Pavlik harness treatment.

    Science.gov (United States)

    Bin, K; Laville, J-M; Salmeron, F

    2014-06-01

    The recommended treatment duration in neonates with developmental dysplasia of the hip (DDH) varies depending on whether prolonged Pavlik harness therapy is believed to favourably affect the course of the acetabular dysplasia. According to one theory, several months of additional Pavlik harness therapy after achieving hip reduction contributes to correct the acetabular dysplasia. Another theory holds that hip dislocation induces the acetabular dysplasia, which corrects spontaneously once the femoral head is properly seated in the acetabulum. Here, we evaluated this second theory by studying outcomes after early brief Pavlik harness therapy. Acetabular dysplasia associated with neonatal hip instability undergoes self-correction provided stable hip reduction is achieved very early after birth. Therefore, the duration of Pavlik harness therapy can be substantially shortened. We defined hip instability as either reducible hip dislocation or a very easily dislocatable hip with a soft clunk precluding determination of spontaneous hip position as dislocated or reduced. Static and dynamic ultrasound scans were obtained. Patients with ultrasonographic instability (pubo-femoral distance>5mm with less than 50% of coverage) underwent a second physical examination and received treatment. We re-evaluated 42 abnormal hips in 30 patients after a mean follow-up of 6.7 years (range, 5-14 years). Mean age at treatment initiation was 5 days (range, 1-15 days) and mean treatment duration was 34 days (range, 15-75 days). Mean acetabular angle was 20° (range, 12°-30°) and mean Wiberg's lateral centre-edge angle was 30° (range, 22°-35°). Blunting of the lateral angle of the bony roof was noted in 8 hips at last follow-up. In 1 patient whose hip was stable clinically but unstable by ultrasonography at 21 days of age, recurrent dislocation occurred at 5 months of age. The Severin class was 1a in all patients. Despite continuing controversy about whether hip dislocation induces

  19. High- and low-affinity sites for sodium in delta-OR-G(i)1 alpha (Cys(351)-Ile(351)) fusion protein stably expressed in HEK293 cells; functional significance and correlation with biophysical state of plasma membrane

    Czech Academy of Sciences Publication Activity Database

    Vošahlíková, Miroslava; Jurkiewicz, Piotr; Roubalová, Lenka; Hof, Martin; Svoboda, Petr

    2014-01-01

    Roč. 387, č. 5 (2014), s. 487-502 ISSN 0028-1298 R&D Projects: GA ČR(CZ) GAP207/12/0919; GA ČR(CZ) GBP304/12/G069 Institutional support: RVO:67985823 ; RVO:61388955 Keywords : delta - opioid receptor * monovalent ions * agonist and antagonist binding * [35S]GTPγS binding * membrane biophysics * Laurdan fluorescence Subject RIV: CE - Biochemistry Impact factor: 2.471, year: 2014

  20. The influence of monovalent cations on trimeric G protein Gi1alfa activity in HEK293 cells stably expressing DOR-Gi1alfa (Cys351-Ile351) fusion protein

    Czech Academy of Sciences Publication Activity Database

    Vošahlíková, Miroslava; Svoboda, Petr

    2011-01-01

    Roč. 60, č. 3 (2011), s. 541-547 ISSN 0862-8408 R&D Projects: GA AV ČR(CZ) IAA500110606; GA MŠk(CZ) LC554; GA ČR(CZ) GD305/08/H037 Institutional research plan: CEZ:AV0Z50110509 Keywords : delta-opioid receptor (DOR) * monovalent ions * G(i)1alfa protein Subject RIV: CE - Biochemistry Impact factor: 1.555, year: 2011

  1. A protein phosphatase methylesterase (PME-1) is one of several novel proteins stably associating with two inactive mutants of protein phosphatase 2A.

    Science.gov (United States)

    Ogris, E; Du, X; Nelson, K C; Mak, E K; Yu, X X; Lane, W S; Pallas, D C

    1999-05-14

    Carboxymethylation of proteins is a highly conserved means of regulation in eukaryotic cells. The protein phosphatase 2A (PP2A) catalytic (C) subunit is reversibly methylated at its carboxyl terminus by specific methyltransferase and methylesterase enzymes which have been purified, but not cloned. Carboxymethylation affects PP2A activity and varies during the cell cycle. Here, we report that substitution of glutamine for either of two putative active site histidines in the PP2A C subunit results in inactivation of PP2A and formation of stable complexes between PP2A and several cellular proteins. One of these cellular proteins, herein named protein phosphatase methylesterase-1 (PME-1), was purified and microsequenced, and its cDNA was cloned. PME-1 is conserved from yeast to human and contains a motif found in lipases having a catalytic triad-activated serine as their active site nucleophile. Bacterially expressed PME-1 demethylated PP2A C subunit in vitro, and okadaic acid, a known inhibitor of the PP2A methylesterase, inhibited this reaction. To our knowledge, PME-1 represents the first mammalian protein methylesterase to be cloned. Several lines of evidence indicate that, although there appears to be a role for C subunit carboxyl-terminal amino acids in PME-1 binding, amino acids other than those at the extreme carboxyl terminus of the C subunit also play an important role in PME-1 binding to a catalytically inactive mutant.

  2. Immunosenescent CD57+CD4+ T-cells Accumulate and Contribute to Interferon-γ Responses in HIV Patients Responding Stably to ART

    Directory of Open Access Journals (Sweden)

    Sonia Fernandez

    2011-01-01

    Full Text Available HIV-infected individuals responding to antiretroviral therapy (ART after severe CD4+ T-cell depletion may retain low responses to recall antigens [eg: cytomegalovirus (CMV] and altered expression of T-cell co-stimulatory molecules consistent with immunosenescence. We investigated the capacity of phenotypically senescent cells to generate cytokines in HIV patients receiving long-term ART (n = 18 and in healthy controls (n = 10. Memory T-cells were assessed by interferon (IFN-γ ELISpot assay and flow cytometrically via IFN-γ or IL-2. Proportions of CD57brightCD28null CD4+ T-cells correlated with IFN-γ responses to CMV (p = 0.009 and anti-CD3 (p = 0.002 in HIV patients only. Proportions of CD57brightCD28null CD8+ T-cells and CD8+ T-cell IFN-γ responses to CMV peptides correlated in controls but not HIV patients. IL-2 was predominantly produced by CD28+T-cells from all donors, whereas IFN-γ was mostly produced by CD57+ T-cells. The findings provide evidence of an accumulation of immunosenescent T-cells able to make IFN-γ. This may influence the pathogenesis of secondary viral infections in HIV patients receiving ART.

  3. Determining Damping Trends from a Range of Cable Harness Assemblies on a Launch Vehicle Panel from Test Measurements

    Science.gov (United States)

    Smith, Andrew; Davis, R. Ben; LaVerde, Bruce; Jones, Douglas

    2012-01-01

    The team of authors at Marshall Space Flight Center (MSFC) has been investigating estimating techniques for the vibration response of launch vehicle panels excited by acoustics and/or aero-fluctuating pressures. Validation of the approaches used to estimate these environments based on ground tests of flight like hardware is of major importance to new vehicle programs. The team at MSFC has recently expanded upon the first series of ground test cases completed in December 2010. The follow on tests recently completed are intended to illustrate differences in damping that might be expected when cable harnesses are added to the configurations under test. This validation study examines the effect on vibroacoustic response resulting from the installation of cable bundles on a curved orthogrid panel. Of interest is the level of damping provided by the installation of the cable bundles and whether this damping could be potentially leveraged in launch vehicle design. The results of this test are compared with baseline acoustic response tests without cables. Damping estimates from the measured response data are made using a new software tool that employs a finite element model (FEM) of the panel in conjunction with advanced optimization techniques. This paper will report on the \\damping trend differences. observed from response measurements for several different configurations of cable harnesses. The data should assist vibroacoustics engineers to make more informed damping assumptions when calculating vibration response estimates when using model based analysis approach. Achieving conservative estimates that have more flight like accuracy is desired. The paper may also assist analysts in determining how ground test data may relate to expected flight response levels. Empirical response estimates may also need to be adjusted if the measured response used as an input to the study came from a test article without flight like cable harnesses.

  4. The Elusive Path of Brain Tissue Oxygenation and Cerebral Perfusion in Harness Hang Syncope in Mountain Climbers.

    Science.gov (United States)

    Lanfranconi, Francesca; Pollastri, Luca; Corna, Giovanni; Bartesaghi, Manuela; Novarina, Massimiliano; Ferri, Alessandra; Miserocchi, Giuseppe Andrea

    2017-12-01

    Lanfranconi, Francesca, Luca Pollastri, Giovanni Corna, Manuela Bartesaghi, Massimiliano Novarina, Alessandra Ferri, and Giuseppe Andrea Miserocchi. The elusive path of brain tissue oxygenation and cerebral perfusion in harness hang syncope in mountain climbers. High Alt Med Biol. 18:363-371, 2017. Harness hang syncope (HHS) is a risk that specifically affects wide ranges of situations requiring safety harnesses in mountains. An irreversible orthostatic stasis could lead to death if a prompt rescue is not performed. We aimed at evaluating the risk of developing HHS and at identifying the characteristics related to the pathogenesis of HHS. Forty adults (aged 39.1 [8.2] years) were enrolled in a suspension test lasting about 28.7 (11.4) minutes. We measured cardiovascular parameters, and near infrared spectroscopy (NIRS) was used to assess cerebral hypoxia by changes in the concentration of oxyhemoglobin (Δ[HbO 2 ]) and de-oxyhemoglobin (Δ[HHb]). In the four participants who developed HHS: (1) systolic and diastolic blood pressure showed ample oscillations with a final abrupt drop (∼30 mmHg); (2) Δ[HbO 2 ] increased after 8-12 minutes of suspension and reached a plateau before HHS; and (3) Δ[HHb] decreased with a final abrupt increase before syncope. Participants who developed HHS failed to activate cardiovascular reflexes that usually safeguard O 2 availability to match the metabolic needs of the brain tissue. Since cerebral hypoxia was detected as an early phenomenon by Δ[HbO 2 ] and Δ[HHb] changes, NIRS measurement appears to be the most important parameter to monitor the onset of HHS.

  5. Harnessing the Risk-Related Data Supply Chain: An Information Architecture Approach to Enriching Human System Research and Operations Knowledge

    Science.gov (United States)

    Buquo, Lynn E.; Johnson-Throop, Kathy A.

    2011-01-01

    An Information Architecture facilitates the understanding and, hence, harnessing of the human system risk-related data supply chain which enhances the ability to securely collect, integrate, and share data assets that improve human system research and operations. By mapping the risk-related data flow from raw data to useable information and knowledge (think of it as a data supply chain), the Human Research Program (HRP) and Space Life Science Directorate (SLSD) are building an information architecture plan to leverage their existing, and often shared, IT infrastructure.

  6. Fra skadet barn til kriminell voksen. Hvorvidt har oppvekstsvilkår betydning for senere kriminell adferd?

    OpenAIRE

    2017-01-01

    Oppgaven baserer seg hovedsakelig på hypoteser om hvordan omsorgssvikt og uhensiktsmessige oppvekstsvilkår, kan føre til utvikling av psykiske vansker og kriminelle adferdsmønstre. Fra skadet barn til kriminell voksen, tar utgangspunkt i tidligere undersøkelser gjort blant domfelte i norske fengsler, som kan vise til at 73 prosent har tegn til personlighetsforstyrrelser og at mishandling er en risikofaktor som øker sjansen med å begå en forbrytelse med 50 prosent. I tillegg til en signifikant...

  7. Validation of housekeeping genes for studying differential gene expression in the bovine myometrium.

    Science.gov (United States)

    Rekawiecki, Robert; Kowalik, Magdalena K; Kotwica, Jan

    2013-12-01

    The aim of this study was to determine the steady-state expression of 13 selected housekeeping genes in the myometrium of cyclic and pregnant cows. Cells taken from bovine myometrium on days 1-5, 6-10, 11-16 and 17-20 of the oestrous cycle and in weeks 3-5, 6-8 and 9-12 of pregnancy were used. Reverse transcribed RNA was amplified in real-time PCR using designed primers. Reaction efficiency was determined with the Linreg programme. The geNorm and NormFinder programmes were used to select the best housekeeping genes. They calculate the expression stability factor for each used housekeeping gene with the smallest value for most stably expressed genes. According to geNorm, the most stable housekeeping genes in the myometrium were C2orf29, TPB and TUBB2B, while the least stably expressed genes were 18S RNA, HPRT1 and GAPDH. NormFinder identified the best genes in the myometrium as C2orf29, MRPL12 and TBP, while the worst genes were 18S RNA, B2M and SF3A1. Differences in stability factors between the two programmes may also indicate that the physiological status of the female, e.g. pregnancy, affects the stability of expression of housekeeping genes. The different expression stability of housekeeping genes did not affect progesterone receptor expression but it could be important if small differences in gene expression were measured between studies.

  8. Three decades of harnessing the GPS data explosion for geophysics (Vening Meinesz Medal Lecture)

    Science.gov (United States)

    Blewitt, Geoffrey

    2015-04-01

    In this presentation, I attempt to convey the immensity of the task that faced the geodesy community three decades ago, and continues to challenge us, to harness all potentially valuable GPS data available in the world for geophysical science. It would be fair to see that three decades ago, we were struggling with controlled tests just to get GPS geodesy working, and had little time to imagine the flood of data today. Yet the geodesy community has succeeded in meeting this challenge. Today, for example, the Nevada Geodetic Laboratory produces and makes publicly available coordinate time series for over 12,000 geodetic GPS station around the globe with various data intervals, latencies, and reference frames. About 8,000 stations have their daily time series updated every week, with 4,000 being updated the next day with coordinates at daily and 5 minute intervals. About 2,000 stations have their time series updated every hour with coordinates at 5 minute intervals. I will show examples of how these time series are being used by NGL and many other scientists to study a wide variety of geophysical topics, including plate tectonics, earthquake modeling, seismic and tsunami hazard, volcanic deformation, water resources, mountain growth, terrestrial reference frame realization, glacial isostatic adjustment, ice sheet melting, sea level rise and coastal subsidence, and even fundamental physics, using GPS atomic clocks to probe the nature of dark matter in the universe. The explosion in GPS data has challenged us to invent new data processing algorithms and develop robust automation in order to keep up with the flood. This explosion has been exponential, and therefore it can be said that it is not a recent phenomena, but rather that it began in the earliest years of GPS geodesy, and has always posed a challenge to us. Over the course of my post-doctoral career starting in late 1985, I have had the good fortune to witness the key developments that have taken place since the

  9. Harness That S.O.B.: Distributing Remote Sensing Analysis in a Small Office/Business

    Science.gov (United States)

    Kramer, J.; Combe, J.; McCord, T. B.

    2009-12-01

    Researchers in a small office/business (SOB) operate with limited funding, equipment, and software availability. To mitigate these issues, we developed a distributed computing framework that: 1) leverages open source software to implement functionality otherwise reliant on proprietary software and 2) harnesses the unused power of (semi-)idle office computers with mixed operating systems (OSes). This abstract outlines some reasons for the effort, its conceptual basis and implementation, and provides brief speedup results. The Multiple-Endmember Linear Spectral Unmixing Model (MELSUM)1 processes remote-sensing (hyper-)spectral images. The algorithm is computationally expensive, sometimes taking a full week or more for a 1 million pixel/100 wavelength image. Analysis of pixels is independent, so a large benefit can be gained from parallel processing techniques. Job concurrency is limited by the number of active processing units. MELSUM was originally written in the Interactive Data Language (IDL). Despite its multi-threading capabilities, an IDL instance executes on a single machine, and so concurrency is limited by the machine's number of central processing units (CPUs). Network distribution can access more CPUs to provide a greater speedup, while also taking advantage of (often) underutilized extant equipment. appropriately integrating open source software magnifies the impact by avoiding the purchase of additional licenses. Our method of distribution breaks into four conceptual parts: 1) the top- or task-level user interface; 2) a mid-level program that manages hosts and jobs, called the distribution server; 3) a low-level executable for individual pixel calculations; and 4) a control program to synchronize sequential sub-tasks. Each part is a separate OS process, passing information via shell commands and/or temporary files. While the control and low-level executables are short-lived, the top-level program and distribution server run (at least) for the entirety of

  10. Housekeeping genes for quantitative expression studies in the three-spined stickleback Gasterosteus aculeatus

    Directory of Open Access Journals (Sweden)

    Becker Sven

    2008-01-01

    Full Text Available Abstract Background During the last years the quantification of immune response under immunological challenges, e.g. parasitation, has been a major focus of research. In this context, the expression of immune response genes in teleost fish has been surveyed for scientific and commercial purposes. Despite the fact that it was shown in teleostei and other taxa that the gene for beta-actin is not the most stably expressed housekeeping gene (HKG, depending on the tissue and experimental treatment, the gene has been used as a reference gene in such studies. In the three-spined stickleback, Gasterosteus aculeatus, other HKG than the one for beta-actin have not been established so far. Results To establish a reliable method for the measurement of immune gene expression in Gasterosteus aculeatus, sequences from the now available genome database and an EST library of the same species were used to select oligonucleotide primers for HKG, in order to perform quantitative reverse-transcription (RT PCR. The expression stability of ten candidate reference genes was evaluated in three different tissues, and in five parasite treatment groups, using the three algorithms BestKeeper, geNorm and NormFinder. Our results showed that in most of the tissues and treatments HKG that could not be used so far due to unknown sequences, proved to be more stably expressed than the one for beta-actin. Conclusion As they were the most stably expressed genes in all tissues examined, we suggest using the genes for the L13a ribosomal binding protein and ubiquitin as alternative or additional reference genes in expression analysis in Gasterosteus aculeatus.

  11. Housekeeping genes for quantitative expression studies in the three-spined stickleback Gasterosteus aculeatus.

    Science.gov (United States)

    Hibbeler, Sascha; Scharsack, Joern P; Becker, Sven

    2008-01-29

    During the last years the quantification of immune response under immunological challenges, e.g. parasitation, has been a major focus of research. In this context, the expression of immune response genes in teleost fish has been surveyed for scientific and commercial purposes. Despite the fact that it was shown in teleostei and other taxa that the gene for beta-actin is not the most stably expressed housekeeping gene (HKG), depending on the tissue and experimental treatment, the gene has been used as a reference gene in such studies. In the three-spined stickleback, Gasterosteus aculeatus, other HKG than the one for beta-actin have not been established so far. To establish a reliable method for the measurement of immune gene expression in Gasterosteus aculeatus, sequences from the now available genome database and an EST library of the same species were used to select oligonucleotide primers for HKG, in order to perform quantitative reverse-transcription (RT) PCR. The expression stability of ten candidate reference genes was evaluated in three different tissues, and in five parasite treatment groups, using the three algorithms BestKeeper, geNorm and NormFinder. Our results showed that in most of the tissues and treatments HKG that could not be used so far due to unknown sequences, proved to be more stably expressed than the one for beta-actin. As they were the most stably expressed genes in all tissues examined, we suggest using the genes for the L13a ribosomal binding protein and ubiquitin as alternative or additional reference genes in expression analysis in Gasterosteus aculeatus.

  12. The assessment of global thermo-energy performances of existing district heating systems optimized by harnessing renewable energy sources

    Science.gov (United States)

    Şoimoşan, Teodora M.; Danku, Gelu; Felseghi, Raluca A.

    2017-12-01

    Within the thermo-energy optimization process of an existing heating system, the increase of the system's energy efficiency and speeding-up the transition to green energy use are pursued. The concept of multi-energy district heating system, with high harnessing levels of the renewable energy sources (RES) in order to produce heat, is expected to be the key-element in the future urban energy infrastructure, due to the important role it can have in the strategies of optimizing and decarbonizing the existing district heating systems. The issues that arise are related to the efficient integration of different technologies of harnessing renewable energy sources in the energy mix and to the increase of the participation levels of RES, respectively. For the holistic modeling of the district heating system, the concept of the energy hub was used, where the synergy of different primary forms of entered energy provides the system a high degree energy security and flexibility in operation. The optimization of energy flows within the energy hub allows the optimization of the thermo-energy district system in order to approach the dual concept of smart city & smart energy.

  13. Construction of a rhizosphere pseudomonad with potential to degrade polychlorinated biphenyls and detection of bph gene expression in the rhizosphere.

    OpenAIRE

    Brazil, G M; Kenefick, L; Callanan, M; Haro, A; de Lorenzo, V; Dowling, D N; O'Gara, F

    1995-01-01

    The genetically engineered transposon TnPCB, contains genes (bph) encoding the biphenyl degradative pathway. TnPCB was stably inserted into the chromosome of two different rhizosphere pseudomonads. One genetically modified strain, Pseudomonas fluorescens F113pcb, was characterized in detail and found to be unaltered in important parameters such as growth rate and production of secondary metabolites. The expression of the heterologous bph genes in F113pcb was confirmed by the ability of the ge...

  14. Harnessing Novel Secreted Inhibitors of EGF Receptor Signaling for Breast Cancer Treatment

    Science.gov (United States)

    2008-04-01

    Clemmons, D. R. (2001) Endocr. Rev. 22, 800–817 47. Siwanowicz, I., Popowicz, G. M., Wisniewska, M., Huber , R., Kuenkele, K. P., Lang, K., Engh, R. A...Cell 10, 39-50 (2006). 25. Fujimoto , N. et al. High expression of ErbB family members and their ligands in lung adenocarcinomas that are sensitive to

  15. Engineering low-temperature expression systems for heterologous production of cold-adapted enzymes.

    Science.gov (United States)

    Bjerga, Gro Elin Kjæreng; Lale, Rahmi; Williamson, Adele Kim

    2016-01-01

    Production of psychrophilic enzymes in the commonly used mesophilic expression systems is hampered by low intrinsic stability of the recombinant enzymes at the optimal host growth temperatures. Unless strategies for low-temperature expression are advanced, research on psychrophilic enzymes may end up being biased toward those that can be stably produced in commonly used mesophilic host systems. Two main strategies are currently being explored for the development of low-temperature expression in bacterial hosts: (i) low-temperature adaption of existing mesophilic expression systems, and (ii) development of new psychrophilic hosts. These developments include genetic engineering of the expression cassettes to optimize the promoter/operator systems that regulate heterologous expression. In this addendum we present our efforts in the development of such low-temperature expression systems, and speculate about future advancements in the field and potential applications.

  16. Pre-Exposure Gene Expression in Baboons with and without Pancytopenia after Radiation Exposure

    Directory of Open Access Journals (Sweden)

    Matthias Port

    2017-03-01

    Full Text Available Radiosensitivity differs in humans and likely among primates. The reasons are not well known. We examined pre-exposure gene expression in baboons (n = 17 who developed haematologic acute radiation syndrome (HARS without pancytopenia or a more aggravated HARS with pancytopenia after irradiation. We evaluated gene expression in a two stage study design where stage I comprised a whole genome screen for messenger RNAs (mRNA (microarray and detection of 667 microRNAs (miRNA (real-time quantitative polymerase chain reaction (qRT-PCR platform. Twenty candidate mRNAs and nine miRNAs were selected for validation in stage II (qRT-PCR. None of the mRNA species could be confirmed during the validation step, but six of the nine selected candidate miRNA remained significantly different during validation. In particular, miR-425-5p (receiver operating characteristic = 0.98; p = 0.0003 showed nearly complete discrimination between HARS groups with and without pancytopenia. Target gene searches of miR-425-5p identified new potential mRNAs and associated biological processes linked with radiosensitivity. We found that one miRNA species examined in pre-exposure blood samples was associated with HARS characterized by pancytopenia and identified new target mRNAs that might reflect differences in radiosensitivity of irradiated normal tissue.

  17. Den koreanske bølgen - En kulturvitenskapelig studie av sørkoreansk populærkultur, og hvordan den har nådd Vesten

    OpenAIRE

    Kvinge, Kristin Øyvindsdatter

    2016-01-01

    Hensikten med denne oppgaven er å se på den koreanske bølgen, som ofte refereres til som Hallyu. Den tar for seg både Hallyu som innhold og Hallyu som fenomen. Den koreanske bølgen har spredd seg hurtig i løpet av de siste 20 årene, noe som har bidratt til å gjøre Korea til en av de hurtigst økende økonomiene i den moderne tid. Denne oppgaven forsøker å se nærmere på hvordan Hallyu har nådd Vesten, med fokus på Europa og Norge. Før analysen vil oppgaven se nærmere på hv...

  18. Validation of endogenous normalizing genes for expression analyses in adult human testis and germ cell neoplasms

    DEFF Research Database (Denmark)

    Svingen, T; Jørgensen, Anne; Rajpert-De Meyts, E

    2014-01-01

    The measurement of gene expression levels in cells and tissues typically depends on a suitable point of reference for inferring biological relevance. For quantitative (or real-time) RT-PCR assays, the method of choice is often to normalize gene expression data to an endogenous gene that is stably...... to define suitable normalizing genes for specific cells and tissues. Here, we report on the performance of a panel of nine commonly employed normalizing genes in adult human testis and testicular pathologies. Our analyses revealed significant variability in transcript abundance for commonly used normalizers...

  19. The effect of a 3-point harness restraint and car seat in whiplash-type lateral impacts.

    Science.gov (United States)

    Kumar, Shrawan; Ferrari, Robert; Narayan, Yogesh; Jones, Troy

    2006-01-01

    Seventeen healthy volunteers were subjected to right and left lateral impacts 5.0, 6.8, 9.2, and 16.8 m/s acceleration while positioned in a Volvo car seat with lap and shoulder seat belt restraint in laboratory setting. The purpose of this study was to determine the effect of using a standard 3-point lap and shoulder seat belt and Volvo car seat on the response of the cervical muscles to increasing low-velocity lateral impacts. A previous study of lateral impacts in a 5-point harness restraint with head and trunk in neutral posture suggests that the burden of impact is borne primarily by the splenius capitis muscle contralateral to the direction of impact. That study, however, used a nonstandard harness for automobiles, and other studies suggest that a lap-and-shoulder seat belt may increase the risk of whiplash injury. Triaxial accelerometers recorded the acceleration of the 1) sled, 2) torso at the shoulder level, and 3) head of the participant, while bilateral electromyograms of the sternocleidomastoids, trapezii, and splenii capitis were also recorded. For participants experiencing a right or left lateral impact, the muscle responses increased with increasing levels of acceleration (P trend to progressively decrease with increasing levels of acceleration. The peak head accelerations relative to the sled ranged from 2.5 to 10.6 m/s. When the impact was a right lateral impact, at the highest sled acceleration, the left splenius capitis generated 47% of its maximal voluntary contraction (MVC), and the left trapezius also 46% of its MVC; the left and right sternocleidomastoid, right splenius capitis, and right trapezius generated 29% or less of their MVC. For the highest level of acceleration in a left lateral impact, the right splenius capitis generated 48% of its MVC and the right trapezius 57% of the MVC, the left and right sternocleidomastoid, left splenius capitis, and left trapezius generated 29% or less of their MVC. In both directions of impact, the

  20. Införandet av IAS 41 : Hur har kapitalfördelningen mellan intressenter i den norska laxodlingsbranschen utvecklats?

    OpenAIRE

    Dahlén, Jonas; Josefsson, Carl

    2013-01-01

    Införandet av IAS 41 medförde att redovisningsprinciperna för värderingen av ett företags biologiska tillgångar förändrades. Det finns forskning på hur införandet av IAS 41 påverkat olika aktörer inom skogsindustrin men lite är skrivet om dess påverkan på laxodlingsindustrin. Denna uppsats ämnar undersöka hur kapitalfördelningen till intressenter utvecklats sedan införandet av IAS 41. Studien fokuserar på intressenterna ägare och anställda. Vi har tillämpat en kvantitativ metod där vår empiri...

  1. Harnessing cognitive neuroscience to develop new treatments for improving cognition in schizophrenia: CNTRICS selected cognitive paradigms for animal models.

    Science.gov (United States)

    Moore, Holly; Geyer, Mark A; Carter, Cameron S; Barch, Deanna M

    2013-11-01

    Over the past two decades, the awareness of the disabling and treatment-refractory effects of impaired cognition in schizophrenia has increased dramatically. In response to this still unmet need in the treatment of schizophrenia, the Cognitive Neuroscience Treatment Research to Improve Cognition in Schizophrenia (CNTRICS) initiative was developed. The goal of CNTRICS is to harness cognitive neuroscience to develop a brain-based set of tools for measuring cognition in schizophrenia and to test new treatments. CNTRICS meetings focused on development of tasks with cognitive construct validity for use in both human and animal model studies. This special issue presents papers discussing the cognitive testing paradigms selected by CNTRICS for animal model systems. These paradigms are designed to measure cognitive constructs within the domains of perception, attention, executive function, working memory, object/relational long-term memory, and social/affective processes. Copyright © 2013. Published by Elsevier Ltd.

  2. Study of Green Shipping Technologies - Harnessing Wind, Waves and Solar Power in New Generation Marine Propulsion Systems

    Directory of Open Access Journals (Sweden)

    Grzegorz Rutkowski

    2016-12-01

    Full Text Available The purpose and scope of this paper is to describe the complexity of the new generation marine propulsion technologies implemented in the shipping industry to promote green ships concept and change the view of sea transportation to a more ecological and environment-friendly. Harnessing wind, waves and solar power in shipping industry can help the ship’s owners reduce the operational costs. Reducing fuel consumption results in producing less emissions and provides a clean source of renewable energy. Green shipping technologies can also effectively increase the operating range of vessels and help drive sea transportation towards a greener future and contribute to the global reduction of harmful gas emissions from the world's shipping fleets.

  3. Harnessing the hydropower potential in Africa: What should be the place and role of Grand Inga hydropower project?

    Energy Technology Data Exchange (ETDEWEB)

    Fall, Latsoucabe

    2010-09-15

    Harnessing Africa's huge hydropower potential should be made a priority for the sustainable development of the Continent. Particularly, Grand Inga hydropower project in DRC, due to its gigantic size (40,000 MW) and favourable natural characteristics, could be 'Africa's flagship Project of the 21st Century' offering enormous comparative advantages and opportunities for the benefits of the entire African Continent. Nevertheless, to make it a feasible and palpable 'Model Project', capable of producing clean and affordable energy, the paper tries to respond to key questions on the several daunting challenges to address for its sustainable, cost-effective and timely development and operation.

  4. Turbulent structure of stably stratified inhomogeneous flow

    Science.gov (United States)

    Iida, Oaki

    2018-04-01

    Effects of buoyancy force stabilizing disturbances are investigated on the inhomogeneous flow where disturbances are dispersed from the turbulent to non-turbulent field in the direction perpendicular to the gravity force. Attaching the fringe region, where disturbances are excited by the artificial body force, a Fourier spectral method is used for the inhomogeneous flow stirred at one side of the cuboid computational box. As a result, it is found that the turbulent kinetic energy is dispersed as layered structures elongated in the streamwise direction through the vibrating motion. A close look at the layered structures shows that they are flanked by colder fluids at the top and hotter fluids at the bottom, and hence vertically compressed and horizontally expanded by the buoyancy related to the countergradient heat flux, though they are punctuated by the vertical expansion of fluids at the forefront of the layered structures, which is related to the downgradient heat flux, indicating that the layered structures are gravity currents. However, the phase between temperature fluctuations and vertical velocity is shifted by π/2 rad, indicating that temperature fluctuations are generated by the propagation of internal gravity waves.

  5. Turbulent Mixing in Stably Stratified Flows

    Science.gov (United States)

    2008-03-01

    Turbulent fluid motions are typically characterized by several features including randomness in both space and time, vorticity, an energy cascade ...drawback of this method is that the portion of the flow identified as a turbulent structure is dependent on the type of wavelet filter used (e.g., Haar ...the mesoscale variability of the atmosphere. J. Atmos. Sci., 40:749-761, 1983. E. Lindborg. The energy cascade in a strongly stratified fluid. J

  6. Harnessing collective knowledge to create global public goods for education and health.

    Science.gov (United States)

    Bernardo, Theresa M

    2007-01-01

    Our global interconnectedness has first been expressed in the transformation of business and entertainment, but it will have profound effects on all aspects of our lives, including education and health. Today's students are accustomed to supplying and consuming user-generated content, such as the videos of YouTube, and to using social networking applications, such as MySpace. This can be turned to their advantage in the development of work skills for the twenty-first century. Central among these skills will be the ability to work in teams that cross disciplinary, cultural, and geographic borders.

  7. Microbes at Surface-Air Interfaces: The Metabolic Harnessing of Relative Humidity, Surface Hygroscopicity, and Oligotrophy for Resilience

    Science.gov (United States)

    Stone, Wendy; Kroukamp, Otini; Korber, Darren R.; McKelvie, Jennifer; Wolfaardt, Gideon M.

    2016-01-01

    The human environment is predominantly not aqueous, and microbes are ubiquitous at the surface-air interfaces with which we interact. Yet microbial studies at surface-air interfaces are largely survival-oriented, whilst microbial metabolism has overwhelmingly been investigated from the perspective of liquid saturation. This study explored microbial survival and metabolism under desiccation, particularly the influence of relative humidity (RH), surface hygroscopicity, and nutrient availability on the interchange between these two phenomena. The combination of a hygroscopic matrix (i.e., clay or 4,000 MW polyethylene glycol) and high RH resulted in persistent measurable microbial metabolism during desiccation. In contrast, no microbial metabolism was detected at (a) hygroscopic interfaces at low RH, and (b) less hygroscopic interfaces (i.e., sand and plastic/glass) at high or low RH. Cell survival was conversely inhibited at high RH and promoted at low RH, irrespective of surface hygroscopicity. Based on this demonstration of metabolic persistence and survival inhibition at high RH, it was proposed that biofilm metabolic rates might inversely influence whole-biofilm resilience, with ‘resilience’ defined in this study as a biofilm’s capacity to recover from desiccation. The concept of whole-biofilm resilience being promoted by oligotrophy was supported in desiccation-tolerant Arthrobacter spp. biofilms, but not in desiccation-sensitive Pseudomonas aeruginosa biofilms. The ability of microbes to interact with surfaces to harness water vapor during desiccation was demonstrated, and potentially to harness oligotrophy (the most ubiquitous natural condition facing microbes) for adaptation to desiccation. PMID:27746774

  8. Improving adherence to safe prescription guidelines for Dapsone: harnessing an enhanced electronic medical records system and a team approach.

    Science.gov (United States)

    Neoh, Ching Yin; Tian, Elizabeth A L; Choi, Cindy; Leong, Hui Hui; Tan, Audrey W H; Chan, Roy K W

    2012-03-01

    Dapsone is a commonly prescribed medication in dermatological practice. Its use is associated with a broad spectrum of adverse effects. Careful selection and monitoring of patients on dapsone are paramount in the prevention and early recognition of adverse effects. We designed a risk-management program for dapsone at National Skin Centre, Singapore, enhancing an existing electronic medical records system and harnessing a team approach involving the nurses. This includes the performance of key laboratory tests before and after starting dapsone, ensuring adequate counseling before starting dapsone and screening for adverse effects using a questionnaire every visit. This system of dapsone prescription efficiently improved the adherence to safe prescription and monitoring guidelines. Average adherence rates for key safety parameters improved from 61.4% pre-implementation to 95.3% at six months and were sustained at 12 months at 91.3%. Percentage of follow-up cases in which all three key monitoring parameters were fulfilled increased from 9.5% to 79.6% (p=0.0001) after 12months. The percentage of new patients in which all four key monitoring parameters were met increased from 50% to 80%. It was not statistically significant possibly because of small patient numbers. This project has also translated into enhanced patient safety with dapsone dosages adjusted in 17 patients who experienced mild adverse effects. No severe adverse effects to dapsone were observed in the 12-month period. This example of risk management for dapsone may serve as a model for institutions looking at harnessing information technology and a team approach for safer prescription of high-alert medications. © 2012 The International Society of Dermatology.

  9. [Social marketers' lack of success in using CSM discipline to harness commercial resources and increase contraceptive prevalence].

    Science.gov (United States)

    Davies, J

    1984-01-01

    Social marketers have certainly shown that the CSM discipline can quickly and cost-effectively harness commercial resources to increase contraceptive prevalence. But why hasn't the social marketing idea caught on in more countries? According to Social Marketing Forum, only a tiny number of countries have active programs after more than a decade of effort. The most likely reason for this lack of success is doubt and fear on the part of both developing countries' officials and donor agencies about allowing marketing enthusiasts--with our very noticeable advertising methods--to join the family planning fight. And what has our answer been? Usually, a head-on retort such as, "But look at all the condoms we've sold]" And that gets us nowhere, because the successful peddling of 1 not-so-impressive method doesn't begin to balance the fears of a possible backlash that brash condom promotions could bring down on official heads. The lesson we should be learning is that social marketers possess to narrow an outlook and promote a small range of products that don't enthuse decisionmakers. Hence, we are often perceived as condom salesmen--and not much more. What should we be doing? We should be selling the idea of using private sector experience to assist national development. That means social development, particularly improved health, family planning and women's education. These 3 activities have proven effective in reducing fertility and would add up to a marketing opportunity--a longterm challenge that should enthuse dicisionmakers, private sector entrepreneurs and donor agencies alike. Further, our model--the commercial sector--is renowned for branching out and secceeding in a broad range of endeavors. As an example, Procter and Gamble uses separate divisions to market different products. Initiatives already exist in many countries to harness the private sector as a development tool. Social marketers should be leading the initiative--and benefitting from it, too. full text

  10. IL-15 Harnesses Pro-inflammatory Function of TEMRA CD8 in Kidney-Transplant Recipients

    Directory of Open Access Journals (Sweden)

    Gaëlle Tilly

    2017-06-01

    Full Text Available The involvement of TEMRA CD8 is evident in a large array of immunological conditions ranging from auto- to allo-immunity. Nevertheless, the factors leading to their accumulation and activation remain ill-defined and, efficient therapeutics to control their inflammatory response is lacking. Here, we show that IL-15-stimulated TEMRA from kidney-transplant (KT recipients promote inflammation by inducing the expression of CX3CL1 by endothelial cells in an IFN-γ- and TNF-α-dependent manner. The responsiveness of TEMRA to IL-15 is not restricted to chronic stimulation, as TEMRA from healthy volunteers respond earlier and faster when compared to effector memory (EM. IL-15 induces antiapoptotic signals and promotes proliferation dependent of PI3K/Akt, MAPK, and ERK pathways. Without ex vivo stimulation, TEMRA cells are metabolically more active than naive and EM, as shown by their high ATP reservoir and a high expression of genes involved in glycolysis, glutaminolysis, and the Pentose Phosphate Pathway. Upon stimulation, TEMRA adapt their metabolism by sustaining an increased mitochondrial respiration and glycolysis. Finally, we show that the inhibition of glycolysis is highly effective in preventing endothelial inflammation induced by TEMRA from KT recipients. Together, our findings highlight the metabolic fitness that tightly regulates the immune function of TEMRA in physiological and pathogenic situations.

  11. Harnessing the p53-PUMA Axis to Overcome DNA Damage Resistance in Renal Cell Carcinoma

    Directory of Open Access Journals (Sweden)

    Xiaoguang Zhou

    2014-12-01

    Full Text Available Resistance to DNA damage–induced apoptosis is a hallmark of cancer and a major cause of treatment failure and lethal disease outcome. A tumor entity that is largely resistant to DNA-damaging therapies including chemo- or radiotherapy is renal cell carcinoma (RCC. This study was designed to explore the underlying molecular mechanisms of DNA damage resistance in RCC to develop strategies to resensitize tumor cells to DNA damage–induced apoptosis. Here, we show that apoptosis-resistant RCC cells have a disconnect between activation of p53 and upregulation of the downstream proapoptotic protein p53 upregulated modulator of apoptosis (PUMA. We demonstrate that this disconnect is not caused by gene-specific repression through CCCTC-binding factor (CTCF but instead by aberrant chromatin compaction. Treatment with an HDAC inhibitor was found to effectively reactivate PUMA expression on the mRNA and protein level and to revert resistance to DNA damage–induced cell death. Ectopic expression of PUMA was found to resensitize a panel of RCC cell lines to four different DNA-damaging agents tested. Remarkably, all RCC cell lines analyzed were wild-type for p53, and a knockdown was likewise able to sensitize RCC cells to acute genotoxic stress. Taken together, our results indicate that DNA damage resistance in RCC is reversible, involves the p53-PUMA axis, and is potentially targetable to improve the oncological outcomes of RCC patients.

  12. Loss of HSulf-1 expression enhances tumorigenicity by inhibiting Bim expression in ovarian cancer.

    Science.gov (United States)

    He, Xiaoping; Khurana, Ashwani; Roy, Debarshi; Kaufmann, Scott; Shridhar, Viji

    2014-10-15

    The expression of human Sulfatase1 (HSulf-1) is downregulated in the majority of primary ovarian cancer tumors, but the functional consequence of this downregulation remains unclear. Using two different shRNAs (Sh1 and Sh2), HSulf-1 expression was stably downregulated in ovarian cancer OV202 cells. We found that HSulf-1-deficient OV202 Sh1 and Sh2 cells formed colonies in soft agar. In contrast, nontargeting control (NTC) shRNA-transduced OV202 cells did not form any colonies. Moreover, subcutaneous injection of OV202 HSulf-1-deficient cells resulted in tumor formation in nude mice, whereas OV202 NTC cells did not. Also, ectopic expression of HSulf-1 in ovarian cancer SKOV3 cells significantly suppressed tumor growth in nude mice. Here, we show that HSulf-1-deficient OV202 cells have markedly decreased expression of proapoptotic Bim protein, which can be rescued by restoring HSulf-1 expression in OV202 Sh1 cells. Enhanced expression of HSulf-1 in HSulf-1-deficient SKOV3 cells resulted in increased Bim expression. Decreased Bim levels after loss of HSulf-1 were due to increased p-ERK, because inhibition of ERK activity with PD98059 resulted in increased Bim expression. However, treatment with a PI3 kinase/AKT inhibitor, LY294002, failed to show any change in Bim protein level. Importantly, rescuing Bim expression in HSulf-1 knockdown cells significantly retarded tumor growth in nude mice. Collectively, these results suggest that loss of HSulf-1 expression promotes tumorigenicity in ovarian cancer through regulating Bim expression. © 2014 UICC.

  13. Generation of chickens expressing Cre recombinase.

    Science.gov (United States)

    Leighton, Philip A; Pedersen, Darlene; Ching, Kathryn; Collarini, Ellen J; Izquierdo, Shelley; Jacob, Roy; van de Lavoir, Marie-Cecile

    2016-10-01

    Cre recombinase has been extensively used for genome engineering in transgenic mice yet its use in other species has been more limited. Here we describe the generation of transgenic chickens expressing Cre recombinase. Green fluorescent protein (GFP)-positive chicken primordial germ cells were stably transfected with β-actin-Cre-recombinase using phiC31 integrase and transgenic chickens were generated. Cre recombinase activity was verified by mating Cre birds to birds carrying a floxed transgene. Floxed sequences were only excised in offspring from roosters that inherited the Cre recombinase but were excised in all offspring from hens carrying the Cre recombinase irrespective of the presence of the Cre transgene. The Cre recombinase transgenic birds were healthy and reproductively normal. The Cre and GFP genes in two of the lines were closely linked whereas the genes segregated independently in a third line. These founders allowed development of GFP-expressing and non-GFP-expressing Cre recombinase lines. These lines of birds create a myriad of opportunities to study developmentally-regulated and tissue-specific expression of transgenes in chickens.

  14. Harnessing QbD, Programming Languages, and Automation for Reproducible Biology.

    Science.gov (United States)

    Sadowski, Michael I; Grant, Chris; Fell, Tim S

    2016-03-01

    Building robust manufacturing processes from biological components is a task that is highly complex and requires sophisticated tools to describe processes, inputs, and measurements and administrate management of knowledge, data, and materials. We argue that for bioengineering to fully access biological potential, it will require application of statistically designed experiments to derive detailed empirical models of underlying systems. This requires execution of large-scale structured experimentation for which laboratory automation is necessary. This requires development of expressive, high-level languages that allow reusability of protocols, characterization of their reliability, and a change in focus from implementation details to functional properties. We review recent developments in these areas and identify what we believe is an exciting trend that promises to revolutionize biotechnology. Copyright © 2015 The Authors. Published by Elsevier Ltd.. All rights reserved.

  15. Enigma homolog 1 promotes myogenic gene expression and differentiation of C2C12 cells.

    Science.gov (United States)

    Ito, Jumpei; Takita, Masatoshi; Takimoto, Koichi; Maturana, Andrés D

    2013-06-07

    The Enigma homolog (ENH) gene generates several splicing variants. The initially identified ENH1 possesses one PDZ and three LIM domains, whereas ENH2~4 lack the latter domains. The splicing switch from ENH1 to LIM-less ENHs occurs during development/maturation of skeletal and heart muscles. We examined for the roles of ENH splicing variants in muscle differentiation using C2C12 cells. Cells stably expressing ENH1 exhibited significantly higher MyoD and myogenin mRNA levels before differentiation and after 5 days in low serum-differentiating medium than mock-transfected cells. ENH1-stable transformants also retained the ability to exhibit elongated morphology with well-extended actin fibers following differentiation. In contrast, cells stably expressing ENH3 or ENH4 did not show myotube-like morphology or reorganization of actin fibers following culture in the differentiating medium. Transient overexpression of ENH1 using adenovirus supported the increased expression of muscle marker mRNAs and the formation of well-organized stress fibers, whereas ENH4 overexpression prevented these morphological changes. Furthermore, specific suppression of ENH1 expression by RNAi caused a significant reduction in MyoD mRNA level and blocked the morphological changes. These results suggest that ENH1 with multiple protein-protein interaction modules is essential for differentiation of striated muscles, whereas ectopic expression of LIM-less ENH disrupts normal muscle differentiation. Copyright © 2013 Elsevier Inc. All rights reserved.

  16. Harness: Heterogeneous Adaptable Reconfigurable Networked Systems -- U.S. Department of Energy Grant DE-FG02-99ER25379 Final Project Report

    Energy Technology Data Exchange (ETDEWEB)

    Vaidy Sunderam

    2003-07-02

    Issues in reconfigurability and adaptability in heterogeneous distributed systems for high-performance computing are the focus of the work funded by this grant. Our efforts are part of an ongoing research project in metacomputing and are a follow on to the DOE funded PVM system that has witnessed over a decade of use at numerous institutions worldwide. The current project, termed Harness, investigates novel methodologies and tools for distributed metacomputing, focusing on dynamically reconfigurable software frameworks. During the first phase, we defined the metacomputing architecture embodied in Harness and developed prototype subsystems as proof of concept exercises. Subsequently, we designed and developed a complete software framework manifesting the Harness architecture, and also developed several tools and subsystems that demonstrated the viability and effectiveness of our proposed model for next generation metacomputing. We then used this substrate to emulate multiple programming environments on Harness, and conducted performance evaluation and tuning exercises. The main research results from these efforts include the establishment of software metacomputing systems as viable and cost-effective alternatives to MPPs; the demonstration of dynamic and reconfigurable platforms as effective methods of tailoring parallel computing environments; the development of methodologies to construct plugin modules for component-based distributed systems; contributions to performance modeling and optimization in emulated software environments; and software architectures for multi- and mixed-paradigm parallel distributed computing. Details and specifics on these and other results have been reported in numerous publications, and are manifested in software systems, all of which may be accessed at or via the website http://www.mathcs.emory.edu/harness/

  17. Effect of yeast TEL1 gene expression in A-T cells on chromosome aberration induced by radiation

    International Nuclear Information System (INIS)

    Cao Jianping; Zheng Siying; Zhu Wei; Luo Jialin; Zhou Xianfeng; Wang Mingming

    2004-01-01

    Objective: To analyze the effect of yeast TEL1 gene expression in human A-T (ataxia-telangiectasia) cells on chromosome aberration induced by radiation. Methods: Yeast TEL1 gene (TEL1rep), TEL1 fragment (deltaTELrep), pRep5 vector were stably transfected into A-T cells and pRep5 vector was stably transfected into GM639 cells. respectively. The stably transfected A-T and GM639 cells, namely, AT-TEL1, AT-deltaTEL1, AT-rep, GM-rep, were grown selectively in Dulbecco's minimal essential medium (DMEM) containing 100 μg/ml hygromycin, while non-transfected A-T and GM639 cells were grown in DMEM containing 15%220% FBS. 60 Co γ-rays were used to irradiate the above cells at exponential phase. The irradiation dose was 0, 1, 3 Gy, respectively. A hundred metaphases were analyzed for microscopically detectable chromosome type and chromatid type aberrations. Results: The chromosome numbers of all transfected, and non-transfected A-T, GM639 fobroblasts were aneuploid. Both spontaneous and radiation-induced chromosome aberration frequencies of A-T cells were higher than those of GM639 control cells. There is no significant difference in the spontaneous chromosome aberration between TEL1 gene stably transfected A-T cells (AT-TEL1) and non-transfected A-T cells (P>0.05). Chromosome aberration frequencies induced by radiation in AT-TEL1 cells were significantly lower than those of A-T cells (P<0.01). However, chromosome aberration frequencies induced by radiation in AT-deltaTEL1 and AT-rep cells were higher than those of A-T cells. Conclusion: The expression of yeast TEL1 gene in human A-T cells can significantly decrease the chromosome aberration frequencies induced by radiation. (authors)

  18. The hygiene theory harnessing helminths and their ova to treat autoimmunity.

    Science.gov (United States)

    Ben-Ami Shor, Dana; Harel, Michal; Eliakim, Rami; Shoenfeld, Yehuda

    2013-10-01

    The incidence of autoimmune diseases is increasing in Western countries, possibly due to the improved sanitary conditions and reduced exposure to infections in childhood (the hygiene hypothesis). There is an ongoing debate whether infection prevents or precipitates autoimmune diseases. Various helminths species used in several animal models were shown to limit inflammatory activity in a variety of diseases including inflammatory bowel disease, multiple sclerosis, type 1 diabetes, rheumatoid arthritis, and systemic lupus erythematosus. At present the scientific data is based mostly on experimental animal models; however, there is an increasing body of evidence in a number of clinical trials being conducted. Herein we review several clinical trials evaluating the anti-inflammatory effects of helminths and assessing their association with different autoimmune diseases, including inflammatory bowel disease, multiple sclerosis, and autoimmune liver diseases. We also describe the common pathways by which helminths induce immune modulation and the key changes observed in the host immune system following exposure to helminths. These common pathways include the inhibition of IFN-γ and IL-17 production, promotion of IL-4, IL-10 and TGF-β release, induction of CD4(+) T cell FoxP3(+) expression, and generation of regulatory macrophages, dendritic cells, and B cells. Helminths products are becoming significant candidates for anti-inflammatory agents in this context. However, further research is needed for synthetic analogues of helminths' potent products that mimic the parasite-mediated immunomodulation effect.

  19. Venous access and care: harnessing pragmatics in harm reduction for people who inject drugs.

    Science.gov (United States)

    Harris, Magdalena; Rhodes, Tim

    2012-06-01

    To explore the facilitators of long-term hepatitis C avoidance among people who inject drugs. We employed a qualitative life history design. Recruitment took place through low-threshold drug services and drug user networks in South East and North London. Participants were interviewed at the recruitment services or in their homes. The sample comprised 35 people who inject drugs, 20 of whom were hepatitis C antibody-negative. Participants' average injecting trajectory was 19 years (6-33), with 66% primarily injecting heroin, and 34% a crack and heroin mix. Nine (26%) of the sample were female and the average age was 39 years (23-53). Two interviews were conducted with each participant, with the second interview incorporating reference to a computer-constructed life history time-line. Interview accounts were audiorecorded, transcribed verbatim and analysed thematically. Hepatitis C risk awareness was recent and deprioritized by the majority of participants. The facilitation of venous access and care was an initial and enduring rationale for safe injecting practices. Difficult venous access resulted in increased contamination of injecting environments and transitions to femoral injecting. Participants expressed an unmet desire for non-judgemental venous access information and advice. Harm reduction interventions which attend to the immediate priorities of people who inject drugs, such as venous access and care, have the potential to re-engage individuals who are jaded or confused by hepatitis C prevention messages. © 2011 The Authors, Addiction © 2011 Society for the Study of Addiction.

  20. Harnessing what lies within: Programming immunity with biocompatible devices to treat human disease

    Science.gov (United States)

    Roberts, Reid Austin

    Advances in our mechanistic insight of cellular function and how this relates to host physiology have revealed a world which is intimately connected at the macro and micro level. Our increasing understanding of biology exemplifies this, where cells respond to environmental cues through interconnected networks of proteins which function as receptors and adaptors to elicit gene expression changes that drive appropriate cellular programs for a given stimulus. Consequently, our deeper molecular appreciation of host homeostasis implicates aberrations of these pathways in nearly all major human disease categories, including those of infectious, metabolic, neurologic, oncogenic, and autoimmune etiology. We have come to recognize the mammalian immune system as a common network hub among all these varied pathologies. As such, the major goal of this dissertation is to identify a platform to program immune responses in mammals so that we may enhance our ability to treat disease and improve health in the 21st century. Using advances in materials science, in particular a recently developed particle fabrication technology termed Particle Replication in Non-wetting Templates (PRINT), our studies systematically assess the murine and human immune response to precisely fabricated nano- and microscale particles composed of biodegradable and biocompatible materials. We then build on these findings and present particle design parameters to program a number of clinically attractive immune responses by targeting endogenous cellular signaling pathways. These include control of particle uptake through surface modification, design parameters that modulate the magnitude and kinetics of biological signaling dynamics that can be used to exacerbate or dampen inflammatory responses, as well as particle designs which may be of use in treating allergies and autoimmune disorders. In total, this dissertation provides evidence that rational design of biocompatible nano- and microparticles is a viable

  1. Harnessing genomics to improve health in the Eastern Mediterranean Region – an executive course in genomics policy

    Directory of Open Access Journals (Sweden)

    Singer Peter A

    2005-01-01

    Full Text Available Abstract Background While innovations in medicine, science and technology have resulted in improved health and quality of life for many people, the benefits of modern medicine continue to elude millions of people in many parts of the world. To assess the potential of genomics to address health needs in EMR, the World Health Organization's Eastern Mediterranean Regional Office and the University of Toronto Joint Centre for Bioethics jointly organized a Genomics and Public Health Policy Executive Course, held September 20th–23rd, 2003, in Muscat, Oman. The 4-day course was sponsored by WHO-EMRO with additional support from the Canadian Program in Genomics and Global Health. The overall objective of the course was to collectively explore how to best harness genomics to improve health in the region. This article presents the course findings and recommendations for genomics policy in EMR. Methods The course brought together senior representatives from academia, biotechnology companies, regulatory bodies, media, voluntary, and legal organizations to engage in discussion. Topics covered included scientific advances in genomics, followed by innovations in business models, public sector perspectives, ethics, legal issues and national innovation systems. Results A set of recommendations, summarized below, was formulated for the Regional Office, the Member States and for individuals. • Advocacy for genomics and biotechnology for political leadership; • Networking between member states to share information, expertise, training, and regional cooperation in biotechnology; coordination of national surveys for assessment of health biotechnology innovation systems, science capacity, government policies, legislation and regulations, intellectual property policies, private sector activity; • Creation in each member country of an effective National Body on genomics, biotechnology and health to: - formulate national biotechnology strategies - raise

  2. [Construction of acid-sensitive potassium channel-3 eukaryotic expression plasmid and its express in SH-SY5Y cells].

    Science.gov (United States)

    Wei, Lin-yu; Li, Xin-juan; Mei, Yi-wen; Wang, Guo-hong; Wang, Qi; Li, Dong-liang; Li, Chao-kun

    2015-05-01

    To construct the acid-sensitive potassium hannel-3(TASK3) eukaryotic expression plasmid and to establish a stable SH-SY5Y cell line expressing enhanced green fluorescent protein (EGFP)-tagged TASK3. TASK3 coding region was subcloned into pEGFP-N1 plasmid to construct a recombinant vector alled pEGFP-TASK3. The correct recombinant expressing plasmid was transfected with X-feet transfection reagent to SH-SY5Y cells. The cell line stably expressiing EGFP tagged-TASK3 gene was established by screening with antibiotic G418 and fluorescence microscope. The expression and localization of the EGFP tagged-TASK3 fusion protein was detected by Western blot and confocal microscope. Exposure of the SH-SY5Y cell line expressing stably TASK3-eGFP fusion proteins was exposed to different pH media (7.0, 6.7, 6.4, 6.1) for 24 h, the cell viability was assessed with cell counting Kit-8 (CCK-8). All the results of identification by PCR, digestion with restriction endonuclease and sequencing indicated that the recombinant eukaryotic expression plasmid pEGFP-TASK3 was constructed correctly. The stable SH-SY5Y cell line expressing EGFP tagged-TASK3 fusion protein was successfully established. Exposure of the wild type SH-SY5Y cells and the stable SH-SY5Y-GFP tag-TASK3 cell line to different pH media (7.0, 6.7, 6.4, 6.1) for 24 h, the cell viability of two group cells significantly reduced with pH declining, and the difference was statistically significant (P cells, the cell viability of stable SH-SYSY-GFP tag-TASK3 cell line increased significantly with the same pH media, and the difference was statistically significant (P eukaryotic expression vector pEGFP-TASK3 is successfully constructed and the cell line stably expressing TASK3-eGFP fusion is established which is important for their fundamental research and potential applications.

  3. Deregulation of microRNA expression in thyroid neoplasias.

    Science.gov (United States)

    Pallante, Pierlorenzo; Battista, Sabrina; Pierantoni, Giovanna Maria; Fusco, Alfredo

    2014-02-01

    MicroRNAs (miRNAs) have emerged as a class of powerful gene expression regulators. Acting at the post-transcriptional level, miRNAs modulate the expression of at least one-third of the mRNAs that are encoded by the human genome. The expression of a single gene can be regulated by several miRNAs, and every miRNA has more than one target gene. Thus, the miRNA regulatory circuit, which affects essential cellular functions, is of enormous complexity. Moreover, a fundamental role for miRNAs has been determined in the onset and progression of human cancers. Here, we summarize the main alterations in miRNA expression that have been identified in thyroid neoplasias and examine the mechanisms through which miRNA deregulation might promote thyroid cell transformation. We also discuss how the emerging knowledge on miRNA deregulation could be harnessed for the diagnosis and treatment of thyroid neoplasias.

  4. Evaluation of reference genes for gene expression analysis using quantitative RT-PCR in Azospirillum brasilense.

    Directory of Open Access Journals (Sweden)

    Mary McMillan

    Full Text Available Azospirillum brasilense is a nitrogen fixing bacterium that has been shown to have various beneficial effects on plant growth and yield. Under normal conditions A. brasilense exists in a motile flagellated form, which, under starvation or stress conditions, can undergo differentiation into an encapsulated, cyst-like form. Quantitative RT-PCR can be used to analyse changes in gene expression during this differentiation process. The accuracy of quantification of mRNA levels by qRT-PCR relies on the normalisation of data against stably expressed reference genes. No suitable set of reference genes has yet been described for A. brasilense. Here we evaluated the expression of ten candidate reference genes (16S rRNA, gapB, glyA, gyrA, proC, pykA, recA, recF, rpoD, and tpiA in wild-type and mutant A. brasilense strains under different culture conditions, including conditions that induce differentiation. Analysis with the software programs BestKeeper, NormFinder and GeNorm indicated that gyrA, glyA and recA are the most stably expressed reference genes in A. brasilense. The results also suggested that the use of two reference genes (gyrA and glyA is sufficient for effective normalisation of qRT-PCR data.

  5. Evaluation of reference genes for gene expression analysis using quantitative RT-PCR in Azospirillum brasilense.

    Science.gov (United States)

    McMillan, Mary; Pereg, Lily

    2014-01-01

    Azospirillum brasilense is a nitrogen fixing bacterium that has been shown to have various beneficial effects on plant growth and yield. Under normal conditions A. brasilense exists in a motile flagellated form, which, under starvation or stress conditions, can undergo differentiation into an encapsulated, cyst-like form. Quantitative RT-PCR can be used to analyse changes in gene expression during this differentiation process. The accuracy of quantification of mRNA levels by qRT-PCR relies on the normalisation of data against stably expressed reference genes. No suitable set of reference genes has yet been described for A. brasilense. Here we evaluated the expression of ten candidate reference genes (16S rRNA, gapB, glyA, gyrA, proC, pykA, recA, recF, rpoD, and tpiA) in wild-type and mutant A. brasilense strains under different culture conditions, including conditions that induce differentiation. Analysis with the software programs BestKeeper, NormFinder and GeNorm indicated that gyrA, glyA and recA are the most stably expressed reference genes in A. brasilense. The results also suggested that the use of two reference genes (gyrA and glyA) is sufficient for effective normalisation of qRT-PCR data.

  6. Codon-Optimized Luciola Italica Luciferase Variants for Mammalian Gene Expression in Culture and in Vivo

    Directory of Open Access Journals (Sweden)

    Casey A. Maguire

    2012-01-01

    Full Text Available Luciferases have proven to be useful tools in advancing our understanding of biologic processes. Having a multitude of bioluminescent reporters with different properties is highly desirable. We characterized codon-optimized thermostable green- and red-emitting luciferase variants from the Italian firefly Luciola italica for mammalian gene expression in culture and in vivo. Using lentivirus vectors to deliver and stably express these luciferases in mammalian cells, we showed that both variants displayed similar levels of activity and protein half-lives as well as similar light emission kinetics and higher stability compared to the North American firefly luciferase. Further, we characterized the red-shifted variant for in vivo bioluminescence imaging. Intramuscular injection of tumor cells stably expressing this variant into nude mice yielded a robust luciferase activity. Light emission peaked at 10 minutes post-D-luciferin injection and retained > 60% of signal at 1 hour. Similarly, luciferase activity from intracranially injected glioma cells expressing the red-shifted variant was readily detected and used as a marker to monitor tumor growth over time. Overall, our characterization of these codon-optimized luciferases lays the groundwork for their further use as bioluminescent reporters in mammalian cells.

  7. Ilfeld abduction orthosis is an effective second-line treatment after failure of Pavlik harness for infants with developmental dysplasia of the hip.

    Science.gov (United States)

    Sankar, Wudbhav N; Nduaguba, Afamefuna; Flynn, John M

    2015-02-18

    Closed reduction and spica casting is the most commonly recommended choice for infants with developmental dysplasia of the hip (DDH) for whom Pavlik harness treatment has failed, but it requires general anesthesia in addition to the challenges of spica cast care. The purposes of this study were to evaluate the effectiveness of Ilfeld bracing for infants for whom Pavlik harness treatment is unsuccessful and to compare these results with those for a similar cohort of patients directly undergoing closed reduction and spica casting. We reviewed the cases of a consecutive series of children with DDH who had failure of Pavlik harness treatment and were subsequently managed with Ilfeld bracing (the BR cohort) and compared this cohort with a similar historical group of infants who had failure of Pavlik harness treatment but had standard closed reduction and spica casting (the CR cohort). The cohorts were compared with respect to clinical and ultrasonographic data at the time of Pavlik discontinuation. At one year, the hip stability and acetabular index were assessed; the presence of osteonecrosis was graded according to the criteria described by Salter et al. Twenty-eight hips (nineteen infants) made up the BR cohort and twenty-two hips (sixteen infants) made up the CR cohort. Ultrasonographic indices (including the alpha angle and the percentage of femoral head coverage) were comparable between the two cohorts (p=0.66 and 0.19, respectively). Following treatment, a stable reduction was achieved in twenty-three (82%) of twenty-eight hips in the BR cohort compared with twenty (91%) of twenty-two hips in the CR cohort. At one year, acetabular indices were similar between both cohorts (mean and standard deviation, 27°±6° for the BR cohort versus 27°±5° for the CR cohort; p=0.62); however, osteonecrosis developed in three hips in the CR cohort compared with none in the BR cohort. In our series of infants with DDH for whom Pavlik harness treatment had failed, Ilfeld

  8. Efficient replication and expression of murine leukemia virus with major deletions in the enhancer region of U3

    DEFF Research Database (Denmark)

    Pedersen, K.; Lovmand, S.; Bonefeld-Jørgensen, Eva Cecilie

    1992-01-01

    of the viral RNA. Genetic tagging of the retrovirus with lacO facilitated the analysis. Among the individual mutated LTRs an over 100-fold difference in a transient expression assay was previously detected. This difference was not revealed in studies of viral replication in cell culture, where the expression......The effect of deletions within the enhancer region in the U3 part of the LTR derived from the murine retrovirus Akv was studied. The deletions were stably transmitted through normal virus replication as shown by sequence analysis of cloned polymerase chain reaction product of the cDNA copy...

  9. CoDiagnose: Interactive software to harness collaborative diagnoses and to increase diagnostic accuracy amongst junior physicians.

    Science.gov (United States)

    McComb, Steven; Bond, Raymond R

    2015-01-01

    Many studies have been conducted that highlight the prevalence of misdiagnoses within the medical profession and the statistically poor diagnostic accuracy rates associated with junior clinicians. There is evidence that crowdsourcing and multiple medical opinions can improve the veracity of a diagnosis. This paper proposes a cloud based prototype that makes use of interactive web technologies along with the concepts of crowdsourcing and e-learning to improve diagnostic accuracy rates amongst junior doctors. Existing approaches in this field are analysed to identify potential gaps and opportunities. Sample representations of both junior and senior clinicians are surveyed to establish the feasibility of adopting collaborative diagnostics. It is theorised that the prototype developed in this work will harness the experience and knowledge of expert clinicians. The prototype is a fully responsive web-based and feature-rich interactive system that has been developed using the latest Internet technologies. It has been evaluated using usability tests and a one month long trial involving a sample of end-users. Crowdsourcing is an innovative process in its infancy but the results being produced indicate a promising future for its use within medicine. The results offer an inclination that the prototype's usage could be attributed to improved diagnostic ability. However the findings need to be replicated in larger, independent samples.

  10. Harnessing the power of communities: Career networking strategies for bioscience PhD students and postdoctoral researchers.

    Science.gov (United States)

    Blackford, Sarah

    2018-02-09

    With an ever more competitive global labour market, coupled with an ever-increasing population of PhD-qualified graduates, the ability to communicate effectively and build strategic connections with others can be advantageous in the job-search process. Whether in pursuit of a tenure-track or non-academic position, many postdoctoral researchers and PhD students will benefit from networking as early as possible to enhance their career propsects. Sometimes viewed cynically as 'using people' or dismissed as 'the old boy network', the ability to make meaningful connections and build relationships can be more valuable than other job-related skills in order to gain entry to, and progress within, many professions.This mini-review highlights the positive influence of networking and how bioscience PhD students and postdoctoral researchers can harness the power of communities to achieve career success. It is argued that those who make connections and promote personal patronage through networking can gain an advantage over their contemporaries. A summary of key theories and research studies that underpin the practice of networking provide credence to these assertions, which are further substantiated with examples pertinent to the academic community. Although primarily focussed on the biosciences, much of the content is applicable to other scientists at a similar career stage. © FEMS 2018. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.

  11. Harnessing mode-selective nonlinear optics for on-chip multi-channel all-optical signal processing

    Directory of Open Access Journals (Sweden)

    Ming Ma

    2016-11-01

    Full Text Available All-optical signal processing based on nonlinear optical effects allows for the realization of important functions in telecommunications including wavelength conversion, optical multiplexing/demultiplexing, Fourier transformation, and regeneration, amongst others, on ultrafast time scales to support high data rate transmission. In integrated photonic subsystems, the majority of all-optical signal processing systems demonstrated to date typically process only a single channel at a time or perform a single processing function, which imposes a serious limitation on the functionality of integrated solutions. Here, we demonstrate how nonlinear optical effects can be harnessed in a mode-selective manner to perform simultaneous multi-channel (two and multi-functional optical signal processing (i.e., regenerative wavelength conversion in an integrated silicon photonic device. This approach, which can be scaled to a higher number of channels, opens up a new degree of freedom for performing a broad range of multi-channel nonlinear optical signal processing functions using a single integrated photonic device.

  12. Harnessing the Maltodextrin Transport Mechanism for Targeted Bacterial Imaging: Structural Requirements for Improved in vivo Stability in Tracer Design.

    Science.gov (United States)

    Axer, Alexander; Hermann, Sven; Kehr, Gerald; Clases, David; Karst, Uwe; Fischer-Riepe, Lena; Roth, Johannes; Fobker, Manfred; Schäfers, Michael; Gilmour, Ryan; Faust, Andreas

    2018-02-06

    Diagnosis and localization of bacterial infections remains a significant clinical challenge. Harnessing bacteria-specific metabolic pathways, such as the maltodextrin transport mechanism, may allow specific localization and imaging of small or hidden colonies. This requires that the intrabacterial tracer accumulation provided by the transporter is matched by high serum stability of the tracer molecule. Herein, radiolabeled maltodextrins of varying chain lengths and with free nonreducing/reducing ends are reported and their behavior against starch-degrading enzymes in the blood, which compromise their serum stability, is evaluated. Successful single-photon emission computed tomography (SPECT) imaging is shown in a footpad infection model in vivo by using the newly developed model tracer, [ 99m Tc]MB1143, and the signal is compared with that of 18 F-fluorodeoxyglucose positron emission tomography ([ 18 F]FDG-PET) as a nonbacterial specific marker for inflammation. Although the [ 99m Tc]MB1143 imaging signal is highly specific, it is low, most probably due to insufficient serum stability of the tracer. A series of stability tests with different 18 F-labeled maltodextrins finally yielded clear structural guidelines regarding substitution patterns and chain lengths of maltodextrin-based tracers for nuclear imaging of bacterial infections. © 2018 Wiley-VCH Verlag GmbH & Co. KGaA, Weinheim.

  13. An isotropic suspension system for a biaxial accelerometer using electroplated thick metal with a HAR SU-8 mold

    International Nuclear Information System (INIS)

    Lee, Jin Seung; Lee, Seung S

    2008-01-01

    In this paper, a novel approach is developed to design an isotropic suspension system using thick metal freestanding micro-structures combining bulk micro-machining with electroplating based on a HAR SU-8 mold. An omega-shape isotropic suspension system composed of circular curved beams that have free switching of imaginary boundary conditions is proposed. This novel isotropic suspension design is not affected by geometric dimensional parameters and always achieves matching stiffness along the principle axes of elasticity. Using the finite element method, the isotropic suspension system was compared with an S-shaped meandering suspension system. In order to realize the suggested isotropic suspension system, a cost-effective fabrication process using electroplating with the SU-8 mold was developed to avoid expensive equipment and materials such as deep reactive-ion etching (DRIE) or a silicon-on-insulator (SOI) wafer. The fabricated isotropic suspension system was verified by electromagnetic actuation experiments. Finally, a biaxial accelerometer with isotropic suspension system was realized and tested using a vibration generator system. The proposed isotropic suspension system and the modified surface micro-machining technique based on electroplating with an SU-8 mold can contribute towards minimizing the system size, simplifying the system configuration, reducing the system price of and facilitating mass production of various types of low-cost sensors and actuators

  14. Mechanics of hip dysplasia reductions in infants using the Pavlik harness: a physics-based computational model.

    Science.gov (United States)

    Ardila, Orlando J; Divo, Eduardo A; Moslehy, Faissal A; Rab, George T; Kassab, Alain J; Price, Charles T

    2013-05-31

    Biomechanical factors influencing the reduction of dislocated hips with the Pavlik harness in patients of Developmental Dysplasia of the Hip (DDH) were studied using a three-dimensional computer model simulating hip reduction dynamics in (1) subluxated and (2) fully dislocated hip joints. Five hip adductor muscles were identified as key mediators of DDH prognosis, and the non-dimensional force contribution of each in the direction necessary to achieve concentric hip reductions was determined. Results point to the adductor muscles as mediators of subluxated hip reductions, as their mechanical action is a function of the degree of hip dislocation. For subluxated hips in abduction and flexion, the Pectineus, Adductor Brevis, Adductor Longus, and proximal Adductor Magnus contribute positively to reduction, while the rest of the Adductor Magnus contributes negatively. In full dislocations all muscles contribute detrimentally to reduction, elucidating the need for traction to reduce Graf IV type dislocations. Reduction of dysplastic hips was found to occur in two distinct phases: (a) release phase and (b) reduction phase. Copyright © 2013 Elsevier Ltd. All rights reserved.

  15. Harnessing the private health sector by using prices as a policy instrument: Lessons learned from South Africa.

    Science.gov (United States)

    Barber, Sarah L; Kumar, Ankit; Roubal, Tomas; Colombo, Francesca; Lorenzoni, Luca

    2018-03-29

    Governments frequently draw upon the private health care sector to promote sustainability, optimal use of resources, and increased choice. In doing so, policy-makers face the challenge of harnessing resources while grappling with the market failures and equity concerns associated with private financing of health care. The growth of the private health sector in South Africa has fundamentally changed the structure of health care delivery. A mutually reinforcing ecosystem of private health insurers, private hospitals and specialists has grown to account for almost half of the country's spending on health care, despite only serving 16% of the population with the capacity to pay. Following years of consolidation among private hospital groups and insurance schemes, and after successive failures at establishing credible price benchmarks, South Africa's private hospitals charge prices comparable with countries that are considerably richer. This compromises the affordability of a broad-based expansion in health care for the population. The South African example demonstrates that prices can be part of a structure that perpetuates inequalities in access to health care resources. The lesson for other countries is the importance of norms and institutions that uphold price schedules in high-income countries. Efforts to compromise or liberalize price setting should be undertaken with a healthy degree of caution. Copyright © 2018 Elsevier B.V. All rights reserved.

  16. Harnessing endogenous miRNAs to control virus tissue tropism as a strategy for developing attenuated virus vaccines.

    Science.gov (United States)

    Barnes, Dwight; Kunitomi, Mark; Vignuzzi, Marco; Saksela, Kalle; Andino, Raul

    2008-09-11

    Live attenuated vaccines remain the safest, most cost-effective intervention against viral infections. Because live vaccine strains are generated empirically and the basis for attenuation is usually ill defined, many important viruses lack an efficient live vaccine. Here, we present a general strategy for the rational design of safe and effective live vaccines that harnesses the microRNA-based gene-silencing machinery to control viral replication. Using poliovirus as a model, we demonstrate that insertion of small miRNA homology sequences into a viral genome can restrict its tissue tropism, thereby preventing pathogenicity and yielding an attenuated viral strain. Poliovirus strains engineered to become targets of neuronal-specific miRNAs lost their ability to replicate in the central nervous system, leading to significant attenuation of neurovirulence in infected animals. Importantly, these viruses retained the ability to replicate in nonneuronal tissues. As a result, these engineered miRNA-regulated viruses elicited strong protective immunity in mice without producing disease.

  17. Inspiration in the harness of daily labor. Darwin, botany, and the triumph of evolution, 1859-1868.

    Science.gov (United States)

    Bellon, Richard

    2011-09-01

    Charles Darwin hoped that a large body of working naturalists would embrace evolution after the Origin of Species appeared in late 1859. He was disappointed. His evolutionary ideas at first made painfully little progress in the scientific community. But by 1863 the tide had turned dramatically, and within five years evolution became scientific orthodoxy in Britain. The Origin's reception followed this peculiar trajectory because Darwin had not initially tied its theory to productive original scientific investigation, which left him vulnerable to charges of reckless speculation. The debate changed with his successful application of evolution to original problems, most notably orchid fertilization, the subject of a well-received book in 1862. Most of Darwin's colleagues found the argument of the Origin convincing when they realized that it functioned productively in the day-to-day work of science-and not before. The conceptual force of the Origin, however outwardly persuasive, acquired full scientific legitimacy only when placed "in the harness of daily labour".

  18. Upgrading Wood-Based Industries: Harnessing the Social Network of Small-Scale Furniture Producers and Their Institutions

    Directory of Open Access Journals (Sweden)

    Melati ,

    2011-05-01

    Full Text Available Furniture is a major export commodity in Indonesia with a total value of USD 1.96 million in 2007.  Jepara District is one of the key location for wood furniture production with 15,271 furniture related business units employing 176,469 workers.  However, inefficiencies and power imbalances throughout the furniture value chain have resulted in overharvesting and uneven distribution of gains among the industry’s actors.  In contrast to price-setting international furniture retailers, small-scale producers enjoy the least value from their products.  In order to increase added value and competitiveness, small-scale furniture producers have made efforts to upgrade by harnessing their social network and institutions.  This paper describes small-scale furniture producers’ efforts to upgrade by utilising their social network and institutions in Jepara.  Data was collected through in-depth interviews with members of the small-scale furniture producers’ association.  The research provides insight into the nature of social networks and information flow and develops future scenarios to upgrade.  The scenarios will not only benefit the furniture industry in Jepara, but may also be adopted for similar industries throughout Indonesia and the world, and potentially improve many people’s economies and livelihoods.Keywords: wood-based industry, furniture, small-scale, social network, institution

  19. Harnessing endogenous miRNAs to control virus tissue tropism as a strategy for developing attenuated virus vaccines

    Science.gov (United States)

    Barnes, Dwight; Kunitomi, Mark; Vignuzzi, Marco; Saksela, Kalle; Andino, Raul

    2008-01-01

    Live, attenuated vaccines remain the safest, most cost-effective intervention against viral infections. Because live vaccine strains are generated empirically and the basis for attenuation is usually ill defined, many important viruses lack an efficient live vaccine. Here, we present a general strategy for the rational design of safe and effective live vaccines that harnesses the microRNA-based gene silencing machinery to control viral replication. Using poliovirus as a model, we demonstrate that insertion of small miRNA homology sequences into a viral genome can restrict its tissue tropism, thereby preventing pathogenicity and yielding an attenuated viral strain. Poliovirus strains engineered to become targets of neuronal-specific miRNAs lost their ability to replicate in the central nervous system, leading to significant attenuation of neurovirulence in infected animals. Importantly, these viruses retained the ability to replicate in non-neuronal tissues. As a result, these engineered miRNA-regulated viruses elicited strong protective immunity in mice without producing disease. PMID:18779050

  20. Biphasic action of cyclic adenosine 3',5'- monophosphate in gonadotropin-releasing hormone (GnRH) analog-stimulated hormone release from GH3 cells stably transfected with GnRH receptor complementary deoxyribonucleic acid.

    Science.gov (United States)

    Stanislaus, D; Arora, V; Awara, W M; Conn, P M

    1996-03-01

    GH3 cells are a PRL-secreting adenoma cell line derived from pituitary lactotropes. These cells have been stably transfected with rat GnRH receptor complementary DNA to produce four cell lines: GGH(3)1', GGH(3)2', GGH(3)6', and GGH(3)12'. In response to either GnRH or Buserelin (a metabolically stable GnRH agonist), these cell lines synthesize PRL in a cAMP-dependent manner. Only GGH(3)6' cells desensitize in response to persistent treatment with 10(-7) g/ml Buserelin. GGH(3)1', GGH(3)2', and GGH(3)12' cells, however, can be made refractory to Buserelin stimulation by raising cAMP levels either by the addition of (Bu)2cAMP to the medium or by treatment with cholera toxin. In GGH(3) cells, low levels of cAMP fulfill the requirements for a second messenger, whereas higher levels appear to mediate the development of desensitization. The observation that in GGH(3)6' cells, cAMP production persists after the onset of desensitization is consistent with the view that the mechanism responsible for desensitization is distal to the production of cAMP. Moreover, the absence of any significant difference in the amount of cAMP produced per cell in GGH(3)2', GGH(3)6', or GGH(3)12' cells suggests that elevated cAMP production per cell does not explain the development of desensitization in GGH(3)6' cells. We suggest that Buserelin-stimulated PRL synthesis in GGH(3)6' cells is mediated by a different cAMP-dependent protein kinase pool(s) than that in nondesensitizing GGH(3) cells. Such a protein kinase A pool(s) may be more susceptible to degradation via cAMP-mediated mechanisms than the protein kinase pools mediating the Buserelin response in nondesensitizing GGH(3) cells. A similar mechanism has been reported in other systems.

  1. Molekulare Klonierung, stabile Transfektion und funktionelle Expression der murinen Chemokinrezeptoren Ccr2 und Ccr5

    OpenAIRE

    Simonis, Christopher

    2009-01-01

    The two chemokine receptors CCR2 and CCR5 play important roles in the recruitment and activation of monocytes/macrophages and T-lymphocytes at sites of infection and inflammation. To further examine their function, I cloned the two murine chemokine receptors Ccr2 and Ccr5 from genomic mouse DNA by a PCR-based cloning strategy and functionally expressed them in stably transfected CHO-cells. These cells were used to generate the first monoclonal antibodies against Ccr2 and Ccr5.

  2. Det er en kirkelig leder jeg er. Hvordan og på hvilken måte har kirkevergens lederrolle i Den norske kirke utviklet seg gjennom de siste tjue årene?

    OpenAIRE

    Fiske, Mildrid

    2017-01-01

    Tema for denne masteroppgaven er hvordan og på hvilken måte kirkevergens lederolle har utviklet seg gjennom de siste tjue årene. Kirkevergene er virksomhetsledere i Den norske kirke. Formålet med studien har vært å undersøke hvilke erfaringer kirkevergene selv har gjort som ledere i Den norske kirke. I kirkeloven av 1997 ble det lovbestemt at det skulle være en virksomhetsleder i alle kirkelige fellesråd. Kirkevergevergestillingen ble da en ny lederstilling i Den norske kirke med et manda...

  3. Så galt går det når du ikke har styr på din SEO-partner

    DEFF Research Database (Denmark)

    Ivang, Reimer

    2014-01-01

    Interflora UK lå nummer et på Google på flere søgeord, men fra den ene dag til den anden havde Google fjernet dem. Så spørgsmålet er nu om Interflora UK ikke har styr på, hvad deres SEO partner foretager sig, om de var for grådige eller om de bare er blevet uretfærdig behandlet af Google....

  4. Vector Development for the Expression of Foreign Proteins in the Vaccine Strain Brucella abortus S19

    Science.gov (United States)

    Comerci, Diego J.; Pollevick, Guido D.; Vigliocco, Ana M.; Frasch, Alberto C. C.; Ugalde, Rodolfo A.

    1998-01-01

    A vector for the expression of foreign antigens in the vaccine strain Brucella abortus S19 was developed by using a DNA fragment containing the regulatory sequences and the signal peptide of the Brucella bcsp31 gene. This fragment was cloned in broad-host-range plasmid pBBR4MCS, resulting in plasmid pBEV. As a reporter protein, a repetitive antigen of Trypanosoma cruzi was used. The recombinant fusion protein is stably expressed and secreted into the Brucella periplasmic space, inducing a good antibody response against the T. cruzi antigen. The expression of the repetitive antigen in Brucella neither altered its growth pattern nor generated a toxic or lethal effect during experimental infection. The application of this strategy for the generation of live recombinant vaccines and the tagging of B. abortus S19 vaccine is discussed. This is the first time that a recombinant protein has been expressed in the periplasm of brucellae. PMID:9673273

  5. A comparative evaluation of methicillin-resistant staphylococci isolated from harness racing-horses, breeding mares and riding-horses in Italy.

    Science.gov (United States)

    Mallardo, Karina; Nizza, Sandra; Fiorito, Filomena; Pagnini, Ugo; De Martino, Luisa

    2013-03-01

    To investigate the prevalence of methicillin-resistant staphylococci (MRS) which is a potencial risk factor of transmission between animals and humans in different types of horses (harness racing-horses, breeding mares and riding-horses) and to compare the antimicrobial resistance of the isolates. A total of 191 healthy horses, housed at different locations of the Campania Region (Italy), were included in the study. Nasal swab samples were collected from each nostril of the horses. The mecA gene was detected by a nested PCR technique. Antibiotic susceptibility was tested for each isolate. MRS was isolated from nasal samples of 68/191 (35.6%; 95% CI: 28.9%-42.9%) healthy horses. All isolates were coagulase-negative with the exception of two coagulase-positive MRS strains, identified as Staphylococcus aureus and Staphylococcus pseudintermedius, 2/83 (2.4%; 95% CI: 0.4%-9.2%). Interestingly, both coagulase-positive MRS isolates were from harness racing-horses. These horses also presented a significantly higher positivity for MRS (53.3%; 95% CI: 40.1%-66.1%) than the breeding mares and riding-horses groups. Antibiotic susceptibility testing showed difference between isolates due to different origins except for an almost common high resistance to aminopenicillins, such as ampicillin and amoxicillin. It can be concluded that harness racing-horses may act as a significant reservoir of MRS as compared to breeding mares and riding-horses.

  6. The impact of intragenic CpG content on gene expression.

    Science.gov (United States)

    Bauer, Asli Petra; Leikam, Doris; Krinner, Simone; Notka, Frank; Ludwig, Christine; Längst, Gernot; Wagner, Ralf

    2010-07-01

    The development of vaccine components or recombinant therapeutics critically depends on sustained expression of the corresponding transgene. This study aimed to determine the contribution of intragenic CpG content to expression efficiency in transiently and stably transfected mammalian cells. Based upon a humanized version of green fluorescent protein (GFP) containing 60 CpGs within its coding sequence, a CpG-depleted variant of the GFP reporter was established by carefully modulating the codon usage. Interestingly, GFP reporter activity and detectable protein amounts in stably transfected CHO and 293 cells were significantly decreased upon CpG depletion and independent from promoter usage (CMV, EF1 alpha). The reduction in protein expression associated with CpG depletion was likewise observed for other unrelated reporter genes and was clearly reflected by a decline in mRNA copy numbers rather than translational efficiency. Moreover, decreased mRNA levels were neither due to nuclear export restrictions nor alternative splicing or mRNA instability. Rather, the intragenic CpG content influenced de novo transcriptional activity thus implying a common transcription-based mechanism of gene regulation via CpGs. Increased high CpG transcription correlated with changed nucleosomal positions in vitro albeit histone density at the two genes did not change in vivo as monitored by ChIP.

  7. Harnessing Fluorine-Sulfur Contacts and Multipolar Interactions for the Design of p53 Mutant Y220C Rescue Drugs.

    Science.gov (United States)

    Bauer, Matthias R; Jones, Rhiannon N; Baud, Matthias G J; Wilcken, Rainer; Boeckler, Frank M; Fersht, Alan R; Joerger, Andreas C; Spencer, John

    2016-08-19

    Many oncogenic mutants of the tumor suppressor p53 are conformationally unstable, including the frequently occurring Y220C mutant. We have previously developed several small-molecule stabilizers of this mutant. One of these molecules, PhiKan083, 1-(9-ethyl-9H-carbazole-3-yl)-N-methylmethanamine, binds to a mutation-induced surface crevice with a KD = 150 μM, thereby increasing the melting temperature of the protein and slowing its rate of aggregation. Incorporation of fluorine atoms into small molecule ligands can substantially improve binding affinity to their protein targets. We have, therefore, harnessed fluorine-protein interactions to improve the affinity of this ligand. Step-wise introduction of fluorines at the carbazole ethyl anchor, which is deeply buried within the binding site in the Y220C-PhiKan083 complex, led to a 5-fold increase in affinity for a 2,2,2-trifluoroethyl anchor (ligand efficiency of 0.3 kcal mol(-1) atom(-1)). High-resolution crystal structures of the Y220C-ligand complexes combined with quantum chemical calculations revealed favorable interactions of the fluorines with protein backbone carbonyl groups (Leu145 and Trp146) and the sulfur of Cys220 at the mutation site. Affinity gains were, however, only achieved upon trifluorination, despite favorable interactions of the mono- and difluorinated anchors with the binding pocket, indicating a trade-off between energetically favorable protein-fluorine interactions and increased desolvation penalties. Taken together, the optimized carbazole scaffold provides a promising starting point for the development of high-affinity ligands to reactivate the tumor suppressor function of the p53 mutant Y220C in cancer cells.

  8. Harnessing sensor and information/communication technologies to revolutionize how environmental data are collected and integrated to protect public health

    International Nuclear Information System (INIS)

    MacDonell, Margaret M.; Raymond, Michelle; Chang, Young-Soo; Armbruster, Walter J.

    2013-01-01

    Monitoring the levels of pollutants in ambient air has long been the responsibility of environmental agencies, and traditional programs have relied on a relatively small number of fixed stations that are expensive to install and operate. The spatial coverage of pollutant data ranges from limited in many urban areas to very sparse in smaller suburban and rural areas. Increasingly, the latter are being affected by new pollutant sources such as backyard drilling systems for energy development, and concentrated animal feeding operations and biodiesel production facilities on former agricultural lands. Regional measurement stations are not able to inform local communities about the types and concentrations of pollutants in their ambient and indoor air. Meanwhile, epidemiology studies and clinical data continue to indicate a link between air pollutants and adverse health effects, including respiratory and cardiovascular disease. With asthma on the rise and heart disease among the leading killers, the public has become increasingly concerned about knowing what their personal exposure levels are so they can determine appropriate measures to protect their health. To address this long-standing need, the U.S. Environmental Protection Agency (EPA) has embarked upon an innovative program to improve the understanding of air pollution at the community and neighborhood scales. This program harnesses striking advances in sensor technology, mobile applications, and environmental informatics, with citizens playing a key role. The goal is to facilitate the development and widespread use of inexpensive mobile sensors, for widespread collection and integration of air quality data by citizens across the nation (and world), to help guide environmental health protection programs. (orig.)

  9. Harnessing data structure for recovery of randomly missing structural vibration responses time history: Sparse representation versus low-rank structure

    Science.gov (United States)

    Yang, Yongchao; Nagarajaiah, Satish

    2016-06-01

    Randomly missing data of structural vibration responses time history often occurs in structural dynamics and health monitoring. For example, structural vibration responses are often corrupted by outliers or erroneous measurements due to sensor malfunction; in wireless sensing platforms, data loss during wireless communication is a common issue. Besides, to alleviate the wireless data sampling or communication burden, certain accounts of data are often discarded during sampling or before transmission. In these and other applications, recovery of the randomly missing structural vibration responses from the available, incomplete data, is essential for system identification and structural health monitoring; it is an ill-posed inverse problem, however. This paper explicitly harnesses the data structure itself-of the structural vibration responses-to address this (inverse) problem. What is relevant is an empirical, but often practically true, observation, that is, typically there are only few modes active in the structural vibration responses; hence a sparse representation (in frequency domain) of the single-channel data vector, or, a low-rank structure (by singular value decomposition) of the multi-channel data matrix. Exploiting such prior knowledge of data structure (intra-channel sparse or inter-channel low-rank), the new theories of ℓ1-minimization sparse recovery and nuclear-norm-minimization low-rank matrix completion enable recovery of the randomly missing or corrupted structural vibration response data. The performance of these two alternatives, in terms of recovery accuracy and computational time under different data missing rates, is investigated on a few structural vibration response data sets-the seismic responses of the super high-rise Canton Tower and the structural health monitoring accelerations of a real large-scale cable-stayed bridge. Encouraging results are obtained and the applicability and limitation of the presented methods are discussed.

  10. Harnessing sensor and information/communication technologies to revolutionize how environmental data are collected and integrated to protect public health

    Energy Technology Data Exchange (ETDEWEB)

    MacDonell, Margaret M.; Raymond, Michelle; Chang, Young-Soo [Argonne National Laboratory, Lemont, IL (United States); Armbruster, Walter J. [Farm Foundation, Darien, IL (United States)

    2013-07-01

    Monitoring the levels of pollutants in ambient air has long been the responsibility of environmental agencies, and traditional programs have relied on a relatively small number of fixed stations that are expensive to install and operate. The spatial coverage of pollutant data ranges from limited in many urban areas to very sparse in smaller suburban and rural areas. Increasingly, the latter are being affected by new pollutant sources such as backyard drilling systems for energy development, and concentrated animal feeding operations and biodiesel production facilities on former agricultural lands. Regional measurement stations are not able to inform local communities about the types and concentrations of pollutants in their ambient and indoor air. Meanwhile, epidemiology studies and clinical data continue to indicate a link between air pollutants and adverse health effects, including respiratory and cardiovascular disease. With asthma on the rise and heart disease among the leading killers, the public has become increasingly concerned about knowing what their personal exposure levels are so they can determine appropriate measures to protect their health. To address this long-standing need, the U.S. Environmental Protection Agency (EPA) has embarked upon an innovative program to improve the understanding of air pollution at the community and neighborhood scales. This program harnesses striking advances in sensor technology, mobile applications, and environmental informatics, with citizens playing a key role. The goal is to facilitate the development and widespread use of inexpensive mobile sensors, for widespread collection and integration of air quality data by citizens across the nation (and world), to help guide environmental health protection programs. (orig.)

  11. Harnessing the Risk-Related Data Supply Chain: An Information Architecture Approach to Enriching Human System Research and Operations Knowledge

    Science.gov (United States)

    Buquo, Lynn; Johnson-Throop, Kathy

    2010-01-01

    NASA's Human Research Program (HRP) and Space Life Sciences Directorate (SLSD), not unlike many NASA organizations today, struggle with the inherent inefficiencies caused by dependencies on heterogeneous data systems and silos of data and information spread across decentralized discipline domains. The capture of operational and research-based data/information (both in-flight and ground-based) in disparate IT systems impedes the extent to which that data/information can be efficiently and securely shared, analyzed, and enriched into knowledge that directly and more rapidly supports HRP's research-focused human system risk mitigation efforts and SLSD s operationally oriented risk management efforts. As a result, an integrated effort is underway to more fully understand and document how specific sets of risk-related data/information are generated and used and in what IT systems that data/information currently resides. By mapping the risk-related data flow from raw data to useable information and knowledge (think of it as the data supply chain), HRP and SLSD are building an information architecture plan to leverage their existing, shared IT infrastructure. In addition, it is important to create a centralized structured tool to represent risks including attributes such as likelihood, consequence, contributing factors, and the evidence supporting the information in all these fields. Representing the risks in this way enables reasoning about the risks, e.g. revisiting a risk assessment when a mitigation strategy is unavailable, updating a risk assessment when new information becomes available, etc. Such a system also provides a concise way to communicate the risks both within the organization as well as with collaborators. Understanding and, hence, harnessing the human system risk-related data supply chain enhances both organizations' abilities to securely collect, integrate, and share data assets that improve human system research and operations.

  12. Harnessing Online Peer Education (HOPE): integrating C-POL and social media to train peer leaders in HIV prevention.

    Science.gov (United States)

    Jaganath, Devan; Gill, Harkiran K; Cohen, Adam Carl; Young, Sean D

    2012-01-01

    Novel methods, such as Internet-based interventions, are needed to combat the spread of HIV. While past initiatives have used the Internet to promote HIV prevention, the growing popularity, decreasing digital divide, and multi-functionality of social networking sites, such as Facebook, make this an ideal time to develop innovative ways to use online social networking sites to scale HIV prevention interventions among high-risk groups. The UCLA Harnessing Online Peer Education study is a longitudinal experimental study to evaluate the feasibility, acceptability, and preliminary effectiveness of using social media for peer-led HIV prevention, specifically among African American and Latino Men who have Sex with Men (MSM). No curriculum currently exists to train peer leaders in delivering culturally aware HIV prevention messages using social media. Training was created that adapted the Community Popular Opinion Leader (C-POL) model, for use on social networking sites. Peer leaders are recruited who represent the target population and have experience with both social media and community outreach. The curriculum contains the following elements: discussion and role playing exercises to integrate basic knowledge of HIV/AIDS, awareness of sociocultural HIV/AIDS issues in the age of technology, and communication methods for training peer leaders in effective, interactive social media-based HIV prevention. Ethical issues related to Facebook and health interventions are integrated throughout the sessions. Training outcomes have been developed for long-term assessment of retention and efficacy. This is the first C-POL curriculum that has been adapted for use on social networking websites. Although this curriculum has been used to target African-American and Latino MSM, it has been created to allow generalization to other high-risk groups.

  13. Differential sensitivity of light-harnessing photosynthetic events in wheat and sunflower to exogenously applied ionic and nanoparticulate silver.

    Science.gov (United States)

    Pardha-Saradhi, P; Shabnam, Nisha; Sharmila, P; Ganguli, Ashok K; Kim, Hyunook

    2018-03-01

    Potential impacts of inevitable leaks of silver nanoparticles (AgNPs) into environment on human beings need attention. Owing to the vitality of photosynthesis in maintaining life and ecosystem functioning, impacts of exogenously applied nanoparticulate and Ag + on photosystem (PS)II function, which governs overall photosynthesis, in wheat and sunflower were evaluated. PSII efficiency and related Chl a fluorescence kinetics of these two plants remained unaffected by AgNPs. However, Ag + caused a significant decline in the PSII activity and related fluorescence steps in wheat, but not in sunflower. Electron flow between Q A and PQ pool was found most sensitive to Ag + . Number of active reaction centers, electron transport, trapping of absorbed light for photochemistry, and performance index declined, while dissipation of absorbed light energy as heat significantly increased in wheat exposed to Ag + . Total antioxidant activity in sunflower was least affected by both Ag and AgNPs. In contrast, in the case of wheat, the antioxidant activity was declined by Ag + but not by AgNPs. Further, the amount of silver absorbed by plants exposed to Ag + was higher than that absorbed by plants exposed to AgNPs. While wheat retained majority of Ag in its roots, sunflower showed major Ag accumulation in stem. Photosynthetic events in sunflower, unlike wheat, were least affected as no detectable Ag levels was recorded in their leaves. Our findings revealed that AgNPs seemed non/less-toxic to light harnessing photosynthetic machinery of wheat, compared to Ag + . Photosynthetic events in sunflower were not affected by Ag + , either, as its translocation to leaves was restricted. Copyright © 2017. Published by Elsevier Ltd.

  14. PbS/CdS-sensitized mesoscopic SnO2 solar cells for enhanced infrared light harnessing.

    Science.gov (United States)

    Hossain, Md Anower; Koh, Zhen Yu; Wang, Qing

    2012-05-28

    Metal oxide semiconductors with lower lying conduction band minimum and superior electron mobility are essential for efficient charge separation and collection in PbS-sensitized solar cells. In the present study, mesoscopic SnO(2) was investigated as an alternative photoanode to the commonly used TiO(2) and examined comprehensively in PbS-sensitized liquid junction solar cells. To exploit the capability of PbS in an optimized structure, cascaded nPbS/nCdS and alternate n(PbS/CdS) layers deposited by a successive ionic layer adsorption and reaction method were systematically scrutinized. It was observed that the surface of SnO(2) has greater affinity to the growth of PbS compared with TiO(2), giving rise to much enhanced light absorption. In addition, the deposition of a CdS buffer layer and a ZnS passivation layer before and after a PbS layer was found to be beneficial for efficient charge separation. Under optimized conditions, cascaded PbS/CdS-sensitized SnO(2) exhibited an unprecedented photocurrent density of 17.38 mA cm(-2) with pronounced infrared light harvesting extending beyond 1100 nm, and a power conversion efficiency of 2.23% under AM 1.5, 1 sun illumination. In comparison, TiO(2) cells fabricated under similar conditions showed much inferior performance owing to the less efficient light harnessing of long wavelength photons. We anticipate that the systematic study of PbS-sensitized solar cells utilizing different metal oxide semiconductors as electron transporters would provide useful insights and promote the development of semiconductor-sensitized mesoscopic solar cells employing panchromatic sensitizers.

  15. Treatment of Displaced Midshaft Clavicle Fractures: Figure-of-Eight Harness Versus Anterior Plate Osteosynthesis: A Randomized Controlled Trial.

    Science.gov (United States)

    Tamaoki, Marcel Jun Sugawara; Matsunaga, Fabio Teruo; Costa, Adelmo Rezende Ferreira da; Netto, Nicola Archetti; Matsumoto, Marcelo Hide; Belloti, Joao Carlos

    2017-07-19

    Most midshaft clavicle fractures affect the economically active population, which is negatively impacted by transient limb impairment during the treatment. There is still debate about the advantages and disadvantages of surgical treatment for these fractures. In this prospective randomized controlled trial, 117 patients were allocated to 1 of 2 groups: nonsurgical treatment with a figure-of-eight harness or surgical treatment with anteroinferior plate osteosynthesis. The primary outcome was upper-limb limitation measured with the Disabilities of the Arm, Shoulder and Hand (DASH) questionnaire at 6 months. Other outcomes included pain, radiographic findings, satisfaction with the cosmetic result, complications, and time to return to previous work and activities. Participants were assessed at 6 weeks, 6 months, and 1 year after the intervention. No difference between the 2 groups was detected in the DASH score at any time point (p = 0.398, 0.403, and 0.877 at 6 weeks, 6 months, and 1 year, respectively), pain levels measured with a visual analogue scale (VAS), time to return to previous activities, or dissatisfaction with the cosmetic result. Seven patients (14.9%) developed nonunion after nonsurgical treatment, a nonunion rate that was significantly higher than that in the surgical group, in which all fractures had healed (p = 0.004). The patients in the nonsurgical group had radiographic evidence of greater clavicle shortening (p study did not demonstrate a difference in limb function between patients who underwent surgical treatment and those nonsurgically treated for a dislocated midshaft clavicle fracture. Meanwhile, surgical treatment decreased the likelihood of nonunion. Therapeutic Level I. See Instructions for Authors for a complete description of levels of evidence.

  16. Har Gobind Khorana

    Indian Academy of Sciences (India)

    IAS Admin

    research in a German-speaking country. Much of the chemical literature that he had come across in his PhD work was in German and he was determined to be proficient in the language. So he went to the Swiss Federal Institute of Technology in Zurich to seek a position as a postdoctoral researcher. According to Khorana,.

  17. 5. Sexual Har

    African Journals Online (AJOL)

    bribery and coercion . This study used the definition of sexual harassment from .... envelope. Ethical consideration. The study was conducted after obtaining permission from UNZA administration. Approval was obtained from Research Ethics Committee at the University of Zambia prior to the commencement of the research.

  18. Harnessing natural ventilation benefits.

    Science.gov (United States)

    O'Leary, John

    2013-04-01

    Making sure that a healthcare establishment has a good supply of clean fresh air is an important factor in keeping patients, staff, and visitors, free from the negative effects of CO2 and other contaminants. John O'Leary of Trend Controls, a major international supplier of building energy management solutions (BEMS), examines the growing use of natural ventilation, and the health, energy-saving, and financial benefits, that it offers.

  19. Har du et problem?

    DEFF Research Database (Denmark)

    Knudsen, Sanne

    2009-01-01

    De fleste universitetsstuderende vil i større eller mindre grad opleve at skulle arbejde problemorienteret ellermed andre former for selvstændig konstruktion af vidensproblemer, om ikke andet så når de skal skrive speciale. På trods af at den problemorienterede tilgang kan forstås og udspilles på...

  20. Harnessing the atom

    International Nuclear Information System (INIS)

    1999-01-01

    Splitting the atom has had a major impact on the history of the latter part of the 20th century. This film depicts the many benefits - and also drawbacks - of nuclear technology, and describes how the International Atomic Energy Agency performs its various tasks. It touches on challenges such as the choice between major energy sources, growing concerns about the global climate, and prospects for nuclear arms control and disarmament

  1. Har vi en aftale?

    DEFF Research Database (Denmark)

    Knudsen, Hanne

    Skole og familie skal samarbejde mere og bedre. Det er alle enige om. Alligevel går det ikke altid sådan. Forældre bliver ofte skuffede over det, der opleves som skolens manglende lydhørhed.  Og lærere ser forældresamarbejdet som præget af forældrenes manglende engagement, egocentriske fokus på...

  2. Enhanced gene expression from retroviral vectors

    Directory of Open Access Journals (Sweden)

    Micklem David R

    2008-02-01

    Full Text Available Abstract Background Retroviruses are widely used to transfer genes to mammalian cells efficiently and stably. However, genetic elements required for high-level gene expression are incompatible with standard systems. The retroviral RNA genome is produced by cellular transcription and post-transcriptional processing within packaging cells: Introns present in the retroviral genomic transcript are removed by splicing, while polyadenylation signals lead to the production of ineffective truncated genomes. Furthermore strong enhancer/promoters within the retroviral payload lead to detrimental competition with the retroviral enhancer/promoter. Results By exploiting a new method of producing the retroviral genome in vitro it is possible to produce infectious retroviral particles carrying a high-level expression cassette that completely prohibits production of infectious retroviral particles by conventional methods. We produced an expression cassette comprising a strong enhancer/promoter, an optimised intron, the GFP open reading frame and a strong polyadenylation signal. This cassette was cloned into both a conventional MMLV retroviral vector and a vector designed to allow in vitro transcription of the retroviral genome by T7 RNA polymerase. When the conventional retroviral vector was transfected into packaging cells, the expression cassette drove strong GFP expression, but no infectious retrovirus was produced. Introduction of the in vitro produced uncapped retroviral genomic transcript into the packaging cells did not lead to any detectable GFP expression. However, infectious retrovirus was easily recovered, and when used to infect target primary human cells led to very high GFP expression – up to 3.5 times greater than conventional retroviral LTR-driven expression. Conclusion Retroviral vectors carrying an optimized high-level expression cassette do not produce infectious virions when introduced into packaging cells by transfection of DNA

  3. MicroRNA expression profiling to identify and validate reference genes for relative quantification in colorectal cancer

    LENUS (Irish Health Repository)

    Chang, Kah Hoong

    2010-04-29

    Abstract Background Advances in high-throughput technologies and bioinformatics have transformed gene expression profiling methodologies. The results of microarray experiments are often validated using reverse transcription quantitative PCR (RT-qPCR), which is the most sensitive and reproducible method to quantify gene expression. Appropriate normalisation of RT-qPCR data using stably expressed reference genes is critical to ensure accurate and reliable results. Mi(cro)RNA expression profiles have been shown to be more accurate in disease classification than mRNA expression profiles. However, few reports detailed a robust identification and validation strategy for suitable reference genes for normalisation in miRNA RT-qPCR studies. Methods We adopt and report a systematic approach to identify the most stable reference genes for miRNA expression studies by RT-qPCR in colorectal cancer (CRC). High-throughput miRNA profiling was performed on ten pairs of CRC and normal tissues. By using the mean expression value of all expressed miRNAs, we identified the most stable candidate reference genes for subsequent validation. As such the stability of a panel of miRNAs was examined on 35 tumour and 39 normal tissues. The effects of normalisers on the relative quantity of established oncogenic (miR-21 and miR-31) and tumour suppressor (miR-143 and miR-145) target miRNAs were assessed. Results In the array experiment, miR-26a, miR-345, miR-425 and miR-454 were identified as having expression profiles closest to the global mean. From a panel of six miRNAs (let-7a, miR-16, miR-26a, miR-345, miR-425 and miR-454) and two small nucleolar RNA genes (RNU48 and Z30), miR-16 and miR-345 were identified as the most stably expressed reference genes. The combined use of miR-16 and miR-345 to normalise expression data enabled detection of a significant dysregulation of all four target miRNAs between tumour and normal colorectal tissue. Conclusions Our study demonstrates that the top six most

  4. MicroRNA expression profiling to identify and validate reference genes for relative quantification in colorectal cancer.

    LENUS (Irish Health Repository)

    Chang, Kah Hoong

    2010-01-01

    BACKGROUND: Advances in high-throughput technologies and bioinformatics have transformed gene expression profiling methodologies. The results of microarray experiments are often validated using reverse transcription quantitative PCR (RT-qPCR), which is the most sensitive and reproducible method to quantify gene expression. Appropriate normalisation of RT-qPCR data using stably expressed reference genes is critical to ensure accurate and reliable results. Mi(cro)RNA expression profiles have been shown to be more accurate in disease classification than mRNA expression profiles. However, few reports detailed a robust identification and validation strategy for suitable reference genes for normalisation in miRNA RT-qPCR studies. METHODS: We adopt and report a systematic approach to identify the most stable reference genes for miRNA expression studies by RT-qPCR in colorectal cancer (CRC). High-throughput miRNA profiling was performed on ten pairs of CRC and normal tissues. By using the mean expression value of all expressed miRNAs, we identified the most stable candidate reference genes for subsequent validation. As such the stability of a panel of miRNAs was examined on 35 tumour and 39 normal tissues. The effects of normalisers on the relative quantity of established oncogenic (miR-21 and miR-31) and tumour suppressor (miR-143 and miR-145) target miRNAs were assessed. RESULTS: In the array experiment, miR-26a, miR-345, miR-425 and miR-454 were identified as having expression profiles closest to the global mean. From a panel of six miRNAs (let-7a, miR-16, miR-26a, miR-345, miR-425 and miR-454) and two small nucleolar RNA genes (RNU48 and Z30), miR-16 and miR-345 were identified as the most stably expressed reference genes. The combined use of miR-16 and miR-345 to normalise expression data enabled detection of a significant dysregulation of all four target miRNAs between tumour and normal colorectal tissue. CONCLUSIONS: Our study demonstrates that the top six most

  5. Intracytoplasmic stable expression of IgG1 antibody targeting NS3 helicase inhibits replication of highly efficient hepatitis C Virus 2a clone

    Directory of Open Access Journals (Sweden)

    Clementi Massimo

    2010-06-01

    Full Text Available Abstract Background Hepatitis C virus (HCV infection is a major public health problem with more than 170 million cases of chronic infections worldwide. There is no protective vaccine currently available for HCV, therefore the development of novel strategy to prevent chronic infection is important. We reported earlier that a recombinant human antibody clone blocks viral NS3 helicase activity and inhibits replication of HCV 1b virus. This study was performed further to explore the mechanism of action of this recombinant antibody and to determine whether or not this antibody inhibits replication and infectivity of a highly efficient JFH1 HCV 2a virus clone. Results The antiviral effect of intracellular expressed antibody against the HCV 2a virus strain was examined using a full-length green fluorescence protein (GFP labeled infectious cell culture system. For this purpose, a Huh-7.5 cell line stably expressing the NS3 helicase gene specific IgG1 antibody was prepared. Replication of full-length HCV-GFP chimera RNA and negative-strand RNA was strongly inhibited in Huh-7.5 cells stably expressing NS3 antibody but not in the cells expressing an unrelated control antibody. Huh-7.5 cells stably expressing NS3 helicase antibody effectively suppressed infectious virus production after natural infection and the level of HCV in the cell free supernatant remained undetectable after first passage. In contrast, Huh-7.5 cells stably expressing an control antibody against influenza virus had no effect on virus production and high-levels of infectious HCV were detected in culture supernatants over four rounds of infectivity assay. A recombinant adenovirus based expression system was used to demonstrate that Huh-7.5 replicon cell line expressing the intracellular antibody strongly inhibited the replication of HCV-GFP RNA. Conclusion Recombinant human anti-HCV NS3 antibody clone inhibits replication of HCV 2a virus and infectious virus production. Intracellular

  6. Harnessing a methane-fueled, sediment-free mixed microbial community for utilization of distributed sources of natural gas.

    Science.gov (United States)

    Marlow, Jeffrey J; Kumar, Amit; Enalls, Brandon C; Reynard, Linda M; Tuross, Noreen; Stephanopoulos, Gregory; Girguis, Peter

    2018-06-01

    Harnessing the metabolic potential of uncultured microbial communities is a compelling opportunity for the biotechnology industry, an approach that would vastly expand the portfolio of usable feedstocks. Methane is particularly promising because it is abundant and energy-rich, yet the most efficient methane-activating metabolic pathways involve mixed communities of anaerobic methanotrophic archaea and sulfate reducing bacteria. These communities oxidize methane at high catabolic efficiency and produce chemically reduced by-products at a comparable rate and in near-stoichiometric proportion to methane consumption. These reduced compounds can be used for feedstock and downstream chemical production, and at the production rates observed in situ they are an appealing, cost-effective prospect. Notably, the microbial constituents responsible for this bioconversion are most prominent in select deep-sea sediments, and while they can be kept active at surface pressures, they have not yet been cultured in the lab. In an industrial capacity, deep-sea sediments could be periodically recovered and replenished, but the associated technical challenges and substantial costs make this an untenable approach for full-scale operations. In this study, we present a novel method for incorporating methanotrophic communities into bioindustrial processes through abstraction onto low mass, easily transportable carbon cloth artificial substrates. Using Gulf of Mexico methane seep sediment as inoculum, optimal physicochemical parameters were established for methane-oxidizing, sulfide-generating mesocosm incubations. Metabolic activity required >∼40% seawater salinity, peaking at 100% salinity and 35 °C. Microbial communities were successfully transferred to a carbon cloth substrate, and rates of methane-dependent sulfide production increased more than threefold per unit volume. Phylogenetic analyses indicated that carbon cloth-based communities were substantially streamlined and were

  7. [Construction of recombinant human nerve growth factor (rh-β-NGF) eukaryotic vector and its expression in HEK293 cells].

    Science.gov (United States)

    Li, Jingchuan; Xue, Bofu; Yuan, Yuan; Ma, Mo; Zhu, Lin; Milburn, Rebecca; Le, Li; Hu, Peizhen; Ye, Jing

    2015-03-01

    Human nerve growth factor (NGF) is a nerve cell growth regulation factor, which can provide nutrition for the neurons and promote the neurites outgrowth. In order to produce large-scale recombinant human nerve growth factor (rh-beta-NGF), we constructed a plasmid vector, which can stably express the rh-beta-NGF in the HEK293 cell lines. First, the plasmid of pCMV-beta-NGF-IRES-dhfr was constructed and transformed into HEK293 cells. Then MTX pressurized filter and limiting dilution methods were used to obtain monoclonal HEK293 cell lines. After stepwise reducing serum in culture media, the cells eventually adapted to serum-free medium and secreted rh-beta-NGF. SDS-PAGE analysis revealed that the expression product owned a molecular weight of about 13 kDa and a purity of more than 50%. The peptide mapping sequencing analysis demonstrated the sequences of rh-beta-NGF matched with the theoretical ones. Later we purified this protein by ion exchange and molecular sieve chromatograph. Finally, our experimental results exhibited that the recombinant cell lines can stably express rh-beta-NGF with a high efficiency of more than 20 pg/cell x day. In addition, this protein could successfully induce differentiation of PC12 cells. In summary, our recombinant HEK293 cells can express bio-active rh-beta-NGF with great efficiency and stability, which supply a valid basis to large-scale production of rh-beta-NGF.

  8. Ectopic expression of the erythrocyte band 3 anion exchange protein, using a new avian retrovirus vector

    DEFF Research Database (Denmark)

    Fuerstenberg, S; Beug, H; Introna, M

    1990-01-01

    -erbB sequences following the splice acceptor were replaced by a cloning linker allowing insertion of foreign genes. The vector has been tested in conjunction with several helper viruses for the transmission of G418 resistance, titer, stability, transcription, and the transduction and expression of foreign genes...... in both chicken embryo fibroblasts and the QT6 quail cell line. The results show that the vector is capable of producing high titers of Neor virus from stably integrated proviruses. These proviruses express a balanced ratio of genome length to spliced transcripts which are efficiently translated...... into protein. Using the Escherichia coli beta-galactosidase gene cloned into the vector as a test construct, expression of enzyme activity could be detected in 90 to 95% of transfected target cells and in 80 to 85% of subsequently infected cells. In addition, a cDNA encoding the avian erythrocyte band 3 anion...

  9. Salicylic acid inhibits UV- and Cis-Pt-induced human immunodeficiency virus expression

    International Nuclear Information System (INIS)

    Woloschak, G.E.; Panozzo, J.; Libertin, C.R.; Schreck, S.; South Carolina Univ., Columbia, SC

    1994-01-01

    Previous studies have shown that exposure of HeLa cells stably transfected with a human immunodeficiency virus-long terminal repeat-chloramphenicol acetyl transferase (HIV-LTR-CAT) construct to UV light-induced expression from the HIV LTR. By culturing the cells with salicylic acid we demonstrated dose-dependent repression of this induced HIV expression. Repression was evident if salicylic acid was administered 2 h before, at the same time as, or up to 6 h after exposure to the DNA-damaging agent. The kinetics were similar for UV- and for cis-Pt-induced HIV expression, and induction was dependent on the UV dose or cis-Pt concentration added to the culture. These results suggest a role for the prostaglandins or the cyclooxygenase pathway or both in HIV induction mediated by DNA-damaging agents

  10. HLA reduces KIR expression level and frequency in a humanized mouse model1

    Science.gov (United States)

    van Bergen, Jeroen; Thompson, Allan; Retière, Christelle; van Pel, Melissa; Salvatori, Daniela; Lemonnier, François; Raulet, David; Trowsdale, John; Koning, Frits

    2014-01-01

    Many human Natural Killer (NK) cells are prevented from killing autologous cells by virtue of inhibitory Killer cell Immunoglobulin-like Receptors (KIR) binding `self' HLA class I molecules. Individual NK cells stably express a selected set of KIR, but it is currently disputed whether the fraction of NK cells expressing a particular inhibitory KIR is influenced by the presence of the corresponding HLA ligand. This issue has been particularly hard to tackle in a statistically meaningful way due to the extreme polymorphism of the KIR and HLA loci, with widely varying affinities for individual KIR and HLA allele combinations. Here, we use a transgenic mouse model to investigate the effect of HLA on KIR repertoire and function. In this model system, a functional interaction between HLA-Cw3 and KIR2DL2 reduced both the surface expression of KIR2DL2 as well as the frequency of KIR2DL2+ cells. PMID:23390293

  11. Transcription activator-like effector hybrids for conditional control and rewiring of chromosomal transgene expression.

    Science.gov (United States)

    Li, Yi; Moore, Richard; Guinn, Michael; Bleris, Leonidas

    2012-01-01

    The ability to conditionally rewire pathways in human cells holds great therapeutic potential. Transcription activator-like effectors (TALEs) are a class of naturally occurring specific DNA binding proteins that can be used to introduce targeted genome modifications or control gene expression. Here we present TALE hybrids engineered to respond to endogenous signals and capable of controlling transgenes by applying a predetermined and tunable action at the single-cell level. Specifically, we first demonstrate that combinations of TALEs can be used to modulate the expression of stably integrated genes in kidney cells. We then introduce a general purpose two-hybrid approach that can be customized to regulate the function of any TALE either using effector molecules or a heterodimerization reaction. Finally, we demonstrate the successful interface of TALEs to specific endogenous signals, namely hypoxia signaling and microRNAs, essentially closing the loop between cellular information and chromosomal transgene expression.

  12. Expression of the fetal hematopoiesis regulator FEV indicates leukemias of prenatal origin.

    Science.gov (United States)

    Liu, T-H; Tang, Y-J; Huang, Y; Wang, L; Guo, X-L; Mi, J-Q; Liu, L-G; Zhu, H; Zhang, Y; Chen, L; Liu, X; Zhang, L-H; Ye, Q-J; Li, B-S; Tang, J-Y; Ford, A; Enver, T; Liu, F; Chen, G-Q; Hong, D-L

    2017-05-01

    The origin of cancers is associated with etiology as well as therapeutics. Several studies reveal that malignancies in children can originate in utero. However, a diagnostic approach to distinguish between cancers initiated pre- or postnatally is absent. Here we identified a transcriptional factor FEV (fifth Ewing variant) that was expressed in fetal hematopoietic cells and became silent after birth. We characterized that FEV was essential for the self-renewal of hematopoietic stem cells (HSCs). We next found that FEV was expressed in most infant leukemia samples, but seldom in adult samples, in accord with the known prenatal origins of the former. We further determined the majority of pediatric acute lymphoid leukemia (ALL) and acute myeloid leukemia (AML) were FEV positive. Moreover, FEV knockdown markedly impaired the leukemia-propagating ability of leukemic stem cells. We therefore identified FEV is unique to fetal HSCs and stably expressed in leukemic cells of prenatal origin. It may also provide a tractable therapeutic target.

  13. Genome-Wide Constitutively Expressed Gene Analysis and New Reference Gene Selection Based on Transcriptome Data: A Case Study from Poplar/Canker Disease Interaction

    Directory of Open Access Journals (Sweden)

    Jiaping Zhao

    2017-10-01

    Full Text Available A number of transcriptome datasets for differential expression (DE genes have been widely used for understanding organismal biology, but these datasets also contain untapped information that can be used to develop more precise analytical tools. With the use of transcriptome data generated from poplar/canker disease interaction system, we describe a methodology to identify candidate reference genes from high-throughput sequencing data. This methodology will improve the accuracy of RT-qPCR and will lead to better standards for the normalization of expression data. Expression stability analysis from xylem and phloem of Populus bejingensis inoculated with the fungal canker pathogen Botryosphaeria dothidea revealed that 729 poplar transcripts (1.11% were stably expressed, at a threshold level of coefficient of variance (CV of FPKM < 20% and maximum fold change (MFC of FPKM < 2.0. Expression stability and bioinformatics analysis suggested that commonly used house-keeping (HK genes were not the most appropriate internal controls: 70 of the 72 commonly used HK genes were not stably expressed, 45 of the 72 produced multiple isoform transcripts, and some of their reported primers produced unspecific amplicons in PCR amplification. RT-qPCR analysis to compare and evaluate the expression stability of 10 commonly used poplar HK genes and 20 of the 729 newly-identified stably expressed transcripts showed that some of the newly-identified genes (such as SSU_S8e, LSU_L5e, and 20S_PSU had higher stability ranking than most of commonly used HK genes. Based on these results, we recommend a pipeline for deriving reference genes from transcriptome data. An appropriate candidate gene should have a unique transcript, constitutive expression, CV value of expression < 20% (or possibly 30% and MFC value of expression <2, and an expression level of 50–1,000 units. Lastly, when four of the newly identified HK genes were used in the normalization of expression data for 20

  14. Reliable reference genes for normalization of gene expression data in tea plants (Camellia sinensis exposed to metal stresses.

    Directory of Open Access Journals (Sweden)

    Ming-Le Wang

    Full Text Available Tea plants [Camellia sinensis (L. O. Kuntze] are an important leaf-type crop that are widely used for the production of non-alcoholic beverages in the world. Exposure to excessive amounts of heavy metals adversely affects the quality and yield of tea leaves. To analyze the molecular responses of tea plants to heavy metals, a reliable quantification of gene expression is important and of major importance herein is the normalization of the measured expression levels for the target genes. Ideally, stably expressed reference genes should be evaluated in all experimental systems. In this study, 12 candidate reference genes (i.e., 18S rRNA, Actin, CYP, EF-1α, eIF-4α, GAPDH, MON1, PP2AA3, TBP, TIP41, TUA, and UBC were cloned from tea plants, and the stability of their expression was examined systematically in 60 samples exposed to diverse heavy metals (i.e., manganese, aluminum, copper, iron, and zinc. Three Excel-based algorithms (geNorm, NormFinder, and BestKeeper were used to evaluate the expression stability of these genes. PP2AA3 and 18S rRNA were the most stably expressed genes, even though their expression profiles exhibited some variability. Moreover, commonly used reference genes (i.e., GAPDH and TBP were the least appropriate reference genes for most samples. To further validate the suitability of the analyzed reference genes, the expression level of a phytochelatin synthase gene (i.e., CsPCS1 was determined using the putative reference genes for data normalizations. Our results may be beneficial for future studies involving the quantification of relative gene expression levels in tea plants.

  15. Gene expression

    International Nuclear Information System (INIS)

    Hildebrand, C.E.; Crawford, B.D.; Walters, R.A.; Enger, M.D.

    1983-01-01

    We prepared probes for isolating functional pieces of the metallothionein locus. The probes enabled a variety of experiments, eventually revealing two mechanisms for metallothionein gene expression, the order of the DNA coding units at the locus, and the location of the gene site in its chromosome. Once the switch regulating metallothionein synthesis was located, it could be joined by recombinant DNA methods to other, unrelated genes, then reintroduced into cells by gene-transfer techniques. The expression of these recombinant genes could then be induced by exposing the cells to Zn 2+ or Cd 2+ . We would thus take advantage of the clearly defined switching properties of the metallothionein gene to manipulate the expression of other, perhaps normally constitutive, genes. Already, despite an incomplete understanding of how the regulatory switch of the metallothionein locus operates, such experiments have been performed successfully

  16. som Ungarn har fremsendt giver anledning til kommentarer eller ændringer i den risikovurdering som Danmark har gennemført i forbindelse med tilladelsen til markedsføring af majslinien MON810. " Zea mays (MON810), Indsigelse fra Ungarn vedr. nye oplysninger der påvirker risikovurderingen. Modtaget 20

    DEFF Research Database (Denmark)

    Kjellsson, Gøsta; Strandberg, Morten Tune; Damgaard, Christian

    2005-01-01

    "DMU vurderer at det præsenterede undersøgelsesmateriale fra Ungarn ikke giver nogen videnskabelig velbegrundet formodning om at den ovennævnte MON810-majs udgør en risiko for miljøet. Ungarn har således for nærværende ingen videnskabeligt baseret grund til at forbyde dyrkning og salg af såsæd in...

  17. Evaluation of Nicotiana tabacum plants transformed for the expression of verocytotoxic Escherichia coli antigens.

    Directory of Open Access Journals (Sweden)

    Angela Lombardi

    2015-07-01

    Full Text Available Two transgenic Nicotiana tabacum plants, carrying respectively the F18 adhesive fimbriae and the B subunit of verocytotoxin genes from O138 Verocytotoxic E.coli serotype were developed as a model of edible vaccine. Tobacco plants were transformed by agroinfection according to Rossi et al. (2013 stably.  The F18 adhesive fimbriae and VT2e B-subunit were placed under control of the GLOB promoter for the seed-specific protein expression. Agrobacterium tumefaciens binary vector system is an efficient tool to transform plant cells; however, the exogenous gene integrates at semi-random into the nuclear chromosome. PCR products, using specific oligonucleotides putatively encoding the B-subunit of VT2e-B and F18 fimbriae were identified on agarose gel (1.5% - 0.9% as bands with a length of 270 and 519 base pairs, respectively. We showed that the foreign VT2e-B and F18 fimbriae genes were stably integrated into the tobacco genome. Northern blot and Western blot analyses carried out respectively on total mRNA and total soluble protein extract obtained from seeds. For each line, the obtained amount of antigens is sufficient for subsequent oral immunization trials. Three lines of tobacco seeds (F18, VT2e-B, and WT were seeded in homogeneous conditions and were harvested simultaneously. Tobacco plants were analysed also by optical and electronic microscope in different phases of growth. Germination of transgenic seeds were delayed of three/five days compared to WT in two replicated experiments, suggesting that genetic manipulation may influenced mechanisms leading to germination. In conclusion the genes coding for VT2e-B and the F18 are stably maintained in the seeds and obtained tobacco seeds represent a valid strategy to ferry antigenic proteins to the gut and a promising non-invasive method of vaccination in pig industry.

  18. Transcriptional expression of type I interferon response genes and stability of housekeeping genes in the human endometrium and endometriosis

    DEFF Research Database (Denmark)

    Vestergaard, Anna L; Knudsen, Ulla B; Munk, Torben

    2011-01-01

    Endometriosis is a painful chronic female disease defined by the presence of endometrial tissue implants in ectopic locations. The pathogenesis is much debated, and type I interferons could be involved. The expression of genes of the type I interferon response were profiled by a specific PCR Array...... of RNA obtained from ectopic and eutopic endometrium collected from 9 endometriosis patients and 9 healthy control women. Transcriptional expression levels of selected interferon-regulated and housekeeping genes were investigated by real-time quantitative reverse transcriptase PCR (qRT-PCR). Stably...... expressed housekeeping genes for valid normalization of transcriptional studies of endometrium and endometriosis have not yet been published. Here, seven housekeeping genes were evaluated for stability using the GeNorm and NormFinder software. A normalization factor based on HMBS, TBP, and YWHAZ expression...

  19. Algevir: An Expression System for Microalgae Based on Viral Vectors

    Science.gov (United States)

    Bañuelos-Hernández, Bernardo; Monreal-Escalante, Elizabeth; González-Ortega, Omar; Angulo, Carlos; Rosales-Mendoza, Sergio

    2017-01-01

    The use of recombinant algae for the production of valuable compounds is opening promising biotechnological applications. However, the development of efficient expression approaches is still needed to expand the exploitation of microalgae in biotechnology. Herein, the concept of using viral expression vectors in microalgae was explored for the first time. An inducible geminiviral vector leading to Rep-mediated replication of the expression cassette allowed the production of antigenic proteins at high levels. This system, called Algevir, allows the production of complex viral proteins (GP1 from Zaire ebolavirus) and bacterial toxin subunits (B subunit of the heat-labile Escherichia coli enterotoxin), which retained their antigenic activity. The highest achieved yield was 1.25 mg/g fresh biomass (6 mg/L of culture), which was attained 3 days after transformation. The Algevir system allows for a fast and efficient production of recombinant proteins, overcoming the difficulties imposed by the low yields and unstable expression patterns frequently observed in stably transformed microalgae at the nuclear level; as well as the toxicity of some target proteins. PMID:28713333

  20. Algevir: An Expression System for Microalgae Based on Viral Vectors

    Directory of Open Access Journals (Sweden)

    Bernardo Bañuelos-Hernández

    2017-06-01

    Full Text Available The use of recombinant algae for the production of valuable compounds is opening promising biotechnological applications. However, the development of efficient expression approaches is still needed to expand the exploitation of microalgae in biotechnology. Herein, the concept of using viral expression vectors in microalgae was explored for the first time. An inducible geminiviral vector leading to Rep-mediated replication of the expression cassette allowed the production of antigenic proteins at high levels. This system, called Algevir, allows the production of complex viral proteins (GP1 from Zaire ebolavirus and bacterial toxin subunits (B subunit of the heat-labile Escherichia coli enterotoxin, which retained their antigenic activity. The highest achieved yield was 1.25 mg/g fresh biomass (6 mg/L of culture, which was attained 3 days after transformation. The Algevir system allows for a fast and efficient production of recombinant proteins, overcoming the difficulties imposed by the low yields and unstable expression patterns frequently observed in stably transformed microalgae at the nuclear level; as well as the toxicity of some target proteins.

  1. Cellular expression of gH confers resistance to herpes simplex virus type-1 entry

    International Nuclear Information System (INIS)

    Scanlan, Perry M.; Tiwari, Vaibhav; Bommireddy, Susmita; Shukla, Deepak

    2003-01-01

    Entry of herpes simplex virus-1 (HSV-1) into cells requires a concerted action of four viral glycoproteins gB, gD, and gH-gL. Previously, cell surface expression of gD had been shown to confer resistance to HSV-1 entry. To investigate any similar effects caused by other entry glycoproteins, gB and gH-gL were coexpressed with Nectin-1 in Chinese hamster ovary (CHO) cells. Interestingly, cellular expression of gB had no effect on HSV-1(KOS) entry. In contrast, entry was significantly reduced in cells expressing gH-gL. This effect was further analyzed by expressing gH and gL separately. Cells expressing gL were normally susceptible, whereas gH-expressing cells were significantly resistant. Further experiments suggested that the gH-mediated interference phenomenon was not specific to any particular gD receptor and was also observed in gH-expressing HeLa cells. Moreover, contrary to a previous report, gL-independent cell surface expression of gH was detected in stably transfected CHO cells, possibly implicating cell surface gH in the interference phenomenon. Thus, taken together these findings indicate that cellular expression of gH interferes with HSV-1 entry

  2. Hvad har idræt og sundhedsfremme i skoleregi med demokrati at gøre? en sociologisk analyse af idrætspolitik i gymnasieskolen

    DEFF Research Database (Denmark)

    Nielsen, Stine Frydendal; Thing, Lone Friis

    Den internationale forskning påpeger at unge bevæger sig for lidt, og at det har alvorlige konsekvenser for helbredet (WHO 2004; Strong et al 2005). Specielt pigerne bliver inaktive i ungdomsårene (Coleman et al 2008; Ward et al 2006; Pate et al 2005; Allender et al 2005). Skævvridningen i...... idrætsdeltagelsen gælder også i det danske samfund (Pilgård 2009; Osler et al 2001). Internationale studier viser endvidere at fastholdelse af sundhedsinterventioner er svært, og resultaterne påpeger, at der fortsat er brug for viden om hvordan fastholdelse af sundhedsfremme interventioner med henblik på at fremme...... fysisk aktivitet kan ske (Laitakari et al 1996). En del forskningsundersøgelser har forsøgt at have skolen som en platform for sundhedsfremme – bl.a. ved at forsøge at øge idrætsaktiviteten i faget physical education (PE) (Fairclough et al 2008; Fairclough & Stratton 2005; Pate et al 2005). GISU...

  3. Solution structure of the NEAT (NEAr Transporter) domain from IsdH/HarA: the human hemoglobin receptor in Staphylococcus aureus.

    Science.gov (United States)

    Pilpa, Rosemarie M; Fadeev, Evgeny A; Villareal, Valerie A; Wong, Melissa L; Phillips, Martin; Clubb, Robert T

    2006-07-07

    During infections the pathogen Staphylococcus aureus procures the essential nutrient iron from its host using iron-regulated surface determinant (Isd) proteins, which scavenge heme bound iron from host hemoproteins. Four Isd proteins are displayed in the cell wall, where they function as receptors for host proteins and heme. Each of the receptors contains one or more copies of a recently discovered domain called NEAT (NEAr Transporter) that has been shown to mediate protein binding. Here we report the three-dimensional solution structure of the NEAT domain from the IsdH/HarA protein, which is the hemoglobin receptor in the Isd system. This is the first structure of a NEAT domain and reveals that they adopt a beta sandwich fold that consists of two five-stranded antiparallel beta sheets. Although unrelated at the primary sequence level, our results indicate that NEAT domains belong to the immunoglobulin superfamily. Binding studies indicate that two IsdH/HarA NEAT domains bind a single molecule of methemoglobin, while the distantly related NEAT domain from the S. aureus IsdC protein binds only heme. A comparison of their primary sequences in light of the new structure is used to predict the hemoglobin and heme binding surfaces on NEAT domains.

  4. Further development of the pneumatic method to harness hydropower and its experimental implementation in the State of Maine. Final report, [February 15, 1990--February 14, 1993

    Energy Technology Data Exchange (ETDEWEB)

    Gorlov, A.M.

    1994-03-01

    This report contains conclusive results of the research project entitled ``Further Development of the Pneumatic Method to Harness Hydropower and its Experimental Implementation in the State of Maine`` sponsored by the US Department of Energy (Contract DE-FG02-91ER12113). The results obtained by this research are considerably beyond the original goals anticipated by the contract which were a theoretical study of the method only and its possible applications. In fact, the success of the analytical research program has allowed us to move on to development, construction and testing of a physical model of the hydro-air power converter and, subsequently, to development of a well equipped hydro-pneumatic power laboratory at Northeastern University. Photographs la and 2a show both the laboratory and the model. Good performance of the model proves that the hydro-pneumatic concept holds much promise for development of an ecologically safe and commercially attractive novel approach to harnessing ultra low-head hydropower. As a result, private companies have started to support this new technology, and to invest money in its further development and construction of demonstration power plants (Appendix 1). Visitors at the Northeastern University laboratory often praise this new technique, as is attested by the articles in the Wall Street Journal (Appendix 2) and the Technology Review (Appendix 3).

  5. Fibroblast activation protein-α-expressing fibroblasts promote the progression of pancreatic ductal adenocarcinoma.

    Science.gov (United States)

    Kawase, Tomoya; Yasui, Yumiko; Nishina, Sohji; Hara, Yuichi; Yanatori, Izumi; Tomiyama, Yasuyuki; Nakashima, Yoshihiro; Yoshida, Koji; Kishi, Fumio; Nakamura, Masafumi; Hino, Keisuke

    2015-09-02

    Pancreatic ductal adenocarcinoma (PDAC) is characterized by an extensive desmoplastic stromal response. Fibroblast activation protein-α (FAP) is best known for its presence in stromal cancer-associated fibroblasts (CAFs). Our aim was to assess whether FAP expression was associated with the prognosis of patients with PDAC and to investigate how FAP expressing CAFs contribute to the progression of PDAC. FAP expression was immunohistochemically assessed in 48 PDAC specimens. We also generated a fibroblastic cell line stably expressing FAP, and examined the effect of FAP-expressing fibroblasts on invasiveness and the cell cycle in MiaPaCa-2 cells (a pancreatic cancer cell line). Stromal FAP expression was detected in 98% (47/48) of the specimens of PDAC, with the intensity being weak in 16, moderate in 19, and strong in 12 specimens, but was not detected in the 3 control noncancerous pancreatic specimens. Patients with moderate or strong FAP expression had significantly lower cumulative survival rates than those with negative or weak FAP expression (mean survival time; 352 vs. 497 days, P = 0.006). Multivariate analysis identified moderate to strong expression of FAP as one of the factors associated with the prognosis in patients with PDAC. The intensity of stromal FAP expression was also positively correlated to the histological differentiation of PDAC (P fibroblasts promoted the invasiveness of MiaPaCa-2 cells more intensively than fibroblasts not expressing FAP. Coculture with FAP-expressing fibroblasts significantly activated cell cycle shift in MiaPaCa-2 cells compared to coculture with fibroblasts not expressing FAP. Furthermore, coculture with FAP expressing fibroblasts inactivated retinoblastoma (Rb) protein, an inhibitor of cell cycle progression, in MiaPaCa-2 cells by promoting phosphorylation of Rb. The present in vitro results and the association of FAP expression with clinical outcomes provide us with a better understanding of the effect of FAP-expressing

  6. Reference genes for gene expression analysis by real-time reverse transcription polymerase chain reaction of renal cell carcinoma.

    Science.gov (United States)

    Bjerregaard, Henriette; Pedersen, Shona; Kristensen, Søren Risom; Marcussen, Niels

    2011-12-01

    Differentiation between malignant renal cell carcinoma and benign oncocytoma is of great importance to choose the optimal treatment. Accurate preoperative diagnosis of renal tumor is therefore crucial; however, existing imaging techniques and histologic examinations are incapable of providing an optimal differentiation profile. Analysis of gene expression of molecular markers is a new possibility but relies on appropriate standardization to compare different samples. The aim of this study was to identify stably expressed reference genes suitable for the normalization of results extracted from gene expression analysis of renal tumors. Expression levels of 8 potential reference genes (ATP5J, HMBS, HPRT1, PPIA, TBP, 18S, GAPDH, and POLR2A) were examined by real-time reverse transcription polymerase chain reaction in tumor and normal tissue from removed kidneys from 13 patients with renal cell carcinoma and 5 patients with oncocytoma. The expression levels of genes were compared by gene stability value M, average gene stability M, pairwise variation V, and coefficient of variation CV. More candidates were not suitable for the purpose, but a combination of HMBS, PPIA, ATP5J, and TBP was found to be the best combination with an average gene stability value M of 0.9 and a CV of 0.4 in the 18 tumors and normal tissues. A combination of 4 genes, HMBS, PPIA, ATP5J, and TBP, is a possible reference in renal tumor gene expression analysis by reverse transcription polymerase chain reaction. A combination of four genes, HMBS, PPIA, ATP5J and TBP, being stably expressed in tissues from RCC is possible reference genes for gene expression analysis.

  7. Identification of optimal reference genes for gene expression normalization in a wide cohort of endometrioid endometrial carcinoma tissues.

    Directory of Open Access Journals (Sweden)

    Chiara Romani

    Full Text Available Accurate normalization is a primary component of a reliable gene expression analysis based on qRT-PCR technique. While the use of one or more reference genes as internal controls is commonly accepted as the most appropriate normalization strategy, many qPCR-based published studies still contain data poorly normalized and reference genes arbitrarily chosen irrespective of the particular tissue and the specific experimental design. To date, no validated reference genes have been identified for endometrial cancer tissues. In this study, 10 normalization genes (GAPDH, B2M, ACTB, POLR2A, UBC, PPIA, HPRT1, GUSB, TBP, H3F3A belonging to different functional and abundance classes in various tissues and used in different studies, were analyzed to determine their applicability. In total, 100 endometrioid endometrial cancer samples, which were carefully balanced according to their tumor grade, and 29 normal endometrial tissues were examined using SYBR Green Real-Time RT-PCR. The expression stability of candidate reference genes was determined and compared by means of geNorm and NormFinder softwares. Both algorithms were in agreement in identifying GAPDH, H3F3A, PPIA, and HPRT1 as the most stably expressed genes, only differing in their ranking order. Analysis performed on the expression levels of all candidate genes confirm HPRT1 and PPIA as the most stably expressed in the study groups regardless of sample type, to be used alone or better in combination. As the stable expression of HPRT1 and PPIA between normal and tumor endometrial samples fulfill the basic requirement of a reference gene to be used for normalization purposes, HPRT1 expression showed significant differences between samples from low-grade and high-grade tumors. In conclusion, our results recommend the use of PPIA as a single reference gene to be considered for improved reliability of normalization in gene expression studies involving endometrial tumor samples at different tumor degrees.

  8. A rapid and versatile method for harnessing scFv antibody fragments with various biological effector functions

    NARCIS (Netherlands)

    Helfrich, W; Haisma, HJ; Magdolen, [No Value; Luther, T; Bom, VJJ; Westra, J.; van der Hoeven, R; Kroesen, BJ; Molema, G; de Leij, L

    2000-01-01

    A versatile expression vector is described for the rapid construction and evaluation of bispecific scFvs and scFv-based fusion proteins. An important feature of this vector is the presence of two multiple cloning sites (MCS) separated by an in frame linker sequence. The first MCS was specifically

  9. A new β-estradiol-inducible vector set that facilitates easy construction and efficient expression of transgenes reveals CBL3-dependent cytoplasm to tonoplast translocation of CIPK5.

    Science.gov (United States)

    Schlücking, Kathrin; Edel, Kai H; Köster, Philipp; Drerup, Maria M; Eckert, Christian; Steinhorst, Leonie; Waadt, Rainer; Batistic, Oliver; Kudla, Jörg

    2013-11-01

    Transient and stable expression of transgenes is central to many investigations in plant biology research. Chemical regulation of expression can circumvent problems of plant lethality caused by constitutive overexpression or allow inducible knock (out/down) approaches. Several chemically inducible or repressible systems have been described and successfully applied. However, cloning and application-specific modification of most available inducible expression systems have been limited and remained complicated due to restricted cloning options. Here we describe a new set of 57 vectors that enable transgene expression in transiently or stably transformed cells. All vectors harbor a synthetically optimized XVE expression cassette, allowing β-estradiol mediated protein expression. Plasmids are equipped with the reporter genes GUS, GFP, mCherry, or with HA and StrepII epitope tags and harbor an optimized multiple cloning site for flexible and simple cloning strategies. Moreover, the vector design allows simple substitution of the driving promoter to achieve tissue-specificity or to modulate expression ranges of inducible transgene expression. We report details of the kinetics and dose-dependence of expression induction in Arabidopsis leaf mesophyll protoplasts, transiently transformed Nicotiana benthamiana leaves, and stably transformed Arabidopsis plants. Using these vectors, we investigated the influence of CBL (Calcineurin B-like) protein expression on the subcellular localization of CIPKs (Calcineurin B-like interacting protein kinases). These analyses uncovered that induced co-expression of CBL3 is fully sufficient for dynamic translocation of CIPK5 from the cytoplasm to the tonoplast. Thus, the vector system presented here facilitates a broad range of research applications.

  10. CXCL12 expression promotes esophageal squamous cell carcinoma proliferation and worsens the prognosis

    International Nuclear Information System (INIS)

    Uchi, Yusuke; Takeuchi, Hiroya; Matsuda, Sachiko; Saikawa, Yoshiro; Kawakubo, Hirofumi; Wada, Norihito; Takahashi, Tsunehiro; Nakamura, Rieko; Fukuda, Kazumasa; Omori, Tai; Kitagawa, Yuko

    2016-01-01

    The chemokine CXCL12 and its corresponding receptor CXCR4 are key players in the development of several cancers. Therefore, we hypothesized that there is a functional causality between CXCL12 expression and tumor progression in patients with esophageal squamous cell carcinoma (ESCC). We performed an immunohistochemical analysis in 79 consecutive patients with ESCC. We performed in vitro and in vivo cell proliferation assays using ESCC cell lines and a newly established transfectant stably overexpressing CXCL12. Immunohistochemistry revealed positive CXCR4 and CXCL12 expression in 48 (61 %) and 62 (78 %) patients, respectively. Additionally, the expression levels did not significantly correlate with any clinicopathological factors. The MIB-1 proliferation index was markedly higher in ESCC with a positive expression of CXCR4 or CXCL12. Positive CXCL12 expression was significantly correlated with lower recurrence-free survival (RFS, p = 0.02). Cox’s hazard models revealed CXCL12 expression as an independent predictive factor for recurrence. In vitro, CXCL12 exposure or overexpression enhanced ESCC proliferation; and AMD3100, a specific inhibitor of CXCR4, equally decreased proliferation irrespective of CXCL12 exposure or overexpression. In the mouse model, AMD3100 significantly decreased ESCC tumor size (p = 0.03). CXCL12 stimulates ESCC proliferation, and its expression levels are related to lower RFS in patients with ESCC. Our findings indicate that positive CXCL12 expression may be a useful marker for predicting the outcome in patients with ESCC and is a potentially new therapeutic target for ESCC

  11. Toward an ideal animal model to trace donor cell fates after stem cell therapy: production of stably labeled multipotent mesenchymal stem cells from bone marrow of transgenic pigs harboring enhanced green fluorescence protein gene.

    Science.gov (United States)

    Hsiao, F S H; Lian, W S; Lin, S P; Lin, C J; Lin, Y S; Cheng, E C H; Liu, C W; Cheng, C C; Cheng, P H; Ding, S T; Lee, K H; Kuo, T F; Cheng, C F; Cheng, W T K; Wu, S C

    2011-11-01

    The discovery of postnatal mesenchymal stem cells (MSC) with their general multipotentiality has fueled much interest in the development of cell-based therapies. Proper identification of transplanted MSC is crucial for evaluating donor cell distribution, differentiation, and migration. Lack of an efficient marker of transplanted MSC has precluded our understanding of MSC-related regenerative studies, especially in large animal models such as pigs. In the present study, we produced transgenic pigs harboring an enhanced green fluorescent protein (EGFP) gene. The pigs provide a reliable and reproducible source for obtaining stable EGFP-labeled MSC, which is very useful for donor cell tracking after transplantation. The undifferentiated EGFP-tagged MSC expressed a greater quantity of EGFP while maintaining MSC multipotentiality. These cells exhibited homogeneous surface epitopes and possessed classic trilineage differentiation potential into osteogenic, adipogenic, and chondrogenic lineages, with robust EGFP expression maintained in all differentiated progeny. Injection of donor MSC can dramatically increase the thickness of infarcted myocardium and improve cardiac function in mice. Moreover, the MSC, with their strong EGFP expression, can be easily distinguished from the background autofluorescence in myocardial infarcts. We demonstrated an efficient, effective, and easy way to identify MSC after long-term culture and transplantation. With the transgenic model, we were able to obtain stem or progenitor cells in earlier passages compared with the transfection of traceable markers into established MSC. Because the integration site of the transgene was the same for all cells, we lessened the potential for positional effects and the heterogeneity of the stem cells. The EGFP-transgenic pigs may serve as useful biomedical and agricultural models of somatic stem cell biology.

  12. Heat radiation approach for harnessing heat of the cook stove to generate electricity for lighting system and charging of mobile phone

    Science.gov (United States)

    Muñoz, Rodrigo C., Jr.; Manansala, Chad Deo G.

    2018-01-01

    This study is based on the potential of thermoelectric coupling such as the thermoelectric cooler module. A thermoelectric cooler converts the heat coming from the cook stove into electricity and store in a battery. A dc-dc boost converter will be used to produce enough voltage to light a minimum house dwelling or charge phone battery. This device will be helpful to those that faces a problem on electricity especially in the isolated areas. The study aims (1) to harness heat from the cook stove up to 110 °C (2) To automatically cool-off the system to protect the thermoelectric cooler from damage due to excessive heat using an electronic solenoid; (3) To store energy harnessed in the battery; (4) To amplify the output voltages of the battery using DC to DC boost converter for lighting system and charging of mobile phone battery. From various tests conducted, it can fully charge a mobile phone in 3 hours observing the unit’s battery voltage drop from 4.06V to 3.98V. In the testing it used different orientation of steel rod by conduction to transfer heat and by radiation through tubular steel with its different dimensions. Most recent testing proved that the 2x2x9 tubular steel by radiation had the best result. The temperature reached more than a hundred degree Celsius that met the objective. The test resulted of boosting the voltage of the battery output from 3.7V to 4.96V on the average. The boosted voltage decrease as the system’s cool-off mechanism operated when the temperature reached above 110 degree Celsius decreasing output voltage to 0.8V resulting the boosted voltage to drop to zero. Therefore, the proponents concluded that heat waste can be converted to electrical energy by harnessing heat through radiation, with the help of TEC that generates voltage for lighting and can be boosted to be used for mobile charging. Furthermore, the study proved that the excess heat can damaged the TEC which was prevented by using of cooling-off mechanism, making it more

  13. Validation of reference genes for quantitative RT-PCR studies of gene expression in perennial ryegrass (Lolium perenne L.

    Directory of Open Access Journals (Sweden)

    Thrush Anthony

    2010-01-01

    Full Text Available Abstract Background Perennial ryegrass (Lolium perenne L. is an important pasture and turf crop. Biotechniques such as gene expression studies are being employed to improve traits in this temperate grass. Quantitative reverse transcription-polymerase chain reaction (qRT-PCR is among the best methods available for determining changes in gene expression. Before analysis of target gene expression, it is essential to select an appropriate normalisation strategy to control for non-specific variation between samples. Reference genes that have stable expression at different biological and physiological states can be effectively used for normalisation; however, their expression stability must be validated before use. Results Existing Serial Analysis of Gene Expression data were queried to identify six moderately expressed genes that had relatively stable gene expression throughout the year. These six candidate reference genes (eukaryotic elongation factor 1 alpha, eEF1A; TAT-binding protein homolog 1, TBP-1; eukaryotic translation initiation factor 4 alpha, eIF4A; YT521-B-like protein family protein, YT521-B; histone 3, H3; ubiquitin-conjugating enzyme, E2 were validated for qRT-PCR normalisation in 442 diverse perennial ryegrass (Lolium perenne L. samples sourced from field- and laboratory-grown plants under a wide range of experimental conditions. Eukaryotic EF1A is encoded by members of a multigene family exhibiting differential expression and necessitated the expression analysis of different eEF1A encoding genes; a highly expressed eEF1A (h, a moderately, but stably expressed eEF1A (s, and combined expression of multigene eEF1A (m. NormFinder identified eEF1A (s and YT521-B as the best combination of two genes for normalisation of gene expression data in perennial ryegrass following different defoliation management in the field. Conclusions This study is unique in the magnitude of samples tested with the inclusion of numerous field-grown samples

  14. Expression, characterisation and antigenicity of a truncated Hendra virus attachment protein expressed in the protozoan host Leishmania tarentolae.

    Science.gov (United States)

    Fischer, Kerstin; dos Reis, Vinicius Pinho; Finke, Stefan; Sauerhering, Lucie; Stroh, Eileen; Karger, Axel; Maisner, Andrea; Groschup, Martin H; Diederich, Sandra; Balkema-Buschmann, Anne

    2016-02-01

    Hendra virus (HeV) is an emerging zoonotic paramyxovirus within the genus Henipavirus that has caused severe morbidity and mortality in humans and horses in Australia since 1994. HeV infection of host cells is mediated by the membrane bound attachment (G) and fusion (F) glycoproteins, that are essential for receptor binding and fusion of viral and cellular membranes. The eukaryotic unicellular parasite Leishmania tarentolae has recently been established as a powerful tool to express recombinant proteins with mammalian-like glycosylation patterns, but only few viral proteins have been expressed in this system so far. Here, we describe the purification of a truncated, Strep-tag labelled and soluble version of the HeV attachment protein (sHeV G) expressed in stably transfected L. tarentolae cells. After Strep-tag purification the identity of sHeV G was confirmed by immunoblotting and mass spectrometry. The functional binding of sHeV G to the HeV cell entry receptor ephrin-B2 was confirmed in several binding assays. Generated polyclonal rabbit antiserum against sHeV G reacted with both HeV and Nipah virus (NiV) G proteins in immunofluorescence assay and efficiently neutralised NiV infection, thus further supporting the preserved antigenicity of the purified protein. Copyright © 2015 The Authors. Published by Elsevier B.V. All rights reserved.

  15. Upregulation of Lhx8 increase VAChT expression and ACh release in neuronal cell line SHSY5Y.

    Science.gov (United States)

    Li, Haoming; Jin, Guohua; Zhu, Peipei; Zou, Linqing; Shi, Jinhong; Yi, Xin; Zhang, Xinhua; Tian, Meiling; Qin, Jianbing

    2014-01-24

    Lhx8 is a transcription factor for cholinergic differentiation. Our previous experiments found upregulation of Lhx8 promoted cholinergic neuronal differentiation of hippocampal neural stem/progenitor cells or hippocampal newborn neurons in vitro. However, the role of Lhx8 in VAChT expression and ACh release is still less understood. In this report, we transfected Lhx8 cDNA into neuronal cell line SHSY5Y by lentiviral vectors to acquire the cells which stably expressed high level of Lhx8. Using this cell model, we provided experimental evidence that increasing Lhx8 upregulated the expression of ChAT and VAChT, and also increased the ACh release in culture medium. We suggested that Lhx8 overexpression is a useful strategy to increase the release of ACh and maybe of therapeutic value to neurodegenerative diseases. Copyright © 2013 Elsevier Ireland Ltd. All rights reserved.

  16. Engelsk som tredjespråk: Har lærere kompetanse til å støtte flerspråklighet?

    Directory of Open Access Journals (Sweden)

    Anne Dahl

    2016-01-01

    Full Text Available Denne artikkelen handler om norske engelsklæreres kompetanse i å arbeide med barn som har et annet morsmål enn norsk, og som lærer engelsk som fremmedspråk. Problemstillingene er "Hva slags utdanning og kunnskap innen flerspråklighet har engelsklærere i Norge?," "I hvilken grad føler engelsklærere i Norge seg forberedt på å undervise i flerkulturelle og flerspråklige klasserom?" og "Hvilken type kunnskap, ferdigheter og ressurser føler de behov for?". Vi presenterer resultater fra en nasjonal spørreundersøkelse hvor i alt 176 engelsklærere deltok. I tillegg presenterer vi resultater fra fokusintervjuer med i alt fire lærere fra to skoler. Resultatene viser at selv om engelsklærerne til en viss grad føler seg forberedt på arbeid med elever som ikke har norsk som morsmål, har svært få av dem utdanning som fokuserer på flerspråklighet. Svarene fra spørreundersøkelsen og fra intervjuene viser at engelsklærerne gjerne vil ha mer kompetanse på dette området. Vi vil derfor understreke betydningen av utdanning i teori og praksis om flerspråklig utvikling og flerkulturell utdanningsteori, og ikke minst betydningen av tilgang til faglig utvikling som fokuserer på flerspråklighet i skolen og i samfunnet for engelsklærere i Norge.Nøkkelord: engelsklærere, kompetanse, språkpedagogisk kunnskap,flerspråklighet, lærerutdanningAbstractThis paper examines the extent to which English teachers in Norway are prepared to work with children whose mother tongue is not Norwegian, and who are acquiring English as a foreign language. The research questions are “What type of education and knowledge in the area of multilingualism do English teachers in Norway have?,” To what extent do English teachers in Norway feel they are prepared to teach in a multicultural and multilingual classroom?,” and “What kinds of knowledge, skills and resources do they feel a need for?” We present the results of a national survey completed by 176

  17. Characterization of the biomass of a hybrid anaerobic reactor (HAR with two types of support material during the treatment of the coffee wastewater

    Directory of Open Access Journals (Sweden)

    Vivian Galdino da Silva

    2013-06-01

    Full Text Available This study investigated the microbiology of a hybrid anaerobic reactor (HAR in the removal of pollutant loads. This reactor had the same physical structure of an UASB reactor, however with minifilters inside containing two types of support material: expanded clay and gravel. Two hydraulic retention times (HRT of 24h and 18h were evaluated at steady-state conditions, resulting in organic loading rates (OLR of 0.032 and 0.018 kgDBO5m-3d-1 and biological organic loading rates (BOLR of 0,0015 and 0.001 kgDBO5kgSVT- 1d¹, respectively. The decrease in concentration of organic matter in the influent resulted an endogenous state of the biomass in the reactor. The expanded clay was the best support material for biofilm attachment.

  18. Tec protein tyrosine kinase inhibits CD25 expression in human T-lymphocyte.

    Science.gov (United States)

    Susaki, Kentaro; Kitanaka, Akira; Dobashi, Hiroaki; Kubota, Yoshitsugu; Kittaka, Katsuharu; Kameda, Tomohiro; Yamaoka, Genji; Mano, Hiroyuki; Mihara, Keichiro; Ishida, Toshihiko

    2010-01-04

    The Tec protein tyrosine kinase (PTK) belongs to a group of structurally related nonreceptor PTKs that also includes Btk, Itk, Rlk, and Bmx. Previous studies have suggested that these kinases play important roles in hematopoiesis and in the lymphocyte signaling pathway. Despite evidence suggesting the involvement of Tec in the T-lymphocyte activation pathway via T-cell receptor (TCR) and CD28, Tec's role in T-lymphocytes remains unclear because of the lack of apparent defects in T-lymphocyte function in Tec-deficient mice. In this study, we investigated the role of Tec in human T-lymphocyte using the Jurkat T-lymphoid cell line stably transfected with a cDNA encoding Tec. We found that the expression of wild-type Tec inhibited the expression of CD25 induced by TCR cross-linking. Second, we observed that LFM-A13, a selective inhibitor of Tec family PTK, rescued the suppression of TCR-induced CD25 expression observed in wild-type Tec-expressing Jurkat cells. In addition, expression of kinase-deleted Tec did not alter the expression level of CD25 after TCR ligation. We conclude that Tec PTK mediates signals that negatively regulate CD25 expression induced by TCR cross-linking. This, in turn, implies that this PTK plays a role in the attenuation of IL-2 activity in human T-lymphocytes.

  19. Regulation of gene expression in ovarian cancer cells by luteinizing hormone receptor expression and activation

    International Nuclear Information System (INIS)

    Cui, Juan; Miner, Brooke M; Eldredge, Joanna B; Warrenfeltz, Susanne W; Dam, Phuongan; Xu, Ying; Puett, David

    2011-01-01

    Since a substantial percentage of ovarian cancers express gonadotropin receptors and are responsive to the relatively high concentrations of pituitary gonadotropins during the postmenopausal years, it has been suggested that receptor activation may contribute to the etiology and/or progression of the neoplasm. The goal of the present study was to develop a cell model to determine the impact of luteinizing hormone (LH) receptor (LHR) expression and LH-mediated LHR activation on gene expression and thus obtain insights into the mechanism of gonadotropin action on ovarian surface epithelial (OSE) carcinoma cells. The human ovarian cancer cell line, SKOV-3, was stably transfected to express functional LHR and incubated with LH for various periods of time (0-20 hours). Transcriptomic profiling was performed on these cells to identify LHR expression/activation-dependent changes in gene expression levels and pathways by microarray and qRT-PCR analyses. Through comparative analysis on the LHR-transfected SKOV-3 cells exposed to LH, we observed the differential expression of 1,783 genes in response to LH treatment, among which five significant families were enriched, including those of growth factors, translation regulators, transporters, G-protein coupled receptors, and ligand-dependent nuclear receptors. The most highly induced early and intermediate responses were found to occupy a network impacting transcriptional regulation, cell growth, apoptosis, and multiple signaling transductions, giving indications of LH-induced apoptosis and cell growth inhibition through the significant changes in, for example, tumor necrosis factor, Jun and many others, supportive of the observed cell growth reduction in in vitro assays. However, other observations, e.g. the substantial up-regulation of the genes encoding the endothelin-1 subtype A receptor, stromal cell-derived factor 1, and insulin-like growth factor II, all of which are potential therapeutic targets, may reflect a positive

  20. Stability and inheritance of endosperm-specific expression of two transgenes in progeny from crossing independently transformed barley plants.

    Science.gov (United States)

    Choi, Hae-Woon; Yu, Xiao-Hong; Lemaux, Peggy G; Cho, Myeong-Je

    2009-08-01

    To study stability and inheritance of two different transgenes in barley, we crossed a homozygous T(8) plant, having uidA (or gus) driven by the barley endosperm-specific B(1)-hordein promoter (localized in the near centromeric region of chromosome 7H) with a second homozygous T(4) plant, having sgfp(S65T) driven by the barley endosperm-specific D-hordein promoter (localized on the subtelomeric region of chromosome 2H). Both lines stably expressed the two transgenes in the generations prior to the cross. Three independently crossed F(1) progeny were analyzed by PCR for both uidA and sgfp(S65T) in each plant and functional expression of GUS and GFP in F(2) seeds followed a 3:1 Mendelian segregation ratio and transgenes were localized by FISH to the same location as in the parental plants. FISH was used to screen F(2) plants for homozygosity of both transgenes; four homozygous plants were identified from the two crossed lines tested. FISH results showing presence of transgenes were consistent with segregation ratios of expression of both transgenes, indicating that the two transgenes were expressed without transgene silencing in homozygous progeny advanced to the F(3) and F(4) generations. Thus, even after crossing independently transformed, homozygous parental plants containing a single, stably expressed transgene, progeny were obtained that continued to express multiple transgenes through generation advance. Such stability of transgenes, following outcrossing, is an important attribute for trait modification and for gene flow studies.

  1. Improving expression of reporter transgene in stem cell by construction of different lentiviral vectors

    Energy Technology Data Exchange (ETDEWEB)

    Tae, Seong Ho; Min, Jung Joon [Chonnam National University Medical School, Gwangju (Korea, Republic of); Le, Uyenchi N.; Padmanabhan, Parasuraman [Singapore Bio-Imaging Imaging Consortium, Singapore (Singapore)

    2007-07-01

    For stem cell trafficking applications, it is imperative to express transgenes at desired and stable levels. In recent years, lentivirus-mediated gene transfer was shown to be an efficient method to stably introduce genetic modifications in target cells, even if these are in proliferative or nonproliferative states. Moreover, transgene expression levels can be controlled by using different promoters. The present study was designed to compare the potency of various promoters regulating expression of imaging reporter genes in embryonic H9c2 cardiomyoblasts derived from rat heart. Lentiviral vector was produced by the transient transfection of plasmids carrying required genes and those encoding for virus coating proteins into 293T cells. Harvested viral constructs were incubated with Hela and H9c2 cells, respectively. Transgene expressions were detected by several imaging modalities and evaluated by enzymatic assays. Results - We observed that the level of stable transgene expression in lentivirus-transduced myoblasts could be modulated over several orders of magnitude, with the Ubiquitin (Ub) promoter exhibiting the highest activity, intermediate expression was observed with the CAG promoter, whereas expression observed with the CMV promoter was very weak. We observed that the level of stable transgene expression in lentivirus-transduced myoblasts could be modulated over several orders of magnitude, with the Ubiquitin (Ub) promoter exhibiting the highest activity, intermediate expression was observed with the CAG promoter, whereas expression observed with the CMV promoter was very weak. Here we show that lentivirus-mediated gene transfer allows efficient and stable transgene expression in embryonic cardiomyoblasts in vitro and that transgene expression levels can be varied by using different well-characterized gene promoters. In vivo trials about gene expression will probably further determine the potential of long-term trafficking stem cells using lentivirus.

  2. Improving expression of reporter transgene in stem cell by construction of different lentiviral vectors

    International Nuclear Information System (INIS)

    Tae, Seong Ho; Min, Jung Joon; Le, Uyenchi N.; Padmanabhan, Parasuraman

    2007-01-01

    For stem cell trafficking applications, it is imperative to express transgenes at desired and stable levels. In recent years, lentivirus-mediated gene transfer was shown to be an efficient method to stably introduce genetic modifications in target cells, even if these are in proliferative or nonproliferative states. Moreover, transgene expression levels can be controlled by using different promoters. The present study was designed to compare the potency of various promoters regulating expression of imaging reporter genes in embryonic H9c2 cardiomyoblasts derived from rat heart. Lentiviral vector was produced by the transient transfection of plasmids carrying required genes and those encoding for virus coating proteins into 293T cells. Harvested viral constructs were incubated with Hela and H9c2 cells, respectively. Transgene expressions were detected by several imaging modalities and evaluated by enzymatic assays. Results - We observed that the level of stable transgene expression in lentivirus-transduced myoblasts could be modulated over several orders of magnitude, with the Ubiquitin (Ub) promoter exhibiting the highest activity, intermediate expression was observed with the CAG promoter, whereas expression observed with the CMV promoter was very weak. We observed that the level of stable transgene expression in lentivirus-transduced myoblasts could be modulated over several orders of magnitude, with the Ubiquitin (Ub) promoter exhibiting the highest activity, intermediate expression was observed with the CAG promoter, whereas expression observed with the CMV promoter was very weak. Here we show that lentivirus-mediated gene transfer allows efficient and stable transgene expression in embryonic cardiomyoblasts in vitro and that transgene expression levels can be varied by using different well-characterized gene promoters. In vivo trials about gene expression will probably further determine the potential of long-term trafficking stem cells using lentivirus

  3. Bone Marrow Derived Mesenchymal Stromal Cells Harness Purinergenic Signaling to Tolerize Human Th1 Cells In Vivo

    Science.gov (United States)

    Amarnath, Shoba; Foley, Jason E.; Farthing, Don E.; Gress, Ronald E.; Laurence, Arian; Eckhaus, Michael A.; Métais, Jean-Yves; Rose, Jeremy J.; Hakim, Frances T.; Felizardo, Tania C.; Cheng, Austin V.; Robey, Pamela G.; Stroncek, David E.; Sabatino, Marianna; Battiwalla, Minoo; Ito, Sawa; Fowler, Daniel H.; Barrett, Austin J.

    2014-01-01

    The use of bone marrow derived mesenchymal stromal cells (BMSC) in the treatment of alloimmune and autoimmune conditions has generated much interest, yet an understanding of the therapeutic mechanism remains elusive. We therefore explored immune modulation by a clinical-grade BMSC product in a model of human-into-mouse xenogeneic GVHD (x-GVHD) mediated by human CD4+ Th1 cells. BMSC reversed established, lethal x-GVHD through marked inhibition of Th1 cell effector function. Gene marking studies indicated BMSC engraftment was limited to the lung; further, there was no increase in regulatory T cells, thereby suggesting a paracrine mechanism of BMSC action. BMSC recipients had increased serum CD73 expressing exosomes that promoted adenosine accumulation ex vivo. Importantly, immune modulation mediated by BMSC was fully abrogated by pharmacologic therapy with an adenosine A2A receptor antagonist. To investigate the potential clinical relevance of these mechanistic findings, patient serum samples collected pre- and post-BMSC treatment were studied for exosome content: CD73 expressing exosomes promoting adenosine accumulation were detected in post-BMSC samples. In conclusion, BMSC effectively modulate experimental GVHD through a paracrine mechanism that promotes adenosine-based immune suppression. PMID:25532725

  4. Steps toward a globally available malaria vaccine: harnessing the potential of algae for future low cost vaccines.

    Science.gov (United States)

    Jones, Carla S; Mayfield, Stephen P

    2013-01-01

    Malaria is an infectious disease that threatens half of the world's population. This debilitating disease is caused by infection from parasites of the genus Plasmodium. Insecticides, bed nets and drug therapies have lowered the prevalence and death rate associated with malaria but this disease continues to plague many populations around the world. In recent years, many organizations have suggested developing methods for a complete eradication of malaria. The most straightforward and effective method for this potential eradication will be through the development of a low-cost vaccine. To achieve eradication, it will be necessary to develop new vaccine candidates and novel systems for both the production and delivery of these vaccines. Recently, the green algae Chlamydomonas reinhardtii has been used for the recombinant expression of malaria vaccine candidates including the transmission blocking vaccine candidate Pfs48/45. Here, we discuss the potential of this research on the future development of a low-cost malaria vaccine candidate.

  5. Harnessing Nanotopography to Enhance Osseointegration of Clinical Orthopedic Titanium Implants—An in Vitro and in Vivo Analysis

    Directory of Open Access Journals (Sweden)

    Vitali Goriainov

    2018-04-01

    Full Text Available Despite technological advancements, further innovations in the field of orthopedics and bone regeneration are essential to meet the rising demands of an increasing aging population and associated issues of disease, injury and trauma. Nanotopography provides new opportunities for novel implant surface modifications and promises to deliver further improvements in implant performance. However, the technical complexities of nanotopography fabrication and surface analysis have precluded identification of the optimal surface features to trigger osteogenesis. We herein detail the osteoinductive potential of discrete nanodot and nanowire nanotopographies. We have examined the ability of modified titanium and titanium alloy (Ti64 surfaces to induce bone-specific gene activation and extracellular matrix protein expression in human skeletal stem cells (SSCs in vitro, and de novo osteogenic response within a murine calvarial model in vivo. This study provides evidence of enhanced osteogenic response to nanowires 300 surface modifications, with important implications for clinical orthopedic application.

  6. Harnessing Integrative Omics to Facilitate Molecular Imaging of the Human Epidermal Growth Factor Receptor Family for Precision Medicine.

    Science.gov (United States)

    Pool, Martin; de Boer, H Rudolf; Hooge, Marjolijn N Lub-de; van Vugt, Marcel A T M; de Vries, Elisabeth G E

    2017-01-01

    Cancer is a growing problem worldwide. The cause of death in cancer patients is often due to treatment-resistant metastatic disease. Many molecularly targeted anticancer drugs have been developed against 'oncogenic driver' pathways. However, these treatments are usually only effective in properly selected patients. Resistance to molecularly targeted drugs through selective pressure on acquired mutations or molecular rewiring can hinder their effectiveness. This review summarizes how molecular imaging techniques can potentially facilitate the optimal implementation of targeted agents. Using the human epidermal growth factor receptor (HER) family as a model in (pre)clinical studies, we illustrate how molecular imaging may be employed to characterize whole body target expression as well as monitor drug effectiveness and the emergence of tumor resistance. We further discuss how an integrative omics discovery platform could guide the selection of 'effect sensors' - new molecular imaging targets - which are dynamic markers that indicate treatment effectiveness or resistance.

  7. Saponin determination, expression analysis and functional characterization of saponin biosynthetic genes in Chenopodium quinoa leaves.

    Science.gov (United States)

    Fiallos-Jurado, Jennifer; Pollier, Jacob; Moses, Tessa; Arendt, Philipp; Barriga-Medina, Noelia; Morillo, Eduardo; Arahana, Venancio; de Lourdes Torres, Maria; Goossens, Alain; Leon-Reyes, Antonio

    2016-09-01

    Quinoa (Chenopodium quinoa Willd.) is a highly nutritious pseudocereal with an outstanding protein, vitamin, mineral and nutraceutical content. The leaves, flowers and seed coat of quinoa contain triterpenoid saponins, which impart bitterness to the grain and make them unpalatable without postharvest removal of the saponins. In this study, we quantified saponin content in quinoa leaves from Ecuadorian sweet and bitter genotypes and assessed the expression of saponin biosynthetic genes in leaf samples elicited with methyl jasmonate. We found saponin accumulation in leaves after MeJA treatment in both ecotypes tested. As no reference genes were available to perform qPCR in quinoa, we mined publicly available RNA-Seq data for orthologs of 22 genes known to be stably expressed in Arabidopsis thaliana using geNorm, NormFinder and BestKeeper algorithms. The quinoa ortholog of At2g28390 (Monensin Sensitivity 1, MON1) was stably expressed and chosen as a suitable reference gene for qPCR analysis. Candidate saponin biosynthesis genes were screened in the quinoa RNA-Seq data and subsequent functional characterization in yeast led to the identification of CqbAS1, CqCYP716A78 and CqCYP716A79. These genes were found to be induced by MeJA, suggesting this phytohormone might also modulate saponin biosynthesis in quinoa leaves. Knowledge of the saponin biosynthesis and its regulation in quinoa may aid the further development of sweet cultivars that do not require postharvest processing. Copyright © 2016 Elsevier Ireland Ltd. All rights reserved.

  8. Real-time quantitative reverse transcription-PCR analysis of expression stability of Actinobacillus pleuropneumoniae housekeeping genes during in vitro growth under iron-depleted conditions

    DEFF Research Database (Denmark)

    Nielsen, K. K.; Boye, Mette

    2005-01-01

    up-regulation under iron-restricted conditions compared to bacteria grown in medium with sufficient iron. The observed expression patterns of the genes of interest were consistent with previous observations. This study therefore lends further support to the use of real-time quantitative RT...... controls, as such controls have not been defined yet for this bacterium. Bacterial gene expression was studied during in vitro exponential and early stationary growth in medium with and without sufficient iron, respectively. First, the stability of expression of five genes, the glyA, tpiA, pykA, rec......F, and rhoAP genes involved in basic housekeeping, was evaluated on the basis of the mean pairwise variation. All the housekeeping genes included were stably expressed under the conditions investigated and consequently were included in the normalization procedure. Next, the geometric mean of the internal...

  9. Recovery of NIS expression in thyroid cancer cells by overexpression of Pax8 gene

    International Nuclear Information System (INIS)

    Presta, Ivan; Filetti, Sebastiano; Russo, Diego; Arturi, Franco; Ferretti, Elisabetta; Mattei, Tiziana; Scarpelli, Daniela; Tosi, Emanuele; Scipioni, Angela; Celano, Marilena; Gulino, Alberto

    2005-01-01

    Recovery of iodide uptake in thyroid cancer cells by means of obtaining the functional expression of the sodium/iodide symporter (NIS) represents an innovative strategy for the treatment of poorly differentiated thyroid cancer. However, the NIS gene expression alone is not always sufficient to restore radioiodine concentration ability in these tumour cells. In this study, the anaplastic thyroid carcinoma ARO cells were stably transfected with a Pax8 gene expression vector. A quantitative RT-PCR was performed to assess the thyroid specific gene expression in selected clones. The presence of NIS protein was detected by Western blot and localized by immunofluorescence. A iodide uptake assay was also performed to verify the functional effect of NIS induction and differentiation switch. The clones overexpressing Pax8 showed the re-activation of several thyroid specific genes including NIS, Pendrin, Thyroglobulin, TPO and TTF1. In ARO-Pax8 clones NIS protein was also localized both in cell cytoplasm and membrane. Thus, the ability to uptake the radioiodine was partially restored, associated to a high rate of efflux. In addition, ARO cells expressing Pax8 presented a lower rate of cell growth. These finding demonstrate that induction of Pax8 expression may determine a re-differentiation of thyroid cancer cells, including a partial recovery of iodide uptake, fundamental requisite for a radioiodine-based therapeutic approach for thyroid tumours

  10. Clearing the corpses: regulatory mechanisms, novel tools, and therapeutic potential of harnessing microglial phagocytosis in the diseased brain

    Directory of Open Access Journals (Sweden)

    Irune Diaz-Aparicio

    2016-01-01

    Full Text Available Apoptosis is a widespread phenomenon that occurs in the brain in both physiological and pathological conditions. Dead cells must be quickly removed to avoid the further toxic effects they exert in the parenchyma, a process executed by microglia, the brain professional phagocytes. Although phagocytosis is critical to maintain tissue homeostasis, it has long been either overlooked or indirectly assessed based on microglial morphology, expression of classical activation markers, or engulfment of artificial phagocytic targets in vitro. Nevertheless, these indirect methods present several limitations and, thus, direct observation and quantification of microglial phagocytosis is still necessary to fully grasp its relevance in the diseased brain. To overcome these caveats and obtain a comprehensive, quantitative picture of microglial phagocytosis we have developed a novel set of parameters. These parameters have allowed us to identify the different strategies utilized by microglia to cope with apoptotic challenges induced by excitotoxicity or inflammation. In contrast, we discovered that in mouse and human epilepsy microglia failed to find and engulf apoptotic cells, resulting in accumulation of debris and inflammation. Herein, we advocate that the efficiency of microglial phagocytosis should be routinely tested in neurodegenerative and neurological disorders, in order to determine the extent to which it contributes to apoptosis and inflammation found in these conditions. Finally, our findings point towards enhancing microglial phagocytosis as a novel therapeutic strategy to control tissue damage and inflammation, and accelerate recovery in brain diseases.

  11. Harnessing the incretin system beyond glucose control: potential cardiovascular benefits of GLP-1 receptor agonists in type 2 diabetes.

    Science.gov (United States)

    Cariou, B

    2012-10-01

    The management of type 2 diabetes continues to evolve as new data emerge. Although glycaemic control is still important, other risk factors--such as hypertension, dyslipidaemia and obesity--must also be addressed in order to reduce the long-term risks of cardiovascular complications and mortality. In this context, targeting the incretin system, and glucagon-like peptide-1 (GLP-1) in particular, has generated much interest. GLP-1 is released from the gut in response to food ingestion and plays a crucial role in glucose homeostasis. GLP-1 receptors are expressed in the heart and vasculature, prompting evaluation of their physiological role and pharmacological stimulation, both in healthy and disease states. These studies indicate that GLP-1 and GLP-1-based therapies appear to have direct, beneficial effects on the cardiovascular system, in addition to their glucose-lowering properties, such as modulation of blood pressure, endothelial function, and myocardial contractility. Intriguingly, some of these effects appear to be independent of GLP-1 receptor signalling. Data from clinical studies of the GLP-1 receptor agonists, exenatide and liraglutide on cardiovascular risk factors, in patients with type 2 diabetes are also promising and the results from prospective studies to assess cardiovascular outcomes are eagerly awaited. Copyright © 2012 Elsevier Masson SAS. All rights reserved.

  12. Multiple Sclerosis and the LIF/IL-6 Axis: Use of Nanotechnology to Harness the Tolerogenic and Reparative Properties of LIF

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    Su M. Metcalfe

    2015-05-01

    Full Text Available Leukaemia inhibitory factor (LIF plays a critical role in “stemness” versus “differentiation”, a property that underpins the core value of LIF as a therapeutic for both the treatment of autoimmune disease and for promoting tissue repair. This value can be realized using nano-engineering technology, where a new generation of tools can, with unprecedented ability, manipulate biological functions. One striking example is the treatment of multiple sclerosis (MS. The underpinning biology is the newly identified LIF/IL-6 axis in T lymphocytes, which can tilt the behaviour between immune tolerance versus immune attack. This LIF/IL-6 axis is ideally suited to nanotherapeutic manipulation, given its inherent mechanistic simplicity of two mutually opposing feed-forward loops that determine either tolerogenic (LIF or inflammatory (IL-6 immunity. Using LIF that is formulated in biodegradable nanoparticles (LIF-NP and targeted to CD4+ T cells, the axis is harnessed towards immune tolerance. This has implications for the treatment of autoimmune diseases, where the clinical burden is immense. It encompasses more than 100 diseases and, in the USA alone, costs more than $100 billion in direct health care costs annually. Other properties of LIF include the promotion of healthy neuro-glial interactions within the central nervous system (CNS, where, in addition to MS, LIF-NP therapy is relevant to inflammatory neurodegenerative diseases that represent a large and increasing need within aging populations. Thirdly, LIF is a reparative growth factor that can maintain genomic plasticity. LIF-NP supports the use of stem cell-based therapies in regenerative medicine plus augment therapeutic benefits within the patient. These core properties of LIF are greatly amplified in value by the advantage of being formulated as nanoparticles, namely (i targeted delivery, (ii exploitation of endogenous regulatory pathways and (iii creation of surrogate micro-stromal niches

  13. Expression Stabilities of Candidate Reference Genes for RT-qPCR in Chinese Jujube (Ziziphus jujuba Mill. under a Variety of Conditions.

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    Jiaodi Bu

    Full Text Available Reverse transcription-quantitative real-time polymerase chain reaction (RT-qPCR is a powerful method for evaluating patterns of gene expression. Jujube whole-genome sequencing has been completed, and analysis of gene function, an important part of any follow-up study, requires the appropriate selection of reference genes. Indeed, suitable reference gene selection for RT-qPCR is critical for accurate normalization of target gene expression. In this study, the software packages geNorm and NormFinder were employed to examine the expression stabilities of nine candidate reference genes under a variety of conditions. Actin-depolymerizing factor 1 (ACT1, Histone-H3 (His3, and Polyadenylate-binding protein-interacting protein (PAIP were determined to be the most stably expressed genes during five stages of fruit development and ACT1, SiR-Fd, BTF3, and Tubulin alpha chain (TUA across different tissues/organs. Whereas ACT1, Basic Transcription factor 3 (BTF3, Glyceraldehyde-3-phosphate dehydrogenase (GADPH, and PAIP were the most stable under dark conditions. ACT1, PAIP, BTF3, and Elongation factor 1- gamma (EF1γ were the most stably expressed genes under phytoplasma infection. Among these genes, SiR-Fd and PAIP are here first reported as stable reference genes. When normalized using these most stable reference genes, the expression patterns of four target genes were found to be in accordance with physiological data, indicating that the reference genes selected in our study are suitable for use in such analyses. This study provides appropriate reference genes and corresponding primers for further RT-qPCR studies in Chinese jujube and emphasizes the importance of validating reference genes for gene expression analysis under variable experimental conditions.

  14. Comparison of internal ribosome entry site (IRES and Furin-2A (F2A for monoclonal antibody expression level and quality in CHO cells.

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    Steven C L Ho

    Full Text Available Four versions of tricistronic vectors expressing IgG1 light chain (LC, IgG1 heavy chain (HC, and dihydrofolate reductase (DHFR in one transcript were designed to compare internal ribosome entry site (IRES and furin-2A (F2A for their influence on monoclonal antibody (mAb expression level and quality in CHO DG44 cells. LC and HC genes are arranged as either the first or the second cistron. When using mAb quantification methods based on the detection antibodies against HC Fc region, F2A-mediated tricistronic vectors appeared to express mAb at higher levels than the IRES-mediated tricistronic vectors in both transient and stable transfections. Further analysis revealed that more than 40% of products detected in stably transfected pools generated using the two F2A-mediated tricistronic vectors were aggregates. LC and HC from the F2A stably transfected pools were not properly processed, giving rise to LC+F2A+HC or HC+F2A+LC fusion proteins, LC and HC polypeptides with F2A remnants, and incorrectly cleaved signal peptides. Both IRES-mediated tricistronic vectors express mAb with correct sizes and signal peptide cleavage. Arrangement of LC as the first cistron in the IRES-mediated tricistronic vectors exhibits increased mAb expression level, better growth, and minimized product aggregation, while arrangement of HC as first cistron results in low expression, slower growth, and high aggregation. The results obtained will be beneficial for designing vectors that enhance mAb expression level and quality in mammalian cells.

  15. An in vivo transfection system for inducible gene expression and gene silencing in murine hepatocytes.

    Science.gov (United States)

    Hubner, Eric K; Lechler, Christian; Kohnke-Ertel, Birgit; Zmoos, Anne-Flore; Sage, Julien; Schmid, Roland M; Ehmer, Ursula

    2017-01-01

    Hydrodynamic tail vein injection (HTVI) of transposon-based integration vectors is an established system for stably transfecting mouse hepatocytes in vivo that has been successfully employed to study key questions in liver biology and cancer. Refining the vectors for transposon-mediated hepatocyte transfection will further expand the range of applications of this technique in liver research. In the present study, we report an advanced transposon-based system for manipulating gene expression in hepatocytes in vivo. Transposon-based vector constructs were generated to enable the constitutive expression of inducible Cre recombinase (CreER) together with tetracycline-inducible transgene or miR-small hairpin RNA (shRNA) expression (Tet-ON system). Transposon and transposase expression vectors were co-injected into R26R-mTmG reporter mice by HTVI. Cre-mediated gene recombination was induced by tamoxifen, followed by the administration of doxycycline to drive tetracycline-inducible gene or shRNA expression. Expression was visualized by immunofluorescence staining in livers of injected mice. After HTVI, Cre recombination by tamoxifen led to the expression of membrane-bound green fluorescent protein in transfected hepatocytes. Activation of inducible gene or shRNA expression was detected by immunostaining in up to one-third of transfected hepatocytes, with an efficiency dependent on the promoter driving the Tet-ON system. Our vector system combines Cre-lox mediated gene mutation with inducible gene expression or gene knockdown, respectively. It provides the opportunity for rapid and specific modification of hepatocyte gene expression and can be a useful tool for genetic screening approaches and analysis of target genes specifically in genetically engineered mouse models. Copyright © 2016 John Wiley & Sons, Ltd.

  16. Ubiquitin fusion expression and tissue-dependent targeting of hG-CSF in transgenic tobacco

    Science.gov (United States)

    2011-01-01

    Background Human granulocyte colony-stimulating factor (hG-CSF) is an important human cytokine which has been widely used in oncology and infection protection. To satisfy clinical needs, expression of recombinant hG-CSF has been studied in several organisms, including rice cell suspension culture and transient expression in tobacco leaves, but there was no published report on its expression in stably transformed plants which can serve as a more economical expression platform with potential industrial application. Results In this study, hG-CSF expression was investigated in transgenic tobacco leaves and seeds in which the accumulation of hG-CSF could be enhanced through fusion with ubiquitin by up to 7 fold in leaves and 2 fold in seeds, leading to an accumulation level of 2.5 mg/g total soluble protein (TSP) in leaves and 1.3 mg/g TSP in seeds, relative to hG-CSF expressed without a fusion partner. Immunoblot analysis showed that ubiquitin was processed from the final protein product, and ubiquitination was up-regulated in all transgenic plants analyzed. Driven by CaMV 35S promoter and phaseolin signal peptide, hG-CSF was observed to be secreted into apoplast in leaves but deposited in protein storage vacuole (PSV) in seeds, indicating that targeting of the hG-CSF was tissue-dependent in transgenic tobacco. Bioactivity assay showed that hG-CSF expressed in both seeds and leaves was bioactive to support the proliferation of NFS-60 cells. Conclusions In this study, the expression of bioactive hG-CSF in transgenic plants was improved through ubiquitin fusion strategy, demonstrating that protein expression can be enhanced in both plant leaves and seeds through fusion with ubiquitin and providing a typical case of tissue-dependent expression of recombinant protein in transgenic plants. PMID:21985646

  17. [Construction and clinical application of lentivirus-AQP4 expressing vector].

    Science.gov (United States)

    Liao, Zhang-yuan; Ye, Jing; Guan, Yun-qian; Zhang, Yu; You, Xiao-fan; An, Jing; Zhang, Ying; Wang, Shu-yan; Li, Cun-jiang

    2010-01-19

    To construct the human aquaporin-4 (AQP4) expressing vector and detect anti-AQP4 antibody in serum of patients with neuromyelitis optica (NMO). RNA was extracted from human glioblastoma and AQP4 cDNA obtained through RT-PCR.The fragment was cloned into the lentiviral expressing vector (iDUET101) and transformed into competent strain Hb101 for later amplification; plasmids were extracted from the amplified positive-bacteria-colony, sequenced and transfected into HEK-293T cells. Expression of AQP4 was identified by RT-PCR, Western blot and immunofluorescence assay. And anti-AQP4 antibody in human serum was tested. The sequence of target fragment matched with that of human AQP4 fragment sequences (NM_001650) completely. The constructed AQP4 fragment transfected in HEK-293T cell was tested by immunofluorescent examination and it exhibited obvious fluorescence located in cell membrane. Western blot test was positive. And the fragment was about 34 KD. Cellular immunofluorescence examination showed 11 examples of 12 NMO patient serums (91.7%) were positive, 4 in 34 multiple sclerosis (11.8%) positive and negative in all 50 serum samples of healthy controls. The HEK-293T cell transfected with lentivirus-AQP4 vector can express stably. And the expressed fragment may be applied in clinical examination.

  18. Photoacoustic imaging of gene expression using tyrosinase as a reporter gene

    Science.gov (United States)

    Paproski, Robert J.; Forbrich, Alexander; Harrison, Tyler; Hitt, Mary; Zemp, Roger J.

    2011-03-01

    Optical reporter genes, such as green fluorescence protein, are powerful research tools that allow visualization of gene expression. We have successfully used tyrosinase as a reporter gene for photoacoustic imaging. Tyrosinase is the key regulatory enzyme in the production of melanin which has a broad optical absorption spectrum. MCF-7 cells were stably transfected with tyrosinase under the control of an inducible promoter. For photoacoustic experiments, MCF-7 cells were resuspended at 108 cells/mL and injected in 700 μm (inner diameter) plastic tubing. Photoacoustic signal of MCF-7 cells expressing tyrosinase were >20-fold greater than those of untransfected MCF-7 cells. Photoacoustic signal of tyrosinaseexpressing MCF-7 cells were approximately 2-fold lesser and greater than those of blood at 576 and 650 nm, respectively, suggesting that photoacoustic signal from blood and tyrosinase-expressing cells can be separated by dualwavelength analysis. Photoacoustic signal from tyrosinase-expressing MCF-7 cells covered by chicken tissue could even be detected at a laser penetration depth of 4 cm, suggesting that tyrosinase can be used to image gene expression in relatively deep tissues. The current data suggests that tyrosinase is a strong reporter gene for photoacoustic imaging.

  19. Inducible expression of photoacoustic reporter gene tyrosinase in cells using a single plasmid

    Science.gov (United States)

    Paproski, Robert J.; Zemp, Roger J.

    2012-02-01

    We have previously demonstrated that tyrosinase is a reporter gene for photoacoustic imaging since tyrosinase is the rate-limiting step in the synthesis of melanin, a pigment capable of producing strong photoacoustic signals. We previously created a cell line capable of inducible tyrosinase expression (important due to toxicity of melanin) by stably transfecting tyrosinase in MCF-7 Tet-OnR cell line (Clontech) which expresses a doxycycline-controlled transactivator. Unfortunately, Clontech provides few Tet-On Advanced cell lines making it difficult to have inducible tyrosinase expression in cell lines not provided by Clontech. In order to simplify the creation of cell lines with inducible expression of tyrosinase, we created a single plasmid that encodes both the transactivator as well as tyrosinase. PCR was used to amplify both the transactivator and tyrosinase from the Tet-OnR Advanced and pTRE-Tight-TYR plasmids, respectively. Both PCR products were cloned into the pEGFP-N1 plasmid and the newly created plasmid was transfected into ZR-75-1, MCF-7, and MIA PaCa-1 cells using lipofectamine. After several days, brown melanin was only observed in cells incubated with doxycycline, suggesting that the newly created single plasmid allowed inducible tyrosinase expression in many different cells lines.

  20. The Construction and Expression of Lysine-Rich Gene in the Mammary Gland of Transgenic Mice

    Science.gov (United States)

    Ma, Xin; Zhang, Peng; Song, Guangqi; Chen, Yue; Wang, Zhongwei; Yin, Yupeng; Kong, Delong; Zhang, Sheng; Zhao, Zhihui; Ouyang, Hongsheng

    2012-01-01

    Lysine is the limiting amino acid in cereal grains, which represent a major source of human food and animal feed worldwide, and is considered the most important of the essential amino acids. In this study, β-casein, αS2-casein, and lactotransferrin cDNA clone fragments encoding lysine-rich peptides were fused together to generate a lysine-rich (LR) gene and the mammary gland-specific expression vector pBC1-LR-NEOr was constructed. Transgenic mice were generated by pronuclear microinjection of the linearized expression vectors harboring the LR transgene. The transgenic mice and their offspring were examined using multiplex polymerase chain reaction (PCR), Southern blotting, reverse transcriptase–PCR, in situ hybridization, and Western blotting techniques. Our results showed that the LR gene was successfully integrated into the mouse genome and was transmitted stably. The specific LR gene expression was restricted to the mammary gland, active alveoli of the transgenic female mice during lactation. The lysine level of the two transgenic lines was significantly higher than that of nontransgenic controls (ptransgenic pups was enhanced by directly feeding them the LR protein-enriched transgenic milk. Our results demonstrated that lysine-rich gene was successfully constructed and expressed in mammary gland of transgenic mice. This study will provide a better understanding of how mammary gland expression systems that increase the lysine content of milk can be applied to other mammals, such as cows. PMID:22577831

  1. With Reference to Reference Genes: A Systematic Review of Endogenous Controls in Gene Expression Studies.

    Science.gov (United States)

    Chapman, Joanne R; Waldenström, Jonas

    2015-01-01

    The choice of reference genes that are stably expressed amongst treatment groups is a crucial step in real-time quantitative PCR gene expression studies. Recent guidelines have specified that a minimum of two validated reference genes should be used for normalisation. However, a quantitative review of the literature showed that the average number of reference genes used across all studies was 1.2. Thus, the vast majority of studies continue to use a single gene, with β-actin (ACTB) and/or glyceraldehyde 3-phosphate dehydrogenase (GAPDH) being commonly selected in studies of vertebrate gene expression. Few studies (15%) tested a panel of potential reference genes for stability of expression before using them to normalise data. Amongst studies specifically testing reference gene stability, few found ACTB or GAPDH to be optimal, whereby these genes were significantly less likely to be chosen when larger panels of potential reference genes were screened. Fewer reference genes were tested for stability in non-model organisms, presumably owing to a dearth of available primers in less well characterised species. Furthermore, the experimental conditions under which real-time quantitative PCR analyses were conducted had a large influence on the choice of reference genes, whereby different studies of rat brain tissue showed different reference genes to be the most stable. These results highlight the importance of validating the choice of normalising reference genes before conducting gene expression studies.

  2. Leukocyte count affects expression of reference genes in canine whole blood samples

    Directory of Open Access Journals (Sweden)

    Dekker Aldo

    2011-02-01

    Full Text Available Abstract Background The dog is frequently used as a model for hematologic human diseases. In this study the suitability of nine potential reference genes for quantitative RT-PCR studies in canine whole blood was investigated. Findings The expression of these genes was measured in whole blood samples of 263 individual dogs, representing 73 different breeds and a group of 40 mixed breed dogs, categorized into healthy dogs and dogs with internal and hematological diseases, and dogs that underwent a surgical procedure. GeNorm analysis revealed that a combination of 5 to 6 of the most stably expressed genes constituted a stable normalizing factor. Evaluation of the expression revealed different ranking of reference genes in Normfinder and GeNorm. The disease category and the white blood cell count significantly affected reference gene expression. Conclusions The discrepancy between the ranking of reference genes in this study by Normfinder and Genorm can be explained by differences between the experimental groups such as "disease category" and "WBC count". This stresses the importance of assessing the expression stability of potential reference genes for gene experiments in canine whole blood anew for each specific experimental condition.

  3. Stable expression and characterization of human PN1 and PN3 sodium channels.

    Science.gov (United States)

    Akiba, Isamu; Seki, Tetsuo; Mori, Masayuki; Iizuka, Masaki; Nishimura, Seiichiro; Sasaki, Sachie; Imoto, Keiji; Barsoumian, Edward L

    2003-01-01

    Nociceptive transduction in inflammatory and neuropathic pain involves peripherally expressed voltage-gated sodium channels, such as tetrodotoxin (TTX)-sensitive PN1 and TTX-resistant PN3. We generated recombinant cell lines stably expressing the human PN1 and PN3 sodium channels in Chinese hamster ovary (CHO) cells using inducible expression vectors. The PN1 and PN3 cDNAs were isolated from human adrenal gland and heart poly(A)+ RNAs, respectively. The recombinant human PN1 currents exhibited rapid activation and inactivation kinetics and were blocked by TTX with a half-maximal inhibitory concentration (IC50) of 32.6 nM. The human PN3 channel expressed in stable transfectants showed TTX-resistant inward currents with slow inactivation kinetics. The IC50 value for TTX was 73.3 microM. The voltage-dependence of activation of the PN3 channel was shifted to the depolarizing direction, compared to that of the PN1 channel. Lidocaine and mexiletine exhibited tonic and use-dependent block of PN1 and PN3 channels. The PN1 channel was more susceptible to inhibition by mexiletine than PN3. These results suggest that stable transfectants expressing the human PN1 and PN3 sodium channels will be useful tools to define subtype selectivity for sodium channel blockers.

  4. A longitudinal study of gene expression in healthy individuals

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    Tessier Michel

    2009-06-01

    Full Text Available Abstract Background The use of gene expression in venous blood either as a pharmacodynamic marker in clinical trials of drugs or as a diagnostic test requires knowledge of the variability in expression over time in healthy volunteers. Here we defined a normal range of gene expression over 6 months in the blood of four cohorts of healthy men and women who were stratified by age (22–55 years and > 55 years and gender. Methods Eleven immunomodulatory genes likely to play important roles in inflammatory conditions such as rheumatoid arthritis and infection in addition to four genes typically used as reference genes were examined by quantitative reverse transcription-polymerase chain reaction (qRT-PCR, as well as the full genome as represented by Affymetrix HG U133 Plus 2.0 microarrays. Results Gene expression levels as assessed by qRT-PCR and microarray were relatively stable over time with ~2% of genes as measured by microarray showing intra-subject differences over time periods longer than one month. Fifteen genes varied by gender. The eleven genes examined by qRT-PCR remained within a limited dynamic range for all individuals. Specifically, for the seven most stably expressed genes (CXCL1, HMOX1, IL1RN, IL1B, IL6R, PTGS2, and TNF, 95% of all samples profiled fell within 1.5–2.5 Ct, the equivalent of a 4- to 6-fold dynamic range. Two subjects who experienced severe adverse events of cancer and anemia, had microarray gene expression profiles that were distinct from normal while subjects who experienced an infection had only slightly elevated levels of inflammatory markers. Conclusion This study defines the range and variability of gene expression in healthy men and women over a six-month period. These parameters can be used to estimate the number of subjects needed to observe significant differences from normal gene expression in clinical studies. A set of genes that varied by gender was also identified as were a set of genes with elevated

  5. Investigation of reference gene expression during human herpesvirus 6B infection indicates peptidylprolyl isomerase A as a stable reference gene and TATA box binding protein as a gene up-regulated by this virus.

    Science.gov (United States)

    Engdahl, Elin; Dunn, Nicky; Fogdell-Hahn, Anna

    2016-01-01

    When using relative gene expression for quantification of RNA it is crucial that the reference genes used for normalization do not change with the experimental condition. We aimed at investigating the expressional stability of commonly used reference genes during Human herpesvirus 6B (HHV-6B) infection. Expression of eight commonly used reference genes were investigated with quantitative PCR in a T-cell line infected with HHV-6B. The stability of genes was investigated using the 2(-ΔΔCT) method and the algorithms BestKeeper, GeNorm and NormFinder. Our results indicate that peptidylprolyl isomerase A (PPIA) is the most stably expressed gene while TATA box binding protein (TBP) is the least stably expressed gene during HHV-6B infection. In a confirmatory experiment, TBP was demonstrated to be dose and time dependently upregulated by HHV-6B. The stability of PPIA is in line with other studies investigating different herpesvirus infections whereas the finding that HHV-6B significantly upregulates TBP is novel and most likely specific to HHV-6B. Copyright © 2015 Elsevier B.V. All rights reserved.

  6. Suppression of the lipopolysaccharide-induced expression of MARCKS-related protein (MRP) affects transmigration in activated RAW264.7 cells.

    Science.gov (United States)

    Chun, Kwang-Rok; Bae, Eun Mi; Kim, Jae-Kwan; Suk, Kyoungho; Lee, Won-Ha

    2009-01-01

    The molecular action mechanism of MRP, one of the protein kinase C (PKC) substrates, has been under intense investigation, but reports on its role in macrophage function remain controversial. The treatment of macrophage cell lines with bacterial lipopolysaccharide (LPS) induced a high level of MRP expression suggesting that MRP plays a role in the function of activated macrophages. In order to investigate the role of MRP in activated RAW264.7 cells, we stably transfected MRP-specific shRNA expression constructs and tested for alterations in macrophage-related functions. The down-regulation of MRP expression resulted in a marked reduction in chemotaxis toward MCP-1 or extracellular matrix proteins. Furthermore, pharmacological inhibitors of PKC significantly inhibited the chemotaxis in RAW264.7 cells. These data reveals the pivotal role of MRP in the transmigration of activated RAW264.7 cells.

  7. The endogenous retroviral insertion in the human complement C4 gene modulates the expression of homologous genes by antisense inhibition.

    Science.gov (United States)

    Schneider, P M; Witzel-Schlömp, K; Rittner, C; Zhang, L

    2001-02-01

    Intron 9 contains the complete endogenous retrovirus HERV-K(C4) as a 6.4-kb insertion in 60% of human C4 genes. The retroviral insertion is in reverse orientation to the C4 coding sequence. Therefore, expression of C4 could lead to the transcription of an antisense RNA, which might protect against exogenous retroviral infections. To test this hypothesis, open reading frames from the HERV sequence were subcloned in sense orientiation into a vector allowing expression of a beta-galactosidase fusion protein. Mouse L cells which had been stably transfected with either the human C4A or C4B gene both carrying the HERV insertion (LC4 cells), and L(Tk-) cells without the C4 gene were transiently transfected either with a retroviral construct or with the wild-type vector. Expression was monitored using an enzymatic assay. We demonstrated that (1) HERV-K(C4) antisense mRNA transcripts are present in cells constitutively expressing C4, (2) expression of retroviral-like constructs is significantly downregulated in cells expressing C4, and (3) this downregulation is further modulated in a dose-dependent fashion following interferon-gamma stimulation of C4 expression. These results support the hypothesis of a genomic antisense strategy mediated by the HERV-K(C4) insertion as a possible defense mechanism against exogenous retroviral infections.

  8. SATB1 regulates SPARC expression in K562 cell line through binding to a specific sequence in the third intron

    International Nuclear Information System (INIS)

    Li, K.; Cai, R.; Dai, B.B.; Zhang, X.Q.; Wang, H.J.; Ge, S.F.; Xu, W.R.; Lu, J.

    2007-01-01

    Special AT-rich binding protein 1 (SATB1), a cell type-specific nuclear matrix attachment region (MAR) DNA-binding protein, tethers to a specific DNA sequence and regulates gene expression through chromatin remodeling and HDAC (histone deacetylase complex) recruitment. In this study, a SATB1 eukaryotic expression plasmid was transfected into the human erythroleukemia K562 cell line and individual clones that stably over-expressed the SATB1 protein were isolated. Microarray analysis revealed that hundreds of genes were either up- or down-regulated in the SATB1 over-expressing K562 cell lines. One of these was the extra-cellular matrix glycoprotein, SPARC (human secreted protein acidic and rich in cysteine). siRNA knock-down of SATB1 also reduced SPARC expression, which was consistent with elevated SPARC levels in the SATB1 over-expressing cell line. Bioinformatics software Mat-inspector showed that a 17 bp DNA sequence in the third intron of SPARC possessed a high potential for SATB1 binding; a finding confirmed by Chromatin immunoprecipitation (ChIP) with anti-SATB1 antibody. Our results show for the first time that forced-expression of SATB1 in K562 cells triggers SPARC up-regulation by binding to a 17 bp DNA sequence in the third intron

  9. A construct with fluorescent indicators for conditional expression of miRNA

    Directory of Open Access Journals (Sweden)

    Xia Xugang

    2008-10-01

    -mediated induction of the miRNA expression. This construct can be used to increase the efficiency of making cell lines or transgenic animals that stably express miRNA targeting specific genes.

  10. Identification and validation of superior reference gene for gene expression normalization via RT-qPCR in staminate and pistillate flowers of Jatropha curcas - A biodiesel plant.

    Science.gov (United States)

    Karuppaiya, Palaniyandi; Yan, Xiao-Xue; Liao, Wang; Wu, Jun; Chen, Fang; Tang, Lin

    2017-01-01

    Physic nut (Jatropha curcas L) seed oil is a natural resource for the alternative production of fossil fuel. Seed oil production is mainly depended on seed yield, which was restricted by the low ratio of staminate flowers to pistillate flowers. Further, the mechanism of physic nut flower sex differentiation has not been fully understood yet. Quantitative Real Time-Polymerase Chain Reaction is a reliable and widely used technique to quantify the gene expression pattern in biological samples. However, for accuracy of qRT-PCR, appropriate reference gene is highly desirable to quantify the target gene level. Hence, the present study was aimed to identify the stable reference genes in staminate and pistillate flowers of J. curcas. In this study, 10 candidate reference genes were selected and evaluated for their expression stability in staminate and pistillate flowers, and their stability was validated by five different algorithms (ΔCt, BestKeeper, NormFinder, GeNorm and RefFinder). Resulting, TUB and EF found to be the two most stably expressed reference for staminate flower; while GAPDH1 and EF found to be the most stably expressed reference gene for pistillate flowers. Finally, RT-qPCR assays of target gene AGAMOUS using the identified most stable reference genes confirmed the reliability of selected reference genes in different stages of flower development. AGAMOUS gene expression levels at different stages were further proved by gene copy number analysis. Therefore, the present study provides guidance for selecting appropriate reference genes for analyzing the expression pattern of floral developmental genes in staminate and pistillate flowers of J. curcas.

  11. Identification and validation of superior reference gene for gene expression normalization via RT-qPCR in staminate and pistillate flowers of Jatropha curcas - A biodiesel plant.

    Directory of Open Access Journals (Sweden)

    Palaniyandi Karuppaiya

    Full Text Available Physic nut (Jatropha curcas L seed oil is a natural resource for the alternative production of fossil fuel. Seed oil production is mainly depended on seed yield, which was restricted by the low ratio of staminate flowers to pistillate flowers. Further, the mechanism of physic nut flower sex differentiation has not been fully understood yet. Quantitative Real Time-Polymerase Chain Reaction is a reliable and widely used technique to quantify the gene expression pattern in biological samples. However, for accuracy of qRT-PCR, appropriate reference gene is highly desirable to quantify the target gene level. Hence, the present study was aimed to identify the stable reference genes in staminate and pistillate flowers of J. curcas. In this study, 10 candidate reference genes were selected and evaluated for their expression stability in staminate and pistillate flowers, and their stability was validated by five different algorithms (ΔCt, BestKeeper, NormFinder, GeNorm and RefFinder. Resulting, TUB and EF found to be the two most stably expressed reference for staminate flower; while GAPDH1 and EF found to be the most stably expressed reference gene for pistillate flowers. Finally, RT-qPCR assays of target gene AGAMOUS using the identified most stable reference genes confirmed the reliability of selected reference genes in different stages of flower development. AGAMOUS gene expression levels at different stages were further proved by gene copy number analysis. Therefore, the present study provides guidance for selecting appropriate reference genes for analyzing the expression pattern of floral developmental genes in staminate and pistillate flowers of J. curcas.

  12. Gene expression analysis in prostate cancer: the importance of the endogenous control.

    Science.gov (United States)

    Vajda, Alice; Marignol, Laure; Barrett, Ciara; Madden, Stephen F; Lynch, Thomas H; Hollywood, Donal; Perry, Antoinette S

    2013-03-01

    Aberrant gene expression is a hallmark of cancer. Quantitative reverse-transcription PCR (qRT-PCR) is the gold-standard for quantifying gene expression, and commonly employs a house-keeping gene (HKG) as an endogenous control to normalize results; the choice of which is critical for accurate data interpretation. Many factors, including sample type, pathological state, and oxygen levels influence gene expression including putative HKGs. The aim of this study was to determine the suitability of commonly used HKGs for qRT-PCR in prostate cancer. Prostate cancer (LNCaP, 22Rv1, PC3, and DU145) and normal (PWR1E and RWPE1) cell lines were cultured in air and hypoxia. The performance of 16 HKGs was assessed using Normfinder and coefficient of variation. In silico promoter analysis was performed to identify putative hypoxia response elements (HREs). The impact of the endogenous control on expression levels of HIF1A and GSTP1 was investigated by qRT-PCR in cell lines and tissue specimens respectively. Hypoxia altered expression of several HKGs: IPO8, B2M, and PGK1. The most stably expressed HKGs were ACTB, PPIA, and UBC. Both UBC and ACTB showed constitutive expression of HIF1A in air and hypoxia, while PGK1 falsely implied a sixfold hypoxia-induced down-regulation. In prostate tumors, UBC and PGK1 both revealed down-regulation of GSTP1 relative to matched benign, whereas ACTB showed variability. This study demonstrates that no universal endogenous control exists for gene expression studies, even within one disease type. It highlights the importance of validating expression of intended HKGs between different sample types and environmental exposures. Copyright © 2012 Wiley Periodicals, Inc.

  13. Maximal mixing rate in turbulent stably stratified Couette flow

    Science.gov (United States)

    Caulfield, C. P.; Kerswell, R. R.

    2001-11-01

    A rigorous upper bound on the long-time-averaged vertical buoyancy flux is derived from the Navier-Stokes equations for a Boussinesq fluid confined between two parallel horizontal plates a distance d apart, maintained at a constant statically stabilizing temperature difference Δ T and driven at a constant relative velocity Δ U. The upper bound on the volume and long-time-averaged vertical buoyancy flux \\cal B := limt arrow ∞ 1/t int^t0 g/ρ0 dtildet is \\cal B <= \\cal B_max=(1-16√2/Re) (Δ U)^3 /(64 √2d) where Re=Δ Ud/ν and ρ0 is some reference density. Significantly, \\cal B_max is independent of the bulk Richardson number of the flow and is achieved by an optimal solution with a mixing efficiency (or flux Richardson number) which approaches 0.5 as the Reynolds number becomes large. The time-averaged turbulent dissipation of kinetic energy and the time-averaged vertical buoyancy flux are then in equipartition for the optimising flow.

  14. Når skolens behov blir viktigere enn formell kompetanse: Hvordan veilede nyutdannede lærere til å undervise i skolefag de ikke har?

    Directory of Open Access Journals (Sweden)

    Merete Føinum

    2016-04-01

    Full Text Available Studien undersøker veiledning av nyutdannede lærere ved en norsk ungdomsskole. Artikkelen problematiserer de ekstra krevende arbeidsbetingelsene som disse nye lærerne møtte i sitt første år, da de ble satt til å undervise i skolefag de ikke hadde studert. Forskningsspørsmålet er: Hvordan fremstiller nyutdannede ungdomsskolelærere sine opplevelser med å undervise i skolefag de ikke selv er formelt kvalifisert i, og på hvilke måter kan veiledning bidra med støtte? Artikkelen presenterer utdrag fra veiledningssamtaler som eksemplifiserer to nye læreres fremstilling av eget arbeid med elevvurdering når de mangler formell kvalifikasjon i faget. De nyutdannedes perspektiv, og mentorens, fremheves som et samspill gjennom inter­aksjonsanalyse. Artikkelens teoretiske bidrag er analysebegrepene – «psykologisk-emosjonell mentorstøtte» og «faglig-profesjonell mentorstøtte» – som brukes for å identifisere ulike varianter av mentorstøtte i veiledningssamtaler. Innenfor hovedtypen «faglig-profesjonell mentorstøtte» utdypes distinksjonen videre når det skilles analytisk mellom mentor-bidrag som vektlegger spisset drøfting av skolefag (faglig og mentor-innspill av pedagogisk eller mer generell didaktisk karakter (profesjonell. Dermed bidrar studien til å vise hvordan ulike kvaliteter i faglig-profesjonell lærerveiledning kan gjenkjennes i praksis. Funnene viser hvor avgjørende det er at mentor har kompetanse i det skolefaget det skal veiledes i, såfremt skoler ønsker å tilby nyutdannede lærere spisset veiledning i skolefag. Funnene har implikasjoner for skolelederes/-eieres tilrettelegging av arbeidsforhold og organiseringen av veilednings­støtte for nyutdannede lærere.Nøkkelord: nyutdannede (førsteårslærere, mentor(ing, veiledning, undervisningskompe­tanse, skolefag, ungdomsskolen, lærerutdanningAbstract Research on teachers’ professional development establishes the first year of teaching as a vulnerable phase

  15. Perceived quality of and access to care among poor urban women in Kenya and their utilization of delivery care: harnessing the potential of private clinics?

    Science.gov (United States)

    Fotso, Jean Christophe; Mukiira, Carol

    2012-09-01

    This paper uses data from a maternal health study carried out in 2006 in two slums of Nairobi, Kenya, to: describe perceptions of access to and quality of care among women living in informal settlements of Nairobi, Kenya; quantify the effects of women's perceived quality of, and access to, care on the utilization of delivery services; and draw policy implications regarding the delivery of maternal health services to the urban poor. Based on the results of the facility survey, all health facilities were classified as 'appropriate' or 'inappropriate'. The research was based on the premise that despite the poor quality of these maternal health facilities, their responsiveness to the socio-cultural and economic sensitivities of women would result in good perceptions and higher utilization by women. Our results show a pattern of women's good perceptions in terms of access to, and quality of, health care provided by the privately owned, sub-standard and often unlicensed clinics and maternity homes located within their communities. In the multivariate model, the association between women's perceptions of access to and quality of care, and delivery at these 'inappropriate' facilities remained strong, graded and in the expected direction. Women from the study area are seldom able to reach not-for-profit private providers of maternal health care services like missionary and non-governmental organization (NGO) clinics and hospitals. Against the backdrop of challenges faced by the public sector in health care provision, we recommend that the government should harness the potential of private clinics operating in urban, resource-deprived settings. First, the government should regulate private health facilities operating in urban slum settlements to ensure that the services they offer meet the acceptable minimum standards of obstetric care. Second, 'good' facilities should be given technical support and supplied with drugs and equipment.

  16. Graf Type-IV Hips Have a Higher Risk of Residual Acetabular Dysplasia at 1 Year of Age Following Successful Pavlik Harness Treatment for Developmental Hip Dysplasia.

    Science.gov (United States)

    Novais, Eduardo N; Sanders, Julia; Kestel, Lauryn A; Carry, Patrick M; Meyers, Mariana L

    2016-09-22

    We asked whether patient-specific factors and ultrasound (US) measurements of hip dysplasia severity at initial examination influence short-term residual acetabular dysplasia in patients successfully treated with Pavlik harness for developmental hip dysplasia. After IRB approval, 134 hips (84 patients) successfully treated by the Pavlik method between August 2011 and October 2014 with follow-up at 12 months of age were identified. Early successful treatment was defined as normal examination and US after approximately 12 weeks of Pavlik treatment. Multivariate linear and logistic regression models were used to identify factors associated with acetabular index (AI) measurements at 12 months as well as factors associated with an increase in AI between the 6- and 12-month timepoints (dysplastic progression). The study consisted of 134 hips (84 patients). The distribution of dysplastic, Barlow, and Ortolani hips was 44.8% (N=60), 30.6% (N=41), and 24.6% (N=33), respectively. The crude incidence of residual dysplasia at the 6-month visit (AI>30 degrees) was 11.7% (12/102). The incidence of residual dysplasia at the 12-month visit (AI>28 degrees) was 11.8% (13/111). Graf type was the only variable associated with AI values at the 12-month visit (mean difference: Graf type-IV-Other, 2.6; 95% confidence interval, 0.3-4.9; P=0.026). The risk of residual acetabular dysplasia after normal hip US following Pavlik treatment is not negligible. Radiographic surveillance is warranted to monitor and screen for dysplasia progression. Patients with dislocated Graf type-IV hips at diagnosis are at increased risk of residual acetabular dysplasia at 1 year after successful treatment with the Pavlik method. Level III-therapeutic study.

  17. Human Behavior and Environmental Sustainability: promoting a pro-environmental behavior by harnessing the social, psychological and physical influences of the built environment

    Science.gov (United States)

    Abusafieh, Shaden; Razem, Maiss

    2017-11-01

    Recently, technological advancements in the sustainable design field have allowed us to reduce the ecological impact of the built environment, to reduce consumption of non-renewable resources, to create healthy environments and in some cases may even rehabilitate the ecosystem. Nevertheless, several studies have been carried out showing that sustainable technology does not automatically lead to environmentally friendly behaviors in its users. Various environmental problems threaten environmental sustainability and many of these problems are rooted in human behavior. Unfortunately, there is a lack in studies which take into consideration the human behavior influences within a sustainable built environment. We believe that the built environment should be used to support human goals and requirements, but at the same time we should consider it as a context in which human values and behaviors are cultivated. This research aimed to help in promoting environmental sustainability by using architectural design in changing relevant human behavior toward an environmentally friendly behavior. In order to achieve this, the research adopted Environment-centered Approach to gain more acute perspective into the relationship between the physical environment and human behavior, focusing on social, psychological and physical influences of the built environment. It appears that environmental psychology's merits have high potential in changing behavior within the built environment. The research provides a systematic approach for selecting, assessing, evaluating the behaviors to be changed and the factors that determine them. Furthermore, this approach helps in choosing the best interventions that could be applied in built environment to encourage such a sustainable behavior. This study tried to construct an agenda for further researches to find particular architectural design elements and strategies that we can harness to develop a pro-environment human behavior.

  18. The effects of multiple UV exposures on HIV-LTR (long terminal repeat) expression

    International Nuclear Information System (INIS)

    Schreck, S.; Milton, J.; Panozzo, J.; Libertin, C.R.; Woloschak, G.E.; Loyola Univ., Maywood, IL

    1995-01-01

    Previous studies have shown that cellular stress agents such as UV radiation induce transcription from the long terminal repeat (LTR) of the human immunodeficiency virus (HIV). Using HeLa cells stably transfected with the HIV-LTR sequence, which transcriptionally drives the chloramphenicol acetyl transferase (CAT) reporter gene, we examined the effects of multiple exposures to UVC (254 nm) on HIV-LTR-CAT expression. Low doses (≤ 5 J m -2 ) had no effect on CAT expression, but up to 29-fold induction was observed with 10 J m -2 when cells were harvested 48 h after completion of the exposure. Little difference was noted in induction levels when cells were exposed to one 25 J m -2 dose, viable cells were harvested at 24 h, 48 h or 72 h, and cell lysates were assayed for CAT expression. Two sequential 12.5 J m -2 exposures, given 24 h apart, resulted in an additive effect on CAT expression; these two exposures produced CAT activity equivalent to that induced following a single 25 J m -2 dose. Our data suggest that HIV-LTR requires a specific threshold UV dose in order to elicit induction; a maximal induction dose is also evident; exposures higher than this maximal dose contribute no more to HIV-LTR induction in viable cells. (author)

  19. A recombinant canine distemper virus expressing a modified rabies virus glycoprotein induces immune responses in mice.

    Science.gov (United States)

    Li, Zhili; Wang, Jigui; Yuan, Daoli; Wang, Shuang; Sun, Jiazeng; Yi, Bao; Hou, Qiang; Mao, Yaping; Liu, Weiquan

    2015-06-01

    Canine distemper virus (CDV) and rabies virus (RV) are two important pathogens of the dog. CDV, a member of the morbillivirus genus, has shown promise as an expression vector. The glycoprotein from RV is a main contributor to protective immunity and capable of eliciting the production of virus-neutralizing antibodies. In this study, we recovered an attenuated strain of canine distemper virus and constructed a recombinant virus, rCDV-RV-G, expressing a modified (R333Q) rabies virus glycoprotein (RV-G) of RV Flury strain LEP. RV-G expression by the recombinant viruses was confirmed. Furthermore, G was proved to be incorporated into the surface of CDV particles. While replication of the recombinant virus was slightly reduced compared with the parental CDV, it stably expressed the RV-G over ten serial passages. Inoculation of mice induced specific neutralizing antibodies against both RV-G and CDV. Therefore, the rCDV-RV-G has the potential as a vaccine that may be used to control rabies virus infection in dogs and other animals.

  20. Regulation and expression of Lcr plasmid-mediated peptides in pesticinogenic Yersinia pestis

    International Nuclear Information System (INIS)

    Sample, A.K.

    1987-01-01

    It is shown in this thesis that cells of Lcr + , Pst - Y. pestis KIM are able to express Yops at levels comparable to that of Lcr + Yersinia pseudotuberculosis. Pulse-chase radiolabeling with 35 S-methionine was used to demonstrate that Lcr + , Pst + Y. pestis synthesized at least 11 distinct peptides during the low calcium response and that seven of the labeled peptides were rapidly degraded. These seven peptides were stably expressed in Lcr + , Pst - Y. pestis and were of identical molecular weights as the Yops expressed by that strain. Radiolabeled fragments of low molecular weight accumulated in the extracellular medium of Pst + cultures and were assumed to be stable degradation fragments derived from Yops. It was also shown that the set of stable peptides, including V antigen, were made during restriction by both Pst + and Pst - Y. pestis KIM and were located primarily within the cytoplasm. Those radiolabeled peptides which underwent proteolytic degradation in Pst + Y. pestis were localized to the outer membrane and extracellular medium in the Pst - strain. It is concluded that the failure of Lcr + , Pst + Y. pestis to express Yops is the result of post-translational degradation and is not a block in the synthesis of Yops

  1. Dienogest inhibits C-C motif chemokine ligand 20 expression in human endometriotic epithelial cells.

    Science.gov (United States)

    Mita, Shizuka; Nakakuki, Masanori; Ichioka, Masayuki; Shimizu, Yutaka; Hashiba, Masamichi; Miyazaki, Hiroyasu; Kyo, Satoru

    2017-07-01

    C-C motif chemokine ligand 20 is thought to contribute to the development of endometriosis by recruiting Th17 lymphocytes into endometriotic foci. The present study investigated the effects of dienogest, a progesterone receptor agonist used to treat endometriosis, on C-C motif chemokine ligand 20 expression by endometriotic cells. Effects of dienogest on mRNA expression and protein secretion of C-C motif chemokine ligand 20 induced by interleukin 1β were assessed in three immortalized endometriotic epithelial cell lines, parental cells (EMosis-CC/TERT1), and stably expressing human progesterone receptor isoform A (EMosis-CC/TERT1/PRA+) or isoform B (EMosis-CC/TERT1/PRA-/PRB+). Dienogest markedly inhibited interleukin 1β-stimulated C-C motif chemokine ligand 20 mRNA expression and protein secretion in EMosis-CC/TERT1/PRA-/PRB+, which was abrogated by the progesterone receptor antagonist RU486. In EMosis-CC/TERT1/PRA+, dienogest slightly inhibited C-C motif chemokine ligand 20 mRNA and protein. In EMosis-CC/TERT1, dienogest slightly inhibited C-C motif chemokine ligand 20 mRNA, but had no effect on C-C motif chemokine ligand 20 protein. Dienogest inhibited interleukin 1β-induced up-regulation of C-C motif chemokine ligand 20 in endometriotic epithelial cells, mainly mediated by progesterone receptor B. Copyright © 2017 Elsevier B.V. All rights reserved.

  2. Profile of differentially expressed genes mediated by the type III epidermal growth factor receptor mutation expressed in a small-cell lung cancer cell line

    DEFF Research Database (Denmark)

    Pedersen, M.W.; Andersen, Thomas Thykjær; Ørntoft, Torben Falck

    2001-01-01

    negative small-cell-lung cancer cell line, GLC3, stably transfected with the EGFRvIII gene in a Tet-On system. By comparison of mRNA levels in EGFRvIII-GLC3 with those of Tet-On-GLC3, it was found that the levels of mRNAs encoding several transcription factors (ATF-3, JunD, and c-Myb), cell adhesion......Previous studies have shown a correlation between expression of the EGF receptor type III mutation (EGFRvIII) and a more malignant phenotype of various cancers including: non-small-cell lung cancer, glioblastoma multiforme, prostate cancer and breast cancer. Thus, a detailed molecular genetic...... understanding of how the EGFRvIII contributes to the malignant phenotype is of major importance for future therapy. The GeneChip Hu6800Set developed by Affymetrix was used to identify changes in gene expression caused by the expression of EGFRvIII. The cell line selected for the study was an EGF receptor...

  3. Regulating the expression of therapeutic transgenes by controlled intake of dietary essential amino acids.

    Science.gov (United States)

    Chaveroux, Cédric; Bruhat, Alain; Carraro, Valérie; Jousse, Céline; Averous, Julien; Maurin, Anne-Catherine; Parry, Laurent; Mesclon, Florent; Muranishi, Yuki; Cordelier, Pierre; Meulle, Aline; Baril, Patrick; Do Thi, Anh; Ravassard, Philippe; Mallet, Jacques; Fafournoux, Pierre

    2016-07-01

    Widespread application of gene therapy will depend on the development of simple methods to regulate the expression of therapeutic genes. Here we harness an endogenous signaling pathway to regulate therapeutic gene expression through diet. The GCN2-eIF2α signaling pathway is specifically activated by deficiencies in any essential amino acid (EAA); EAA deficiency leads to rapid expression of genes regulated by ATF4-binding cis elements. We found that therapeutic genes under the control of optimized amino acid response elements (AAREs) had low basal expression and high induced expression. We applied our system to regulate the expression of TNFSF10 (TRAIL) in the context of glioma therapy and found that intermittent activation of this gene by EEA-deficient meals retained its therapeutic efficacy while abrogating its toxic effects on normal tissue. The GCN2-eIF2α pathway is expressed in many tissues, including the brain, and is highly specific to EAA deficiency. Our system may be particularly well suited for intermittent regulation of therapeutic transgenes over short or long time periods.

  4. Expression of estrogen receptor alpha with a Tet-off adenoviral system induces G0/G1 cell cycle arrest in SKBr3 breast cancer cells.

    Science.gov (United States)

    Peng, Jing; Jordan, V Craig

    2010-02-01

    Endocrine therapies targeting estrogen action are pivotal for the prevention and treatment of ER-positive breast cancers. Previous studies sought to recreate hormone responsiveness by the stable expression of ERalpha in the ER-negative MDA-MB-231 breast cancer cells. Paradoxically, estrogen inhibits breast cancer cell growth when an exogenous ERalpha is expressed. In this study, we have built on previous studies by developing a Tet-off adenoviral system to express ERalpha in the ER-negative SKBr3 breast cancer cells that over-express both EGFR and HER2. This system efficiently delivers ERalpha and the expression level of ERalpha is controlled by doxycycline in a concentration-dependent manner. The growth of SKBr3 was inhibited by ERalpha expression and further inhibited in the presence of 1 nM 17beta-estradiol. SKBr3 cells were arrested at G0/G1 cell cycle upon ERalpha expression, which corresponded to an increase of p21Cip1/Waf1, hypo-phosphorylation of pRb and decrease of E2F1. Estrogen also reduced EGFR and HER2 expression in SKBr3 cells after ERalpha was expressed. Given that estrogen-induced increase of p21Cip1/Waf1 and decrease of E2F1 was also observed in MDA-MB-231 cells stably transfected with ERalpha, our results suggest that a common pathway might be shared by different breast cancer cell lines whose growth is suppressed by ectopic ERalpha and estrogen.

  5. Ha-ras oncogene expression directed by a milk protein gene promoter: tissue specificity, hormonal regulation, and tumor induction in transgenic mice

    International Nuclear Information System (INIS)

    Andres, A.C.; Schoenenberger, C.A.; Groner, B.; Henninghausen, L.; LeMeur, M.; Gelinger, P.

    1987-01-01

    The activated human Ha-ras oncogene was subjected to the control of the promoter region of the murine whey acidic protein (Wap) gene, which is expressed in mammary epithelial cells in response to lactogenic hormones. The Wap-ras gene was stably introduced into the mouse germ line of five transgenic mice (one male and four females). Wap-ras expression was observed in the mammary glands of lactating females in two lines derived from female founders. The tissue-directed and hormone-dependent Wap expression was conferred on the Ha-ras oncogene. The signals governing Wap expression are located within 2.5 kilobases of 5' flanking sequence. The other two lines derived from female founders did not express the chimeric gene. In the line derived from the male founder the Wap-ras gene is integrated into the Y chromosome. Expression was found in the salivary gland of male animals only. After a long latency, Wap-ras-expressing mice developed tumors. The tumors arose in tissues expressing Wap-ras - i.e., mammary or salivary glands. Compared to the corresponding nonmalignant tissues, Wap-ras expression was enhanced in the tumors

  6. Intercellular adhesion molecule-1 expression by skeletal muscle cells augments myogenesis

    Energy Technology Data Exchange (ETDEWEB)

    Goh, Qingnian; Dearth, Christopher L.; Corbett, Jacob T. [Department of Kinesiology, The University of Toledo, Toledo, OH (United States); Pierre, Philippe [Centre d’Immunologie de Marseille-Luminy U2M, Aix-Marseille Université, Marseille (France); INSERM U631, Institut National de la Santé et Recherche Médicale, Marseille (France); CNRS UMR6102, Centre National de la Recherche Scientifique, Marseille (France); Chadee, Deborah N. [Department of Biological Sciences, The University of Toledo, Toledo, OH (United States); Pizza, Francis X., E-mail: Francis.Pizza@utoledo.edu [Department of Kinesiology, The University of Toledo, Toledo, OH (United States)

    2015-02-15

    We previously demonstrated that the expression of intercellular adhesion molecule-1 (ICAM-1) by skeletal muscle cells after muscle overload contributes to ensuing regenerative and hypertrophic processes in skeletal muscle. The objective of the present study is to reveal mechanisms through which skeletal muscle cell expression of ICAM-1 augments regenerative and hypertrophic processes of myogenesis. This was accomplished by genetically engineering C2C12 myoblasts to stably express ICAM-1, and by inhibiting the adhesive and signaling functions of ICAM-1 through the use of a neutralizing antibody or cell penetrating peptide, respectively. Expression of ICAM-1 by cultured skeletal muscle cells augmented myoblast–myoblast adhesion, myotube formation, myonuclear number, myotube alignment, myotube–myotube fusion, and myotube size without influencing the ability of myoblasts to proliferate or differentiate. ICAM-1 augmented myotube formation, myonuclear accretion, and myotube alignment through a mechanism involving adhesion-induced activation of ICAM-1 signaling, as these dependent measures were reduced via antibody and peptide inhibition of ICAM-1. The adhesive and signaling functions of ICAM-1 also facilitated myotube hypertrophy through a mechanism involving myotube–myotube fusion, protein synthesis, and Akt/p70s6k signaling. Our findings demonstrate that ICAM-1 expression by skeletal muscle cells augments myogenesis, and establish a novel mechanism through which the inflammatory response facilitates growth processes in skeletal muscle. - Highlights: • We examined mechanisms through which skeletal muscle cell expression of ICAM-1 facilitates events of in vitro myogenesis. • Expression of ICAM-1 by cultured myoblasts did not influence their ability to proliferate or differentiate. • Skeletal muscle cell expression of ICAM-1 augmented myoblast fusion, myotube alignment, myotube–myotube fusion, and myotube size. • ICAM-1 augmented myogenic processes through

  7. Construction of a heterologous gene expression system in the banana rhizobacterium strain GW-3 and its colonization ability.

    Science.gov (United States)

    Wang, Yuguang; Xia, Qiyu; Zhang, He; Lu, Xuehua; Sun, Jianbo; Zhang, Xin

    2014-03-01

    Rhizobacteria inhabiting the rhizosphere are beneficial to their host plants, and can potentially serve as biocontrol agents to control plant diseases. We isolated the rhizobacterium strain GW-3, which was the dominant bacterium in the rhizosphere soils of healthy banana plants. Then, we constructed an expression system with a kanamycin resistance gene to express a heterologous protein in GW-3. Using the green fluorescent protein gene as the reporter, we monitored expression of the heterologous protein by detecting fluorescence intensity and conducting western blot analyses. The standard fluorescence intensity of the recombinant strain reached 1,482 ± 3.49 RFU. To study the colonization ability of GW-3, we inoculated this bacterium into sterilized and unsterilized rhizosphere soils and monitored the bacterial population over 25 days. The populations of GW-3 in rhizosphere soils first increased, then decreased, and finally reached a balance. Laser scanning confocal microscope analyses of fluorescence in banana roots after inoculation with GW-3 confirmed that the recombinant GW-3 strain stably colonized banana root surfaces. Analyses of the bacterial population in unsterilized rhizosphere soils showed that the recombinant GW-3 strain was still the dominant bacterium in banana rhizosphere soils at 25 days after inoculation. Together, these results showed that this expression system can be used to express a heterologous protein at high levels in a dominant rhizobacterium. By incorporating relevant resistance genes into the expression system, this method could be used to genetically engineer GW-3 to control banana wilt disease.

  8. Staphylococcus epidermidis polysaccharide intercellular adhesin induces IL-8 expression in human astrocytes via a mechanism involving TLR2.

    LENUS (Irish Health Repository)

    Stevens, Niall T

    2009-03-01

    Staphylococcus epidermidis is an opportunistic biofilm-forming pathogen associated with neurosurgical device-related meningitis. Expression of the polysaccharide intercellular adhesin (PIA) on its surface promotes S. epidermidis biofilm formation. Here we investigated the pro-inflammatory properties of PIA against primary and transformed human astrocytes. PIA induced IL-8 expression in a dose- and\\/or time-dependent manner from U373 MG cells and primary normal human astrocytes. This effect was inhibited by depletion of N-acetyl-beta-d-glucosamine polymer from the PIA preparation with Lycopersicon esculentum lectin or sodium meta-periodate. Expression of dominant-negative versions of the TLR2 and TLR4 adaptor proteins MyD88 and Mal in U373 MG cells inhibited PIA-induced IL-8 production. Blocking IL-1 had no effect. PIA failed to induce IL-8 production from HEK293 cells stably expressing TLR4. However, in U373 MG cells which express TLR2, neutralization of TLR2 impaired PIA-induced IL-8 production. In addition to IL-8, PIA also induced expression of other cytokines from U373 MG cells including IL-6 and MCP-1. These data implicate PIA as an important immunogenic component of the S. epidermidis biofilm that can regulate pro-inflammatory cytokine production from human astrocytes, in part, via TLR2.

  9. Moderne bioenergi har store muligheder

    DEFF Research Database (Denmark)

    Larsen, Hans Hvidtfeldt; Kossmann, J.; Sønderberg Petersen, L.

    2003-01-01

    Bioenergi er energi, der stammer fra vedvarende kilder af biologisk oprindelse. Normalt bruges energiafgrøder dyrket specielt til formålet, eller biprodukter fra landbrug, skovbrug eller fiskeri. Eksempler på bioenergikilder er træbrændsel, bagasse(udpressede sukkerrør), organisk affald, biogas og...

  10. Har du sikret din alderdom?

    DEFF Research Database (Denmark)

    Kiens, Bente; Aagaard, Peter Gjerndrup

    2006-01-01

    Daglig fysisk aktivitet kan betragtes som indbetaling op din personlige helbredspension. Jo mere du sætter ind på kontoen, og jo oftere du gør det, jo større er sandsynligheden for, at du lever længere og får et sundere helbred. Og det er aldrig for sent at starte med indbetalingerne....

  11. Harnessing motivation to alleviate neglect

    Directory of Open Access Journals (Sweden)

    Charlotte eRussell

    2013-06-01

    Full Text Available The syndrome of spatial neglect results from the combination of a number of deficits in attention, with patients demonstrating both spatially lateralised and non-lateralised impairments. Previous reports have hinted that there may be a motivational component to neglect and that modulating this might alleviate some of the debilitating symptoms. Additionally, recent work on the effects of reward on attention in healthy participants has revealed improvements across a number of paradigms. As the primary deficit in neglect has been associated with attention, this evidence for reward’s effects is potentially important. However, until very recently there have been few empirical studies addressing this potential therapeutic avenue. Here we review the growing body of evidence that attentional impairments in neglect can be reduced by motivation, for example in the form of preferred music or anticipated monetary reward, and discuss the implications of this for treatments for these patients. Crucially these effects of positive motivation are not observed in all patients with neglect, suggesting that the consequences of motivation may relate to individual lesion anatomy. Given the key role of dopaminergic systems in motivational processes, we suggest that motivational stimulation might act as a surrogate for dopaminergic stimulation. In addition, we consider the relationship between clinical post stroke apathy and lack of response to motivation.

  12. Hvad har de gang i?

    DEFF Research Database (Denmark)

    Hansen, Niels-Henrik Møller

    Denne rapport er et resultat af et samarbejdet mellem Holbæk Kommune, repræsenteret ved Ungdommens Hus, og Center for Ungdomsforskning, CeFU, ved Roskilde Universitetscenter. Rapporten præsenterer og beskriver resultaterne af henholdsvis en række fokusgruppeinterviews med udsatte unge i Holbæk samt...

  13. Harnessing Theories for Tool Support

    DEFF Research Database (Denmark)

    Liu, Zhiming; Mencl, Vladimir; Ravn, Anders Peter

    2006-01-01

    Software development tools need to support more and more phases of the entire development process, because applications must be developed more correctly and efficiently. The tools therefore need to integrate sophisticated checkers, generators and transformations. A feasible approach to ensure high...... quality of such add-ins is to base them on sound formal foundations. In order to know where such add-ins will fit, we investigate the use of an existing successful commercial tool and identify suitable places for adding formally supported checking, transformation and generation modules. The paper...

  14. Harness the Energy of Collaboration

    Science.gov (United States)

    Lieberman, Ann; Miller, Lynne

    2016-01-01

    As they write about the Learning Communities standard in "Reach the Highest Standard in Professional Learning: Learning Communities," authors Ann Lieberman and Lynne Miller consider why communities have gained widespread attention and support and summarize the established research base about them. They also describe three contexts in…

  15. Klimaforandringerne har allerede ramt os

    DEFF Research Database (Denmark)

    Voigt, Steen

    2009-01-01

    Træernes pollensæson starter i dag flere uger tidligere end for 20 år siden, og mængden af pollen fra fx el og birk er øget markant. Planternes vækstsæson er blevet længere, og det markerer en historisk ændring af naturens tilstand. En ændring, der drives frem af de globale klimaforandringer. Ogs...

  16. Harnessing Theories for Tool Support

    DEFF Research Database (Denmark)

    Liu, Zhiming; Mencl, Vladimir; Ravn, Anders Peter

    2006-01-01

    quality of such add-ins is to base them on sound formal foundations. In order to know where such add-ins will fit, we investigate the use of an existing successful commercial tool and identify suitable places for adding formally supported checking, transformation and generation modules. The paper...

  17. Harnessing neuroplasticity for clinical applications.

    Science.gov (United States)

    Cramer, Steven C; Sur, Mriganka; Dobkin, Bruce H; O'Brien, Charles; Sanger, Terence D; Trojanowski, John Q; Rumsey, Judith M; Hicks, Ramona; Cameron, Judy; Chen, Daofen; Chen, Wen G; Cohen, Leonardo G; deCharms, Christopher; Duffy, Charles J; Eden, Guinevere F; Fetz, Eberhard E; Filart, Rosemarie; Freund, Michelle; Grant, Steven J; Haber, Suzanne; Kalivas, Peter W; Kolb, Bryan; Kramer, Arthur F; Lynch, Minda; Mayberg, Helen S; McQuillen, Patrick S; Nitkin, Ralph; Pascual-Leone, Alvaro; Reuter-Lorenz, Patricia; Schiff, Nicholas; Sharma, Anu; Shekim, Lana; Stryker, Michael; Sullivan, Edith V; Vinogradov, Sophia

    2011-06-01

    Neuroplasticity can be defined as the ability of the nervous system to respond to intrinsic or extrinsic stimuli by reorganizing its structure, function and connections. Major advances in the understanding of neuroplasticity have to date yielded few established interventions. To advance the translation of neuroplasticity research towards clinical applications, the National Institutes of Health Blueprint for Neuroscience Research sponsored a workshop in 2009. Basic and clinical researchers in disciplines from central nervous system injury/stroke, mental/addictive disorders, paediatric/developmental disorders and neurodegeneration/ageing identified cardinal examples of neuroplasticity, underlying mechanisms, therapeutic implications and common denominators. Promising therapies that may enhance training-induced cognitive and motor learning, such as brain stimulation and neuropharmacological interventions, were identified, along with questions of how best to use this body of information to reduce human disability. Improved understanding of adaptive mechanisms at every level, from molecules to synapses, to networks, to behaviour, can be gained from iterative collaborations between basic and clinical researchers. Lessons can be gleaned from studying fields related to plasticity, such as development, critical periods, learning and response to disease. Improved means of assessing neuroplasticity in humans, including biomarkers for predicting and monitoring treatment response, are needed. Neuroplasticity occurs with many variations, in many forms, and in many contexts. However, common themes in plasticity that emerge across diverse central nervous system conditions include experience dependence, time sensitivity and the importance of motivation and attention. Integration of information across disciplines should enhance opportunities for the translation of neuroplasticity and circuit retraining research into effective clinical therapies.

  18. Harnessing neuroplasticity for clinical applications

    Science.gov (United States)

    Sur, Mriganka; Dobkin, Bruce H.; O'Brien, Charles; Sanger, Terence D.; Trojanowski, John Q.; Rumsey, Judith M.; Hicks, Ramona; Cameron, Judy; Chen, Daofen; Chen, Wen G.; Cohen, Leonardo G.; deCharms, Christopher; Duffy, Charles J.; Eden, Guinevere F.; Fetz, Eberhard E.; Filart, Rosemarie; Freund, Michelle; Grant, Steven J.; Haber, Suzanne; Kalivas, Peter W.; Kolb, Bryan; Kramer, Arthur F.; Lynch, Minda; Mayberg, Helen S.; McQuillen, Patrick S.; Nitkin, Ralph; Pascual-Leone, Alvaro; Reuter-Lorenz, Patricia; Schiff, Nicholas; Sharma, Anu; Shekim, Lana; Stryker, Michael; Sullivan, Edith V.; Vinogradov, Sophia

    2011-01-01

    Neuroplasticity can be defined as the ability of the nervous system to respond to intrinsic or extrinsic stimuli by reorganizing its structure, function and connections. Major advances in the understanding of neuroplasticity have to date yielded few established interventions. To advance the translation of neuroplasticity research towards clinical applications, the National Institutes of Health Blueprint for Neuroscience Research sponsored a workshop in 2009. Basic and clinical researchers in disciplines from central nervous system injury/stroke, mental/addictive disorders, paediatric/developmental disorders and neurodegeneration/ageing identified cardinal examples of neuroplasticity, underlying mechanisms, therapeutic implications and common denominators. Promising therapies that may enhance training-induced cognitive and motor learning, such as brain stimulation and neuropharmacological interventions, were identified, along with questions of how best to use this body of information to reduce human disability. Improved understanding of adaptive mechanisms at every level, from molecules to synapses, to networks, to behaviour, can be gained from iterative collaborations between basic and clinical researchers. Lessons can be gleaned from studying fields related to plasticity, such as development, critical periods, learning and response to disease. Improved means of assessing neuroplasticity in humans, including biomarkers for predicting and monitoring treatment response, are needed. Neuroplasticity occurs with many variations, in many forms, and in many contexts. However, common themes in plasticity that emerge across diverse central nervous system conditions include experience dependence, time sensitivity and the importance of motivation and attention. Integration of information across disciplines should enhance opportunities for the translation of neuroplasticity and circuit retraining research into effective clinical therapies. PMID:21482550

  19. Lithium: for harnessing renewable energy

    Science.gov (United States)

    Bradley, Dwight; Jaskula, Brian W.

    2014-01-01

    Lithium, which has the chemical symbol Li and an atomic number of 3, is the first metal in the periodic table. Lithium has many uses, the most prominent being in batteries for cell phones, laptops, and electric and hybrid vehicles. Worldwide sources of lithium are broken down by ore-deposit type as follows: closed-basin brines, 58%; pegmatites and related granites, 26%; lithium-enriched clays, 7%; oilfield brines, 3%; geothermal brines, 3%; and lithium-enriched zeolites, 3% (2013 statistics). There are over 39 million tons of lithium resources worldwide. Of this resource, the USGS estimates there to be approximately 13 million tons of current economically recoverable lithium reserves. To help predict where future lithium supplies might be located, USGS scientists study how and where identified resources are concentrated in the Earth’s crust, and they use that knowledge to assess the likelihood that undiscovered resources also exist.

  20. Har dansk handyrsproduktion en fremtid?

    DEFF Research Database (Denmark)

    Andersen, Johnny Michael

    2013-01-01

    Lønsomheden i den danske handyrproduktion var lav i 2011. Nettooverskuddet pr. produceret handyr var således negativt for samtlige størrelsesgrupper. Der var dog et positivt dækningsbidrag for de fleste grupper, som indikerer, at det er hensigtsmæssigt at opretholde produktionen på kort sigt, men....... producerede handyr forbliver dog negativt for samtlige størrelsesgrupper....

  1. Vi har brug for skammere

    DEFF Research Database (Denmark)

    Holm, Claus

    2007-01-01

    Skammen er en væsentlig ingrediens i opdragelsen af børn, men det overser vi, for skam er et fyord i den pædagogiske verden. Interview med børnebogsforfatteren Lene Kaaberbøl om, hvorfor skam gør os menneskelige....

  2. Harnessing Computer Games in Education

    Science.gov (United States)

    Jong, Morris S. Y.; Shang, Junjie; Lee, Fong-Lok; Lee, Jimmy H. M.

    2008-01-01

    Besides the ability of making learning more interesting, educators and researchers have been exploring other pedagogical potentials of computer games. How to employ games for constructivist learning and teaching has become an attention in the field of education and game design in recent years. This article gives an introduction to game-based…

  3. Materials in harnessing solar power

    Indian Academy of Sciences (India)

    2018-04-06

    Apr 6, 2018 ... reducing the usage of battery [12]. Further, a suitable battery such as Li ion battery can be used as a power backup. As per the present-day energy storage device technology, the battery design should be in such a way that it can provide a balance among specific energy and power, and life, obtaining ...

  4. Arbejdsulykker! Alle har en historie

    DEFF Research Database (Denmark)

    Richter, Anne; Pedersen, Elsebet Frydendal

    in a trade school and in three carpentry companies in relation to five construction sites. The project draws on organizational culture theories of symbolic interactionism and on practice based learning i communities of practice. The methodological approach is primarily ethnographic. Four different cultures...

  5. Solar Radiation: Harnessing the Power

    Science.gov (United States)

    Rowland, Teri; Chambers, Lin; Holzer, Missy; Moore, Susan

    2009-01-01

    My NASA Data (Chambers et al. 2008) is a teaching tool available on NASA's website that offers microsets of real data in an easily accessible, user-friendly format. In this article, the authors describe a lesson plan based on an activity from My NASA Data, in which students explore parts of the United States where they would want to live if they…

  6. Harnessing science to improve safety.

    Science.gov (United States)

    Baillie, Jonathan

    2011-01-01

    Examining the effectiveness of various wet surface cleaning methods in combating harmful microorganisms in a hospital ward, understanding different healthcare cleaning regimes' impact on reducing slips and trips, evaluating the protection offered by surgical masks against influenza bioaerosols, and independently testing tower crane safety following a number of fatal incidents, are among the broad spectrum of recent projects undertaken by the Buxton-headquartered Health and Safety Laboratory (HSL). As HEJ editor Jonathan Baillie discovered from the organisation's healthcare and patient safety lead, Darren Whitehouse, with around 350 scientists skilled in everything from microbiology to occupational psychology, the range of scientific guidance, expertise, advice, testing, training, and investigation, that the HSL can offer to the healthcare sector is perhaps unrivalled throughout Europe.

  7. harnessing_ai_20180319.pptx

    OpenAIRE

    Johnson, Matthew

    2018-01-01

    There have been great strides made in Artificial Intelligence in recent years, but it is often not clear how to apply these recent advances to new problems. In this talk I propose one method of thinking about problems in the sciences and how to find the right AI techniques to aid in research.

  8. HARNESSING INTELLECTUAL PROPERTY FOR DEVELOPMENT ...

    African Journals Online (AJOL)

    10332324

    @uct.ac.za. This article is partially derived from the author‟s thesis: CB Ncube Intellectual Property Protection for e-Commerce Business Methods in South Africa: Envisioning an Equitable Model for SMEs in the. Tourism Industry (PhD thesis ...

  9. Vi har sunget, vi har grædt, vi har grint

    DEFF Research Database (Denmark)

    Thurah, Thomas

    Fra omslaget: Benny Andersen og Povl Dissing fortæller her for første gang om deres lange liv med musikken, fra barndommens øvelser med klaver, trommer og trompet hjemme i dagligstuen til samarbejdet med symforniorkestre på vores største scener....

  10. Telomerase expression extends the proliferative life-span and maintains the osteogenic potential of human bone marrow stromal cells

    DEFF Research Database (Denmark)

    Simonsen, Janne Lytoft; Rosada, Cecilia; Serakinci, Nedime

    2002-01-01

    . The transduced cells have now undergone more than 260 population doublings (PD) and continue to proliferate, whereas control cells underwent senescence-associated proliferation arrest after 26 PD. The cells maintained production of osteoblastic markers and differentiation potential during continuous subculturing......, did not form tumors, and had a normal karyotype. When implanted subcutaneously in immunodeficient mice, the transduced cells formed more bone than did normal cells. These results suggest that ectopic expression of telomerase in hMSCs prevents senescence-associated impairment of osteoblast functions.......Human bone marrow stromal cells (hMSCs) were stably transduced by a retroviral vector containing the gene for the catalytic subunit of human telomerase (hTERT). Transduced cells (hMSC-TERTs) had telomerase activity, and the mean telomere length was increased as compared with that of control cells...

  11. The effects of 5-fluorouracil and doxorubicin on expression of human immunodeficiency virus type 1 long terminal repeat

    International Nuclear Information System (INIS)

    Panozzo, J.; Akan, E.; Griffiths, T.D.

    1996-01-01

    Previous work by many groups has documented induction of the HIV-LTR following exposure of cells to ultraviolet light and other DNA damaging agents. Our experiments set out to determine the relative activation or repression of the HIV-LTR in response to two classes of chemotherapeutic agents: Doxorubicin is a DNA-damage inducing agent, and 5-fluorouracil has an antimetabolic mode of action. Using HeLa cells stably transfected with a construct in which HIV-LTR drives expression of the chloramphenicol acetyl transferase reporter gene, we demonstrated an up to 10-fold induction following doxorubicin treatment in 24 h post-treatment. This induction was repressed by treatment with salicylic acid, suggesting a role for prostaglandin/cyclo-oxygenase pathways and/or NFKB in the inductive response. Induction by 5-fluorouracil, in contrast, was more modest (two-fold at most) though it was consistently elevated over controls

  12. Purification and characterization of recombinant full-length and protease domain of murine MMP-9 expressed in Drosophila S2 cells

    DEFF Research Database (Denmark)

    Rasch, Morten G; Lund, Ida K; Illemann, Martin

    2010-01-01

    Matrix metalloproteinase-9 (MMP-9) is a 92-kDa soluble pro-enzyme implicated in pathological events including cancer invasion. It is therefore an attractive target for therapeutic intervention studies in mouse models. Development of inhibitors requires sufficient amounts of correctly folded murine...... MMP-9. Constructs encoding zymogens of full-length murine MMP-9 and a version lacking the O-glycosylated linker region and hemopexin domains were therefore generated and expressed in stably transfected Drosophila S2 insect cells. After 7 days of induction the expression levels of the full....... No immunoreactivity was observed when the antibody was probed against skin wound material from MMP-9 deficient mice. In conclusion, we have generated and purified two proteolytically active recombinant murine MMP-9 protein constructs, which are critical reagents for future cancer drug discovery studies....

  13. Purification and characterization of recombinant full-length and protease domain of murine MMP-9 expressed in Drosophila S2 cells

    DEFF Research Database (Denmark)

    Rasch, Morten G; Lund, Ida K; Illemann, Martin

    2010-01-01

    MMP-9. Constructs encoding zymogens of full-length murine MMP-9 and a version lacking the O-glycosylated linker region and hemopexin domains were therefore generated and expressed in stably transfected Drosophila S2 insect cells. After 7 days of induction the expression levels of the full......-length and truncated versions were 5 mg/l and 2 mg/l, respectively. The products were >95% pure after gelatin Sepharose chromatography and possessed proteolytic activity when analyzed by gelatin zymography. Using the purified full-length murine MMP-9 we raised polyclonal antibodies by immunizations of rabbits......Matrix metalloproteinase-9 (MMP-9) is a 92-kDa soluble pro-enzyme implicated in pathological events including cancer invasion. It is therefore an attractive target for therapeutic intervention studies in mouse models. Development of inhibitors requires sufficient amounts of correctly folded murine...

  14. Impact of the Ku complex on HIV-1 expression and latency.

    Directory of Open Access Journals (Sweden)

    Gwenola Manic

    Full Text Available Ku, a cellular complex required for human cell survival and involved in double strand break DNA repair and multiple other cellular processes, may modulate retroviral multiplication, although the precise mechanism through which it acts is still controversial. Recently, Ku was identified as a possible anti-human immunodeficiency virus type 1 (HIV-1 target in human cells, in two global approaches. Here we investigated the role of Ku on the HIV-1 replication cycle by analyzing the expression level of a panel of non-replicative lentiviral vectors expressing the green fluorescent protein in human colorectal carcinoma HCT 116 cells, stably or transiently depleted of Ku. We found that in this cellular model the depletion of Ku did not affect the efficiency of (pre-integrative steps but decreased the early HIV-1 expression by acting at the transcriptional level. This negative effect was specific of the HIV-1 promoter, required the obligatory step of viral DNA integration and was reversed by transient depletion of p53. We also provided evidence on a direct binding of Ku to HIV-1 LTR in transduced cells. Ku not only promotes the early transcription from the HIV-1 promoter, but also limits the constitution of viral latency. Moreover, in the presence of a normal level of Ku, HIV-1 expression was gradually lost over time, likely due to the counter-selection of HIV-1-expressing cells. On the contrary, the reactivation of transgene expression from HIV-1 by means of trichostatin A- or tumor necrosis factor α-administration was enhanced under condition of Ku haplodepletion, suggesting a phenomenon of provirus latency. These observations plead in favor of the hypothesis that Ku has an impact on HIV-1 expression and latency at early- and mid-time after integration.

  15. Active RNA replication of hepatitis C virus downregulates CD81 expression.

    Directory of Open Access Journals (Sweden)

    Po-Yuan Ke

    Full Text Available So far how hepatitis C virus (HCV replication modulates subsequent virus growth and propagation still remains largely unknown. Here we determine the impact of HCV replication status on the consequential virus growth by comparing normal and high levels of HCV RNA expression. We first engineered a full-length, HCV genotype 2a JFH1 genome containing a blasticidin-resistant cassette inserted at amino acid residue of 420 in nonstructural (NS protein 5A, which allowed selection of human hepatoma Huh7 cells stably-expressing HCV. Short-term establishment of HCV stable cells attained a highly-replicating status, judged by higher expressions of viral RNA and protein as well as higher titer of viral infectivity as opposed to cells harboring the same genome without selection. Interestingly, maintenance of highly-replicating HCV stable cells led to decreased susceptibility to HCV pseudotyped particle (HCVpp infection and downregulated cell surface level of CD81, a critical HCV entry (coreceptor. The decreased CD81 cell surface expression occurred through reduced total expression and cytoplasmic retention of CD81 within an endoplasmic reticulum -associated compartment. Moreover, productive viral RNA replication in cells harboring a JFH1 subgenomic replicon containing a similar blasticidin resistance gene cassette in NS5A and in cells robustly replicating full-length infectious genome also reduced permissiveness to HCVpp infection through decreasing the surface expression of CD81. The downregulation of CD81 surface level in HCV RNA highly-replicating cells thus interfered with reinfection and led to attenuated viral amplification. These findings together indicate that the HCV RNA replication status plays a crucial determinant in HCV growth by modulating the expression and intracellular localization of CD81.

  16. Using RNA-seq data to select reference genes for normalizing gene expression in apple roots.

    Directory of Open Access Journals (Sweden)

    Zhe Zhou

    Full Text Available Gene expression in apple roots in response to various stress conditions is a less-explored research subject. Reliable reference genes for normalizing quantitative gene expression data have not been carefully investigated. In this study, the suitability of a set of 15 apple genes were evaluated for their potential use as reliable reference genes. These genes were selected based on their low variance of gene expression in apple root tissues from a recent RNA-seq data set, and a few previously reported apple reference genes for other tissue types. Four methods, Delta Ct, geNorm, NormFinder and BestKeeper, were used to evaluate their stability in apple root tissues of various genotypes and under different experimental conditions. A small panel of stably expressed genes, MDP0000095375, MDP0000147424, MDP0000233640, MDP0000326399 and MDP0000173025 were recommended for normalizing quantitative gene expression data in apple roots under various abiotic or biotic stresses. When the most stable and least stable reference genes were used for data normalization, significant differences were observed on the expression patterns of two target genes, MdLecRLK5 (MDP0000228426, a gene encoding a lectin receptor like kinase and MdMAPK3 (MDP0000187103, a gene encoding a mitogen-activated protein kinase. Our data also indicated that for those carefully validated reference genes, a single reference gene is sufficient for reliable normalization of the quantitative gene expression. Depending on the experimental conditions, the most suitable reference genes can be specific to the sample of interest for more reliable RT-qPCR data normalization.

  17. Widespread expression of SAA and Hp RNA in bovine tissues after evaluation of suitable reference genes.

    Science.gov (United States)

    Lecchi, Cristina; Dilda, Francesca; Sartorelli, Paola; Ceciliani, Fabrizio

    2012-01-15

    The serum amyloid A (SAA) and haptoglobin (Hp) are the most prominent acute phase proteins (APPs) in cow. Liver mainly produces APPs, but extra hepatic expression has also been demonstrated in some tissues. The major aim of the present study was to assess the constitutive SAA and Hp mRNA expression by quantitative PCR (qPCR) in a wide panel of 33 bovine tissues, including gastrointestinal tract, respiratory system, urogenital system, mammary gland, hematopoietic system, central nervous system, eye, thyroid and heart. Normalization of gene expression in different samples requires reference genes, which are stably expressed. Therefore, seven reference genes were investigated (ACTB, GAPDH, HMBS, SDHA, YWHAZ, SF3A1, EEF1A2) and three genes, namely SF3A1, HMBS and ACTB, were selected after assessing their stability with geNorm™ and NormFinder(©) softwares. The qPCR analysis confirmed liver as the principal source of SAA and Hp, but also identified both APPs' mRNA in almost all tissues. The highest expression rate of SAA was found in thyroid, followed by pancreas and submandibulary gland. Hp mRNA expression was detected at high concentration in pancreas and submandibulary gland. The present data indicated a widespread expression of SAA and Hp also in non pathological conditions, thus envisaging a possible role as immunomodulatory and protective molecules. To understand where SAA and Hp come from is the prerequisite to their utilization as Acute Phase Reaction biomarkers. Copyright © 2011 Elsevier B.V. All rights reserved.

  18. Reference genes for gene expression studies in wheat flag leaves grown under different farming conditions

    Directory of Open Access Journals (Sweden)

    Cordeiro Raposo Fernando

    2011-09-01

    Full Text Available Abstract Background Internal control genes with highly uniform expression throughout the experimental conditions are required for accurate gene expression analysis as no universal reference genes exists. In this study, the expression stability of 24 candidate genes from Triticum aestivum cv. Cubus flag leaves grown under organic and conventional farming systems was evaluated in two locations in order to select suitable genes that can be used for normalization of real-time quantitative reverse-transcription PCR (RT-qPCR reactions. The genes were selected among the most common used reference genes as well as genes encoding proteins involved in several metabolic pathways. Findings Individual genes displayed different expression rates across all samples assayed. Applying geNorm, a set of three potential reference genes were suitable for normalization of RT-qPCR reactions in winter wheat flag leaves cv. Cubus: TaFNRII (ferredoxin-NADP(H oxidoreductase; AJ457980.1, ACT2 (actin 2; TC234027, and rrn26 (a putative homologue to RNA 26S gene; AL827977.1. In addition of these three genes that were also top-ranked by NormFinder, two extra genes: CYP18-2 (Cyclophilin A, AY456122.1 and TaWIN1 (14-3-3 like protein, AB042193 were most consistently stably expressed. Furthermore, we showed that TaFNRII, ACT2, and CYP18-2 are suitable for gene expression normalization in other two winter wheat varieties (Tommi and Centenaire grown under three treatments (organic, conventional and no nitrogen and a different environment than the one tested with cv. Cubus. Conclusions This study provides a new set of reference genes which should improve the accuracy of gene expression analyses when using wheat flag leaves as those related to the improvement of nitrogen use efficiency for cereal production.

  19. Resveratrol inhibits Cdk5 activity through regulation of p35 expression

    Directory of Open Access Journals (Sweden)

    Kulkarni Ashok B

    2011-07-01

    Full Text Available Abstract Background We have previously reported that cyclin-dependent kinase 5 (Cdk5 participates in the regulation of nociceptive signaling. Through activation of the ERK1/2 pathway, Tumor Necrosis Factor-α (TNF-α induces expression of Egr-1. This results in the sustained and robust expression of p35, a coactivator of Cdk5, in PC12 cells, thereby increasing Cdk5 kinase activity. The aim of our present study was to test whether resveratrol, a polyphenolic compound with known analgesic activity, can regulate Cdk5/p35 activity. Results Here we used a cell-based assay in which a p35 promoter-luciferase construct was stably transfected in PC12 cells. Our studies demonstrate that resveratrol inhibits p35 promoter activity and also blocks the TNF-α mediated increase in Cdk5 activity in PC12 cells. Resveratrol also inhibits p35 expression and blocks the TNF-α mediated increase in Cdk5 activity in DRG neurons. In the presence of resveratrol, the MEK inhibitor decreased p35 promoter activity, whereas the inhibitors of p38 MAPK, JNK and NF-κB increased p35 promoter activity, indicating that these pathways regulate p35 expression differently. The TNF-α-mediated increase in Egr-1 expression was decreased by resveratrol treatment with a concomitant reduction in p35 expression and protein levels, resulting in reduced Cdk5 kinase activity. Conclusions We demonstrate here that resveratrol regulates p35 promoter activity in PC12 cells and DRG neurons. Most importantly, resveratrol blocks the TNF-α-mediated increase in p35 promoter activity, thereby reducing p35 expression and subsequent Cdk5 kinase activity. This new molecular mechanism adds to the known analgesic effects of resveratrol and confirms the need for identifying new analgesics based on their ability to inhibit Cdk5 activity for effective treatment of pain.

  20. Evaluation of suitable reference genes for gene expression studies in bovine muscular tissue

    Directory of Open Access Journals (Sweden)

    Dunner Susana

    2008-09-01

    Full Text Available Abstract Background Real-time reverse transcriptase quantitative polymerase chain reaction (real-time RTqPCR is a technique used to measure mRNA species copy number as a way to determine key genes involved in different biological processes. However, the expression level of these key genes may vary among tissues or cells not only as a consequence of differential expression but also due to different factors, including choice of reference genes to normalize the expression levels of the target genes; thus the selection of reference genes is critical for expression studies. For this purpose, ten candidate reference genes were investigated in bovine muscular tissue. Results The value of stability of ten candidate reference genes included in three groups was estimated: the so called 'classical housekeeping' genes (18S, GAPDH and ACTB, a second set of genes used in expression studies conducted on other tissues (B2M, RPII, UBC and HMBS and a third set of novel genes (SF3A1, EEF1A2 and CASC3. Three different statistical algorithms were used to rank the genes by their stability measures as produced by geNorm, NormFinder and Bestkeeper. The three methods tend to agree on the most stably expressed genes and the least in muscular tissue. EEF1A2 and HMBS followed by SF3A1, ACTB, and CASC3 can be considered as stable reference genes, and B2M, RPII, UBC and GAPDH would not be appropriate. Although the rRNA-18S stability measure seems to be within the range of acceptance, its use is not recommended because its synthesis regulation is not representative of mRNA levels. Conclusion Based on geNorm algorithm, we propose the use of three genes SF3A1, EEF1A2 and HMBS as references for normalization of real-time RTqPCR in muscle expression studies.