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Sample records for a-kinase anchoring protein

  1. A systematic evaluation of protein kinase A-A-kinase anchoring protein interaction motifs

    NARCIS (Netherlands)

    Burgers, Pepijn P; van der Heyden, MAG; Kok, Bart; Heck, Albert J R; Scholten, Arjen

    2015-01-01

    Protein kinase A (PKA) in vertebrates is localized to specific locations in the cell via A-kinase anchoring proteins (AKAPs). The regulatory subunits of the four PKA isoforms (RIα, RIβ, RIIα, and RIIβ) each form a homodimer, and their dimerization domain interacts with a small helical region present

  2. A kinase-anchoring proteins and adenylyl cyclase in cardiovascular physiology and pathology.

    Science.gov (United States)

    Efendiev, Riad; Dessauer, Carmen W

    2011-10-01

    3'-5'-Cyclic adenosine monophosphate (cAMP), generated by adenylyl cyclase (AC), serves as a second messenger in signaling pathways regulating many aspects of cardiac physiology, including contraction rate and action potential duration, and in the pathophysiology of hypertrophy and heart failure. A kinase-anchoring proteins localize the effect of cAMP in space and time by organizing receptors, AC, protein kinase A, and other components of the cAMP cascade into multiprotein complexes. In this review, we discuss how the interaction of A kinase-anchoring proteins with distinct AC isoforms affects cardiovascular physiology.

  3. A-kinase anchoring proteins contribute to loss of E-cadherin and bronchial epithelial barrier by cigarette smoke

    NARCIS (Netherlands)

    Oldenburger, Anouk; Poppinga, Wilfred J; Kos, Fleur; de Bruin, Harold G; Rijks, Wolter F; Heijink, Hilde; Timens, Wim; Meurs, Hermanus; Maarsingh, Harm; Schmidt, Martina

    2014-01-01

    Airway epithelium, which forms the first barrier towards environmental insults, is disturbed by cigarette smoking, a major risk factor for developing chronic obstructive pulmonary disease (COPD). A-kinase anchoring proteins (AKAP) maintain endothelial barrier function and coordinate subcellular loca

  4. Promotion and inhibition of cardiac hypertrophy by A-kinase anchor proteins.

    Science.gov (United States)

    Blant, Alexandra; Czubryt, Michael P

    2012-09-01

    Originally identified as mediators of cyclic adenosine monophosphate (cAMP) and protein kinase A signaling, A-kinase anchor proteins (AKAPs) are now recognized as a diverse family of molecular scaffolds capable of interacting with many other proteins. Members of the AKAP family within the heart can take on either pro- or anti-hypertrophic roles by interacting with a myriad of protein kinases and phosphatases in the process. AKAPs often form the core of large signaling complexes (or signalosomes) that allow multiple pathways to converge and functionally intertwine. Approximately 30% of AKAPs discovered to date are expressed in the heart, but the functions of many of these remain to be discovered. This review focuses on AKAPs that have been demonstrated to play roles in mediating cardiac hypertrophy.

  5. Molecular evolution of a-kinase anchoring protein (AKAP-7: implications in comparative PKA compartmentalization

    Directory of Open Access Journals (Sweden)

    Johnson Keven R

    2012-07-01

    Full Text Available Abstract Background A-Kinase Anchoring Proteins (AKAPs are molecular scaffolding proteins mediating the assembly of multi-protein complexes containing cAMP-dependent protein kinase A (PKA, directing the kinase in discrete subcellular locations. Splice variants from the AKAP7 gene (AKAP15/18 are vital components of neuronal and cardiac phosphatase complexes, ion channels, cardiac Ca2+ handling and renal water transport. Results Shown in evolutionary analyses, the formation of the AKAP7-RI/RII binding domain (required for AKAP/PKA-R interaction corresponds to vertebrate-specific gene duplication events in the PKA-RI/RII subunits. Species analyses of AKAP7 splice variants shows the ancestral AKAP7 splice variant is AKAP7α, while the ancestral long form AKAP7 splice variant is AKAP7γ. Multi-species AKAP7 gene alignments, show the recent formation of AKAP7δ occurs with the loss of native AKAP7γ in rats and basal primates. AKAP7 gene alignments and two dimensional Western analyses indicate that AKAP7γ is produced from an internal translation-start site that is present in the AKAP7δ cDNA of mice and humans but absent in rats. Immunofluorescence analysis of AKAP7 protein localization in both rat and mouse heart suggests AKAP7γ replaces AKAP7δ at the cardiac sarcoplasmic reticulum in species other than rat. DNA sequencing identified Human AKAP7δ insertion-deletions (indels that promote the production of AKAP7γ instead of AKAP7δ. Conclusions This AKAP7 molecular evolution study shows that these vital scaffolding proteins developed in ancestral vertebrates and that independent mutations in the AKAP7 genes of rodents and early primates has resulted in the recent formation of AKAP7δ, a splice variant of likely lesser importance in humans than currently described.

  6. Muscle A-Kinase Anchoring Protein-α is an Injury-Specific Signaling Scaffold Required for Neurotrophic- and Cyclic Adenosine Monophosphate-Mediated Survival

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    Yan Wang

    2015-12-01

    Full Text Available Neurotrophic factor and cAMP-dependent signaling promote the survival and neurite outgrowth of retinal ganglion cells (RGCs after injury. However, the mechanisms conferring neuroprotection and neuroregeneration downstream to these signals are unclear. We now reveal that the scaffold protein muscle A-kinase anchoring protein-α (mAKAPα is required for the survival and axon growth of cultured primary RGCs. Although genetic deletion of mAKAPα early in prenatal RGC development did not affect RGC survival into adulthood, nor promoted the death of RGCs in the uninjured adult retina, loss of mAKAPα in the adult increased RGC death after optic nerve crush. Importantly, mAKAPα was required for the neuroprotective effects of brain-derived neurotrophic factor and cyclic adenosine-monophosphate (cAMP after injury. These results identify mAKAPα as a scaffold for signaling in the stressed neuron that is required for RGC neuroprotection after optic nerve injury.

  7. The Scaffold Protein Muscle A-Kinase Anchoring Protein β Orchestrates Cardiac Myocyte Hypertrophic Signaling Required for the Development of Heart Failure

    Science.gov (United States)

    Kritzer, Michael D.; Li, Jinliang; Passariello, Catherine L.; Gayanilo, Marjorie; Thakur, Hrishikesh; Dayan, Joseph; Dodge-Kafka, Kimberly; Kapiloff, Michael S.

    2014-01-01

    Background Cardiac myocyte hypertrophy is regulated by an extensive intracellular signal transduction network. In vitro evidence suggests that the scaffold protein muscle A-kinase anchoring protein β (mAKAPβ) serves as a nodal organizer of hypertrophic signaling. However, the relevance of mAKAPβ signalosomes to pathological remodeling and heart failure in vivo remains unknown. Methods and Results Using conditional, cardiac myocyte–specific gene deletion, we now demonstrate that mAKAPβ expression in mice is important for the cardiac hypertrophy induced by pressure overload and catecholamine toxicity. mAKAPβ targeting prevented the development of heart failure associated with long-term transverse aortic constriction, conferring a survival benefit. In contrast to 29% of control mice (n=24), only 6% of mAKAPβ knockout mice (n=31) died in the 16 weeks of pressure overload (P=0.02). Accordingly, mAKAPβ knockout inhibited myocardial apoptosis and the development of interstitial fibrosis, left atrial hypertrophy, and pulmonary edema. This improvement in cardiac status correlated with the attenuated activation of signaling pathways coordinated by the mAKAPβ scaffold, including the decreased phosphorylation of protein kinase D1 and histone deacetylase 4 that we reveal to participate in a new mAKAP signaling module. Furthermore, mAKAPβ knockout inhibited pathological gene expression directed by myocyte-enhancer factor-2 and nuclear factor of activated T-cell transcription factors that associate with the scaffold. Conclusions mAKAPβ orchestrates signaling that regulates pathological cardiac remodeling in mice. Targeting of the underlying physical architecture of signaling networks, including mAKAPβ signalosome formation, may constitute an effective therapeutic strategy for the prevention and treatment of pathological remodeling and heart failure. PMID:24812305

  8. A Kinase Anchoring Protein 9 Is a Novel Myosin VI Binding Partner That Links Myosin VI with the PKA Pathway in Myogenic Cells

    Directory of Open Access Journals (Sweden)

    Justyna Karolczak

    2015-01-01

    Full Text Available Myosin VI (MVI is a unique motor protein moving towards the minus end of actin filaments unlike other known myosins. Its important role has recently been postulated for striated muscle and myogenic cells. Since MVI functions through interactions of C-terminal globular tail (GT domain with tissue specific partners, we performed a search for MVI partners in myoblasts and myotubes using affinity chromatography with GST-tagged MVI-GT domain as a bait. A kinase anchoring protein 9 (AKAP9, a regulator of PKA activity, was identified by means of mass spectrometry as a possible MVI interacting partner both in undifferentiated and differentiating myoblasts and in myotubes. Coimmunoprecipitation and proximity ligation assay confirmed that both proteins could interact. MVI and AKAP9 colocalized at Rab5 containing early endosomes. Similarly to MVI, the amount of AKAP9 decreased during myoblast differentiation. However, in MVI-depleted cells, both cAMP and PKA levels were increased and a change in the MVI motor-dependent AKAP9 distribution was observed. Moreover, we found that PKA phosphorylated MVI-GT domain, thus implying functional relevance of MVI-AKAP9 interaction. We postulate that this novel interaction linking MVI with the PKA pathway could be important for targeting AKAP9-PKA complex within cells and/or providing PKA to phosphorylate MVI tail domain.

  9. A Kinase Anchoring Protein 9 Is a Novel Myosin VI Binding Partner That Links Myosin VI with the PKA Pathway in Myogenic Cells.

    Science.gov (United States)

    Karolczak, Justyna; Sobczak, Magdalena; Skowronek, Krzysztof; Rędowicz, Maria Jolanta

    2015-01-01

    Myosin VI (MVI) is a unique motor protein moving towards the minus end of actin filaments unlike other known myosins. Its important role has recently been postulated for striated muscle and myogenic cells. Since MVI functions through interactions of C-terminal globular tail (GT) domain with tissue specific partners, we performed a search for MVI partners in myoblasts and myotubes using affinity chromatography with GST-tagged MVI-GT domain as a bait. A kinase anchoring protein 9 (AKAP9), a regulator of PKA activity, was identified by means of mass spectrometry as a possible MVI interacting partner both in undifferentiated and differentiating myoblasts and in myotubes. Coimmunoprecipitation and proximity ligation assay confirmed that both proteins could interact. MVI and AKAP9 colocalized at Rab5 containing early endosomes. Similarly to MVI, the amount of AKAP9 decreased during myoblast differentiation. However, in MVI-depleted cells, both cAMP and PKA levels were increased and a change in the MVI motor-dependent AKAP9 distribution was observed. Moreover, we found that PKA phosphorylated MVI-GT domain, thus implying functional relevance of MVI-AKAP9 interaction. We postulate that this novel interaction linking MVI with the PKA pathway could be important for targeting AKAP9-PKA complex within cells and/or providing PKA to phosphorylate MVI tail domain.

  10. How anchoring proteins shape pain.

    Science.gov (United States)

    Fischer, Michael J M; McNaughton, Peter A

    2014-09-01

    Cellular responsiveness to external stimuli can be altered by extracellular mediators which activate membrane receptors, in turn signalling to the intracellular space via calcium, cyclic nucleotides, membrane lipids or enzyme activity. These signalling events trigger a cascade leading to an effector which can be a channel, an enzyme or a transcription factor. The effectiveness of these intracellular events is enhanced when they are maintained in close proximity by anchoring proteins, which assemble complexes of signalling molecules such as kinases together with their targets, and in this way enhance both the speed and the precision of intracellular signalling. The A kinase anchoring protein (AKAP) family are adaptor proteins originally named for their ability to associate Protein Kinase A and its targets, but several other enzymes bound by AKAPs have now been found and a wide variety of target structures has been described. This review provides an overview of anchoring proteins involved in pain signalling. The key anchoring proteins and their ion channel targets in primary sensory neurons responding to painful stimuli (nociceptors) are discussed.

  11. Anchored design of protein-protein interfaces.

    Directory of Open Access Journals (Sweden)

    Steven M Lewis

    Full Text Available BACKGROUND: Few existing protein-protein interface design methods allow for extensive backbone rearrangements during the design process. There is also a dichotomy between redesign methods, which take advantage of the native interface, and de novo methods, which produce novel binders. METHODOLOGY: Here, we propose a new method for designing novel protein reagents that combines advantages of redesign and de novo methods and allows for extensive backbone motion. This method requires a bound structure of a target and one of its natural binding partners. A key interaction in this interface, the anchor, is computationally grafted out of the partner and into a surface loop on the design scaffold. The design scaffold's surface is then redesigned with backbone flexibility to create a new binding partner for the target. Careful choice of a scaffold will bring experimentally desirable characteristics into the new complex. The use of an anchor both expedites the design process and ensures that binding proceeds against a known location on the target. The use of surface loops on the scaffold allows for flexible-backbone redesign to properly search conformational space. CONCLUSIONS AND SIGNIFICANCE: This protocol was implemented within the Rosetta3 software suite. To demonstrate and evaluate this protocol, we have developed a benchmarking set of structures from the PDB with loop-mediated interfaces. This protocol can recover the correct loop-mediated interface in 15 out of 16 tested structures, using only a single residue as an anchor.

  12. Anchoring of proteins to lactic acid bacteria

    NARCIS (Netherlands)

    Leenhouts, K; Buist, Girbe; Kok, Jan

    1999-01-01

    The anchoring of proteins to the cell surface of lactic acid bacteria (LAB) using genetic techniques is an exciting and emerging research area that holds great promise for a wide variety of biotechnological applications. This paper reviews five different types of anchoring domains that have been exp

  13. Biomedical applications of glycosylphosphatidylinositol-anchored proteins

    Science.gov (United States)

    Heider, Susanne; Dangerfield, John A.

    2016-01-01

    Glycosylphosphatidylinositol (GPI)-anchored proteins (GPI-APs) use a unique posttranslational modification to link proteins to lipid bilayer membranes. The anchoring structure consists of both a lipid and carbohydrate portion and is highly conserved in eukaryotic organisms regarding its basic characteristics, yet highly variable in its molecular details. The strong membrane targeting property has made the anchors an interesting tool for biotechnological modification of lipid membrane-covered entities from cells through extracellular vesicles to enveloped virus particles. In this review, we will take a closer look at the mechanisms and fields of application for GPI-APs in lipid bilayer membrane engineering and discuss their advantages and disadvantages for biomedicine. PMID:27542385

  14. Endocytosis of glycosylphosphatidylinositol-anchored proteins

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    Sabharanjak Shefali

    2009-10-01

    Full Text Available Abstract Glycosylphosphatidylinositol-anchored proteins (GPI-APs represent an interesting amalgamation of the three basic kinds of cellular macromolecules viz. proteins, carbohydrates and lipids. An unusually hybrid moiety, the GPI-anchor is expressed in a diverse range of organisms from parasites to mammalian cells and serves to anchor a large number of functionally diverse proteins and has been the center of attention in scientific debate for some time now. Membrane organization of GPI-APs into laterally-organized cholesterol-sphingolipid ordered membrane domains or "rafts" and endocytosis of GPI-APs has been intensely debated. Inclusion into or exclusion from these membrane domains seems to be the critical factor in determining the endocytic mechanisms and intracellular destinations of GPI-APs. The intracellular signaling as well as endocytic trafficking of GPI-APs is critically dependent upon the cell surface organization of GPI-APs, and the associations with these lipid rafts play a vital role during these processes. The mechanism of endocytosis for GPI-APs may differ from other cellular endocytic pathways, such as those mediated by clathrin-coated pits (caveolae, and is necessary for unique biological functions. Numerous intracellular factors are involved in and regulate the endocytosis of GPI-APs, and these may be variably dependent on cell-type. The central focus of this article is to describe the significance of the endocytosis of GPI-APs on a multitude of biological processes, ranging from nutrient-uptake to more complex immune responses. Ultimately, a thorough elucidation of GPI-AP mediated signaling pathways and their regulatory elements will enhance our understanding of essential biological processes and benefit as components of disease intervention strategies.

  15. FragAnchor: A Large-Scale Predictor of Glycosylphosphatidylinositol Anchors in Eukaryote Protein Sequences by Qualitative Scoring

    Institute of Scientific and Technical Information of China (English)

    2007-01-01

    A glycosylphosphatidylinositol (GPI) anchor is a common but complex C-terminal post-translational modification of extracellular proteins in eukaryotes. Here we investigate the problem of correctly annotating GPI-anchored proteins for the growing number of sequences in public databases. We developed a computational system, called FragAnchor, based on the tandem use of a neural network (NN) and a hidden Markov model (HMM). Firstly, NN selects potential GPI-anchored proteins in a dataset, then HMM parses these potential GPI signals and refines the prediction by qualitative scoring. FragAnchor correctly predicted 91% of all the GPI-anchored proteins annotated in the Swiss-Prot database.In a large-scale analysis of 29 eukaryote proteomes, FragAnchor predicted that the percentage of highly probable GPI-anchored proteins is between 0.21% and 2.01%. The distinctive feature of FragAnchor, compared with other systems,is that it targets only the C-terminus of a protein, making it less sensitive to the background noise found in databases and possible incomplete protein sequences. Moreover, FragAnchor can be used to predict GPI-anchored proteins in all eukaryotes. Finally, by using qualitative scoring, the predictions combine both sensitivity and information content. The predictor is publicly available at http: // navet. ics. hawaii.edu/~fraganchor/NNHMM/NNHMM.html.

  16. Conditional depletion of nuclear proteins by the Anchor Away system.

    Science.gov (United States)

    Fan, Xiaochun; Geisberg, Joseph V; Wong, Koon Ho; Jin, Yi

    2011-01-01

    Nuclear proteins play key roles in the regulation of many important cellular processes. In Saccharomyces cerevisiae, many genes encoding nuclear proteins are essential. This unit describes a method termed Anchor Away that can be used to conditionally and rapidly deplete nuclear proteins of interest. It involves conditional export of the protein of interest out of the nucleus and its subsequent sequestration in the cytoplasm. This method can be used to simultaneously deplete multiple proteins from the nucleus.

  17. Proteomic analysis of glycosylphosphatidylinositol-anchored membrane proteins

    DEFF Research Database (Denmark)

    Elortza, Felix; Nühse, Thomas S; Foster, Leonard J

    2003-01-01

    Glycosylphosphatidylinositol-anchored proteins (GPI-APs) are a functionally and structurally diverse family of post-translationally modified membrane proteins found mostly in the outer leaflet of the plasma membrane in a variety of eukaryotic cells. Although the general role of GPI-APs remains un...

  18. Proteomic analysis of GPI-anchored membrane proteins

    DEFF Research Database (Denmark)

    Jung, Hye Ryung; Jensen, Ole Nørregaard

    2006-01-01

    Glycosyl-phosphatidyl-inositol-anchored proteins (GPI-APs) represent a subset of post-translationally modified proteins that are tethered to the outer leaflet of the plasma membrane via a C-terminal GPI anchor. GPI-APs are found in a variety of eukaryote species, from pathogenic microorganisms...... to humans. GPI-APs confer important cellular functions as receptors, enzymes and scaffolding molecules. Specific enzymes and detergent extraction methods combined with separation technologies and mass spectrometry permit proteomic analysis of GPI-APs from plasma membrane preparations to reveal cell...

  19. Tail-anchored membrane proteins: exploring the complex diversity of tail-anchored-protein targeting in plant cells.

    Science.gov (United States)

    Abell, Ben M; Mullen, Robert T

    2011-02-01

    Tail-anchored (TA) proteins are special class of integral membrane proteins that in recent years have received a considerable amount of attention due to their diverse cellular functions and unique targeting and insertion mechanisms. Defined by the presence of a single, hydrophobic membrane-spanning domain at or near their C terminus, TA proteins must be inserted into membranes post-translationally and are orientated such that their larger N-terminal domain (most often the functional domain) faces the cytosol, while their shorter C-terminal domain faces the interior of the organelle. The C-terminal domain of TA proteins also usually contains the information responsible for their selective targeting to the proper subcellular membrane, a process that, based primarily on studies with yeasts and mammals, appears to be highly complex due to the presence of multiple pathways. Within this context, we discuss here the biogenesis of plant TA proteins and the potential for hundreds of new TA proteins identified via bioinformatics screens to contribute to the already remarkable number of roles that this class of membrane proteins participates in throughout plant growth and development.

  20. The association between glycosylphosphatidylinositol-anchored proteins and heterotrimeric G protein alpha subunits in lymphocytes.

    OpenAIRE

    1996-01-01

    Glycosylphosphatidylinositol (GPI)-anchored proteins are nonmembrane spanning cell surface proteins that have been demonstrated to be signal transduction molecules. Because these proteins do not extend into the cytoplasm, the mechanism by which cross-linking of these molecules leads to intracellular signal transduction events is obscure. Previous analysis has indicated that these proteins are associated with src family member tyrosine kinases; however, the role this interaction plays in the g...

  1. Solid-state NMR spectra of lipid-anchored proteins under magic angle spinning.

    Science.gov (United States)

    Nomura, Kaoru; Harada, Erisa; Sugase, Kenji; Shimamoto, Keiko

    2014-03-01

    Solid-state NMR is a promising tool for elucidating membrane-related biological phenomena. We achieved the measurement of high-resolution solid-state NMR spectra for a lipid-anchored protein embedded in lipid bilayers under magic angle spinning (MAS). To date, solid-state NMR measurements of lipid-anchored proteins have not been accomplished due to the difficulty in supplying sufficient amount of stable isotope labeled samples in the overexpression of lipid-anchored proteins requiring complex posttranslational modification. We designed a pseudo lipid-anchored protein in which the protein component was expressed in E. coli and attached to a chemically synthesized lipid-anchor mimic. Using two types of membranes, liposomes and bicelles, we demonstrated different types of insertion procedures for lipid-anchored protein into membranes. In the liposome sample, we were able to observe the cross-polarization and the (13)C-(13)C chemical shift correlation spectra under MAS, indicating that the liposome sample can be used to analyze molecular interactions using dipolar-based NMR experiments. In contrast, the bicelle sample showed sufficient quality of spectra through scalar-based experiments. The relaxation times and protein-membrane interaction were capable of being analyzed in the bicelle sample. These results demonstrated the applicability of two types of sample system to elucidate the roles of lipid-anchors in regulating diverse biological phenomena.

  2. Incorporation of Ceramides into Saccharomyces cerevisiae Glycosylphosphatidylinositol-Anchored Proteins Can Be Monitored In Vitro▿

    Science.gov (United States)

    Bosson, Régine; Guillas, Isabelle; Vionnet, Christine; Roubaty, Carole; Conzelmann, Andreas

    2009-01-01

    After glycosylphosphatidylinositols (GPIs) are added to GPI proteins of Saccharomyces cerevisiae, a fatty acid of the diacylglycerol moiety is exchanged for a C26:0 fatty acid through the subsequent actions of Per1 and Gup1. In most GPI anchors this modified diacylglycerol-based anchor is subsequently transformed into a ceramide-containing anchor, a reaction which requires Cwh43. Here we show that the last step of this GPI anchor lipid remodeling can be monitored in microsomes. The assay uses microsomes from cells that have been grown in the presence of myriocin, a compound that blocks the biosynthesis of dihydrosphingosine (DHS) and thus inhibits the biosynthesis of ceramide-based anchors. Such microsomes, when incubated with [3H]DHS, generate radiolabeled, ceramide-containing anchor lipids of the same structure as made by intact cells. Microsomes from cwh43Δ or mcd4Δ mutants, which are unable to make ceramide-based anchors in vivo, do not incorporate [3H]DHS into anchors in vitro. Moreover, gup1Δ microsomes incorporate [3H]DHS into the same abnormal anchor lipids as gup1Δ cells synthesize in vivo. Thus, the in vitro assay of ceramide incorporation into GPI anchors faithfully reproduces the events that occur in mutant cells. Incorporation of [3H]DHS into GPI proteins is observed with microsomes alone, but the reaction is stimulated by cytosol or bovine serum albumin, ATP plus coenzyme A (CoA), or C26:0-CoA, particularly if microsomes are depleted of acyl-CoA. Thus, [3H]DHS cannot be incorporated into proteins in the absence of acyl-CoA. PMID:19074599

  3. Incorporation of ceramides into Saccharomyces cerevisiae glycosylphosphatidylinositol-anchored proteins can be monitored in vitro.

    Science.gov (United States)

    Bosson, Régine; Guillas, Isabelle; Vionnet, Christine; Roubaty, Carole; Conzelmann, Andreas

    2009-03-01

    After glycosylphosphatidylinositols (GPIs) are added to GPI proteins of Saccharomyces cerevisiae, a fatty acid of the diacylglycerol moiety is exchanged for a C(26:0) fatty acid through the subsequent actions of Per1 and Gup1. In most GPI anchors this modified diacylglycerol-based anchor is subsequently transformed into a ceramide-containing anchor, a reaction which requires Cwh43. Here we show that the last step of this GPI anchor lipid remodeling can be monitored in microsomes. The assay uses microsomes from cells that have been grown in the presence of myriocin, a compound that blocks the biosynthesis of dihydrosphingosine (DHS) and thus inhibits the biosynthesis of ceramide-based anchors. Such microsomes, when incubated with [(3)H]DHS, generate radiolabeled, ceramide-containing anchor lipids of the same structure as made by intact cells. Microsomes from cwh43Delta or mcd4Delta mutants, which are unable to make ceramide-based anchors in vivo, do not incorporate [(3)H]DHS into anchors in vitro. Moreover, gup1Delta microsomes incorporate [(3)H]DHS into the same abnormal anchor lipids as gup1Delta cells synthesize in vivo. Thus, the in vitro assay of ceramide incorporation into GPI anchors faithfully reproduces the events that occur in mutant cells. Incorporation of [(3)H]DHS into GPI proteins is observed with microsomes alone, but the reaction is stimulated by cytosol or bovine serum albumin, ATP plus coenzyme A (CoA), or C(26:0)-CoA, particularly if microsomes are depleted of acyl-CoA. Thus, [(3)H]DHS cannot be incorporated into proteins in the absence of acyl-CoA.

  4. Endocytosis of GPI-anchored proteins in human lymphocytes: role of glycolipid-based domains, actin cytoskeleton, and protein kinases

    OpenAIRE

    1996-01-01

    GPI-anchored surface proteins mediate many important functions, including transport, signal transduction, adhesion, and protection against complement. They cluster into glycolipid-based membrane domains and caveolae, plasmalemmal vesicles involved in the transcytosis and endocytosis of these surface proteins. However, in lymphocytes, neither the characteristic flask shaped caveolae nor caveolin, a transmembrane protein typical of caveolae, have been observed. Here, we show that the GPI-anchor...

  5. Anchoring Intrinsically Disordered Proteins to Multiple Targets: Lessons from N-Terminus of the p53 Protein

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    Yongqi Huang

    2011-02-01

    Full Text Available Anchor residues, which are deeply buried upon binding, play an important role in protein–protein interactions by providing recognition specificity and facilitating the binding kinetics. Up to now, studies on anchor residues have been focused mainly on ordered proteins. In this study, we investigated anchor residues in intrinsically disordered proteins (IDPs which are flexible in the free state. We identified the anchor residues of the N-terminus of the p53 protein (Glu17–Asn29, abbreviated as p53N which are involved in binding with two different targets (MDM2 and Taz2, and analyzed their side chain conformations in the unbound states. The anchor residues in the unbound p53N were found to frequently sample conformations similar to those observed in the bound complexes (i.e., Phe19, Trp23, and Leu26 in the p53N-MDM2 complex, and Leu22 in the p53N-Taz2 complex. We argue that the bound-like conformations of the anchor residues in the unbound state are important for controlling the specific interactions between IDPs and their targets. Further, we propose a mechanism to account for the binding promiscuity of IDPs in terms of anchor residues and molecular recognition features (MoRFs.

  6. Proteins containing an uncleaved signal for glycophosphatidylinositol membrane anchor attachment are retained in a post-ER compartment

    OpenAIRE

    1992-01-01

    Glycophosphatidylinositol (GPI)-anchored membrane proteins are initially synthesized with a cleavable COOH-terminal extension that signals anchor attachment. Overexpression in COS cells of hGH-DAF fusion proteins containing the GPI signal of decay accelerating factor (DAF) fused to the COOH-terminus of human growth hormone (hGH), produces both GPI-anchored hGH-DAF and uncleaved precursors that retain the GPI signal. Using hGH-DAF fusion proteins containing a mutated, noncleavable GPI signal, ...

  7. Current perspective on protein S-acylation in plants: more than just a fatty anchor?

    Science.gov (United States)

    Hurst, Charlotte H; Hemsley, Piers A

    2015-03-01

    Membranes are an important signalling platform in plants. The plasma membrane is the point where information about the external environment must be converted into intracellular signals, while endomembranes are important sites of protein trafficking, organization, compartmentalization, and intracellular signalling. This requires co-ordinating the spatial distribution of proteins, their activation state, and their interacting partners. This regulation frequently occurs through post-translational modification of proteins. Proteins that associate with the cell membrane do so through transmembrane domains, protein-protein interactions, lipid binding motifs/domains or use the post-translational addition of lipid groups as prosthetic membrane anchors. S-acylation is one such lipid modification capable of anchoring proteins to the membrane. Our current knowledge of S-acylation function in plants is fairly limited compared with other post-translational modifications and S-acylation in other organisms. However, it is becoming increasingly clear that S-acylation can act as more than just a simple membrane anchor: it can also act as a regulatory mechanism in signalling pathways in plants. S-acylation is, therefore, an ideal mechanism for regulating protein function at membranes. This review discusses our current knowledge of S-acylated proteins in plants, the interaction of different lipid modifications, and the general effects of S-acylation on cellular function.

  8. Hypomorphic mutations in PGAP2, encoding a GPI-anchor-remodeling protein, cause autosomal-recessive intellectual disability

    DEFF Research Database (Denmark)

    Hansen, Lars; Tawamie, Hasan; Murakami, Yoshiko

    2013-01-01

    PGAP2 encodes a protein involved in remodeling the glycosylphosphatidylinositol (GPI) anchor in the Golgi apparatus. After synthesis in the endoplasmic reticulum (ER), GPI anchors are transferred to the proteins and are remodeled while transported through the Golgi to the cell membrane. Germline...... rescue when we used strong promoters before the mutant cDNAs, suggesting a hypomorphic effect of the mutations. We report on alterations in the Golgi-located part of the GPI-anchor-biosynthesis pathway and extend the phenotypic spectrum of the GPI-anchor deficiencies to isolated intellectual disability...

  9. Palmitylation of an amino-terminal cysteine motif of protein tyrosine kinases p56lck and p59fyn mediates interaction with glycosyl-phosphatidylinositol-anchored proteins.

    OpenAIRE

    1993-01-01

    Cross-linking of glycosyl-phosphatidylinositol (GPI)-anchored membrane proteins on T cells can trigger cell activation. We and others have shown an association between GPI-anchored proteins and the protein tyrosine kinases (PTKs) p56lck and p59fyn, suggesting a pathway for signaling through GPI-anchored proteins. Studies of decay-accelerating factor (DAF) or CD59 in either the C32 cell line or the HeLa cell line transfected with PTK cDNA demonstrated that the GPI-anchored proteins associated ...

  10. How curved membranes recruit amphipathic helices and protein anchoring motifs

    DEFF Research Database (Denmark)

    Hatzakis, Nikos; Bhatia, Vikram Kjøller; Larsen, Jannik;

    2009-01-01

    Lipids and several specialized proteins are thought to be able to sense the curvature of membranes (MC). Here we used quantitative fluorescence microscopy to measure curvature-selective binding of amphipathic motifs on single liposomes 50-700 nm in diameter. Our results revealed that sensing...

  11. Lactobacillus acidophilus CP23 with weak immunomodulatory activity lacks anchoring structure for surface layer protein.

    Science.gov (United States)

    Yanagihara, Sae; Kato, Shinji; Ashida, Nobuhisa; Yamamoto, Naoyuki

    2015-05-01

    To determine the reason for the low levels of Surface layer protein A (SlpA) on CP23 cells, which might play a crucial role in the immunomodulatory effect of Lactobacillus acidophilus, the DNA sequence of the slpA gene of CP23 and L-92 strains, including the upstream region, were analyzed. Unexpectedly, there was no significant difference in the predicted amino acid sequence of the C-terminus needed for cell anchoring, and only an additional Ala-Val-Ala sequence inserted in the N-terminal region of the mature CP23 protein. Therefore, anchoring of SlpA on the cell wall of CP23 and L-92 was evaluated by a reconstitution assay, which showed that SlpA released by LiCl treatment from both CP23 and L-92 was successfully anchored on LiCl-treated L-92 cells, but not on LiCl-treated CP23 cells. Moreover, quantitative analysis of SlpA protein in the culture medium of CP23 and L-92 by ELISA revealed higher levels of SlpA secretion in CP23 cells than in L-92 cells. Collectively, these results suggest that the lower levels of SlpA on the surface of CP23 cells might be caused by less cell wall capacity for SlpA anchoring, leading to an accumulation of SlpA in the culture medium of CP23 cells. The present study supports the importance of cell surface structure of L. acidophilus L-92 for SlpA anchoring on the cell surface needed for immunomodulatory effect.

  12. Is hexamerin receptor a GPI-anchored protein in Achaea janata (Lepidoptera: Noctuidae)?

    Indian Academy of Sciences (India)

    Madhusudhan Budatha; Thuirei Jacob Ningshen; Aparna Dutta-Gupta

    2011-08-01

    The process of uptake of hexamerins during metamorphosis from insect haemolymph by fat body cells is reminiscent of receptor-mediated endocytosis. Previously, we had identified a hexamerin-binding protein (HBP) and reported for the first time that uptake of hexamerins is dependent on the phosphorylation of HBP partly by a tyrosine kinase, which is, in turn, activated by 20-hydroxyecdysone (20E). However, the exact nature of HBP and the mechanism of interaction are still unknown. Here we report the possibility of HBP being a GPI-anchored protein in the fat body of Achaea janata and its role in the tyrosine-kinase-mediated phosphorylation signalling. Digestion of fat body membrane preparation with bacterial phosphatidylinositol-specific phospholipase C (PI-PLC), and the subsequent recognition by antibodies specific for the cross-reacting determinant (CRD), revealed that HBP is glycosylphosphatidylinositol (GPI)-anchored protein and, further, that the hexamerin binding to HBP was inhibited after digestion. Hexamerin overlay assay (HOA) of co-immunoprecipitated in vitro phosphorylated HBP showed exclusive binding to ∼120 kDa protein. Lectin-binding analysis of hexamerins revealed the presence of -acetylgalactosamine (GalNAc) and -acetylglucosamine (GluNAc), whereas HBP showed the presence of GalNac alone. Mild chemical deglycosylation studies and binding interaction in the presence of sugars revealed that glycan moieties are possibly not involved in the interaction between HBP and hexamerins. Taken together, these results suggest that HBP may be a GPI-anchored protein, and interaction and activation of HBP is through lipid-linked non-receptor src tyrosine kinases. However, additional studies are needed to prove that HBP is a GPI-anchored protein.

  13. A Proteomics Investigation of Anchored PKA-RI Signaling

    NARCIS (Netherlands)

    Kovanich, D.

    2013-01-01

    Compartmentalization of kinases and phosphatases plays an important role in the specificity of second messenger mediated signaling events. Localization of the cAMP-dependent protein kinase is mediated by interaction of its regulatory subunit (PKA-R) with the versatile family of A-kinase anchoring pr

  14. Determination of the non-ionic detergent insolubility and phosphoprotein associations of glycosylphosphatidylinositol-anchored proteins expressed on T cells.

    Science.gov (United States)

    Solomon, K R; Mallory, M A; Finberg, R W

    1998-09-01

    Glycosylphosphatidylinositol (GPI)-anchored proteins are poorly solublized in non-ionic detergents such as Triton X-100 and Nonidet P40, but are easily solublized by detergents with high critical micelle concentrations such as octylglucoside. This solubility profile has been suggested to be due to the localization of GPI-anchored proteins to lipid microdomains rich in cholesterol and sphingolipids. Additionally, GPI-anchored proteins expressed on haemopoietic cells have been shown to associate with src-family tyrosine kinases and heterotrimeric G proteins. Despite these observations, the non-ionic detergent insolubility of GPI-anchored proteins on haemopoietic cells has not been quantified nor has a relationship between the non-ionic detergent insolubility of these proteins and their association with signal-transduction molecules been identified. Here we show that GPI-anchored proteins found on T-cell tumours and activated T cells, although significantly more insoluble then transmembrane proteins, are not uniform in their detergent insolubility. Whereas CD59 was between 4% and 13% soluble, CD48 was between 13% and 25% soluble, CD55 was between 20% and 30% soluble, and CD109 was between 34% and 75% soluble. The ability of these GPI-anchored proteins to associate with phosphoproteins was correlated with their detergent insolubility: the more detergent-insoluble that a GPI-anchored protein was, the greater the level of phosphoprotein associations. These experiments reveal a relationship between non-ionic detergent insolubility and association with signal-transduction molecules and suggest a cause-and-effect relationship between these two properties. In total, these experiments support the hypothesis that the association of GPI-anchored proteins with signalling molecules is due to their sorting to lipid microdomains.

  15. High-throughput sorting of the highest producing cell via a transiently protein-anchored system.

    Directory of Open Access Journals (Sweden)

    Kuo-Hsiang Chuang

    Full Text Available Developing a high-throughput method for the effecient selection of the highest producing cell is very important for the production of recombinant protein drugs. Here, we developed a novel transiently protein-anchored system coupled with fluorescence activated cell sorting (FACS for the efficient selection of the highest producing cell. A furin cleavage peptide (RAKR was used to join a human anti-epithelial growth factor antibody (αEGFR Ab and the extracellular-transmembrane-cytosolic domains of the mouse B7-1 antigen (B7. The furin inhibitor can transiently switch secreted αEGFR Ab into a membrane-anchored form. After cell sorting, the level of membrane αEGFR Ab-RAKR-B7 is proportional to the amount of secreted αEGFR Ab in the medium. We further selected 23 αEGFR Ab expressing cells and demonstrated a high correlation (R2 = 0.9165 between the secretion level and surface expression levels of αEGFR Ab. These results suggested that the novel transiently protein-anchored system can easily and efficiently select the highest producing cells, reducing the cost for the production of biopharmaceuticals.

  16. Defects in RGS9 or its anchor protein R9AP in patients with slow photoreceptor deactivation

    NARCIS (Netherlands)

    Nishiguchi, KM; Sandberg, MA; Kooijman, AC; Martemyanov, KA; Pott, JWR; Hagstrom, SA; Arshavsky, VY; Berson, EL; Dryja, TP

    2004-01-01

    The RGS proteins are GTPase activating proteins that accelerate the deactivation of G proteins in a variety of signalling pathways in eukaryotes(1-6). RGS9 deactivates the G proteins (transducins) in the rod and cone phototransduction cascades(7,8). It is anchored to photoreceptor membranes by the t

  17. GPI-anchored proteins do not reside in ordered domains in the live cell plasma membrane

    Science.gov (United States)

    Sevcsik, Eva; Brameshuber, Mario; Fölser, Martin; Weghuber, Julian; Honigmann, Alf; Schütz, Gerhard J.

    2015-04-01

    The organization of proteins and lipids in the plasma membrane has been the subject of a long-lasting debate. Membrane rafts of higher lipid chain order were proposed to mediate protein interactions, but have thus far not been directly observed. Here we use protein micropatterning combined with single-molecule tracking to put current models to the test: we rearranged lipid-anchored raft proteins (glycosylphosphatidylinositol(GPI)-anchored-mGFP) directly in the live cell plasma membrane and measured the effect on the local membrane environment. Intriguingly, this treatment does neither nucleate the formation of an ordered membrane phase nor result in any enrichment of nanoscopic-ordered domains within the micropatterned regions. In contrast, we find that immobilized mGFP-GPIs behave as inert obstacles to the diffusion of other membrane constituents without influencing their membrane environment over distances beyond their physical size. Our results indicate that phase partitioning is not a fundamental element of protein organization in the plasma membrane.

  18. Loss of Dfg5 glycosylphosphatidylinositol-anchored membrane protein confers enhanced heat tolerance in Saccharomyces cerevisiae.

    Science.gov (United States)

    Nasution, Olviyani; Lee, Jaok; Srinivasa, Kavitha; Choi, In-Geol; Lee, Young Mi; Kim, Eunjung; Choi, Wonja; Kim, Wankee

    2015-08-01

    The protein product of Saccharomyces cerevisiae DFG5 gene is a glycosylphosphatidylinositol (GPI)-anchored plasma membrane protein and a putative glycosidase/glycosyltransferase that links other GPI-anchored proteins to β-glucans in the cell wall. Upon exposure to heat (41°C), DFG5 deletion mutant dfg5Δ displayed significantly enhanced heat tolerance as well as lowered level of reactive oxygen species and decreased membrane permeability compared with those in the control (BY4741). Comparative transcriptome profiles of BY4741 and dfg5Δ revealed that 38 and 23 genes were up- and down-regulated in dfg5Δ respectively. Of the 23 down-regulated genes, 11 of 13 viable deletion mutants were identified to be tolerant to heat, suggesting that the down-regulation of those genes might have contributed to the enhanced heat tolerance in dfg5Δ. Deletion of DFG5 caused slight activation of mitogen-activated protein kinases Hog1 in the high-osmolarity glycerol pathway and Slt2 in the cell wall integrity pathway. Therefore, a model is proposed on the signal transduction pathways associated with deletion of DFG5 upon heat stress.

  19. Signal transduction across cellular membranes can be mediated by coupling of the clustering of anchored proteins in both leaflets

    Science.gov (United States)

    Yue, Tongtao; Zhang, Xianren

    2012-01-01

    One key question in signal transduction is how the signal is relayed from the outer leaflet of a cellular membrane to the inner leaflet. Using a simulation model, a mechanism for the mediation of signal transduction is proposed here in which the coupling between membrane proteins in different leaflets can be achieved by the clustering of anchored proteins, without recruiting transmembrane proteins. Depending on the hydrophobic length of the anchored proteins, three coupling patterns, including face-to-face clustering, interdigitated clustering, and weak-coupled clustering, are observed in this work. This observation provides a possible explanation of how a particular downstream signaling pathway is selected.

  20. The association of BAG6 with SGTA and tail-anchored proteins.

    Directory of Open Access Journals (Sweden)

    Pawel Leznicki

    Full Text Available BACKGROUND: The BAG6 protein is a subunit of a heterotrimeric complex that binds a range of membrane and secretory protein precursors localized to the cytosol, enforcing quality control and influencing their subsequent fate. METHODOLOGY AND PRINCIPAL FINDINGS: BAG6 has an N-terminal ubiquitin-like domain, and a C-terminal Bcl-2-associated athanogene domain, separated by a large central proline-rich region. We have used in vitro binding approaches to identify regions of BAG6 important for its interactions with: i the small-glutamine rich tetratricopeptide repeat-containing protein alpha (SGTA and ii two model tail-anchored membrane proteins as a paradigm for its hydrophobic substrates. We show that the BAG6-UBL is essential for binding to SGTA, and find that the UBL of a second subunit of the BAG6-complex, ubiquitin-like protein 4A (UBL4A, competes for SGTA binding. Our data show that this binding is selective, and suggest that SGTA can bind either BAG6, or UBL4A, but not both at the same time. We adapted our in vitro binding assay to study the association of BAG6 with an immobilized tail-anchored protein, Sec61β, and find both the UBL and BAG domains are dispensable for binding this substrate. This conclusion was further supported using a heterologous subcellular localization assay in yeast, where the BAG6-dependent nuclear relocalization of a second tail-anchored protein, GFP-Sed5, also required neither the UBL, nor the BAG domain of BAG6. SIGNIFICANCE: On the basis of these findings, we propose a working model where the large central region of the BAG6 protein provides a binding site for a diverse group of substrates, many of which expose a hydrophobic stretch of polypeptide. This arrangement would enable the BAG6 complex to bring together its substrates with potential effectors including those recruited via its N-terminal UBL. Such effectors may include SGTA, and the resulting assemblies influence the subsequent fate of the hydrophobic BAG6

  1. Generation of glycosylphosphatidylinositol anchor protein-deficient blood cells from human induced pluripotent stem cells.

    Science.gov (United States)

    Yuan, Xuan; Braunstein, Evan M; Ye, Zhaohui; Liu, Cyndi F; Chen, Guibin; Zou, Jizhong; Cheng, Linzhao; Brodsky, Robert A

    2013-11-01

    PIG-A is an X-linked gene required for the biosynthesis of glycosylphosphatidylinositol (GPI) anchors; thus, PIG-A mutant cells have a deficiency or absence of all GPI-anchored proteins (GPI-APs). Acquired mutations in hematopoietic stem cells result in the disease paroxysmal nocturnal hemoglobinuria, and hypomorphic germline PIG-A mutations lead to severe developmental abnormalities, seizures, and early death. Human induced pluripotent stem cells (iPSCs) can differentiate into cell types derived from all three germ layers, providing a novel developmental system for modeling human diseases. Using PIG-A gene targeting and an inducible PIG-A expression system, we have established, for the first time, a conditional PIG-A knockout model in human iPSCs that allows for the production of GPI-AP-deficient blood cells. PIG-A-null iPSCs were unable to generate hematopoietic cells or any cells expressing the CD34 marker and were defective in generating mesodermal cells expressing KDR/VEGFR2 (kinase insert domain receptor) and CD56 markers. In addition, PIG-A-null iPSCs had a block in embryonic development prior to mesoderm differentiation that appears to be due to defective signaling through bone morphogenetic protein 4. However, early inducible PIG-A transgene expression allowed for the generation of GPI-AP-deficient blood cells. This conditional PIG-A knockout model should be a valuable tool for studying the importance of GPI-APs in hematopoiesis and human development.

  2. Molecular view on protein sorting into liquid-ordered membrane domains mediated by gangliosides and lipid anchors

    NARCIS (Netherlands)

    de Jong, Djurre H.; Lopez, Cesar A.; Marrink, Siewert J.

    2013-01-01

    We present results from coarse grain molecular dynamics simulations of mixed model membranes consisting of saturated and unsaturated lipids together with cholesterol, in which lipid-anchored membrane proteins are embedded. The membrane proteins studied are the peripherally bound H-Ras, N-Ras, and He

  3. The role of Listeria monocytogenes cell wall surface anchor protein LapB in virulence, adherence, and intracellular replication

    Science.gov (United States)

    Lmof2365_2117 is a Listeria monocytogenes putative cell wall surface anchor protein with a conserved domain found in collagen binding proteins. We constructed a deletion mutation in lmof2365_2117 in serotype 4b strain F2365, evaluated its virulence, and determined its ability to adhere and invade co...

  4. Mammalian carboxylesterase (CES) releases GPI-anchored proteins from the cell surface upon lipid raft fluidization.

    Science.gov (United States)

    Orihashi, Kaoru; Tojo, Hiromasa; Okawa, Katsuya; Tashima, Yuko; Morita, Takashi; Kondoh, Gen

    2012-03-01

    Mammalian carboxylesterase (CES) is well known as a biotransformation enzyme for prodrugs and xenobiotics. Here, we purified CES as a GPI-anchored protein (GPI-AP)-releasing factor (GPIase) that releases such protein from the cell surface. All five isoforms of CES showed this activity to various degrees. When the serine residue of the catalytic triad for esterase was replaced by alanine, esterase activity was completely disrupted, while full GPIase activity remained, suggesting that these two activities are exhibited via different mechanisms. CES6, a new class of mammalian CES, exhibited the highest GPIase activity and released specific GPI-APs from the cell surface after lipid raft fluidization. The released product contained a GPI component, indicating that GPI-AP was released by cleavage in GPI. These results revealed for the first time that CES recognizes and catalyzes macromolecule GPI-AP as well as small molecules.

  5. A novel family of Apicomplexan glideosome-associated proteins with an inner membrane-anchoring role.

    Science.gov (United States)

    Bullen, Hayley E; Tonkin, Christopher J; O'Donnell, Rebecca A; Tham, Wai-Hong; Papenfuss, Anthony T; Gould, Sven; Cowman, Alan F; Crabb, Brendan S; Gilson, Paul R

    2009-09-11

    The phylum Apicomplexa are a group of obligate intracellular parasites responsible for a wide range of important diseases. Central to the lifecycle of these unicellular parasites is their ability to migrate through animal tissue and invade target host cells. Apicomplexan movement is generated by a unique system of gliding motility in which substrate adhesins and invasion-related proteins are pulled across the plasma membrane by an underlying actin-myosin motor. The myosins of this motor are inserted into a dual membrane layer called the inner membrane complex (IMC) that is sandwiched between the plasma membrane and an underlying cytoskeletal basket. Central to our understanding of gliding motility is the characterization of proteins residing within the IMC, but to date only a few proteins are known. We report here a novel family of six-pass transmembrane proteins, termed the GAPM family, which are highly conserved and specific to Apicomplexa. In Plasmodium falciparum and Toxoplasma gondii the GAPMs localize to the IMC where they form highly SDS-resistant oligomeric complexes. The GAPMs co-purify with the cytoskeletal alveolin proteins and also to some degree with the actin-myosin motor itself. Hence, these proteins are strong candidates for an IMC-anchoring role, either directly or indirectly tethering the motor to the cytoskeleton.

  6. Vaccination with Enzymatically Cleaved GPI-Anchored Proteins from Schistosoma mansoni Induces Protection against Challenge Infection

    Directory of Open Access Journals (Sweden)

    Vicente P. Martins

    2012-01-01

    Full Text Available The flatworm Schistosoma mansoni is a blood fluke parasite that causes schistosomiasis, a debilitating disease that occurs throughout the developing world. Current schistosomiasis control strategies are mainly based on chemotherapy, but many researchers believe that the best long-term strategy to control schistosomiasis is through immunization with an antischistosomiasis vaccine combined with drug treatment. In the search for potential vaccine candidates, numerous tegument antigens have been assessed. As the major interface between parasite and mammalian host, the tegument plays crucial roles in the establishment and further course of schistosomiasis. Herein, we evaluated the potential of a GPI fraction, containing representative molecules located on the outer surface of adult worms, as vaccine candidate. Immunization of mice with GPI-anchored proteins induced a mixed Th1/Th2 type of immune response with production of IFN-γ and TNF-α, and low levels of IL-5 into the supernatant of splenocyte cultures. The protection engendered by this vaccination protocol was confirmed by 42% reduction in worm burden, 45% reduction in eggs per gram of hepatic tissue, 29% reduction in the number of granulomas per area, and 53% reduction in the granuloma fibrosis. Taken together, the data herein support the potential of surface-exposed GPI-anchored antigens from the S. mansoni tegument as vaccine candidate.

  7. Characterisation of a cell wall-anchored protein of Staphylococcus saprophyticus associated with linoleic acid resistance

    Directory of Open Access Journals (Sweden)

    King Nathan P

    2012-01-01

    Full Text Available Abstract Background The Gram-positive bacterium Staphylococcus saprophyticus is the second most frequent causative agent of community-acquired urinary tract infections (UTI, accounting for up to 20% of cases. A common feature of staphylococci is colonisation of the human skin. This involves survival against innate immune defenses including antibacterial unsaturated free fatty acids such as linoleic acid which act by disrupting bacterial cell membranes. Indeed, S. saprophyticus UTI is usually preceded by perineal skin colonisation. Results In this study we identified a previously undescribed 73.5 kDa cell wall-anchored protein of S. saprophyticus, encoded on plasmid pSSAP2 of strain MS1146, which we termed S. saprophyticus surface protein F (SssF. The sssF gene is highly prevalent in S. saprophyticus clinical isolates and we demonstrate that the SssF protein is expressed at the cell surface. However, unlike all other characterised cell wall-anchored proteins of S. saprophyticus, we were unable to demonstrate a role for SssF in adhesion. SssF shares moderate sequence identity to a surface protein of Staphylococcus aureus (SasF recently shown to be an important mediator of linoleic acid resistance. Using a heterologous complementation approach in a S. aureus sasF null genetic background, we demonstrate that SssF is associated with resistance to linoleic acid. We also show that S. saprophyticus strains lacking sssF are more sensitive to linoleic acid than those that possess it. Every staphylococcal genome sequenced to date encodes SssF and SasF homologues. Proteins in this family share similar predicted secondary structures consisting almost exclusively of α-helices in a probable coiled-coil formation. Conclusions Our data indicate that SssF is a newly described and highly prevalent surface-localised protein of S. saprophyticus that contributes to resistance against the antibacterial effects of linoleic acid. SssF is a member of a protein family

  8. A unifying mechanism accounts for sensing of membrane curvature by BAR domains, amphipathic helices and membrane-anchored proteins

    DEFF Research Database (Denmark)

    Bhatia, Vikram Kjøller; Hatzakis, Nikos; Stamou, Dimitrios

    2010-01-01

    unrelated motifs: BAR domains, amphipathic helices and membrane-anchored proteins. We discuss the conclusion that the curvature of the BAR dimer is not responsible for sensing and that the sensing properties of all three motifs can be rationalized by the physicochemical properties of the curved membrane......The discovery of proteins that recognize membrane curvature created a paradigm shift by suggesting that membrane shape may act as a cue for protein localization that is independent of lipid or protein composition. Here we review recent data on membrane curvature sensing by three structurally...... itself. We thus anticipate that membrane curvature will promote the redistribution of proteins that are anchored in membranes through any type of hydrophobic moiety, a thesis that broadens tremendously the implications of membrane curvature for protein sorting, trafficking and signaling in cell biology....

  9. Multiple functions of inositolphosphorylceramides in the formation and intracellular transport of glycosylphosphatidylinositol-anchored proteins in yeast.

    Science.gov (United States)

    Bosson, Régine; Conzelmann, Andreas

    2007-01-01

    The mature sphingolipids of yeast consist of IPCs (inositolphosphorylceramides) and glycosylated derivatives thereof. Beyond being an abundant membrane constituent in the organelles of the secretory pathway, IPCs are also used to constitute the lipid moiety of the majority of GPI (glycosylphosphatidylinositol) proteins, while a minority of GPI proteins contain PI (phosphatidylinositol). Thus all GPI anchor lipids (as well as free IPCs) typically contain C26 fatty acids. However, the primary GPI lipid that isadded to newly synthesized proteins in the endoplasmic reticulum consists of a PI with conventional C16 and C18 fatty acids. A new class of enzymes is required to replace the fatty acid in sn-2 by a C26 fatty acid. Cells lacking this activity make normal amounts of GPI proteins but accumulate GPI anchors containing lyso-PI. As a consequence, the endoplasmic reticulum to Golgi transport of the GPI protein Gas1p is slow, and mature Gas1p is lost from the plasma membrane into the medium. The GPI anchor containing C26 in sn-2 can further be remodelled by the exchange of diacylglycerol for ceramide. This process is also dependent on the presence of specific phosphorylethanolamine side-chains on the GPI anchor.

  10. The surface-anchored NanA protein promotes pneumococcal brain endothelial cell invasion.

    Science.gov (United States)

    Uchiyama, Satoshi; Carlin, Aaron F; Khosravi, Arya; Weiman, Shannon; Banerjee, Anirban; Quach, Darin; Hightower, George; Mitchell, Tim J; Doran, Kelly S; Nizet, Victor

    2009-08-31

    In humans, Streptococcus pneumoniae (SPN) is the leading cause of bacterial meningitis, a disease with high attributable mortality and frequent permanent neurological sequelae. The molecular mechanisms underlying the central nervous system tropism of SPN are incompletely understood, but include a primary interaction of the pathogen with the blood-brain barrier (BBB) endothelium. All SPN strains possess a gene encoding the surface-anchored sialidase (neuraminidase) NanA, which cleaves sialic acid on host cells and proteins. Here, we use an isogenic SPN NanA-deficient mutant and heterologous expression of the protein to show that NanA is both necessary and sufficient to promote SPN adherence to and invasion of human brain microvascular endothelial cells (hBMECs). NanA-mediated hBMEC invasion depends only partially on sialidase activity, whereas the N-terminal lectinlike domain of the protein plays a critical role. NanA promotes SPN-BBB interaction in a murine infection model, identifying the protein as proximal mediator of CNS entry by the pathogen.

  11. Distinct pathways mediate the sorting of tail-anchored proteins to the plastid outer envelope.

    Directory of Open Access Journals (Sweden)

    Preetinder K Dhanoa

    Full Text Available BACKGROUND: Tail-anchored (TA proteins are a distinct class of membrane proteins that are sorted post-translationally to various organelles and function in a number of important cellular processes, including redox reactions, vesicular trafficking and protein translocation. While the molecular targeting signals and pathways responsible for sorting TA proteins to their correct intracellular destinations in yeasts and mammals have begun to be characterized, relatively little is known about TA protein biogenesis in plant cells, especially for those sorted to the plastid outer envelope. METHODOLOGY/PRINCIPAL FINDINGS: Here we investigated the biogenesis of three plastid TA proteins, including the 33-kDa and 34-kDa GTPases of the translocon at the outer envelope of chloroplasts (Toc33 and Toc34 and a novel 9-kDa protein of unknown function that we define here as an outer envelope TA protein (OEP9. Using a combination of in vivo and in vitro assays we show that OEP9 utilizes a different sorting pathway than that used by Toc33 and Toc34. For instance, while all three TA proteins interact with the cytosolic OEP chaperone/receptor, AKR2A, the plastid targeting information within OEP9 is distinct from that within Toc33 and Toc34. Toc33 and Toc34 also appear to differ from OEP9 in that their insertion is dependent on themselves and the unique lipid composition of the plastid outer envelope. By contrast, the insertion of OEP9 into the plastid outer envelope occurs in a proteinaceous-dependent, but Toc33/34-independent manner and membrane lipids appear to serve primarily to facilitate normal thermodynamic integration of this TA protein. CONCLUSIONS/SIGNIFICANCE: Collectively, the results provide evidence in support of at least two sorting pathways for plastid TA outer envelope proteins and shed light on not only the complex diversity of pathways involved in the targeting and insertion of proteins into plastids, but also the molecular mechanisms that underlie

  12. ADAM12 is expressed in the tumour vasculature and mediates ectodomain shedding of several membrane-anchored endothelial proteins

    DEFF Research Database (Denmark)

    Frohlich, Camilla; Klitgaard, Marie; Noer, Julie B

    2013-01-01

    ADAM (a disintegrin and metalloproteinase) 12 is a metalloprotease implicated in cancer progression. ADAM12 can activate membrane-anchored proteins, such as sonic hedgehog, Delta-like 1 and certain epidermal growth factor receptor ligands, through a process called ectodomain shedding. We screened...

  13. QM/MD studies of the dynamics of the MTSL spin label in Aurora-A kinase protein activation loop

    CERN Document Server

    Concilio, Maria Grazia; Bayliss, Richard; Burgess, Selena

    2015-01-01

    Molecular dynamics(MD)simulations using a graphics processing unit (GPU) has been employed in order to determine the conformational space of the methane-thiosulfonate spin label (MTSL) attached to the activation loop of the Aurora-A kinase protein and compared with quantum mechanical (QM) methods rooted on density functional theory (DFT). MD provided a wealth of information about interactions between the MTSL and the residues of the protein and on the different motional contributions to the overall dynamics of the MTSL. Data obtained from MD were seen to be in good agreement with those obtained from QM but the dynamics of the system revealed more interactions than those observed from QM methods. A strong correlation between the tumbling of the protein and the transitions of the X4 and X5 dihedral angles of the MTSL, was observed with a consequent effect also the distribution of the nitroxide(NO)group in the space. Theoretical EPR spectra calculated from opportunely selected MD frames showing interactions betw...

  14. The lactococcal abortive infection protein AbiP is membrane-anchored and binds nucleic acids.

    Science.gov (United States)

    Domingues, Susana; McGovern, Stephen; Plochocka, Danuta; Santos, Mário A; Ehrlich, S Dusko; Polard, Patrice; Chopin, Marie-Christine

    2008-03-30

    AbiP, a lactococcal abortive phage infection system, has previously been shown to arrest phage bIL66M1 DNA replication around 10 min after infection and to inhibit the switch off of phage early transcripts. We report here the functional characterization and implication in the abortive infection phenotype of two domains identified in the AbiP sequence. We show that AbiP is a protein anchored to the membrane by an N-terminal membrane-spanning domain. Our results further suggest that membrane localization may be required for the anti-phage activity of AbiP. The remainder of the protein, which contains a putative nucleic acid binding domain, is shown to be located on the cytosolic side. Purified AbiP is shown to bind nucleic acids with an approximately 10-fold preference for RNA relative to ssDNA. AbiP interaction with both ssDNA and RNA molecules occurs in a sequence-independent manner. We have analyzed the effect of substitutions of aromatic and basic residues on the surface of the putative binding fold. In vitro and in vivo studies of these AbiP derivatives indicate that the previously reported effects on phage development might be dependent on the nucleic acid binding activity displayed by the membrane-bound protein.

  15. Lipase active-site-directed anchoring of organometallics: metallopincer/protein hybrids.

    Science.gov (United States)

    Kruithof, Cornelis A; Casado, Miguel A; Guillena, Gabriela; Egmond, Maarten R; van der Kerk-van Hoof, Anca; Heck, Albert J R; Klein Gebbink, Robertus J M; van Koten, Gerard

    2005-11-18

    The work described herein presents a strategy for the regioselective introduction of organometallic complexes into the active site of the lipase cutinase. Nitrophenol phosphonate esters, well known for their lipase inhibitory activity, are used as anchor functionalities and were found to be ideal tools to develop a single-site-directed immobilization method. A small series of phosphonate esters, covalently attached to ECE "pincer"-type d8-metal complexes through a propyl tether (ECE=[C6H3(CH2E)(2)-2,6]-; E=NR2 or SR), were designed and synthesized. Cutinase was treated with these organometallic phosphonate esters and the new metal-complex/protein hybrids were identified as containing exactly one organometallic unit per protein. The organometallic proteins were purified by membrane dialysis and analyzed by ESI-mass spectrometry. The major advantages of this strategy are: 1) one transition metal can be introduced regioselectively and, hence, the metal environment can potentially be fine-tuned; 2) purification procedures are facile due to the use of pre-synthesized metal complexes; and, most importantly, 3) the covalent attachment of robust organometallic pincer complexes to an enzyme is achieved, which will prevent metal leaching from these hybrids. The approach presented herein can be regarded as a tool in the development of regio- and enantioselective catalyst as well as analytical probes for studying enzyme properties (e.g., structure) and, hence, is a "proof-of-principle design" study in enzyme chemistry.

  16. Functional convergence of signalling by GPI-anchored and anchorless forms of a salamander protein implicated in limb regeneration

    OpenAIRE

    Blassberg, R. A.; Garza-Garcia, A.; Janmohamed, A.; Gates, P. B.; Brockes, J P

    2011-01-01

    The GPI-anchor is an established determinant of molecular localisation and various functional roles have been attributed to it. The newt GPI-anchored three-finger protein (TFP) Prod1 is an important regulator of cell behaviour during limb regeneration, but it is unclear how it signals to the interior of the cell. Prod1 was expressed by transfection in cultured newt limb cells and activated transcription and expression of matrix metalloproteinase 9 (MMP9) by a pathway involving ligand-independ...

  17. Anchoring of c-myc on nuclear matrix proteins in process of mouse thymic T lymphocyte proliferation induced by ConA

    Institute of Scientific and Technical Information of China (English)

    曾丛梅; 蔡树涛; 周凤兰; 张锦珠; 王平

    1996-01-01

    Isolation and characteriation of functional nudear matrix proteins involved in DNA anchoring and gene expression is one of the major subjects of current nudear matrix research. Southwestern blotting (DNA-protein hybridization) was applied to studying the anchoring of c-myc on the nudear matrix proteins in mouse thymic T lymphocytes. The results showed that c-myc bound to the lamin, p34 and p36 nudear matrix proteins specifically. In the process of mouse thymic PNA T lymphocytes proliferation induced by ConA, the anchoring of c-myc on p34 and p36 nudear matrix proteins changed dynamically.

  18. Clumping factor A, von Willebrand factor-binding protein and von Willebrand factor anchor Staphylococcus aureus to the vessel wall.

    Science.gov (United States)

    Claes, J; Liesenborghs, L; Peetermans, M; Veloso, T R; Missiakas, D; Schneewind, O; Mancini, S; Entenza, J M; Hoylaerts, M F; Heying, R; Verhamme, P; Vanassche, T

    2017-02-09

    Essentials Staphylococcus aureus (S. aureus) binds to endothelium via von Willebrand factor (VWF). Secreted VWF-binding protein (vWbp) mediates S. aureus adhesion to VWF under shear stress. vWbp interacts with VWF and the Sortase A-dependent surface protein Clumping factor A (ClfA). VWF-vWbp-ClfA anchor S. aureus to vascular endothelium under shear stress.

  19. Functional convergence of signalling by GPI-anchored and anchorless forms of a salamander protein implicated in limb regeneration.

    Science.gov (United States)

    Blassberg, Robert A; Garza-Garcia, Acely; Janmohamed, Azara; Gates, Phillip B; Brockes, Jeremy P

    2011-01-01

    The GPI-anchor is an established determinant of molecular localisation and various functional roles have been attributed to it. The newt GPI-anchored three-finger protein (TFP) Prod1 is an important regulator of cell behaviour during limb regeneration, but it is unclear how it signals to the interior of the cell. Prod1 was expressed by transfection in cultured newt limb cells and activated transcription and expression of matrix metalloproteinase 9 (MMP9) by a pathway involving ligand-independent activation of epidermal growth factor receptor (EGFR) signalling and phosphorylation of extracellular regulated kinase 1 and 2 (ERK1/2). This was dependent on the presence of the GPI-anchor and critical residues in the α-helical region of the protein. Interestingly, Prod1 in the axolotl, a salamander species that also regenerates its limbs, was shown to activate ERK1/2 signalling and MMP9 transcription despite being anchorless, and both newt and axolotl Prod1 co-immunoprecipitated with the newt EGFR after transfection. The substitution of the axolotl helical region activated a secreted, anchorless version of the newt molecule. The activity of the newt molecule cannot therefore depend on a unique property conferred by the anchor. Prod1 is a salamander-specific TFP and its interaction with the phylogenetically conserved EGFR has implications for our view of regeneration as an evolutionary variable.

  20. Msp1/ATAD1 maintains mitochondrial function by facilitating the degradation of mislocalized tail-anchored proteins

    Science.gov (United States)

    Chen, Yu-Chan; Umanah, George K E; Dephoure, Noah; Andrabi, Shaida A; Gygi, Steven P; Dawson, Ted M; Dawson, Valina L; Rutter, Jared

    2014-01-01

    The majority of ER-targeted tail-anchored (TA) proteins are inserted into membranes by the Guided Entry of Tail-anchored protein (GET) system. Disruption of this system causes a subset of TA proteins to mislocalize to mitochondria. We show that the AAA+ ATPase Msp1 limits the accumulation of mislocalized TA proteins on mitochondria. Deletion of MSP1 causes the Pex15 and Gos1 TA proteins to accumulate on mitochondria when the GET system is impaired. Likely as a result of failing to extract mislocalized TA proteins, yeast with combined mutation of the MSP1 gene and the GET system exhibit strong synergistic growth defects and severe mitochondrial damage, including loss of mitochondrial DNA and protein and aberrant mitochondrial morphology. Like yeast Msp1, human ATAD1 limits the mitochondrial mislocalization of PEX26 and GOS28, orthologs of Pex15 and Gos1, respectively. GOS28 protein level is also increased in ATAD1−/− mouse tissues. Therefore, we propose that yeast Msp1 and mammalian ATAD1 are conserved members of the mitochondrial protein quality control system that might promote the extraction and degradation of mislocalized TA proteins to maintain mitochondrial integrity. PMID:24843043

  1. Interaction between fidgetin and protein kinase A-anchoring protein AKAP95 is critical for palatogenesis in the mouse.

    Science.gov (United States)

    Yang, Yan; Mahaffey, Connie L; Bérubé, Nathalie; Frankel, Wayne N

    2006-08-01

    The gene defective in fidget mice encodes fidgetin, a member of the AAA (ATPases associated with diverse cellular activities) family of ATPases. Using a yeast two-hybrid screen, we identified cAMP-dependent protein kinase A anchoring protein 95 kDa (AKAP95) as a potential fidgetin-binding protein. Epitope-tagged fidgetin co-localized with endogenous AKAP95 in the nuclear matrix, and the physical interaction between fidgetin and AKAP95 was further confirmed by reciprocal immunoprecipitation. To evaluate the biological significance of the fidgetin-AKAP95 binding, we created AKAP95 mutant mice through a gene trap strategy. Akap95 mutant mice are surprisingly viable with no overt phenotype. However, a significant number of mice carrying both Akap95 and fidget mutations die soon after birth due to cleft palate, consistent with the overlapping expression of AKAP95 and fidgetin in the branchial arches during mouse embryogenesis. These results expand the spectrum of the pleiotropic phenotypes of fidget mice and provide new leads on the in vivo function of AKAP95.

  2. Comparative Haploid Genetic Screens Reveal Divergent Pathways in the Biogenesis and Trafficking of Glycophosphatidylinositol-Anchored Proteins

    Directory of Open Access Journals (Sweden)

    Eric M. Davis

    2015-06-01

    Full Text Available Glycophosphatidylinositol-anchored proteins (GPI-APs play essential roles in physiology, but their biogenesis and trafficking have not been systematically characterized. Here, we took advantage of the recently available haploid genetics approach to dissect GPI-AP pathways in human cells using prion protein (PrP and CD59 as model molecules. Our screens recovered a large number of common and unexpectedly specialized factors in the GPI-AP pathways. PIGN, PGAP2, and PIGF, which encode GPI anchor-modifying enzymes, were selectively isolated in the CD59 screen, suggesting that GPI anchor composition significantly influences the biogenesis of GPI-APs in a substrate-dependent manner. SEC62 and SEC63, which encode components of the ER-targeting machinery, were selectively recovered in the PrP screen, indicating that they do not constitute a universal route for the biogenesis of mammalian GPI-APs. Together, these comparative haploid genetic screens demonstrate that, despite their similarity in overall architecture and subcellular localization, GPI-APs follow markedly distinct biosynthetic and trafficking pathways.

  3. Fatal transmissible amyloid encephalopathy: a new type of prion disease associated with lack of prion protein membrane anchoring.

    Directory of Open Access Journals (Sweden)

    Bruce Chesebro

    2010-03-01

    Full Text Available Prion diseases are fatal neurodegenerative diseases of humans and animals characterized by gray matter spongiosis and accumulation of aggregated, misfolded, protease-resistant prion protein (PrPres. PrPres can be deposited in brain in an amyloid-form and/or non-amyloid form, and is derived from host-encoded protease-sensitive PrP (PrPsen, a protein normally anchored to the plasma membrane by glycosylphosphatidylinositol (GPI. Previously, using heterozygous transgenic mice expressing only anchorless PrP, we found that PrP anchoring to the cell membrane was required for typical clinical scrapie. However, in the present experiments, using homozygous transgenic mice expressing two-fold more anchorless PrP, scrapie infection induced a new fatal disease with unique clinical signs and altered neuropathology, compared to non-transgenic mice expressing only anchored PrP. Brain tissue of transgenic mice had high amounts of infectivity, and histopathology showed dense amyloid PrPres plaque deposits without gray matter spongiosis. In contrast, infected non-transgenic mice had diffuse non-amyloid PrPres deposits with significant gray matter spongiosis. Brain graft studies suggested that anchored PrPsen expression was required for gray matter spongiosis during prion infection. Furthermore, electron and light microscopic studies in infected transgenic mice demonstrated several pathogenic processes not seen in typical prion disease, including cerebral amyloid angiopathy and ultrastructural alterations in perivascular neuropil. These findings were similar to certain human familial prion diseases as well as to non-prion human neurodegenerative diseases, such as Alzheimer's disease.

  4. Phosphorylation of anchoring protein by calmodulin protein kinase associated to the sarcoplasmic reticulum of rabbit fast-twitch muscle.

    Science.gov (United States)

    Damiani, E; Sacchetto, R; Margreth, A

    2000-12-09

    Regulatory phosphorylation of phospholamban and of SR Ca(2+)-ATPase SERCA2a isoform by endogenous CaM-K II in slow-twitch skeletal and cardiac sarcoplasmic reticulum (SR) is well documented, but much less is known of the exact functional role of CaM K II in fast-twitch muscle SR. Recently, it was shown that RNA splicing of brain-specific alpha CaM K II, gives rise to a truncated protein (alpha KAP), consisting mainly of the association domain, serving to anchor CaM K II to SR membrane in rat skeletal muscle [Bayer, K.-U., et al. (1998) EMBO J. 19, 5598-5605]. In the present study, we searched for the presence of alpha KAP in sucrose-density purified SR membrane fractions from representative fast-twitch and slow-twitch limb muscles, both of the rabbit and the rat, using immunoblot techniques and antibody directed against the association domain of alpha CaM K II. Putative alpha KAP was immunodetected as a 23-kDa electrophoretic component on SDS-PAGE of the isolated SR from fast-twitch but not from slow-twitch muscle, and was further identified as a specific substrate of endogenous CaM K II, in the rabbit. Immunodetected, (32)P-labeled, non-calmodulin binding protein, behaved as a single 23-kDa protein species under several electrophoretic conditions. The 23-kDa protein, with defined properties, was isolated as a complex with 60-kDa delta CaM K II isoform, by sucrose-density sedimentation analysis. Moreover, we show here that putative alphaKAP, in spite of its inability to bind CaM in ligand blot overlay, co-eluted with delta CaM K II from CaM-affinity columns. That raises the question of whether CaM K II-mediated phosphorylation of alpha KAP and triadin together might be involved in a molecular signaling pathway important for SR Ca(2+)-release in fast-twitch muscle SR.

  5. Glycosylphosphatidyl Inositol-anchored Proteins and fyn Kinase Assemble in Noncaveolar Plasma Membrane Microdomains Defined by Reggie-1 and -2

    OpenAIRE

    Stürmer, Claudia; Lang, Dirk M.; Kirsch, Friederike; Wiechers, Marianne F.; Deininger, Sören-Oliver; Plattner, Helmut

    2001-01-01

    Using confocal laser scanning and double immunogold electron microscopy, we demonstrate that reggie-1 and -2 are colocalized in ≤0.1-μm plasma membrane microdomains of neurons and astrocytes. In astrocytes, reggie-1 and -2 do not occur in caveolae but clearly outside these structures. Microscopy and coimmunoprecipitation show that reggie-1 and -2 are associated with fyn kinase and with the glycosylphosphatidyl inositol-anchored proteins Thy-1 and F3 that, when activated by antibody cross-link...

  6. Role of yessotoxin in calcium and cAMP-crosstalks in primary and K-562 human lymphocytes: the effect is mediated by anchor kinase A mitochondrial proteins.

    Science.gov (United States)

    Tobío, Araceli; Fernández-Araujo, Andrea; Alfonso, Amparo; Botana, Luis M

    2012-12-01

    line. Cytosolic expression of A-kinase anchor proteins (AKAPs), the proteins which integrates phosphodiesterases (PDEs) and PKA to the mitochondria, was determined in both cell models. On the one hand, in human fresh lymphocytes, YTX increases AKAP149 cytosolic expression. This fact is accompanied with a decrease in cAMP levels, and therefore PDEs activation, which finally leads to cell survival. On the other hand, in tumoural lymphocytes, YTX has an opposite effect since decreases AKAP149 cytosolic expression and increase cAMP levels which leads to cell death. This is the first time that YTX and mitochondrial AKAPs proteins relationship is characterised.

  7. The molybdenum cofactor biosynthesis complex interacts with actin filaments via molybdenum insertase Cnx1 as anchor protein in Arabidopsis thaliana.

    Science.gov (United States)

    Kaufholdt, David; Baillie, Christin-Kirsty; Bikker, Rolf; Burkart, Valentin; Dudek, Christian-Alexander; von Pein, Linn; Rothkegel, Martin; Mendel, Ralf R; Hänsch, Robert

    2016-03-01

    The pterin based molybdenum cofactor (Moco) plays an essential role in almost all organisms. Its biosynthesis is catalysed by six enzymes in a conserved four step reaction pathway. The last three steps are located in the cytoplasm, where a multimeric protein complex is formed to protect the intermediates from degradation. Bimolecular fluorescence complementation was used to test for cytoskeleton association of the Moco biosynthesis enzymes with actin filaments and microtubules using known cytoskeleton associated proteins, thus permitting non-invasive in vivo studies. Coding sequences of binding proteins were cloned via the GATEWAY system. No Moco biosynthesis enzyme showed any interaction with microtubules. However, alone the two domain protein Cnx1 exhibited interaction with actin filaments mediated by both domains with the Cnx1G domain displaying a stronger interaction. Cnx6 showed actin association only if unlabelled Cnx1 was co-expressed in comparable amounts. So Cnx1 is likely to be the anchor protein for the whole biosynthesis complex on actin filaments. A stabilization of the whole Moco biosynthesis complex on the cytoskeleton might be crucial. In addition a micro-compartmentation might either allow a localisation near the mitochondrial ATM3 exporter providing the first Moco intermediate or near one of the three molybdate transporters enabling efficient molybdate incorporation.

  8. Cell Surface Display and Characterization of Rhizopus oryzae Lipase in Pichia pastoris Using Sed1p as an Anchor Protein.

    Science.gov (United States)

    Li, Wenqian; Shi, Hao; Ding, Huaihai; Wang, Liangliang; Zhang, Yu; Li, Xun; Wang, Fei

    2015-07-01

    It has been investigated to conduct the surface displaying of lipase from Rhizopus oryzae onto the cells of Pichia pastoris yeast using Sed1p as an anchor protein. A yeast cell surface display plasmid pPICZαA-rol-histag-sed1p was constructed by fusing rol and sed1p gene fragments into the plasmid pPICZαA, followed by introducing recombinant plasmid into P. pastoris cells. Surface display levels were monitored by Western Blot and immunofluorescence microscopy. The activity of displaying lipase obtained from recombinant mutS reached at 60 U/g-dry cell. In addition, the displaying lipase was stable in broad ranges of temperatures and pH, with the optimum temperature at 40 °C and pH 7.5. These results indicate that the P. pastoris displaying lipase may have potential in whole-cell biocatalyst.

  9. Anchoring secreted proteins in endoplasmic reticulum by plant oleosin: the example of vitamin B12 cellular sequestration by transcobalamin.

    Directory of Open Access Journals (Sweden)

    Laurent Pons

    Full Text Available BACKGROUND: Oleosin is a plant protein localized to lipid droplets and endoplasmic reticulum of plant cells. Our idea was to use it to target functional secretory proteins of interest to the cytosolic side of the endoplasmic reticulum of mammalian cells, through expressing oleosin-containing chimeras. We have designed this approach to create cellular models deficient in vitamin B12 (cobalamin because of the known problematics associated to the obtainment of effective vitamin B12 deficient cell models. This was achieved by the overexpression of transcobalamin inside cells through anchoring to oleosin. METHODOLOGY: chimera gene constructs including transcobalamin-oleosin (TC-O, green fluorescent protein-transcobalamin-oleosin (GFP-TC-O and oleosin-transcobalamin (O-TC were inserted into pAcSG2 and pCDNA3 vectors for expression in sf9 insect cells, Caco2 (colon carcinoma, NIE-115 (mouse neuroblastoma, HEK (human embryonic kidney, COS-7 (Green Monkey SV40-transfected kidney fibroblasts and CHO (Chinese hamster ovary cells. The subcellular localization, the changes in vitamin B12 binding activity and the metabolic consequences were investigated in both Caco2 and NIE-115 cells. PRINCIPAL FINDINGS: vitamin B12 binding was dramatically higher in TC-O than that in O-TC and wild type (WT. The expression of GFP-TC-O was observed in all cell lines and found to be co-localized with an ER-targeted red fluorescent protein and calreticulin of the endoplasmic reticulum in Caco2 and COS-7 cells. The overexpression of TC-O led to B12 deficiency, evidenced by impaired conversion of cyano-cobalamin to ado-cobalamin and methyl-cobalamin, decreased methionine synthase activity and reduced S-adenosyl methionine to S-adenosyl homocysteine ratio, as well as increases in homocysteine and methylmalonic acid concentration. CONCLUSIONS/SIGNIFICANCE: the heterologous expression of TC-O in mammalian cells can be used as an effective strategy for investigating the cellular

  10. A tail-anchored myotonic dystrophy protein kinase isoform induces perinuclear clustering of mitochondria, autophagy, and apoptosis.

    Directory of Open Access Journals (Sweden)

    Ralph J A Oude Ophuis

    Full Text Available BACKGROUND: Studies on the myotonic dystrophy protein kinase (DMPK gene and gene products have thus far mainly concentrated on the fate of length mutation in the (CTGn repeat at the DNA level and consequences of repeat expansion at the RNA level in DM1 patients and disease models. Surprisingly little is known about the function of DMPK protein products. METHODOLOGY/PRINCIPAL FINDINGS: We demonstrate here that transient expression of one major protein product of the human gene, the hDMPK A isoform with a long tail anchor, results in mitochondrial fragmentation and clustering in the perinuclear region. Clustering occurred in a variety of cell types and was enhanced by an intact tubulin cytoskeleton. In addition to morphomechanical changes, hDMPK A expression induces physiological changes like loss of mitochondrial membrane potential, increased autophagy activity, and leakage of cytochrome c from the mitochondrial intermembrane space accompanied by apoptosis. Truncation analysis using YFP-hDMPK A fusion constructs revealed that the protein's tail domain was necessary and sufficient to evoke mitochondrial clustering behavior. CONCLUSION/SIGNIFICANCE: Our data suggest that the expression level of the DMPK A isoform needs to be tightly controlled in cells where the hDMPK gene is expressed. We speculate that aberrant splice isoform expression might be a codetermining factor in manifestation of specific DM1 features in patients.

  11. Anchoring mechanisms of membrane-associated M13 major coat protein

    NARCIS (Netherlands)

    Stopar, D.; Spruijt, R.B.; Hemminga, M.A.

    2006-01-01

    Bacteriophage M13 major coat protein is extensively used as a biophysical, biochemical, and molecular biology reference system for studying membrane proteins. The protein has several elements that control its position and orientation in a lipid bilayer. The N-terminus is dominated by the presence of

  12. Structure and dynamics of the conserved protein GPI anchor core inserted into detergent micelles.

    Science.gov (United States)

    Chevalier, Franck; Lopez-Prados, Javier; Groves, Patrick; Perez, Serge; Martín-Lomas, Manuel; Nieto, Pedro M

    2006-10-01

    A suitable approach which combines nuclear magnetic resonance (NMR) spectroscopy and molecular dynamics (MD) simulations have been used to study the structure and the dynamics of the glycosylphosphatidylinositol (GPI) anchor Manalphal-2Manalpha1-6Manalphal -4GlcNalpha1-6myo-inositol-1-OPO(3)-sn-1,2-dimyristoylglycerol (1) incorporated into dodecylphosphatidylcholine (DPC) micelles. The results have been compared to those previously obtained for the products obtainable from (1) after phospholipase cleavage, in aqueous solution. Relaxation and diffusion NMR experiments were used to establish the formation of stable aggregates and the insertion of (1) into the micelles. MD calculations were performed including explicit water, sodium and chloride ions and using the Particle Mesh Ewald approach for the evaluation of the electrostatic energy term. The MD predicted three dimensional structure and dynamics were substantiated by nuclear overhauser effect (NOE) measurements and relaxation data. The pseudopentasaccharide structure, which was not affected by incorporation of (1) into the micelle, showed a complex dynamic behaviour with a faster relative motion at the terminal mannopyranose unit and decreased mobility close to the micelle. This motion may be better described as an oscillation relative to the membrane rather than a folding event.

  13. Plasmodium falciparum Plasmodium helical interspersed subtelomeric proteins contribute to cytoadherence and anchor P. falciparum erythrocyte membrane protein 1 to the host cell cytoskeleton

    DEFF Research Database (Denmark)

    Oberli, Alexander; Zurbrügg, Laura; Rusch, Sebastian;

    2016-01-01

    Adherence of Plasmodium falciparum-infected erythrocytes to host endothelium is conferred through the parasite-derived virulence factor P. falciparum erythrocyte membrane protein 1 (PfEMP1), the major contributor to malaria severity. PfEMP1 located at knob structures on the erythrocyte surface...... is anchored to the cytoskeleton, and the Plasmodium helical interspersed subtelomeric (PHIST) gene family plays a role in many host cell modifications including binding the intracellular domain of PfEMP1. Here, we show that conditional reduction of the PHIST protein PFE1605w strongly reduces adhesion...... of infected erythrocytes to the endothelial receptor CD36. Adhesion to other endothelial receptors was less affected or even unaltered by PFE1605w depletion, suggesting that PHIST proteins might be optimized for subsets of PfEMP1 variants. PFE1605w does not play a role in PfEMP1 transport, but it directly...

  14. C-terminal sequences of hsp70 and hsp90 as non-specific anchors for tetratricopeptide repeat (TPR) proteins.

    Science.gov (United States)

    Ramsey, Andrew J; Russell, Lance C; Chinkers, Michael

    2009-10-12

    Steroid-hormone-receptor maturation is a multi-step process that involves several TPR (tetratricopeptide repeat) proteins that bind to the maturation complex via the C-termini of hsp70 (heat-shock protein 70) and hsp90 (heat-shock protein 90). We produced a random T7 peptide library to investigate the roles played by the C-termini of the two heat-shock proteins in the TPR-hsp interactions. Surprisingly, phages with the MEEVD sequence, found at the C-terminus of hsp90, were not recovered from our biopanning experiments. However, two groups of phages were isolated that bound relatively tightly to HsPP5 (Homo sapiens protein phosphatase 5) TPR. Multiple copies of phages with a C-terminal sequence of LFG were isolated. These phages bound specifically to the TPR domain of HsPP5, although mutation studies produced no evidence that they bound to the domain's hsp90-binding groove. However, the most abundant family obtained in the initial screen had an aspartate residue at the C-terminus. Two members of this family with a C-terminal sequence of VD appeared to bind with approximately the same affinity as the hsp90 C-12 control. A second generation pseudo-random phage library produced a large number of phages with an LD C-terminus. These sequences acted as hsp70 analogues and had relatively low affinities for hsp90-specific TPR domains. Unfortunately, we failed to identify residues near hsp90's C-terminus that impart binding specificity to individual hsp90-TPR interactions. The results suggest that the C-terminal sequences of hsp70 and hsp90 act primarily as non-specific anchors for TPR proteins.

  15. A cholesterol recognition amino acid consensus domain in GP64 fusion protein facilitates anchoring of baculovirus to mammalian cells.

    Science.gov (United States)

    Luz-Madrigal, Agustin; Asanov, Alexander; Camacho-Zarco, Aldo R; Sampieri, Alicia; Vaca, Luis

    2013-11-01

    Baculoviridae is a large family of double-stranded DNA viruses that selectively infect insects. Autographa californica multiple nucleopolyhedrovirus (AcMNPV) is the best-studied baculovirus from the family. Many studies over the last several years have shown that AcMNPV can enter a wide variety of mammalian cells and deliver genetic material for foreign gene expression. While most animal viruses studied so far have developed sophisticated mechanisms to selectively infect specific cells and tissues in an organism, AcMNPV can penetrate and deliver foreign genes into most cells studied to this date. The details about the mechanisms of internalization have been partially described. In the present study, we have identified a cholesterol recognition amino acid consensus (CRAC) domain present in the AcMNPV envelope fusion protein GP64. We demonstrated the association of a CRAC domain with cholesterol, which is important to facilitate the anchoring of the virus at the mammalian cell membrane. Furthermore, this initial anchoring favors AcMNPV endocytosis via a dynamin- and clathrin-dependent mechanism. Under these conditions, efficient baculovirus-driven gene expression is obtained. In contrast, when cholesterol is reduced from the plasma membrane, AcMNPV enters the cell via a dynamin- and clathrin-independent mechanism. The result of using this alternative internalization pathway is a reduced level of baculovirus-driven gene expression. This study is the first to document the importance of a novel CRAC domain in GP64 and its role in modulating gene delivery in AcMNPV.

  16. A Cholesterol Recognition Amino Acid Consensus Domain in GP64 Fusion Protein Facilitates Anchoring of Baculovirus to Mammalian Cells

    Science.gov (United States)

    Luz-Madrigal, Agustin; Asanov, Alexander; Camacho-Zarco, Aldo R.; Sampieri, Alicia

    2013-01-01

    Baculoviridae is a large family of double-stranded DNA viruses that selectively infect insects. Autographa californica multiple nucleopolyhedrovirus (AcMNPV) is the best-studied baculovirus from the family. Many studies over the last several years have shown that AcMNPV can enter a wide variety of mammalian cells and deliver genetic material for foreign gene expression. While most animal viruses studied so far have developed sophisticated mechanisms to selectively infect specific cells and tissues in an organism, AcMNPV can penetrate and deliver foreign genes into most cells studied to this date. The details about the mechanisms of internalization have been partially described. In the present study, we have identified a cholesterol recognition amino acid consensus (CRAC) domain present in the AcMNPV envelope fusion protein GP64. We demonstrated the association of a CRAC domain with cholesterol, which is important to facilitate the anchoring of the virus at the mammalian cell membrane. Furthermore, this initial anchoring favors AcMNPV endocytosis via a dynamin- and clathrin-dependent mechanism. Under these conditions, efficient baculovirus-driven gene expression is obtained. In contrast, when cholesterol is reduced from the plasma membrane, AcMNPV enters the cell via a dynamin- and clathrin-independent mechanism. The result of using this alternative internalization pathway is a reduced level of baculovirus-driven gene expression. This study is the first to document the importance of a novel CRAC domain in GP64 and its role in modulating gene delivery in AcMNPV. PMID:23986592

  17. Hormonal activation of a kinase cascade localized at the mitochondria is required for StAR protein activity.

    Science.gov (United States)

    Poderoso, Cecilia; Maloberti, Paula; Duarte, Alejandra; Neuman, Isabel; Paz, Cristina; Cornejo Maciel, Fabiana; Podesta, Ernesto J

    2009-03-01

    It is known that ERK1/2 and MEK1/2 participate in the regulation of Star gene transcription. However, their role in StAR protein post-transcriptional regulation is not described yet. In this study we analyzed the relationship between the MAPK cascade and StAR protein phosphorylation and function. We have demonstrated that (a) steroidogenesis in MA-10 Leydig cells depends on the specific of ERK1/2 activation at the mitochondria; (b) ERK1/2 phosphorylation is driven by mitochondrial PKA and constitutive MEK1/2 in this organelle; (c) active ERK1/2 interacts with StAR protein, leads to StAR protein phosphorylation at Ser(232) only in the presence of cholesterol; (d) directed mutagenesis of Ser(232) (S232A) inhibited in vitro StAR protein phosphorylation by ERK1; (e) transient transfection of MA-10 cells with StAR S232A cDNA markedly reduced the yield of progesterone production. We show that StAR protein is a substrate of ERK1/2, and that mitochondrial ERK1/2 is part of a multimeric complex that regulates cholesterol transport.

  18. Increased Expression of the Tail-Anchored Membrane Protein SLMAP in Adipose Tissue from Type 2 Tally Ho Diabetic Mice

    Directory of Open Access Journals (Sweden)

    Xiaoliang Chen

    2011-01-01

    Full Text Available The tail-anchored membrane protein, sarcolemmal membrane associated protein (SLMAP is encoded to a single gene that maps to the chromosome 3p14 region and has also been reported in certain diabetic populations. Our previous studies with db/db mice shown that a deregulation of SLMAP expression plays an important role in type 2 diabetes. Male Tally Ho mice were bred to present with either normoglycemia (NG or hyperglycemia (HG. Abdominal adipose tissue from male Tally Ho mice of the HG group was found to have a significantly lower expression of the membrane associated glucose transporter-4 (GLUT-4 and higher expression of SLMAP compared to tissue from NG mice. There were 3 isoforms expressed in the abdominal adipose tissue, but only 45?kDa isoform of SLMAP was associated with the GLUT-4 revealed by immunoprecipitation data. Knock down studies using SLMAP siRNA with adipocytes resulted in a significant reduction in SLMAP and a decrease in glucose uptake. Thus, SLMAP may be an important regulator of glucose uptake or involved in GLUT-4 fusion/translocation into the plasma membrane of mouse abdominal adipose tissue and changes in SLMAP expression are linked to hyperglycemia and diabetes.

  19. Anchor Modeling

    Science.gov (United States)

    Regardt, Olle; Rönnbäck, Lars; Bergholtz, Maria; Johannesson, Paul; Wohed, Petia

    Maintaining and evolving data warehouses is a complex, error prone, and time consuming activity. The main reason for this state of affairs is that the environment of a data warehouse is in constant change, while the warehouse itself needs to provide a stable and consistent interface to information spanning extended periods of time. In this paper, we propose a modeling technique for data warehousing, called anchor modeling, that offers non-destructive extensibility mechanisms, thereby enabling robust and flexible management of changes in source systems. A key benefit of anchor modeling is that changes in a data warehouse environment only require extensions, not modifications, to the data warehouse. This ensures that existing data warehouse applications will remain unaffected by the evolution of the data warehouse, i.e. existing views and functions will not have to be modified as a result of changes in the warehouse model.

  20. Bst1 is required for Candida albicans infecting host via facilitating cell wall anchorage of Glycosylphosphatidyl inositol anchored proteins

    Science.gov (United States)

    Liu, Wei; Zou, Zui; Huang, Xin; Shen, Hui; He, Li Juan; Chen, Si Min; Li, Li Ping; Yan, Lan; Zhang, Shi Qun; Zhang, Jun Dong; Xu, Zheng; Xu, Guo Tong; An, Mao Mao; Jiang, Yuan Ying

    2016-01-01

    Glycosylphosphatidyl inositol anchored proteins (GPI-APs) on fungal cell wall are essential for invasive infections. While the function of inositol deacylation of GPI-APs in mammalian cells has been previously characterized the impact of inositol deacylation in fungi and implications to host infection remains largely unexplored. Herein we describe our identification of BST1, an inositol deacylase of GPI-Aps in Candida albicans, was critical for GPI-APs cell wall attachment and host infection. BST1-deficient C. albicans (bst1Δ/Δ) was associated with severely impaired cell wall anchorage of GPI-APs and subsequen unmasked β-(1,3)-glucan. Consistent with the aberrant cell wall structures, bst1Δ/Δ strain did not display an invasive ability and could be recognized more efficiently by host immune systems. Moreover, BST1 null mutants or those expressing Bst1 variants did not display inositol deacylation activity and exhibited severely attenuated virulence and reduced organic colonization in a murine systemic candidiasis model. Thus, Bst1 can facilitate cell wall anchorage of GPI-APs in C. albicans by inositol deacylation, and is critical for host invasion and immune escape. PMID:27708385

  1. Analysis of novel iron-regulated, surface-anchored hemin-binding proteins in Corynebacterium diphtheriae.

    Science.gov (United States)

    Allen, Courtni E; Burgos, Jonathan M; Schmitt, Michael P

    2013-06-01

    Corynebacterium diphtheriae utilizes hemin and hemoglobin (Hb) as iron sources during growth in iron-depleted environments, and recent studies have shown that the surface-exposed HtaA protein binds both hemin and Hb and also contributes to the utilization of hemin iron. Conserved (CR) domains within HtaA and in the associated hemin-binding protein, HtaB, are required for the ability to bind hemin and Hb. In this study, we identified and characterized two novel genetic loci in C. diphtheriae that encode factors that bind hemin and Hb. Both genetic systems contain two-gene operons that are transcriptionally regulated by DtxR and iron. The gene products of these operons are ChtA-ChtB and ChtC-CirA (previously DIP0522-DIP0523). The chtA and chtB genes are carried on a putative composite transposon associated with C. diphtheriae isolates that dominated the diphtheria outbreak in the former Soviet Union in the 1990s. ChtA and ChtC each contain a single N-terminal CR domain and exhibit significant sequence similarity to each other but only limited similarity with HtaA. The chtB and htaB gene products exhibited a high level of sequence similarity throughout their sequences, and both proteins contain a single CR domain. Whole-cell binding studies as well as protease analysis indicated that all four of the proteins encoded by these two operons are surface exposed, which is consistent with the presence of a transmembrane segment in their C-terminal regions. ChtA, ChtB, and ChtC are able to bind hemin and Hb, with ChtA showing the highest affinity. Site-directed mutagenesis showed that specific tyrosine residues within the ChtA CR domain were critical for hemin and Hb binding. Hemin iron utilization assays using various C. diphtheriae mutants indicate that deletion of the chtA-chtB region and the chtC gene has no affect on the ability of C. diphtheriae to use hemin or Hb as iron sources; however, a chtB htaB double mutant exhibits a significant decrease in hemin iron use

  2. FlaF Is a β-Sandwich Protein that Anchors the Archaellum in the Archaeal Cell Envelope by Binding the S-Layer Protein.

    Science.gov (United States)

    Banerjee, Ankan; Tsai, Chi-Lin; Chaudhury, Paushali; Tripp, Patrick; Arvai, Andrew S; Ishida, Justin P; Tainer, John A; Albers, Sonja-Verena

    2015-05-05

    Archaea employ the archaellum, a type IV pilus-like nanomachine, for swimming motility. In the crenarchaeon Sulfolobus acidocaldarius, the archaellum consists of seven proteins: FlaB/X/G/F/H/I/J. FlaF is conserved and essential for archaellum assembly but no FlaF structures exist. Here, we truncated the FlaF N terminus and solved 1.5-Å and 1.65-Å resolution crystal structures of this monotopic membrane protein. Structures revealed an N-terminal α-helix and an eight-strand β-sandwich, immunoglobulin-like fold with striking similarity to S-layer proteins. Crystal structures, X-ray scattering, and mutational analyses suggest dimer assembly is needed for in vivo function. The sole cell envelope component of S. acidocaldarius is a paracrystalline S-layer, and FlaF specifically bound to S-layer protein, suggesting that its interaction domain is located in the pseudoperiplasm with its N-terminal helix in the membrane. From these data, FlaF may act as the previously unknown archaellum stator protein that anchors the rotating archaellum to the archaeal cell envelope.

  3. The late endosomal HOPS complex anchors active G-protein signaling essential for pathogenesis in magnaporthe oryzae.

    Directory of Open Access Journals (Sweden)

    Ravikrishna Ramanujam

    Full Text Available In Magnaporthe oryzae, the causal ascomycete of the devastating rice blast disease, the conidial germ tube tip must sense and respond to a wide array of requisite cues from the host in order to switch from polarized to isotropic growth, ultimately forming the dome-shaped infection cell known as the appressorium. Although the role for G-protein mediated Cyclic AMP signaling in appressorium formation was first identified almost two decades ago, little is known about the spatio-temporal dynamics of the cascade and how the signal is transmitted through the intracellular network during cell growth and morphogenesis. In this study, we demonstrate that the late endosomal compartments, comprising of a PI3P-rich (Phosphatidylinositol 3-phosphate highly dynamic tubulo-vesicular network, scaffold active MagA/GαS, Rgs1 (a GAP for MagA, Adenylate cyclase and Pth11 (a non-canonical GPCR in the likely absence of AKAP-like anchors during early pathogenic development in M. oryzae. Loss of HOPS component Vps39 and consequently the late endosomal function caused a disruption of adenylate cyclase localization, cAMP signaling and appressorium formation. Remarkably, exogenous cAMP rescued the appressorium formation defects associated with VPS39 deletion in M. oryzae. We propose that sequestration of key G-protein signaling components on dynamic late endosomes and/or endolysosomes, provides an effective molecular means to compartmentalize and control the spatio-temporal activation and rapid downregulation (likely via vacuolar degradation of cAMP signaling amidst changing cellular geometry during pathogenic development in M. oryzae.

  4. Comprehensive behavioral analysis of voltage-gated calcium channel beta-anchoring and -regulatory protein knockout mice

    Directory of Open Access Journals (Sweden)

    Akito eNakao

    2015-06-01

    Full Text Available Calcium (Ca2+ influx through voltage-gated Ca2+ channels (VGCCs induces numerous intracellular events such as neuronal excitability, neurotransmitter release, synaptic plasticity, and gene regulation. It has been shown that genes related to Ca2+ signaling, such as the CACNA1C, CACNB2, and CACNA1I genes that encode VGCC subunits, are associated with schizophrenia and other psychiatric disorders. Recently, VGCC beta-anchoring and -regulatory protein (BARP was identified as a novel regulator of VGCC activity via the interaction of VGCC β subunits. To examine the role of the BARP in higher brain functions, we generated BARP knockout (KO mice and conducted a comprehensive battery of behavioral tests. BARP KO mice exhibited greatly reduced locomotor activity, as evidenced by decreased vertical activity, stereotypic counts in the open field test, and activity level in the home cage, and longer latency to complete a session in spontaneous T-maze alteration test, which reached study-wide significance. Acoustic startle response was also reduced in the mutants. Interestingly, they showed multiple behavioral phenotypes that are seemingly opposite to those seen in the mouse models of schizophrenia and its related disorders, including increased working memory, flexibility, prepulse inhibition, and social interaction, and decreased locomotor activity, though many of these phenotypes are statistically weak and require further replications. These results demonstrate that BARP is involved in the regulation of locomotor activity and, possibly, emotionality. The possibility was also suggested that BARP KO mice may serve as a unique tool for investigating the pathogenesis/pathophysiology of schizophrenia and related disorders. Further evaluation of the molecular and physiological phenotypes of the mutant mice would provide new insights into the role of BARP in higher brain functions.

  5. Nanobiotechnologic approach to a promising vaccine prototype for immunisation against leishmaniasis: a fast and effective method to incorporate GPI-anchored proteins of Leishmania amazonensis into liposomes.

    Science.gov (United States)

    Colhone, Marcelle Carolina; Silva-Jardim, Izaltina; Stabeli, Rodrigo Guerino; Ciancaglini, Pietro

    2015-01-01

    Liposomes are known to be a potent adjuvant for a wide range of antigens, as well as appropriate antigen carriers for antibody generation response in vivo. In addition, liposomes are effective vehicles for peptides and proteins, thus enhancing their immunogenicity. Considering these properties of liposomes and the antigenicity of the Leishmania membrane proteins, we evaluated if liposomes carrying glycosylphosphatidylinositol (GPI)-anchored proteins of Leishmania amazonensis promastigotes could induce protective immunity in BALB/c mice. To assay protective immunity, BALB/c mice were intraperitoneally injected with liposomes, GPI-protein extract (EPSGPI) as well as with the proteoliposomes carrying GPI-proteins. Mice inoculated with EPSGPI and total protein present in constitutive proteoliposomes displayed a post-infection protection of about 70% and 90%, respectively. The liposomes are able to work as adjuvant in the EPSGPI protection. These systems seem to be a promising vaccine prototype for immunisation against leishmaniasis.

  6. An improved system for the surface immobilisation of proteins on Bacillus thuringiensis vegetative cells and spores through a new spore cortex-lytic enzyme anchor.

    Science.gov (United States)

    Shao, Xiaohu; Ni, Hong; Lu, Ting; Jiang, Mengtian; Li, Hua; Huang, Xinfeng; Li, Lin

    2012-02-15

    An improved surface-immobilisation system was engineered to target heterologous proteins onto vegetative cells and spores of Bacillus thuringiensis plasmid-free recipient strain BMB171. The sporulation-dependent spore cortex-lytic enzyme from B. thuringiensis YBT-1520, SceA, was expressed in vegetative cells and used as the surface anchoring motif. Green fluorescent protein (GFP) and a Bacillus endo-β-1,3-1,4-glucanase (BglS) were used as the fusion partners to test the binding efficiency and the functional activities of immobilised surface proteins. The surface localisation of the SceA-GFP fusion protein on vegetative cells and spores was confirmed by Western blot, immunofluorescence microscopy and flow cytometry. The GFP fluorescence intensity from both vegetative cells and spores was measured and compared to a previously characterised surface display system using a peptidoglycan hydrolase anchor (Mbg). Results demonstrated comparable efficiency of SceA- and Mbg-mediated immobilisation on vegetative cells but a more efficient immobilisation on spores using the SceA anchor, suggesting SceA has greater potential for spore-based applications. The SceA protein was then applied to target BglS onto vegetative cells and spores, and the surface immobilisation was verified by the substantial whole-cell enzymatic activity and enhanced whole-spore enzymatic activity compared to vegetative cells. A dually active B. thuringiensis vegetative cell and spore display system could prove especially valuable for the development of regenerable and heat-stable biocatalysts that function under adverse environmental conditions, for example, an effective feed additive for improved digestion and nutrient absorption by livestock.

  7. Recording intracellular molecular events from the outside: glycosylphosphatidylinositol-anchored avidin as a reporter protein for in vivo imaging.

    NARCIS (Netherlands)

    Lehmann, S.A.; Garayoa, E.G.; Blanc, A.; Keist, R.; Schibli, R.; Rudin, M.

    2011-01-01

    With the emergence of multimodal imaging strategies, genetically encoded reporters that can be flexibly combined with any imaging modality become highly attractive. Here we describe the use of glycosylphosphatidylinositol (GPI)-anchored avidin, an avidin moiety targeted to the extracellular side of

  8. GPI-80, a beta2 integrin associated glycosylphosphatidylinositol-anchored protein, concentrates on pseudopodia without association with beta2 integrin during neutrophil migration.

    Science.gov (United States)

    Yoshitake, Hiroshi; Takeda, Yuji; Nitto, Takeaki; Sendo, Fujiro; Araki, Yoshihiko

    2003-01-01

    Previously, we identified a glycosylphosphatidylinositol (GPI)-anchored protein, designated GPI-80, present on human neutrophils and monocytes. GPI-80 is physically associated with beta2 integrin on the surface of human neutrophils and may be a regulator of neutrophil adherence and migration. However, it is not yet known how GPI-80 regulates cell adhesion and migration. To investigate the physiological role(s) of GPI-80, we examined the topological relationship of GPI-80 and the beta2 integrin subunit (CD18) on resting and migrating human neutrophils by confocal laser microscopy. On resting neutrophils, GPI-80 was evenly distributed on the cell surface and was associated with CD18. On the other hand, during the early phase of migration (5 - 30 minutes), GPI-80 was detected on cell bodies and also on pseudopodia, but CD18 was detected only on cell bodies, where it was associated with GPI-80. In the late phase of migration (60 minutes), GPI-80 was detected only on pseudopodia and its association with CD18 was hardly observed. Furthermore, some of the GPI-80 on pseudopodia of migrating neutrophils during the late phase was associated with urokinase-type plasminogen activator receptor (uPAR), a regulator of beta2 integrin-dependent adherence and migration. The distribution of GPI-80 on cell surfaces is similar to that of uPAR. These observations suggest that GPI-80 belongs to the beta2 integrin-associated GPI-anchored protein family, which has regulatory activity in cell adherence.

  9. Gravin orchestrates protein kinase A and β2-adrenergic receptor signaling critical for synaptic plasticity and memory

    NARCIS (Netherlands)

    Havekes, Robbert; Canton, David A; Park, Alan J; Huang, Ted; Nie, Ting; Day, Jonathan P; Guercio, Leonardo A; Grimes, Quinn; Luczak, Vincent; Gelman, Irwin H; Baillie, George S; Scott, John D; Abel, Ted

    2012-01-01

    A kinase-anchoring proteins (AKAPs) organize compartmentalized pools of protein kinase A (PKA) to enable localized signaling events within neurons. However, it is unclear which of the many expressed AKAPs in neurons target PKA to signaling complexes important for long-lasting forms of synaptic plast

  10. Plasmodium falciparum Plasmodium helical interspersed subtelomeric proteins contribute to cytoadherence and anchor P. falciparum erythrocyte membrane protein 1 to the host cell cytoskeleton

    DEFF Research Database (Denmark)

    Oberli, Alexander; Zurbrügg, Laura; Rusch, Sebastian

    2016-01-01

    is anchored to the cytoskeleton, and the Plasmodium helical interspersed subtelomeric (PHIST) gene family plays a role in many host cell modifications including binding the intracellular domain of PfEMP1. Here, we show that conditional reduction of the PHIST protein PFE1605w strongly reduces adhesion...... of infected erythrocytes to the endothelial receptor CD36. Adhesion to other endothelial receptors was less affected or even unaltered by PFE1605w depletion, suggesting that PHIST proteins might be optimized for subsets of PfEMP1 variants. PFE1605w does not play a role in PfEMP1 transport, but it directly...

  11. Efficient cell surface display of Lip2 lipase using C-domains of glycosylphosphatidylinositol-anchored cell wall proteins of Yarrowia lipolytica.

    Science.gov (United States)

    Yuzbasheva, Evgeniya Y; Yuzbashev, Tigran V; Laptev, Ivan A; Konstantinova, Tatiana K; Sineoky, Sergey P

    2011-08-01

    The cell surface display of enzymes is of great interest because of its simplified purification stage and the possibility for recycling in industrial processes. In this study, we have focused on the cell wall immobilization of Yarrowia lipolytica Lip2 protein--an enzyme that has a wide technological application. By genome analysis of Y. lipolytica in addition to already characterized Ylcwp1, we identified five putative open reading frames encoding glycosylphosphatidylinositol-anchored proteins. Lip2 translation fusion with the carboxyl termini of these proteins revealed that all proteins were capable of immobilizing lipase in active form on the cell surface. The highest level of cell-bound lipase activity was achieved using C-domains encoded by YlCWP1, YlCWP3 (YALI0D27214g) and YlCWP6 (YALI0F18282g) comprising 16,173 ± 1,800, 18,785 ± 1,130 and 17,700 ± 2,101 U/g dry cells, respectively. To the best of our knowledge, these results significantly exceed the highest cell-bound lipase activity previously reported for engineered Saccharomyces cerevisiae and Pichia pastoris strains. Furthermore, the lyophilized biomass retained the activity and was robust to collecting/resuspending procedures. Nevertheless, in most cases, a substantial amount of lipase activity was also found in the growth medium. Further work will be necessary to better understand the nature of this phenomenon.

  12. Glycosylphosphatidylinositol Anchor Modification Machinery Deficiency Is Responsible for the Formation of Pro-Prion Protein (PrP) in BxPC-3 Protein and Increases Cancer Cell Motility.

    Science.gov (United States)

    Yang, Liheng; Gao, Zhenxing; Hu, Lipeng; Wu, Guiru; Yang, Xiaowen; Zhang, Lihua; Zhu, Ying; Wong, Boon-Seng; Xin, Wei; Sy, Man-Sun; Li, Chaoyang

    2016-02-19

    The normal cellular prion protein (PrP) is a glycosylphosphatidylinositol (GPI)-anchored cell surface glycoprotein. However, in pancreatic ductal adenocarcinoma cell lines, such as BxPC-3, PrP exists as a pro-PrP retaining its glycosylphosphatidylinositol (GPI) peptide signaling sequence. Here, we report the identification of another pancreatic ductal adenocarcinoma cell line, AsPC-1, which expresses a mature GPI-anchored PrP. Comparison of the 24 genes involved in the GPI anchor modification pathway between AsPC-1 and BxPC-3 revealed 15 of the 24 genes, including PGAP1 and PIG-F, were down-regulated in the latter cells. We also identified six missense mutations in DPM2, PIG-C, PIG-N, and PIG-P alongside eight silent mutations. When BxPC-3 cells were fused with Chinese hamster ovary (CHO) cells, which lack endogenous PrP, pro-PrP was successfully converted into mature GPI-anchored PrP. Expression of the individual gene, such as PGAP1, PIG-F, or PIG-C, into BxPC-3 cells does not result in phosphoinositide-specific phospholipase C sensitivity of PrP. However, when PIG-F but not PIG-P is expressed in PGAP1-expressing BxPC-3 cells, PrP on the surface of the cells becomes phosphoinositide-specific phospholipase C-sensitive. Thus, low expression of PIG-F and PGAP1 is the major factor contributing to the accumulation of pro-PrP. More importantly, BxPC-3 cells expressing GPI-anchored PrP migrate much slower than BxPC-3 cells bearing pro-PrP. In addition, GPI-anchored PrP-bearing AsPC-1 cells also migrate slower than pro-PrP bearing BxPC-3 cells, although both cells express filamin A. "Knocking out" PRNP in BxPC-3 cell drastically reduces its migration. Collectively, these results show that multiple gene irregularity in BxPC-3 cells is responsible for the formation of pro-PrP, and binding of pro-PrP to filamin A contributes to enhanced tumor cell motility.

  13. In silicio identification of glycosyl-phosphatidylinositol-anchored plasma-membrane and cell wall proteins of Saccharomyces cerevisiae.

    Science.gov (United States)

    Caro, L H; Tettelin, H; Vossen, J H; Ram, A F; van den Ende, H; Klis, F M

    1997-12-01

    Use of the Von Heijne algorithm allowed the identification of 686 open reading frames (ORFs) in the genome of Saccharomyces cerevisiae that encode proteins with a potential N-terminal signal sequence for entering the secretory pathway. On further analysis, 51 of these proteins contain a potential glycosyl-phosphatidylinositol (GPI)-attachment signal. Seven additional ORFs were found to belong to this group. Upon examination of the possible GPI-attachment sites, it was found that in yeast the most probable amino acids for GPI-attachment as asparagine and glycine. In yeast, GPI-proteins are found at the cell surface, either attached to the plasma-membrane or as an intrinsic part of the cell wall. It was noted that plasma-membrane GPI-proteins possess a dibasic residue motif just before their predicted GPI-attachment site. Based on this, and on homologies between proteins, families of plasma-membrane and cell wall proteins were assigned, revealing 20 potential plasma-membrane and 38 potential cell wall proteins. For members of three plasma-membrane protein families, a function has been described. On the other hand, most of the cell wall proteins seem to be structural components of the wall, responsive to different growth conditions. The GPI-attachment site of yeast slightly differs from mammalian cells. This might be of use in the development of anti-fungal drugs.

  14. Roles of the protruding loop of factor B essential for the localization of lipoproteins (LolB) in the anchoring of bacterial triacylated proteins to the outer membrane.

    Science.gov (United States)

    Hayashi, Yumi; Tsurumizu, Ryoji; Tsukahara, Jun; Takeda, Kazuki; Narita, Shin-ichiro; Mori, Makiko; Miki, Kunio; Tokuda, Hajime

    2014-04-11

    The Lol system comprising five Lol proteins, LolA through LolE, sorts Escherichia coli lipoproteins to outer membranes. The LolCDE complex, an ATP binding cassette transporter in inner membranes, releases outer membrane-specific lipoproteins in an ATP-dependent manner, causing formation of the LolA-lipoprotein complex in the periplasm. LolA transports lipoproteins through the periplasm to LolB on outer membranes. LolB is itself a lipoprotein anchored to outer membranes, although the membrane anchor is functionally dispensable. LolB then localizes lipoproteins to outer membranes through largely unknown mechanisms. The crystal structure of LolB is similar to that of LolA, and it possesses a hydrophobic cavity that accommodates acyl chains of lipoproteins. To elucidate the molecular function of LolB, a periplasmic version of LolB, mLolB, was mutagenized at various conserved residues. Despite the lack of acyl chains, most defective mutants were insoluble. However, a derivative with glutamate in place of leucine 68 was soluble and unable to localize lipoproteins to outer membranes. This leucine is present in a loop protruding from mLolB into an aqueous environment, and no analogous loop is present in LolA. Thus, leucine 68 was replaced with other residues. Replacement by acidic, but not hydrophobic, residues generated for the first time mLolB derivatives that can accept but cannot localize lipoproteins to outer membranes. Moreover, deletion of the leucine with neighboring residues impaired the lipoprotein receptor activity. Based on these observations, the roles of the protruding loop of LolB in the last step of lipoprotein sorting are discussed.

  15. One repeat of the cell wall binding domain is sufficient for anchoring the Lactobacillus acidophilus surface layer protein

    NARCIS (Netherlands)

    Smit, E.; Pouwels, P.H.

    2002-01-01

    The N-terminal repeat (SAC1) of the S-protein of Lactobacillus acidophilus bound efficiently and specifically to cell wall fragments (CWFs) when fused to green fluorescent protein, whereas the C-terminal repeat (SAC2) did not. Treatment of CWFs with hydrofluoric acid, but not phenol, prevented bindi

  16. Hypersensitivity to Suture Anchors

    Directory of Open Access Journals (Sweden)

    Masafumi Goto

    2013-01-01

    Full Text Available Hypersensitivity to suture anchor is extremely rare. Herein, we present a case in which hypersensitivity to suture anchor was strongly suspected. The right rotator cuff of a 50-year-old woman was repaired with a metal suture anchor. Three weeks after the surgery, she developed erythema around her face, trunk, and hands, accompanied by itching. Infection was unlikely because no abnormalities were detected by blood testing or by medical examination. Suspicious of a metallic allergy, a dermatologist performed a patch testing 6 months after the first surgery. The patient had negative reactions to tests for titanium, aluminum, and vanadium, which were the principal components of the suture anchor. The anchor was removed 7 months after the first surgery, and the erythema disappeared immediately. When allergic symptoms occur and persist after the use of a metal anchor, removal should be considered as a treatment option even if the patch test result is negative.

  17. Interaction of modified tail-anchored proteins with liposomes: effect of extensions of hydrophilic segment at the COOH-terminus of holo-cytochromes b₅.

    Science.gov (United States)

    Sakamoto, Yoichi; Miura, Masahiro; Takeuchi, Fusako; Park, Sam-Yong; Tsubaki, Motonari

    2012-03-01

    A group of membrane proteins having a single COOH-terminal hydrophobic domain capable of post-translational insertion into lipid bilayer is known as tail-anchored (TA) proteins. To clarify the insertion mechanism of the TA-domain of human cytochrome b(5) (Hcytb5) into ER membranes, we produced and purified various membrane-bound forms of Hcytb5 with their heme b-bound, in which various truncated forms of NH(2)-terminal bovine opsin sequence were appended at the COOH-terminus of the native form. We analyzed the integration of the TA-domains of these forms onto protein-free liposomes. The integration occurred efficiently even in the presence of a small amount of sodium cholate and, once incorporated, such proteoliposomes were very stable. The mode of the integration was further analyzed by treatment of the proteoliposomes with trypsin either on the extravesicular side or on the luminal side. LC-MS analyses of the trypsin digests obtained from the proteoliposomes indicated that most of the C-terminal hydrophilic segment of the native Hcytb5 were exposed towards the lumen of the vesicles and, further, a significant part of the population of the extended C-terminal hydrophilic segments of the modified Hcytb5 were exposed in the lumen as well, suggesting efficient translocation ability of the TA-domain without any assistance from other protein factors. Present results opened a route for the use of the C-terminal TA-domain as a convenient tool for the transport of proteins as well as short peptides into artificial liposomes.

  18. Anchor Loads on Pipelines

    OpenAIRE

    Wei, Ying

    2015-01-01

    Anchor hooking on a subsea pipeline has been investigated in this thesis. Anchor loads on pipelines is in general a rarely occurring event, however, the severity when it occurs could easily jeopardize the integrity of any pipeline. It is considered as an accidental load in the design of pipelines. Pipeline Loads, limit state criteria and anchor categories are defined by the DNV standards. For pipeline, DNV-OS-F101 (08.2012), Submarine Pipeline Systems is adopted. Offshore standard DNV-RP...

  19. Design and optimization of a phosphopeptide anchor for specific immobilization of a capture protein on zirconium phosphonate modified supports.

    Science.gov (United States)

    Liu, Hao; Queffélec, Clémence; Charlier, Cathy; Defontaine, Alain; Fateh, Amina; Tellier, Charles; Talham, Daniel R; Bujoli, Bruno

    2014-11-25

    The attachment of affinity proteins onto zirconium phosphonate coated glass slides was investigated by fusing a short phosphorylated peptide sequence at one extremity to enable selective bonding to the active surface via the formation of zirconium phosphate coordinate covalent bonds. In a model study, the binding of short peptides containing zero to four phosphorylated serine units and a biotin end-group was assessed by surface plasmon resonance-enhanced ellipsometry (SPREE) as well as in a microarray format using fluorescence detection of AlexaFluor 647-labeled streptavidin. Significant binding to the zirconated surface was only observed in the case of the phosphopeptides, with the best performance, as judged by streptavidin capture, observed for peptides with three or four phosphorylation sites and when spotted at pH 3. When fusing similar phosphopeptide tags to the affinity protein, the presence of four phosphate groups in the tag allows efficient immobilization of the proteins and efficient capture of their target.

  20. Transforming p21 ras protein: flexibility in the major variable region linking the catalytic and membrane-anchoring domains

    DEFF Research Database (Denmark)

    Willumsen, B M; Papageorge, A G; Hubbert, N;

    1985-01-01

    that is required for post-translational processing, membrane localization and transforming activity of the proteins. We have now used the viral oncogene (v-rasH) of Harvey sarcoma virus to study the major variable region by deleting or duplicating parts of the gene. Reducing this region to five amino acids...... or increasing it to 50 amino acids has relatively little effect on the capacity of the gene to induce morphological transformation of NIH 3T3 cells. Assays of GTP binding, GTPase and autophosphorylating activities of such mutant v-rasH-encoded proteins synthesized in bacteria indicated that the sequences...

  1. Anchoring skeletal muscle development and disease: The role of ankyrin repeat domain containing proteins in muscle physiology

    NARCIS (Netherlands)

    J-M. Tee (Jin-Ming); M.P. Peppelenbosch (Maikel)

    2010-01-01

    textabstractThe ankyrin repeat is a protein module with high affinity for other ankyrin repeats based on strong Van der Waals forces. The resulting dimerization is unusually resistant to both mechanical forces and alkanization, making this module exceedingly useful for meeting the extraordinary dema

  2. Functional characterization and localization of a Bacillus subtilis sortase and its substrate and use of this sortase system to covalently anchor a heterologous protein to the B. subtilis cell wall for surface display.

    Science.gov (United States)

    Liew, Pei Xiong; Wang, Christopher L C; Wong, Sui-Lam

    2012-01-01

    Sortases catalyze the covalent anchoring of proteins to the cell surface on Gram-positive bacteria. Bioinformatic analysis suggests the presence of structural genes encoding sortases and their substrates in the Bacillus subtilis genome. In this study, a β-lactamase reporter was fused to the cell wall anchoring domain from a putative sortase substrate, YhcR. Covalent anchoring of this fusion protein to the cell wall was confirmed by using the eight-protease-deficient B. subtilis strain WB800 as the host. Inactivation of yhcS abolished the cell wall anchoring reaction. The amounts of fusion protein anchored to the cell wall were proportional to the levels of YhcS. These data demonstrate that YhcS and YhcR are the sortase and sortase substrate, respectively, in B. subtilis. Furthermore, yhcS is not essential for the survival of B. subtilis under the cultivation condition tested. YhcR fusions were distributed helically in the lateral cell wall. Interestingly, when viewed with an epifluorescence microscope, YhcS also appeared to form short helical arcs. This is the first report to illustrate such distribution of sortases in a rod-shaped bacterium. Models for the spatial distribution of both the sortase and its substrate are discussed. The amount of the reporters displayed on the surface was unambiguously quantified via a unique strategy. Under optimal conditions with the overproduction of YhcS, 47,300 YhcR fusions could be displayed per cell. Displayed reporters were biologically functional and surface accessible. Characterization of the sortase-substrate system allowed the successful development of a YhcR-based covalent surface display system. This system may have various biotechnological applications.

  3. Susceptibility to anchoring effects

    Directory of Open Access Journals (Sweden)

    Todd McElroy

    2007-02-01

    Full Text Available Previous research on anchoring has shown this heuristic to be a very robust psychological phenomenon ubiquitous across many domains of human judgment and decision-making. Despite the prevalence of anchoring effects, researchers have only recently begun to investigate the underlying factors responsible for how and in what ways a person is susceptible to them. This paper examines how one such factor, the Big-Five personality trait of openness-to-experience, influences the effect of previously presented anchors on participants' judgments. Our findings indicate that participants high in openness-to-experience were significantly more influenced by anchoring cues relative to participants low in this trait. These findings were consistent across two different types of anchoring tasks providing convergent evidence for our hypothesis.

  4. BAR domains, amphipathic helices and membrane-anchored proteins use the same mechanism to sense membrane curvature

    DEFF Research Database (Denmark)

    Madsen, Kenneth Lindegaard; Bhatia, V K; Gether, U;

    2010-01-01

    The internal membranes of eukaryotic cells are all twists and bends characterized by high curvature. During recent years it has become clear that specific proteins sustain these curvatures while others simply recognize membrane shape and use it as "molecular information" to organize cellular...... processes in space and time. Here we discuss this new important recognition process termed membrane curvature sensing (MCS). First, we review a new fluorescence-based experimental method that allows characterization of MCS using measurements on single vesicles and compare it to sensing assays that use bulk...... on curved membranes instead of higher affinity as assumed so far. Finally, we integrate these new insights into the debate about which motifs are involved in sensing versus induction of membrane curvature and what role MCS proteins may play in biology....

  5. Curcumin Protects Neurons from Glutamate-Induced Excitotoxicity by Membrane Anchored AKAP79-PKA Interaction Network

    Directory of Open Access Journals (Sweden)

    Kui Chen

    2015-01-01

    Full Text Available Now stimulation of AMPA receptor as well as its downstream pathways is considered as potential central mediators in antidepressant mechanisms. As a signal integrator which binds to AMPA receptor, A-kinase anchoring protein 79-(AKAP79- PKA complex is regarded as a potential drug target to exert neuroprotective effects. A well-tolerated and multitarget drug curcumin has been confirmed to exert antidepressant-like effects. To explore whether AKAP79-PKA complex is involved in curcumin-mediated antiexcitotoxicity, we detected calcium signaling, subcellular location of AKAP79-PKA complex, phosphorylation of glutamate receptor, and ERK and AKT cascades. In this study, we found that curcumin protected neurons from glutamate insult by reducing Ca2+ influx and blocking the translocation of AKAP79 from cytomembrane to cytoplasm. In parallel, curcumin enhanced the phosphorylation of AMPA receptor and its downstream pathways in PKA-dependent manner. If we pretreated cells with PKA anchoring inhibitor Ht31 to disassociate PKA from AKAP79, no neuroprotective effects were observed. In conclusion, our results show that AKAP79-anchored PKA facilitated the signal relay from AMPA receptor to AKT and ERK cascades, which may be crucial for curcumin-mediated antiexcitotoxicity.

  6. Identification and Characterization of a Novel Issatchenkia orientalis GPI-Anchored Protein, IoGas1, Required for Resistance to Low pH and Salt Stress

    Science.gov (United States)

    Matsushika, Akinori; Negi, Kanako; Suzuki, Toshihiro; Goshima, Tetsuya; Hoshino, Tamotsu

    2016-01-01

    The use of yeasts tolerant to acid (low pH) and salt stress is of industrial importance for several bioproduction processes. To identify new candidate genes having potential roles in low-pH tolerance, we screened an expression genomic DNA library of a multiple-stress-tolerant yeast, Issatchenkia orientalis (Pichia kudriavzevii), for clones that allowed Saccharomyces cerevisiae cells to grow under highly acidic conditions (pH 2.0). A genomic DNA clone containing two putative open reading frames was obtained, of which the putative protein-coding gene comprising 1629 bp was retransformed into the host. This transformant grew significantly at pH 2.0, and at pH 2.5 in the presence of 7.5% Na2SO4. The predicted amino acid sequence of this new gene, named I. orientalis GAS1 (IoGAS1), was 60% identical to the S. cerevisiae Gas1 protein, a glycosylphosphatidylinositol-anchored protein essential for maintaining cell wall integrity, and 58–59% identical to Candida albicans Phr1 and Phr2, pH-responsive proteins implicated in cell wall assembly and virulence. Northern hybridization analyses indicated that, as for the C. albicans homologs, IoGAS1 expression was pH-dependent, with expression increasing with decreasing pH (from 4.0 to 2.0) of the medium. These results suggest that IoGAS1 represents a novel pH-regulated system required for the adaptation of I. orientalis to environments of diverse pH. Heterologous expression of IoGAS1 complemented the growth and morphological defects of a S. cerevisiae gas1Δ mutant, demonstrating that IoGAS1 and the corresponding S. cerevisiae gene play similar roles in cell wall biosynthesis. Site-directed mutagenesis experiments revealed that two conserved glutamate residues (E161 and E262) in the IoGas1 protein play a crucial role in yeast morphogenesis and tolerance to low pH and salt stress. Furthermore, overexpression of IoGAS1 in S. cerevisiae remarkably improved the ethanol fermentation ability at pH 2.5, and at pH 2.0 in the presence of

  7. Salmonella Effectors SseF and SseG Interact with Mammalian Protein ACBD3 (GCP60 To Anchor Salmonella-Containing Vacuoles at the Golgi Network

    Directory of Open Access Journals (Sweden)

    Xiu-Jun Yu

    2016-07-01

    Full Text Available Following infection of mammalian cells, Salmonella enterica serovar Typhimurium (S. Typhimurium replicates within membrane-bound compartments known as Salmonella-containing vacuoles (SCVs. The Salmonella pathogenicity island 2 type III secretion system (SPI-2 T3SS translocates approximately 30 different effectors across the vacuolar membrane. SseF and SseG are two such effectors that are required for SCVs to localize close to the Golgi network in infected epithelial cells. In a yeast two-hybrid assay, SseG and an N-terminal variant of SseF interacted directly with mammalian ACBD3, a multifunctional cytosolic Golgi network-associated protein. Knockdown of ACBD3 by small interfering RNA (siRNA reduced epithelial cell Golgi network association of wild-type bacteria, phenocopying the effect of null mutations of sseG or sseF. Binding of SseF to ACBD3 in infected cells required the presence of SseG. A single-amino-acid mutant of SseG and a double-amino-acid mutant of SseF were obtained that did not interact with ACBD3 in Saccharomyces cerevisiae. When either of these was produced together with the corresponding wild-type effector by Salmonella in infected cells, they enabled SCV-Golgi network association and interacted with ACBD3. However, these properties were lost and bacteria displayed an intracellular replication defect when cells were infected with Salmonella carrying both mutant genes. Knockdown of ACBD3 resulted in a replication defect of wild-type bacteria but did not further attenuate the growth defect of a ΔsseFG mutant strain. We propose a model in which interaction between SseF and SseG enables both proteins to bind ACBD3, thereby anchoring SCVs at the Golgi network and facilitating bacterial replication.

  8. PPP1CC2 can form a kinase/phosphatase complex with the testis-specific proteins TSSK1 and TSKS in the mouse testis

    NARCIS (Netherlands)

    G. MacLeod (Graham); P. Shang (Peng); G.T. Booth (Gregory); L.A. Mastropaolo (Lucas); N. Manafpoursakha (Niloufar); A.W. Vogl (A. Wayne); S. Varmuza (Susannah)

    2014-01-01

    textabstractThe mouse protein phosphatase gene Ppp1cc is essential for male fertility, with mutants displaying a failure in spermatogenesis including a widespread loss of post-meiotic germ cells and abnormalities in the mitochondrial sheath. This phenotype is hypothesized to be responsible for the l

  9. High-throughput automated confocal microscopy imaging screen of a kinase-focused library to identify p38 mitogen-activated protein kinase inhibitors using the GE InCell 3000 analyzer.

    Science.gov (United States)

    Trask, O Joseph; Nickischer, Debra; Burton, Audrey; Williams, Rhonda Gates; Kandasamy, Ramani A; Johnston, Patricia A; Johnston, Paul A

    2009-01-01

    The integration of fluorescent microscopy imaging technologies and image analysis into high-content screening (HCS) has been applied throughout the drug discovery pipeline to identify, evaluate, and advance compounds from early lead generation through preclinical candidate selection. In this chapter we describe the development, validation, and implementation of an HCS assay to screen compounds from a kinase-focused small-molecule library to identify inhibitors of the p38 pathway using the GE InCell 3000 automated imaging platform. The assay utilized a genetically modified HeLa cell line stably expressing mitogen-activated, protein-activating protein kinase-2 fused to enhanced green fluorescent protein (MK2-EGFP) and measured the subcellular distribution of the MK2-EGFP as a direct readout of p38 activation. The MK2-EGFP translocation assay performed in 384-well glass bottom microtiter plates exhibited a robust Z-factor of 0.46 and reproducible EC50 and IC50 determinations for activators and inhibitors, respectively. A total of 32,891 compounds were screened in singlicate at 50 microM and 156 were confirmed as inhibitors of p38-mediated MK2-EGFP translocation in follow-up IC50 concentration response curves. Thirty-one compounds exhibited IC50s less than 1 microM, and at least one novel structural class of p38 inhibitor was identified using this HCA/HCS chemical biology screening approach.

  10. High Cell Sensitivity to Helicobacter pylori VacA Toxin Depends on a GPI-anchored Protein and is not Blocked by Inhibition of the Clathrin-mediated Pathway of Endocytosis

    OpenAIRE

    2000-01-01

    Helicobacter pylori vacuolating toxin (VacA) causes vacuolation in a variety of cultured cell lines, sensitivity to VacA differing greatly, however, among the different cell types. We found that the high sensitivity of HEp-2 cells to VacA was impaired by treating the cells with phosphatidylinositol-specific phospholipase C (PI-PLC) which removes glycosylphosphatidylinositol (GPI)-anchored proteins from the cell surface. Incubation of cells with a cholesterol-seques...

  11. Anchor Trial Launch

    Science.gov (United States)

    NCI has launched a multicenter phase III clinical trial called the ANCHOR Study -- Anal Cancer HSIL (High-grade Squamous Intraepithelial Lesion) Outcomes Research Study -- to determine if treatment of HSIL in HIV-infected individuals can prevent anal canc

  12. Freely turning over palmitate in erythrocyte membrane proteins is not responsible for the anchoring of lipid rafts to the spectrin skeleton: a study with bio-orthogonal chemical probes.

    Science.gov (United States)

    Ciana, Annarita; Achilli, Cesare; Hannoush, Rami N; Risso, Angela; Balduini, Cesare; Minetti, Giampaolo

    2013-03-01

    Erythrocyte lipid rafts are anchored to the underlying spectrin membrane skeleton [A. Ciana, C. Achilli, C. Balduini, G. Minetti, On the association of lipid rafts to the spectrin skeleton in human erythrocytes, Biochim. Biophys. Acta 1808 (2011) 183-190]. The nature of this linkage and the molecules involved are poorly understood. The interaction is sensitive to the increase in pH and ionic strength induced by carbonate. Given the role of palmitoylation in modulating the partitioning of certain proteins between various sub-cellular compartments and the plasma membrane, we asked whether palmitoylation of p55, a peripheral protein located at the junctional complex between spectrin-actin-protein 4.1 that anchors the membrane skeleton to the lipid bilayer via the transmembrane protein glycophorin C, could contribute to the anchoring of lipid rafts to the membrane skeleton. We adopted a new, non-radioactive method for studying protein palmitoylation, based on bio-orthogonal chemical analogues of fatty acids, containing an omega-alkynyl group, to metabolically label cell proteins, which are then revealed by a "click chemistry" reaction of the alkynyl moiety with an azide-containing reporter tag. We show that the membrane localization and palmitoylation levels of p55 did not change after carbonate treatment. 2-bromopalmitate and cerulenin, two known palmitoylation inhibitors, completely inhibited p55 palmitoylation, and protein palmitoyl thioesterase-1 (PPT1) reduced it, without affecting the association between lipid rafts and membrane-skeleton, indicating, on the one hand, that p55 palmitoylation is enzymatic, and, on the other, that it is not involved in the modulation of the linkage of lipid rafts to the membrane-skeleton.

  13. Gpr161 anchoring of PKA consolidates GPCR and cAMP signaling.

    Science.gov (United States)

    Bachmann, Verena A; Mayrhofer, Johanna E; Ilouz, Ronit; Tschaikner, Philipp; Raffeiner, Philipp; Röck, Ruth; Courcelles, Mathieu; Apelt, Federico; Lu, Tsan-Wen; Baillie, George S; Thibault, Pierre; Aanstad, Pia; Stelzl, Ulrich; Taylor, Susan S; Stefan, Eduard

    2016-07-12

    Scaffolding proteins organize the information flow from activated G protein-coupled receptors (GPCRs) to intracellular effector cascades both spatially and temporally. By this means, signaling scaffolds, such as A-kinase anchoring proteins (AKAPs), compartmentalize kinase activity and ensure substrate selectivity. Using a phosphoproteomics approach we identified a physical and functional connection between protein kinase A (PKA) and Gpr161 (an orphan GPCR) signaling. We show that Gpr161 functions as a selective high-affinity AKAP for type I PKA regulatory subunits (RI). Using cell-based reporters to map protein-protein interactions, we discovered that RI binds directly and selectively to a hydrophobic protein-protein interaction interface in the cytoplasmic carboxyl-terminal tail of Gpr161. Furthermore, our data demonstrate that a binary complex between Gpr161 and RI promotes the compartmentalization of Gpr161 to the plasma membrane. Moreover, we show that Gpr161, functioning as an AKAP, recruits PKA RI to primary cilia in zebrafish embryos. We also show that Gpr161 is a target of PKA phosphorylation, and that mutation of the PKA phosphorylation site affects ciliary receptor localization. Thus, we propose that Gpr161 is itself an AKAP and that the cAMP-sensing Gpr161:PKA complex acts as cilium-compartmentalized signalosome, a concept that now needs to be considered in the analyzing, interpreting, and pharmaceutical targeting of PKA-associated functions.

  14. Plasmodium falciparum Plasmodium helical interspersed subtelomeric proteins contribute to cytoadherence and anchor P. falciparum erythrocyte membrane protein 1 to the host cell cytoskeleton

    DEFF Research Database (Denmark)

    Oberli, Alexander; Zurbrügg, Laura; Rusch, Sebastian;

    2016-01-01

    Adherence of Plasmodium falciparum-infected erythrocytes to host endothelium is conferred through the parasite-derived virulence factor P. falciparum erythrocyte membrane protein 1 (PfEMP1), the major contributor to malaria severity. PfEMP1 located at knob structures on the erythrocyte surface is...

  15. High Cell Sensitivity to Helicobacter pylori VacA Toxin Depends on a GPI-anchored Protein and is not Blocked by Inhibition of the Clathrin-mediated Pathway of Endocytosis

    Science.gov (United States)

    Ricci, Vittorio; Galmiche, Antoine; Doye, Anne; Necchi, Vittorio; Solcia, Enrico; Boquet, Patrice

    2000-01-01

    Helicobacter pylori vacuolating toxin (VacA) causes vacuolation in a variety of cultured cell lines, sensitivity to VacA differing greatly, however, among the different cell types. We found that the high sensitivity of HEp-2 cells to VacA was impaired by treating the cells with phosphatidylinositol-specific phospholipase C (PI-PLC) which removes glycosylphosphatidylinositol (GPI)-anchored proteins from the cell surface. Incubation of cells with a cholesterol-sequestering agent, that impairs both structure and function of sphingolipid-cholesterol-rich membrane microdomains (“lipid rafts”), also impaired VacA-induced cell vacuolation. Overexpression into HEp-2 cells of proteins inhibiting clathrin-dependent endocytosis (i.e., a dominant-negative mutant of Eps15, the five tandem Src-homology-3 domains of intersectin, and the K44A dominant-negative mutant of dynamin II) did not affect vacuolation induced by VacA. Nevertheless, F-actin depolymerization, known to block the different types of endocytic mechanisms, strongly impaired VacA vacuolating activity. Taken together, our data suggest that the high cell sensitivity to VacA depends on the presence of one or several GPI-anchored protein(s), intact membrane lipid rafts, and an uptake mechanism via a clathrin-independent endocytic pathway. PMID:11071915

  16. The calmodulin-like proteins AtCML4 and AtCML5 are single-pass membrane proteins targeted to the endomembrane system by an N-terminal signal anchor sequence.

    Science.gov (United States)

    Ruge, Henning; Flosdorff, Sandra; Ebersberger, Ingo; Chigri, Fatima; Vothknecht, Ute C

    2016-06-01

    Calmodulins (CaMs) are important mediators of Ca(2+) signals that are found ubiquitously in all eukaryotic organisms. Plants contain a unique family of calmodulin-like proteins (CMLs) that exhibit greater sequence variance compared to canonical CaMs. The Arabidopsis thaliana proteins AtCML4 and AtCML5 are members of CML subfamily VII and possess a CaM domain comprising the characteristic double pair of EF-hands, but they are distinguished from other members of this subfamily and from canonical CaMs by an N-terminal extension of their amino acid sequence. Transient expression of yellow fluorescent protein-tagged AtCML4 and AtCML5 under a 35S-promoter in Nicotiana benthamiana leaf cells revealed a spherical fluorescence pattern. This pattern was confirmed by transient expression in Arabidopsis protoplasts under the native promoter. Co-localization analyses with various endomembrane marker proteins suggest that AtCML4 and AtCML5 are localized to vesicular structures in the interphase between Golgi and the endosomal system. Further studies revealed AtCML5 to be a single-pass membrane protein that is targeted into the endomembrane system by an N-terminal signal anchor sequence. Self-assembly green fluorescent protein and protease protection assays support a topology with the CaM domain exposed to the cytosolic surface and not the lumen of the vesicles, indicating that AtCML5 could sense Ca(2+) signals in the cytosol. Phylogenetic analysis suggests that AtCML4 and AtCML5 are closely related paralogues originating from a duplication event within the Brassicaceae family. CML4/5-like proteins seem to be universally present in eudicots but are absent in some monocots. Together these results show that CML4/5-like proteins represent a flowering plant-specific subfamily of CMLs with a potential function in vesicle transport within the plant endomembrane system.

  17. 日本血吸虫糖基化磷脂酰肌醇锚定蛋白的鉴定%Identification of Glycosylphosphatidylinositol-anchored Protein from Schistosoma japonicum

    Institute of Scientific and Technical Information of China (English)

    曹勤燕; 薛艳凤; 沈利

    2012-01-01

    目的 鉴定日本血吸虫的糖基化磷脂酰肌醇锚定蛋白(GPI-AP). 方法 根据曼氏血吸虫的糖基化磷脂酰肌醇(GPI)锚定蛋白Sm200的编码基因(GenBank登录号为XM 002569560.1),运用生物信息学方法寻找日本血吸虫的同源基因,分析后选取基因蛋白质编码区(CDS)的部分基因序列(SjGPIs,长约933 bp)进行PCR扩增,并克隆入原核表达载体pET-28a(+),重组质粒转化大肠埃希菌(E.coli) BL21 (DE3)感受态细胞,异丙基-β-D-硫代半乳糖苷(IPTG)诱导表达,用镍柱Ni-NTA亲和层析纯化重组肽段SjGPIs.用纯化的重组肽段SjGPIs免疫新西兰大耳兔,以制备的重组肽段抗血清检测日本血吸虫GPI锚定蛋白.用磷脂酰肌醇特异性磷脂酶C(PI-PLC)鉴定检测到的蛋白在日本血吸虫虫体上的锚定方式.检测日本血吸虫感染小鼠的白细胞,确定其是否吞噬GPI锚定蛋白. 结果 日本血吸虫的基因组中存在与曼氏血吸虫GPI锚定蛋白Sm200基因的同源基因序列,经比对拼接后获得3495 bp含完整编码蛋白C末端的基因编码序列.以所选基因序列进行肽段原核表达,获得重组质粒pET-28a(+)-SjGPIs.通过对蛋白C末端序列分析、经蛋白质印迹(Western blotting)分析和PI-PLC酶切验证,发现日本血吸虫被膜存在以GPI形式锚定的蛋白,相对分子质量约为Mr 200000,命名为SjGPI200.感染日本血吸虫小鼠的白细胞中可检测到完整的SjGPI200蛋白. 结论 日本血吸虫存在锚定蛋白SjGPI200,并以GPI形式锚定于虫体被膜上.%Objective To identify glycosylphosphatidylinositol (GPI) anchored protein of Schistosoma japonicum. Methods Based on the gene sequence of Schistosoma mansoni GPI anchored protein Sm200 (GenBank Assess No: XM_002569560.1), bioinformatics analysis was performed to find out its homologous gene sequence in S. japonicum, then a selected partial coding sequence (SjGPIs, about 933 bp) from the homologous gene sequence were amplified, and

  18. Anchoring visions in organizations

    DEFF Research Database (Denmark)

    Simonsen, Jesper

    1999-01-01

    This paper introduces the term 'anchoring' within systems development: Visions, developed through early systems design within an organization, need to be deeply rooted in the organization. A vision's rationale needs to be understood by those who decide if the vision should be implemented as well...... as by those involved in the actual implementation. A model depicting a recent trend within systems development is presented: Organizations rely on purchasing generic software products and/or software development outsourced to external contractors. A contemporary method for participatory design, where...

  19. Anchoring a Leviathan: How the Nuclear Membrane Tethers the Genome.

    Science.gov (United States)

    Czapiewski, Rafal; Robson, Michael I; Schirmer, Eric C

    2016-01-01

    It is well established that the nuclear envelope has many distinct direct connections to chromatin that contribute to genome organization. The functional consequences of genome organization on gene regulation are less clear. Even less understood is how interactions of lamins and nuclear envelope transmembrane proteins (NETs) with chromatin can produce anchoring tethers that can withstand the physical forces of and on the genome. Chromosomes are the largest molecules in the cell, making megadalton protein structures like the nuclear pore complexes and ribosomes seem small by comparison. Thus to withstand strong forces from chromosome dynamics an anchoring tether is likely to be much more complex than a single protein-protein or protein-DNA interaction. Here we will briefly review known NE-genome interactions that likely contribute to spatial genome organization, postulate in the context of experimental data how these anchoring tethers contribute to gene regulation, and posit several hypotheses for the physical nature of these tethers that need to be investigated experimentally. Significantly, disruption of these anchoring tethers and the subsequent consequences for gene regulation could explain how mutations in nuclear envelope proteins cause diseases ranging from muscular dystrophy to lipodystrophy to premature aging progeroid syndromes. The two favored hypotheses for nuclear envelope protein involvement in disease are (1) weakening nuclear and cellular mechanical stability, and (2) disrupting genome organization and gene regulation. Considerable experimental support has been obtained for both. The integration of both mechanical and gene expression defects in the disruption of anchoring tethers could provide a unifying hypothesis consistent with both.

  20. Intelligent Educational Systems for Anchored Instruction?

    Science.gov (United States)

    Kumar, David D.

    1995-01-01

    Explores the potential for using Intelligent Educational Systems (IES) for anchoring instruction in macro contexts in science education. Topics include anchored instruction; situated cognition; problem solving; cognitivism; interactive video environments; and examples of combining IES and anchored instruction. (LRW)

  1. Release of Glycoprotein (GP1 from the Tegumental Surface of Taenia solium by Phospholipase C from Clostridium perfringens Suggests a Novel Protein-Anchor to Membranes

    Directory of Open Access Journals (Sweden)

    Abraham Landa

    2010-01-01

    Full Text Available In order to explore how molecules are linked to the membrane surface in larval Taenia solium, whole cysticerci were incubated in the presence of phospholipase C from Clostridium perfringens (PLC. Released material was collected and analyzed in polyacrylamide gels with sodium dodecyl sulfate. Two major bands with apparent molecular weights of 180 and 43 kDa were observed. Western blot of released material and localization assays in cysticerci tissue sections using antibodies against five known surface glycoproteins of T. solium cysticerci indicated that only one, previously called GP1, was released. Similar localization studies using the lectins wheat-germ-agglutinin and Concanavalin A showed that N-acetyl-D-glucosamine, N-acetylneuraminic, sialic acid, αmethyl-D-mannoside, D-manose/glucose, and N-acetyl-D-glucosamine residues are abundantly present on the surface. On the other hand, we find that treatment with PLC releases molecules from the surface; they do not reveal Cross Reacting Determinant (CRD, suggesting a novel anchor to the membrane for the glycoprotein GP1.

  2. Monitoring lipid anchor organization in cell membranes by PIE-FCCS.

    Science.gov (United States)

    Triffo, Sara B; Huang, Hector H; Smith, Adam W; Chou, Eldon T; Groves, Jay T

    2012-07-04

    This study examines the dynamic co-localization of lipid-anchored fluorescent proteins in living cells using pulsed-interleaved excitation fluorescence cross-correlation spectroscopy (PIE-FCCS) and fluorescence lifetime analysis. Specifically, we look at the pairwise co-localization of anchors from lymphocyte cell kinase (LCK: myristoyl, palmitoyl, palmitoyl), RhoA (geranylgeranyl), and K-Ras (farnesyl) proteins in different cell types. In Jurkat cells, a density-dependent increase in cross-correlation among RhoA anchors is observed, while LCK anchors exhibit a more moderate increase and broader distribution. No correlation was detected among K-Ras anchors or between any of the different anchor types studied. Fluorescence lifetime data reveal no significant Förster resonance energy transfer in any of the data. In COS 7 cells, minimal correlation was detected among LCK or RhoA anchors. Taken together, these observations suggest that some lipid anchors take part in anchor-specific co-clustering with other existing clusters of native proteins and lipids in the membrane. Importantly, these observations do not support a simple interpretation of lipid anchor-mediated organization driven by partitioning based on binary lipid phase separation.

  3. Binding constants of membrane-anchored receptors and ligands depend strongly on the nanoscale roughness of membranes.

    Science.gov (United States)

    Hu, Jinglei; Lipowsky, Reinhard; Weikl, Thomas R

    2013-09-17

    Cell adhesion and the adhesion of vesicles to the membranes of cells or organelles are pivotal for immune responses, tissue formation, and cell signaling. The adhesion processes depend sensitively on the binding constant of the membrane-anchored receptor and ligand proteins that mediate adhesion, but this constant is difficult to measure in experiments. We have investigated the binding of membrane-anchored receptor and ligand proteins with molecular dynamics simulations. We find that the binding constant of the anchored proteins strongly decreases with the membrane roughness caused by thermally excited membrane shape fluctuations on nanoscales. We present a theory that explains the roughness dependence of the binding constant for the anchored proteins from membrane confinement and that relates this constant to the binding constant of soluble proteins without membrane anchors. Because the binding constant of soluble proteins is readily accessible in experiments, our results provide a useful route to compute the binding constant of membrane-anchored receptor and ligand proteins.

  4. The membrane bound LRR lipoprotein Slr, and the cell wall-anchored M1 protein from Streptococcus pyogenes both interact with type I collagen.

    Directory of Open Access Journals (Sweden)

    Marta Bober

    Full Text Available Streptococcus pyogenes is an important human pathogen and surface structures allow it to adhere to, colonize and invade the human host. Proteins containing leucine rich repeats (LRR have been identified in mammals, viruses, archaea and several bacterial species. The LRRs are often involved in protein-protein interaction, are typically 20-30 amino acids long and the defining feature of the LRR motif is an 11-residue sequence LxxLxLxxNxL (x being any amino acid. The streptococcal leucine rich (Slr protein is a hypothetical lipoprotein that has been shown to be involved in virulence, but at present no ligands for Slr have been identified. We could establish that Slr is a membrane attached horseshoe shaped lipoprotein by homology modeling, signal peptidase II inhibition, electron microscopy (of bacteria and purified protein and immunoblotting. Based on our previous knowledge of LRR proteins we hypothesized that Slr could mediate binding to collagen. We could show by surface plasmon resonance that recombinant Slr and purified M1 protein bind with high affinity to collagen I. Isogenic slr mutant strain (MB1 and emm1 mutant strain (MC25 had reduced binding to collagen type I as shown by slot blot and surface plasmon resonance. Electron microscopy using gold labeled Slr showed multiple binding sites to collagen I, both to the monomeric and the fibrillar structure, and most binding occurred in the overlap region of the collagen I fibril. In conclusion, we show that Slr is an abundant membrane bound lipoprotein that is co-expressed on the surface with M1, and that both these proteins are involved in recruiting collagen type I to the bacterial surface. This underlines the importance of S. pyogenes interaction with extracellular matrix molecules, especially since both Slr and M1 have been shown to be virulence factors.

  5. Anchored Instruction and Situated Cognition Revisited.

    Science.gov (United States)

    Educational Technology, 1993

    1993-01-01

    Reviews theories of anchored instruction and addresses issues related to learning, transfer, and assessment. Highlights include video-based macrocontexts; videodisc anchors versus computer-based simulations; cooperative learning; transfer outside the classroom; authenticity; visual anchors versus verbal anchors; situated cognition; and using…

  6. Biogenesis of photosynthetic complexes in the chloroplast of Chlamydomonas reinhardtii requires ARSA1, a homolog of prokaryotic arsenite transporter and eukaryotic TRC40 for guided entry of tail-anchored proteins.

    Science.gov (United States)

    Formighieri, Cinzia; Cazzaniga, Stefano; Kuras, Richard; Bassi, Roberto

    2013-03-01

    as1, for antenna size mutant 1, was obtained by insertion mutagenesis of the unicellular green alga Chlamydomonas reinhardtii. This strain has a low chlorophyll content, 8% with respect to the wild type, and displays a general reduction in thylakoid polypeptides. The mutant was found to carry an insertion into a homologous gene, prokaryotic arsenite transporter (ARSA), whose yeast and mammal counterparts were found to be involved in the targeting of tail-anchored (TA) proteins to cytosol-exposed membranes, essential for several cellular functions. Here we present the characterization in a photosynthetic organism of an insertion mutant in an ARSA-homolog gene. The ARSA1 protein was found to be localized in the cytosol, and yet its absence in as1 leads to a small chloroplast and a strongly decreased chlorophyll content per cell. ARSA1 appears to be required for optimal biogenesis of photosynthetic complexes because of its involvement in the accumulation of TOC34, an essential component of the outer chloroplast membrane translocon (TOC) complex, which, in turn, catalyzes the import of nucleus-encoded precursor polypeptides into the chloroplast. Remarkably, the effect of the mutation appears to be restricted to biogenesis of chlorophyll-binding polypeptides and is not compensated by the other ARSA homolog encoded by the C. reinhardtii genome, implying a non-redundant function.

  7. Membrane curvature enables N-Ras lipid anchor sorting to liquid-ordered membrane phases

    DEFF Research Database (Denmark)

    Larsen, Jannik Bruun; Jensen, Martin Borch; Bhatia, Vikram Kjøller;

    2015-01-01

    Trafficking and sorting of membrane-anchored Ras GTPases are regulated by partitioning between distinct membrane domains. Here, in vitro experiments and microscopic molecular theory reveal membrane curvature as a new modulator of N-Ras lipid anchor and palmitoyl chain partitioning. Membrane...... curvature was essential for enrichment in raft-like liquid-ordered phases; enrichment was driven by relief of lateral pressure upon anchor insertion and most likely affects the localization of lipidated proteins in general....

  8. Microgravity Drill and Anchor System

    Science.gov (United States)

    Parness, Aaron; Frost, Matthew A.; King, Jonathan P.

    2013-01-01

    This work is a method to drill into a rock surface regardless of the gravitational field or orientation. The required weight-on-bit (WOB) is supplied by a self-contained anchoring mechanism. The system includes a rotary percussive coring drill, forming a complete sampling instrument usable by robot or human. This method of in situ sample acquisition using micro - spine anchoring technology enables several NASA mission concepts not currently possible with existing technology, including sampling from consolidated rock on asteroids, providing a bolt network for astronauts visiting a near-Earth asteroid, and sampling from the ceilings or vertical walls of lava tubes and cliff faces on Mars. One of the most fundamental parameters of drilling is the WOB; essentially, the load applied to the bit that allows it to cut, creating a reaction force normal to the surface. In every drilling application, there is a minimum WOB that must be maintained for the system to function properly. In microgravity (asteroids and comets), even a small WOB could not be supported conventionally by the weight of the robot or astronaut. An anchoring mechanism would be needed to resist the reactions, or the robot or astronaut would push themselves off the surface and into space. The ability of the system to anchor itself to a surface creates potential applications that reach beyond use in low gravity. The use of these anchoring mechanisms as end effectors on climbing robots has the potential of vastly expanding the scope of what is considered accessible terrain. Further, because the drill is supported by its own anchor rather than by a robotic arm, the workspace is not constrained by the reach of such an arm. Yet, if the drill is on a robotic arm, it has the benefit of not reflecting the forces of drilling back to the arm s joints. Combining the drill with the anchoring feet will create a highly mobile, highly stable, and highly reliable system. The drilling system s anchor uses hundreds of

  9. N-terminus of IpaB provides a potential anchor to the Shigella type III secretion system tip complex protein IpaD.

    Science.gov (United States)

    Dickenson, Nicholas E; Arizmendi, Olivia; Patil, Mrinalini K; Toth, Ronald T; Middaugh, C Russell; Picking, William D; Picking, Wendy L

    2013-12-10

    The type III secretion system (T3SS) is an essential virulence factor for Shigella flexneri , providing a conduit through which host-altering effectors are injected directly into a host cell to promote uptake. The type III secretion apparatus (T3SA) is composed of a basal body, external needle, and regulatory tip complex. The nascent needle is a polymer of MxiH capped by a pentamer of invasion plasmid antigen D (IpaD). Exposure to bile salts (e.g., deoxycholate) causes a conformational change in IpaD and promotes recruitment of IpaB to the needle tip. It has been proposed that IpaB senses contact with host cell membranes, recruiting IpaC and inducing full secretion of T3SS effectors. Although the steps of T3SA maturation and their external triggers have been identified, details of specific protein interactions and mechanisms have remained difficult to study because of the hydrophobic nature of the IpaB and IpaC translocator proteins. Here, we explored the ability for a series of soluble N-terminal IpaB peptides to interact with IpaD. We found that DOC is required for the interaction and that a region of IpaB between residues 11-27 is required for maximum binding, which was confirmed in vivo. Furthermore, intramolecular FRET measurements indicated that movement of the IpaD distal domain away from the protein core accompanied the binding of IpaB11-226. Together, these new findings provide important new insight into the interactions and potential mechanisms that define the maturation of the Shigella T3SA needle tip complex and provide a foundation for further studies probing T3SS activation.

  10. Construction of a novel cell-surface display system for heterologous gene expression in Escherichia coli by using an outer membrane protein of Zymomonas mobilis as anchor motif.

    Science.gov (United States)

    He, Ming-Xiong; Feng, Hong; Zhang, Yi-Zheng

    2008-12-01

    A novel bacterial cell-surface display system was developed in Escherichia coli using omp1, a hypothetical outer membrane protein of Zymomonas mobilis. By using this system, we successfully expressed beta-amylase gene of sweet potato in E. coli. The display of enzyme on the membrane surface was also confirmed. The recombinant beta-amylase showed to significantly increase hydrolytic activity toward soluble starch. Our results provide a basis for constructing an engineered Z. mobilis strain directly fermenting raw starch to produce ethanol.

  11. Anchoring a Leviathan: How the Nuclear Membrane Tethers the Genome

    Directory of Open Access Journals (Sweden)

    Rafal eCzapiewski

    2016-05-01

    Full Text Available It is well established that the nuclear envelope has many distinct direct connections to chromatin that contribute to genome organization. The functional consequences of genome organization on gene regulation are less clear. Even less understood is how interactions of lamins and nuclear envelope transmembrane proteins (NETs with chromatin can produce anchoring tethers that can withstand the physical forces of and on the genome. Chromosomes are the largest molecules in the cell, making megadalton protein structures like the nuclear pore complexes and ribosomes seem small by comparison. Thus to withstand strong forces from chromosome dynamics an anchoring tether is likely to be much more complex than a single protein-protein or protein-DNA interaction. Here we will briefly review known NE-genome interactions that likely contribute to spatial genome organization, postulate in the context of experimental data how these anchoring tethers contribute to gene regulation, and posit several hypotheses for the physical nature of these tethers that need to be investigated experimentally. Significantly, disruption of these anchoring tethers and the subsequent consequences for gene regulation could explain how mutations in nuclear envelope proteins cause diseases ranging from muscular dystrophy to lipodystrophy to premature ageing progeroid syndromes. The two favored hypotheses for nuclear envelope protein involvement in disease are 1 weakening nuclear and cellular mechanical stability, and 2 disrupting genome organization and gene regulation. Considerable experimental support has been obtained for both. The integration of both mechanical and gene expression defects in the disruption of anchoring tethers could provide a unifying hypothesis consistent with both.

  12. SwissProt search result: AK240924 [KOME

    Lifescience Database Archive (English)

    Full Text Available AK240924 J065039N21 (Q99996) A-kinase anchor protein 9 (Protein kinase A anchoring ...protein 9) (PRKA9) (A-kinase anchor protein 450 kDa) (AKAP 450) (A-kinase anchor protein 350 kDa) (AKAP 350)

  13. Insect cell-derived cofactors become fully functional after proteinase K and heat treatment for high-fidelity amplification of glycosylphosphatidylinositol-anchored recombinant scrapie and BSE prion proteins.

    Science.gov (United States)

    Imamura, Morikazu; Kato, Nobuko; Okada, Hiroyuki; Yoshioka, Miyako; Iwamaru, Yoshifumi; Shimizu, Yoshihisa; Mohri, Shirou; Yokoyama, Takashi; Murayama, Yuichi

    2013-01-01

    The central event in prion infection is the conformational conversion of host-encoded cellular prion protein (PrP(C)) into the pathogenic isoform (PrP(Sc)). Diverse mammalian species possess the cofactors required for in vitro replication of PrP(Sc) by protein-misfolding cyclic amplification (PMCA), but lower organisms, such as bacteria, yeasts, and insects, reportedly lack the essential cofactors. Various cellular components, such as RNA, lipids, and other identified cofactor molecules, are commonly distributed in both eukaryotes and prokaryotes, but the reasons for the absence of cofactor activity in lower organisms remain to be elucidated. Previously, we reported that brain-derived factors were necessary for the in vitro replication of glycosylphosphatidylinositol-anchored baculovirus-derived recombinant PrP (Bac-PrP). Here, we demonstrate that following protease digestion and heat treatment, insect cell lysates had the functional cofactor activity required for Bac-PrP replication by PMCA. Mammalian PrP(Sc) seeds and Bac-PrP(Sc) generated by PMCA using Bac-PrP and insect cell-derived cofactors showed similar pathogenicity and produced very similar lesions in the brains of inoculated mice. These results suggested that the essential cofactors required for the high-fidelity replication of mammalian PrP(Sc) were present in the insect cells but that the cofactor activity was masked or inhibited in the native state. We suggest that not only RNA, but also DNA, are the key components of PMCA, although other cellular factors were necessary for the expression of the cofactor activity of nucleic acids. PMCA using only insect cell-derived substances (iPMCA) was highly useful for the ultrasensitive detection of PrP(Sc) of some prion strains.

  14. Seals, Concrete Anchors, and Connections

    Science.gov (United States)

    1989-02-01

    the system the cable sizes can be changed even after concreting work _ is finished. i W The structure is also suitable for modern concrete formwork ...1 ruiinn 0i all 3up-H-Stud is a heavy-duty, all steel, expansion wedge anchor types of equipment. Typical applications: tunnel liner panels, air

  15. Expression of GPI-80, a beta2-integrin-associated glycosylphosphatidylinositol-anchored protein, requires neutrophil differentiation with dimethyl sulfoxide in HL-60 cells.

    Science.gov (United States)

    Takeda, Yuji; Fu, Junfen; Suzuki, Kichiya; Sendo, Dai; Nitto, Takeaki; Sendo, Fujiro; Araki, Yoshihiko

    2003-06-10

    GPI-80 is a member of the amidohydrolase family that has been proposed as a potential regulator of beta2-integrin-dependent leukocyte adhesion. GPI-80 is expressed mainly in human neutrophils. Our previous studies suggested that GPI-80 expression might be associated with myeloid differentiation. To verify this, we examined whether GPI-80 is expressed on the human promyelocytic leukemia cell line HL-60 following treatment with differentiation inducers. GPI-80 expression was induced in cells treated with dimethyl sulfoxide (DMSO) to stimulate differentiation down the neutrophil pathway. On the other hand, all-trans-retinoic acid (ATRA), another neutrophil-inducing reagent, induced no clear GPI-80 expression. Potent monocyte-inducing reagents such as 1alpha,25-dihydroxyvitamin D(3) or phorbol 12-myristate 13-acetate also had no significant effect on the protein expression. GPI-80-positive cells were found in the well-differentiated CD11b-positive and transferrin-receptor-negative cell population. Granulocyte colony-stimulating factor, which augments neutrophil differentiation of HL-60 cells, up-regulated GPI-80 expression in the presence of DMSO. Granulocyte/macrophage colony-stimulating factor, which is known to suppress the neutrophil maturation of cells, inhibited expression. Adhesion of DMSO-induced cells was regulated by anti-GPI-80 monoclonal antibody, similar to the regulation observed in neutrophils. These results suggest that use of DMSO to induce neutrophil differentiation provides suitable conditions for GPI-80 expression, and that this culture system may be a helpful model for further study of the regulation of GPI-80 expression during myeloid differentiation.

  16. Structural models of the membrane anchors of envelope glycoproteins E1 and E2 from pestiviruses

    Energy Technology Data Exchange (ETDEWEB)

    Wang, Jimin, E-mail: jimin.wang@yale.edu; Li, Yue; Modis, Yorgo, E-mail: yorgo.modis@yale.edu

    2014-04-15

    The membrane anchors of viral envelope proteins play essential roles in cell entry. Recent crystal structures of the ectodomain of envelope protein E2 from a pestivirus suggest that E2 belongs to a novel structural class of membrane fusion machinery. Based on geometric constraints from the E2 structures, we generated atomic models of the E1 and E2 membrane anchors using computational approaches. The E1 anchor contains two amphipathic perimembrane helices and one transmembrane helix; the E2 anchor contains a short helical hairpin stabilized in the membrane by an arginine residue, similar to flaviviruses. A pair of histidine residues in the E2 ectodomain may participate in pH sensing. The proposed atomic models point to Cys987 in E2 as the site of disulfide bond linkage with E1 to form E1–E2 heterodimers. The membrane anchor models provide structural constraints for the disulfide bonding pattern and overall backbone conformation of the E1 ectodomain. - Highlights: • Structures of pestivirus E2 proteins impose constraints on E1, E2 membrane anchors. • Atomic models of the E1 and E2 membrane anchors were generated in silico. • A “snorkeling” arginine completes the short helical hairpin in the E2 membrane anchor. • Roles in pH sensing and E1–E2 disulfide bond formation are proposed for E1 residues. • Implications for E1 ectodomain structure and disulfide bonding pattern are discussed.

  17. 24 CFR 3285.401 - Anchoring instructions.

    Science.gov (United States)

    2010-04-01

    ... wind by use of anchor assembly type installations or by connecting the home to an alternative foundation system. See § 3285.301. (b) For anchor assembly type installations, the installation instructions... instructions and design for anchor type assemblies must be prepared by a registered professional engineer...

  18. Rigid rod anchored to infinite membrane.

    Science.gov (United States)

    Guo, Kunkun; Qiu, Feng; Zhang, Hongdong; Yang, Yuliang

    2005-08-15

    We investigate the shape deformation of an infinite membrane anchored by a rigid rod. The density profile of the rod is calculated by the self-consistent-field theory and the shape of the membrane is predicted by the Helfrich membrane elasticity theory [W. Helfrich, Z. Naturforsch. 28c, 693 (1973)]. It is found that the membrane bends away from the rigid rod when the interaction between the rod and the membrane is repulsive or weakly attractive (adsorption). However, the pulled height of the membrane at first increases and then decreases with the increase of the adsorption strength. Compared to a Gaussian chain with the same length, the rigid rod covers much larger area of the membrane, whereas exerts less local entropic pressure on the membrane. An evident gap is found between the membrane and the rigid rod because the membrane's curvature has to be continuous. These behaviors are compared with that of the flexible-polymer-anchored membranes studied by previous Monte Carlo simulations and theoretical analysis. It is straightforward to extend this method to more complicated and real biological systems, such as infinite membrane/multiple chains, protein inclusion, or systems with phase separation.

  19. Anchoring Bias in Online Voting

    CERN Document Server

    Yang, Zimo; Zhou, Tao

    2012-01-01

    Voting online with explicit ratings could largely reflect people's preferences and objects' qualities, but ratings are always irrational, because they may be affected by many unpredictable factors like mood, weather, as well as other people's votes. By analyzing two real systems, this paper reveals a systematic bias embedding in the individual decision-making processes, namely people tend to give a low rating after a low rating, as well as a high rating following a high rating. This so-called \\emph{anchoring bias} is validated via extensive comparisons with null models, and numerically speaking, the extent of bias decays with interval voting number in a logarithmic form. Our findings could be applied in the design of recommender systems and considered as important complementary materials to previous knowledge about anchoring effects on financial trades, performance judgements, auctions, and so on.

  20. Inconspicuous anchoring effects generated by false information

    Institute of Scientific and Technical Information of China (English)

    Chen Qu; Jun Wang; Yuejia Luo

    2008-01-01

    The impact of false information on numerical judgments was examined on young normal subjects by an event-related potential (ERP) experiment. To imitate the judgments in real world, we ensured the subjects acknowledged of the target task. The behavioral results found that both uncertain information and false information assimilated the final estimates: higher after higher anchors and lower after lower anchors; and false information caused a weaker anchoring bias than uncertain information. ERP results provided further electrophysiological evidence for the mechanism of anchoring. In the early phrase, it was an accessibility-dominated process in which two kinds of anchors elicited an N300 component related to the accessibility of anchors propositions. The knowledge relevant to targets joined the process in the late phrase, which caused a larger amplitude of late positive component (LPC) for implausible lower anchors than that for plausible higher anchors. Source analysis showed that medial frontal gyrus, whose activity was suggested to signal the need of adjustment, was more reliable to explain the LPC elicited by implausible lower anchors. Therefore, we suggest that accessibility is facilitated when the external anchor is consistent with the world knowledge, and adjustment is initiated when the external anchor is inconsistent.

  1. Anchoring in Numeric Judgments of Visual Stimuli.

    Science.gov (United States)

    Langeborg, Linda; Eriksson, Mårten

    2016-01-01

    This article investigates effects of anchoring in age estimation and estimation of quantities, two tasks which to different extents are based on visual stimuli. The results are compared to anchoring in answers to classic general knowledge questions that rely on semantic knowledge. Cognitive load was manipulated to explore possible differences between domains. Effects of source credibility, manipulated by differing instructions regarding the selection of anchor values (no information regarding anchor selection, information that the anchors are randomly generated or information that the anchors are answers from an expert) on anchoring were also investigated. Effects of anchoring were large for all types of judgments but were not affected by cognitive load or by source credibility in either one of the researched domains. A main effect of cognitive load on quantity estimations and main effects of source credibility in the two visually based domains indicate that the manipulations were efficient. Implications for theoretical explanations of anchoring are discussed. In particular, because anchoring did not interact with cognitive load, the results imply that the process behind anchoring in visual tasks is predominantly automatic and unconscious.

  2. Upper section for anchor timbering

    Energy Technology Data Exchange (ETDEWEB)

    Shirokov, A.P.; Isachenko, V.M.; Kuntsevich, V.I.; Pishulin, V.V.

    1983-01-01

    The purpose of the invention is to exclude rotation of the fixing device around the anchor with screwing of the tightening nut and simplification of the fixing device design. The support fixing device is made of a section of pipe and is equipped with a rotation delimiter made in the form of a female fixing device of the projection arranged on it at an angle to the longitudinal axis.

  3. Simultaneous blockade of NFkappaB, JNK, and p38 MAPK by a kinase-inactive mutant of the protein kinase TAK1 sensitizes cells to apoptosis and affects a distinct spectrum of tumor necrosis factor [corrected] target genes.

    Science.gov (United States)

    Thiefes, Axel; Wolter, Sabine; Mushinski, J Frederic; Hoffmann, Elke; Dittrich-Breiholz, Oliver; Graue, Nadine; Dörrie, Anneke; Schneider, Heike; Wirth, Dagmar; Luckow, Bruno; Resch, Klaus; Kracht, Michael

    2005-07-29

    The inflammatory response is characterized by the induction (or repression) of hundreds of genes. The activity of many of these genes is controlled by MAPKs and the IkappaB kinase-NFkappaB pathway. To reveal the effects of blocking these pathways simultaneously, fibroblasts were infected with retroviruses encoding TAK1K63W, an inactive mutant of the protein kinase TAK1. Expression of this protein inhibited tumor necrosis factor (TNF)-induced activation of NFkappaB, JNK, and p38 MAPK and sensitized the cells to TNF-induced apoptosis. 23 different microarray experiments were used to analyze the expression of >7000 genes in these cells. We identified 518 genes that were regulated by TNF in both TAK1K63W-expressing cells and control cells, 37 genes induced by TNF only when TAK1K63W was present, and 48 TNF-induced genes that were suppressed by TAK1K63W. The TNF-inducible genes that were most strongly suppressed by TAK1K63W, ccl2, ccl7, ccl5, cxcl1, cxcl5, cxcl10, saa3, and slpi also had much lower basal levels of expression, indicating that TAK1 also played a role in their normal expression. Chromatin immunoprecipitation studies on four of these genes suggested that inactivation of TAK1 activity led to direct suppression of expression at the transcriptional level because of impaired recruitment of RNA polymerase II to their promoters. ccl2 induction by TNF or interleukin-1 was also suppressed in cells that expressed TAK1 antisense RNA or that were genetically deficient in JNK1/2 or p65 NFkappaB. These data suggest that regulation of the expression of a selected group of inflammation-related genes is funneled through TAK1, making it a potentially useful target for more specific anti-inflammatory drug development.

  4. A lunar/Martian anchor emplacement system

    Science.gov (United States)

    Clinton, Dustin; Holt, Andrew; Jantz, Erik; Kaufman, Teresa; Martin, James; Weber, Reed

    1993-01-01

    On the Moon or Mars, it is necessary to have an anchor, or a stable, fixed point able to support the forces necessary to rescue a stuck vehicle, act as a stake for a tent in a Martian gale, act as a fulcrum in the erection of general construction poles, or support tent-like regolith shields. The anchor emplacement system must be highly autonomous. It must supply the energy and stability for anchor deployment. The goal of the anchor emplacement system project is to design and build a prototype anchor and to design a conceptual anchor emplacement system. Various anchors were tested in a 1.3 cubic meter test bed containing decomposed granite. A simulated lunar soil was created by adjusting the moisture and compaction characteristics of the soil. We conducted tests on emplacement torque, amount of force the anchor could withstand before failure, anchor pull out force at various angles, and soil disturbances caused by placing the anchor. A single helix auger anchor performed best in this test bed based on energy to emplace, and the ultimate holding capacity. The anchor was optimized for ultimate holding capacity, minimum emplacement torque, and minimum soil disturbance in sandy soils yielding the following dimensions: helix diameter (4.45 cm), pitch (1.27 cm), blade thickness (0.15 cm), total length (35.56 cm), shaft diameter (0.78 cm), and a weight of 212.62 g. The experimental results showed that smaller diameter, single-helix augers held more force than larger diameter augers for a given depth. The emplacement system consists of a flywheel and a motor for power, sealed in a protective box supported by four legs. The flywheel system was chosen over a gear system based on its increased reliability in the lunar environment.

  5. Anchored Lagrangian submanifolds and their Floer theory

    CERN Document Server

    Fukaya, Kenji; Ohta, Hiroshi; Ono, Kaoru

    2009-01-01

    We introduce the notion of (graded) anchored Lagrangian submanifolds and use it to study the filtration of Floer' s chain complex. We then obtain an anchored version of Lagrangian Floer homology and its (higher) product structures. They are somewhat different from the more standard non-anchored version. The anchored version discussed in this paper is more naturally related to the variational picture of Lagrangian Floer theory and so to the likes of spectral invariants. We also discuss rationality of Lagrangian submanifold and reduction of the coefficient ring of Lagrangian Floer cohomology of thereof.

  6. Anchored Instruction and Anchored Assessment: An Ecological Approach to Measuring Situated Learning.

    Science.gov (United States)

    Young, Michael F.; Kulikowich, Jonna M.

    Anchored instruction and anchored assessment are described and illustrated through a mathematics problem from the Jasper problem solving series developed at Vanderbilt University in Nashville (Tennessee). Anchored instruction is instruction situated in a context complex enough to provide meaning and reasons for why information is useful. Problems…

  7. Anchor Bolt Position in Base Plate In Terms Of T and J Anchor Bolt

    Directory of Open Access Journals (Sweden)

    b Osman Mohamad Hairi

    2017-01-01

    Full Text Available Generally, L anchor bolt system has been used for a long period of time in construction industry as one of the distributing load structures. However, there are some weaknesses in L anchor bolt which may straighten and pullup when charged with tensile load. Current practices prefer to use other types of anchor bolt systems, such as headed studs anchor bolt system to replace the L anchor bolt design. There has been lack of studies to prove that it is more effective in terms of performance. A new T anchor bolt which was basically modified from headed studs anchor bolt was proposed in this study to compare its performance of tensile loading in concrete failure to typical L design. This study aims to determine whether the T anchor bolt system gives better performance as compared to an L anchor bolt system. The performance was rated based on tensile loading on concrete failure pattern. A pullout test was conducted on two different anchor bolt systems, namely L and T. The anchor bolt embedded depth, h in concrete were varied according to their hook or bend radius. Each sample was repeated twice. There were totally eight samples. The hook or bend radius used were 50 mm and 57.5 mm for sample L1 and L2, respectively. 90-degree bend were used on sample T1 and T2. Based on test results, it can be seen that the performance of concrete failure pattern under tensile load on both L and T anchor bolt design samples with 200 mm embedment depth was better than deeper embedment depth of 230 mm. But the L anchor bolt design gives the best results as compared to T design. Although T anchor bolt design shows higher resistance before first bond failure to the concrete sample. T anchor bolt was analysed and needed deeper embedment depth to allow formation of cone pull-out shape to acquire better performance.

  8. Anchored Instruction in a Situated Learning Environment.

    Science.gov (United States)

    Lee, Miwha

    The purpose of this study was to design and develop a multimedia-based anchored program and to examine the effects of students' and group characteristics on the problem-solving process in anchored instruction with the multimedia program in a situated learning environment. Sixty-eight students were assigned to small groups via a stratified random…

  9. DSSC anchoring groups: a surface dependent decision.

    Science.gov (United States)

    O'Rourke, C; Bowler, D R

    2014-05-14

    Electrodes in dye sensitised solar cells are typically nanocrystalline anatase TiO2 with a majority (1 0 1) surface exposed. Generally the sensitising dye employs a carboxylic anchoring moiety through which it adheres to the TiO₂ surface. Recent interest in exploiting the properties of differing TiO₂ electrode morphologies, such as rutile nanorods exposing the (1 1 0) surface and anatase electrodes with high percentages of the (0 0 1) surface exposed, begs the question of whether this anchoring strategy is best, irrespective of the majority surface exposed. Here we address this question by presenting density functional theory calculations contrasting the binding properties of two promising anchoring groups, phosphonic acid and boronic acid, to that of carboxylic acid. Anchor-electrode interactions are studied for the prototypical anatase (1 0 1) surface, along with the anatase (0 0 1) and rutile (1 1 0) surfaces. Finally the effect of using these alternative anchoring groups to bind a typical coumarin dye (NKX-2311) to these TiO₂ substrates is examined. Significant differences in the binding properties are found depending on both the anchor and surface, illustrating that the choice of anchor is necessarily dependent upon the surface exposed in the electrode. In particular the boronic acid is found to show the potential to be an excellent anchor choice for electrodes exposing the anatase (0 0 1) surface.

  10. Method of fabrication of anchored nanostructure materials

    Science.gov (United States)

    Seals, Roland D; Menchhofer, Paul A; Howe, Jane Y; Wang, Wei

    2013-11-26

    Methods for fabricating anchored nanostructure materials are described. The methods include heating a nano-catalyst under a protective atmosphere to a temperature ranging from about 450.degree. C. to about 1500.degree. C. and contacting the heated nano-catalysts with an organic vapor to affix carbon nanostructures to the nano-catalysts and form the anchored nanostructure material.

  11. The effect of accuracy motivation on anchoring and adjustment: do people adjust from provided anchors?

    Science.gov (United States)

    Simmons, Joseph P; LeBoeuf, Robyn A; Nelson, Leif D

    2010-12-01

    Increasing accuracy motivation (e.g., by providing monetary incentives for accuracy) often fails to increase adjustment away from provided anchors, a result that has led researchers to conclude that people do not effortfully adjust away from such anchors. We challenge this conclusion. First, we show that people are typically uncertain about which way to adjust from provided anchors and that this uncertainty often causes people to believe that they have initially adjusted too far away from such anchors (Studies 1a and 1b). Then, we show that although accuracy motivation fails to increase the gap between anchors and final estimates when people are uncertain about the direction of adjustment, accuracy motivation does increase anchor-estimate gaps when people are certain about the direction of adjustment, and that this is true regardless of whether the anchors are provided or self-generated (Studies 2, 3a, 3b, and 5). These results suggest that people do effortfully adjust away from provided anchors but that uncertainty about the direction of adjustment makes that adjustment harder to detect than previously assumed. This conclusion has important theoretical implications, suggesting that currently emphasized distinctions between anchor types (self-generated vs. provided) are not fundamental and that ostensibly competing theories of anchoring (selective accessibility and anchoring-and-adjustment) are complementary.

  12. Ringstone anchors from Gujarat, west coast of India

    Digital Repository Service at National Institute of Oceanography (India)

    Gaur, A.S.; Sundaresh; Tripati, S.; Bandodkar, S.N.

    of Dwarka and Somanath have yielded several ringstone anchors along with other stone anchors such as triangular and grapnel types. The raw material used for these ring stones comprises basalt, sandstone and limestone. Earlier, these anchors were identified...

  13. A Tenebrio molitor GPI-anchored alkaline phosphatase is involved in binding of Bacillus thuringiensis Cry3Aa to brush border membrane vesicles.

    Science.gov (United States)

    Zúñiga-Navarrete, Fernando; Gómez, Isabel; Peña, Guadalupe; Bravo, Alejandra; Soberón, Mario

    2013-03-01

    Bacillus thuringiensis Cry toxins recognizes their target cells in part by the binding to glycosyl-phosphatidyl-inositol (GPI) anchored proteins such as aminopeptidase-N (APN) or alkaline phosphatases (ALP). Treatment of Tenebrio molitor brush border membrane vesicles (BBMV) with phospholipase C that cleaves out GPI-anchored proteins from the membranes, showed that GPI-anchored proteins are involved in binding of Cry3Aa toxin to BBMV. A 68 kDa GPI-anchored ALP was shown to bind Cry3Aa by toxin overlay assays. The 68 kDa GPI-anchored ALP was preferentially expressed in early instar larvae in comparison to late instar larvae. Our work shows for the first time that GPI-anchored ALP is important for Cry3Aa binding to T. molitor BBMV suggesting that the mode of action of Cry toxins is conserved in different insect orders.

  14. Spatial regulation of the cAMP-dependent protein kinase during chemotactic cell migration.

    Science.gov (United States)

    Howe, Alan K; Baldor, Linda C; Hogan, Brian P

    2005-10-04

    Historically, the cAMP-dependent protein kinase (PKA) has a paradoxical role in cell motility, having been shown to both facilitate and inhibit actin cytoskeletal dynamics and cell migration. In an effort to understand this dichotomy, we show here that PKA is regulated in subcellular space during cell migration. Immunofluorescence microscopy and biochemical enrichment of pseudopodia showed that type II regulatory subunits of PKA and PKA activity are enriched in protrusive cellular structures formed during chemotaxis. This enrichment correlates with increased phosphorylation of key cytoskeletal substrates for PKA, including the vasodilator-stimulated phosphoprotein (VASP) and the protein tyrosine phosphatase containing a PEST motif. Importantly, inhibition of PKA activity or its ability to interact with A kinase anchoring proteins inhibited the activity of the Rac GTPase within pseudopodia. This effect correlated with both decreased guanine nucleotide exchange factor activity and increased GTPase activating protein activity. Finally, inhibition of PKA anchoring, like inhibition of total PKA activity, inhibited pseudopod formation and chemotactic cell migration. These data demonstrate that spatial regulation of PKA via anchoring is an important facet of normal chemotactic cell movement.

  15. Azimuthal anchoring of a nematic liquid crystal on a grooved interface with anisotropic polar anchoring

    Institute of Scientific and Technical Information of China (English)

    Zhou Xuan; Zhang Zhi-Dong; Ye Wen-Jiang; Xuan Li

    2012-01-01

    Zhang Y Jet al.[Zhang Y J,Zhang Z D,Zhu L Z and Xuan L 2011 Liquid Cryst.38 355] investigated the effects of finite polar anchoring on the azimuthal anchoring energy at a grooved interface,in which polar anchoring was isotropic in the local tangent plane of the surface.In this paper,we investigate the effects of both isotropic and anisotropic polar anchoring on the surface anchoring energy in the frame of Fukuda et al.'s theory.The results show that anisotropic polar anchoring strengthens the azimuthal anchoring of grooved surfaces.In the one-elastic-constant approximation (K11 =K22 =K33 =K),the surface-groove-induced azimuthal anchoring energy is entirely consistent with the result of Faetti,and it reduces to the original result of Berreman with an increase in polar anchoring.Moreover,the contribution of the surface-like elastic term to the Rapini-Papoular anchoring energy is zero.

  16. Anchored nanostructure materials and method of fabrication

    Science.gov (United States)

    Seals, Roland D; Menchhofer, Paul A; Howe, Jane Y; Wang, Wei

    2012-11-27

    Anchored nanostructure materials and methods for their fabrication are described. The anchored nanostructure materials may utilize nano-catalysts that include powder-based or solid-based support materials. The support material may comprise metal, such as NiAl, ceramic, a cermet, or silicon or other metalloid. Typically, nanoparticles are disposed adjacent a surface of the support material. Nanostructures may be formed as anchored to nanoparticles that are adjacent the surface of the support material by heating the nano-catalysts and then exposing the nano-catalysts to an organic vapor. The nanostructures are typically single wall or multi-wall carbon nanotubes.

  17. Identification of Ski as a target for Aurora A kinase.

    Science.gov (United States)

    Mosquera, Jocelyn; Armisen, Ricardo; Zhao, Hongling; Rojas, Diego A; Maldonado, Edio; Tapia, Julio C; Colombo, Alicia; Hayman, Michael J; Marcelain, Katherine

    2011-06-10

    Ski is a negative regulator of the transforming growth factor-β and other signalling pathways. The absence of SKI in mouse fibroblasts leads to chromosome segregation defects and genomic instability, suggesting a role for Ski during mitosis. At this stage, Ski is phosphorylated but to date little is known about the kinases involved in this process. Here, we show that Aurora A kinase is able to phosphorylate Ski in vitro. In vivo, Aurora A and Ski co-localized at the centrosomes and co-immunoprecipitated. Conversely, a C-terminal truncation mutant of Ski (SkiΔ491-728) lacking a coiled-coil domain, displayed decreased centrosomal localization. This mutant no longer co-immunoprecipitated with Aurora-A in vivo, but was still phosphorylated in vitro, indicating that the Ski-Aurora A interaction takes place at the centrosomes. These data identify Ski as a novel target of Aurora A and contribute to an understanding of the role of these proteins in the mitotic process.

  18. Starting point anchoring effects in choice experiments

    DEFF Research Database (Denmark)

    Ladenburg, Jacob; Olsen, Søren Bøye

    of preferences in Choice Experiments resembles the Dichotomous Choice format, there is reason to suspect that Choice Experiments are equally vulnerable to anchoring bias. Employing different sets of price levels in a so-called Instruction Choice Set presented prior to the actual choice sets, the present study......Anchoring is acknowledged as a potential source of considerable bias in Dichotomous Choice Contingent Valuation studies. Recently, another stated preference method known as Choice Experiments has gained in popularity as well as the number of applied studies. However, as the elicitation...... finds that preferences elicited by Choice Experiments can be subject to starting point anchoring bias. Different price levels provoked significantly different distributions of choice in two otherwise identical choice set designs. On a more specific level, the results indicate that the anchoring...

  19. Starting point anchoring effects in choice experiments

    DEFF Research Database (Denmark)

    Ladenburg, Jacob; Olsen, Søren Bøye

    Anchoring is acknowledged as a potential source of considerable bias in Dichotomous Choice Contingent Valuation studies. Recently, another stated preference method known as Choice Experiments has gained in popularity as well as the number of applied studies. However, as the elicitation...... of preferences in Choice Experiments resembles the Dichotomous Choice format, there is reason to suspect that Choice Experiments are equally vulnerable to anchoring bias. Employing different sets of price levels in a so-called Instruction Choice Set presented prior to the actual choice sets, the present study...... finds that preferences elicited by Choice Experiments can be subject to starting point anchoring bias. Different price levels provoked significantly different distributions of choice in two otherwise identical choice set designs. On a more specific level, the results indicate that the anchoring...

  20. The Use of Comics-Based Cases in Anchored Instruction

    Science.gov (United States)

    Kneller, Matthew F.

    2009-01-01

    The primary purpose of this research was to understand how comics fulfill the role of anchor in an anchored instruction learning environment. Anchored instruction addresses the inert knowledge problem through the use of realistic multimedia stories, or "anchors," that embed a problem and the necessary data to solve it within the narrative. In the…

  1. Anchors as Semantic Primes in Value Construction: An EEG Study of the Anchoring Effect.

    Science.gov (United States)

    Ma, Qingguo; Li, Diandian; Shen, Qiang; Qiu, Wenwei

    2015-01-01

    Previous research regarding anchoring effects has demonstrated that human judgments are often assimilated to irrelevant information. Studies have demonstrated that anchors influence the economic valuation of various products and experiences; however, the cognitive explanations of this effect remain controversial, and its neural mechanisms have rarely been explored. In the current study, we conducted an electroencephalography (EEG) experiment to investigate the anchoring effect on willingness to accept (WTA) for an aversive hedonic experience and the role of anchors in this judgment heuristic. The behavioral results demonstrated that random numbers affect participants' WTA for listening to pieces of noise. The participants asked for higher pay after comparing their WTA with higher numbers. The EEG results indicated that anchors also influenced the neural underpinnings of the valuation process. Specifically, when a higher anchor number was drawn, larger P2 and late positive potential amplitudes were elicited, reflecting the anticipation of more intensive pain from the subsequent noise. Moreover, higher anchors induced a stronger theta band power increase compared with lower anchors when subjects listened to the noises, indicating that the participants felt more unpleasant during the actual experience of the noise. The levels of unpleasantness during both anticipation and experience were consistent with the semantic information implied by the anchors. Therefore, these data suggest that a semantic priming process underlies the anchoring effect in WTA. This study provides proof for the robustness of the anchoring effect and neural evidence of the semantic priming model. Our findings indicate that activated contextual information, even seemingly irrelevant, can be embedded in the construction of economic value in the brain.

  2. Anchors as Semantic Primes in Value Construction: An EEG Study of the Anchoring Effect.

    Directory of Open Access Journals (Sweden)

    Qingguo Ma

    Full Text Available Previous research regarding anchoring effects has demonstrated that human judgments are often assimilated to irrelevant information. Studies have demonstrated that anchors influence the economic valuation of various products and experiences; however, the cognitive explanations of this effect remain controversial, and its neural mechanisms have rarely been explored. In the current study, we conducted an electroencephalography (EEG experiment to investigate the anchoring effect on willingness to accept (WTA for an aversive hedonic experience and the role of anchors in this judgment heuristic. The behavioral results demonstrated that random numbers affect participants' WTA for listening to pieces of noise. The participants asked for higher pay after comparing their WTA with higher numbers. The EEG results indicated that anchors also influenced the neural underpinnings of the valuation process. Specifically, when a higher anchor number was drawn, larger P2 and late positive potential amplitudes were elicited, reflecting the anticipation of more intensive pain from the subsequent noise. Moreover, higher anchors induced a stronger theta band power increase compared with lower anchors when subjects listened to the noises, indicating that the participants felt more unpleasant during the actual experience of the noise. The levels of unpleasantness during both anticipation and experience were consistent with the semantic information implied by the anchors. Therefore, these data suggest that a semantic priming process underlies the anchoring effect in WTA. This study provides proof for the robustness of the anchoring effect and neural evidence of the semantic priming model. Our findings indicate that activated contextual information, even seemingly irrelevant, can be embedded in the construction of economic value in the brain.

  3. Improvement in organophosphorus hydrolase activity of cell surface-engineered yeast strain using Flo1p anchor system.

    Science.gov (United States)

    Fukuda, Takeshi; Tsuchiyama, Kouta; Makishima, Hirokazu; Takayama, Katsumi; Mulchandani, Ashok; Kuroda, Kouichi; Ueda, Mitsuyoshi; Suye, Shin-ichiro

    2010-05-01

    Organophosphorus hydrolase (OPH) hydrolyzes organophosphorus esters. We constructed the yeast-displayed OPH using Flo1p anchor system. In this system, the N-terminal region of the protein was fused to Flo1p and the fusion protein was displayed on the cell surface. Hydrolytic reactions with paraoxon were carried out during 24 h of incubation of OPH-displaying cells at 30 degrees C. p-Nitrophenol produced in the reaction mixture was detected by HPLC. The strain with highest activity showed 8-fold greater OPH activity compared with cells engineered using glycosylphosphatidylinositol anchor system, and showed 20-fold greater activity than Escherichia coli using the ice nucleation protein anchor system. These results indicate that Flo1p anchor system is suitable for display of OPH in the cell surface-expression systems.

  4. Lash Transported Anchor for a Tanker Mooring.

    Science.gov (United States)

    1980-09-01

    including additional model tests, is needed. The anchor configurations presented in Figures llb and llc will function adequately for the ATTF on all...causeways at sea) and offshore oil companies (mating jacket type structures at sea). 4. A suitable anchor for rock would be twice as heavy (4,000 kips) as the...NIARINF FNVIRON. LAW .. IIAIVl’RN): Seattle "A (E. ianger): Sceittle. %%A.’ Itransportation, ( oust rUCtIo & CO1m IN %IR(.IN IA INST. OF- MARINE SOI

  5. Anchoring the Panic Disorder Severity Scale

    Science.gov (United States)

    Keough, Meghan E.; Porter, Eliora; Kredlow, M. Alexandra; Worthington, John J.; Hoge, Elizabeth A.; Pollack, Mark H.; Shear, M. Katherine; Simon, Naomi M.

    2012-01-01

    The Panic Disorder Severity Scale (PDSS) is a clinician-administered measure of panic disorder symptom severity widely used in clinical research. This investigation sought to provide clinically meaningful anchor points for the PDSS both in terms of clinical severity as measured by the Clinical Global Impression-Severity Scale (CGI-S) and to extend…

  6. The bone-anchored hearing aid

    DEFF Research Database (Denmark)

    Foghsgaard, Søren

    2014-01-01

    The bone-anchored hearing aid (Baha) was introduced in 1977 by Tjellström and colleagues and has now been used clinically for over 30 years. Generally, the outcomes are good, and several studies have shown improved audiological- and quality of life outcomes. The principle of the Baha is, that sound...

  7. Multimodality of Learning Through Anchored Instruction

    Science.gov (United States)

    Love, Mary Susan

    2004-01-01

    Multimodality of learning results from the intertextual relationship between multimodal design and other meaning-making modes. Meaning making is becoming more multimodal because language is continually reshaped by new forms of communication media. This article examines anchored instruction from a multimodal perspective. The first section includes…

  8. Effects of Media Attributes in Anchored Instruction.

    Science.gov (United States)

    Shyu, Hsin-Yih

    1999-01-01

    Investigates the effects of computer-assisted video-based anchored instruction on promoting students' attitudes toward mathematical instruction and problem-solving skills. Examines the effects of different media attributes on students' mathematical achievement and attitudes in a situated learning environment. Findings suggest that anchored…

  9. Anchor Stress Checking of Security Injection Tank

    Institute of Scientific and Technical Information of China (English)

    2011-01-01

    The intention of the calculating is to check the anchor stresses of the security injection tank to know whether the stress is satisfied the code requirements on the basis of all the reaction forces gained in the static, seismic and thermal stress results.

  10. The "Anchor" Method: Principle and Practice.

    Science.gov (United States)

    Selgin, Paul

    This report discusses the "anchor" language learning method that is based upon derivation rather than construction, using Italian as an example of a language to be learned. This method borrows from the natural process of language learning as it asks the student to remember whole expressions that serve as vehicles for learning both words and rules,…

  11. Fouled Anchors: The CONSTELLATION Question Answered

    Science.gov (United States)

    1991-09-01

    1711804 87AU 73 89-1-3401 11. TITLE (kcn e S.cE) Caut,.cawn) Fouled Anchors: The Constellation Question Answered IL. PERSONA .. AUTHOPS) Wegner, Dana M...was familia , with the unaltered Constellation in Newport and, unknown to the Committee, had indeed visited the ship once in Baltimore (see p. 45). It is

  12. Predicting Anchor Links between Heterogeneous Social Networks

    CERN Document Server

    Sajadmanesh, Sina; Khodadadi, Ali

    2016-01-01

    People usually get involved in multiple social networks to enjoy new services or to fulfill their needs. Many new social networks try to attract users of other existing networks to increase the number of their users. Once a user (called source user) of a social network (called source network) joins a new social network (called target network), a new inter-network link (called anchor link) is formed between the source and target networks. In this paper, we concentrated on predicting the formation of such anchor links between heterogeneous social networks. Unlike conventional link prediction problems in which the formation of a link between two existing users within a single network is predicted, in anchor link prediction, the target user is missing and will be added to the target network once the anchor link is created. To solve this problem, we use meta-paths as a powerful tool for utilizing heterogeneous information in both the source and target networks. To this end, we propose an effective general meta-pat...

  13. Finding Chemical Anchors in the Kitchen

    Science.gov (United States)

    Haim, Liliana

    2005-01-01

    ''The Chemistry Kitchen'', a unit composed of five activities with kitchen elements for elementary students ages 9-11, introduces the children to the skills and chemical working ideas to be used later as anchors for chemical concepts. These activities include kitchen elements, determining the relative mass and so on.

  14. Stone anchors of India: Findings, classification and significance.

    Digital Repository Service at National Institute of Oceanography (India)

    Tripati, S.

    and size do not resemble the anchors found during maritime archaeological explorations since 1986. The stone anchors until now recovered from many sites of India can broadly be classified into four types: namely composite, Indo-Arabian, ring stone (mushroom...

  15. Membrane anchor R9AP potentiates GTPase-accelerating protein activity of RGS11 x Gbeta5 complex and accelerates inactivation of the mGluR6-G(o) signaling.

    Science.gov (United States)

    Masuho, Ikuo; Celver, Jeremy; Kovoor, Abraham; Martemyanov, Kirill A

    2010-02-12

    The R7 subfamily of RGS proteins critically regulates neuronal G protein-signaling pathways that are essential for vision, nociception, motor coordination, and reward processing. A member of the R7 RGS family, RGS11, is a GTPase-accelerating protein specifically expressed in retinal ON-bipolar cells where it forms complexes with the atypical G protein beta subunit, Gbeta(5), and transmembrane protein R9AP. Association with R9AP has been shown to be critical for the proteolytic stability of the complex in the retina. In this study we report that R9AP can in addition stimulate the GTPase-accelerating protein activity of the RGS11 x Gbeta(5) complex at Galpha(o). Single turnover GTPase assays reveal that R9AP co-localizes RGS11 x Gbeta(5) and Galpha(o) on the membrane and allosterically potentiates the GTPase-accelerating function of RGS11 x Gbeta(5). Reconstitution of mGluR6-Galpha(o) signaling in Xenopus oocytes indicates that RGS11 x Gbeta(5)-mediated GTPase acceleration in this system requires co-expression of R9AP. The results provide new insight into the regulation of mGluR6-Galpha(o) signaling by the RGS11 x Gbeta(5) x R9AP complex and establish R9AP as a general GTPase-accelerating protein activity regulator of R7 RGS complexes.

  16. FOS-1 promotes basement-membrane removal during anchor-cell invasion in C. elegans.

    Science.gov (United States)

    Sherwood, David R; Butler, James A; Kramer, James M; Sternberg, Paul W

    2005-06-17

    Cell invasion through basement membranes is crucial during morphogenesis and cancer metastasis. Here, we genetically dissect this process during anchor-cell invasion into the vulval epithelium in C. elegans. We have identified the fos transcription factor ortholog fos-1 as a critical regulator of basement-membrane removal. In fos-1 mutants, the gonadal anchor cell extends cellular processes normally toward vulval cells, but these processes fail to remove the basement membranes separating the gonad from the vulval epithelium. fos-1 is expressed in the anchor cell and controls invasion cell autonomously. We have identified ZMP-1, a membrane-type matrix metalloproteinase, CDH-3, a Fat-like protocadherin, and hemicentin, a fibulin family extracellular matrix protein, as transcriptional targets of FOS-1 that promote invasion. These results reveal a key genetic network that controls basement-membrane removal during cell invasion.

  17. Anchored Instruction and Its Relationship to Situated Cognition.

    Science.gov (United States)

    Educational Researcher, 1990

    1990-01-01

    Discusses anchored instruction, a teaching technique using videodisc-based problem-solving environments, and describes two research projects involving anchored instruction. Argues that anchored instruction provides a way to recreate and improve upon some of the advantages of situated cognition. (FMW)

  18. Anchored Instruction for Chinese Students: Enhancing Attitudes toward Mathematics.

    Science.gov (United States)

    Shyu, Hsin-Yih

    1997-01-01

    The purpose of this study was to develop a videodisc-based computer-aided multimedia-anchored instruction for Chinese students and to examine its effects on student attitudes toward mathematics and anchored instruction. Participants were 74 fifth graders. Results indicated that anchored instruction improved students' attitudes toward mathematics…

  19. Students' Anchoring Predisposition: An Illustration from Spring Training Baseball

    Science.gov (United States)

    Mohrweis, Lawrence C.

    2014-01-01

    The anchoring tendency results when decision makers anchor on initial values and then make final assessments that are adjusted insufficiently away from the initial values. The professional literature recognizes that auditors often risk falling into the judgment trap of anchoring and adjusting (Ranzilla et al., 2011). Students may also be unaware…

  20. GUP1 of Saccharomyces cerevisiae encodes an O-acyltransferase involved in remodeling of the GPI anchor.

    Science.gov (United States)

    Bosson, Régine; Jaquenoud, Malika; Conzelmann, Andreas

    2006-06-01

    The anchors of mature glycosylphosphatidylinositol (GPI)-anchored proteins of Saccharomyces cerevisiae contain either ceramide or diacylglycerol with a C26:0 fatty acid in the sn2 position. The primary GPI lipid added to newly synthesized proteins in the ER consists of diacylglycerol with conventional C16 and C18 fatty acids. Here we show that GUP1 is essential for the synthesis of the C26:0-containing diacylglycerol anchors. Gup1p is an ER membrane protein with multiple membrane-spanning domains harboring a motif that is characteristic of membrane-bound O-acyl-transferases (MBOAT). Gup1Delta cells make normal amounts of GPI proteins but most mature GPI anchors contain lyso-phosphatidylinositol, and others possess phosphatidylinositol with conventional C16 and C18 fatty acids. The incorporation of the normal ceramides into the anchors is also disturbed. As a consequence, the ER-to-Golgi transport of the GPI protein Gas1p is slow, and mature Gas1p is lost from the plasma membrane into the medium. Gup1Delta cells have fragile cell walls and a defect in bipolar bud site selection. GUP1 function depends on the active site histidine of the MBOAT motif. GUP1 is highly conserved among fungi and protozoa and the gup1Delta phenotype is partially corrected by GUP1 homologues of Aspergillus fumigatus and Trypanosoma cruzi.

  1. Molecular interactions of the neuronal GPI-anchored lipocalin Lazarillo.

    Science.gov (United States)

    Sanchez, Diego; Ortega-Cubero, Sara; Akerström, Bo; Herrera, Macarena; Bastiani, Michael J; Ganfornina, Maria D

    2008-01-01

    Lazarillo, a glycoprotein involved in axon growth and guidance in the grasshopper embryo, is the only member of the lipocalin family that is attached to the cell surface by a GPI anchor. Recently, the study of Lazarillo homologous genes in Drosophila and mouse has revealed new functions in the regulation of lifespan, stress resistance and neurodegeneration. Here we report an analysis of biochemical properties of Lazarillo to gain insight into the molecular basis of its physiological function. Recombinant forms of the grasshopper protein were expressed in two different systems to test: (1) potential binding of several hydrophobic ligands; (2) protein-protein homophilic interactions; and (3) whether interaction with the function-blocking mAb 10E6 interferes with ligand binding. We tested 10 candidate ligands (retinoic acid, heme, bilirubin, biliverdin, ecdysterone, juvenile hormone, farnesol, arachidonic acid, linoleic acid and palmitic acid), and monitored binding using electrophoretic mobility shift, absorbance spectrum, and fluorimetry assays. Our work indicates binding to heme and retinoic acid, resulting in increased electrophoretic mobility, as well as to fatty acids, resulting in multimerization. Retinoic acid and fatty acids binding were confirmed by fluorescence titration, and heme binding was confirmed with absorbance spectrum assays. We demonstrate that Lazarillo oligomerizes in solution and can form clusters in the plasma membrane when expressed and GPI-anchored to the cell surface, however it is unable to mediate cell-cell adhesion. Finally, by ligand-mAb competition experiments we show that ligand-binding alone cannot be the key factor for Lazarillo to perform its function during axonal growth in the grasshopper embryo.

  2. Composite materials formed with anchored nanostructures

    Energy Technology Data Exchange (ETDEWEB)

    Seals, Roland D; Menchhofer, Paul A; Howe, Jane Y; Wang, Wei

    2015-03-10

    A method of forming nano-structure composite materials that have a binder material and a nanostructure fiber material is described. A precursor material may be formed using a mixture of at least one metal powder and anchored nanostructure materials. The metal powder mixture may be (a) Ni powder and (b) NiAl powder. The anchored nanostructure materials may comprise (i) NiAl powder as a support material and (ii) carbon nanotubes attached to nanoparticles adjacent to a surface of the support material. The process of forming nano-structure composite materials typically involves sintering the mixture under vacuum in a die. When Ni and NiAl are used in the metal powder mixture Ni.sub.3Al may form as the binder material after sintering. The mixture is sintered until it consolidates to form the nano-structure composite material.

  3. Electrochromic mirror using viologen-anchored nanoparticles

    Energy Technology Data Exchange (ETDEWEB)

    Kim, Han Na [Electronics and Telecommunications Research Institute, Nature-mimic I/O interface Research Section, 218 Gajeong-roYuseong-gu, Daejeon 305-700 (Korea, Republic of); University of Science and Technology, Advanced Device Technology, 217 Gajeong-roYuseong-gu, Daejeon 305-350 (Korea, Republic of); Cho, Seong M.; Ah, Chil Seong; Song, Juhee; Ryu, Hojun; Kim, Yong Hae [Electronics and Telecommunications Research Institute, Nature-mimic I/O interface Research Section, 218 Gajeong-roYuseong-gu, Daejeon 305-700 (Korea, Republic of); Kim, Tae-Youb, E-mail: youby@etri.re.kr [Electronics and Telecommunications Research Institute, Nature-mimic I/O interface Research Section, 218 Gajeong-roYuseong-gu, Daejeon 305-700 (Korea, Republic of); University of Science and Technology, Advanced Device Technology, 217 Gajeong-roYuseong-gu, Daejeon 305-350 (Korea, Republic of)

    2016-10-15

    Highlights: • Three types of ECM device were fabricated using viologen-anchored ECDs. • The devices were investigated according to their optical structures. • The anti-reflection material affects the reflectance and the coloration efficiency. • The device design of ECMs is a crucial factor for clear reflected images. - Abstract: Electrochromic mirrors (ECMs) that are used in automobile mirrors need to have high reflectance, a high contrast ratio, and a clear image. In particular, it is critical that distortions of clear images are minimized for safety. Therefore, an ECM is fabricated using viologen-anchored nanoparticles and a magnesium fluoride (MgF{sub 2}) layer with an anti-reflection function. The ECM has approximately 30.42% in the reflectance dynamic range and 125 cm{sup 2}/C high coloration efficiency.

  4. An Analytical Method for Positioning Drag Anchors in Seabed Soils

    Institute of Scientific and Technical Information of China (English)

    张炜; 刘海笑; 李新仲; 李清平; 曹静

    2015-01-01

    Positioning drag anchors in seabed soils are strongly influenced not only by the properties of the anchor and soil, but also by the characteristics of the installation line. The investigation on the previous prediction methods related to anchor positioning demonstrates that the prediction of the anchor position during dragging has inevitably introduced some key and unsubstantiated hypotheses and the applicability of these methods is limited. In the present study, the interactional system between the drag anchor and installation line is firstly introduced for the analysis of anchor positioning. Based on the two mechanical models for embedded lines and drag anchors, the positioning equations for drag anchors have been derived both for cohesive and noncohesive soils. Since the drag angle at the shackle is the most important parameter in the positioning equations, a novel analytical method that can predict both the variation and the exact value of the drag angle at the shackle is proposed. The analytical method for positioning drag anchors which combines the interactional system between the drag anchor and the installation line has provided a reasonable theoretic approach to investigate the anchor behaviors in soils. By comparing with the model flume experiments, the sensitivity, effectiveness and veracity of the positioning method are well verified.

  5. Anchored Instruction: Situiertes Lernen in multimedialen Lernumgebungen

    OpenAIRE

    2001-01-01

    «Anchored Instruction» ist ein konstruktivistischer Ansatz aus den USA, dessen Vertreter technologisch unterstützte, situierte Lernumgebungen für verschiedene schulische Fächer und Altersstufen entwickeln, implementieren und erforschen. Wichtige Ziele, Gestaltungsprinzipien und Merkmale solcher Lernumgebungen sowie ihre Implementation im Unterricht werden anhand von zwei Anwendungen näher dargestellt: (1) «The Adventures of Jasper Woodbury», eine Serie von Video-Geschichten und Zusatzmaterial...

  6. Developing an "anchor" system to enhance postural control.

    Science.gov (United States)

    Mauerberg-deCastro, Eliane

    2004-07-01

    This article uses an anchor metaphor to explain the dynamic interplay between the human body's active uses of nonrigid tools to mediate information about its adjacent environment to enhance postural control. The author used an "anchor" system (ropes attached to varying weights resting on the floor) to test blindfolded adults who performed a restricted-balance task (30 s one-foot standing). Participants were tested while holding the anchors under a variety of weight conditions (125 g, 250 g, 500 g, and 1 kg) and again during a baseline condition (no anchors). When compared with the baseline condition, there was a significant reduction in the amount of body sway across the anchor conditions. The author found that mechanical support provided by the anchor system was secondary to its haptic exploratory function and that an individual can use the anchoring strategy with a dual purpose: for resting and for reorientation after intrinsic disruptions.

  7. Anchoring properties of substrate with a grating surface

    Institute of Scientific and Technical Information of China (English)

    Ye Wen-Jiang; Xing Hong-Yu; Yang Guo-Chen

    2007-01-01

    The anchoring properties of substrate with a grating surface are investigated analytically. The alignment of nematic liquid crystal (NLC) in a grating surface originates from two mechanisms, thus the anchoring energy consists of two parts. One originates from the interaction potential between NLC molecules and the molecules on the substrate surface,and the other stems from the increased elastic strain energy. Based on the two mechanisms, the expression of anchoring energy per unit area of a projected plane of this grating surface is deduced and called the equivalent anchoring energy formula. Both the strength and the easy direction of equivalent anchoring energy are a function of the geometrical parameters (amplitude and pitch) of a grating surface. By using this formula, the grating surface can be replaced by its projected plane and its anchoring properties can be described by the equivalent anchoring energy formula.

  8. Liquid-phase synthesis of bridged peptides using olefin metathesis of a protected peptide with a long aliphatic chain anchor.

    Science.gov (United States)

    Aihara, Keisuke; Komiya, Chiaki; Shigenaga, Akira; Inokuma, Tsubasa; Takahashi, Daisuke; Otaka, Akira

    2015-02-01

    Bridged peptides including stapled peptides are attractive tools for regulating protein-protein interactions (PPIs). An effective synthetic methodology in a heterogeneous system for the preparation of these peptides using olefin metathesis and hydrogenation of protected peptides with a long aliphatic chain anchor is reported.

  9. SwissProt search result: AK106271 [KOME

    Lifescience Database Archive (English)

    Full Text Available AK106271 002-100-G09 (Q28628) A-kinase anchor protein 9 (Protein kinase A anchoring... protein 9) (PRKA9) (A-kinase anchor protein 120 kDa) (AKAP 120) (Fragment) AKAP9_RABIT 2e-12 ...

  10. SwissProt search result: AK063431 [KOME

    Lifescience Database Archive (English)

    Full Text Available AK063431 001-115-D10 (Q28628) A-kinase anchor protein 9 (Protein kinase A anchoring... protein 9) (PRKA9) (A-kinase anchor protein 120 kDa) (AKAP 120) (Fragment) AKAP9_RABIT 1e-11 ...

  11. Robotic Ankle for Omnidirectional Rock Anchors

    Science.gov (United States)

    Parness, Aaron; Frost, Matthew; Thatte, Nitish

    2013-01-01

    Future robotic exploration of near-Earth asteroids and the vertical and inverted rock walls of lava caves and cliff faces on Mars and other planetary bodies would require a method of gripping their rocky surfaces to allow mobility without gravitational assistance. In order to successfully navigate this terrain and drill for samples, the grippers must be able to produce anchoring forces in excess of 100 N. Additionally, the grippers must be able to support the inertial forces of a moving robot, as well gravitational forces for demonstrations on Earth. One possible solution would be to use microspine arrays to anchor to rock surfaces and provide the necessary load-bearing abilities for robotic exploration of asteroids. Microspine arrays comprise dozens of small steel hooks supported on individual suspensions. When these arrays are dragged along a rock surface, the steel hooks engage with asperities and holes on the surface. The suspensions allow for individual hooks to engage with asperities while the remaining hooks continue to drag along the surface. This ensures that the maximum possible number of hooks engage with the surface, thereby increasing the load-bearing abilities of the gripper. Using the microspine array grippers described above as the end-effectors of a robot would allow it to traverse terrain previously unreachable by traditional wheeled robots. Furthermore, microspine-gripping robots that can perch on cliffs or rocky walls could enable a new class of persistent surveillance devices for military applications. In order to interface these microspine grippers with a legged robot, an ankle is needed that can robotically actuate the gripper, as well as allow it to conform to the large-scale irregularities in the rock. The anchor serves three main purposes: deploy and release the anchor, conform to roughness or misalignment with the surface, and cancel out any moments about the anchor that could cause unintentional detachment. The ankle design contains a

  12. Dynamic performance of concrete undercut anchors for Nuclear Power Plants

    Energy Technology Data Exchange (ETDEWEB)

    Mahrenholtz, Christoph, E-mail: christoph@mahrenholtz.net; Eligehausen, Rolf

    2013-12-15

    Graphical abstract: - Highlights: • Behavior of undercut anchors under dynamic actions simulating earthquakes. • First high frequency load and crack cycling tests on installed concrete anchors ever. • Comprehensive review of anchor qualification for Nuclear Power Plants. - Abstract: Post-installed anchors are widely used for structural and nonstructural connections to concrete. In many countries, concrete anchors used for Nuclear Power Plants have to be qualified to ensure reliable behavior even under extreme conditions. The tests required for qualification of concrete anchors are carried out at quasi-static loading rates well below the rates to be expected for dynamic actions deriving from earthquakes, airplane impacts or explosions. To investigate potentially beneficial effects of high loading rates and cycling frequencies, performance tests on installed undercut anchors were conducted. After introductory notes on anchor technology and a comprehensive literature review, this paper discusses the qualification of anchors for Nuclear Power Plants and the testing carried out to quantify experimentally the effects of dynamic actions on the load–displacement behavior of undercut anchors.

  13. Aurora A kinase activates YAP signaling in triple-negative breast cancer.

    Science.gov (United States)

    Chang, S-S; Yamaguchi, H; Xia, W; Lim, S-O; Khotskaya, Y; Wu, Y; Chang, W-C; Liu, Q; Hung, M-C

    2017-03-02

    The Yes-associated protein (YAP) is an effector that transduces the output of the Hippo pathway to transcriptional modulation. Considering the role of YAP in cancers, this protein has emerged as a key node in malignancy development. In this study, we determined that Aurora A kinase acts as a positive regulator for YAP-mediated transcriptional machinery. Specifically, YAP associates with Aurora A predominantly in the nucleus. Activation of Aurora A can impinge on YAP activity through direct phosphorylation. Moreover, aberrant expression of YAP and Aurora A signaling is highly correlated with triple-negative breast cancer (TNBC). We herein provide evidence to establish the functional relevance of this newly discovered regulatory axis in TNBC.

  14. Binding equilibrium and kinetics of membrane-anchored receptors and ligands in cell adhesion: Insights from computational model systems and theory.

    Science.gov (United States)

    Weikl, Thomas R; Hu, Jinglei; Xu, Guang-Kui; Lipowsky, Reinhard

    2016-09-02

    The adhesion of cell membranes is mediated by the binding of membrane-anchored receptor and ligand proteins. In this article, we review recent results from simulations and theory that lead to novel insights on how the binding equilibrium and kinetics of these proteins is affected by the membranes and by the membrane anchoring and molecular properties of the proteins. Simulations and theory both indicate that the binding equilibrium constant [Formula: see text] and the on- and off-rate constants of anchored receptors and ligands in their 2-dimensional (2D) membrane environment strongly depend on the membrane roughness from thermally excited shape fluctuations on nanoscales. Recent theory corroborated by simulations provides a general relation between [Formula: see text] and the binding constant [Formula: see text] of soluble variants of the receptors and ligands that lack the membrane anchors and are free to diffuse in 3 dimensions (3D).

  15. ANCHORING IN THE POLITICAL AND ECONOMIC MARKETING

    Directory of Open Access Journals (Sweden)

    Tatyana L. Shklyar

    2015-01-01

    Full Text Available The main purpose of marketing, both ineconomy and in politics is to attract themaximum number of customers or voters, maximizing customer satisfaction and ,ideally, improve the quality of life.The author, in various aspects, thetechnology of anchoring used in NLP, to attract customers and voters, both in the economy and in politics.In different examples demonstrate theoverall impact on the psychology of the consumer. Separating policy and the economy, marketers are missing something. The author proposes to look at how psychologicalanchors affect these two, at fi rst glance, different vector.

  16. Adsorption phenomena and anchoring energy in nematic liquid crystals

    CERN Document Server

    Barbero, Giovanni

    2005-01-01

    Despite the large quantity of phenomenological information concerning the bulk properties of nematic phase liquid crystals, little is understood about the origin of the surface energy, particularly the surface, interfacial, and anchoring properties of liquid crystals that affect the performance of liquid crystal devices. Self-contained and unique, Adsorption Phenomena and Anchoring Energy in Nematic Liquid Crystals provides an account of new and established results spanning three decades of research into the problems of anchoring energy and adsorption phenomena in liquid crystals.The book contains a detailed discussion of the origin and possible sources of anchoring energy in nematic liquid crystals, emphasizing the dielectric contribution to the anchoring energy in particular. Beginning with fundamental surface and anchoring properties of liquid crystals and the definition of the nematic phase, the authors explain how selective ion adsorption, dielectric energy density, thickness dependence, and bias voltage...

  17. PAQR3 modulates cholesterol homeostasis by anchoring Scap/SREBP complex to the Golgi apparatus.

    Science.gov (United States)

    Xu, Daqian; Wang, Zheng; Zhang, Yuxue; Jiang, Wei; Pan, Yi; Song, Bao-Liang; Chen, Yan

    2015-08-27

    Cholesterol biosynthesis is regulated by transcription factors SREBPs and their escort protein Scap. On sterol depletion, Scap/SREBP complex is transported from endoplasmic reticulum (ER) to the Golgi apparatus where SREBP is activated. Under cholesterol sufficient condition, Insigs act as anchor proteins to retain Scap/SREBP in the ER. However, the anchor protein of Scap/SREBP in the Golgi is unknown. Here we report that a Golgi-localized membrane protein progestin and adipoQ receptors 3 (PAQR3) interacts with Scap and SREBP and tethers them to the Golgi. PAQR3 promotes Scap/SREBP complex formation, potentiates SREBP processing and enhances lipid synthesis. The mutually exclusive interaction between Scap and PAQR3 or Insig-1 is regulated by cholesterol level. PAQR3 knockdown in liver blunts SREBP pathway and decreases hepatic cholesterol content. Disrupting the interaction of PAQR3 with Scap/SREBP by a synthetic peptide inhibits SREBP processing and activation. Thus, PAQR3 regulates cholesterol homeostasis by anchoring Scap/SREBP to the Golgi and disruption of such function reduces cholesterol biosynthesis.

  18. Allosteric inhibition of Aurora-A kinase by a synthetic vNAR domain.

    Science.gov (United States)

    Burgess, Selena G; Oleksy, Arkadiusz; Cavazza, Tommaso; Richards, Mark W; Vernos, Isabelle; Matthews, David; Bayliss, Richard

    2016-07-01

    The vast majority of clinically approved protein kinase inhibitors target the ATP-binding pocket directly. Consequently, many inhibitors have broad selectivity profiles and most have significant off-target effects. Allosteric inhibitors are generally more selective, but are difficult to identify because allosteric binding sites are often unknown or poorly characterized. Aurora-A is activated through binding of TPX2 to an allosteric site on the kinase catalytic domain, and this knowledge could be exploited to generate an inhibitor. Here, we generated an allosteric inhibitor of Aurora-A kinase based on a synthetic, vNAR single domain scaffold, vNAR-D01. Biochemical studies and a crystal structure of the Aurora-A/vNAR-D01 complex show that the vNAR domain overlaps with the TPX2 binding site. In contrast with the binding of TPX2, which stabilizes an active conformation of the kinase, binding of the vNAR domain stabilizes an inactive conformation, in which the αC-helix is distorted, the canonical Lys-Glu salt bridge is broken and the regulatory (R-) spine is disrupted by an additional hydrophobic side chain from the activation loop. These studies illustrate how single domain antibodies can be used to characterize the regulatory mechanisms of kinases and provide a rational basis for structure-guided design of allosteric Aurora-A kinase inhibitors.

  19. Influence of Anchoring on Burial Depth of Submarine Pipelines

    OpenAIRE

    Yuan Zhuang; Yang Li; Wei Su

    2016-01-01

    Since the beginning of the twenty-first century, there has been widespread construction of submarine oil-gas transmission pipelines due to an increase in offshore oil exploration. Vessel anchoring operations are causing more damage to submarine pipelines due to shipping transportation also increasing. Therefore, it is essential that the influence of anchoring on the required burial depth of submarine pipelines is determined. In this paper, mathematical models for ordinary anchoring and emerge...

  20. Anchor-induced chondral damage in the hip.

    Science.gov (United States)

    Matsuda, Dean K; Bharam, Srino; White, Brian J; Matsuda, Nicole A; Safran, Marc

    2015-01-01

    The purpose of this study is to investigate the outcomes from anchor-induced chondral damage of the hip, both with and without frank chondral penetration. A multicenter retrospective case series was performed of patients with chondral deformation or penetration during initial hip arthroscopic surgery. Intra-operative findings, post-surgical clinical courses, hip outcome scores and descriptions of arthroscopic treatment in cases requiring revision surgery and anchor removal are reported. Five patients (three females) of mean age 32 years (range, 16-41 years) had documented anchor-induced chondral damage with mean 3.5 years (range, 1.5-6.0 years) follow-up. The 1 o'clock position (four cases) and anterior and mid-anterior portals (two cases each) were most commonly implicated. Two cases of anchor-induced acetabular chondral deformation without frank penetration had successful clinical and radiographic outcomes, while one case progressed from deformation to chondral penetration with clinical worsening. Of the cases that underwent revision hip arthroscopy, all three had confirmed exposed hard anchors which were removed. Two patients have had clinical improvement and one patient underwent early total hip arthroplasty. Anchor-induced chondral deformation without frank chondral penetration may be treated with close clinical and radiographic monitoring with a low threshold for revision surgery and anchor removal. Chondral penetration should be treated with immediate removal of offending hard anchor implants. Preventative measures include distal-based portals, small diameter and short anchors, removable hard anchors, soft suture-based anchors, curved drill and anchor insertion instrumentation and attention to safe trajectories while visualizing the acetabular articular surface.

  1. Anchor-induced chondral damage in the hip

    Science.gov (United States)

    Matsuda, Dean K.; Bharam, Srino; White, Brian J.; Matsuda, Nicole A.; Safran, Marc

    2015-01-01

    The purpose of this study is to investigate the outcomes from anchor-induced chondral damage of the hip, both with and without frank chondral penetration. A multicenter retrospective case series was performed of patients with chondral deformation or penetration during initial hip arthroscopic surgery. Intra-operative findings, post-surgical clinical courses, hip outcome scores and descriptions of arthroscopic treatment in cases requiring revision surgery and anchor removal are reported. Five patients (three females) of mean age 32 years (range, 16–41 years) had documented anchor-induced chondral damage with mean 3.5 years (range, 1.5–6.0 years) follow-up. The 1 o'clock position (four cases) and anterior and mid-anterior portals (two cases each) were most commonly implicated. Two cases of anchor-induced acetabular chondral deformation without frank penetration had successful clinical and radiographic outcomes, while one case progressed from deformation to chondral penetration with clinical worsening. Of the cases that underwent revision hip arthroscopy, all three had confirmed exposed hard anchors which were removed. Two patients have had clinical improvement and one patient underwent early total hip arthroplasty. Anchor-induced chondral deformation without frank chondral penetration may be treated with close clinical and radiographic monitoring with a low threshold for revision surgery and anchor removal. Chondral penetration should be treated with immediate removal of offending hard anchor implants. Preventative measures include distal-based portals, small diameter and short anchors, removable hard anchors, soft suture-based anchors, curved drill and anchor insertion instrumentation and attention to safe trajectories while visualizing the acetabular articular surface. PMID:27011815

  2. AUV Load Separation Motion with Constraint of Anchor Chain

    Institute of Scientific and Technical Information of China (English)

    SHAO Cheng; SONG Bao-wei; DU Xiao-xu; WANG Peng; LI Jia-wang

    2009-01-01

    Motion equations of AUV(autonomous underwater vehicle) load separation with the constraint of anchor chain is derived. Based on proper engineering assumptions for anchor chain,system viewpoint is used to found the motion equations, and the D'Alembert principle is used to eliminate the constraint force of anchor chain. Based on the equations, the motion simulation is carried out to a certain AUV, which reflects the actual condition, and is used for the reference of resrarching AUV load separation motion with the constraint of anchor chain.

  3. Infrastructure anchor bolt inspection program with NDE applications

    Science.gov (United States)

    Fish, Philip E.

    1996-11-01

    In 1990, Wisconsin Department of Transportation found a high mast light pole with two of six anchor bolts failed. This failure along with published reports from Michigan DOT about anchor bolt failures on cantilever sign structures, raised concern about the quality and condition of anchor bolts on the Wisconsin DOT system. Wisconsin Department of Transportation implemented an Anchor Bolt Inspection Program in 1990 for cantilever sign structures, high mast light towers, interstate light towers, and signal masts. The program requires an experienced inspection team and a practical inspection approach. Inspection preparation includes review of all background information such as design plans, design computations, construction plans, shop plans, and maintenance history. An inspection plan is developed. Special emphasis is placed on determining material type, cut or rolled threads, and type of coating for anchor bolts. Inspection emphasis are on "hands on" and Nondestructive evaluation. Special emphasis is placed on visual conditions of anchor bolts (cut or rolled threads, straightness, corrosion, nut tension etc.) along with ultrasonic inspection. This program places a strong emphasis on Non Destructive Testing (NDT), especially ultrasonic. Procedures and inspection calibrations are developed from similar anchor bolt geometry and material type. Cut notches are placed in the anchor bolts at locations of possible failure. NDT inspection calibrations are performed from these bolts. Report documentation includes all design plans, pictorial documentation of structural deficiencies, sketches, nondestructive evaluation reports, conclusions, and recommendations. This program has been successful in locating failed anchor bolts and critical cracks before failure of an entire structure.

  4. Calculation of prestressed anchor segment by 3D infiniteelement

    Institute of Scientific and Technical Information of China (English)

    Yanfen WANG; Hongyang XIE; Yuanhan WANG

    2009-01-01

    Based on 1D infinite element theory, the coordinate transformation and shape function of 3D point-radiation 4-node infinite elements were derived.They were coupled with 8-node finite elements to compute the compressive deformation of the prestressed anchor segment. The results indicate that when the prestressed force acts on the anchor segment, the stresses and displacements in the rock around the anchor segment are concentrated in the zone center with the anchor axis and are subjected to exponential decay. Therefore, the stresses and the displacement spindles are formed. The calculation results of the infinite element are close to the theoretical results.

  5. Phosphatidylinositol anchor of HeLa cell alkaline phosphatase

    Energy Technology Data Exchange (ETDEWEB)

    Jemmerson, R.; Low, M.G.

    1987-09-08

    Alkaline phosphatase from cancer cells, HeLa TCRC-1, was biosynthetically labeled with either /sup 3/H-fatty acids or (/sup 3/H)ethanolamine as analyzed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and fluorography of immunoprecipitated material. Phosphatidylinositol-specific phospholipase C (PI-PLC) released a substantial proportion of the /sup 3/H-fatty acid label from immunoaffinity-purified alkaline phosphatase but had no effect on the radioactivity of (/sup 3/H)ethanolamine-labeled material. PI-PLC also liberated catalytically active alkaline phosphatase from viable cells, and this could be selectively blocked by monoclonal antibodies to alkaline phosphatase. However, the alkaline phosphatase released from /sup 3/H-fatty acid labeled cells by PI-PLC was not radioactive. By contrast, treatment with bromelain removed both the /sup 3/H-fatty acid and the (/sup 3/H)ethanolamine label from purified alkaline phosphatase. Subtilisin was also able to remove the (/sup 3/H)ethanolamine label from the purified alkaline phosphatase. The /sup 3/H radioactivity in alkaline phosphatase purified from (/sup 3/H)ethanolamine-labeled cells comigrated with authentic (/sup 3/H)ethanolamine by anion-exchange chromatography after acid hydrolysis. The data suggest that the /sup 3/H-fatty acid and (/sup 3/H)ethanolamine are covalently attached to the carboxyl-terminal segment since bromelain and subtilisin both release alkaline phosphatase from the membrane by cleavage at that end of the polypeptide chain. The data are consistent with findings for other proteins recently shown to be anchored in the membrane through a glycosylphosphatidylinositol structure and indicate that a similar structure contributes to the membrane anchoring of alkaline phosphatase.

  6. Construction of a Pichia pastoris cell-surface display system using Flo1p anchor system.

    Science.gov (United States)

    Tanino, Takanori; Fukuda, Hideki; Kondo, Akihiko

    2006-01-01

    A Pichia pastoris cell-surface display system was constructed using a Flo1p anchor system, which was developed in Saccharomyces cerevisiae. The lipase from Rhizopus oryzae with a pro sequence (ProROL) was used as the model protein and was genetically fused to the anchor consisting of amino acids 1-1099 of Flo1p (FS anchor). The resulting fusion protein FSProROL was expressed under the control of the alcohol oxidase 1 promoter (pAOX1). The fluorescence microscopy of immunolabeled P. pastoris cells revealed that ProROL was displayed on the cell surface, and Western blot analysis revealed that the fusion protein FSProROL was noncovalently attached to the cell wall and highly glycosylated. The lipase activity of P. pastoris cells was affected by the methanol concentration for the induction phase. Surprisingly, the activity of lipase displayed on the cells incubated at 60 degrees C was not only stable but also increased to about 6.5 times the initial value after 4 h incubation.

  7. Anchored Instruction in Preservice Educational Technology Classes: A Research Project.

    Science.gov (United States)

    Bauer, Jeffrey W.

    This study examined the effectiveness of the anchored instruction (i.e., using a theme or anchor around which various learning activities take place) approach in preparing preservice teachers to integrate technology. Participants were 48 students enrolled in the three sections of a preservice educational technology class during the summer of 1997.…

  8. Effects of Knowledge Abstraction with Anchored Instruction on Learning Transfer.

    Science.gov (United States)

    Shih, Yu-Fen; And Others

    Transfer of learning is one of the major purposes of education. Theories and research have tried hard to answer questions such as: how does transfer occur? and how is transfer enhanced? Situated cognitive theory and research about anchored instruction together bring some positive findings. Anchored instruction provides the learner with a situated,…

  9. Using Anchored Instruction to Teach about Assistive Technology.

    Science.gov (United States)

    Blackhurst, A. Edward; Morse, Timothy E.

    1996-01-01

    A training module about assistive technology using an integrated hypermedia format and principles of anchored instruction was developed and field tested with 57 undergraduate students, graduate students, and professionals. The trainees responded positively to the anchored instruction. This article details the module's development, evaluation, and…

  10. Using Anchored Instruction to Evaluate Mathematical Growth and Understanding

    Science.gov (United States)

    Kurz, Terri L.; Batarelo, Ivana

    2005-01-01

    Anchored instruction is designed to present problems in a meaningful context to allow for investigations into real life environments. The Jasper Project was created to allow students to investigate mathematical dilemmas using anchored instruction techniques. This study uses case study methods to examine the perceptions that preservice teachers…

  11. Electrically insulated MLI and thermal anchor

    Science.gov (United States)

    Kamiya, Koji; Furukawa, Masato; Hatakenaka, Ryuta; Miyakita, Takeshi; Murakami, Haruyuki; Kizu, Kaname; Tsuchiya, Katsuhiko; Koidea, Yoshihiko; Yoshida, Kiyoshi

    2014-01-01

    The thermal shield of JT-60SA is kept at 80 K and will use the multilayer insulation (MLI) to reduce radiation heat load to the superconducting coils at 4.4 K from the cryostat at 300 K. Due to plasma pulse operation, the MLI is affected by eddy current in toroidal direction. The MLI is designed to suppress the current by electrically insulating every 20 degree in the toroidal direction by covering the MLI with polyimide films. In this paper, two kinds of designs for the MLI system are proposed, focusing on a way to overlap the layers. A boil-off calorimeter method and temperature measurement has been performed to determine the thermal performance of the MLI system. The design of the electrical insulated thermal anchor between the toroidal field (TF) coil and the thermal shield is also explained.

  12. Structure of a CGI-58 Motif Provides the Molecular Basis of Lipid Droplet Anchoring*

    Science.gov (United States)

    Boeszoermenyi, Andras; Nagy, Harald Manuel; Arthanari, Haribabu; Pillip, Christoph Jens; Lindermuth, Hanna; Luna, Rafael Eulogio; Wagner, Gerhard; Zechner, Rudolf; Zangger, Klaus; Oberer, Monika

    2015-01-01

    Triacylglycerols (TGs) stored in lipid droplets (LDs) are hydrolyzed in a highly regulated metabolic process called lipolysis to free fatty acids that serve as energy substrates for β-oxidation, precursors for membrane lipids and signaling molecules. Comparative gene identification-58 (CGI-58) stimulates the enzymatic activity of adipose triglyceride lipase (ATGL), which catalyzes the hydrolysis of TGs to diacylglycerols and free fatty acids. In adipose tissue, protein-protein interactions between CGI-58 and the LD coating protein perilipin 1 restrain the ability of CGI-58 to activate ATGL under basal conditions. Phosphorylation of perilipin 1 disrupts these interactions and mobilizes CGI-58 for the activation of ATGL. We have previously demonstrated that the removal of a peptide at the N terminus (residues 10–31) of CGI-58 abrogates CGI-58 localization to LDs and CGI-58-mediated activation of ATGL. Here, we show that this tryptophan-rich N-terminal peptide serves as an independent LD anchor, with its three tryptophans serving as focal points of the left (harboring Trp21 and Trp25) and right (harboring Trp29) anchor arms. The solution state NMR structure of a peptide comprising the LD anchor bound to dodecylphosphocholine micelles as LD mimic reveals that the left arm forms a concise hydrophobic core comprising tryptophans Trp21 and Trp25 and two adjacent leucines. Trp29 serves as the core of a functionally independent anchor arm. Consequently, simultaneous tryptophan alanine permutations in both arms abolish localization and activity of CGI-58 as opposed to tryptophan substitutions that occur in only one arm. PMID:26350461

  13. Expanding the clinical and molecular characteristics of PIGT-CDG, a disorder of glycosylphosphatidylinositol anchors.

    Science.gov (United States)

    Lam, Christina; Golas, Gretchen A; Davids, Mariska; Huizing, Marjan; Kane, Megan S; Krasnewich, Donna M; Malicdan, May Christine V; Adams, David R; Markello, Thomas C; Zein, Wadih M; Gropman, Andrea L; Lodish, Maya B; Stratakis, Constantine A; Maric, Irina; Rosenzweig, Sergio D; Baker, Eva H; Ferreira, Carlos R; Danylchuk, Noelle R; Kahler, Stephen; Garnica, Adolfo D; Bradley Schaefer, G; Boerkoel, Cornelius F; Gahl, William A; Wolfe, Lynne A

    2015-01-01

    PIGT-CDG, an autosomal recessive syndromic intellectual disability disorder of glycosylphosphatidylinositol (GPI) anchors, was recently described in two independent kindreds [Multiple Congenital Anomalies-Hypotonia-Seizures Syndrome 3 (OMIM, #615398)]. PIGT encodes phosphatidylinositol-glycan biosynthesis class T, a subunit of the heteropentameric transamidase complex that facilitates the transfer of GPI to proteins. GPI facilitates attachment (anchoring) of proteins to cell membranes. We describe, at ages 7 and 6 years, two children of non-consanguineous parents; they had hypotonia, severe global developmental delay, and intractable seizures along with endocrine, ophthalmologic, skeletal, hearing, and cardiac anomalies. Exome sequencing revealed that both siblings had compound heterozygous variants in PIGT (NM_015937.5), i.e., c.918dupC, a novel duplication leading to a frameshift, and c.1342C > T encoding a previously described missense variant. Flow cytometry studies showed decreased surface expression of GPI-anchored proteins on granulocytes, consistent with findings in previous cases. These siblings further delineate the clinical spectrum of PIGT-CDG, reemphasize the neuro-ophthalmologic presentation, clarify the endocrine features, and add hypermobility, low CSF albumin quotient, and hearing loss to the phenotypic spectrum. Our results emphasize that GPI anchor-related congenital disorders of glycosylation (CDGs) should be considered in subjects with early onset severe seizure disorders and dysmorphic facial features, even in the presence of a normal carbohydrate-deficient transferrin pattern and N-glycan profiling. Currently available screening for CDGs will not reliably detect this family of disorders, and our case reaffirms that the use of flow cytometry and genetic testing is essential for diagnosis in this group of disorders.

  14. Scaffold State Switching Amplifies, Accelerates, and Insulates Protein Kinase C Signaling*

    Science.gov (United States)

    Greenwald, Eric C.; Redden, John M.; Dodge-Kafka, Kimberly L.; Saucerman, Jeffrey J.

    2014-01-01

    Scaffold proteins localize two or more signaling enzymes in close proximity to their downstream effectors. A-kinase-anchoring proteins (AKAPs) are a canonical family of scaffold proteins known to bind protein kinase A (PKA) and other enzymes. Several AKAPs have been shown to accelerate, amplify, and specify signal transduction to dynamically regulate numerous cellular processes. However, there is little theory available to mechanistically explain how signaling on protein scaffolds differs from solution biochemistry. In our present study, we propose a novel kinetic mechanism for enzymatic reactions on protein scaffolds to explain these phenomena, wherein the enzyme-substrate-scaffold complex undergoes stochastic state switching to reach an active state. This model predicted anchored enzymatic reactions to be accelerated, amplified, and insulated from inhibition compared with those occurring in solution. We exploited a direct interaction between protein kinase C (PKC) and AKAP7α as a model to validate these predictions experimentally. Using a genetically encoded PKC activity reporter, we found that both the strength and speed of substrate phosphorylation were enhanced by AKAP7α. PKC tethered to AKAP7α was less susceptible to inhibition from the ATP-competitive inhibitor Gö6976 and the substrate-competitive inhibitor PKC 20-28, but not the activation-competitive inhibitor calphostin C. Model predictions and experimental validation demonstrated that insulation is a general property of scaffold tethering. Sensitivity analysis indicated that these findings may be applicable to many other scaffolds as well. Collectively, our findings provide theoretical and experimental evidence that scaffold proteins can amplify, accelerate, and insulate signal transduction. PMID:24302730

  15. Breaking anchored droplets in a microfluidic Hele-Shaw cell

    CERN Document Server

    Amselem, Gabriel; Gallaire, François; Baroud, Charles N

    2015-01-01

    We study microfluidic self digitization in Hele-Shaw cells using pancake droplets anchored to surface tension traps. We show that above a critical flow rate, large anchored droplets break up to form two daughter droplets, one of which remains in the anchor. Below the critical flow velocity for breakup the shape of the anchored drop is given by an elastica equation that depends on the capillary number of the outer fluid. As the velocity crosses the critical value, the equation stops admitting a solution that satisfies the boundary conditions; the drop breaks up in spite of the neck still having finite width. A similar breaking event also takes place between the holes of an array of anchors, which we use to produce a 2D array of stationary drops in situ.

  16. Effects of accuracy motivation and anchoring on metacomprehension judgment and accuracy.

    Science.gov (United States)

    Zhao, Qin

    2012-01-01

    The current research investigates how accuracy motivation impacts anchoring and adjustment in metacomprehension judgment and how accuracy motivation and anchoring affect metacomprehension accuracy. Participants were randomly assigned to one of six conditions produced by the between-subjects factorial design involving accuracy motivation (incentive or no) and peer performance anchor (95%, 55%, or no). Two studies showed that accuracy motivation did not impact anchoring bias, but the adjustment-from-anchor process occurred. Accuracy incentive increased anchor-judgment gap for the 95% anchor but not for the 55% anchor, which induced less certainty about the direction of adjustment. The findings offer support to the integrative theory of anchoring. Additionally, the two studies revealed a "power struggle" between accuracy motivation and anchoring in influencing metacomprehension accuracy. Accuracy motivation could improve metacomprehension accuracy in spite of anchoring effect, but if anchoring effect is too strong, it could overpower the motivation effect. The implications of the findings were discussed.

  17. Grapnel stone anchors from Saurashtra: Remnants of Indo-Arab trade on the Indian coast

    Digital Repository Service at National Institute of Oceanography (India)

    Gaur, A.S.; Sundaresh; Tripati, S.

    Stone anchors have been used as a primary source of information on ancient navigation by marine archaeologists since long. These anchors used by ancient mariners are often noticed underwater at various places across the world. Stone anchors are also...

  18. A novel membrane anchor for FtsZ is linked to cell wall hydrolysis in Caulobacter crescentus.

    Science.gov (United States)

    Meier, Elizabeth L; Razavi, Shiva; Inoue, Takanari; Goley, Erin D

    2016-07-01

    In most bacteria, the tubulin-like GTPase FtsZ forms an annulus at midcell (the Z-ring) which recruits the division machinery and regulates cell wall remodeling. Although both activities require membrane attachment of FtsZ, few membrane anchors have been characterized. FtsA is considered to be the primary membrane tether for FtsZ in bacteria, however in Caulobacter crescentus, FtsA arrives at midcell after stable Z-ring assembly and early FtsZ-directed cell wall synthesis. We hypothesized that additional proteins tether FtsZ to the membrane and demonstrate that in C. crescentus, FzlC is one such membrane anchor. FzlC associates with membranes directly in vivo and in vitro and recruits FtsZ to membranes in vitro. As for most known membrane anchors, the C-terminal peptide of FtsZ is required for its recruitment to membranes by FzlC in vitro and midcell recruitment of FzlC in cells. In vivo, overproduction of FzlC causes cytokinesis defects whereas deletion of fzlC causes synthetic defects with dipM, ftsE and amiC mutants, implicating FzlC in cell wall hydrolysis. Our characterization of FzlC as a novel membrane anchor for FtsZ expands our understanding of FtsZ regulators and establishes a role for membrane-anchored FtsZ in the regulation of cell wall hydrolysis.

  19. Distributed localization for anchor-free sensor networks

    Institute of Scientific and Technical Information of China (English)

    Cui Xunxue; Shan Zhiguan; Liu Jianjun

    2008-01-01

    Geographic location of nodes is very useful in a sensor network. Previous localization algorithms assume that there exist some anchor nodes in this kind of network, and then other nodes are estimated to create their coordinates. Once there are not anchors to be deployed, those localization algorithms will be invalidated. Many papers in this field focus on anchor-based solutions. The use of anchors introduces many limitations, since anchors require external equipments such as global position system, cause additional power consumption. A novel positioning algorithm is proposed to use a virtual coordinate system based on a new concept-virtual anchor. It is executed in a distributed fashion according to the connectivity of a node and the measured distances to its neighbors. Both the adjacent member information and the ranging distance result are combined to generate the estimated position of a network, one of which is independently adopted for localization previously. At the position refinement stage the intermediate estimation of a node begins to be evaluated on its reliability for position mutation; thus the positioning optimization process of the whole network is avoided falling into a local optimal solution. Simulation results prove that the algorithm can resolve the distributed localization problem for anchor-free sensor networks, and is superior to previous methods in terms of its positioning capability under a variety of circumstances.

  20. Anchors for effectors: subversion of phosphoinositide lipids by Legionella

    Directory of Open Access Journals (Sweden)

    Hubert eHilbi

    2011-04-01

    Full Text Available The facultative intracellular pathogen Legionella pneumophila replicates in free-living amoebae and macrophages within a distinct compartment, the Legionella-containing vacuole (LCV. LCV formation involves phosphoinositide (PI glycerolipids, which are key factors controlling vesicle trafficking pathways and membrane dynamics of eukaryotic cells. To govern the interactions with host cells, L. pneumophila employs the Icm/Dot type IV secretion system and more than 250 translocated effector proteins that presumably subvert host signaling and vesicle trafficking pathways. Some of the effector proteins anchor through distinct PIs to the cytosolic face of LCVs and promote the interaction with host vesicles and organelles, catalyze guanine nucleotide exchange of small GTPases, or bind to PI-metabolizing enzymes, such as OCRL1. The PI 5-phosphatase OCRL1 and its Dictyostelium homologue Dd5P4 restrict intracellular growth of L. pneumophila. Moreover, OCRL1/Dd5P4, PI 3-kinases (PI3Ks and PI4KIIIβ regulate LCV formation and localization of the effector protein SidC, which selectively decorates the LCV membrane. SidC or its 20 kDa P4C fragment are robust and specific probes for phosphatidylinositol-4-phosphate, and SidC can be targeted to purify intact LCVs by immuno-magnetic separation. Taken together, bacterial PI-binding effectors as well as host PIs and PI-modulating enzymes play a pivotal role for intracellular replication of L. pneumophila, and the PI-binding effectors are valuable tools for the analysis of eukaryotic PI lipids.

  1. Neurostimulation leads, intrathecal catheters and anchoring devices evolution

    Directory of Open Access Journals (Sweden)

    Laura Demartini

    2015-03-01

    Full Text Available Many scientific studies highlight the usefulness of spinal cord stimulation and intrathecal therapy for the management of chronic pain syndromes resistant to pharmacological or less invasive interventional therapies. One of the possible complications of these techniques, reported in literature, is migration of the lead or catheter; thus the use of an anchoring system is considered mandatory. Every company that produces devices for neurostimulation or neuromodulation provides various anchoring devices evolved over time. In the study, the authors discuss about the most common anchoring devices based on their clinical experience.

  2. Multiple sequence alignment with user-defined anchor points

    Directory of Open Access Journals (Sweden)

    Pöhler Dirk

    2006-04-01

    Full Text Available Abstract Background Automated software tools for multiple alignment often fail to produce biologically meaningful results. In such situations, expert knowledge can help to improve the quality of alignments. Results Herein, we describe a semi-automatic version of the alignment program DIALIGN that can take pre-defined constraints into account. It is possible for the user to specify parts of the sequences that are assumed to be homologous and should therefore be aligned to each other. Our software program can use these sites as anchor points by creating a multiple alignment respecting these constraints. This way, our alignment method can produce alignments that are biologically more meaningful than alignments produced by fully automated procedures. As a demonstration of how our method works, we apply our approach to genomic sequences around the Hox gene cluster and to a set of DNA-binding proteins. As a by-product, we obtain insights about the performance of the greedy algorithm that our program uses for multiple alignment and about the underlying objective function. This information will be useful for the further development of DIALIGN. The described alignment approach has been integrated into the TRACKER software system.

  3. Anchored boundary conditions for locally isostatic networks

    Science.gov (United States)

    Theran, Louis; Nixon, Anthony; Ross, Elissa; Sadjadi, Mahdi; Servatius, Brigitte; Thorpe, M. F.

    2015-11-01

    Finite pieces of locally isostatic networks have a large number of floppy modes because of missing constraints at the surface. Here we show that by imposing suitable boundary conditions at the surface the network can be rendered effectively isostatic. We refer to these as anchored boundary conditions. An important example is formed by a two-dimensional network of corner sharing triangles, which is the focus of this paper. Another way of rendering such networks isostatic is by adding an external wire along which all unpinned vertices can slide (sliding boundary conditions). This approach also allows for the incorporation of boundaries associated with internal holes and complex sample geometries, which are illustrated with examples. The recent synthesis of bilayers of vitreous silica has provided impetus for this work. Experimental results from the imaging of finite pieces at the atomic level need such boundary conditions, if the observed structure is to be computer refined so that the interior atoms have the perception of being in an infinite isostatic environment.

  4. Anchoring of development workings in a zone of influence of mining in case of the level anchoring system

    Science.gov (United States)

    Demin, V. F.; Fofanov, O. B.; Demina, T. V.; Yavorskiy, V. V.

    2017-02-01

    Regularities of the change of the stress-strain state of coal containing rock masses, depending on mining-geological factors, were revealed. These factors allow establishing rational parameters of anchoring of wall rocks to enhance the stability of development workings. Specific conditions of the deflected mode, displays of rock pressure, terms of maintenance depending on technological parameters are investigated. Researches allowed determining the degree of their development influence on the efficiency of application of the anchoring of the hollow making and will allow a reasonable application of anchoring certificates, provide stability of the rocks mining and reduce expenses on its realization and maintenance.

  5. Effect of a preload force on anchor system frequency

    Institute of Scientific and Technical Information of China (English)

    Lu Aihong; Xu Jinhai; Liu Haishun

    2013-01-01

    The interrelationship between preload forces and natural frequencies of anchors was obtained from the structure of an anchor and its mechanical characteristics.We established a numerical model for the dynamic analysis of a bolt support system taking into consideration the working surroundings of the anchor.The natural frequency distribution of the system under various preload forces of the anchor was analyzed with ANSYS.Our results show that each order of the system frequency varied with an increase in preload forces.A single order frequency decreased with an increase in the preload force.A preload force affected low-order frequencies more than high-order frequencies.We obtained a functional relationship by fitting preload forces and fundamental frequencies,which was in agreement with our theoretical considerations.This study provides theoretical support for the detection of preload forces.

  6. Reaching the top: career anchors and professional development in nursing.

    Science.gov (United States)

    Kaplan, Ruth; Shmulevitz, Carmela; Raviv, Dennie

    2009-01-01

    This study, based on Shein's conceptual theory of career anchors, examined the relationship between career anchors, professional development and emerging career patterns for graduates of 12 consecutive two year second career programs in nursing (N=231) compared to graduates of concurrent four year academic programs (N=273). A 2-group comparison design was used and data collection tools included a demographic profile, a professional profile and a career anchor questionnaire. Statistically significant differences were found in regard to career anchors (pdevelopment (pnurses were specialization and lifestyle where academic graduates chose management, autonomy and service. Academics displayed a statistically significant preference for administrative specialization (34%) compared to the second career tract (6.5%). Researchers propose that each group develops differently and contributes to the workplace and the importance of both certification and academic incentives to ensure recruitment.

  7. Anchor-induced chondral damage in the hip

    OpenAIRE

    Matsuda, Dean K.; Bharam, Srino; White, Brian J.; Matsuda, Nicole A.; SAFRAN, Marc

    2015-01-01

    The purpose of this study is to investigate the outcomes from anchor-induced chondral damage of the hip, both with and without frank chondral penetration. A multicenter retrospective case series was performed of patients with chondral deformation or penetration during initial hip arthroscopic surgery. Intra-operative findings, post-surgical clinical courses, hip outcome scores and descriptions of arthroscopic treatment in cases requiring revision surgery and anchor removal are reported. Five ...

  8. Medial rectus muscle anchoring in complete oculomotor nerve palsy.

    Science.gov (United States)

    Lee, Si Hyung; Chang, Jee Ho

    2015-10-01

    The management of exotropia resulting from complete oculomotor nerve palsy is challenging. Conventional therapeutic interventions, including supramaximal resection and recession, superior oblique tendon resection and transposition, and several ocular anchoring procedures have yielded less-than-adequate results. Here we describe a novel surgical technique of anchoring the medial rectus muscle to the medial orbital wall in combination with lateral rectus disinsertion and reattachment to the lateral orbital wall.

  9. Behavioral Biases of Individual Investors: The Effect of Anchoring

    Directory of Open Access Journals (Sweden)

    Salma Zaiane

    2015-03-01

    Full Text Available The objective of this paper is to investigate the presence of the anchoring bias in the financial decision making of individual investors. A survey study is carried out to find out how the studied bias affects the investment behavior on the Tunisian stock market. The survey is for exploratory purpose and it is based on multiple factorial correspondence analyses. The results reveal that Tunisian investors do not suffer from the anchoring bias.

  10. Aurora A kinase modulates actin cytoskeleton through phosphorylation of Cofilin: Implication in the mitotic process.

    Science.gov (United States)

    Ritchey, Lisa; Chakrabarti, Ratna

    2014-11-01

    Aurora A kinase regulates early mitotic events through phosphorylation and activation of a variety of proteins. Specifically, Aur-A is involved in centrosomal separation and formation of mitotic spindles in early prophase. The effect of Aur-A on mitotic spindles is mediated by the modulation of microtubule dynamics and association with microtubule binding proteins. In this study we show that Aur-A exerts its effects on spindle organization through the regulation of the actin cytoskeleton. Aurora A phosphorylates Cofilin at multiple sites including S(3) resulting in the inactivation of its actin depolymerizing function. Aur-A interacts with Cofilin in early mitotic phases and regulates its phosphorylation status. Cofilin phosphorylation follows a dynamic pattern during the progression of prophase to metaphase. Inhibition of Aur-A activity induced a delay in the progression of prophase to metaphase. Aur-A inhibitor also disturbed the pattern of Cofilin phosphorylation, which correlated with the mitotic delay. Our results establish a novel function of Aur-A in the regulation of actin cytoskeleton reorganization, through Cofilin phosphorylation during early mitotic stages.

  11. Poor anchoring limits dyslexics' perceptual, memory, and reading skills.

    Science.gov (United States)

    Oganian, Yulia; Ahissar, Merav

    2012-07-01

    The basic deficits underlying the severe and persistent reading difficulties in dyslexia are still highly debated. One of the major topics of debate is whether these deficits are language specific, or affect both verbal and non-verbal stimuli. Recently, Ahissar and colleagues proposed the "anchoring-deficit hypothesis" (Ahissar, Lubin, Putter-Katz, & Banai, 2006), which suggests that dyslexics have a general difficulty in automatic extraction of stimulus regularities from auditory inputs. This hypothesis explained a broad range of dyslexics' verbal and non-verbal difficulties. However, it was not directly tested in the context of reading and verbal memory, which poses the main stumbling blocks to dyslexics. Here we assessed the abilities of adult dyslexics to efficiently benefit from ("anchor to") regularities embedded in repeated tones, orally presented syllables, and written words. We also compared dyslexics' performance to that of individuals with attention disorder (ADHD), but no reading disability. We found an anchoring effect in all groups: all gained from stimulus repetition. However, in line with the anchoring-deficit hypothesis, controls and ADHD participants showed a significantly larger anchoring effect in all tasks. This study is the first that directly shows that the same domain-general deficit, poor anchoring, characterizes dyslexics' performance in perceptual, working memory and reading tasks.

  12. Evaluation of mitral valve replacement anchoring in a phantom

    Science.gov (United States)

    McLeod, A. Jonathan; Moore, John; Lang, Pencilla; Bainbridge, Dan; Campbell, Gordon; Jones, Doug L.; Guiraudon, Gerard M.; Peters, Terry M.

    2012-02-01

    Conventional mitral valve replacement requires a median sternotomy and cardio-pulmonary bypass with aortic crossclamping and is associated with significant mortality and morbidity which could be reduced by performing the procedure off-pump. Replacing the mitral valve in the closed, off-pump, beating heart requires extensive development and validation of surgical and imaging techniques. Image guidance systems and surgical access for off-pump mitral valve replacement have been previously developed, allowing the prosthetic valve to be safely introduced into the left atrium and inserted into the mitral annulus. The major remaining challenge is to design a method of securely anchoring the prosthetic valve inside the beating heart. The development of anchoring techniques has been hampered by the expense and difficulty in conducting large animal studies. In this paper, we demonstrate how prosthetic valve anchoring may be evaluated in a dynamic phantom. The phantom provides a consistent testing environment where pressure measurements and Doppler ultrasound can be used to monitor and assess the valve anchoring procedures, detecting pararvalvular leak when valve anchoring is inadequate. Minimally invasive anchoring techniques may be directly compared to the current gold standard of valves sutured under direct vision, providing a useful tool for the validation of new surgical instruments.

  13. Expression of a kinase-dead form of CPK33 involved in florigen complex formation causes delayed flowering.

    Science.gov (United States)

    Kawamoto, Nozomi; Endo, Motomu; Araki, Takashi

    2015-01-01

    Regulation of flowering time is crucial for reproductive success of plants. FLOWERING LOCUS T (FT) protein is a central component of florigen and forms a ternary complex with 14-3-3 and FD, a basic leucine zipper transcription factor, in the shoot apex and promotes flowering. This complex formation requires phosphorylation of threonine residue at position 282 of FD. A calcium-dependent protein kinase CPK33 is responsible for the phosphorylation. However, possibly due to functional redundancy among calcium-dependent protein kinases, impact of the loss of CPK33 reported in the previous study was rather limited. Here, we report that expression of a kinase-dead form of CPK33 caused a clear delayed-flowering phenotype, supporting for an important role of CPK33 in florigen function through FD phosphorylation.

  14. Pyramidal anchor stone from Baga waters of Goa, west coast of India

    Digital Repository Service at National Institute of Oceanography (India)

    Tripati, S.

    . Pyramidal anchor stones have an apex hole which goes up to the round hole, however Goa anchor stone has no such perforation, but, instead has a rectangular cutting on the apex. The anchor stone is compared with Greek pyramidal anchor stones, and probably...

  15. The nucleoplasmin homolog NLP mediates centromere clustering and anchoring to the nucleolus.

    Science.gov (United States)

    Padeken, Jan; Mendiburo, María José; Chlamydas, Sarantis; Schwarz, Hans-Jürgen; Kremmer, Elisabeth; Heun, Patrick

    2013-04-25

    Centromere clustering during interphase is a phenomenon known to occur in many different organisms and cell types, yet neither the factors involved nor their physiological relevance is well understood. Using Drosophila tissue culture cells and flies, we identified a network of proteins, including the nucleoplasmin-like protein (NLP), the insulator protein CTCF, and the nucleolus protein Modulo, to be essential for the positioning of centromeres. Artificial targeting further demonstrated that NLP and CTCF are sufficient for clustering, while Modulo serves as the anchor to the nucleolus. Centromere clustering was found to depend on centric chromatin rather than specific DNA sequences. Moreover, unclustering of centromeres results in the spatial destabilization of pericentric heterochromatin organization, leading to partial defects in the silencing of repetitive elements, defects during chromosome segregation, and genome instability.

  16. Protein

    Science.gov (United States)

    ... Food Service Resources Additional Resources About FAQ Contact Protein Protein is found throughout the body—in muscle, ... the heart and respiratory system, and death. All Protein Isn’t Alike Protein is built from building ...

  17. Display of GPI-anchored anti-EGFR nanobodies on extracellular vesicles promotes tumour cell targeting

    Directory of Open Access Journals (Sweden)

    Sander A. A. Kooijmans

    2016-03-01

    Full Text Available Background: Extracellular vesicles (EVs are attractive candidate drug delivery systems due to their ability to functionally transport biological cargo to recipient cells. However, the apparent lack of target cell specificity of exogenously administered EVs limits their therapeutic applicability. In this study, we propose a novel method to equip EVs with targeting properties, in order to improve their interaction with tumour cells. Methods: EV producing cells were transfected with vectors encoding for anti-epidermal growth factor receptor (EGFR nanobodies, which served as targeting ligands for tumour cells, fused to glycosylphosphatidylinositol (GPI anchor signal peptides derived from decay-accelerating factor (DAF. EVs were isolated using ultrafiltration/size-exclusion liquid chromatography and characterized using western blotting, Nanoparticle Tracking Analysis, and electron microscopy. EV–tumour cell interactions were analyzed under static conditions using flow cytometry and under flow conditions using a live-cell fluorescence microscopy-coupled perfusion system. Results: V analysis showed that GPI-linked nanobodies were successfully displayed on EV surfaces and were highly enriched in EVs compared with parent cells. Display of GPI-linked nanobodies on EVs did not alter general EV characteristics (i.e. morphology, size distribution and protein marker expression, but greatly improved EV binding to tumour cells dependent on EGFR density under static conditions. Moreover, nanobody-displaying EVs showed a significantly improved cell association to EGFR-expressing tumour cells under flow conditions. Conclusions: We show that nanobodies can be anchored on the surface of EVs via GPI, which alters their cell targeting behaviour. Furthermore, this study highlights GPI-anchoring as a new tool in the EV toolbox, which may be applied for EV display of a variety of proteins, such as antibodies, reporter proteins and signaling molecules.

  18. Display of GPI-anchored anti-EGFR nanobodies on extracellular vesicles promotes tumour cell targeting

    Science.gov (United States)

    Kooijmans, Sander A. A.; Aleza, Clara Gómez; Roffler, Steve R.; van Solinge, Wouter W.; Vader, Pieter; Schiffelers, Raymond M.

    2016-01-01

    Background Extracellular vesicles (EVs) are attractive candidate drug delivery systems due to their ability to functionally transport biological cargo to recipient cells. However, the apparent lack of target cell specificity of exogenously administered EVs limits their therapeutic applicability. In this study, we propose a novel method to equip EVs with targeting properties, in order to improve their interaction with tumour cells. Methods EV producing cells were transfected with vectors encoding for anti-epidermal growth factor receptor (EGFR) nanobodies, which served as targeting ligands for tumour cells, fused to glycosylphosphatidylinositol (GPI) anchor signal peptides derived from decay-accelerating factor (DAF). EVs were isolated using ultrafiltration/size-exclusion liquid chromatography and characterized using western blotting, Nanoparticle Tracking Analysis, and electron microscopy. EV–tumour cell interactions were analyzed under static conditions using flow cytometry and under flow conditions using a live-cell fluorescence microscopy-coupled perfusion system. Results EV analysis showed that GPI-linked nanobodies were successfully displayed on EV surfaces and were highly enriched in EVs compared with parent cells. Display of GPI-linked nanobodies on EVs did not alter general EV characteristics (i.e. morphology, size distribution and protein marker expression), but greatly improved EV binding to tumour cells dependent on EGFR density under static conditions. Moreover, nanobody-displaying EVs showed a significantly improved cell association to EGFR-expressing tumour cells under flow conditions. Conclusions We show that nanobodies can be anchored on the surface of EVs via GPI, which alters their cell targeting behaviour. Furthermore, this study highlights GPI-anchoring as a new tool in the EV toolbox, which may be applied for EV display of a variety of proteins, such as antibodies, reporter proteins and signaling molecules. PMID:26979463

  19. Tripolin A, a novel small-molecule inhibitor of aurora A kinase, reveals new regulation of HURP's distribution on microtubules.

    Directory of Open Access Journals (Sweden)

    Iliana A Kesisova

    Full Text Available Mitotic regulators exhibiting gain of function in tumor cells are considered useful cancer therapeutic targets for the development of small-molecule inhibitors. The human Aurora kinases are a family of such targets. In this study, from a panel of 105 potential small-molecule inhibitors, two compounds Tripolin A and Tripolin B, inhibited Aurora A kinase activity in vitro. In human cells however, only Tripolin A acted as an Aurora A inhibitor. We combined in vitro, in vivo single cell and in silico studies to demonstrate the biological action of Tripolin A, a non-ATP competitive inhibitor. Tripolin A reduced the localization of pAurora A on spindle microtubules (MTs, affected centrosome integrity, spindle formation and length, as well as MT dynamics in interphase, consistent with Aurora A inhibition by RNAi or other specific inhibitors, such as MLN8054 or MLN8237. Interestingly, Tripolin A affected the gradient distribution towards the chromosomes, but not the MT binding of HURP (Hepatoma Up-Regulated Protein, a MT-associated protein (MAP and substrate of the Aurora A kinase. Therefore Tripolin A reveals a new way of regulating mitotic MT stabilizers through Aurora A phosphorylation. Tripolin A is predicted to bind Aurora A similarly but not identical to MLN8054, therefore it could be used to dissect pathways orchestrated by Aurora kinases as well as a scaffold for further inhibitor development.

  20. Tripolin A, a novel small-molecule inhibitor of aurora A kinase, reveals new regulation of HURP's distribution on microtubules.

    Science.gov (United States)

    Kesisova, Iliana A; Nakos, Konstantinos C; Tsolou, Avgi; Angelis, Dimitrios; Lewis, Joe; Chatzaki, Aikaterini; Agianian, Bogos; Giannis, Athanassios; Koffa, Maria D

    2013-01-01

    Mitotic regulators exhibiting gain of function in tumor cells are considered useful cancer therapeutic targets for the development of small-molecule inhibitors. The human Aurora kinases are a family of such targets. In this study, from a panel of 105 potential small-molecule inhibitors, two compounds Tripolin A and Tripolin B, inhibited Aurora A kinase activity in vitro. In human cells however, only Tripolin A acted as an Aurora A inhibitor. We combined in vitro, in vivo single cell and in silico studies to demonstrate the biological action of Tripolin A, a non-ATP competitive inhibitor. Tripolin A reduced the localization of pAurora A on spindle microtubules (MTs), affected centrosome integrity, spindle formation and length, as well as MT dynamics in interphase, consistent with Aurora A inhibition by RNAi or other specific inhibitors, such as MLN8054 or MLN8237. Interestingly, Tripolin A affected the gradient distribution towards the chromosomes, but not the MT binding of HURP (Hepatoma Up-Regulated Protein), a MT-associated protein (MAP) and substrate of the Aurora A kinase. Therefore Tripolin A reveals a new way of regulating mitotic MT stabilizers through Aurora A phosphorylation. Tripolin A is predicted to bind Aurora A similarly but not identical to MLN8054, therefore it could be used to dissect pathways orchestrated by Aurora kinases as well as a scaffold for further inhibitor development.

  1. Perceptual anchoring in preschool children: not adultlike, but there.

    Directory of Open Access Journals (Sweden)

    Karen Banai

    Full Text Available BACKGROUND: Recent studies suggest that human auditory perception follows a prolonged developmental trajectory, sometimes continuing well into adolescence. Whereas both sensory and cognitive accounts have been proposed, the development of the ability to base current perceptual decisions on prior information, an ability that strongly benefits adult perception, has not been directly explored. Here we ask whether the auditory frequency discrimination of preschool children also improves when given the opportunity to use previously presented standard stimuli as perceptual anchors, and whether the magnitude of this anchoring effect undergoes developmental changes. METHODOLOGY/PRINCIPAL FINDINGS: Frequency discrimination was tested using two adaptive same/different protocols. In one protocol (with-reference, a repeated 1-kHz standard tone was presented repeatedly across trials. In the other (no-reference, no such repetitions occurred. Verbal memory and early reading skills were also evaluated to determine if the pattern of correlations between frequency discrimination, memory and literacy is similar to that previously reported in older children and adults. Preschool children were significantly more sensitive in the with-reference than in the no-reference condition, but the magnitude of this anchoring effect was smaller than that observed in adults. The pattern of correlations among discrimination thresholds, memory and literacy replicated previous reports in older children. CONCLUSIONS/SIGNIFICANCE: The processes allowing the use of context to form perceptual anchors are already functional among preschool children, albeit to a lesser extent than in adults. Nevertheless, immature anchoring cannot fully account for the poorer frequency discrimination abilities of young children. That anchoring is present among the majority of typically developing preschool children suggests that the anchoring deficits observed among individuals with dyslexia represent a

  2. Surface analysis of PEGylated nano-shields on nanoparticles installed by hydrophobic anchors

    DEFF Research Database (Denmark)

    Ebbesen, M F; Whitehead, Bradley Joseph; Gonzalez, Borja Ballarin;

    2013-01-01

    and cellular interactions. Methods: Poly(lactic-co-glycolic acid) (PLGA) nanoparticles were prepared with a hydrophilic PEGylated "nano-shield" inserted at different levels by hydrophobic anchoring using either a phospholipid-PEG conjugate or the copolymer PLGA-block-PEG by an emulsification/diffusion method......Purpose: This work describes a method for functionalisation of nanoparticle surfaces with hydrophilic "nano-shields" and the application of advanced surface characterisation to determine PEG amount and accumulation at the outmost 10 nm surface that is the predominant factor in determining protein...

  3. Membrane anchoring stabilizes and favors secretion of New Delhi metallo-β-lactamase.

    Science.gov (United States)

    González, Lisandro J; Bahr, Guillermo; Nakashige, Toshiki G; Nolan, Elizabeth M; Bonomo, Robert A; Vila, Alejandro J

    2016-07-01

    Carbapenems, 'last-resort' β-lactam antibiotics, are inactivated by zinc-dependent metallo-β-lactamases (MBLs). The host innate immune response withholds nutrient metal ions from microbial pathogens by releasing metal-chelating proteins such as calprotectin. We show that metal sequestration is detrimental for the accumulation of MBLs in the bacterial periplasm, because those enzymes are readily degraded in their nonmetallated form. However, the New Delhi metallo-β-lactamase (NDM-1) can persist under conditions of metal depletion. NDM-1 is a lipidated protein that anchors to the outer membrane of Gram-negative bacteria. Membrane anchoring contributes to the unusual stability of NDM-1 and favors secretion of this enzyme in outer-membrane vesicles (OMVs). OMVs containing NDM-1 can protect nearby populations of bacteria from otherwise lethal antibiotic levels, and OMVs from clinical pathogens expressing NDM-1 can carry this MBL and the blaNDM gene. We show that protein export into OMVs can be targeted, providing possibilities of new antibacterial therapeutic strategies.

  4. An Analysis for Cross Beam-Ground Anchor Reinforcement

    Institute of Scientific and Technical Information of China (English)

    Xu Yingzi; Zhang Baiqing; Tang Huiming

    2005-01-01

    With the rapid development of water facilities, hydroelectric projects, highways and railways in China, beam-anchor reinforcement has been widely used to stabilize slopes in recent years. But the theory for the design of beam-anchor reinforcement is far behind the application. Cross beam-ground anchor reinforcement is a combination of beams and anchors forming a new structure to prevent slope sliding. The forces in the beams are discussed using theoretical analysis and numerical modeling. The Winkler model is used to analyze the beams, and reasonable values of λ, length, spacing and cantilevered length for the beams are determined through a theoretical analysis. A three-dimensional finite element method is adopted to model the interaction of the beams and soils and a structure analysis is applied to treat the beams and to study the stress distribution in external and internal beams. The analytical results show that it is better to satisfy λ≥2π, the spacing between anchors ls should be lsλ<π/2 and cantilever length should be (0.3-0.5)ls for the optimum design. The numerical results show that the same design can be used for all beams in different directions, including the internal and external beams. The application of the analytical method for reinforcement beam analysis is acceptable. It is better to choose a safety coefficient of 1.3 for design based on the analytical method for safety.

  5. Structural Requirements for Membrane Assembly of Proteins Spanning the Membrane Several Times

    OpenAIRE

    Lipp, Joachim; Flint, Nicholas; Haeuptle, Marie-Theres; Dobberstein, Bernhard

    1989-01-01

    We have investigated the structural requirements for the biogenesis of proteins spanning the membrane several times. Proteins containing various combinations of topological signals (signal anchor and stop transfer sequences) were synthesized in a cell-free translation system and their membrane topology was determined. Proteins spanning the membrane twice were obtained when a signal anchor sequence was followed by either a stop transfer sequence or a second signal anchor sequence. Thus, a sig...

  6. Optical Fiber Grating Sensor for Force Measurement of Anchor Cable

    Institute of Scientific and Technical Information of China (English)

    JIANG Desheng; FU Jinghua; LIU Shengchun; SUI Lingfeng; FU Rong

    2006-01-01

    The development of the sensor suitable for measuring large load stress to the anchor cable becomes an important task in bridge construction and maintenance. Therefore, a new type of optical fiber sensor was developed in the laboratory - optical fiber grating sensor for force measurement of anchor cable (OFBFMAC). No similar report about this kind of sensor has been found up to now in China and other countries. This sensor is proved to be an effective way of monitoring in processes of anchor cable installation, cable cutting, cable force regulation, etc, with the accurate and repeatable measuring results. Its successful application in the tie bar cable force safety monitoring for Wuhan Qingchuan bridge is a new exploration of optical fiber grating sensing technology in bridge tie bar monitoring system.

  7. Sortase A substrate specificity in GBS pilus 2a cell wall anchoring.

    Science.gov (United States)

    Necchi, Francesca; Nardi-Dei, Vincenzo; Biagini, Massimiliano; Assfalg, Michael; Nuccitelli, Annalisa; Cozzi, Roberta; Norais, Nathalie; Telford, John L; Rinaudo, C Daniela; Grandi, Guido; Maione, Domenico

    2011-01-01

    Streptococcus agalactiae, also referred to as Group B Streptococcus (GBS), is one of the most common causes of life-threatening bacterial infections in infants. In recent years cell surface pili have been identified in several Gram-positive bacteria, including GBS, as important virulence factors and promising vaccine candidates. In GBS, three structurally distinct types of pili have been discovered (pilus 1, 2a and 2b), whose structural subunits are assembled in high-molecular weight polymers by specific class C sortases. In addition, the highly conserved housekeeping sortase A (SrtA), whose main role is to link surface proteins to bacterial cell wall peptidoglycan by a transpeptidation reaction, is also involved in pili cell wall anchoring in many bacteria. Through in vivo mutagenesis, we demonstrate that the LPXTG sorting signal of the minor ancillary protein (AP2) is essential for pilus 2a anchoring. We successfully produced a highly purified recombinant SrtA (SrtA(ΔN40)) able to specifically hydrolyze the sorting signal of pilus 2a minor ancillary protein (AP2-2a) and catalyze in vitro the transpeptidation reaction between peptidoglycan analogues and the LPXTG motif, using both synthetic fluorescent peptides and recombinant proteins. By contrast, SrtA(ΔN40) does not catalyze the transpeptidation reaction with substrate-peptides mimicking sorting signals of the other pilus 2a subunits (the backbone protein and the major ancillary protein). Thus, our results add further insight into the proposed model of GBS pilus 2a assembly, in which SrtA is required for pili cell wall covalent attachment, acting exclusively on the minor accessory pilin, representing the terminal subunit located at the base of the pilus.

  8. Sortase A substrate specificity in GBS pilus 2a cell wall anchoring.

    Directory of Open Access Journals (Sweden)

    Francesca Necchi

    Full Text Available Streptococcus agalactiae, also referred to as Group B Streptococcus (GBS, is one of the most common causes of life-threatening bacterial infections in infants. In recent years cell surface pili have been identified in several Gram-positive bacteria, including GBS, as important virulence factors and promising vaccine candidates. In GBS, three structurally distinct types of pili have been discovered (pilus 1, 2a and 2b, whose structural subunits are assembled in high-molecular weight polymers by specific class C sortases. In addition, the highly conserved housekeeping sortase A (SrtA, whose main role is to link surface proteins to bacterial cell wall peptidoglycan by a transpeptidation reaction, is also involved in pili cell wall anchoring in many bacteria. Through in vivo mutagenesis, we demonstrate that the LPXTG sorting signal of the minor ancillary protein (AP2 is essential for pilus 2a anchoring. We successfully produced a highly purified recombinant SrtA (SrtA(ΔN40 able to specifically hydrolyze the sorting signal of pilus 2a minor ancillary protein (AP2-2a and catalyze in vitro the transpeptidation reaction between peptidoglycan analogues and the LPXTG motif, using both synthetic fluorescent peptides and recombinant proteins. By contrast, SrtA(ΔN40 does not catalyze the transpeptidation reaction with substrate-peptides mimicking sorting signals of the other pilus 2a subunits (the backbone protein and the major ancillary protein. Thus, our results add further insight into the proposed model of GBS pilus 2a assembly, in which SrtA is required for pili cell wall covalent attachment, acting exclusively on the minor accessory pilin, representing the terminal subunit located at the base of the pilus.

  9. Distribution of the novel cell wall anchored protein-encoding gene sasX in Staphylococcus aureus strains%新的细胞壁锚定蛋白编码基因sasX在金黄色葡萄球菌中的分布

    Institute of Scientific and Technical Information of China (English)

    杜昕; 宋燕; 田月如; 阮斐怡; 吕元; 李敏

    2011-01-01

    Objective To investigate the distribution and effect on antibiotic resistance of the novel cell wall anchored protein-encoding gene sasX.Methods A total of 300 S.aureus isolates were randomly collected from inpatients with S.aureus infection in Shanghai Huashan hospital in 2004, 2007 and 2010.Meanwhile,170 S.aureus isolates from the nasal swabs of healthy people were collected as part of a population-based community prevalence study.Typing of S.aureus isolates were identified by using multilocus sequence typing (MLST) and S.aureus-specific staphylococcal protein A typing (spa typing).Determination of oxacillin MICs was used to screen MRSA.PCR and sequencing were used to analyze sasX gene.The effect on antibiotic resistance of sasX gene was detect by disc agar diffusion drug sensitive test.Results The major clonal types of the 300 S.aureus isolates collected from inpatients with S.aureus infection were ST239 ( 110,36.7%) and ST5 (122,40.7%).From 2004 to 2010,the percentage of isolates from inpatients with S.aureus infection was increased from 17% to 39%,but sasX was only found in 0.59% of the S.aureus isolates from the nasal swabs of healthy people.The percentage of sasX positive was increased from 47.2% to 83.8% in ST239.The percentage of sasX positive MRSA was increased from 26.4% to 50.8%,but the percentage of sasX positive MSSA was about 10%.Antibiotic resistance of sasX positive strains were higher than that of sasX negative strains.Conclusions SasX gene is mainly detected in nosocomial pathogenic S.aureus and it is a possible virulence factor of S.aureus in hosptal setting.The presence of sasX gene is related to antibiotic resistance.For better understanding the real function of this novel gene,further studies such as expression of the encoded protein should be carried out.%目的 研究细胞壁锚定蛋白编码基因sasX在金黄色葡萄球菌中的分布及对细菌耐药性的影响.方法 随机收集上海华山医院2004、2007

  10. Experience-based, body-anchored qualitative research interviewing

    DEFF Research Database (Denmark)

    Stelter, Reinhard

    2010-01-01

    Two theoretical constructs that lay the foundation for experience-based, body-anchored interviewing are presented: the first-person perspective and the concept of meaning. These theoretical concepts are concretized, first, by means of a methodological framework for experience-based, body......-anchored interviewing, and second, by an interview guide that explores a research participant's personal experience with mindfulness meditation. An excerpt from an interview is discussed to illustrate the advantages of this interview form, namely its value as a methodological instrument for qualitative research...... in areas such as traditional and holistic medicine, Western alternative and complementary medicine, nursing, psychotherapy, coaching, physiotherapy, movement arts, and physical education....

  11. Mechanical behavior of the U-anchor of super-CFRP rod under tensile loading

    OpenAIRE

    Djamaluddin, Rudy; yamaguchi, Kohei; Hino, Shinichi

    2014-01-01

    - A suitable anchoring system is required to anchor a CFRP tendon due to its sensitivity in lateral pressure. Recent developed anchors are still relying on lateral pressure in anchoring CFRP tendons. A new CFRP unit equipped with U-anchor at both end of the rod body without any jointing (namely of Super CFRP, S-CFRP) has been developed. This paper presents the mechanical behavior as well as failure mechanism of U-anchor under direct loading and loaded under embedded within concrete, respec...

  12. Arrangement of anchor reinforcement in roadway for fully mechanized sublevel caving

    Institute of Scientific and Technical Information of China (English)

    勾攀峰; 陈启永; 芦付松

    2003-01-01

    Bolting of mining roadway for fully mechanized sublevel caving has been practised successfully in Hebi mining area.It provides a new method for roadway support and settles the problem of support difficulty radically for sublevel caving in Hebi mining area.Where anchor reinforcement holds an important station in roadway support.This article brings forward the arrangement project of anchor based on theoretic analysis.Compared with arranged in the middle of the entry, anchor arranged in the vertex of the entry can reduces the length of anchor,shortens the anchor installation time,and heightens the reliability of anchor installation.

  13. AKAP3 synthesis is mediated by RNA binding proteins and PKA signaling during mouse spermiogenesis.

    Science.gov (United States)

    Xu, Kaibiao; Yang, Lele; Zhao, Danyun; Wu, Yaoyao; Qi, Huayu

    2014-06-01

    Mammalian spermatogenesis is regulated by coordinated gene expression in a spatiotemporal manner. The spatiotemporal regulation of major sperm proteins plays important roles during normal development of the male gamete, of which the underlying molecular mechanisms are poorly understood. A-kinase anchoring protein 3 (AKAP3) is one of the major components of the fibrous sheath of the sperm tail that is formed during spermiogenesis. In the present study, we analyzed the expression of sperm-specific Akap3 and the potential regulatory factors of its protein synthesis during mouse spermiogenesis. Results showed that the transcription of Akap3 precedes its protein synthesis by about 2 wk. Nascent AKAP3 was found to form protein complex with PKA and RNA binding proteins (RBPs), including PIWIL1, PABPC1, and NONO, as revealed by coimmunoprecipitation and protein mass spectrometry. RNA electrophoretic gel mobility shift assay showed that these RBPs bind sperm-specific mRNAs, of which proteins are synthesized during the elongating stage of spermiogenesis. Biochemical and cell biological experiments demonstrated that PIWIL1, PABPC1, and NONO interact with each other and colocalize in spermatids' RNA granule, the chromatoid body. In addition, NONO was found in extracytoplasmic granules in round spermatids, whereas PIWIL1 and PABPC1 were diffusely localized in cytoplasm of elongating spermatids, indicating their participation at different steps of mRNA metabolism during spermatogenesis. Interestingly, type I PKA subunits colocalize with PIWIL1 and PABPC1 in the cytoplasm of elongating spermatids and cosediment with the RBPs in polysomal fractions on sucrose gradients. Further biochemical analyses revealed that activation of PKA positively regulates AKAP3 protein synthesis without changing its mRNA level in elongating spermatids. Taken together, these results indicate that PKA signaling directly participates in the regulation of protein translation in postmeiotic male germ cells

  14. Expression of GPI anchored human recombinant erythropoietin in CHO cells is devoid of glycosylation heterogeneity.

    Science.gov (United States)

    Singh, Pankaj Kumar; Devasahayam, Mercy; Devi, Sobita

    2015-04-01

    Erythropoietin is a glycohormone involved in the regulation of the blood cell levels. It is a 166 amino acid protein having 3 N-glycosylation and one O-linked glycosylation sites, and is used to treat anaemia related illness. Though human recombinant erythropoietin (rEPO) is produced in CHO cells, the loss in quality control is 80% due to incomplete glycosylation of the rEPO with low levels of fully glycosylated active rEPO. Here, we describe the expression from CHO cells of fully glycosylated human rEPO when expressed as a GPI anchored molecule (rEPO-g). The results demonstrated the production of a homogenous completely glycosylated human rEPO-g as a 42 kD band without any low molecular weight glycoform variants as shown by affinity chromatography followed by SDS-PAGE and anti-human EPO specific western blot. The western blot using specific monoclonal antibody is the available biochemical technique to prove the presence of homogeneity in the expressed recombinant protein. The GPI anchor can be removed during the purification process to yield a therapeutically relevant recombinant erythropoietin molecule cells with a higher in vivo biological activity due to its high molecular weight of 40 kD. This is possibly the first report on the production of a homogenous and completely glycosylated human rEPO from CHO cells for efficient therapy.

  15. Current status of frameless anchored IUD for immediate intracesarean insertion.

    Science.gov (United States)

    Wildemeersch, Dirk; Goldstuck, Norman D; Hasskamp, Thomas

    2016-01-01

    Immediate postpartum intrauterine device (IUD) insertion deserves great attention as it can provide immediate, timely and convenient contraception plus the added benefit of preventing repeat unintended pregnancies. Although women post vaginal delivery can benefit from immediate post-placenta contraception, women undergoing Cesarean section clearly need contraception, as an inter-delivery interval shorter than 18 months places them at a high risk for uterine rupture. The main drawback of currently available framed IUD devices for immediate postpartum insertion of an IUD is their high expulsion and displacement rates when inserted immediately postpartum after both vaginal and Cesarean delivery. Current research suggests that a brief window of opportunity exists of 10 minutes for insertion of conventional IUDs after which time expulsion rates both immediately and over time are greatly enhanced. This paper summarizes the current research conducted to overcome the expulsion problems associated with conventional T-shaped devices as well as through the use of an anchored frameless device. In the 1970s and 1980s, attempts were made to solve the expulsion problem by modifying existing devices, such as adding absorbable sutures (Delta-T) or additional appendages. These attempts proved to be clinically unsuccessful as the catgut suture added to the transverse arms did not provide sufficient resistance to prevent downward displacement and expulsion. An anchoring technique to suspend a copper IUD to the fundus of the uterus was developed in Belgium in the 1980s and has been the subject of extensive ongoing clinical research since 1985. Recently the frameless copper releasing anchor IUD, GyneFix, has been tested for postplacental insertion. Initially, the anchor was modified by the inclusion of a biodegradable cone which was added below the anchoring knot. Clinical studies confirmed the adequacy of this approach suggesting that it was technically possible to anchor an IUD

  16. Stepping stones of Amsterdam, Local and global anchor points 2010.

    NARCIS (Netherlands)

    Bois, G. de Peter

    2011-01-01

    This report shows the results of research on the relation between local and global anchor points in the city of Amsterdam, executed by a small group of international students from Italy and Spain within the scope of two research modules in their Bachelor education at the University of Applied Scienc

  17. Finding the Optimal Guidance for Enhancing Anchored Instruction

    Science.gov (United States)

    Zydney, Janet Mannheimer; Bathke, Arne; Hasselbring, Ted S.

    2014-01-01

    This study investigated the effect of different methods of guidance with anchored instruction on students' mathematical problem-solving performance. The purpose of this research was to iteratively design a learning environment to find the optimal level of guidance. Two iterations of the software were compared. The first iteration used explicit…

  18. Implementing Anchored Instruction: Guiding Principles for Curriculum Development.

    Science.gov (United States)

    McLarty, Kim; And Others

    A curriculum based on "anchored instruction" was developed to enhance students' literacy development and acquisition of knowledge. The curriculum was designed to create a rich, shared environment that generates interest and enables students to identify and define problems while they explore the content from many perspectives. Based on…

  19. Analytical Model for Hook Anchor Pull-out

    DEFF Research Database (Denmark)

    Brincker, Rune; Ulfkjær, J. P.; Adamsen, P.;

    A simple analytical model for the pull-out of a hook anchor is presented. The model is based on a simplified version of the fictitious crack model. It is assumed that the fracture process is the pull-off of a cone shaped concrete part, simplifying the problem by assuming pure rigid body motions...

  20. Analytical Model for Hook Anchor Pull-Out

    DEFF Research Database (Denmark)

    Brincker, Rune; Ulfkjær, J. P.; Adamsen, P.;

    1995-01-01

    A simple analytical model for the pull-out of a hook anchor is presented. The model is based on a simplified version of the fictitious crack model. It is assume that the fracture process is the pull-off of a cone shaped concrete part, simplifying the problem by assuming pure rigid body motions...

  1. Almost Lie structures on an anchored Banach bundle

    CERN Document Server

    Cabau, Patrick

    2011-01-01

    Under appropriate assumptions, we generalize the concept of linear almost Poisson struc- tures, almost Lie algebroids, almost differentials in the framework of Banach anchored bundles and the relation between these objects. We then obtain an adapted formalism for mechanical systems which is illustrated by the evolutionary problem of the "Hilbert snake"

  2. Modified Anchor Shaped Post Core Design for Primary Anterior Teeth

    OpenAIRE

    R. Rajesh; Kusai Baroudi; K. Bala Kasi Reddy; Praveen, B. H.; V. Sumanth Kumar; Amit, S

    2014-01-01

    Restoring severely damaged primary anterior teeth is challenging to pedodontist. Many materials are tried as a post core but each one of them has its own drawbacks. This a case report describing a technique to restore severely damaged primary anterior teeth with a modified anchor shaped post. This technique is not only simple and inexpensive but also produces better retention.

  3. Modified Anchor Shaped Post Core Design for Primary Anterior Teeth

    Directory of Open Access Journals (Sweden)

    R. Rajesh

    2014-01-01

    Full Text Available Restoring severely damaged primary anterior teeth is challenging to pedodontist. Many materials are tried as a post core but each one of them has its own drawbacks. This a case report describing a technique to restore severely damaged primary anterior teeth with a modified anchor shaped post. This technique is not only simple and inexpensive but also produces better retention.

  4. Modified anchor shaped post core design for primary anterior teeth.

    Science.gov (United States)

    Rajesh, R; Baroudi, Kusai; Reddy, K Bala Kasi; Praveen, B H; Kumar, V Sumanth; Amit, S

    2014-01-01

    Restoring severely damaged primary anterior teeth is challenging to pedodontist. Many materials are tried as a post core but each one of them has its own drawbacks. This a case report describing a technique to restore severely damaged primary anterior teeth with a modified anchor shaped post. This technique is not only simple and inexpensive but also produces better retention.

  5. Ten Anchor Points for Teaching Principles of Marketing

    Science.gov (United States)

    Tomkovick, Chuck

    2004-01-01

    Effective marketing instructors commonly share a love for their students, an affinity for the subject matter, and a devotion to continuous quality improvement. The purpose of this article is to highlight 10 anchor points for teaching Principles of Marketing, which are designed to better engage students in the learning process. These anchor…

  6. Conventional Anchor Test Results at San Diego and Indian Island

    Science.gov (United States)

    1980-07-01

    lines passed through open chocks welded at opposite ends of the barge. A crane barge was used to install and recover all test anchors at each site...DEPT OF FISH & GAME Long Beach CA (Marine Tech Info Ctr) CALIF. DEPT OF NAVIGATION & OCEAN DEV. Sacramento, CA (G. Armstrong) CALIF. MARITIME ACADEMY

  7. The bone-anchored hearing aid for children: recent developments.

    NARCIS (Netherlands)

    Snik, A.F.M.; Leijendeckers, J.M.; Hol, M.K.S.; Mylanus, E.A.M.; Cremers, C.

    2008-01-01

    In 1984 the Bone-Anchored Hearing Aid, or BAHA, system was introduced. Its transducer is coupled directly to the skull percutaneously to form a highly effective bone-conduction hearing device. Clinical studies on adults with conductive hearing loss have shown that the BAHA system outperforms convent

  8. Stone anchors from the Okhamandal region, Gujarat Coast, India

    Digital Repository Service at National Institute of Oceanography (India)

    Sundaresh; Gaur, A.S.; Gudigar, P.; Tripati, S.; Vora, K.H.; Bandodkar, S.N.

    period particularly on south-western coast of the island while stone anchors were noticed on the eastern coast of the island where medieval period remains can be noticed in the form of pottery, cannons and a few stone structures. Similar type of stone...

  9. Anchoring return: the role of the Solutions Strategy

    Directory of Open Access Journals (Sweden)

    Pierfrancesco Maria Natta

    2014-05-01

    Full Text Available Providing a minimum standard of living and livelihood opportunities to help anchor those who have returned is critical for the future stability and security of Afghanistan. This is one of the three main objectives of the 2012 Solutions Strategy for Afghan Refugees.

  10. The importance of anchor text for ad hoc search revisited

    NARCIS (Netherlands)

    M. Koolen; J. Kamps

    2010-01-01

    It is generally believed that propagated anchor text is very important for effective Web search as offered by the commercial search engines. "Google Bombs" are a notable illustration of this. However, many years of TREC Web retrieval research failed to establish the effectiveness of link evidence fo

  11. Impact of Enhanced Anchored Instruction in Inclusive Math Classrooms

    Science.gov (United States)

    Bottge, Brian A.; Toland, Michael D.; Gassaway, Linda; Butler, Mark; Choo, Sam; Griffen, Ann Katherine; Ma, Xin

    2015-01-01

    The Common Core State Standards for Mathematics will place more pressure on special education and math teachers to raise the skill levels of all students, especially those with disabilities in math (MD). The purpose of this study was to assess the effects of enhanced anchored instruction (EAI) on students with and without MD in co-taught general…

  12. Fullerene-based Anchoring Groups for Molecular Electronics

    DEFF Research Database (Denmark)

    Martin, Christian A.; Ding, Dapeng; Sørensen, Jakob Kryger

    2008-01-01

    We present results on a new fullerene-based anchoring group for molecular electronics. Using lithographic mechanically controllable break junctions in vacuum we have determined the conductance and stability of single-molecule junctions of 1,4-bis(fullero[c]pyrrolidin-1-yl)benzene. The compound can...

  13. Poor Anchoring Limits Dyslexics' Perceptual, Memory, and Reading Skills

    Science.gov (United States)

    Oganian, Yulia; Ahissar, Merav

    2012-01-01

    The basic deficits underlying the severe and persistent reading difficulties in dyslexia are still highly debated. One of the major topics of debate is whether these deficits are language specific, or affect both verbal and non-verbal stimuli. Recently, Ahissar and colleagues proposed the "anchoring-deficit hypothesis" (Ahissar, Lubin,…

  14. The Effect of Anchor Test Construction on Scale Drift

    Science.gov (United States)

    Antal, Judit; Proctor, Thomas P.; Melican, Gerald J.

    2014-01-01

    In common-item equating the anchor block is generally built to represent a miniature form of the total test in terms of content and statistical specifications. The statistical properties frequently reflect equal mean and spread of item difficulty. Sinharay and Holland (2007) suggested that the requirement for equal spread of difficulty may be too…

  15. Audiometric evaluation of bilaterally fitted bone-anchored hearing aids.

    NARCIS (Netherlands)

    Bosman, A.J.; Snik, A.F.M.; Pouw, C.T.M. van der; Mylanus, E.A.M.; Cremers, C.W.R.J.

    2001-01-01

    Bilateral fittings of bone-anchored hearing aids (BAHA) were evaluated in 25 patients with at least 3 months experience with using two BAHAs. For all patients, air conduction hearing aids were contraindicated due to either recurrent otorrhoea or otitis externa (19 cases) or to congenital aural atres

  16. The biggest stone anchors (Indo-Arab type) from Mithi Virdi, Bhavnagar, west coast of India

    Digital Repository Service at National Institute of Oceanography (India)

    Gaur, A.S.; Sundaresh; Kumar, B.

    A coastal archaeological exploration in Talaja taluka of Bhavnagar district, Gujarat, brought to light 5 grapnel stone anchors in an agriculture land. The site and these anchors are known as Bhima ka Khatiya ('Cot of Bhima', a famous character...

  17. Synergistic effect of a new wedge-bond-type anchor for CFRP tendons

    Institute of Scientific and Technical Information of China (English)

    谢桂华; 刘荣桂; 陈蓓; 李明君; 石天罡

    2015-01-01

    In order to improve the anchoring force of anchors for carbon fiber reinforced polymer (CFRP) tendons further, a new wedge-bond-type anchor for CFRP tendons was developed. The increment in anchoring force induced by the clamping segment of anchor was studied. Taking the deformation of all parts in clamping segment in the transverse direction into consideration, the calculation formula for the increment of anchoring force was proposed based on the linear elastic hypotheses. The proposed model is verified by experiments and conclusions are drawn that the anchoring force is influenced mainly by the inclination angle of clamping pieces, the length of clamping part and the thickness of bonding medium. Especially, the thickness of bonding medium should be lowered in design to improve the synergistic effect of anchors.

  18. Radiocarbon dates of the medieval period stone anchors from Dabhol, west coast of India

    Digital Repository Service at National Institute of Oceanography (India)

    Gaur, A; Sundaresh; Tripati, S.; Vora, K.H.

    Dabhol was an important medieval-period port town on the Maharashtra coast. An archaeological exploration at Dabhol has revealed four stone anchors with remains of wood in fluke hole. These anchors were retrieved from Dabhol creek during a dredging...

  19. A soluble acid invertase is directed to the vacuole by a signal anchor mechanism.

    Science.gov (United States)

    Rae, Anne L; Casu, Rosanne E; Perroux, Jai M; Jackson, Mark A; Grof, Christopher P L

    2011-06-15

    Enzyme activities in the vacuole have an important impact on the net concentration of sucrose. In sugarcane (Saccharum hybrid), immunolabelling demonstrated that a soluble acid invertase (β-fructofuranosidase; EC 3.2.1.26) is present in the vacuole of storage parenchyma cells during sucrose accumulation. Examination of sequences from sugarcane, barley and rice showed that the N-terminus of the invertase sequence contains a signal anchor and a tyrosine motif, characteristic of single-pass membrane proteins destined for lysosomal compartments. The N-terminal peptide from the barley invertase was shown to be capable of directing the green fluorescent protein to the vacuole in sugarcane cells. The results suggest that soluble acid invertase is sorted to the vacuole in a membrane-bound form.

  20. 新的细胞壁锚定蛋白SasX对金黄葡萄球菌生物膜形成和毒力的影响%The novel surface-anchored protein SasX promotes biofilm formation and is a virulence factor of Staphylococcus aureus

    Institute of Scientific and Technical Information of China (English)

    杜昕; 宋燕; 胡锦辉; 阮斐怡; 吕元; 李敏

    2011-01-01

    Objective To determine whether the novel surface-anchored protein SasX mediates biofilm formation of Staphylococcus aureus and to investigate the function of SasX in the virulence of S.aureus.Methods Methicillin-resistant Staphylococcus aureus(MRSA) ST-239 HS sasX gene mutant (HS770 △ sasX) and complement [ HS770 △sasX(pRBsasX) ] were gotten by gene knock out and complement methods.Semiquantitative biofilm assay was used for detection of the biofilm formation of wild type and mutant.By using abscess model in mice,we investigate the function of SasX in the virulence of S.aureus.Results The sasX gene mutant strain was gotten successfully by using pKOR1 plasmid.HS770 △sasX with a very clear reduction of biofilm formation compared to wild type and complement(P<0.05),there was no significant difference of biofilm formation between wild type and complement(P>0.05).Primary attachment assays demonstrated that comparing to wild type,there was significant reduction of initial accumulative phases of biofilm development in HS770△sasX(P<0.05),but there was no difference between wild type and complement(P>0.05).Both sasX wild type and mutant could cause abscess in the skin of mice,no abscess were forming in the PBS control group.The biggest abscess size was found on the second day after injection.The size of abscess was smaller with extension of the time.But on the same time,the wild type group produced significantly larger abscesses compared to mutant group( P<0.05 ).Conclusion It could improve the effect of gene knocking out by using pKOR1 plasmid.SasX promotes biofilm formation by influencing on the initial accumulative phases of biofilm development,SasX is a marker of invasive infection of S.aureus,it is very important for S.aureus persistence in the hospital setting.%目的 研究新的细胞壁锚定蛋白SasX对金黄葡萄球菌生物膜形成及毒力的影响.方法 采用基因敲除及基因互补技术获得耐甲

  1. Biomimetic Mussel Adhesive Inspired Clickable Anchors Applied to the Functionalization of Fe3O4 Nanoparticles

    NARCIS (Netherlands)

    Goldmann, Anja S.; Schoedel, Christine; Walther, Andreas; Yuan, Jiayin; Loos, Katja; Mueller, Axel H. E.; Müller, Axel H.E.

    2010-01-01

    The functionalization of magnetite (Fe3O4) nanoparticles with dopamine-derived clickable biomimetic anchors is reported. Herein, an alkyne-modified catechol-derivative is employed as the anchor, as i) the catechol-functional anchor groups possess irreversible covalent binding affinity to Fe3O4 nanop

  2. Perception of the Raison d'Etre in Anchored Instruction: An Ecological Psychology Perspective.

    Science.gov (United States)

    Young, Michael F.; Barab, Sasha A.

    1999-01-01

    Anchored instruction calls for the establishment of a macrocontext to "anchor" instruction within a realistic situation. Evidence is provided that video anchors encourage students to adopt certain contrived goals over their more naturalistic goals. Suggests that goals that enable the problem solver to detect the "raison d'etre"…

  3. The influence of bone formation on anchoring percutaneous devices with titanium fibre mesh flanges.

    NARCIS (Netherlands)

    Shalabi, M.M.; Walboomers, X.F.; Jansen, J.A.

    2004-01-01

    For man-made percutaneous devices (PD), it is known that anchoring will improve the clinical success. Previously, our Department has designed PDs that use a sheet of titanium (Ti) fibre mesh for anchoring. In nature, a very successful natural PD occurs, namely the tooth. Teeth are anchored in the al

  4. A group of 20 stone anchors from the waters of Dwarka, on the Gujarat Coast, India

    Digital Repository Service at National Institute of Oceanography (India)

    Gaur, A.S; Sundaresh; Tripati, S; Gudigar, P.; Vora, K.H.; Bandodkar, S

    A large number of stone anchors were discovered in a water depth of 10-14 m off Dwarka during the 1998-99 season. The seabed near the anchors consists of a ledge with an average height of 1 m. Several anchors were found trapped between the rocks...

  5. Improvement in organophosphorus hydrolase activity of cell surface-engineered yeast strain using Flo1p anchor system

    OpenAIRE

    Fukuda, Takeshi; Tsuchiyama, Kouta; Makishima, Hirokazu; Takayama, Katsumi; Mulchandani, Ashok; Kuroda, Kouichi; Ueda, Mitsuyoshi; Suye, Shin-ichiro

    2010-01-01

    Organophosphorus hydrolase (OPH) hydrolyzes organophosphorus esters. We constructed the yeast-displayed OPH using Flo1p anchor system. In this system, the N-terminal region of the protein was fused to Flo1p and the fusion protein was displayed on the cell surface. Hydrolytic reactions with paraoxon were carried out during 24 h of incubation of OPH-displaying cells at 30°C. p-Nitrophenol produced in the reaction mixture was detected by HPLC. The strain with highest activity showed 8-fold great...

  6. The Golgin GMAP210/TRIP11 anchors IFT20 to the Golgi complex.

    Directory of Open Access Journals (Sweden)

    John A Follit

    2008-12-01

    Full Text Available Eukaryotic cells often use proteins localized to the ciliary membrane to monitor the extracellular environment. The mechanism by which proteins are sorted, specifically to this subdomain of the plasma membrane, is almost completely unknown. Previously, we showed that the IFT20 subunit of the intraflagellar transport particle is localized to the Golgi complex, in addition to the cilium and centrosome, and hypothesized that the Golgi pool of IFT20 plays a role in sorting proteins to the ciliary membrane. Here, we show that IFT20 is anchored to the Golgi complex by the golgin protein GMAP210/Trip11. Mice lacking GMAP210 die at birth with a pleiotropic phenotype that includes growth restriction, ventricular septal defects of the heart, omphalocele, and lung hypoplasia. Cells lacking GMAP210 have normal Golgi structure, but IFT20 is no longer localized to this organelle. GMAP210 is not absolutely required for ciliary assembly, but cilia on GMAP210 mutant cells are shorter than normal and have reduced amounts of the membrane protein polycystin-2 localized to them. This work suggests that GMAP210 and IFT20 function together at the Golgi in the sorting or transport of proteins destined for the ciliary membrane.

  7. A myristoyl/phosphoserine switch controls cAMP-dependent protein kinase association to membranes.

    Science.gov (United States)

    Gaffarogullari, Ece C; Masterson, Larry R; Metcalfe, Emily E; Traaseth, Nathaniel J; Balatri, Erica; Musa, Musa M; Mullen, Daniel; Distefano, Mark D; Veglia, Gianluigi

    2011-08-26

    The cAMP-dependent protein kinase [protein kinase A (PKA)] mediates a myriad of cellular signaling events, and its activity is tightly regulated in both space and time. Among these regulatory mechanisms is N-myristoylation, whose biological role has been elusive. Using a combination of thermodynamics, kinetics, and spectroscopic methods, we analyzed the effects of N-myristoylation and phosphorylation at Ser10 on the interactions of PKA with model membranes. We found that, in the absence of lipids, the myristoyl group is tucked into the hydrophobic binding pocket of the enzyme (myr-in state). Upon association with lipid bilayers, the myristoyl group is extruded and inserts into the hydrocarbon region of the lipid bilayer (myr-out state). NMR data indicate that the enzyme undergoes conformational equilibrium between myr-in and myr-out states, which can be shifted byeither interaction with membranes and/or phosphorylation at Ser10. Our results provide evidence that the membrane binding motif of the myristoylated C-subunit of PKA (PKA-C) steers the enzyme toward lipids independent of its regulatory subunit or an A-kinase anchoring protein, providing an additional mechanism to localize the enzyme near membrane-bound substrates.

  8. NCBI nr-aa BLAST: CBRC-MDOM-02-0386 [SEVENS

    Lifescience Database Archive (English)

    Full Text Available CBRC-MDOM-02-0386 sp|Q02952|AKA12_HUMAN RecName: Full=A-kinase anchor protein 12; AltName: Full...=A-kinase anchor protein 250 kDa; Short=AKAP 250; AltName: Full=Gravin; AltName: Full=Myasthenia gravis autoantigen Q02952 0.0 35% ...

  9. Hydrophobic profiles of the tail anchors in SLMAP dictate subcellular targeting

    Directory of Open Access Journals (Sweden)

    Salih Maysoon

    2009-06-01

    Full Text Available Abstract Background Tail anchored (TA membrane proteins target subcellular structures via a C-terminal transmembrane domain and serve prominent roles in membrane fusion and vesicle transport. Sarcolemmal Membrane Associated Protein (SLMAP possesses two alternatively spliced tail anchors (TA1 or TA2 but their specificity of subcellular targeting remains unknown. Results TA1 or TA2 can direct SLMAP to reticular structures including the endoplasmic reticulum (ER, whilst TA2 directs SLMAP additionally to the mitochondria. Despite the general structural similarity of SLMAP to other vesicle trafficking proteins, we found no evidence for its localization with the vesicle transport machinery or a role in vesicle transport. The predicted transmembrane region of TA2 is flanked on either side by a positively charged amino acid and is itself less hydrophobic than the transmembrane helix present in TA1. Substitution of the positively charged amino acids, in the regions flanking the transmembrane helix of TA2, with leucine did not alter its subcellular targeting. The targeting of SLMAP to the mitochondria was dependent on the hydrophobic nature of TA2 since targeting of SLMAP-TA2 was prevented by the substitution of leucine (L for moderately hydrophobic amino acid residues within the transmembrane region. The SLMAP-TA2-4L mutant had a hydrophobic profile that was comparable to that of SLMAP-TA1 and had identical targeting properties to SLMAP-TA1. Conclusion Thus the overall hydrophobicity of the two alternatively spliced TAs in SLMAP determines its subcellular targeting and TA2 predominantly directs SLMAP to the mitochondira where it may serve roles in the function of this organelle.

  10. Rapid Deterioration of the Anchor Performance of the Wind Turbine Caused by Liquid Water

    OpenAIRE

    千々和, 伸浩; Chijiwa, Nobuhiro; MAI, THI HONG; Mai.Thi, Hong; 岩波, 光保; Iwanami, Mitsuyasu

    2015-01-01

    In some wind turbines, it is found that sludge spouting on the tower foundation and the tower is lifted up just after several months have passed from starting service. Common characteristic of those wind turbine towers is that they are anchored to the foundation by anchor ring method. In anchor ring method, ring-shaped base plate welded to the wind turbine tower is expected to act as anchor in the foundation. It is suspected that liquid water may have some effect on such a kind of the anchor ...

  11. Research on roll stabilization for ships at anchor

    Institute of Scientific and Technical Information of China (English)

    JIN Hong-zhang; QI Zhi-gang; ZHOU Ting; Li Dong-song

    2008-01-01

    With the increasing importance of ocean exploitation,providing anti-rolling stability for ships at anchor has become more and more important. The lift-generation theory of traditional fin stabilizers is based on incoming flow velocity,which is not suitable for explaining lift generated at anchor. We analyzed non-steady flows,with forces on fin stabilizers generated by non-incoming flow velocity conditions,and gave a new lift-generation model. The correctness of the model was proven by comparing experimental results of fin stabilizer motion under non-incoming velocity conditions from the fluid computation software with that from the emulator of the lift-generation model. Finally,the model was used in an anti-rolling system on a ship and the reduction of roll was much better than what could be achieved by passive anti-rolling tanks.

  12. Graphene nanoribbons anchored to SiC substrates

    Science.gov (United States)

    Le, Nam B.; Woods, Lilia M.

    2016-09-01

    Graphene nanoribbons are quasi-one-dimensional planar graphene allotropes with diverse properties dependent on their width and types of edges. Graphene nanoribbons anchored to substrates is a hybrid system, which offers novel opportunities for property modifications as well as experimental control. Here we present electronic structure calculations of zigzag graphene nanoribbons chemically attached via the edges to the Si or C terminated surfaces of a SiC substrate. The results show that the edge characteristics are rather robust and the properties are essentially determined by the individual nanoribbon. While the localized spin polarization of the graphene nanoribbon edge atoms is not significantly affected by the substrate, secondary energy gaps in the highest conduction and lowest valence region may emerge in the anchored structures. The van der Waals interaction together with the electrostatic interactions due to the polarity of the surface bonds are found to be important for the structure parameters and energy stability.

  13. Modification of the Bankart reconstruction using a suture anchor.

    Science.gov (United States)

    Traina, S M; Holtgrewe, J L; King, S

    1998-01-01

    A prospective study was done to determine the effectiveness of a suture anchor in doing a Modified Bankart Reconstruction on the traumatic unidirectional Bankart lesion shoulder. From 1989 to 1991, 26 patients encompassing 27 shoulders with recurrent instability had modified Bankart reconstructions. A minimum 18-month follow-up was obtained by examination of 24 patients with telephone interviews done on two patients. The average follow-up was 23.6 months. A 93.1% good-to-excellent result was obtained using the Bankart rating scale. There were no failures or complications. The surgeon involved in the study thought the suture anchor facilitated the attachment of the avulsed capsulolabral complex in doing the Bankart procedure.

  14. Optimization Analysis Model of Self-Anchored Suspension Bridge

    Directory of Open Access Journals (Sweden)

    Pengzhen Lu

    2014-01-01

    Full Text Available The hangers of self-anchored suspension bridge need to be tensioned suitably during construction. In view of this point, a simplified optimization calculation method of cable force for self-anchored suspension bridge has been developed based on optimization theories, such as minimum bending energy method, and internal force balanced method, influence matrix method. Meanwhile, combined with the weak coherence of main cable and the adjacently interaction of hanger forces, a simplified analysis method is developed using MATLAB, which is then compared with the optimization method that consider the main cable's geometric nonlinearity with software ANSYS in an actual example bridge calculation. This contrast proves the weak coherence of main cable displacement and the limitation of the adjacent cable force influence. Furthermore, a tension program that is of great reference value has been developed; some important conclusions, advices, and attention points have been summarized.

  15. Mobile Anchor Assisted Node Localization for Wireless Sensor Networks

    CERN Document Server

    Chen, Hongyang; Huang, Pei; Poor, H Vincent; Sezaki, Kaoru

    2009-01-01

    In this paper, a cooperative localization algorithm is proposed that considers the existence of obstacles in mobilityassisted wireless sensor networks (WSNs). In this scheme, a mobile anchor (MA) node cooperates with static sensor nodes and moves actively to refine location performance. The localization accuracy of the proposed algorithm can be improved further by changing the transmission range of mobile anchor node. The algorithm takes advantage of cooperation betweenMAs and static sensors while, at the same time, taking into account the relay node availability to make the best use of beacon signals. For achieving high localization accuracy and coverage, a novel convex position estimation algorithm is proposed, which can effectively solve the localization problem when infeasible points occur because of the effects of radio irregularity and obstacles. This method is the only range-free based convex method to solve the localization problem when the feasible set of localization inequalities is empty. Simulatio...

  16. Anchoring FRP laminates for the seismic strengthening of RC columns

    OpenAIRE

    Sadone, Raphaëlle; Quiertant, Marc; FERRIER, Emmanuel; Chataigner, Sylvain; Mercier, Julien

    2012-01-01

    This paper aims to examine the effectiveness of seismic strengthening of reinforced concrete (RC) columns by externally bonded Fibre Reinforced Polymer (FRP). Particularly, a novel strengthening system, designed for the flexural strengthening of columns is studied. This flexural strengthening is achieved by FRP plates bonded longitudinally and anchored at the column-stub junction. The proposed system is validated through an experimental campaign carried out on full-scale RC columns. Different...

  17. Effect of anchoring 4-anilidopiperidines to opioid peptides

    Science.gov (United States)

    Petrov, Ravil R.; Lee, Yeon Sun; Vardanyan, Ruben S.; Liu, Lu; Ma, Shou-wu; Davis, Peg; Lai, Josephine; Porreca, Frank; Vanderah, Todd W.; Hruby, Victor J.

    2014-01-01

    We report here the design, synthesis, and in vitro characterization of new opioid peptides featuring a 4-anilidopiperidine moiety. Despite the fact that the chemical structures of fentanyl surrogates have been found suboptimal per se for the opioid activity, the corresponding conjugates with opioid peptides displayed potent opioid activity. These studies shed an instructive light on the strategies and potential therapeutic values of anchoring the 4-anilidopiperidine scaffold to different classes of opioid peptides. PMID:23623418

  18. Robust dithiocarbamate-anchored amine functionalization of Au nanoparticles

    Science.gov (United States)

    Chen, Kai; Robinson, Hans D.

    2011-02-01

    We introduce an effective and facile technique that achieves robust amine functionalization of Au nanoparticles by binding the polyamine poly(allylamine hydrochloride) (PAH) to the surface using a dithiocarbamate (DTC) modification of the side group amines. The DTC anchor confers superior short- and long-term colloidal stability compared to a physisorbed layer of the same polymer. We also demonstrate that the surface amines are available for further functionalization and that at least four alternately charged polyelectrolyte layers can be assembled onto the particles. The latter modification could not be performed on a physisorbed functional layer, so this demonstrates the effectiveness of the DTC groups in robustly anchoring the polymer to the particle surface. At the same time, the DTC-anchored polymer layer is less than 2 nm thick in the dry state. This is one-third of the thickness of a physisorbed polyamine layer deposited under the same conditions, and sufficiently thin that the plasmonic field enhancement on the metal particle remains accessible to the outside environment. We attribute the difference in thickness to multiple DTC bonds on each polymer chain forcing it into much closer conformity to the particle surface than in the physisorbed case.

  19. Phosphonate-anchored monolayers for antibody binding to magnetic nanoparticles.

    Science.gov (United States)

    Benbenishty-Shamir, Helly; Gilert, Roni; Gotman, Irena; Gutmanas, Elazar Y; Sukenik, Chaim N

    2011-10-04

    Targeted delivery of magnetic iron oxide nanoparticles (IONPs) to a specific tissue can be achieved by conjugation with particular biological ligands on an appropriately functionalized IONP surface. To take best advantage of the unique magnetic properties of IONPs and to maximize their blood half-life, thin, strongly bonded, functionalized coatings are required. The work reported herein demonstrates the successful application of phosphonate-anchored self-assembled monolayers (SAMs) as ultrathin coatings for such particles. It also describes a new chemical approach to the anchoring of antibodies on the surface of SAM-coated IONPs (using nucleophilic aromatic substitution). This anchoring strategy results in stable, nonhydrolyzable, covalent attachment and allows the reactivity of the particles toward antibody binding to be activated in situ, such that prior to the activation the modified surface is stable for long-term storage. While the SAMs do not have the well-packed crystallinity of other such monolayers, their structure was studied using smooth model substrates based on an iron oxide layer on a double-side polished silicon wafer. In this way, atomic force microscopy, ellipsometry, and contact angle goniometry (tools that could not be applied to the nanoparticles' surfaces) could contribute to the determination of their monomolecular thickness and uniformity. Finally, the successful conjugation of IgG antibodies to the SAM-coated IONPs such that the antibodies retain their biological activity is verified by their complexation to a secondary fluorescent antibody.

  20. Influence of anchoring on miscarriage risk perception associated with amniocentesis.

    Science.gov (United States)

    Nuccio, Regina; Hashmi, S Shahrukh; Mastrobattista, Joan; Noblin, Sarah Jane; Refuerzo, Jerrie; Smith, Janice L; Singletary, Claire N

    2015-04-01

    One factor women consider when deciding whether to pursue amniocentesis is the risk of miscarriage. People use mechanisms like anchoring, or the prior belief regarding the magnitude of risk, as a frame of reference for new information. This study aimed to determine a woman's perception of miscarriage risk associated with amniocentesis before and after genetic counseling and to determine what factors anchor a woman's perception of miscarriage risk. One hundred thirteen women being seen for prenatal genetic counseling and possible amniocentesis at six Houston clinics participated in the two-part anonymous survey. While most women (56.7 %) perceived the risk as low or average pre-counseling and indicated the numeric risk of amniocentesis as amniocentesis had a significantly lower perception of the risk (p = 0.017) whereas those who declined amniocentesis were more likely to view the risk as high (p = 0.004). The only two anchoring factors that had an effect were having a friend or relative with a personal or family history of a genetic disorder (p = 0.001) and having a child already (p = 0.038); both were associated with a lower risk perception. The lack of significant factors may reflect the uniqueness of each patient's risk assessment framework and reinforces the importance of genetic counseling to elucidate individual concerns, particularly as non-invasive prenatal testing becomes more widely available and further complicates the prenatal testing landscape.

  1. N-acetylglucosamine deacetylases modulate the anchoring of the gamma-glutamyl capsule to the cell wall of Bacillus anthracis.

    Science.gov (United States)

    Candela, Thomas; Balomenou, Stavroula; Aucher, Willy; Bouriotis, Vassilis; Simore, Jean-Pierre; Fouet, Agnes; Boneca, Ivo G

    2014-06-01

    Bacillus anthracis has a complex cell wall structure composed of a peptidoglycan (PG) layer to which major structures are anchored such as a neutral polysaccharide, an S-layer, and a poly-γ-D-glutamate (PDGA) capsule. Many of these structures have central roles in the biology of B. anthracis, particularly, in virulence. However, little attention has been devoted to structurally study the PG and how it is modified in the presence of these secondary cell wall components. We present here the fine structure of the PG of the encapsulated RPG1 strain harboring both pXO1 and pXO2 virulence plasmids. We show that B. anthracis has a high degree of cross-linking and its GlcNAc residues are highly modified by N-deacetylation. The PG composition is not dependent on the presence of either LPXTG proteins or the capsule. Using NMR analysis of the PG-PDGA complex, we provide evidence for the anchoring of the PDGA to the glucosamine residues. We show that anchoring of the PDGA capsule is impaired in two PG N-deacetylase mutants, Ba1961 and Ba3679. Thus, these multiple N-deactylase activities would constitute excellent drug targets in B. anthracis by simultaneously affecting its resistance to lysozyme and to phagocytosis impairing B. anthracis survival in the host.

  2. Quantitative evaluation of Candia antarctica lipase B displayed on the cell surface of a Pichia pastoris based on an FS anchor system.

    Science.gov (United States)

    Liang, Xing-xiang; Wang, Bei-bei; Sun, Yu-fei; Lin, Ying; Han, Shuang-yan; Zheng, Sui-ping; Cui, Tang-bing

    2013-03-01

    A new approach is described to quantify the number of enzyme molecules, such as Candia antarctica lipase B, that are displayed on the cell surface of Pichia pastoris. Enhanced green fluorescent protein (EGFP) and Candida antarctica lipase B (CALB) were fused and displayed on the surface of P. pastoris by linking to the anchor flocculation functional domain of FLO1p from Saccharomyces cerevisiae. Confocal laser scanning microscopy, flow cytometry, and fluorescence spectrophotometry were used to monitor the fluorescence intensity of fused EGFP. Combined with the corresponding protein concentration detected in the medium, a standard curve describing the relationship between the fusion protein concentration and fluorescence intensity were obtained and could be used to number CALB displayed on the cell surface. The results showed that approx. 10(4) molecules of CALB molecules were immobilized on the single P. pastoris cell wall based on FS anchor system.

  3. Spatially confined polymer chains: implications of chromatin fibre flexibility and peripheral anchoring on telomere telomere interaction

    Science.gov (United States)

    Gehlen, L. R.; Rosa, A.; Klenin, K.; Langowski, J.; Gasser, S. M.; Bystricky, K.

    2006-04-01

    We simulate the extension of spatially confined chromatin fibres modelled as polymer chains and examine the effect of the flexibility of the fibre and its degree of freedom. The developed formalism was used to analyse experimental data of telomere-telomere distances in living yeast cells in the absence of confining factors as identified by the proteins Sir4 and yKu70. Our analysis indicates that intrinsic properties of the chromatin fibre, in particular its elastic properties and flexibility, can influence the juxtaposition of the telomeric ends of chromosomes. However, measurements in intact yeast cells showed that the telomeres of chromosomes 3 and 6 come even closer together than the parameters of constraint imposed on the simulations would predict. This juxtaposition was specific to telomeres on one contiguous chromosome and overrode a tendency for separation that is imposed by anchoring.

  4. Construction and Expression of Bivalent Membrane-anchored DNA Vaccine Encoding Sj14FABP and Sj26GST Genes

    Institute of Scientific and Technical Information of China (English)

    GUO Ping; DAI Wuxing; LIU Shuojie; YANG Ping; CHENG Jizhong; LIANG Liang; CHEN Zhihao; GAO Hong

    2006-01-01

    In order to construct a eukaryotic co-expression plasmid containing membrane-anchored Sjcl4FABP and Sjc26GST genes and identify their expression in vitro, Sj14 and Sj26 genes were obtained by RT-PCR with total RNA of Schistosoma japonicum adult worms as the template and cloned into eukaryotic expression plasmid pVAC to construct recombinant plasmids pVAC-Sj14 and pVAC-Sj26. Then a 23 amino-acid signal peptide of human interleukin-2 (IL-2) upstream Sj14 or Sj26 gene and a membrane-anchored sequence containing 32 amino-acids of carboxyl-terminal of human placental alkaline phosphatase (PLAP) downstream were amplified by PCR as the template of plasmid pVAC-Sjl4 or pVAC-Sj26 only to get two gene fragments including Sjl4 gene and Sj26 gene. The two modified genes were altogether cloned into a eukaryotic co-expression plasmid pIRES,resulting in another new recombinant plasmid pIRES-Sj26-Sj 14. The expression of Sj14 and Sj26genes was detected by RT-PCR and indirect immunofluorescent assays (IFA) when the plasmid pIRES-Sj26-Sj 14 was transfected into eukaryotic Hela cells. Restriction enzyme analysis, PCR and sequencing results revealed that the recombinant plasmids pVAC-Sj14, pVAC-Sj26 and pIRES-Sj26-Sj14 were successfully constructed and the expression of modified Sj 14 and Sj26 genes could be detected by RT-PCR and IFA. A bivalent membrane-anchored DNA vaccine encoding Sj14 and Sj26 genes was acquired and expressed proteins were proved to be mostly anchored in cellular membranes.

  5. A novel glycosylphosphatidylinositol-anchored glycoside hydrolase from Ustilago esculenta functions in β-1,3-glucan degradation.

    Science.gov (United States)

    Nakajima, Masahiro; Yamashita, Tetsuro; Takahashi, Machiko; Nakano, Yuki; Takeda, Takumi

    2012-08-01

    A glycoside hydrolase responsible for laminarin degradation was partially purified to homogeneity from a Ustilago esculenta culture filtrate by weak-cation-exchange, strong-cation-exchange, and size-exclusion chromatography. Three proteins in enzymatically active fractions were digested with chymotrypsin followed by liquid chromatography-tandem mass spectrometry (LC/MS/MS) analysis, resulting in the identification of three peptide sequences that shared significant similarity to a putative β-1,3-glucanase, a member of glucoside hydrolase family 16 (GH16) from Sporisorium reilianum SRZ2. A gene encoding a laminarin-degrading enzyme from U. esculenta, lam16A, was isolated by PCR using degenerate primers designed based on the S. reilianum SRZ2 β-1,3-glucanase gene. Lam16A possesses a GH16 catalytic domain with an N-terminal signal peptide and a C-terminal glycosylphosphatidylinositol (GPI) anchor peptide. Recombinant Lam16A fused to an N-terminal FLAG peptide (Lam16A-FLAG) overexpressed in Aspergillus oryzae exhibited hydrolytic activity toward β-1,3-glucan specifically and was localized both in the extracellular and in the membrane fractions but not in the cell wall fraction. Lam16A without a GPI anchor signal peptide was secreted extracellularly and was not detected in the membrane fraction. Membrane-anchored Lam16A-FLAG was released completely by treatment with phosphatidylinositol-specific phospholipase C. These results suggest that Lam16A is anchored in the plasma membrane in order to modify β-1,3-glucan associated with the inner cell wall and that Lam16A is also used for the catabolism of β-1,3-glucan after its release in the extracellular medium.

  6. Two-Dimensional Large Deformation Finite Element Analysis for the Pulling-up of Plate Anchor

    Institute of Scientific and Technical Information of China (English)

    WANG Dong; HU Yu-xia; JIN Xia

    2006-01-01

    Based on mesh regeneration and stress interpolation from an old mesh to a new one, a large deformation finite element model is developed for the study of the behaviour of circular plate anchors subjected to uplift loading. For the determination of the distributions of stress components across a clay foundation, the Recovery by Equilibrium in Patches is extended to plastic analyses. ABAQUS, a commercial finite element package, is customized and linked into our program so as to keep automatic and efficient running of large deformation calculation. The quality of stress interpolation is testified by evaluations of Tresca stress and nodal reaction forces. The complete pulling-up processes of plate anchors buried in homogeneous clay are simulated, and typical pulling force-displacement responses of a deep anchor and a shallow anchor are compared. Different from the results of previous studies, large deformation analysis is of the capability of estimating the breakaway between the anchor bottom and soils. For deep anchors, the variation of mobilized uplift resistance with anchor settlement is composed of three stages, and the initial buried depths of anchors affect the separation embedment slightly. The uplift bearing capacity of deep anchors is usually higher than that of shallow anchors.

  7. Not all nutrition claims are perceived equal: anchoring effects and moderating mechanisms in food advertising.

    Science.gov (United States)

    Paek, Hye-Jin; Yoon, Hye Jin; Hove, Thomas

    2011-03-01

    Despite the increased use of health claims in food advertising, few studies have investigated how specific nutrition claims have differential effects depending on how they are presented. In this context, the current study tests the anchoring hypothesis. Anchoring refers to a common human tendency to evaluate information differently depending on the presence or absence of a numerical "anchor" or reference point. Two (pilot and main) experimental studies explore anchoring effects on audience response to food advertising both directly and moderated by cognitive, motivational, and message factors. The pilot study finds that food product ads employing nutrition claims with an anchor rather than without an anchor generate two results: First, participants perceive the product to have lower fat/lower calorie contents (anchoring hypothesis); second, they prefer the messages with an anchor over those without an anchor. The main study reports that when anchoring is successfully evoked, it produces favorable attitudes toward the ad, favorable attitudes toward the brand, and purchase intention-but only when moderated by health orientation, claim believability, and nutrition knowledge. Practical implications are provided with respect to regulatory guidelines and effective communication strategies for promoting low-fat and low-calorie products in food advertising.

  8. Relevant uses of surface proteins – display on self‐organized biological structures

    OpenAIRE

    Jahns, Anika C.; Rehm, Bernd H. A.

    2012-01-01

    Summary Proteins are often found attached to surfaces of self‐assembling biological units such as whole microbial cells or subcellular structures, e.g. intracellular inclusions. In the last two decades surface proteins were identified that could serve as anchors for the display of foreign protein functions. Extensive protein engineering based on structure–function data enabled efficient display of technically and/or medically relevant protein functions. Small size, diversity of the anchor pro...

  9. A GPI-anchored alkaline phosphatase is a functional midgut receptor of Cry11Aa toxin in Aedes aegypti larvae.

    Science.gov (United States)

    Fernandez, Luisa E; Aimanova, Karlygash G; Gill, Sarjeet S; Bravo, Alejandra; Soberón, Mario

    2006-02-15

    A 65 kDa GPI (glycosylphosphatidyl-inositol)-anchored ALP (alkaline phosphatase) was characterized as a functional receptor of the Bacillus thuringiensis subsp. israelensis Cry11Aa toxin in Aedes aegypti midgut cells. Two (a 100 kDa and a 65 kDa) GPI-anchored proteins that bound Cry11Aa toxin were preferentially extracted after treatment of BBMV (brush boder membrane vesicles) from Ae. aegypti midgut epithelia with phospholipase C. The 65 kDa protein was further purified by toxin affinity chromatography. The 65 kDa protein showed ALP activity. The peptide-displaying phages (P1.BBMV and P8.BBMV) that bound to the 65 kDa GPI-ALP (GPI-anchored ALP) and competed with the Cry11Aa toxin to bind to BBMV were isolated by selecting BBMV-binding peptide-phages by biopanning. GPI-ALP was shown to be preferentially distributed in Ae. aegypti in the posterior part of the midgut and in the caeca, by using P1.BBMV binding to fixed midgut tissue sections to determine the location of GPI-ALP. Cry11Aa binds to the same regions of the midgut and competed with P1.BBMV and P8.BBMV to bind to BBMV. The importance of this interaction was demonstrated by the in vivo attenuation of Cry11Aa toxicity in the presence of these phages. Our results shows that GPI-ALP is an important receptor molecule involved in Cry11Aa interaction with midgut cells and toxicity to Ae. aegypti larvae.

  10. The cystic fibrosis transmembrane recruiter the alter ego of CFTR as a multi-kinase anchor.

    Science.gov (United States)

    Mehta, Anil

    2007-11-01

    This review focuses on a newly discovered interaction between protein kinases involved in cellular energetics, a process that may be disturbed in cystic fibrosis for unknown reasons. I propose a new model where kinase-mediated cellular transmission of energy provides mechanistic insight to a latent role of the cystic fibrosis transmembrane conductance regulator (CFTR). I suggest that CFTR acts as a multi-kinase recruiter to the apical epithelial membrane. My group finds that, in the cytosol, two protein kinases involved in cell energy homeostasis, nucleoside diphosphate kinase (NDPK) and AMP-activated kinase (AMPK), bind one another. Preliminary data suggest that both can also bind CFTR (function unclear). The disrupted role of this CFTR-kinase complex as 'membrane transmitter to the cell' is proposed as an alternative paradigm to the conventional ion transport mediated and CFTR/chloride-centric view of cystic fibrosis pathogenesis. Chloride remains important, but instead, chloride-induced control of the phosphohistidine content of one kinase component (NDPK, via a multi-kinase complex that also includes a third kinase, CK2; formerly casein kinase 2). I suggest that this complex provides the necessary near-equilibrium conditions needed for efficient transmission of phosphate energy to proteins controlling cellular energetics. Crucially, a new role for CFTR as a kinase controller is proposed with ionic concentration acting as a signal. The model posits a regulatory control relay for energy sensing involving a cascade of protein kinases bound to CFTR.

  11. Clustering and Functional Coupling of Diverse Ion Channels and Signaling Proteins Revealed by Super-resolution STORM Microscopy in Neurons.

    Science.gov (United States)

    Zhang, Jie; Carver, Chase M; Choveau, Frank S; Shapiro, Mark S

    2016-10-19

    The fidelity of neuronal signaling requires organization of signaling molecules into macromolecular complexes, whose components are in intimate proximity. The intrinsic diffraction limit of light makes visualization of individual signaling complexes using visible light extremely difficult. However, using super-resolution stochastic optical reconstruction microscopy (STORM), we observed intimate association of individual molecules within signaling complexes containing ion channels (M-type K(+), L-type Ca(2+), or TRPV1 channels) and G protein-coupled receptors coupled by the scaffolding protein A-kinase-anchoring protein (AKAP)79/150. Some channels assembled as multi-channel supercomplexes. Surprisingly, we identified novel layers of interplay within macromolecular complexes containing diverse channel types at the single-complex level in sensory neurons, dependent on AKAP79/150. Electrophysiological studies revealed that such ion channels are functionally coupled as well. Our findings illustrate the novel role of AKAP79/150 as a molecular coupler of different channels that conveys crosstalk between channel activities within single microdomains in tuning the physiological response of neurons.

  12. Clinical Outcomes After Suture Anchor Repair of Recalcitrant Medial Epicondylitis.

    Science.gov (United States)

    Grawe, Brian M; Fabricant, Peter D; Chin, Christopher S; Allen, Answorth A; DePalma, Brian J; Dines, David M; Altchek, David W; Dines, Joshua S

    2016-01-01

    This study evaluated clinical and patient-reported outcomes and return to sport after surgical treatment of medial epicondylitis with suture anchor fixation. Consecutive patients were evaluated after undergoing debridement and suture anchor repair of the flexor-pronator mass for the treatment of medial epicondylitis. Demographic variables, a short version of the Disabilities of the Arm, Shoulder and Hand (QuickDASH) score, Oxford Elbow Score (OES), and 10-point pain and satisfaction scales were collected. Ability and time to return to sport after surgery were evaluated, and the relationship between predictor variables and both elbow function and return to sport was investigated. Median age at the time of surgery was 55 years (range, 29-65 years), with median follow-up of 40 months (range, 12-67 months). Median QuickDASH score and OES at final follow-up were 2.3 (range, 0-38.6) and 45 (range, 22-48), respectively. Most patients returned to premorbid sporting activities at a median of 4.5 months (range, 2.5-12 months), whereas 4 patients (14%) reported significant limitations at final follow-up. Older age at the time of surgery was predictive of better QuickDASH score and OES (P=.05 and P=.02, respectively). Patients who underwent surgery after a shorter duration of symptoms had better outcomes, but the difference did not reach statistical significance (QuickDASH, P=.09; OES, P=.10). Surgical treatment of recalcitrant medial epicondylitis with suture anchor fixation offers good pain relief and patient satisfaction, with little residual disability. Older age at the time of surgery predicts a better outcome.

  13. Geological significance of stone anchors from Dwarka waters, Gujarat, India

    Digital Repository Service at National Institute of Oceanography (India)

    Iyer, S.D.; Vora, K.H.; Gaur, A.S.

    as an offshore extension of routine, land based archaeology, the field of marine archaeology is increasing with advent of technology and interaction with other disciplines. of science, especially marine sciences. Aspects such as change of sea level...-ray Diffractometry (XRD) for identification of the mineral phases. A total of 122 anchors were collected of various shapes and sizes. The 25 rounded or ring stones (Pl. 42) are 30-70 cm in diameter, 34 triangular (PI. 43) ones are 180 cm to 55 cm long while...

  14. Anchoring a Lexicalized Tree-Adjoining Grammar for Discourse

    CERN Document Server

    Webber, Bryan R; Webber, Bonnie Lynn; Joshi, Aravind K.

    1998-01-01

    We here explore a ``fully'' lexicalized Tree-Adjoining Grammar for discourse that takes the basic elements of a (monologic) discourse to be not simply clauses, but larger structures that are anchored on variously realized discourse cues. This link with intra-sentential grammar suggests an account for different patterns of discourse cues, while the different structures and operations suggest three separate sources for elements of discourse meaning: (1) a compositional semantics tied to the basic trees and operations; (2) a presuppositional semantics carried by cue phrases that freely adjoin to trees; and (3) general inference, that draws additional, defeasible conclusions that flesh out what is conveyed compositionally.

  15. Anchor-based English-Chinese Bilingual Chunk Alignment Model

    Institute of Scientific and Technical Information of China (English)

    WU We-lin; CHENG Chang-sheng; XU Liang-xian; LU Ru-zhan

    2005-01-01

    Chunk alignment for the bilingual corpus is the base of Example-based Machine Translation. An anchor-based English-Chinese bilingual chunk alignment model and the corresponding algorithm of alignment are presented in this paper. It can effectively overcome the sparse data problem due to the limited size of the bilingual corpus. In this model, the chunk segmentation disambiguation is delayed to the alignment process, and hence the accuracy of chunk segmentation is improved. The experimental results demonstrate the feasibility and viability of this model.

  16. Results of Some Uplift Capacity Tests on Direct Embedment Anchors

    Science.gov (United States)

    1978-06-01

    should be expected to vary from those of the terrestrial tesis above. The analysis of direct embedment anchor-holding capacity is complicated by the...was conducted north of the Puerto Rico trench in 5,500 m (18,000 ft) in a pelagic clay or "red clay" deposit. Water contents in the upper 1.2 m (4 ft...north of Puerto Rico Trench. S6 - -~ ~-~d- 70. 60 20 60 j - 12 18 5 fluke 16 embedaent - 10 -0 340 6- -20 20,--6 4 - 10- ൟ 0g 30 2 3 4 5 6 7 4i Time

  17. Direct imaging of rotating molecules anchored on graphene

    Science.gov (United States)

    Choe, Jeongheon; Lee, Yangjin; Fang, Lei; Lee, Gun-Do; Bao, Zhenan; Kim, Kwanpyo

    2016-07-01

    There has been significant research interest in controlling and imaging molecular dynamics, such as translational and rotational motions, especially at a single molecular level. Here we applied aberration-corrected transmission electron microscopy (ACTEM) to actuate and directly image the rotational motions of molecules anchored on a single-layer-graphene sheet. Nanometer-sized carbonaceous molecules anchored on graphene provide ideal systems for monitoring rotational motions via ACTEM. We observed the preferential registry of longer molecular axis along graphene zigzag or armchair lattice directions due to the stacking-dependent molecule-graphene energy landscape. The calculated cross section from elastic scattering theory was used to experimentally estimate the rotational energy barriers of molecules on graphene. The observed energy barrier was within the range of 1.5-12 meV per atom, which is in good agreement with previous calculation results. We also performed molecular dynamics simulations, which revealed that the edge atoms of the molecule form stably bonds to graphene defects and can serve as a pivot point for rotational dynamics. Our study demonstrates the versatility of ACTEM for the investigation of molecular dynamics and configuration-dependent energetics at a single molecular level.There has been significant research interest in controlling and imaging molecular dynamics, such as translational and rotational motions, especially at a single molecular level. Here we applied aberration-corrected transmission electron microscopy (ACTEM) to actuate and directly image the rotational motions of molecules anchored on a single-layer-graphene sheet. Nanometer-sized carbonaceous molecules anchored on graphene provide ideal systems for monitoring rotational motions via ACTEM. We observed the preferential registry of longer molecular axis along graphene zigzag or armchair lattice directions due to the stacking-dependent molecule-graphene energy landscape. The

  18. Spectral Analysis of Surface Wave for Empirical Elastic Design of Anchored Foundations

    Directory of Open Access Journals (Sweden)

    S. E. Chen

    2012-01-01

    Full Text Available Helical anchors are vital support components for power transmission lines. Failure of a single anchor can lead to the loss of an entire transmission line structure which results in the loss of power for downstream community. Despite being important, it is not practical to use conventional borehole method of subsurface exploration, which is labor intensive and costly, for estimating soil properties and anchor holding capacity. This paper describes the use of an empirical and elasticity-based design technique coupled with the spectral analysis of surface wave (SASW technique to provide subsurface information for anchor foundation designs. Based on small-strain wave propagation, SASW determines shear wave velocity profile which is then correlated to anchor holding capacity. A pilot project involving over 400 anchor installations has been performed and demonstrated that such technique is reliable and can be implemented into transmission line structure designs.

  19. A novel role of Rab11 in trafficking GPI-anchored trans-sialidase to the plasma membrane of Trypanosoma cruzi.

    Science.gov (United States)

    Niyogi, Sayantanee; Docampo, Roberto

    2015-01-01

    Trypanosoma cruzi, the causative agent of Chagas disease, is a unicellular parasite that possesses a contractile vacuole complex (CVC). This organelle is usually present in free-living protists and is mainly involved in osmoregulation. However, in some organisms, like for example Dictyostelium discoideum, other roles include calcium homeostasis and transference of proteins to the plasma membrane. T. cruzi plasma membrane is very rich in glycosylphosphatidylinositol anchored proteins (GPI-AP) and a very important group of GPI-AP is that of the trans-sialidases. These enzymes catalyze the transfer of sialic acid from host glycoconjugates to mucins present in the surface of the parasite and are important for host cell invasion among other functions. We recently reported that a pathway dependent on the Rab GTPase Rab11 is involved in the traffic of trans-sialidases to the plasma membrane through the CVC of the infective stages of the parasite and that preventing this traffic results in considerable reduction in the ability of T. cruzi to infect host cells. We also found that traffic of other GPI-anchored proteins is also through the CVC but uses a Rab11-independent pathway. These represent unconventional pathways of GPI-anchored protein traffic to the plasma membrane.

  20. Displaying Lipase B from Candida antarctica in Pichia pastoris Using the Yeast Surface Display Approach: Prospection of a New Anchor and Characterization of the Whole Cell Biocatalyst

    Science.gov (United States)

    Moura, Marcelo Victor Holanda; da Silva, Giulia Pontes; Machado, Antônio Carlos de Oliveira; Torres, Fernando Araripe Gonçalves; Freire, Denise Maria Guimarães; Almeida, Rodrigo Volcan

    2015-01-01

    Yeast Surface Display (YSD) is a strategy to anchor proteins on the yeast cell wall which has been employed to increase enzyme stability thus decreasing production costs. Lipase B from Candida antarctica (LipB) is one of the most studied enzymes in the context of industrial biotechnology. This study aimed to assess the biochemical features of this important biocatalyst when immobilized on the cell surface of the methylotrophic yeast Pichia pastoris using the YSD approach. For that purpose, two anchors were tested. The first (Flo9) was identified after a prospection of the P. pastoris genome being related to the family of flocculins similar to Flo1 but significantly smaller. The second is the Protein with Internal Repeats (Pir1) from P. pastoris. An immunolocalization assay showed that both anchor proteins were able to display the reporter protein EGFP in the yeast outer cell wall. LipB was expressed in P. pastoris fused either to Flo9 (FLOLIPB) or Pir1 (PIRLIPB). Both constructions showed hydrolytic activity towards tributyrin (>100 U/mgdcw and >80 U/mgdcw, respectively), optimal hydrolytic activity around 45°C and pH 7.0, higher thermostability at 45°C and stability in organic solvents when compared to a free lipase. PMID:26510006

  1. Displaying Lipase B from Candida antarctica in Pichia pastoris Using the Yeast Surface Display Approach: Prospection of a New Anchor and Characterization of the Whole Cell Biocatalyst.

    Directory of Open Access Journals (Sweden)

    Marcelo Victor Holanda Moura

    Full Text Available Yeast Surface Display (YSD is a strategy to anchor proteins on the yeast cell wall which has been employed to increase enzyme stability thus decreasing production costs. Lipase B from Candida antarctica (LipB is one of the most studied enzymes in the context of industrial biotechnology. This study aimed to assess the biochemical features of this important biocatalyst when immobilized on the cell surface of the methylotrophic yeast Pichia pastoris using the YSD approach. For that purpose, two anchors were tested. The first (Flo9 was identified after a prospection of the P. pastoris genome being related to the family of flocculins similar to Flo1 but significantly smaller. The second is the Protein with Internal Repeats (Pir1 from P. pastoris. An immunolocalization assay showed that both anchor proteins were able to display the reporter protein EGFP in the yeast outer cell wall. LipB was expressed in P. pastoris fused either to Flo9 (FLOLIPB or Pir1 (PIRLIPB. Both constructions showed hydrolytic activity towards tributyrin (>100 U/mgdcw and >80 U/mgdcw, respectively, optimal hydrolytic activity around 45°C and pH 7.0, higher thermostability at 45°C and stability in organic solvents when compared to a free lipase.

  2. Displaying Lipase B from Candida antarctica in Pichia pastoris Using the Yeast Surface Display Approach: Prospection of a New Anchor and Characterization of the Whole Cell Biocatalyst.

    Science.gov (United States)

    Moura, Marcelo Victor Holanda; da Silva, Giulia Pontes; Machado, Antônio Carlos de Oliveira; Torres, Fernando Araripe Gonçalves; Freire, Denise Maria Guimarães; Almeida, Rodrigo Volcan

    2015-01-01

    Yeast Surface Display (YSD) is a strategy to anchor proteins on the yeast cell wall which has been employed to increase enzyme stability thus decreasing production costs. Lipase B from Candida antarctica (LipB) is one of the most studied enzymes in the context of industrial biotechnology. This study aimed to assess the biochemical features of this important biocatalyst when immobilized on the cell surface of the methylotrophic yeast Pichia pastoris using the YSD approach. For that purpose, two anchors were tested. The first (Flo9) was identified after a prospection of the P. pastoris genome being related to the family of flocculins similar to Flo1 but significantly smaller. The second is the Protein with Internal Repeats (Pir1) from P. pastoris. An immunolocalization assay showed that both anchor proteins were able to display the reporter protein EGFP in the yeast outer cell wall. LipB was expressed in P. pastoris fused either to Flo9 (FLOLIPB) or Pir1 (PIRLIPB). Both constructions showed hydrolytic activity towards tributyrin (>100 U/mgdcw and >80 U/mgdcw, respectively), optimal hydrolytic activity around 45°C and pH 7.0, higher thermostability at 45°C and stability in organic solvents when compared to a free lipase.

  3. Targeting and function of the mitochondrial fission factor GDAP1 are dependent on its tail-anchor.

    Directory of Open Access Journals (Sweden)

    Konstanze M Wagner

    Full Text Available Proteins controlling mitochondrial dynamics are often targeted to and anchored into the mitochondrial outer membrane (MOM by their carboxyl-terminal tail-anchor domain (TA. However, it is not known whether the TA modulates protein function. GDAP1 is a mitochondrial fission factor with two neighboring hydrophobic domains each flanked by basic amino acids (aa. Here we define GDAP1 as TA MOM protein. GDAP1 carries a single transmembrane domain (TMD that is, together with the adjacent basic aa, critical for MOM targeting. The flanking N-terminal region containing the other hydrophobic domain is located in the cytoplasm. TMD sequence, length, and high hydrophobicity do not influence GDAP1 fission function if MOM targeting is maintained. The basic aa bordering the TMD in the cytoplasm, however, are required for both targeting of GDAP1 as part of the TA and GDAP1-mediated fission. Thus, this GDAP1 region contains critical overlapping motifs defining intracellular targeting by the TA concomitant with functional aspects.

  4. Seismic Behavior and Modeling of Anchored Nonstructural Components Considering the Influence of Cyclic Cracks

    OpenAIRE

    Watkins, Derrick Andrew

    2011-01-01

    During an earthquake, reinforced concrete members in a building will suffer cracking that oscillates as the building dynamically deforms. Equipment that services the building, such as mechanical and electrical items, are anchored to these components, and therefore will be subjected to this dynamic environment. Despite understanding this practical loading situation, as well as recognizing that anchor load capacity is significantly reduced when an anchor is embedded in cracked concrete, there r...

  5. Effect of mutation of two critical glutamic acid residues on the activity and stability of human carboxypeptidase M and characterization of its signal for glycosylphosphatidylinositol anchoring.

    Science.gov (United States)

    Tan, Fulong; Balsitis, Scott; Black, Judy K; Blöchl, Andrea; Mao, Ji-Fang; Becker, Robert P; Schacht, David; Skidgel, Randal A

    2003-03-01

    Human carboxypeptidase (CP) M was expressed in baculovirus-infected insect cells in a glycosylphosphatidylinositol-anchored form, whereas a truncated form, lacking the putative signal sequence for glycosylphosphatidylinositol anchoring, was secreted at high levels into the medium. Both forms had lower molecular masses (50 kDa) than native placental CPM (62 kDa), indicating minimal glycosylation. The predicted glycosylphosphatidylinositol-anchor attachment site was investigated by mutation of Ser(406) to Ala, Thr or Pro and expression in HEK-293 and COS-7 cells. The wild-type and S406A and S406T mutants were expressed on the plasma membrane in glycosylphosphatidylinositol-anchored form, but the S406P mutant was not and was retained in a perinuclear location. The roles of Glu(260) and Glu(264) in CPM were investigated by site-directed mutagenesis. Mutation of Glu(260) to Gln had minimal effects on kinetic parameters, but decreased heat stability, whereas mutation to Ala reduced the k(cat)/ K(m) by 104-fold and further decreased stability. In contrast, mutation of Glu(264) to Gln resulted in a 10000-fold decrease in activity, but the enzyme still bound to p-aminobenzoylarginine-Sepharose and was resistant to trypsin treatment, indicating that the protein was folded properly. These results show that Glu(264) is the critical catalytic glutamic acid and that Glu(260) probably stabilizes the conformation of the active site.

  6. C-edge loops of arrestin function as a membrane anchor

    Science.gov (United States)

    Lally, Ciara C M.; Bauer, Brian; Selent, Jana; Sommer, Martha E

    2017-01-01

    G-protein-coupled receptors are membrane proteins that are regulated by a small family of arrestin proteins. During formation of the arrestin–receptor complex, arrestin first interacts with the phosphorylated receptor C terminus in a pre-complex, which activates arrestin for tight receptor binding. Currently, little is known about the structure of the pre-complex and its transition to a high-affinity complex. Here we present molecular dynamics simulations and site-directed fluorescence experiments on arrestin-1 interactions with rhodopsin, showing that loops within the C-edge of arrestin function as a membrane anchor. Activation of arrestin by receptor-attached phosphates is necessary for C-edge engagement of the membrane, and we show that these interactions are distinct in the pre-complex and high-affinity complex in regard to their conformation and orientation. Our results expand current knowledge of C-edge structure and further illuminate the conformational transitions that occur in arrestin along the pathway to tight receptor binding. PMID:28220785

  7. SNP discovery and chromosome anchoring provide the first physically-anchored hexaploid oat map and reveal synteny with model species.

    Directory of Open Access Journals (Sweden)

    Rebekah E Oliver

    Full Text Available A physically anchored consensus map is foundational to modern genomics research; however, construction of such a map in oat (Avena sativa L., 2n = 6x = 42 has been hindered by the size and complexity of the genome, the scarcity of robust molecular markers, and the lack of aneuploid stocks. Resources developed in this study include a modified SNP discovery method for complex genomes, a diverse set of oat SNP markers, and a novel chromosome-deficient SNP anchoring strategy. These resources were applied to build the first complete, physically-anchored consensus map of hexaploid oat. Approximately 11,000 high-confidence in silico SNPs were discovered based on nine million inter-varietal sequence reads of genomic and cDNA origin. GoldenGate genotyping of 3,072 SNP assays yielded 1,311 robust markers, of which 985 were mapped in 390 recombinant-inbred lines from six bi-parental mapping populations ranging in size from 49 to 97 progeny. The consensus map included 985 SNPs and 68 previously-published markers, resolving 21 linkage groups with a total map distance of 1,838.8 cM. Consensus linkage groups were assigned to 21 chromosomes using SNP deletion analysis of chromosome-deficient monosomic hybrid stocks. Alignments with sequenced genomes of rice and Brachypodium provide evidence for extensive conservation of genomic regions, and renewed encouragement for orthology-based genomic discovery in this important hexaploid species. These results also provide a framework for high-resolution genetic analysis in oat, and a model for marker development and map construction in other species with complex genomes and limited resources.

  8. SNP Discovery and Chromosome Anchoring Provide the First Physically-Anchored Hexaploid Oat Map and Reveal Synteny with Model Species

    Science.gov (United States)

    Chao, Shiaoman; Jellen, Eric N.; Carson, Martin L.; Rines, Howard W.; Obert, Donald E.; Lutz, Joseph D.; Shackelford, Irene; Korol, Abraham B.; Wight, Charlene P.; Gardner, Kyle M.; Hattori, Jiro; Beattie, Aaron D.; Bjørnstad, Åsmund; Bonman, J. Michael; Jannink, Jean-Luc; Sorrells, Mark E.; Brown-Guedira, Gina L.; Mitchell Fetch, Jennifer W.; Harrison, Stephen A.; Howarth, Catherine J.; Ibrahim, Amir; Kolb, Frederic L.; McMullen, Michael S.; Murphy, J. Paul; Ohm, Herbert W.; Rossnagel, Brian G.; Yan, Weikai; Miclaus, Kelci J.; Hiller, Jordan; Maughan, Peter J.; Redman Hulse, Rachel R.; Anderson, Joseph M.; Islamovic, Emir

    2013-01-01

    A physically anchored consensus map is foundational to modern genomics research; however, construction of such a map in oat (Avena sativa L., 2n = 6x = 42) has been hindered by the size and complexity of the genome, the scarcity of robust molecular markers, and the lack of aneuploid stocks. Resources developed in this study include a modified SNP discovery method for complex genomes, a diverse set of oat SNP markers, and a novel chromosome-deficient SNP anchoring strategy. These resources were applied to build the first complete, physically-anchored consensus map of hexaploid oat. Approximately 11,000 high-confidence in silico SNPs were discovered based on nine million inter-varietal sequence reads of genomic and cDNA origin. GoldenGate genotyping of 3,072 SNP assays yielded 1,311 robust markers, of which 985 were mapped in 390 recombinant-inbred lines from six bi-parental mapping populations ranging in size from 49 to 97 progeny. The consensus map included 985 SNPs and 68 previously-published markers, resolving 21 linkage groups with a total map distance of 1,838.8 cM. Consensus linkage groups were assigned to 21 chromosomes using SNP deletion analysis of chromosome-deficient monosomic hybrid stocks. Alignments with sequenced genomes of rice and Brachypodium provide evidence for extensive conservation of genomic regions, and renewed encouragement for orthology-based genomic discovery in this important hexaploid species. These results also provide a framework for high-resolution genetic analysis in oat, and a model for marker development and map construction in other species with complex genomes and limited resources. PMID:23533580

  9. Glucosamine-Anchored Graphene Oxide Nanosheets: Fabrication, Ultraviolet Irradiation, and Electrochemical Properties.

    Science.gov (United States)

    Veerapandian, Murugan; Lévaray, Nicolas; Lee, Min-Ho; Giasson, Suzanne; Zhu, X X

    2015-07-15

    A biofunctionalized graphene oxide (GO) nanosheet with improved physicochemical properties is useful for electrocatalysis and sensor development. Herein, a new class of functionalized GO with a chemically anchored biomolecule glucosamine is developed. Structural and chemical analyses confirm the glucosamine anchoring. Ultraviolet irradiation transforms the surface chemistry of GO. Glucosamine-anchored GO nanosheets exhibit improved cyclic voltammetric and amperometric sensing activity toward the model redox probe, ruthenium(II) and N-acetylneuraminic acid, respectively. The biomolecular anchoring and ultraviolet irradiation helped to tune and enhance the properties of GO, which may find multiple applications in optimizing sensor platforms.

  10. Optimization of Dead Load State in Earth-Anchored Cable-Stayed Bridges

    Institute of Scientific and Technical Information of China (English)

    Bin Sun∗; Rucheng Xiao

    2015-01-01

    In order to determine the reasonable completed dead load state in earth⁃anchored cable⁃stayed bridges, a practical method is proposed. The method is based on the rigidly supported continuous beam method and the feasible zone method, emphasizing on the mutual effect between the self⁃anchored structural parts and the earth⁃anchored ones. Three cable⁃stayed bridge models are designed with the main spans of 1 400 m, including a partially earth⁃anchored cable⁃stayed bridge, a cable⁃stayed⁃suspension bridge and a fully self⁃anchored cable⁃stayed bridge, in which the C50 concrete and Q345 steel are adopted. The partially earth⁃anchored cable⁃stayed bridge and the cable⁃stayed⁃suspension bridge secure lower compressive force in the girder than the fully self⁃anchored cable⁃stayed bridge by 25 percent at least. The same is for the material consumption of the whole bridge. Furthermore, the anchor volume is more than 20% lower in the partially earth⁃anchored cable⁃stayed bridge than that in the cable⁃stayed⁃suspension bridge. Consequently, the practical span of cable⁃stayed bridges can be accordingly extended.

  11. Suture anchor repair of quadriceps tendon rupture after total knee arthroplasty.

    Science.gov (United States)

    Kim, Tae Won B; Kamath, Atul F; Israelite, Craig L

    2011-08-01

    Disruption of the extensor mechanism after total knee arthroplasty (TKA) is a devastating complication, usually requiring surgical repair. Although suture anchor fixation is well described for repair of the ruptured native knee quadriceps tendon, no study has discussed the use of suture anchors in quadriceps repair after TKA. We present an illustrative case of successful suture anchor fixation of the quadriceps mechanism after TKA. The procedure has been performed in a total of 3 patients. A surgical technique and brief review of the literature follows. Suture anchor fixation of the quadriceps tendon is a viable option in the setting of rupture after TKA.

  12. Suture anchor versus suture through tunnel fixation for quadriceps tendon rupture: a biomechanical study.

    Science.gov (United States)

    Lighthart, William A; Cohen, David A; Levine, Richard G; Parks, Brent G; Boucher, Henry R

    2008-05-01

    This biomechanical study compared suture anchors versus transosseous sutures for repair of quadriceps tendon ruptures using a force of 150 N at a frequency of 0.5 Hz. No significant difference in displacement was found between the 2 techniques with initial loading or with load or no load after 1000 cycles. Displacement after 1000 cycles for suture anchors and bone tunnels was 4.65 and 4.50 mm, respectively. These findings suggest a possible role for suture anchors in repairing quadriceps tendon ruptures. Suture anchors are relatively expensive but require less dissection over the patella and do not involve suture placement about the patella tendon.

  13. Artificial Neural Network Model for Predicting Ultimate Tensile Capacity of Adhesive Anchors

    Institute of Scientific and Technical Information of China (English)

    XU Bo; WU Zhi-min; SONG Zhi-fei

    2007-01-01

    To predict the tensile capacity of adhesive anchors, a multilayered feed-forward neural network trained with the backpropagation algorithm is constructed. The ANN model have 5 inputs, including the compressive strength of concrete, tensile strength of concrete, anchor diameter, hole diameter, embedment of anchors, and ultimate load. The predictions obtained from the trained ANN show a good agreement with the experiments. Meanwhile, the predicted ultinate tensile capacity of anchors is close to the one calculated from the strength formula of the combined cone-bond failure model.

  14. Intermittent use of an "anchor system" improves postural control in healthy older adults.

    Science.gov (United States)

    Freitas, Milena de Bem Zavanella; Mauerberg-deCastro, Eliane; Moraes, Renato

    2013-07-01

    Haptic information, provided by a non-rigid tool (i.e., an "anchor system"), can reduce body sway in individuals who perform a standing postural task. However, it was not known whether or not continuous use of the anchor system would improve postural control after its removal. Additionally, it was unclear as to whether or not frequency of use of the anchor system is related to improved control in older adults. The present study evaluated the effect of the prolonged use of the anchor system on postural control in healthy older individuals, at different frequencies of use, while they performed a postural control task (semi-tandem position). Participants were divided into three groups according to the frequency of the anchor system's use (0%, 50%, and 100%). Pre-practice phase (without anchor) was followed by a practice phase (they used the anchor system at the predefined frequency), and a post-practice phase (immediate and late-without anchor). All three groups showed a persistent effect 15min after the end of the practice phase (immediate post-practice phase). However, only the 50% group showed a persistent effect in the late post-practice phase (24h after finishing the practice phase). Older adults can improve their postural control by practicing the standing postural task, and use of the anchor system limited to half of their practice time can provide additional improvement in their postural control.

  15. Post-installed concrete anchors in nuclear power plants: Performance and qualification

    Energy Technology Data Exchange (ETDEWEB)

    Mahrenholtz, Philipp, E-mail: philipp@mahrenholtz.net; Eligehausen, Rolf

    2015-06-15

    Graphical abstract: - Highlights: • Review of qualification and design regulations for anchors in nuclear power plants. • First complete set of nuclear anchor load–displacement data and its evaluation ever. • Demonstration of robust test behavior of a qualified post-installed anchor product. - Abstract: In nuclear power plants (NPPs), post-installed anchors are widely used for structural and non-structural connections to concrete. In many countries, anchor products employed for safety relevant applications have to be approved by the authorities. For the high safety standards in force for NPPs, special requirements have to be met to allow for extreme design situations. This paper presents an experimental test program conducted to evaluate the performance of anchors according to the German Guideline for Anchorages in Nuclear Power Plants and Nuclear Technology Installations (DIBt KKW Leitfaden, 2010). After a brief introduction to anchor behavior and the regulative context, the results of tension and shear tests carried out on undercut anchors are discussed. Robust load capacities and relatively small displacements determined for demanding load and crack cycling tests demonstrated the suitability of anchors qualified according to a state-of-the-art qualification guideline.

  16. Sialic Acid on the Glycosylphosphatidylinositol Anchor Regulates PrP-mediated Cell Signaling and Prion Formation.

    Science.gov (United States)

    Bate, Clive; Nolan, William; Williams, Alun

    2016-01-01

    The prion diseases occur following the conversion of the cellular prion protein (PrP(C)) into disease-related isoforms (PrP(Sc)). In this study, the role of the glycosylphosphatidylinositol (GPI) anchor attached to PrP(C) in prion formation was examined using a cell painting technique. PrP(Sc) formation in two prion-infected neuronal cell lines (ScGT1 and ScN2a cells) and in scrapie-infected primary cortical neurons was increased following the introduction of PrP(C). In contrast, PrP(C) containing a GPI anchor from which the sialic acid had been removed (desialylated PrP(C)) was not converted to PrP(Sc). Furthermore, the presence of desialylated PrP(C) inhibited the production of PrP(Sc) within prion-infected cortical neurons and ScGT1 and ScN2a cells. The membrane rafts surrounding desialylated PrP(C) contained greater amounts of sialylated gangliosides and cholesterol than membrane rafts surrounding PrP(C). Desialylated PrP(C) was less sensitive to cholesterol depletion than PrP(C) and was not released from cells by treatment with glimepiride. The presence of desialylated PrP(C) in neurons caused the dissociation of cytoplasmic phospholipase A2 from PrP-containing membrane rafts and reduced the activation of cytoplasmic phospholipase A2. These findings show that the sialic acid moiety of the GPI attached to PrP(C) modifies local membrane microenvironments that are important in PrP-mediated cell signaling and PrP(Sc) formation. These results suggest that pharmacological modification of GPI glycosylation might constitute a novel therapeutic approach to prion diseases.

  17. Bone anchor systems for orthodontic application: a systematic review.

    Science.gov (United States)

    Tsui, W K; Chua, H D P; Cheung, L K

    2012-11-01

    This systematic review was performed to investigate the usefulness and clinical effectiveness of skeletal anchorage devices to determine the most effective bone anchor system for orthodontic tooth movement. Literature on bone anchorage devices was selected from PubMed and the Cochrane Library from January 1966 to June 2010. 55 publications regarding miniplates, miniscrews, palatal implants and dental implants as orthodontic anchorage were identified for further analysis. All bone anchorage devices were found to have relatively high success rates and demonstrated their ability to provide absolute anchorage for orthodontic tooth movement. Significant tooth movement could be achieved with low morbidities and good patient acceptance. The reported success rates for the four groups of anchorage systems were generally high with slight variability (miniplates 91.4-100%; palatal implants 74-93.3%; miniscrews 61-100%; dental implants 100%). It was concluded that bone anchorage systems can achieve effective orthodontic movement with low morbidities. The success rate is generally high with slight variability between miniplates, palatal implants, miniscrews and dental implants. Owing to the lack of randomized controlled trials, there is no strong evidence to confirm which bone anchor system is the most effective for orthodontic tooth movement.

  18. Cognitive Advantages of Blending with Material Anchors in Energy Instruction

    Science.gov (United States)

    Close, Hunter; Close, Eleanor; Scherr, Rachel; McKagan, Sarah

    2012-03-01

    Conceptual blending theory [1] explains how the human imagination creates unreal situations that help us think about reality. In these imaginary blended situations, we establish new correspondences, interactions, and dynamics, and the outcomes of the dynamics lend insight to the nature of various real situations that were used to compose the blend. Blends are not just in the head, however; in some cases, a material system participates in the blend by lending its material structure as conceptual structure [2]. In the instructional activity Energy Theater [3], people represent units of energy and move around in order to solve puzzles of energy transfer and transformation. We use the ideas of blending and material anchors to understand how learners are able to use the representation to their cognitive advantage. [4pt] [1] Fauconnier, G. & Turner, M. (2002). The Way We Think: Conceptual Blending and the Mind's Hidden Complexities. New York: Basic Books.[0pt] [2] Hutchins, E. (2005) Material anchors for conceptual blends. Journal of Pragmatics 37, 1555-1577.[0pt] [3] Scherr, R. E., Close, H. G., McKagan, S. B., & Close, E. W. (2010) ``Energy Theater'': Using the body symbolically to understand energy. In C. Singh, M. Sabella, & S. Rebello (Eds.) 2010 PERC Proceedings. Melville, NY: AIP Press.

  19. Glycosyl-phosphatidylinositol (GPI)-anchored membrane association of the porcine reproductive and respiratory syndrome virus GP4 glycoprotein and its co-localization with CD163 in lipid rafts

    Energy Technology Data Exchange (ETDEWEB)

    Du, Yijun [Department of Pathobiology, University of Illinois at Urbana-Champaign, 2001 South Lincoln Ave, Urbana, IL 61802 (United States); Shandong Key Laboratory of Animal Disease Control and Breeding, Institute of Animal Science and Veterinary Medicine, Shandong Academy of Agricultural Sciences, Jinan (China); Pattnaik, Asit K. [School of Veterinary Medicine and Biomedical Sciences and the Nebraska Center for Virology, University of Nebraska-Lincoln, Lincoln, NE 68583-0900 (United States); Song, Cheng [Department of Pathobiology, University of Illinois at Urbana-Champaign, 2001 South Lincoln Ave, Urbana, IL 61802 (United States); Yoo, Dongwan, E-mail: dyoo@illinois.edu [Department of Pathobiology, University of Illinois at Urbana-Champaign, 2001 South Lincoln Ave, Urbana, IL 61802 (United States); Li, Gang, E-mail: dyoo@illinois.edu [Department of Pathobiology, University of Illinois at Urbana-Champaign, 2001 South Lincoln Ave, Urbana, IL 61802 (United States); Institute of Animal Science and Veterinary Medicine, Chinese Academy of Agricultural Sciences, Beijing (China)

    2012-03-01

    The porcine reproductive and respiratory syndrome virus (PRRSV) glycoprotein 4 (GP4) resembles a typical type I membrane protein in its structure but lacks a hydrophilic tail at the C-terminus, suggesting that GP4 may be a lipid-anchored membrane protein. Using the human decay-accelerating factor (DAF; CD55), a known glycosyl-phosphatidylinositol (GPI) lipid-anchored protein, chimeric constructs were made to substitute the GPI-anchor domain of DAF with the putative lipid-anchor domain of GP4, and their membrane association and lipase cleavage were determined in cells. The DAF-GP4 fusion protein was transported to the plasma membrane and was cleaved by phosphatidylinositol-specific phospholipase C (PI-PLC), indicating that the C-terminal domain of GP4 functions as a GPI anchor. Mutational studies for residues adjacent to the GPI modification site and characterization of respective mutant viruses generated from infectious cDNA clones show that the ability of GP4 for membrane association corresponded to virus viability and growth characteristics. The residues T158 ({omega} - 2, where {omega} is the GPI moiety at E160), P159 ({omega} - 1), and M162 ({omega} + 2) of GP4 were determined to be important for virus replication, with M162 being of particular importance for virus infectivity. The complete removal of the peptide-anchor domain in GP4 resulted in a complete loss of virus infectivity. The depletion of cholesterol from the plasma membrane of cells reduced the virus production, suggesting a role of lipid rafts in PRRSV infection. Remarkably, GP4 was found to co-localize with CD163 in the lipid rafts on the plasma membrane. Since CD163 has been reported as a cellular receptor for PRRSV and GP4 has been shown to interact with this receptor, our data implicates an important role of lipid rafts during entry of the virus.

  20. 'Saurashtra stone anchors' (Ring-stones) from Dwarka and Somnath, west coast of India

    Digital Repository Service at National Institute of Oceanography (India)

    Gaur, A.S.; Sundaresh; Tripati, S.; Gudigar, P.; Bandodkar, S.N.

    , Vijaydurg and Sindhudurg in Maharashtra. A few grapnel-type stone anchors have been reported from Lakshadweep and from Tamil Nadu coast. The majority of stone anchors from Gujarat fall in three categories: (1) composite, (2) grapnel, and (3) ring stone types...

  1. An Anchor-Based Pedestrian Navigation Approach Using Only Inertial Sensors.

    Science.gov (United States)

    Gu, Yang; Song, Qian; Li, Yanghuan; Ma, Ming; Zhou, Zhimin

    2016-03-07

    In inertial-based pedestrian navigation, anchors can effectively compensate the positioning errors originating from deviations of Inertial Measurement Units (IMUs), by putting constraints on pedestrians' motions. However, these anchors often need to be deployed beforehand, which can greatly increase system complexity, rendering it unsuitable for emergency response missions. In this paper, we propose an anchor-based pedestrian navigation approach without any additional sensors. The anchors are defined as the intersection points of perpendicular corridors and are considered characteristics of building structures. In contrast to these real anchors, virtual anchors are extracted from the pedestrian's trajectory and are considered as observations of real anchors, which can accordingly be regarded as inferred building structure characteristics. Then a Rao-Blackwellized particle filter (RBPF) is used to solve the joint estimation of positions (trajectory) and maps (anchors) problem. Compared with other building structure-based methods, our method has two advantages. The assumption on building structure is minimum and valid in most cases. Even if the assumption does not stand, the method will not lead to positioning failure. Several real-scenario experiments are conducted to validate the effectiveness and robustness of the proposed method.

  2. An Anchor-Based Pedestrian Navigation Approach Using Only Inertial Sensors

    Directory of Open Access Journals (Sweden)

    Yang Gu

    2016-03-01

    Full Text Available In inertial-based pedestrian navigation, anchors can effectively compensate the positioning errors originating from deviations of Inertial Measurement Units (IMUs, by putting constraints on pedestrians’ motions. However, these anchors often need to be deployed beforehand, which can greatly increase system complexity, rendering it unsuitable for emergency response missions. In this paper, we propose an anchor-based pedestrian navigation approach without any additional sensors. The anchors are defined as the intersection points of perpendicular corridors and are considered characteristics of building structures. In contrast to these real anchors, virtual anchors are extracted from the pedestrian’s trajectory and are considered as observations of real anchors, which can accordingly be regarded as inferred building structure characteristics. Then a Rao-Blackwellized particle filter (RBPF is used to solve the joint estimation of positions (trajectory and maps (anchors problem. Compared with other building structure-based methods, our method has two advantages. The assumption on building structure is minimum and valid in most cases. Even if the assumption does not stand, the method will not lead to positioning failure. Several real-scenario experiments are conducted to validate the effectiveness and robustness of the proposed method.

  3. Career Anchors: Distribution and Impact on Job Satisfaction, the Israeli Case

    Science.gov (United States)

    Danziger, Nira; Valency, Rony

    2006-01-01

    Purpose: The purpose of this paper is to examine the career anchor concept developed by Edgar Schein. Design/methodology/approach: The paper focuses on the distribution of the eight career anchors, on a large heterogeneous sample and the differences in the distribution by gender and type of employment; and the impact of the congruence on job…

  4. MODIFICATION OF PALLADIUM METALLIC CATALYST WITH POLYMER-ANCHORED THIOETHER LIGANDS

    Institute of Scientific and Technical Information of China (English)

    LIU Hanfan; MAO Guoping

    1993-01-01

    A well-dispersed metallic palladium catalyst modified by polymer-anchored thioether ligands was used for the hydrogenation of cyclopentadiene to cyclopentene with high activity and selectivity in ambient condition. The evidences to show the modification of catalytic properties by polymer anchored ligands were given.

  5. Origins of the Stuttering Stereotype: Stereotype Formation through Anchoring-Adjustment

    Science.gov (United States)

    MacKinnon, Sean P.; Hall, Shera; MacIntyre, Peter D.

    2007-01-01

    The stereotype of people who stutter is predominantly negative, holding that stutterers are excessively nervous, anxious, and reserved. The anchoring-adjustment hypothesis suggests that the stereotype of stuttering arises from a process of first anchoring the stereotype in personal feelings during times of normal speech disfluency, and then…

  6. Effects of Anchored Instruction on Enhancing Chinese Students' Problem-Solving Skills.

    Science.gov (United States)

    Shyu, Hsin-Yih

    This study investigates the effects of computer-aided videodisc-based anchored instruction on promoting elementary school students' problem-solving skills in Taiwan. Anchored instruction combines theories such as situated cognition, cognitive apprenticeship, cooperative learning, and constructivist theories. With the help of interactive videodisc…

  7. Using Video-Based Anchored Instruction To Enhance Learning: Taiwan's Experience.

    Science.gov (United States)

    Shyu, Hsin-Yih Cindy

    2000-01-01

    Describes a study that investigated the effects of computer-assisted videodisc-based anchored instruction on attitudes toward mathematics and instruction as well as problem-solving skills among Taiwanese elementary students. Findings suggest that video-based anchored instruction provided a more motivating environment that enhanced students'…

  8. An Exploratory Comparison of Traditional Classroom Instruction and Anchored Instruction with Secondary School Students: Turkish Experience

    Science.gov (United States)

    Elcin, Melih; Sezer, Baris

    2014-01-01

    The purpose of this study was to investigate the impact of anchored instruction on the students in secondary school math studies classrooms. This study adopted a quasi-experimental design. This research involved both quantitative and qualitative methods to investigate the effects of anchored instruction on students' academical achievement,…

  9. A Comparison of Traditional Classroom Instruction and Anchored Instruction with University General Education Students.

    Science.gov (United States)

    Langone, John; Malone, D. Michael; Stecker, Pamela M.; Greene, Eric

    1998-01-01

    Effects of a traditional instructional format and technology-enhanced anchored instruction on the immediate and long-term acquisition of knowledge was evaluated with 100 university students in a special-education course. The CD-ROM-based anchored-instruction group outperformed the traditional instruction group on the multiple-choice follow-up test…

  10. Anchored Instruction: A Model for Integrating the Language Arts through Content Area Study.

    Science.gov (United States)

    Cena, Michael E.; Mitchell, Judith P.

    1998-01-01

    Presents a description of anchored instruction, a model of curriculum integration combined with inquiry learning that builds prior knowledge and engages students in the application of the language arts. Sets out the eight steps involved in creating an anchored instruction unit. (SR)

  11. The Effects of Technology-Enhanced Anchored Instruction on the Knowledge of Preservice Special Educators.

    Science.gov (United States)

    Langone, John; Malone, D. Michael; Clinton, Gregory N.

    1999-01-01

    The immediate and long-term acquisition of knowledge of 37 students divided into two groups (nonanchored instruction and anchored instruction) was explored. Although no differences between the two groups on the posttest immediately following the lectures was noted, the anchored instruction group outperformed the nonanchored group on the eight-week…

  12. Preliminary design and parametric study of 1400 m partially earth-anchored cable-stayed bridge

    Institute of Scientific and Technical Information of China (English)

    2010-01-01

    The preliminary design and parametric study of 1400 m partially earth-anchored cable-stayed bridge are described. Static per-formance of this new type of bridge is discussed. Compared with fully self-anchored cable-stayed bridge, its advantages in fin-ished state are summarized. Based on numerical calculation, effects of several structural parameters on static performance are presented.

  13. Anchoring and Estimation of Alcohol Consumption: Implications for Social Norm Interventions

    Science.gov (United States)

    Lombardi, Megan M.; Choplin, Jessica M.

    2010-01-01

    Three experiments investigated the impact of anchors on students' estimates of personal alcohol consumption to better understand the role that this form of bias might have in social norm intervention programs. Experiments I and II found that estimates of consumption were susceptible to anchoring effects when an open-answer and a scale-response…

  14. 33 CFR 401.49 - Dropping anchor or tying to canal bank.

    Science.gov (United States)

    2010-07-01

    ... 33 Navigation and Navigable Waters 3 2010-07-01 2010-07-01 false Dropping anchor or tying to canal bank. 401.49 Section 401.49 Navigation and Navigable Waters SAINT LAWRENCE SEAWAY DEVELOPMENT CORPORATION, DEPARTMENT OF TRANSPORTATION SEAWAY REGULATIONS AND RULES Regulations Seaway Navigation § 401.49 Dropping anchor or tying to canal...

  15. Anchoring Effects on Prospective and Retrospective Metacomprehension Judgments as a Function of Peer Performance Information

    Science.gov (United States)

    Zhao, Qin; Linderholm, Tracy

    2011-01-01

    Two experiments were conducted to investigate anchoring effects on metacomprehension judgments as a function of fictitious information participants received about past peer performance. In Experiment 1 participants were randomly assigned to one of the three anchor groups that, in some cases, provided past peer performance averages in terms of a…

  16. The use of suture anchors to repair the ruptured quadriceps tendon.

    Science.gov (United States)

    Bushnell, Brandon D; Whitener, George B; Rubright, James H; Creighton, R Alexander; Logel, Kevin J; Wood, Mark L

    2007-07-01

    Quadriceps tendon rupture is an incapacitating injury that usually requires surgical repair. Traditional repair methods involve transpatellar suture tunnels, but recent reports have introduced the idea of using suture anchors to repair the ruptured tendon. We present 5 cases of our technique of using suture anchors to repair the ruptured quadriceps tendon.

  17. An Exploratory Comparison of Traditional Classroom Instruction and Anchored Instruction with Secondary School Students: Turkish Experience

    Science.gov (United States)

    Elcin, Melih; Sezer, Baris

    2014-01-01

    The purpose of this study was to investigate the impact of anchored instruction on the students in secondary school math studies classrooms. This study adopted a quasi-experimental design. This research involved both quantitative and qualitative methods to investigate the effects of anchored instruction on students' academical achievement,…

  18. Rapid Naming Deficits in Dyslexia: A Stumbling Block for the Perceptual Anchor Theory of Dyslexia

    Science.gov (United States)

    Di Filippo, Gloria; Zoccolotti, Pierluigi; Ziegler, Johannes C.

    2008-01-01

    According to a recent theory of dyslexia, the "perceptual anchor theory," children with dyslexia show deficits in classic auditory and phonological tasks not because they have auditory or phonological impairments but because they are unable to form a "perceptual anchor" in tasks that rely on a small set of repeated stimuli. The theory makes the…

  19. Stone anchors from Sindhudurg Fort on the west coast of India

    Digital Repository Service at National Institute of Oceanography (India)

    Tripati, S.; Gaur, A.S.

    , of which there are triangular and three grapnet-type, used to construct the jetty along with the locally available sand stone. The two remaining grapnet-type anchors are erected on the jetty for mooring purposes. It is found that none of the anchors were...

  20. Immunogenic Eimeria tenella glycosylphosphatidylinositol-anchored surface antigens (SAGs induce inflammatory responses in avian macrophages.

    Directory of Open Access Journals (Sweden)

    Yock-Ping Chow

    Full Text Available BACKGROUND: At least 19 glycosylphosphatidylinositol (GPI-anchored surface antigens (SAGs are expressed specifically by second-generation merozoites of Eimeria tenella, but the ability of these proteins to stimulate immune responses in the chicken is unknown. METHODOLOGY/PRINCIPAL FINDINGS: Ten SAGs, belonging to two previously defined multigene families (A and B, were expressed as soluble recombinant (r fusion proteins in E. coli. Chicken macrophages were treated with purified rSAGs and changes in macrophage nitrite production, and in mRNA expression profiles of inducible nitric oxide synthase (iNOS and of a panel of cytokines were measured. Treatment with rSAGs 4, 5, and 12 induced high levels of macrophage nitric oxide production and IL-1β mRNA transcription that may contribute to the inflammatory response observed during E. tenella infection. Concomitantly, treatment with rSAGs 4, 5 and 12 suppressed the expression of IL-12 and IFN-γ and elevated that of IL-10, suggesting that during infection these molecules may specifically impair the development of cellular mediated immunity. CONCLUSIONS/SIGNIFICANCE: In summary, some E. tenella SAGs appear to differentially modulate chicken innate and humoral immune responses and those derived from multigene family A (especially rSAG 12 may be more strongly linked with E. tenella pathogenicity associated with the endogenous second generation stages.

  1. Different size biomolecules anchoring on porous silicon surface: fluorescence and reflectivity pores infiltration comparative studies

    Energy Technology Data Exchange (ETDEWEB)

    Giovannozzi, Andrea M.; Rossi, Andrea M. [National Institute for Metrological Research, Thermodynamic Division, Strada delle Cacce 91, 10135 Torino (Italy); Renacco, Chiara; Farano, Alessandro [Ribes Ricecrhe Srl, Via Lavoratori Vittime del Col du Mont 24, 11100 Aosta (Italy); Derosas, Manuela [Biodiversity Srl, Via Corfu 71, 25124 Brescia (Italy); Enrico, Emanuele [National Institute for Metrological Research, Electromagnetism Division, Strada delle Cacce 91, 10135 Torino (Italy)

    2011-06-15

    The performance of porous silicon optical based biosensors strongly depends on material nanomorphology, on biomolecules distribution inside the pores and on the ability to link sensing species to the pore walls. In this paper we studied the immobilization of biomolecules with different size, such as antibody anti aflatoxin (anti Aflatox Ab, {proportional_to}150 KDa), malate dehydrogenase (MDH, {proportional_to}36KDa) and metallothionein (MT, {proportional_to}6KDa) at different concentrations on mesoporous silicon samples ({proportional_to}15 nm pores diameter). Fluorescence measurements using FITC- labeled biomolecules and refractive index analysis based on reflectivity spectra have been employed together to detect the amount of proteins bound to the surface and to evaluate their diffusion inside the pores. Here we suggest that these two techniques should be used together to have a better understanding of what happens at the porous silicon surface. In fact, when pores dimensions are not perfectly tuned to the protein size a higher fluorescence signal doesn't often correspond to a higher biomolecules distribution inside the pores. When a too much higher concentration of biomolecule is anchored on the surface, steric crowd effects and repulsive interactions probably take over and hinder pores infiltration, inducing a small or absent shift in the fringe pattern even if a higher fluorescence signal is registered. (copyright 2011 WILEY-VCH Verlag GmbH and Co. KGaA, Weinheim) (orig.)

  2. Selective Removal of Hemoglobin from Blood Using Hierarchical Copper Shells Anchored to Magnetic Nanoparticles

    Science.gov (United States)

    Wang, Yaokun; Yan, Mingyang

    2017-01-01

    Hierarchical copper shells anchored on magnetic nanoparticles were designed and fabricated to selectively deplete hemoglobin from human blood by immobilized metal affinity chromatography. Briefly, CoFe2O4 nanoparticles coated with polyacrylic acid were first synthesized by a one-pot solvothermal method. Hierarchical copper shells were then deposited by immobilizing Cu2+ on nanoparticles and subsequently by reducing between the solid CoFe2O4@COOH and copper solution with NaBH4. The resulting nanoparticles were characterized by scanning electron microscopy, transmission electron microscopy, Fourier transform infrared spectrometry, X-ray photoelectron spectroscopy, and vibrating sample magnetometry. The particles were also tested against purified bovine hemoglobin over a range of pH, contact time, and initial protein concentration. Hemoglobin adsorption followed pseudo-second-order kinetics and reached equilibrium in 90 min. Isothermal data also fit the Langmuir model well, with calculated maximum adsorption capacity 666 mg g−1. Due to the high density of Cu2+ on the shell, the nanoparticles efficiently and selectively deplete hemoglobin from human blood. Taken together, the results demonstrate that the particles with hierarchical copper shells effectively remove abundant, histidine-rich proteins, such as hemoglobin from human blood, and thereby minimize interference in diagnostic and other assays.

  3. [Refixation of sternoclavicular luxation with a suture anchor system].

    Science.gov (United States)

    Lehmann, W; Laskowski, J; Grossterlinden, L; Rueger, J M

    2010-05-01

    Luxations of the sternoclavicular joint are rare injuries. We present a case of anterior dislocation which was caused by a minor fall onto the right shoulder. Diagnosis was delayed by 2 weeks which prevented treatment by closed reduction. As an alternative surgical treatment a PDS cord around the clavicle and the first rib was used to stabilize the clavicle at the vertical level. Subsequently, fixation to the medial side was achieved by a suture anchor that was placed into the manubrium. The presented case highlights this simple and safe method to treat dislocations of the sternoclavicular joint in which standard treatment cannot be performed. A detailed description for each operation step is given and our experience in terms of aftercare and outcome is reported.

  4. Codend selectivity in a commercial Danish anchor seine

    DEFF Research Database (Denmark)

    Noack, Thomas; Frandsen, Rikke; Krag, Ludvig Ahm

    2017-01-01

    Danish seining (or anchor seining) is a fishing technique that is gaining increasing attention because itis considered to be a fuel-efficient fishing method with low environmental impact. However, scientificdocumentation of the selectivity characteristics of Danish seines is lacking, and the gear...... generally isgrouped with bottom trawls and Scottish seines in fisheries management legislation. In this study, wedeveloped a codend cover to estimate the selectivity of a standard commercial Danish seine codendfor four fish species. The data for the dominant species, dab (Limanda limanda) and plaice...... (Pleuronectesplatessa), was best described by models that combine two or three logistic models, which indicated thatmore than one selection process was at work. Selectivity of cod (Gadus morhua) was best described by aRichard curve and selectivity of red gurnard (Chelidonichthys lucernus) by a logistic curve...

  5. Ideological Differences in Anchoring and Adjustment During Social Inferences.

    Science.gov (United States)

    Stern, Chadly; West, Tessa V

    2016-09-08

    Recent research has demonstrated that conservatives perceive greater similarity to political ingroup members than do liberals. In two studies, we draw from a framework of "anchoring and adjustment" to understand why liberals and conservatives differ in their perceptions of ingroup similarity. Results indicate that when participants made judgments under time pressure, liberals and conservatives did not differ in assuming ingroup similarity. However, when participants were given sufficient time to make judgments, liberals assumed less similarity than conservatives did, suggesting that liberals adjusted their judgments to a greater extent than conservatives did (Studies 1 and 2). In examining an underlying motivational process, we found that when conservatives' desire to affiliate with others was attenuated, they adjusted their initial judgments of ingroup similarity to a similar extent as liberals did (Study 2). We discuss implications for research on ideology and social judgment.

  6. Robust conductance of dumbbell molecular junctions with fullerene anchoring groups

    DEFF Research Database (Denmark)

    Markussen, Troels; Settnes, Mikkel; Thygesen, Kristian Sommer

    2011-01-01

    The conductance of a molecular wire connected to metallic electrodes is known to be sensitive to the atomic structure of the molecule-metal contact. This contact is to a large extent determined by the anchoring group linking the molecular wire to the metal. It has been found experimentally...... support the original motivation to achieve conductance values more stable towards changes in the structure of the molecule-metal contact leading to larger reproducibility in experiments. © 2011 American Institute of Physics....... against variations in the detailed bonding geometry and in good agreement with the experimental value of G= 3 × 10-4G0. Electron tunneling across the molecular bridge occurs via the lowest unoccupied orbitals of C60 which are pinned close to the Fermi energy due to partial charge transfer. Our findings...

  7. Anchoring FRP Composite Armor in Flexible Offshore Riser Systems

    DEFF Research Database (Denmark)

    Costache, Andrei

    capabilities.The structure of flexible pipes consists of several concentric layers, each with a specific purpose. The most common used flexible pipe is the type III, which contains a central component, made from an interlocking stainless steel structure that provides collapse strength. The central component...... armor layer is made from steel. However, as oil exploitation goes to deeper and deeper waters, the strength/weight ratio of steel armor become sunfavorable. In order to achieve higher tensile strength and to reduce the overall weight of the pipe, in the future, the tensile armor must be made...... of composite materials. One of the problems related to the substitution of tensile steel members is that anchoring in the metallic end fittings of the pipe is very challenging.The purpose of this thesis is to ensure the transfer of tensile loads between a unidirectional fiber reinforced polymer and a metallic...

  8. Anchor cell invasion into the vulval epithelium in C. elegans.

    Science.gov (United States)

    Sherwood, David R; Sternberg, Paul W

    2003-07-01

    An understanding of cell-invasive behavior has been limited by the lack of in vivo models where this activity can be clearly visualized and manipulated. We show that a single cell in the Caenorhabditis elegans gonad, the anchor cell (AC), initiates uterine-vulval contact through a cell invasion event. Using genetic analysis, laser ablations, and cell-specific markers, we demonstrate that AC invasion is predominantly stimulated by the 1 degrees vulval lineage cells, which generate a diffusible signal that promotes AC invasive behavior toward these cells and further targets invasive processes between the two central 1 degrees vulval lineage cells. We also show that AC invasion is regulated by the AC response to this cue, as well as a vulval-independent mechanism that weakly drives invasion. These studies dissect the regulatory mechanisms that underlie a simple cell-invasive behavior in vivo, and introduce AC invasion as a model for understanding key checkpoints controlling cell invasion.

  9. Smos Land Product Validation Activities at the Valencia Anchor Station

    Science.gov (United States)

    Lopez-Baeza, Ernesto

    ABSTRACT Soil moisture is a key parameter controlling the exchanges between the land surface and the atmosphere. In spite of being important for weather and climate modeling, this parameter is not well observed at a global scale. The SMOS (Soil Moisture and Ocean Salinity) Mission was designed by the European Space Agency (ESA) to measure soil moisture over continental surfaces as well as surface salinity over the oceans. Since 2001, the Valencia Anchor Station is currently being prepared for the validation of SMOS land products, namely soil moisture content and vegetation water content. The site has recently been selected by the Mission as a core validation site, mainly due to the reasonable homogeneous characteristics of the area which make it appropriate to undertake the validation of SMOS Level 2 land products during the Mission Commissioning Phase, before attempting more complex areas. Close to SMOS launch, ESA has defined and designed a SMOS V alidation Rehearsal C ampaign P lan which purpose is to repeat the Commissioning Phase execution with all centers, all tools, all participants, all structures, all data available, assuming all tools and structures are ready and trying to produce as close as possible the post-launch conditions. The aim is to test the readiness, the ensemble coordination and the speed of operations, and to avoid as far as possible any unexpected deficiencies of the plan and procedure during the real C ommissioning P hase campaigns. For the rehearsal activity, a control area of 10 x 10 km2 has been chosen at the Valencia Anchor Station study area where a network of ground soil moisture measuring stations is being set up based on the definition of homogeneous physio-hydrological units, attending to climatic, soil type, lithology, geology, elevation, slope and vegetation cover conditions. These stations are linked via a wireless communication system to a master post accessible via internet. The ground soil moisture stations will also be used

  10. The use of a neutral peptide aptamer scaffold to anchor BH3 peptides constitutes a viable approach to studying their function.

    Science.gov (United States)

    Stadler, L K J; Tomlinson, D C; Lee, T; Knowles, M A; Ko Ferrigno, P

    2014-01-30

    The B-cell CLL/lymphoma-2 (Bcl-2) family of proteins are important regulators of the intrinsic pathway of apoptosis, and their interactions, driven by Bcl-2 homology (BH) domains, are of great interest in cancer research. Particularly, the BH3 domain is of clinical relevance, as it promotes apoptosis through activation of Bcl-2-associated x protein (Bax) and Bcl-2 antagonist killer (Bak), as well as by antagonising the anti-apoptotic Bcl-2 family members. Although investigated extensively in vitro, the study of the BH3 domain alone inside cells is more problematic because of diminished secondary structure of the unconstrained peptide and a lack of stability. In this study, we report the successful use of a novel peptide aptamer scaffold - Stefin A quadruple mutant - to anchor and present the BH3 domains from Bcl-2-interacting mediator of cell death (Bim), p53 upregulated modulator of apoptosis (Puma), Bcl-2-associated death promoter (Bad) and Noxa, and demonstrate its usefulness in the study of the BH3 domains in vivo. When expressed intracellularly, anchored BH3 peptides exhibit much the same binding specificities previously established in vitro, however, we find that, at endogenous expression levels, Bcl-2 does not bind to any of the anchored BH3 domains tested. Nonetheless, when expressed inside cells the anchored PUMA and Bim BH3 α-helices powerfully induce cell death in the absence of efficient targeting to the mitochondrial membrane, whereas the Noxa helix requires a membrane insertion domain in order to kill Mcl-1-dependent myeloma cells. Finally, the binding of the Bim BH3 peptide to Bax was the only interaction with a pro-apoptotic effector protein observed in this study.

  11. Vibration Characteristic of Anchoring System of Bolt and Elastic Wave Propagation Law

    Institute of Scientific and Technical Information of China (English)

    WANG Cheng; LI Yi; NING Jian-guo; LIN Hua-chang

    2006-01-01

    Based on one-dimension wave theory, the propagation law of elastic wave along the rock bolt, rock medium and their coupling system are researched, and the attenuation law and propagation mechanism of wave in the anchoring system are obtained. Meanwhile, the studies on end reflection and dynamic response under load are also carried out experimentally, the relationship between anchoring length and excited wave length is obtained when the end reflection of bolt emerges, and it is concluded that under the condition of bolt loaded,as the load increases, the reflection of the upper interface of anchoring segment weakens while the end reflection strengthens relatively, hence the energy attenuation increases. These results provide some important theory basis for measuring the effective anchoring length of bolt, judging the bonding quality of anchoring end and surrounding rock, and estimating the utmost load force of bolt.

  12. Functionalized Nanostructures: Redox-Active Porphyrin Anchors for Supramolecular DNA Assemblies

    KAUST Repository

    Börjesson, Karl

    2010-09-28

    We have synthesized and studied a supramolecular system comprising a 39-mer DNA with porphyrin-modified thymidine nucleosides anchored to the surface of large unilamellar vesicles (liposomes). Liposome porphyrin binding characteristics, such as orientation, strength, homogeneity, and binding site size, was determined, suggesting that the porphyrin is well suited as a photophysical and redox-active lipid anchor, in comparison to the inert cholesterol anchor commonly used today. Furthermore, the binding characteristics and hybridization capabilities were studied as a function of anchor size and number of anchoring points, properties that are of importance for our future plans to use the addressability of these redox-active nodes in larger DNA-based nanoconstructs. Electron transfer from photoexcited porphyrin to a lipophilic benzoquinone residing in the lipid membrane was characterized by steady-state and time-resolved fluorescence and verified by femtosecond transient absorption. © 2010 American Chemical Society.

  13. The portion size effect on food intake. An anchoring and adjustment process?

    Science.gov (United States)

    Marchiori, David; Papies, Esther K; Klein, Olivier

    2014-10-01

    People consistently over-eat when served a large compared with a small (appropriate) portion of food. However, the mechanism underlying this so-called portion size effect is not well understood. We argue that the process of anchoring and adjustment naturally describes this effect, such that the size of a presented portion works as an anchor that strongly influences consumption. The classical anchoring and adjustment paradigm was applied to six hypothetical eating situations. Participants were asked to imagine being served either a small or a large portion of food (i.e., low and high anchor) and to indicate whether they would consume more or less than this amount. Then, they indicated how much they would eat. These estimates were compared with a no-anchor condition where participants did not imagine a specific portion size but only indicated how much they would eat. In addition, half of participants in the anchoring conditions received a discounting instruction stating that the portion size they had been asked to imagine was randomly selected and thus not informative for their consumption estimate. As expected, participants who imagined to be served larger portions estimated to consume significantly more food than participants in the no-anchor condition, and participants who imagined to be served smaller portions estimated to consume significantly less food than participants in the no-anchor condition. The discounting manipulation did not reduce this effect of the anchors. We suggest that the process of anchoring and adjustment may provide a useful framework to understand the portion size effect and we discuss implications of this perspective.

  14. Convenient synthesis and application of versatile nucleic acid lipid membrane anchors in the assembly and fusion of liposomes

    DEFF Research Database (Denmark)

    Ries, Oliver; Löffler, Philipp M. G.; Vogel, Stefan

    2015-01-01

    Hydrophobic moieties like lipid membrane anchors are highly demanded modifications for nucleic acid oligomers. Membrane-anchor modified oligonucleotides are applicable in biomedicine leading to new delivery strategies as well as in biophysical investigations towards assembly and fusion of liposom...

  15. Experimental Study On Lateral Load Capacity of Bamboo RC Beam Column Joints Strengthened By Bamboo Mechanical Anchors

    Directory of Open Access Journals (Sweden)

    Sri Umniati B.

    2017-01-01

    Full Text Available In this paper, the prospective of bamboos which available abundantly especially in Indonesia as rebars and mechanical anchors are studied. And also the endurance of the bamboos mechanical anchors to withstand cyclic loading were observed. Nine classes of bamboos bar were evaluated: consist of 3 different anchors (0, 4 and 8 anchors and 3 different compressive strength (19.19 MPa, 29.61 MPa and 37.96 MPa means 3 × 3 parameters. The results show that the lateral load capacity increased significantly with the present of bamboo anchors specimens: 26.04 % for 4 anchors specimens (C2 and 25 % for the 8 anchors specimens (C3 compared to zero anchor specimens (C1. On the other hand, the compressive strength of concrete have no significant effects to the lateral load capacity. Overall it can be concluded that, bamboo can be used as mechanical anchorage to strengthen beam column joint.

  16. Quantitative predictions of peptide binding to MHC class I molecules using specificity matrices and anchor-stratified calibrations

    DEFF Research Database (Denmark)

    Lauemøller, S L; Holm, A; Hilden, J;

    2001-01-01

    predictions, we have measured the MHC class I binding of a large number of peptides. In an attempt to further improve predictions and to include sequence dependency, we subdivided the panel of peptides according to whether the peptides had zero, one or two primary anchor residues. This allowed us to define......Peptides are key immune targets. They are generated by fragmentation of antigenic proteins, selected by major histocompatibility complex (MHC) molecules and subsequently presented to T cells. One of the most selective requirements is that of peptide binding to MHC. Accurate descriptions...... and predictions of peptide-MHC interactions are therefore important. Quantitative matrices representing MHC class I specificity can be used to search any query protein for the presence of MHC binding peptides. Assuming that each peptide residue contributes to binding in an additive and sequence independent manner...

  17. Activation of duck RIG-I by TRIM25 is independent of anchored ubiquitin.

    Directory of Open Access Journals (Sweden)

    Domingo Miranzo-Navarro

    Full Text Available Retinoic acid inducible gene I (RIG-I is a viral RNA sensor crucial in defense against several viruses including measles, influenza A and hepatitis C. RIG-I activates type-I interferon signalling through the adaptor for mitochondrial antiviral signaling (MAVS. The E3 ubiquitin ligase, tripartite motif containing protein 25 (TRIM25, activates human RIG-I through generation of anchored K63-linked polyubiquitin chains attached to lysine 172, or alternatively, through the generation of unanchored K63-linked polyubiquitin chains that interact non-covalently with RIG-I CARD domains. Previously, we identified RIG-I of ducks, of interest because ducks are the host and natural reservoir of influenza viruses, and showed it initiates innate immune signaling leading to production of interferon-beta (IFN-β. We noted that K172 is not conserved in RIG-I of ducks and other avian species, or mouse. Because K172 is important for both mechanisms of activation of human RIG-I, we investigated whether duck RIG-I was activated by TRIM25, and if other residues were the sites for attachment of ubiquitin. Here we show duck RIG-I CARD domains are ubiquitinated for activation, and ubiquitination depends on interaction with TRIM25, as a splice variant that cannot interact with TRIM25 is not ubiquitinated, and cannot be activated. We expressed GST-fusion proteins of duck CARD domains and characterized TRIM25 modifications of CARD domains by mass spectrometry. We identified two sites that are ubiquitinated in duck CARD domains, K167 and K193, and detected K63 linked polyubiquitin chains. Site directed mutagenesis of each site alone, does not alter the ubiquitination profile of the duck CARD domains. However, mutation of both sites resulted in loss of all attached ubiquitin and polyubiquitin chains. Remarkably, the double mutant duck RIG-I CARD still interacts with TRIM25, and can still be activated. Our results demonstrate that anchored ubiquitin chains are not necessary for

  18. Role of exchange protein directly activated by cAMP (EPAC1) in breast cancer cell migration and apoptosis.

    Science.gov (United States)

    Kumar, Naveen; Gupta, Sonal; Dabral, Surbhi; Singh, Shailja; Sehrawat, Seema

    2017-02-16

    Despite the current progress in cancer research and therapy, breast cancer remains the leading cause of mortality among half a million women worldwide. Migration and invasion of cancer cells are associated with prevalent tumor metastasis as well as high mortality. Extensive studies have powerfully established the role of prototypic second messenger cAMP and its two ubiquitously expressed intracellular cAMP receptors namely the classic protein kinaseA/cAMP-dependent protein kinase (PKA) and the more recently discovered exchange protein directly activated by cAMP/cAMP-regulated guanine nucleotide exchange factor (EPAC/cAMP-GEF) in cell migration, cell cycle regulation, and cell death. Herein, we performed the analysis of the Cancer Genome Atlas (TCGA) dataset to evaluate the essential role of cAMP molecular network in breast cancer. We report that EPAC1, PKA, and AKAP9 along with other molecular partners are amplified in breast cancer patients, indicating the importance of this signaling network. To evaluate the functional role of few of these proteins, we used pharmacological modulators and analyzed their effect on cell migration and cell death in breast cancer cells. Hence, we report that inhibition of EPAC1 activity using pharmacological modulators leads to inhibition of cell migration and induces cell death. Additionally, we also observed that the inhibition of EPAC1 resulted in disruption of its association with the microtubule cytoskeleton and delocalization of AKAP9 from the centrosome as analyzed by in vitro imaging. Finally, this study suggests for the first time the mechanistic insights of mode of action of a primary cAMP-dependent sensor, Exchange protein activated by cAMP 1 (EPAC1), via its interaction with A-kinase anchoring protein 9 (AKAP9). This study provides a new cell signaling cAMP-EPAC1-AKAP9 direction to the development of additional biotherapeutics for breast cancer.

  19. The protease resistant surface (PRS) glycoconjugate from Trypanosoma congolense has an inositol-acylated glycosylphosphatidylinositol anchor, containing a significant proportion of myristate at the sn-2 position.

    Science.gov (United States)

    Greganova, Eva; Bütikofer, Peter; Acosta-Serrano, Alvaro

    2010-05-01

    In the tsetse fly, the surface of Trypanosoma congolense parasites is covered by a dense layer of glycosylphosphatidylinositol (GPI)-anchored molecules. These include EPGENGT procyclin and protease resistant surface molecule (PRS), as well as congolense epimastigote-specific protein, CESP, and glutamic acid- and alanine-rich protein (GARP). The GPI structures of EPGENGT and GARP have been partially elucidated, but very little is known about PRS. We now purified PRS and analyzed its GPI lipid structure and carbohydrate composition using mass spectrometry. We found that unlike EPGENGT and GARP, the GPI anchor of PRS is unusually composed of inositol-acylated diacyl-phosphatidylinositols, including species containing either myristic or oleic acid at the sn-2 position of the glycerol backbone. This is the first identification of a tri-acylated GPI anchor containing myristate in procyclic form trypanosomes. In addition, we found that PRS is highly rich in galactose and sialic acid residues, suggesting that it may represent a major acceptor of the parasite trans-sialidase.

  20. Measuring the Activity of Leucine-Rich Repeat Kinase 2: A Kinase Involved in Parkinson's Disease

    Science.gov (United States)

    Lee, Byoung Dae; Li, Xiaojie; Dawson, Ted M.; Dawson, Valina L.

    2015-01-01

    Mutations in the LRRK2 (Leucine-Rich Repeat Kinase 2) gene are the most common cause of autosomal dominant Parkinson's disease. LRRK2 has multiple functional domains including a kinase domain. The kinase activity of LRRK2 is implicated in the pathogenesis of Parkinson's disease. Developing an assay to understand the mechanisms of LRRK2 kinase activity is important for the development of pharmacologic and therapeutic applications. Here, we describe how to measure in vitro LRRK2 kinase activity and its inhibition. PMID:21960214

  1. Career Anchors and the Effects of Downsizing: Implications for Generations and Cultures at Work. A Preliminary Investigation.

    Science.gov (United States)

    Marshall, Verena; Bonner, Dede

    2003-01-01

    The relationships among career anchors, age, culture, gender, employment experience, and the impact of career planning on downsizing were examined with data from 423 management students (49% had been downsized). Lifestyle was the most valued anchor across age groups, stability/security the least; compared with Schein's earlier anchors research,…

  2. The Place of White in a World of Grays: A Double-Anchoring Theory of Lightness Perception

    Science.gov (United States)

    Bressan, Paola

    2006-01-01

    The specific gray shades in a visual scene can be derived from relative luminance values only when an anchoring rule is followed. The double-anchoring theory I propose in this article, as a development of the anchoring theory of Gilchrist et al. (1999), assumes that any given region (a) belongs to one or more frameworks, created by Gestalt…

  3. Theoretical analysis of a new segmented anchoring style in weakly cemented soft surrounding rock

    Institute of Scientific and Technical Information of China (English)

    Zhao Zenghui; Wang Weiming; Wang Lihua

    2016-01-01

    According to the tensile failure of rock bolt in weakly cemented soft rock, this paper presents a new seg-mented anchoring style in order to weaken the cumulative effect of anchoring force associated with the large deformation. Firstly, a segmented mechanical model was established in which free and anchoring section of rock bolt were respectively arranged in different deformation zones. Then, stress and displace-ment in elastic non-anchoring zone, elastic anchoring zone, elastic sticking zone, softening sticking zone and broken zone were derived respectively based on neural theory and tri-linear strain softening constitutive model of soft rock. Results show that the anchoring effect can be characterized by a supporting parameter β. With its increase, the peak value of tangential stress gradually moves to the roadway wall, and the radial stress significantly increases, which means the decrease of equivalent plastic zone and improvement of confining effect provided by anchorage body. When β increases to 0.72, the equivalent plastic zone disappears, and stresses tend to be the elastic solutions. In addition, the anchoring effect on the displacement of surrounding rock can be quantified by a normalization factorδ.

  4. Modified method for external attachment of transmitters to birds using two subcutaneous anchors

    Science.gov (United States)

    Lewis, T.L.; Flint, P.L.

    2008-01-01

    Of the transmitter attachment techniques for birds, the subcutaneous anchor provides a secure attachment that yields relatively few secondary effects. However, the use of subcutaneous anchors has been limited by transmitter size and retention time. Using a modified method of attachment that utilized two subcutaneous anchors, we deployed 69 GPS transmitters, plus 13 VHF transmitters that were similar in size and weight to GPS models, on Pacific Black Brant (Branta bernicla nigricans). Prior to our study, only harnesses were used for attaching GPS transmitters on birds, mainly because GPS transmitters are too large for other external attachment techniques and implantation in the body cavity attenuates the GPS signal. Thus, to increase the size capacity of anchor attachment and to avoid the well-documented negative effects of harnesses on behavior and survival, we added a second anchor at the transmitter's posterior end. The double-anchor attachment technique was quickly and easily accomplished in the field, requiring bird handling times of killed by hunters (3-6 mo after deployment) retained their GPS transmitters. For studies involving the use of relatively large transmitters, the double-anchor method appears to provide a viable alternative for external attachment. ?? 2008 Association of Field Ornithologists.

  5. Rock engineering design of post-tensioned anchors for dams e A review

    Institute of Scientific and Technical Information of China (English)

    E.T. Brown

    2015-01-01

    abstract High-capacity, post-tensioned anchors have found wide-spread use, originally in initial dam design and construction, and more recently in the strengthening and rehabilitation of concrete dams to meet modern design and safety standards. Despite the advances that have been made in rock mechanics and rock engineering during the last 80 years in which post-tensioned anchors have been used in dam en-gineering, some aspects of the rock engineering design of high-capacity rock anchors for dams have changed relatively little over the last 30 or 40 years. This applies, in particular, to the calculations usually carried out to establish the grouted embedment lengths required for deep, post-tensioned anchors. These calculations usually make simplified assumptions about the distribution and values of rockegrout interface shear strengths, the shape of the volume of rock likely to be involved in uplift failure under the influence of a system of post-tensioned anchors, and the mechanism of that failure. The resulting designs are generally conservative. It is concluded that these aspects of the rock engineering design of large, post-tensioned rock anchors for dams can be significantly improved by making greater use of modern, comprehensive, numerical analyses in conjunction with three-dimensional (3D) models of the rock mass structure, realistic rock and rock mass properties, and the results of prototype anchor tests in the rock mass concerned.

  6. Anchoring effects in the judgment of confidence: semantic or numeric priming?

    Science.gov (United States)

    Carroll, Steven R; Petrusic, William M; Leth-Steensen, Craig

    2009-02-01

    Over the last decade, researchers have debated whether anchoring effects are the result of semantic or numeric priming. The present study tested both hypotheses. In four experiments involving a sensory detection task, participants first made a relative confidence judgment by deciding whether they were more or less confident than an anchor value in the correctness of their decision. Subsequently, they expressed an absolute level of confidence. In two of these experiments, the relative confidence anchor values represented the midpoints between the absolute confidence scale values, which were either explicitly numeric or semantic, nonnumeric representations of magnitude. In two other experiments, the anchor values were drawn from a scale modally different from that used to express the absolute confidence (i.e., nonnumeric and numeric, respectively, or vice versa). Regardless of the nature of the anchors, the mean confidence ratings revealed anchoring effects only when the relative and absolute confidence values were drawn from identical scales. Together, the results of these four experiments limit the conditions under which both numeric and semantic priming would be expected to lead to anchoring effects.

  7. Aurora A kinase regulates proper spindle positioning in C. elegans and in human cells.

    Science.gov (United States)

    Kotak, Sachin; Afshar, Katayon; Busso, Coralie; Gönczy, Pierre

    2016-08-01

    Accurate spindle positioning is essential for error-free cell division. The one-cell Caenorhabditis elegans embryo has proven instrumental for dissecting mechanisms governing spindle positioning. Despite important progress, how the cortical forces that act on astral microtubules to properly position the spindle are modulated is incompletely understood. Here, we report that the PP6 phosphatase PPH-6 and its associated subunit SAPS-1, which positively regulate pulling forces acting on spindle poles, associate with the Aurora A kinase AIR-1 in C. elegans embryos. We show that acute inactivation of AIR-1 during mitosis results in excess pulling forces on astral microtubules. Furthermore, we uncover that AIR-1 acts downstream of PPH-6-SAPS-1 in modulating spindle positioning, and that PPH-6-SAPS-1 negatively regulates AIR-1 localization at the cell cortex. Moreover, we show that Aurora A and the PP6 phosphatase subunit PPP6C are also necessary for spindle positioning in human cells. There, Aurora A is needed for the cortical localization of NuMA and dynein during mitosis. Overall, our work demonstrates that Aurora A kinases and PP6 phosphatases have an ancient function in modulating spindle positioning, thus contributing to faithful cell division.

  8. New benzimidazoles and their antitumor effects with Aurora A kinase and KSP inhibitory activities.

    Science.gov (United States)

    Abd El-All, Amira S; Magd-El-Din, Asmaa A; Ragab, Fatma A F; ElHefnawi, Mahmoud; Abdalla, Mohamed M; Galal, Shadia A; El-Rashedy, Ahmed A

    2015-07-01

    A newly synthesized series of anticancer compounds comprising thiazolo[3,2-a]pyrimidine derivatives 6a-q bearing a benzimidazole moiety was produced via a one-pot reaction of N-(4-(1H-benzo[d]imidazol-2-yl)phenyl)-2-cyanoacetamide 5 with 2-aminothiazole and an appropriate aromatic aldehyde. Compound 7 was obtained via the reaction of 4-(1H-benzo[d]imidazol-2yl)benzenamide 1 with carbon disulphide and methyl iodide in the presence of concentrated aqueous solution of NaOH, then treated with o-phenylenediamine to give N-(4-1H-benzo[d]imidazol-2-yl)phenyl)-1H-benzo[d]imidazol-2-amine 8. The structures of the newly synthesized compounds were confirmed by analytical and spectroscopic measurements (IR, MS, and (1) H NMR). The synthesized products were screened and studied for their in vitro antitumor activity against three human cancer cell lines (namely colorectal cancer cell line HCT116, human liver cancer cell line HepG2, and human ovarian cancer cell line A2780) and their Aurora A kinase and KSP inhibitory activities. All newly synthesized compounds revealed marked results comparable with the standard drug CK0106023. The compounds 6e and 6k of the thiazolopyrimidine derivatives were the most active compounds when tested against the three cell lines in comparison with the standard drug CK0106023, and showed potent dual KSP and Aurora A kinase inhibition.

  9. Red electroluminescence of ruthenium sensitizer functionalized by sulfonate anchoring groups.

    Science.gov (United States)

    Shahroosvand, Hashem; Abbasi, Parisa; Mohajerani, Ezeddin; Janghouri, Mohammad

    2014-06-28

    We have synthesized five novel Ru(ii) phenanthroline complexes with an additional aryl sulfonate ligating substituent at the 5-position [Ru(L)(bpy)2](BF4)2 (1), [Ru(L)(bpy)(SCN)2] (2), [Ru(L)3](BF4)2 (3), [Ru(L)2(bpy)](BF4)2 (4) and [Ru(L)(BPhen)(SCN)2] (5) (where L = 6-one-[1,10]phenanthroline-5-ylamino)-3-hydroxynaphthalene 1-sulfonic, bpy = 2,2'-bipyridine, BPhen = 4,7-diphenyl-1,10-phenanthroline), as both photosensitizers for oxide semiconductor solar cells (DSSCs) and light emitting diodes (LEDs). The absorption and emission maxima of these complexes red shifted upon extending the conjugation of the phenanthroline ligand. Ru phenanthroline complexes exhibit broad metal to ligand charge transfer-centered electroluminescence (EL) with a maximum near 580 nm. Our results indicated that a particular structure (2) can be considered as both DSSC and OLED devices. The efficiency of the LED performance can be tuned by using a range of ligands. Device (2) has a luminance of 550 cd m(-2) and maximum efficiency of 0.9 cd A(-1) at 18 V, which are the highest values among the five devices. The turn-on voltage of this device is approximately 5 V. The role of auxiliary ligands in the photophysical properties of Ru complexes was investigated by DFT calculation. We have also studied photovoltaic properties of dye-sensitized nanocrystalline semiconductor solar cells based on Ru phenanthroline complexes and an iodine redox electrolyte. A solar energy to electricity conversion efficiency (η) of 0.67% was obtained for Ru complex (2) under standard AM 1.5 irradiation with a short-circuit photocurrent density (Jsc) of 2.46 mA cm(-2), an open-circuit photovoltage (Voc) of 0.6 V, and a fill factor (ff) of 40%, which are all among the highest values for ruthenium sulfonated anchoring groups reported so far. Monochromatic incident photon to current conversion efficiency was 23% at 475 nm. Photovoltaic studies clearly indicated dyes with two SCN substituents yielded a higher Jsc for the

  10. A conserved P-loop anchor limits the structural dynamics that mediate nucleotide dissociation in EF-Tu

    Science.gov (United States)

    Mercier, Evan; Girodat, Dylan; Wieden, Hans-Joachim

    2015-01-01

    The phosphate-binding loop (P-loop) is a conserved sequence motif found in mononucleotide-binding proteins. Little is known about the structural dynamics of this region and its contribution to the observed nucleotide binding properties. Understanding the underlying design principles is of great interest for biomolecular engineering applications. We have used rapid-kinetics measurements in vitro and molecular dynamics (MD) simulations in silico to investigate the relationship between GTP-binding properties and P-loop structural dynamics in the universally conserved Elongation Factor (EF) Tu. Analysis of wild type EF-Tu and variants with substitutions at positions in or adjacent to the P-loop revealed a correlation between P-loop flexibility and the entropy of activation for GTP dissociation. The same variants demonstrate more backbone flexibility in two N-terminal amino acids of the P-loop during force-induced EF-Tu.GTP dissociation in Steered Molecular Dynamics simulations. Amino acids Gly18 and His19 are involved in stabilizing the P-loop backbone via interactions with the adjacent helix C. We propose that these P-loop/helix C interactions function as a conserved P-loop anchoring module and identify the presence of P-loop anchors within several GTPases and ATPases suggesting their evolutionary conservation.

  11. A conserved P-loop anchor limits the structural dynamics that mediate nucleotide dissociation in EF-Tu

    Science.gov (United States)

    Mercier, Evan; Girodat, Dylan; Wieden, Hans-Joachim

    2015-01-01

    The phosphate-binding loop (P-loop) is a conserved sequence motif found in mononucleotide-binding proteins. Little is known about the structural dynamics of this region and its contribution to the observed nucleotide binding properties. Understanding the underlying design principles is of great interest for biomolecular engineering applications. We have used rapid-kinetics measurements in vitro and molecular dynamics (MD) simulations in silico to investigate the relationship between GTP-binding properties and P-loop structural dynamics in the universally conserved Elongation Factor (EF) Tu. Analysis of wild type EF-Tu and variants with substitutions at positions in or adjacent to the P-loop revealed a correlation between P-loop flexibility and the entropy of activation for GTP dissociation. The same variants demonstrate more backbone flexibility in two N-terminal amino acids of the P-loop during force-induced EF-Tu·GTP dissociation in Steered Molecular Dynamics simulations. Amino acids Gly18 and His19 are involved in stabilizing the P-loop backbone via interactions with the adjacent helix C. We propose that these P-loop/helix C interactions function as a conserved P-loop anchoring module and identify the presence of P-loop anchors within several GTPases and ATPases suggesting their evolutionary conservation. PMID:25566871

  12. The 2.5 mm PushLock suture anchor system versus a traditional suture anchor for ulnar collateral ligament injuries of the thumb: a biomechanical study.

    Science.gov (United States)

    Jarrett, C D; McGillivary, G R; Hutton, W C

    2010-02-01

    We compared the biomechanical strength of the 2.5 mm PushLock suture anchor with a traditional Bio-SutureTak suture anchor in repair of ulnar collateral ligament injuries. Iatrogenic ulnar collateral ligament injuries in 18 cadaveric thumbs were repaired and used to test for load to failure and cyclic loading. The average force required to generate a 2 mm gap was 7.7 N for the 2.5 mm PushLock and 6.3 N for the Bio-SutureTak (p = 0.04). The ultimate load to failure was 28.0 N for the 2.5 mm PushLock and 18.8 N for the Bio-SutureTak (p = 0.16). There were no statistical differences between the two suture anchors under cyclic loading. The 2.5 mm PushLock suture anchor provides significantly stronger resistance to 2 mm gap formation at the repair site and is less likely to fail at the suture-ligament interface. However, there was no difference in the load to failure between the two suture anchors.

  13. Behavior of pre-stress group anchors--Theory approach and model

    Institute of Scientific and Technical Information of China (English)

    HE Siming; WANG Chenghua; QIAO Jianping

    2003-01-01

    First of all the pre-stress group anchor ropes are resolved into two sub-systems: the stable rock stand lateral resistance load and inner bonding section stand lateral resistance load and pre-stress load. Then, discretization of every sub-system was carried on and it is assumed that different micro-sections possess uniform distribution side resistance. On the basis of Mindlin stress solution, stress overlay principle, modified layered-summation method as well as the load transfer method, we study the anchor group effect and present a theory model which calculates the anchor group effect and establishes the relevant iterate standard.

  14. Suture anchor repair of patellar tendon rupture after total knee arthroplasty.

    Science.gov (United States)

    Kamath, Atul F; Shah, Roshan P; Summers, Nathan; Israelite, Craig L

    2013-12-01

    Extensor mechanism disruption after total knee arthroplasty (TKA) is a complex problem that often requires surgical repair for functional deficits. We present a brief technical note on suture anchor fixation of a patellar tendon rupture after TKA. A surgical technique and literature review follows. Although suture anchor fixation is well described for tendinous repairs in other areas of orthopedic surgery, no study has discussed the use of suture anchors in patellar tendon repair after TKA. The technique must be evaluated in more patients with longer follow-up before adoption.

  15. Extrusion of bone anchor suture following flexor digitorum profundus tendon avulsion injury repair.

    LENUS (Irish Health Repository)

    Tiong, William H C

    2011-09-01

    Flexor digitorum profundus (FDP) zone I tendon avulsion injury is traditionally repaired with a pullout suture technique. More recently, bone anchor sutures have been used as a viable alternative and have largely replaced areas in hand surgery where pullout suture technique was once required. To date, there have been very few complications reported related to bone anchor suture use in FDP tendon reattachment to the bone. We report a very unusual case of extrusion of bone anchor through the nailbed, 6 years after zone I FDP tendon avulsion injury repair and a brief review of literature.

  16. Study of Vertical Seismic Response of Concrete Self-Anchored Suspension Bridges

    Institute of Scientific and Technical Information of China (English)

    刘春城; 石磊; 张哲

    2004-01-01

    Based on the variational prineiple of incomplete generalized potential energy with large deflection, the vertical nonlinear vibrational differential equation of self-anchored suspension bridge is presented by taking the effect of coupling of flexural and axial action into consideration. The linear vertical equation is obtained by omitting the nonlinear term, and the pseudo excitation method(PEM). Taking the self-anchored concrete suspension bridge over Lanqi Songhua river for an example, the expected peak responses of main beam, towers and cables are calculated. And the seismic spatial effects on vertical seismic response of self-anchored suspension bridges are discussed.

  17. Experimental study on the stability of plate anchors in clay under cyclic loading

    Directory of Open Access Journals (Sweden)

    Long Yu

    2015-03-01

    Full Text Available Although the bearing capacity of plate anchors in clay has been studied extensively, the results considering the effects of offshore cyclic loading are relatively rare. In the present study, 1 g model tests are carried out to investigate the effect of cyclic loading on the bearing capacity of plate anchors in clay. The ultimate pullout capacity of plate anchors in clay decreases as the accumulated plastic shear strain grows due to the strain-softening of clay under cyclic loading. The load–displacement curves of these tests are presented and the effects of overburden stress and cyclic loading amplitude on the strain-softening behavior are discussed.

  18. Structure of the Membrane Anchor of Pestivirus Glycoprotein Erns, a Long Tilted Amphipathic Helix

    Science.gov (United States)

    Aberle, Daniel; Muhle-Goll, Claudia; Bürck, Jochen; Wolf, Moritz; Reißer, Sabine; Luy, Burkhard; Wenzel, Wolfgang; Ulrich, Anne S.; Meyers, Gregor

    2014-01-01

    Erns is an essential virion glycoprotein with RNase activity that suppresses host cellular innate immune responses upon being partially secreted from the infected cells. Its unusual C-terminus plays multiple roles, as the amphiphilic helix acts as a membrane anchor, as a signal peptidase cleavage site, and as a retention/secretion signal. We analyzed the structure and membrane binding properties of this sequence to gain a better understanding of the underlying mechanisms. CD spectroscopy in different setups, as well as Monte Carlo and molecular dynamics simulations confirmed the helical folding and showed that the helix is accommodated in the amphiphilic region of the lipid bilayer with a slight tilt rather than lying parallel to the surface. This model was confirmed by NMR analyses that also identified a central stretch of 15 residues within the helix that is fully shielded from the aqueous layer, which is C-terminally followed by a putative hairpin structure. These findings explain the strong membrane binding of the protein and provide clues to establishing the Erns membrane contact, processing and secretion. PMID:24586172

  19. A multidomain hub anchors the chromosome segregation and chemotactic machinery to the bacterial pole.

    Science.gov (United States)

    Yamaichi, Yoshiharu; Bruckner, Raphael; Ringgaard, Simon; Möll, Andrea; Cameron, D Ewen; Briegel, Ariane; Jensen, Grant J; Davis, Brigid M; Waldor, Matthew K

    2012-10-15

    The cell poles constitute key subcellular domains that are often critical for motility, chemotaxis, and chromosome segregation in rod-shaped bacteria. However, in nearly all rods, the processes that underlie the formation, recognition, and perpetuation of the polar domains are largely unknown. Here, in Vibrio cholerae, we identified HubP (hub of the pole), a polar transmembrane protein conserved in all vibrios, that anchors three ParA-like ATPases to the cell poles and, through them, controls polar localization of the chromosome origin, the chemotactic machinery, and the flagellum. In the absence of HubP, oriCI is not targeted to the cell poles, chemotaxis is impaired, and a small but increased fraction of cells produces multiple, rather than single, flagella. Distinct cytoplasmic domains within HubP are required for polar targeting of the three ATPases, while a periplasmic portion of HubP is required for its localization. HubP partially relocalizes from the poles to the mid-cell prior to cell division, thereby enabling perpetuation of the polar domain in future daughter cells. Thus, a single polar hub is instrumental for establishing polar identity and organization.

  20. Microtubule anchoring by cortical actin bundles prevents streaming of the oocyte cytoplasm.

    Science.gov (United States)

    Wang, Ying; Riechmann, Veit

    2008-01-01

    The localisation of the determinants of the body axis during Drosophila oogenesis is dependent on the microtubule (MT) cytoskeleton. Mutations in the actin binding proteins Profilin, Cappuccino (Capu) and Spire result in premature streaming of the cytoplasm and a reorganisation of the oocyte MT network. As a consequence, the localisation of axis determinants is abolished in these mutants. It is unclear how actin regulates the organisation of the MTs, or what the spatial relationship between these two cytoskeletal elements is. Here, we report a careful analysis of the oocyte cytoskeleton. We identify thick actin bundles at the oocyte cortex, in which the minus ends of the MTs are embedded. Disruption of these bundles results in cortical release of the MT minus ends, and premature onset of cytoplasmic streaming. Thus, our data indicate that the actin bundles anchor the MTs minus ends at the oocyte cortex, and thereby prevent streaming of the cytoplasm. We further show that actin bundle formation requires Profilin but not Capu and Spire. Thus, our results support a model in which Profilin acts in actin bundle nucleation, while Capu and Spire link the bundles to MTs. Finally, our data indicate how cytoplasmic streaming contributes to the reorganisation of the MT cytoskeleton. We show that the release of the MT minus ends from the cortex occurs independently of streaming, while the formation of MT bundles is streaming dependent.

  1. SYP73 Anchors the ER to the Actin Cytoskeleton for Maintenance of ER Integrity and Streaming in Arabidopsis.

    Science.gov (United States)

    Cao, Pengfei; Renna, Luciana; Stefano, Giovanni; Brandizzi, Federica

    2016-12-05

    The endoplasmic reticulum (ER) is an essential organelle that spreads throughout the cytoplasm as one interconnected network of narrow tubules and dilated cisternae that enclose a single lumen. The ER network undergoes extensive remodeling, which critically depends on membrane-cytoskeleton interactions [1]. In plants, the ER is also highly mobile, and its streaming contributes significantly to the movement of other organelles [2, 3]. The remodeling and motility of the plant ER rely mainly on actin [4] and to a minor extent on microtubules [5]. Although a three-way interaction between the ER, cytosolic myosin-XI, and F-actin mediates the plant ER streaming [6], the mechanisms underlying stable interaction of the ER membrane with actin are unknown. Early electron microscopy studies suggested a direct attachment of the plant ER with actin filaments [7, 8], but it is plausible that yet-unknown proteins facilitate anchoring of the ER membrane with the cytoskeleton. We demonstrate here that SYP73, a member of the plant Syp7 subgroup of SNARE proteins [9] containing actin-binding domains, is a novel ER membrane-associated actin-binding protein. We show that overexpression of SYP73 causes a striking rearrangement of the ER over actin and that, similar to mutations of myosin-XI [4, 10, 11], loss of SYP73 reduces ER streaming and affects overall ER network morphology and plant growth. We propose a model for plant ER remodeling whereby the dynamic rearrangement and streaming of the ER network depend on the propelling action of myosin-XI over actin coupled with a SYP73-mediated bridging, which dynamically anchors the ER membrane with actin filaments.

  2. Gold nanoparticles assembled with dithiocarbamate-anchored molecular wires

    Science.gov (United States)

    Reeler, Nini E. A.; Lerstrup, Knud A.; Somerville, Walter; Speder, Jozsef; Petersen, Søren V.; Laursen, Bo W.; Arenz, Matthias; Qiu, Xiaohui; Vosch, Tom; Nørgaard, Kasper

    2015-10-01

    A protocol for the bottom-up self-assembly of nanogaps is developed through molecular linking of gold nanoparticles (AuNPs). Two π-conjugated oligo(phenylene ethynylene) molecules (OPE) with dithiocarbamate anchoring groups are used as ligands for the AuNPs. OPE-4S with a dithiocarbamate in each end of the molecule and a reference molecule OPE-2S with only a single dithiocarbamate end group. The linking mechanism of OPE-4S is investigated by using a combination of TEM, UV-Vis absorption and surface enhanced Raman spectroscopy (SERS) as well as studying the effect of varying the OPE-4S to AuNP concentration ratio. UV-Vis absorption confirms the formation of AuNP aggregates by the appearance of an extended plasmon band (EPB) for which the red shift and intensity depend on the OPE-4S:AuNP ratio. SERS confirms the presence of OPE-4S and shows a gradual increase of the signal intensity with increasing OPE-4S:AuNP ratios up to a ratio of about 4000, after which the SERS intensity does not increase significantly. For OPE-2S, no linking is observed below full coverage of the AuNPs indicating that the observed aggregate formation at high OPE-2S:AuNP ratios, above full AuNP coverage, is most likely of a physical nature (van der Waals forces or π-π interactions).

  3. High performance supercapacitors using metal oxide anchored graphene nanosheet electrodes

    KAUST Repository

    Baby, Rakhi Raghavan

    2011-01-01

    Metal oxide nanoparticles were chemically anchored onto graphene nanosheets (GNs) and the resultant composites - SnO2/GNs, MnO2/GNs and RuO2/GNs (58% of GNs loading) - coated over conductive carbon fabric substrates were successfully used as supercapacitor electrodes. The results showed that the incorporation of metal oxide nanoparticles improved the capacitive performance of GNs due to a combination of the effect of spacers and redox reactions. The specific capacitance values (with respect to the composite mass) obtained for SnO2/GNs (195 F g-1) and RuO 2/GNs (365 F g-1) composites at a scan rate of 20 mV s-1 in the present study are the best ones reported to date for a two electrode configuration. The resultant supercapacitors also exhibited high values for maximum energy (27.6, 33.1 and 50.6 W h kg-1) and power densities (15.9, 20.4 and 31.2 kW kg-1) for SnO2/GNs, MnO2/GNs and RuO2/GNs respectively. These findings demonstrate the importance and great potential of metal oxide/GNs based composite coated carbon fabric in the development of high-performance energy-storage systems. © 2011 The Royal Society of Chemistry.

  4. Emergence of Anchored Flux Tubes Through the Convection Zone

    CERN Document Server

    Fisher, George H; McClymont, Alexander N

    2010-01-01

    We model the evolution of buoyant magnetic flux tubes in the Sun's convection zone. A flux tube is assumed to lie initially near the top of the stably stratified radiative core below the convection zone, but a segment of it is perturbed into the convection zone by gradual heating and convective overshoot motions. The ends ("footpoints") of the segment remain anchored at the base of the convection zone, and if the segment is sufficiently long, it may be buoyantly unstable, rising through the convection zone in a short time. The length of the flux tube determines the ratio of buoyancy to magnetic tension: short loops of flux are arrested before reaching the top of the convection zone, while longer loops emerge to erupt through the photosphere. Using Spruit's convection zone model, we compute the minimum footpoint separation $L_c$ required for erupting flux tubes. We explore the dependence of $L_c$ on the initial thermal state of the perturbed flux tube segment and on its initial magnetic field strength. Followi...

  5. Dynamics of anchor last deployment of submersible buoy system

    Science.gov (United States)

    Zheng, Zhongqiang; Xu, Jianpeng; Huang, Peng; Wang, Lei; Yang, Xiaoguang; Chang, Zongyu

    2016-02-01

    Submersible buoy systems are widely used for oceanographic research, ocean engineering and coastal defense. Severe sea environment has obvious effects on the dynamics of submersible buoy systems. Huge tension can occur and may cause the snap of cables, especially during the deployment period. This paper studies the deployment dynamics of submersible buoy systems with numerical and experimental methods. By applying the lumped mass approach, a three-dimensional multi-body model of submersible buoy system is developed considering the hydrodynamic force, tension force and impact force between components of submersible buoy system and seabed. Numerical integration method is used to solve the differential equations. The simulation output includes tension force, trajectory, profile and dropping location and impact force of submersible buoys. In addition, the deployment experiment of a simplified submersible buoy model was carried out. The profile and different nodes' velocities of the submersible buoy are obtained. By comparing the results of the two methods, it is found that the numerical model well simulates the actual process and conditions of the experiment. The simulation results agree well with the results of the experiment such as gravity anchor's location and velocities of different nodes of the submersible buoy. The study results will help to understand the conditions of submersible buoy's deployment, operation and recovery, and can be used to guide the design and optimization of the system.

  6. Comparison of the tendon damage caused by four different anchor systems used in transtendon rotator cuff repair.

    Science.gov (United States)

    Zhang, Qing-Song; Liu, Sen; Zhang, Qiuyang; Xue, Yun; Ge, Dongxia; O'Brien, Michael J; Savoie, Felix H; You, Zongbing

    2012-01-01

    Objectives. The objective of this study was to compare the damage to the rotator cuff tendons caused by four different anchor systems. Methods. 20 cadaveric human shoulder joints were used for transtendon insertion of four anchor systems. The Healix Peek, Fastin RC, Bio-Corkscrew Suture, and Healix Transtend anchors were inserted through the tendons using standard transtendon procedures. The areas of tendon damage were measured. Results. The areas of tendon damage (mean ± standard deviation, n = 7) were 29.1 ± 4.3 mm(2) for the Healix Peek anchor, 20.4 ± 2.3 mm(2) for the Fastin RC anchor, 23.4 ± 1.2 mm(2) for the Bio-Corkscrew Suture anchor, 13.7 ± 3.2 mm(2) for the Healix Transtend anchor inserted directly, and 9.1 ± 2.1 mm(2) for the Healix Transtend anchor inserted through the Percannula system (P anchors). Conclusions. In a cadaver transtendon rotator cuff repair model, smaller anchors caused less damage to the tendon tissues. The Healix Transtend implant system caused the least damage to the tendon tissues. Our findings suggest that smaller anchors should be considered when performing transtendon procedures to repair partial rotator cuff tears.

  7. 锚链直径对船舶锚泊能力的影响%Influence of Diameter of Anchor Chains on the Anchoring Capability of Ships

    Institute of Scientific and Technical Information of China (English)

    胡晓芳; 丁德勇

    2014-01-01

    In order to meet the demand of anchor system using thinner anchor chains,this paper deduces the catenary equation by taking every anchor chain link as an infinitesimal. Based on the equation,the length of anchor chains and the maximum environmental forces and water depth are calculated for anchor chains with different diameters. According to the calculation result and actual practices,it is concluded that though the thicker the anchor chain,the better the anchoring capability,the applications of thinner chains are more reasonable when not using anchors with high holding power,or when the situation does not demand any special request of the mooring region or water depth.%从减轻船总体负担、采用链径更小的海洋系泊链的设计需求出发,以单个链环为微元对锚链在锚泊状态所形成悬链线方程进行推导,建立锚链在典型锚泊状态下的悬链线方程,并以此为基础完成不同链径锚链在抛锚长度、最大可承受环境力、最大抛锚深度等方面的计算分析。结合计算结果和船舶实际使用情况,对采用不同链径锚链的锚泊能力进行综合分析后认为,虽然链径较粗的锚链的理论锚泊能力较强,但若采用霍尔锚等非大抓力锚,或对锚泊水域面积无明确要求、没有在深水中抛锚的特殊需求等,则可以选用链径相对较细的海洋系泊链代替目前规范中规定的电焊锚链。

  8. Dynamic Effects of Anchor Positional Tolerance on Tension Moored Floating Wind Turbine

    Science.gov (United States)

    Wright, Christopher; Pakrashi, Vikram; Murphy, Jimmy

    2016-09-01

    For water depths greater than 60m floating wind turbines will become the most economical option for generating offshore wind energy. Tension mooring stabilised units are one type of platform being considered by the offshore wind energy industry. The complex mooring arrangement used by this type of platform means that the dynamics are greatly effected by offsets in the positioning of the anchors. This paper examines the issue of tendon anchor position tolerances. The dynamic effects of three positional tolerances are analysed in survival state using the time domain FASTLink. The severe impact of worst case anchor positional offsets on platform and turbine survivability is shown. The worst anchor misposition combinations are highlighted and should be strongly avoided. Novel methods to mitigate this issue are presented.

  9. Factors affecting the dynamic response of pre-stressed anchors after transient excitation

    Institute of Scientific and Technical Information of China (English)

    Xu Huijun; Li Qingfeng

    2011-01-01

    The wide application of pre-stressed bolting technology in coal mine tunnels has made the nondestructive stress wave reflection method of determining bolting quality an important one.The effect of the support plate on the dynamic response of the pre-stressed anchor is of particular interest.A theoreticalanalysis and numerical simulations are used to identify the factors affecting the contact stress between the support plate and the rock wall.A formula allowing the calculation of contact stress is presented.Stress wave propagation through the nut,support plate,and rock wall are predicted.The dynamic response signals were measured in the field using prestressed anchors pre-tightened to different torques.The effects from the support plate on the dynamic response were recorded and the results compared to the predictions of pre-stressed anchor.This work provides a theoretical reference for the signal processing of dynamic reflected wave signals in anchor bolts.

  10. Stone anchors from Bet Dwarka Island, Gujarat, Coast, India: Significance to historical period maritime activities

    Digital Repository Service at National Institute of Oceanography (India)

    Sundaresh; Gaur, A.S.; Tripati, S.; Gudigar, P.; Bandodkar, S.N.

    Bet Dwarka Island is situated on the extreme west of Indian territory in Jamnagar district of Gujarat. Underwater, the most preserved remains of ancient maritime activity could be the stone anchors of different types, as every boat requires...

  11. Optimal design of anchor cables for slope reinforcement based on stress and displacement fields

    Institute of Scientific and Technical Information of China (English)

    Guanghua Yang; Zhihui Zhong; Yucheng Zhang; Xudong Fu

    2015-01-01

    How to determine reasonable position and length of anchor cable is a frequently encountered but not well addressed problem in slope reinforcement projects. In this paper, the variable-modulus elastoplastic strength reduction method (SRM) is used to obtain the stress field, displacement field, and factor of safety of slope. Slope reinforcement using anchor cables is modeled by surface loading, i.e. different distributions of surface loading represent various reinforcement schemes. Optimal reinforcement scheme of anchor cables can be determined based on slope stress and displacement fields. By comparing the factor of safety and stress field before and after slope reinforcement, it is found that better rein-forcement results can be achieved if strong reinforcement is applied upon the regions with high stress and large displacement. This method can well optimize the arrangement of anchor cables. In addition, several cases are employed to verify the proposed method.

  12. 78 FR 45104 - Model Manufactured Home Installation Standards: Ground Anchor Installations

    Science.gov (United States)

    2013-07-26

    ... positioning of the test rig supports and their proximity to the anchor assembly being tested (Sec. .3285.402(b... examine labor market, or revealed preference, studies. Using the DOT estimate, avoiding one death in...

  13. On Inferring Demand for Health Care in the Presence of Anchoring, Acquiescence, and Selection Biases

    OpenAIRE

    Jay Bhattacharya; Adam Isen

    2008-01-01

    In the contingent valuation literature, both anchoring and acquiescence biases pose problems when using an iterative bidding game to infer willingness to pay. Anchoring bias occurs when the willingness to pay estimate is sensitive to the initially presented starting value. Acquiescence bias occurs when survey respondents exhibit a tendency to answer 'yes' to questions, regardless of their true preferences. More generally, whenever a survey format is used and not all of those contacted partici...

  14. Prediction of signal peptides and signal anchors by a hidden Markovmodel

    DEFF Research Database (Denmark)

    Nielsen, Henrik; Krogh, Anders Stærmose

    1998-01-01

    A hidden Markov model of signal peptides has been developed. It contains submodels for the N-terminal part, the hydrophobic region and the region around the cleavage site. For known signal peptides, the model can be used to assign objective boundaries between these three regions. Applied to our d...... is the poor discrimination between signal peptides and uncleaved signal anchors, but this is substantially improved by the hidden Markov model when expanding it with a very simple signal anchor model....

  15. Uplift of Symmetrical Anchor Plates by Using Grid-Fixed Reinforced Reinforcement in Cohesionless Soil

    Institute of Scientific and Technical Information of China (English)

    Hamed Niroumand; Khairul Anuar Kassim

    2014-01-01

    Uplift response of symmetrical anchor plates with and without grid fixed reinforced (GFR) reinforcement was evaluated in model tests and numerical simulations by Plaxis. Many variations of reinforcement layers were used to reinforce the sandy soil over symmetrical anchor plates. In the current research, different factors such as relative density of sand, embedment ratios, and various GFR parameters including size, number of layers, and the proximity of the layer to the symmetrical anchor plate were investigated in a scale model. The failure mechanism and the associated rupture surface were observed and evaluated. GFR, a tied up system made of fiber reinforcement polymer (FRP) strips and end balls, was connected to the geosynthetic material and anchored into the soil. Test results showed that using GFR reinforcement significantly improved the uplift capacity of anchor plates. It was found that the inclusion of one layer of GFR, which rested directly on the top of the anchor plate, was more effective in enhancing the anchor capacity itself than other methods. It was found that by including GFR the uplift response was improved by 29%. Multi layers of GFR proved more effective in enhancing the uplift capacity than a single GFR reinforcement. This is due to the additional anchorage provided by the GFR at each level of reinforcement. In general, the results show that the uplift capacity of symmetrical anchor plates in loose and dense sand can be significantly increased by the inclusion of GFR. It was also observed that the inclusion of GFR reduced the requirement for a large L/D ratio to achieve the required uplift capacity. The laboratory and numerical analysis results are found to be in agreement in terms of breakout factor and failure mechanism pattern.

  16. Why Do Countries Peg the Way They Peg? The Determinants of Anchor Currency Choice

    OpenAIRE

    Christopher M. Meissner; Oomes, Nienke

    2006-01-01

    Conditional on choosing a pegged exchange rate regime, what determines the currency to which countries peg or “anchor” their exchange rate? This paper aims to answer this question using a panel multinomial logit framework, covering more than 100 countries for the period 1980-1998. We find that trade network externalities are a key determinant of anchor currency choice, implying that there are multiple steady states for the distribution of anchor currencies in the international monetary system...

  17. Prediction of signal peptides and signal anchors by a hidden Markov model

    DEFF Research Database (Denmark)

    Krogh, Anders Stærmose; Nielsen, Henrik

    1998-01-01

    A hidden Markov model of signal peptides has been developed. It contains submodels for the N-terminal part, the hydrophobic region, and the region around the cleavage site. For known signal peptides, the model can be used to assign objective boundaries between these three regions. Applied to our ...... is the poor discrimination between signal peptides and uncleaved signal anchors, but this is substantially improved by the hidden Markov model when expanding it with a very simple signal anchor model....

  18. Training troubleshooting skills with an anchored instruction module in an authentic computer based simulation environment

    OpenAIRE

    2013-01-01

    To improve the application and transfer of troubleshooting skills when diagnosing faults in complex automated production units, we developed and implemented an “anchored instruction” learning module in the context of a computer based simulation environment. The effects of the instructional module were evaluated in a quasi-experimental evaluation study. During the study 42 mechatronic apprentices were trained in two parallel experimental groups with and without the anchored instruction module....

  19. Psychological career resources as predictors of working adults’ career anchors: An exploratory study

    OpenAIRE

    Melinde Coetzee; Dries Schreuder

    2009-01-01

    This study explored the relationship between the psychological career resources and career anchors of a sample of 2 997 working adults at predominantly managerial and supervisory levels in the service industry. The Psychological Career Resources Inventory and the Career Orientations Inventory were applied. Stepwise regression analyses indicated dimensions of psychological career resources as significant predictors of  participants’ career anchors. The findings add valuable new knowledge that ...

  20. Dynamic behavior of anchors in cracked and uncracked concrete: A progress report

    Energy Technology Data Exchange (ETDEWEB)

    Rodriguez, M.; Yong-gang Zhang; Lotze, D. [Univ. of Texas, Austin, TX (United States)] [and others

    1995-04-01

    In early 1993, the US Nuclear Regulatory Commission began a research program at The University of Texas at Austin, dealing with the dynamic behavior of anchors in cracked and uncracked concrete. In this paper, the progress of that research program is reviewed. The test program is summarized, the work performed to date is reviewed, with emphasis on the dynamic and static behavior of single tensile anchors in uncracked concrete. General conclusions from that work are discussed, and future plans are presented.

  1. Installation of concrete expansion anchors at the Fast Flux Test Facility

    Energy Technology Data Exchange (ETDEWEB)

    Clark, G.L.

    1980-01-01

    Installation criteria utilized at the Fast Flux Test Facility for concrete expansion anchors are presented. Static and dynamic load capabilities of various anchor types are discussed in relation to design loads, with particular emphasis placed on the yield load (the proportional limit). Effects of several variables (i.e., installation torque, hole diameter) are also investigated. Resolution and documentation of field problems (e.g., improper spacing, embedment, angularity) are also described. Recommendations for improving and controlling future installations are given.

  2. An exploratory study on the career anchors of educators in Singapore.

    Science.gov (United States)

    Tan, H H; Quek, B C

    2001-09-01

    In this exploratory study, the authors adopted an empirical approach to determine the various primary career anchors possessed by Singapore educators, the impact of the degree of congruency between teaching and the career anchors on intrinsic and extrinsic satisfaction, and turnover intentions. A number of hypotheses were supported and provided the 1st step in understanding this group of important individuals who nuture the future generation of human capital for the workplace.

  3. Webtag: a new web tool providing tags/anchors for RT-PCR experiments with prokaryotes

    OpenAIRE

    Lindblad Peter; Svensson Håkan; Pinto Fernando

    2007-01-01

    Abstract Background Webtag is a tool providing oligonucleotide sequences (usually called tags or anchors) that are absent from a specified genome. These tags/anchors can be appended to gene specific primers for reverse transcriptase polymerase chain reaction experiments, circumventing genomic DNA contamination. Results The use of a relational database, in conjunction with a series of scripts written in PHP and Perl, allows the user to rapidly obtain tags that are: 1) suitable for a specific o...

  4. Dynamic Active Earth Pressures of the Retaining Piles with Anchors under Vehicle Loads

    OpenAIRE

    Hong-zhi Qiu; Ji-ming Kong; Ren-chao Wang

    2016-01-01

    The pile-anchor supporting structure is widely used in foundation pit engineering; then knowledge of active earth pressure on piles is very important for engineers. In this paper, based on the pseudodynamic method and considering the vehicle’s vibration characteristic, a method to calculate the earth pressure on piles under vehicle load is presented. At the same time, the constraint of anchor is simplified relation of lateral deformation of piles in present method. Effects of a wide range of ...

  5. Discriminative ability of two different external anchors: improvement appears better than importance

    DEFF Research Database (Denmark)

    Lauridsen, Henrik Hein

    2009-01-01

    Title:    Discriminative ability of two different external anchors: improvement appears better than importance. Authors and affiliation: Henrik H. Lauridsen, DC, MSc, PhD*, Claus Manniche, MD, DMSc‡, Werner Vach, PhD §, Niels Grunnet-Nilsson DC, MD, PhD*, Jan Hartvigsen, DC, PhD*† * Clinical...... better discriminative abilities compared to the importance criterion. Consequently, we recommend using improvement and not importance as an anchor to determine the MIC....

  6. Discriminative ability of two different external anchors: improvement appears better than importance

    DEFF Research Database (Denmark)

    Lauridsen, Henrik Hein

    Title:    Discriminative ability of two different external anchors: improvement appears better than importance. Authors and affiliation: Henrik H. Lauridsen, DC, MSc, PhD*, Claus Manniche, MD, DMSc‡, Werner Vach, PhD §, Niels Grunnet-Nilsson DC, MD, PhD*, Jan Hartvigsen, DC, PhD*†   * Clinical...... better discriminative abilities compared to the importance criterion. Consequently, we recommend using improvement and not importance as an anchor to determine the MIC....

  7. Effects of surface anchoring on the electric Frederiks transition in ferronematic systems

    Science.gov (United States)

    Farrokhbin, Mojtaba; Kadivar, Erfan

    2016-11-01

    The effects of anchoring phenomenon on the electric Frederiks transition threshold field in a nematic liquid crystal doped with ferroelectric nanoparticles are discussed. The polarizability of these nanoparticles in combination with confinement effects cause the drastic effects on the ferronematic systems. This study is based on Frank free energy and Rapini-Papoular surface energy for ferronematic liquid crystal having finite anchoring condition. In the case of different anchoring boundary conditions, the Euler-Lagrange equation of the total free energy is numerically solved by using the finite difference method together with the relaxation method and Maxwell construction to select the physical solutions and therefore investigate the effects of different anchoring strengths on the Frederiks transition threshold field. Maxwell construction method is employed to select three periodic solutions for nematic liquid crystal director at the interfaces of a slab. In the interval from zero to half- π, there is only one solution for the director orientation. In this way, NLC director rotates toward the normal to the surface as the applied electric field increases at the walls. Our numerical results illustrate that above Frederiks transition and in the intermediate anchoring strength, nematic molecules illustrate the different orientation at slab boundaries. We also study the effects of different anchoring strengths, nanoparticle volume fractions and polarizations on the Frederiks transition threshold field. We report that decreasing in the nanoparticle polarization results in the saturation Frederiks threshold. However, this situation does not happen for the nanoparticles volume fraction.

  8. Study on interaction between soil and anchor chain with finite element method

    Science.gov (United States)

    Li, Sa; Xu, Bao-zhao; Wu, Yun-zhou; Li, Zhong-gang

    2016-12-01

    With the development of offshore engineering, deeply embedded anchors are needed to be penetrated to appreciable depth and attached at the pad-eye. The interaction between anchor chain and soil is a very complex process and has not been thoroughly understood yet. In this paper, the finite element method (FEM) was used to study the interaction of soil-chain system. Results of the analysis show that when the attachment point is at a shallow depth, the load-development characteristics of the chain from FEM are in good agreement with that from the model tests and theoretical analysis. But with the depth increment, the results are different obviously in different methods. This phenomenon is resulted from a variety of reasons, and the plastic zone around the chain was studied to try finding the mechanism behind it. It could be seen that the plastic zone extended in different modes at different depths of attachment points. The interaction between the soil and anchor chain makes the load acting on the anchor decrease, but the soil disturbed surrounding the chain increases the anchor failure possibility. When the anchor bearing capacity is evaluated, these two factors should be considered properly at the same time.

  9. Effects of GPI-anchored TNAP on the dynamic structure of model membranes

    Science.gov (United States)

    Garcia, A. F.; Simão, A. M. S.; Bolean, M; Hoylaerts, M. F.; Millán, J. L.; Ciancaglini, P; Costa-Filho, A. J.

    2017-01-01

    Tissue-nonspecific alkaline phosphatase (TNAP) plays a crucial role during skeletal mineralization, and TNAP deficiency leads to the soft bone disease hypophosphatasia. TNAP is anchored to the external surface of the plasma membranes by means of a GPI (glycosylphosphatidylinositol) anchor. Membrane-anchored and solubilized TNAP displays different kinetic properties against physiological substrates, indicating that membrane anchoring influences the enzyme function. Here, we used Electron Spin Resonance (ESR) measurements along with spin labeled phospholipids to probe the possible dynamic changes prompted by the interaction of GPI-anchored TNAP with model membranes. The goal was to systematically analyze the ESR data in terms of line shape changes and of alterations in parameters such as rotational diffusion rates and order parameters obtained from non-linear least-squares simulations of the ESR spectra of probes incorporated into DPPC liposomes and proteoliposomes. Overall, the presence of TNAP increased the dynamics and decreased the ordering in the three distinct regions probed by the spin labeled lipids DOPTC (headgroup), and 5- and 16-PCSL (acyl chains). The largest change was observed for 16-PCSL, thus suggesting that GPI-anchored TNAP can give rise to long reaching modifications that could influence membrane processes halfway through the bilayer. PMID:26389140

  10. Design and Application of a Field Sensing System for Ground Anchors in Slopes

    Directory of Open Access Journals (Sweden)

    Hyo Seon Park

    2013-03-01

    Full Text Available In a ground anchor system, cables or tendons connected to a bearing plate are used for stabilization of slopes. Then, the stability of a slope is dependent on maintaining the tension levels in the cables. So far, no research on a strain-based field sensing system for ground anchors has been reported. Therefore, in this study, a practical monitoring system for long-term sensing of tension levels in tendons for anchor-reinforced slopes is proposed. The system for anchor-reinforced slopes is composed of: (1 load cells based on vibrating wire strain gauges (VWSGs, (2 wireless sensor nodes which receive and process the signals from load cells and then transmit the result to a master node through local area communication, (3 master nodes which transmit the data sent from sensor nodes to the server through mobile communication, and (4 a server located at the base station. The system was applied to field sensing of ground anchors in the 62 m-long and 26 m-high slope at the side of the highway. Based on the long-term monitoring, the safety of the anchor-reinforced slope can be secured by the timely applications of re-tensioning processes in tendons.

  11. Liquid-crystal anchoring transitions on aligning substrates processed by a plasma beam.

    Science.gov (United States)

    Yaroshchuk, Oleg V; Kiselev, Alexei D; Kravchuk, Ruslan M

    2008-03-01

    We have studied a sequence of anchoring transitions observed in nematic liquid crystals (NLCs) sandwiched between hydrophobic polyimide substrates treated with a plasma beam. There is a pronounced continuous transition from a homeotropic to a slightly tilted (nearly planar) alignment with the easy axis parallel to the incidence plane of the plasma beam (the zenithal transition) which takes place as the exposure dose increases. In NLCs with positive dielectric anisotropy, a further increase in the exposure dose results in in-plane reorientation of the easy axis by 90 degrees (the azimuthal transition). This transition occurs through the twofold degenerate alignment characteristic of second-order anchoring transitions. In contrast to the critical behavior of anchoring, the contact angle of the NLC and water on the treated substrates declines monotonically with increasing exposure dose. It follows that the surface concentration of hydrophobic chains decreases continuously. The anchoring transitions under consideration are qualitatively interpreted by using a simple phenomenological model of competing easy axes which is studied by analyzing anchoring diagrams of generalized polar and nonpolar anchoring models.

  12. Fragile X gene instability: Anchoring AGGs and linked microsatellites

    Energy Technology Data Exchange (ETDEWEB)

    Zhong, Nan; Yang, Weihong; Dobkin, C. [Institute for Basic Research in Developmental Disabilities, Staten Island, NY (United States)] [and others

    1995-08-01

    Interspersed AGGs within the FMR1 gene CGG repeat region may anchor the sequence and prevent slippage during replication. In order to detect the AGG position variations, we developed a method employing partial MnlI restriction analysis and analyzed X chromosomes from 187 males, including 133 normal controls (17 with 20-34 and 16 with 35-52 repeats), plus 54 fragile X premutations with 56-180 repeats. Among controls, the interspersed AGG positions were highly polymorphic, with heterozygosity of 91%. Among the control samples, 1.5% had no AGG positions, 25% had one, 71% had two, and 3% had three. Among the fragile X premutation samples, 63% had no AGG, while 37% had only one AGG. Analysis of premutation samples within fragile X families showed that variation occurred only within the 3{prime} end of the region. Thus, the instability was polar. Controls with {ge}15 pure CGG repeats were associated with the longest alleles of two nearby microsatellites, FRAXAC1 with 20-21 repeats and DXS548 with 202-206 bp and with increased microsatellite heterzygosity. The association of long pure CGG regions, as with fragile X chromosomes, with the longer and more heterozygous microsatellite alleles suggests they may be related mechanistically. Further, our results do not support a recent suggestion that the frequency of fragile X alleles may be increasing. Finally, analysis of a set of nonhuman primate samples showed that long pure CGG tracks are variable in size and are located within the 3{prime} region, which suggests that polar instability within FMR1 is evolutionarily quite old. 55 refs., 3 figs., 4 tabs.

  13. The late endosome/lysosome-anchored p18-mTORC1 pathway controls terminal maturation of lysosomes

    Energy Technology Data Exchange (ETDEWEB)

    Takahashi, Yusuke; Nada, Shigeyuki; Mori, Shunsuke; Soma-Nagae, Taeko; Oneyama, Chitose [Department of Oncogene Research, Research Institute for Microbial Diseases, Osaka University, 3-1 Yamadaoka, Suita, Osaka 565-0871 (Japan); Okada, Masato, E-mail: okadam@biken.osaka-u.ac.jp [Department of Oncogene Research, Research Institute for Microbial Diseases, Osaka University, 3-1 Yamadaoka, Suita, Osaka 565-0871 (Japan)

    2012-01-27

    Highlights: Black-Right-Pointing-Pointer p18 is a membrane adaptor that anchors mTORC1 to late endosomes/lysosomes. Black-Right-Pointing-Pointer We examine the role of the p18-mTORC1 pathway in lysosome biogenesis. Black-Right-Pointing-Pointer The loss of p18 causes accumulation of intact late endosomes by arresting lysosome maturation. Black-Right-Pointing-Pointer Inhibition of mTORC1 activity with rapamycin phenocopies the defects of p18 loss. Black-Right-Pointing-Pointer The p18-mTORC1 pathway plays crucial roles in the terminal maturation of lysosomes. -- Abstract: The late endosome/lysosome membrane adaptor p18 (or LAMTOR1) serves as an anchor for the mammalian target of rapamycin complex 1 (mTORC1) and is required for its activation on lysosomes. The loss of p18 causes severe defects in cell growth as well as endosome dynamics, including membrane protein transport and lysosome biogenesis. However, the mechanisms underlying these effects on lysosome biogenesis remain unknown. Here, we show that the p18-mTORC1 pathway is crucial for terminal maturation of lysosomes. The loss of p18 causes aberrant intracellular distribution and abnormal sizes of late endosomes/lysosomes and an accumulation of late endosome specific components, including Rab7, RagC, and LAMP1; this suggests that intact late endosomes accumulate in the absence of p18. These defects are phenocopied by inhibiting mTORC1 activity with rapamycin. Loss of p18 also suppresses the integration of late endosomes and lysosomes, resulting in the defective degradation of tracer proteins. These results suggest that the p18-mTORC1 pathway plays crucial roles in the late stages of lysosomal maturation, potentially in late endosome-lysosome fusion, which is required for processing of various macromolecules.

  14. Assembly and activation of alternative complement components on endothelial cell-anchored ultra-large von Willebrand factor links complement and hemostasis-thrombosis.

    Directory of Open Access Journals (Sweden)

    Nancy A Turner

    Full Text Available BACKGROUND: Vascular endothelial cells (ECs express and release protein components of the complement pathways, as well as secreting and anchoring ultra-large von Willebrand factor (ULVWF multimers in long string-like structures that initiate platelet adhesion during hemostasis and thrombosis. The alternative complement pathway (AP is an important non-antibody-requiring host defense system. Thrombotic microangiopathies can be associated with defective regulation of the AP (atypical hemolytic-uremic syndrome or with inadequate cleavage by ADAMTS-13 of ULVWF multimeric strings secreted by/anchored to ECs (thrombotic thrombocytopenic purpura. Our goal was to determine if EC-anchored ULVWF strings caused the assembly and activation of AP components, thereby linking two essential defense mechanisms. METHODOLOGY/PRINCIPAL FINDINGS: We quantified gene expression of these complement components in cultured human umbilical vein endothelial cells (HUVECs by real-time PCR: C3 and C5; complement factor (CF B, CFD, CFP, CFH and CFI of the AP; and C4 of the classical and lectin (but not alternative complement pathways. We used fluorescent microscopy, monospecific antibodies against complement components, fluorescent secondary antibodies, and the analysis of >150 images to quantify the attachment of HUVEC-released complement proteins to ULVWF strings secreted by, and anchored to, the HUVECs (under conditions of ADAMTS-13 inhibition. We found that HUVEC-released C4 did not attach to ULVWF strings, ruling out activation of the classical and lectin pathways by the strings. In contrast, C3, FB, FD, FP and C5, FH and FI attached to ULVWF strings in quantitative patterns consistent with assembly of the AP components into active complexes. This was verified when non-functional FB blocked the formation of AP C3 convertase complexes (C3bBb on ULVWF strings. CONCLUSIONS/SIGNIFICANCE: AP components are assembled and activated on EC-secreted/anchored ULVWF multimeric

  15. The Anchoring Effects in Differential Sources of Anchor Information:How Experimenter-Provided Anchors and Self-generated Anchors Affect the Dual-Processing Model%内在锚与外在锚对锚定效应及其双加工机制的影响

    Institute of Scientific and Technical Information of China (English)

    李斌; 徐富明; 张军伟; 刘腾飞; 蒋多; 邓子鹃

    2012-01-01

    The anchoring effect, one of the most common heuristics, refers to a phenomenon that the assimilation of a quantitative estimates a previously presented irrelative number under uncertain circumstances. Since the anchoring effect was proposed by Tversky and Kahneman in 1974 , lots of research has been done about it. Now there are two main models while account for anchoring effects:the anchoring and adjustment heuristic and the selective accessibility model. But most of researches just focused on the anchoring and adjustment heuristic or the selective accessibility model separately. In recent years, some scholars have pointed out that even if participants have a self-generated anchor, when there is an experimenter-provided anchor showing up at the same time, the processing mechanism of the anchoring effect is not only anchoring and adjustment heuristic, but also may happen to the selective accessibility model at the same time. But no research has been available to prove it yet. Therefore, this study is to try to explore the relationship between anchoring and adjustment heuristic and the selective accessibility model with the self-generated anchor and the experimenter-provided anchor showing up at the same time. A total of 64 undergraduate or graduate students were recruited and paid for their participation. The participants, were placed in separate cubicles in the laboratory, each containing a computer used to present the information and to register the dependent measures. The participants were told that the experiment was about how people made estimates. First, they had to estimate if there was predominance of capital or lower case letters in several letter combinations. After that, they had to make some probability estimates. The letter combination task was used to conceal the true purpose of our experiment. In this task, the participants had to estimate if a letter combination of six letters (e.g. MJFqRe) contained more capital letters or more lower case

  16. RhoA Kinase (Rock) and p90 Ribosomal S6 Kinase (p90Rsk) phosphorylation of the sodium hydrogen exchanger (NHE1) is required for lysophosphatidic acid-induced transport, cytoskeletal organization and migration.

    Science.gov (United States)

    Wallert, Mark A; Hammes, Daniel; Nguyen, Tony; Kiefer, Lea; Berthelsen, Nick; Kern, Andrew; Anderson-Tiege, Kristina; Shabb, John B; Muhonen, Wallace W; Grove, Bryon D; Provost, Joseph J

    2015-03-01

    The sodium hydrogen exchanger isoform one (NHE1) plays a critical role coordinating asymmetric events at the leading edge of migrating cells and is regulated by a number of phosphorylation events influencing both the ion transport and cytoskeletal anchoring required for directed migration. Lysophosphatidic acid (LPA) activation of RhoA kinase (Rock) and the Ras-ERK growth factor pathway induces cytoskeletal reorganization, activates NHE1 and induces an increase in cell motility. We report that both Rock I and II stoichiometrically phosphorylate NHE1 at threonine 653 in vitro using mass spectrometry and reconstituted kinase assays. In fibroblasts expressing NHE1 alanine mutants for either Rock (T653A) or ribosomal S6 kinase (Rsk; S703A) we show that each site is partially responsible for the LPA-induced increase in transport activity while NHE1 phosphorylation by either Rock or Rsk at their respective site is sufficient for LPA stimulated stress fiber formation and migration. Furthermore, mutation of either T653 or S703 leads to a higher basal pH level and a significantly higher proliferation rate. Our results identify the direct phosphorylation of NHE1 by Rock and suggest that both RhoA and Ras pathways mediate NHE1-dependent ion transport and migration in fibroblasts.

  17. NDE applications in an anchor bolt and sign bridge inspection program

    Science.gov (United States)

    Fish, Philip E.

    1998-03-01

    Wisconsin Department of Transportation implemented an Anchor Bolt Inspection Program and Sign Bridge Inspection Program in 1990 for cantilever & overhead sign structures, high mast light towers, interstate light towers, and signal masts. All cantilever & overhead sign structures and high mast light towers have been inspected in-depth 'hands on' during this time period while implementation of inspection continues for interstate light towers and signal masts. The program requires an experienced inspection team and a practical inspection approach. Inspection preparation includes review of all background information such as design plans, design computations, construction plans, shop plans, and maintenance history. An inspection plan is developed. Special emphasis is placed on determining type of structure, type of material, welding details. For anchor bolts, type of material, cut or rolled threads, type of coating, and method of placement are important issues. Inspection emphasis are on 'hands on' and Nondestructive evaluation. Special emphasis is placed visual conditions of sign structures and anchor bolts (cut or rolled threads, straightness, corrosion, nut tension, etc.) along with ultrasonic inspection. This program places a strong emphasis on Non Destructive Testing (NDT), especially ultrasonic. Procedures and inspection calibrations for anchor bolts, are developed from similar anchor bolt geometry and material type. Cut notches are placed in the anchor bolts at locations of possible failure. NDT inspection calibrations are performed from these bolts. Report documentation includes all design plans, pictorial documentation of structural deficiencies, sketches, nondestructive evaluation reports, conclusions, and recommendations. This program has been successful in locating critical cracks and failed members on existing sign structures and new sign structures less than six months old. Also, failed anchor bolts and critical cracks have been located on high mast light

  18. Murasaki: a fast, parallelizable algorithm to find anchors from multiple genomes.

    Directory of Open Access Journals (Sweden)

    Kris Popendorf

    Full Text Available BACKGROUND: With the number of available genome sequences increasing rapidly, the magnitude of sequence data required for multiple-genome analyses is a challenging problem. When large-scale rearrangements break the collinearity of gene orders among genomes, genome comparison algorithms must first identify sets of short well-conserved sequences present in each genome, termed anchors. Previously, anchor identification among multiple genomes has been achieved using pairwise alignment tools like BLASTZ through progressive alignment tools like TBA, but the computational requirements for sequence comparisons of multiple genomes quickly becomes a limiting factor as the number and scale of genomes grows. METHODOLOGY/PRINCIPAL FINDINGS: Our algorithm, named Murasaki, makes it possible to identify anchors within multiple large sequences on the scale of several hundred megabases in few minutes using a single CPU. Two advanced features of Murasaki are (1 adaptive hash function generation, which enables efficient use of arbitrary mismatch patterns (spaced seeds and therefore the comparison of multiple mammalian genomes in a practical amount of computation time, and (2 parallelizable execution that decreases the required wall-clock and CPU times. Murasaki can perform a sensitive anchoring of eight mammalian genomes (human, chimp, rhesus, orangutan, mouse, rat, dog, and cow in 21 hours CPU time (42 minutes wall time. This is the first single-pass in-core anchoring of multiple mammalian genomes. We evaluated Murasaki by comparing it with the genome alignment programs BLASTZ and TBA. We show that Murasaki can anchor multiple genomes in near linear time, compared to the quadratic time requirements of BLASTZ and TBA, while improving overall accuracy. CONCLUSIONS/SIGNIFICANCE: Murasaki provides an open source platform to take advantage of long patterns, cluster computing, and novel hash algorithms to produce accurate anchors across multiple genomes with

  19. LRD-22, a novel dual dithiocarbamatic acid ester, inhibits Aurora-A kinase and induces apoptosis and cell cycle arrest in HepG2 cells

    Energy Technology Data Exchange (ETDEWEB)

    Wang, Huiling; Li, Ridong [State Key Laboratory of Natural and Biomimetic Drugs, School of Pharmaceutical Sciences, Peking University, Beijing (China); Li, Li [Department of Cell Biology, School of Basic Medical Sciences, Peking University, Beijing (China); Ge, Zemei [State Key Laboratory of Natural and Biomimetic Drugs, School of Pharmaceutical Sciences, Peking University, Beijing (China); Zhou, Rouli, E-mail: rlzhou@bjmu.edu.cn [Department of Cell Biology, School of Basic Medical Sciences, Peking University, Beijing (China); Li, Runtao, E-mail: lirt@bjmu.edu.cn [State Key Laboratory of Natural and Biomimetic Drugs, School of Pharmaceutical Sciences, Peking University, Beijing (China)

    2015-02-27

    In this study we investigated the antitumor activity of the novel dual dithiocarbamatic acid ester LRD-22 in vitro and in vivo. Several cancer cell lines were employed to determine the effect of LRD-22 on cell growth, and the MTT assay showed there was a significant decrease in viable tumor cell numbers in the presence of LRD-22, especially in the HepG2 cell line. Colony formation assay also showed LRD-22 strongly inhibits HepG2 cell growth. Evaluation of the mechanism involved showed that inhibitory effects of LRD-22 on cell growth are due to induction of apoptosis and G2/M arrest. LRD-22 inhibited Aurora-A phosphorylation at Thr{sub 288} and subsequently impaired p53 phosphorylation at Ser{sub 315} which was associated with the proteasome degradation pathway. Tumor suppressor protein p53 is stabilized by this mechanism and accumulates through inhibition of Aurora-A kinase activity via treatment with LRD-22. In vivo study of HepG2 xenograft in nude mice also shows LRD-22 suppresses tumor growth at a concentration of 5 mg/kg without animals suffering loss of body weight. In conclusion, our results demonstrate LRD-22 acts as an Aurora-A kinase inhibitor to induce apoptosis and inhibit proliferation in HepG2 cells, and should be considered as a promising targeting agent for HCC therapy. - Highlights: • LRD-22 significantly inhibits cancer cell growth, especially in the HepG2 cell line. • The inhibitory effect of LRD-22 is due to induction of apoptosis and cell cycle arrest. • LRD-22 inhibits Aurora-A phosphorylation which results in subsequent impairment of the p53 pathway. • LRD-22 suppresses tumor growth in xenograft mice without body weight loss.

  20. DEVELOPMENT OF DUAL ANCHORED SHEET PILE WALL METHOD TO INCREASE FRONT WATER DEPTH AND SEISMIC RESISTANCE OF EXISTING QUAY

    Science.gov (United States)

    Nakamura, Yasushi; Sato, Masakatsu; Kikuchi, Yoshiaki; Sugano, Takahiro; Morikawa, Yoshiyuki; Hoshino, Masami; Miki, Kenichi

    Recently the dual anchored sheet pile wall method has been developed to increase a front water depth and seismic resistance of existing quay walls by providing an additional anchor in the lower level of them to reduce a flexural moment of the sheet piles and a tension of the anchors. The existing technical information is not enough to evaluate the seismic behavior and the retrofit of the quay walls with anchors at two different levels. Therefore the experiments with a scale model set on the vibration table of the centrifugal apparatus as well as two dimensional effective stress analyses have been mobilized to investigate the seismic retrofit of the dual anchored sheet pile wall. The experiments and analyses demonstrate the increase the earthquake resistance of quay walls, because they showed the additional anchor can reduce the stress of the sheet walls to one half.

  1. Anchoring ethinylestradiol induced gene expression changes with testicular morphology and reproductive function in the medaka.

    Directory of Open Access Journals (Sweden)

    Hilary D Miller

    Full Text Available Environmental estrogens are ubiquitous in the environment and can cause detrimental effects on male reproduction. In fish, a multitude of effects from environmental estrogens have been observed including altered courting behavior and fertility, sex reversal, and gonadal histopathology. However, few studies in fish assess the impacts of estrogenic exposure on a physiological endpoint, such as reproduction, as well as the associated morphologic response and underlying global gene expression changes. This study assessed the implications of a 14 day sub-chronic exposure of ethinylestradiol (EE2; 1.0 or 10.0 µg/L EE2 on male medaka fertility, testicular histology and testicular gene expression. The findings demonstrate that a 14 day exposure to EE2 induced impaired male reproductive capacity and time- and dose-dependent alterations in testicular morphology and gene expression. The average fertilization rate/day following the exposure for control, 1.0 and 10.0 µg/L EE2 was 91.3% (±4.4, 62.8% (±8.3 and 28.8% (±5.8, respectively. The testicular morphologic alterations included increased germ cell apoptosis, decreased germinal epithelium and thickening of the interstitium. These changes were highly associated with testicular gene expression changes using a medaka-specific microarray. A pathway analysis of the differentially expressed genes emphasized genes and pathways associated with apoptosis, cell cycle and proliferation, collagen production/extracellular matrix organization, hormone signaling, male reproduction and protein ubiquitination among others. These findings highlight the importance of anchoring global gonadal gene expression changes with morphology and ultimately with tissue/organ function.

  2. Differences regarding job satisfaction and job involvement of psychologists with different dominant career anchors

    Directory of Open Access Journals (Sweden)

    CL Bester

    2006-09-01

    Full Text Available In order to contribute to higher levels of job satisfaction, job involvement , and productivity, a match or fit should be established between the dominant career anchor associated with a specific occupation and that of the employee. A career anchor is an individual’s set of self-perceived talents, abilities, motives, needs and values that form the nucleus of one’s occupational self-concept. Psychologists have always been part of the service orientated careers and therefore one would expect that it is likely that their dominant career anchor would be service orientation. If this is the case, psychologists with service as their dominant career anchor are supposed to have greater job satisfaction and job involvement compared to those with different career anchors. However, according to literature, this assumption is not necessarily correct. The primary goals of the current study were to determine whether in fact service is the dominant career anchor of psychologists in the Free State and whether there are significant differences regarding job satisfaction and job involvement between psychologists with and without service as their dominant career anchor. A third goal was to determine whether psychologists with different dominant career anchors differ significantly from one another regarding job satisfaction and job involvement. Questionnaires measuring career orientations, job satisfaction and job involvement were sent to 165 of the 171 registered psychologists in the Free State region. Only 75 psychologists (45,5% responded which exceeded the traditional return rate of 20 to 30%. Due to the small sample of respondents, a nonparametric statistical test, namely the Mann Whitney U test was conducted to determine possible differences. An analysis of the data showed that 21 respondents had entrepreneurship as their dominant career orientation while 12 fell in the technical/functional, 12 in the challenging, 9 in the service and 8 in the autonomy

  3. Comparison of the Tendon Damage Caused by Four Different Anchor Systems Used in Transtendon Rotator Cuff Repair

    OpenAIRE

    Qing-Song Zhang; Sen Liu; Qiuyang Zhang; Yun Xue; Dongxia Ge; Michael J O'Brien; Savoie, Felix H.; Zongbing You

    2012-01-01

    Objectives. The objective of this study was to compare the damage to the rotator cuff tendons caused by four different anchor systems. Methods. 20 cadaveric human shoulder joints were used for transtendon insertion of four anchor systems. The Healix Peek, Fastin RC, Bio-Corkscrew Suture, and Healix Transtend anchors were inserted through the tendons using standard transtendon procedures. The areas of tendon damage were measured. Results. The areas of tendon damage (mean ± standard deviation, ...

  4. Diagnostic and therapeutic direct peroral cholangioscopy using an intraductal anchoring balloon

    Institute of Scientific and Technical Information of China (English)

    Mansour A Parsi; Tyler Stevens; John J Vargo

    2012-01-01

    AIM:To report our experience using a recently introduced anchoring balloon for diagnostic and therapeutic direct peroral cholangioscopy (DPOC).METHODS:Consecutive patients referred for diagnostic or therapeutic peroral cholangioscopy were evaluated in a prospective cohort study.The patients underwent DPOC using an intraductal anchoring balloon,which was recently introduced to allow consistent access to the biliary tree with an ultraslim upper endoscope.The device was later voluntarily withdrawn from the market by the manufacturer.RESULTS:Fourteen patients underwent DPOC using the anchoring balloon.Biliary access with an ultraslim upper endoscope was accomplished in all 14 patients.In 12 (86%) patients,ductal access required sphincteroplasty with a 10-mm dilating balloon.Intraductal placement of the ultraslim upper endoscope allowed satisfactory visualization of the biliary mucosa to the level of the confluence of the right and left hepatic ducts in 13 of 14 patients (93%).Therapeutic interventions by DPOC were successfully completed in all five attempted cases (intraductal biopsy in one and DPOC guided laser lithotripsy in four).Adverse events occurred in a patient on immunosuppressive therapy who developed an intrahepatic biloma at the site of the anchoring balloon.This required hospitalization and antibiotics.Repeat endoscopic retrograde cholangiopancreatography 8 wk after the index procedure showed resolution of the biloma.CONCLUSION:Use of this anchoring balloon allowed consistent access to the biliary tree for performance of diagnostic and therapeutic DPOC distal to the biliary bifurcation.

  5. Interference Screw vs. Suture Anchor Fixation for Open Subpectoral Biceps Tenodesis: Does it Matter?

    Directory of Open Access Journals (Sweden)

    Gobezie Reuben

    2008-09-01

    Full Text Available Abstract Background Bioabsorbable interference screw fixation has superior biomechanical properties compared to suture anchor fixation for biceps tenodesis. However, it is unknown whether fixation technique influences clinical results. Hypothesis We hypothesize that subpectoral interference screw fixation offers relevant clinical advantages over suture anchor fixation for biceps tenodesis. Study Design Case Series. Methods We performed a retrospective review of a consecutive series of 88 patients receiving open subpectoral biceps tenodesis with either interference screw fixation (34 patients or suture anchor fixation (54 patients. Average follow up was 13 months. Outcomes included Visual Analogue Pain Scale (0–10, ASES score, modified Constant score, pain at the tenodesis site, failure of fixation, cosmesis, deformity (popeye and complications. Results There were no failures of fixation in this study. All patients showed significant improvement between their preoperative and postoperative status with regard to pain, ASES score, and abbreviated modified Constant scores. When comparing IF screw versus anchor outcomes, there was no statistical significance difference for VAS (p = 0.4, ASES score (p = 0.2, and modified Constant score (P = 0.09. One patient (3% treated with IF screw complained of persistent bicipital groove tenderness, versus four patients (7% in the SA group (nonsignificant. Conclusion Subpectoral biceps tenodesis reliably relieves pain and improves function. There was no statistically significant difference in the outcomes studied between the two fixation techniques. Residual pain at the site of tenodesis may be an issue when suture anchors are used in the subpectoral location.

  6. A Study on the Holding Capacity Safety Factors for Torpedo Anchors

    Directory of Open Access Journals (Sweden)

    Luís V. S. Sagrilo

    2012-01-01

    Full Text Available The use of powerful numerical tools based on the finite-element method has been improving the prediction of the holding capacity of fixed anchors employed by the offshore oil industry. One of the main achievements of these tools is the reduction of the uncertainty related to the holding capacity calculation of these anchors. Therefore, it is also possible to reduce the values of the associated design safety factors, which have been calibrated relying on models with higher uncertainty, without impairing the original level of structural safety. This paper presents a study on the calibration of reliability-based safety factors for the design of torpedo anchors considering the statistical model uncertainty evaluated using results from experimental tests and their correspondent finite-element-based numerical predictions. Both working stress design (WSD and load and resistance factors design (LRFD design methodologies are investigated. Considering the WSD design methodology, the single safety is considerably lower than the value typically employed in the design of torpedo anchors. Moreover, a LRFD design code format for torpedo anchors is more appropriate since it leads to designs having less-scattered safety levels around the target value.

  7. Synergism Analysis of Bedding Slope with Piles and Anchor Cable Support under Sine Wave Vehicle Load

    Directory of Open Access Journals (Sweden)

    Li Dan-Feng

    2016-01-01

    Full Text Available Slope instability under dynamic load is the technical difficulty in the engineering; the evaluation of slope stability under dynamic load and the control of dynamic load is particularly important. In this paper, taking the right side slope of K27+140 m~380 m typical section (K27 slope for short in Chongqing Fuling-Fengdu-Shizhu expresses highway as an example to calculate and analyze. The K27 slope is under sinusoidal vehicle load and supported by anchor cable and antislide pile to resist downslide strength; at the same time, the combined effect of them is studied. Three-dimensional finite element methodology (FEM is used to simulate the bedding slope with piles and anchor cable support; furthermore, the eigenvalue can be obtained. In order to reduce error of the elastic boundary conditions caused by the reflection effect of wavelengths, the combination of Lysmer surface viscous boundary and traditional ground support boundaries is utilized to analyze and calculate the time-histories during bedding slope under dynamic load. The dynamic response of pile anchor support to resist sliding force is obtained. The concept of the pile anchor supporting coordinate interval is put forward. Furthermore, it is verified that the pile anchor supporting coordinate interval can be used to evaluate the stability of the slope under dynamic load and provide a new method for the control of the dynamic load.

  8. COMPLICATIONS RESULTING FROM THE USE OF METAL ANCHORS IN SHOULDER ARTHROSCOPY

    Science.gov (United States)

    Godinho, Glaydson Gomes; França, Flavio Oliveira; Alves Freitas, José Marcio; Aguiar, Paulo Nascimento; de Carvalho Leite, Marcelo

    2015-01-01

    To identify the complications concerning the use of metal anchors in shoulder arthroscopic procedures. Methods: 28 shoulders of 28 patients (23 male and 5 female) have been re-operated in the period between December 1997 and August 2007, at Hospital Ortopédico, Belo Horizonte Hospital and Military Police Hospital in Belo Horizonte, MG, as a result of complications such as loose anchors and prominent anchors. The primary surgeries intended to treat 20 anterior traumatic instabilities (71.5%), one posterior instability (3.5%), one slap injury (3.5%), six procedures for treating injuries on the rotator cuff (21.5%). We used the X-ray classification suggested by Samilson and Prieto and Outerbridge arthroscopic classification for assessing patients' degree of arthrosis. All patients were evaluated by the UCLA (University of California at Los Angeles) index criteria. Results: In all patients, arthroscopic reviews were made. In two cases, after anchors removal, clinical signs of instability were seen, leading to the decision of providing open stabilization by Latarjet-Patte technique. Conclusion: the complications with metallic-suture anchors result from inappropriate surgical techniques applied in arthroscopy. PMID:26998465

  9. Mechanical mechanism analysis of tension type anchor based on shear displacement method

    Institute of Scientific and Technical Information of China (English)

    XIAO Shu-jun; CHEN Chang-fu

    2008-01-01

    Based on the fact that the shear stress along anchorage segment is neither linearly nor uniformly distributed, the load transfer mechanism of the tension type anchor was studied and the mechanical characteristic of anchorage segment was analyzed. Shear stress-strain relationship of soil surrounding anchorage body was simplified into three-folding-lines model consisting of elastic phase, elasto-plastic phase and residual phase considering its softening eharaeteristic. Meanwhile, shear displacement method that has been extensively used in the analysis of pile foundation was introduced. Based on elasto-plastic theory, the distributions of displacement, shear stress and axial force along the anchorage segment of tension type anchor were obtained, and the formula for calculating the elastic limit load was also developed accordingly. Finally, an example was given to discuss the variation of stress and displacement in the anchorage segment with the loads exerted on the anchor, and a program was worked out to calculate the anchor maximum bearing capacity. The influence of some parameters on the anchor bearing capacity was discussed, and effective anchorage length was obtained simultaneously. The results show that the shear stress first increases and then decreases and finally trends to theresidual strength with increase of distance from bottom of the anchorage body, the displacement increases all the time with theincrease of distance from bottom of the anchorage body, and the increase of velocity gradually becomes greater.

  10. Tunable Signal Processing through a Kinase Control Cycle: the IKK Signaling Node

    Science.gov (United States)

    Behar, Marcelo; Hoffmann, Alexander

    2013-01-01

    The transcription factor NFκB, a key component of the immune system, shows intricate stimulus-specific temporal dynamics. Those dynamics are thought to play a role in controlling the physiological response to cytokines and pathogens. Biochemical evidence suggests that the NFκB inducing kinase, IKK, a signaling hub onto which many signaling pathways converge, is regulated via a regulatory cycle comprising a poised, an active, and an inactive state. We hypothesize that it operates as a modulator of signal dynamics, actively reshaping the signals generated at the receptor proximal level. Here we show that a regulatory cycle can function in at least three dynamical regimes, tunable by regulating a single kinetic parameter. In particular, the simplest three-state regulatory cycle can generate signals with two well-defined phases, each with distinct coding capabilities in terms of the information they can carry about the stimulus. We also demonstrate that such a kinase cycle can function as a signal categorizer classifying diverse incoming signals into outputs with a limited set of temporal activity profiles. Finally, we discuss the extension of the results to other regulatory motifs that could be understood in terms of the regimes of the three-state cycle. PMID:23823243

  11. Aurora A Kinase Contributes to a Pole-Based Error Correction Pathway.

    Science.gov (United States)

    Ye, Anna A; Deretic, Jovana; Hoel, Christopher M; Hinman, Albert W; Cimini, Daniela; Welburn, Julie P; Maresca, Thomas J

    2015-07-20

    Chromosome biorientation, where sister kinetochores attach to microtubules (MTs) from opposing spindle poles, is the configuration that best ensures equal partitioning of the genome during cell division. Erroneous kinetochore-MT attachments are commonplace but are often corrected prior to anaphase. Error correction, thought to be mediated primarily by the centromere-enriched Aurora B kinase (ABK), typically occurs near spindle poles; however, the relevance of this locale is unclear. Furthermore, polar ejection forces (PEFs), highest near poles, can stabilize improper attachments by pushing mal-oriented chromosome arms away from spindle poles. Hence, there is a conundrum: erroneous kinetochore-MT attachments are weakened where PEFs are most likely to strengthen them. Here, we report that Aurora A kinase (AAK) opposes the stabilizing effect of PEFs. AAK activity contributes to phosphorylation of kinetochore substrates near poles and its inhibition results in chromosome misalignment and an increased incidence of erroneous kinetochore-MT attachments. Furthermore, AAK directly phosphorylates a site in the N-terminal tail of Ndc80/Hec1 that has been implicated in reducing the affinity of the Ndc80 complex for MTs when phosphorylated. We propose that an AAK activity gradient contributes to correcting mal-oriented kinetochore-MT attachments in the vicinity of spindle poles.

  12. RhVI1 is a membrane-anchored vacuolar invertase highly expressed in Rosa hybrida L. petals.

    Science.gov (United States)

    Farci, Domenica; Collu, Gabriella; Kirkpatrick, Joanna; Esposito, Francesca; Piano, Dario

    2016-05-01

    Invertases are a widespread group of enzymes that catalyse the conversion of sucrose into fructose and glucose. Plants invertases and their substrates are essential factors that play an active role in primary metabolism and in cellular differentiation and by these activities they sustain development and growth. Being naturally present in multiple isoforms, invertases are known to be highly differentiated and tissue specific in such a way that every isoform is characteristic of a specific part of the plant. In this work, we report the identification of the invertase RhVI1 that was found to be highly expressed in rose petals. A characterization of this protein revealed that RhVI1 is a glycosylated membrane-anchored protein associated with the cytosolic side of the vacuolar membrane which occurs in vivo in a monomeric form. Purification yields have shown that the levels of expression decreased during the passage of petals from buds to mature and pre-senescent flowers. Moreover, the activity assay indicates RhVI1 to be an acidic vacuolar invertase. The physiological implications of these findings are discussed, suggesting a possible role of this protein during anthesis.

  13. Equivalent Culture-Anchored Units Translation? The Phraseological Units Issue

    Directory of Open Access Journals (Sweden)

    Maciej Paweł Jaskot

    2016-12-01

    Full Text Available Equivalent Culture-Anchored Units Translation? The Phraseological Units Issue This article examines a question that has been of long-standing interest to linguists working in the fields of cross-linguistic phraseology and the translation of idiosyncratic language units, such as phraseological units (PUs. The challenge of translating PUs, which are understood as "patterns sanctioned by a given culture", involves the translation of culturemes. Therefore, a good translator must be able to assess the importance of the elements containing cultural references in the source language while "moving" them to the target language. When translating PUs, it is desirable that interlingual (cross-linguistic equivalence be achieved. The fact that translations of a PU can be very different (the translator can paraphrase the text, creatively change it, or simply eliminate the PU implies that the translational equivalence of PUs must be functional. While a cross-linguistic comparison (and the achievement of translational equivalence of PUs can be made by omitting the form parameter, it is desirable to preserve the extension and semantic structure, the connotative-pragmatic component, and the phrase combinatorics.   Ekwiwalencja jedostek "kulturowo zakotwiczonych"? Kwestia jednostek frazeologicznych Artykuł porusza zagadnienie, które od kilkudziesięciu lat cieszy się zainteresowaniem językoznawców zajmujących się konfrontacją językową frazeologii oraz tłumaczeniem idiosynkratycznych jednostek językowych, takich jak jednostki frazeologiczne (JF. Wyzwaniem podczas tłumaczenia JF, rozumianych jako "wzorce sankcjonowane przez daną kulturę", jawi się konieczność tłumaczenia kulturemów. Tłumacz zatem stoi w obliczu konieczności prawidłowej oceny danego elementu pod względem jego odniesień kulturowych w języku źródłowym podczas przekładania JF na język docelowy. Podczas tłumaczenia JF pożądane jest osiągnięcie międzyjęzykowej (cross

  14. Protein raftophilicity. How bioinformatics can help membranologists

    DEFF Research Database (Denmark)

    Nielsen, Henrik; Sperotto, Maria Maddalena

    for bioprocesses. Protein raftophilicity depends on features such as lipidation and GPI-anchoring. Can this affinity be inferred solely by knowing such features, without knowing the physical and physico-chemical properties of biomembranes? We tried to answer the question by an artificial neural network (ANN...

  15. ANALYSIS ON INCREMENTAL COHESION OF SURROUNDING ROCK DUE TO PRESTRESSED CABLE ANCHOR

    Institute of Scientific and Technical Information of China (English)

    夏建中; 闫莫明; 滕年保; 李方政

    1998-01-01

    A new method for determining the incremental cohesion △Cm of surrounding rock due toprestressed cable anchor is presented, and the formulas for △Cm are deduced and ACredistributions also are discussed, based on the two anchorage effects, one is the effect with theprestressed value △Q3 of cable anchor improving the stress state of surrounding rock andincreasing the surrounding rock strength, the other is the fully encapsulated effect. Thedetermined incremental cohesion △Cm is subjected to the model test and field measurement in thereferences, and coincides well with those tested results. The formulas for △Cm can be used indesigning supoport parameters and related numerical analyses of prestressed cable anchor.

  16. Traction endurance biomechanical study of metallic suture anchors at different insertion angles

    Directory of Open Access Journals (Sweden)

    Azato Flávia Namie

    2003-01-01

    Full Text Available The suture anchors' insertion angle and its traction resistance are the main subjects of this study. Twenty trials were realized using threaded suture anchors in four diferents angulations (30º /45º /60º /90º in human bone (distal femur and another twenty trials in artificial bone (SawboneTM. The anchors were pulled out being tractioned uprightly from its bone surface by a Kratos Universal test machine. The human bone results found no relation between the main subjects of this study, so whithout statistical value. On the other hand at the artificial bone the insertion angle of 90º beared more traction, being statistically significant compared to the other angles.

  17. Computing of the Anchor by the Method of Three-Dimension Point-Radiate Infinite Elements

    Institute of Scientific and Technical Information of China (English)

    2007-01-01

    On the basis of the one-dimension infinite element theory, the coordinate translation and shape function of 3D point-radiate 8-node and 4-node infinite elements are derived. They are coupled with 20-node and 8-node finite elements to compute the compression distortion of the prestressed anchorage segment. The results indicate that when the prestressed force acts on the anchorage head and segment, the stresses and the displacements in the rock around the anchorage head and segment concentrate on the zone center with the anchor axis, and they decrease with exponential forms. Therefore,the stresses and the displacement spindles are formed. The calculating results of the infinite element are close to the theoretical results. This indicates the method is right. This article introduces a new way to study the mechanism of prestressed anchors. The obtained results have an important role in the research of the anchor mechanism and engineering application.

  18. Fluid-structure interaction in straight pipelines with different anchoring conditions

    Science.gov (United States)

    Ferras, David; Manso, Pedro A.; Schleiss, Anton J.; Covas, Dídia I. C.

    2017-04-01

    This investigation aims at assessing the fluid-structure interaction (FSI) occurring during hydraulic transients in straight pipeline systems fixed to anchor blocks. A two mode 4-equation model is implemented incorporating the main interacting mechanisms: Poisson, friction and junction coupling. The resistance to movement due to inertia and dry friction of the anchor blocks is treated as junction coupling. Unsteady skin friction is taken into account in friction coupling. Experimental waterhammer tests collected from a straight copper pipe-rig are used for model validation in terms of wave shape, timing and damping. Numerical results successfully reproduce laboratory measurements for realistic values of calibration parameters. The novelty of this paper is the presentation of a 1D FSI solver capable of describing the resistance to movement of anchor blocks and its effect on the transient pressure wave propagation in straight pipelines.

  19. The anchoring function: parental authority and the parent-child bond.

    Science.gov (United States)

    Omer, Haim; Steinmetz, Sarit G; Carthy, Tal; von Schlippe, Arist

    2013-06-01

    Descriptions of parental authority and of the formation of a secure parent-child bond have remained unconnected in conceptualizations about parenting and child development. The parental anchoring function is here presented as an integrative metaphor for the two fields. Parents who fulfill an anchoring function offer a secure relational frame for the child, while also manifesting a stabilizing and legitimate kind of authority. The anchoring function enriches the two fields by: (1) adding a dimension of authority to the acknowledged functions of the safe haven and the secure base that are seen as core to a secure parent-child bond, and (2) adding considerations about the parent-child bond to Baumrind's classical description of authoritative parenting.

  20. Anchoring Strength of Thin Aligned-Polymer Films Formed by Liquid Crystalline Monomer

    Science.gov (United States)

    Murashige, Takeshi; Fujikake, Hideo; Ikehata, Seiichiro; Sato, Fumio

    2003-04-01

    We have evaluated the polar anchoring strength of a thin molecule-aligned polymer film formed by a liquid crystalline monomer. The polymer film was obtained by photopolymerization of the monomer oriented by a rubbed polyimide alignment layer in a chamber filled with N2 gas. We fabricated a nematic liquid crystal cell using the thin aligned-polymer films as alignment layers, and then evaluated the anchoring strength of the polymer by measuring the optical retardation curve of the cell driven by voltages. The experimental result showed that the anchoring strength was one order of magnitude lower than that of a conventional rubbed polyimide alignment layer, and decreased with increasing the cure temperature of the monomer film.

  1. Analysis of Cracking Mode of Anchor Structure of Underground Engineering Induced by Reinforcement Corrosion

    Directory of Open Access Journals (Sweden)

    Wantao Ding

    2014-02-01

    Full Text Available Based on elastic theory and assumption of maximum tensile-stress failure criterion, together with construction process of anchor structure and rust expansion critical process, this study proposed a simplified reinforcement rust expansion mechanical model of anchor structure system. Elastic criterion of different initial cracking mode was rewarded under different stress ratios. According to analysis of critical cracking mode of different medium, cracking order of mortar and surrounding rock depended on their material parameters, in-situ stress and thickness of mortar cover. Critical cracking conditions of different medium without effect of in-situ stress was the same as that of considering in-situ stress while k is equal to 3 or 1/3. And engineering example shows that three different cracking modes exist under different stress ratios. The result provides a useful reference for analysis of mechanical deterioration mechanism of anchor structure and design of support structure of underground engineering.

  2. Pre-stressed anchoring beam technique applicable in the reinforcement of high-steep slopes

    Institute of Scientific and Technical Information of China (English)

    Zhifa YANG; Luqing ZHANG; Jiewang ZHU

    2006-01-01

    During the construction of some large-scale rock engineering, high-steep slopes and insufficient slope stability induced by unloading fissures are often encountered. For the reinforcement of these slopes, some techniques (including conventional pre-stressed anchoring cable and unconventional anchoring hole) are usually utilized, however, having several obvious defects. Thus, it is very difficult for a designer to design an efficient reinforcement scheme for the high-steep slopes. For this reason, the authors develop the pre-stressed anchoring beam technique, in which tensile capacity of pre-stressed structures are fully utilized. It is analyzed that the new technique is characterized by multi-functions, including engineering investigation, efficient reinforcement, drainage, monitoring and urgent strength supplement, and hoped to be extensively applicable in the reinforcement of high-steep slopes.

  3. Mussel-Inspired Anchoring of Polymer Loops That Provide Superior Surface Lubrication and Antifouling Properties.

    Science.gov (United States)

    Kang, Taegon; Banquy, Xavier; Heo, Jinhwa; Lim, Chanoong; Lynd, Nathaniel A; Lundberg, Pontus; Oh, Dongyeop X; Lee, Han-Koo; Hong, Yong-Ki; Hwang, Dong Soo; Waite, John Herbert; Israelachvili, Jacob N; Hawker, Craig J

    2016-01-26

    We describe robustly anchored triblock copolymers that adopt loop conformations on surfaces and endow them with unprecedented lubricating and antifouling properties. The triblocks have two end blocks with catechol-anchoring groups and a looping poly(ethylene oxide) (PEO) midblock. The loops mediate strong steric repulsion between two mica surfaces. When sheared at constant speeds of ∼2.5 μm/s, the surfaces exhibit an extremely low friction coefficient of ∼0.002-0.004 without any signs of damage up to pressures of ∼2-3 MPa that are close to most biological bearing systems. Moreover, the polymer loops enhance inhibition of cell adhesion and proliferation compared to polymers in the random coil or brush conformations. These results demonstrate that strongly anchored polymer loops are effective for high lubrication and low cell adhesion and represent a promising candidate for the development of specialized high-performance biomedical coatings.

  4. A general pipeline for the development of anchor markers for comparative genomics in plants

    Directory of Open Access Journals (Sweden)

    Stougaard Jens

    2006-08-01

    Full Text Available Abstract Background Complete or near-complete genomic sequence information is presently only available for a few plant species representing a large phylogenetic diversity among plants. In order to effectively transfer this information to species lacking sequence information, comparative genomic tools need to be developed. Molecular markers permitting cross-species mapping along co-linear genomic regions are central to comparative genomics. These "anchor" markers, defining unique loci in genetic linkage maps of multiple species, are gene-based and possess a number of features that make them relatively sparse. To identify potential anchor marker sequences more efficiently, we have established an automated bioinformatic pipeline that combines multi-species Expressed Sequence Tags (EST and genome sequence data. Results Taking advantage of sequence data from related species, the pipeline identifies evolutionarily conserved sequences that are likely to define unique orthologous loci in most species of the same phylogenetic clade. The key features are the identification of evolutionarily conserved sequences followed by automated design of intron-flanking Polymerase Chain Reaction (PCR primer pairs. Polymorphisms can subsequently be identified by size- or sequence variation of PCR products, amplified from mapping parents or populations. We illustrate our procedure in legumes and grasses and exemplify its application in legumes, where model plant studies and the genome- and EST-sequence data available have a potential impact on the breeding of crop species and on our understanding of the evolution of this large and diverse family. Conclusion We provide a database of 459 candidate anchor loci which have the potential to serve as map anchors in more than 18,000 legume species, a number of which are of agricultural importance. For grasses, the database contains 1335 candidate anchor loci. Based on this database, we have evaluated 76 candidate anchor loci

  5. Limit span of self-anchored cable-stayed suspension cooperation system bridge based on strength

    Institute of Scientific and Technical Information of China (English)

    Zhe ZHANG; Huili WANG; Sifeng QIN; Xiaomeng GE

    2009-01-01

    The limit span of self-anchored cable-stayed suspension bridge is deduced. The relations among the geometrical parameters, loads and material characteristics are also analyzed. Based on the material strength and commonly used materials, the limit spans of self-anchored cable-stayed suspension bridges with concrete girder or steel girder under vertical static load are discussed in detail.The corresponding upper limit spans and the effect of the factors on the span are given. The results indicate that increasing rise-span ratio, height-span ratio and cable-stayed segment length or reducing the second dead load could increase the cooperation system span.

  6. Determining the penetration resistance of a cometary surface by using data from the Philae anchoring harpoon

    Science.gov (United States)

    Kargl, Günter; Kömle, Norbert I.; Macher, Wolfgang

    2014-05-01

    On November 11 2014 ESA's Rosetta spacecraft will deliver the Lander PHILAE to the surface of comet Churyumov-Gerasimenko. To fix the lander safely on the surface and allow for in situ operation of instruments like the SD2 drill or the MUPUS experiment the spacecraft will be anchored to the surface by a harpoon-type device. In addition to the anchoring function the projectile shot into the surface contains two sensors, which will be used to obtain information on the thermo-physical properties of the cometary ice: a shock accelerometer and a temperature sensor. The former will record the deceleration history of the anchoring projectile during the penetration phase. From these data information on mechanical strength of the near surface cometary material and its variation with depth can be retrieved by using appropriate modelling approaches. The temperature sensor will measure temperature variations at the depth were the anchor finally has come to rest over the lifetime of the lander at the surface. Both sensors are part of the MUPUS experiment on Philae, which is devoted to the measurement of thermo-mechanical properties. In this paper we will give a short overview on the main features of the harpoon-anchoring system in general and on the mathematical methods to be used for the evaluation of the measurement data recorded by the anchor sensors, with emphasis on the accelerometer data. An example from a test shot performed during the development phase of the Philae anchoring system is shown in the figure below. PIC Figure: Typical acceleration/deceleration signal recorded during a test shot of the anchor into a foam glass analogue material (upper left panel). The different phases of the shot (acceleration phase, free flight phase, deceleration phase) are clearly separated in this shot. Integration and double integration of the data (upper right and lower left panel) gives the velocity and depth profile versus time. This procedure allows finally to derive the

  7. Technical note: Endoscopic resection of a dermoid cyst anchored to the anterior optic chiasm

    Directory of Open Access Journals (Sweden)

    Yuichiro Yoneoka, MD, PhD

    2014-06-01

    Conclusion: To the best of our knowledge, ours is the only case of a dermoid cyst anchored to the anterior optic chiasma, which was visually confirmed under endoscopic observation. After surgery, the patient presented a transient impairment of the visual field, which was not evident at four month follow-up. It will contribute to a similar case, in which surgeons hesitate to make an incision in the optic chiasm. A subtotal excision should be considered in cases of dermoid cysts anchored to the anterior optic chiasm, because all the previously reported cases of suprasellar dermoid cysts are young people or those who have a relatively long life expectancy.

  8. Interaction of Rupture Zones of Adjacent Anchor Plates in an Analogical Medium

    Directory of Open Access Journals (Sweden)

    H. Abbad

    2013-12-01

    Full Text Available In this paper, an experimental study which required the design and implementation of a model containing plastic granules powder to simulate a natural environment, is presented. The latter is subjected to the removal of "anchor plates." For each test, several digital photographs are taken to materialize different deformed configurations during the pullout process. These photos processed in couples by the 7D software (image correlation giving the evolution of the displacement field and plane strain analogical environment. Particular attention is paid to the discussion of the interference of rupture zones of neighboring anchors by reducing the axis between plates.

  9. Photosensitizer anchored gold nanorods for targeted combinational photothermal and photodynamic therapy.

    Science.gov (United States)

    Tham, Huijun Phoebe; Chen, Hongzhong; Tan, Yu Hui; Qu, Qiuyu; Sreejith, Sivaramapanicker; Zhao, Lingzhi; Venkatraman, Subbu S; Zhao, Yanli

    2016-07-07

    Silylated zinc phthalocyanine (ZnPc) was anchored onto silica-coated gold nanorods (AuNR) with retained local surface plasmon resonance (LSPR). Independent LSPR and singlet oxygen production of anchored ZnPc enhance the photothermal and photodynamic efficacy of the obtained AuNR-Si-ZnPc under NIR light excitation. AuNR-Si-ZnPc was further grafted with hyaluronic acid (HA). Since HA has selective targeting capability to CD44 antigens, the final hybrid could target cancer cells directly for synergistic photothermal and photodynamic therapy.

  10. Focused suggestion with somatic anchoring technique: rapid self-hypnosis for pain management.

    Science.gov (United States)

    Donatone, Brooke

    2013-04-01

    This article details a self-hypnosis technique designed to teach patients how to manage acute or chronic pain through directed focus. The focused suggestion with somatic anchoring technique has been used with various types of pain, including somatic pain (arthritis, post-injury pain from bone breaks, or muscle tears), visceral pain (related to irritable bowel disease), and neuropathic pain (related to multiple sclerosis). This technique combines cognitive restructuring and mindfulness meditation with indirect and direct suggestions during hypnosis. The case examples demonstrate how the focused suggestion with somatic anchoring technique is used with both acute and chronic pain conditions when use of long-term medication has been relatively ineffective.

  11. The Roco protein family : a functional perspective

    NARCIS (Netherlands)

    Marin, Ignacio; van Egmond, Wouter N.; van Haastert, Peter J. M.

    2008-01-01

    In this review, we discuss the evolutionary, biochemical, and functional data available for members of the Roco protein family. They are characterized by having a conserved supradomain that contains a Ras-like GTPase domain, called Roc, and a characteristic COR (C-terminal of Roc) domain. A kinase d

  12. A novel membrane-anchored cytochrome c-550 of alkaliphilic Bacillus clarkii K24-1U: expression, molecular features and properties of redox potential.

    Science.gov (United States)

    Ogami, Shinichi; Hijikata, Shoichi; Tsukahara, Tamotsu; Mie, Yasuhiro; Matsuno, Toshihide; Morita, Naoki; Hara, Isao; Yamazaki, Koji; Inoue, Norio; Yokota, Atsushi; Hoshino, Tamotsu; Yoshimune, Kazuaki; Yumoto, Isao

    2009-05-01

    A membrane-anchored cytochrome c-550, which is highly expressed in obligately alkaliphilic Bacillus clarkii K24-1U, was purified and characterized. The protein contained a conspicuous sequence of Gly(22)-Asn(34), in comparison with the other Bacillus small cytochromes c. Analytical data indicated that the original and lipase-treated intermediate forms of cytochrome c-550 bind to fatty acids of C(15), C(16) and C(17) chain lengths and C(15) chain length, respectively, and it was considered that these fatty acids are bound to glycerol-Cys(18). Since there was a possibility that the presence of a diacylglycerol anchor contributed to the formation of dimeric states of this protein (20 and 17 kDa in SDS-PAGE), a C18M (Cys(18) --> Met)-cytochrome c-550 was constructed. The molecular mass of the C18M-cytochrome c-550 was determined as 15 and 10 kDa in SDS-PAGE and 23 kDa in blue native PAGE. The C18M-cytochrome c-550 bound with or without Triton X-100 formed a tetramer as the original cytochrome c-550 bound with Triton X-100, as determined by gel filtration. The midpoint redox potential of cytochrome c-550 as determined by redox titration was +83 mV, while that determined by cyclic voltammetric measurement was +7 mV. The above results indicate that cytochrome c-550 is a novel cytochrome c.

  13. The Role of Anchor Stations in the Validation of Earth Observation Satellite Data and Products. The Valencia and the Alacant Anchor Stations

    Science.gov (United States)

    Lopez-Baeza, Ernesto; Geraldo Ferreira, A.; Saleh-Contell, Kauzar

    Space technology facilitates humanity and science with a global revolutionary view of the Earth through the acquisition of Earth Observation satellite data. Satellites capture information over different spatial and temporal scales and assist in understanding natural climate processes and in detecting and explaining climate change. Accurate Earth Observation data is needed to describe climate processes by improving the parameterisations of different climate elements. Algorithms to produce geophysical parameters from raw satellite observations should go through selection processes or participate in inter-comparison programmes to ensure performance reliability. Geophysical parameter datasets, obtained from satellite observations, should pass a quality control before they are accepted in global databases for impact, diagnostic or sensitivity studies. Calibration and Validation, or simply "Cal/Val", is the activity that endeavours to ensure that remote sensing products are highly consistent and reproducible. This is an evolving scientific activity that is becoming increasingly important as more long-term studies on global change are undertaken, and new satellite missions are launched. Calibration is the process of quantitatively defining the system responses to known, controlled signal inputs. Validation refers to the process of assessing, by independent means, the quality of the data products derived from the system outputs. These definitions are generally accepted and most often used in the remote sensing context to refer specifically and respectively to sensor radiometric calibration and geophysical parameter validation. Anchor Stations are carefully selected locations at which instruments measure quantities that are needed to run, calibrate or validate models and algorithms. These are needed to quanti-tatively evaluate satellite data and convert it into geophysical information. The instruments collect measurements of basic quantities over a long timescale

  14. Anchoring of self-assembled plasmid DNA/ anti-DNA antibody/cationic lipid micelles on bisphosphonate-modified stent for cardiovascular gene delivery

    Directory of Open Access Journals (Sweden)

    Ma G

    2013-03-01

    Full Text Available Guilei Ma,1,# Yong Wang,1,# Ilia Fishbein,2 Mei Yu,1 Linhua Zhang,1 Ivan S Alferiev,2 Jing Yang,1 Cunxian Song,1 Robert J Levy2 1Institute of Biomedical Engineering, Chinese Academy of Medical Sciences and Peking Union Medical College, Tianjin, People's Republic of China; 2Children's Hospital of Philadelphia, Abramson Research Building, Philadelphia, PA, USA #These authors contributed equally to this work Purpose: To investigate the anchoring of plasmid DNA/anti-DNA antibody/cationic lipid tri-complex (DAC micelles onto bisphosphonate-modified 316 L coronary stents for cardiovascular site-specific gene delivery. Methods: Stents were first modified with polyallylamine bisphosphonate (PAA-BP, thereby enabling the retention of a PAA-BP molecular monolayer that permits the anchoring (via vector-binding molecules of DAC micelles. DAC micelles were then chemically linked onto the PAA-BP-modified stents by using N-succinimidyl-3-(2-pyridyldithiol-propionate (SPDP as a crosslinker. Rhodamine-labeled DNA was used to assess the anchoring of DAC micelles, and radioactive-labeled antibody was used to evaluate binding capacity and stability. DAC micelles (encoding green fluorescent protein were tethered onto the PAA-BP-modified stents, which were assessed in cell culture. The presence of a PAA-BP molecular monolayer on the steel surface was confirmed by X-ray photoelectron spectroscopy and atomic force microscope analysis. Results: The anchoring of DAC micelles was generally uniform and devoid of large-scale patches of defects. Isotopic quantification confirmed that the amount of antibody chemically linked on the stents was 17-fold higher than that of the physical adsorbed control stents and its retention time was also significantly longer. In cell culture, numerous green fluorescent protein-positive cells were found on the PAA-BP modified stents, which demonstrated high localization and efficiency of gene delivery. Conclusion: The DAC micelle

  15. Loss of cell-surface laminin anchoring promotes tumor growth and is associated with poor clinical outcomes.

    Science.gov (United States)

    Akhavan, Armin; Griffith, Obi L; Soroceanu, Liliana; Leonoudakis, Dmitri; Luciani-Torres, Maria Gloria; Daemen, Anneleen; Gray, Joe W; Muschler, John L

    2012-05-15

    Perturbations in the composition and assembly of extracellular matrices (ECM) contribute to progression of numerous diseases, including cancers. Anchoring of laminins at the cell surface enables assembly and signaling of many ECMs, but the possible contributions of altered laminin anchoring to cancer progression remain undetermined. In this study, we investigated the prominence and origins of defective laminin anchoring in cancer cells and its association with cancer subtypes and clinical outcomes. We found loss of laminin anchoring to be widespread in cancer cells. Perturbation of laminin anchoring originated from several distinct defects, which all led to dysfunctional glycosylation of the ECM receptor dystroglycan. In aggressive breast and brain cancers, defective laminin anchoring was often due to suppressed expression of the glycosyltransferase LARGE. Reduced expression of LARGE characterized a broad array of human tumors in which it was associated with aggressive cancer subtypes and poor clinical outcomes. Notably, this defect robustly predicted poor survival in patients with brain cancers. Restoring LARGE expression repaired anchoring of exogenous and endogenous laminin and modulated cell proliferation and tumor growth. Together, our findings suggest that defects in laminin anchoring occur commonly in cancer cells, are characteristic of aggressive cancer subtypes, and are important drivers of disease progression.

  16. Study on mechanical principle of reinforcing sidewalls and corners of roadway to control floor heave by anchors

    Institute of Scientific and Technical Information of China (English)

    王卫军

    2003-01-01

    The movement principle of sidewalls and floor of extraction opening is analyzed, it is found that floor heave not only has something to do with the floor strata, but sidewalls. The effect of sidewall anchor and corner anchor is studied, the mechanical principle of reinforcing sidewalls and corners is put forward and applied in engineering practice.

  17. 15 CFR Appendix C to Subpart H of... - No-Anchoring Seagrass Protection Zones in Tomales Bay

    Science.gov (United States)

    2010-01-01

    ... 15 Commerce and Foreign Trade 3 2010-01-01 2010-01-01 false No-Anchoring Seagrass Protection Zones in Tomales Bay C Appendix C to Subpart H of Part 922 Commerce and Foreign Trade Regulations Relating... 922—No-Anchoring Seagrass Protection Zones in Tomales Bay Coordinates listed in this Appendix...

  18. Cell wall anchoring of the Campylobacter antigens to Lactococcus lactis

    Directory of Open Access Journals (Sweden)

    Patrycja Anna Kobierecka

    2016-02-01

    Full Text Available Campylobacter jejuni is the most frequent cause of human food-borne gastroenteritis and chicken meat is the main source of infection. Recent studies showed that broiler chicken immunization against Campylobacter should be the most efficient way to lower the number of human infections by this pathogen. Induction of the mucosal immune system after oral antigen administration should provide protective immunity to chickens. In this work we tested the usefulness of Lactococcus lactis, the most extensively studied lactic acid bacterium, as a delivery vector for Campylobacter antigens. First we constructed hybrid protein – CjaA antigen presenting CjaD peptide epitopes on its surface. We showed that specific rabbit anti-rCjaAD serum reacted strongly with both CjaA and CjaD produced by a wild type Campylobacter jejuni strain. Next, rCjaAD and CjaA were fused to the C-terminus of the L. lactis YndF containing the LPTXG motif. The genes expressing these proteins were transcribed under control of the L. lactis Usp45 promoter and their products contain the Usp45 signal sequences. This strategy ensures a cell surface location of both analysed proteins, which was confirmed by immunofluorescence assay. In order to evaluate the impact of antigen location on vaccine prototype efficacy, a L. lactis strain producing cytoplasm-located rCjaAD was also generated. Animal experiments showed a decrease of Campylobacter cecal load in vaccinated birds as compared with the control group and showed that the L. lactis harboring the surface-exposed rCjaAD antigen afforded greater protection than the L. lactis producing cytoplasm-located rCjaAD. To the best of our knowledge, this is the first attempt to employ LAB (Lactic Acid Bacteria strains as a mucosal delivery vehicle for chicken immunization. Although the observed reduction of chicken colonization by Campylobacter resulting from vaccination was rather moderate, the experiments showed that LAB strains can be considered

  19. Side-chain-anchored N(alpha)-Fmoc-Tyr-OPfp for bidirectional solid-phase synthesis

    DEFF Research Database (Denmark)

    Olsen, Christian A; Jørgensen, Malene; Hansen, Steen H;

    2005-01-01

    [reaction: see text] A mild resin-immobilization strategy employing a readily prepared trityl bromide resin for anchoring building blocks via a phenol group has been developed. With N(alpha)-Fmoc-Tyr-OPfp as a starter building block, it was possible to prepare asymmetrically substituted hybrids o...... of spider- and wasp-type polyamine toxins using solid-phase peptide synthesis conditions....

  20. Hearing rehabilitation in Treacher Collins Syndrome with bone anchored hearing aid

    Directory of Open Access Journals (Sweden)

    José Fernando Polanski

    2015-12-01

    Full Text Available Objective: To describe a case of hearing rehabilitation with bone anchored hearing aid in a patient with Treacher Collins syndrome. Case description: 3 years old patient, male, with Treacher Collins syndrome and severe complications due to the syndrome, mostly related to the upper airway and hearing. He had bilateral atresia of external auditory canals, and malformation of the pinna. The initial hearing rehabilitation was with bone vibration arch, but there was poor acceptance due the discomfort caused by skull compression. It was prescribed a model of bone-anchored hearing aid, in soft band format. The results were evaluated through behavioral hearing tests and questionnaires Meaningful Use of Speech Scale (MUSS and Infant-Toddler Meaningful Auditory Integration Scale (IT-MAIS. Comments: The patient had a higher acceptance of the bone-anchored hearing aid compared to the traditional bone vibration arch. Audiological tests and the speech and auditory skills assessments also showed better communication and hearing outcomes. The bone-anchored hearing aid is a good option in hearing rehabilitation in this syndrome.

  1. Hearing rehabilitation in Treacher Collins Syndrome with bone anchored hearing aid

    Science.gov (United States)

    Polanski, José Fernando; Plawiak, Anna Clara; Ribas, Angela

    2015-01-01

    Objective: To describe a case of hearing rehabilitation with bone anchored hearing aid in a patient with Treacher Collins syndrome. Case description: 3 years old patient, male, with Treacher Collins syndrome and severe complications due to the syndrome, mostly related to the upper airway and hearing. He had bilateral atresia of external auditory canals, and malformation of the pinna. The initial hearing rehabilitation was with bone vibration arch, but there was poor acceptance due the discomfort caused by skull compression. It was prescribed a model of bone-anchored hearing aid, in soft band format. The results were evaluated through behavioral hearing tests and questionnaires Meaningful Use of Speech Scale (MUSS) and Infant-Toddler Meaningful Auditory Integration Scale (IT-MAIS). Comments: The patient had a higher acceptance of the bone-anchored hearing aid compared to the traditional bone vibration arch. Audiological tests and the speech and auditory skills assessments also showed better communication and hearing outcomes. The bone-anchored hearing aid is a good option in hearing rehabilitation in this syndrome. PMID:26298651

  2. Parameter analysis of anchor bolt support for large-span and jointed rock mass

    Institute of Scientific and Technical Information of China (English)

    LI Xi-bing; ZHOU Zi-long; LI Qi-yue; HU Liu-qing

    2005-01-01

    In order to obtain the optimal parameters of anchor bolt supporting system for large-span and jointed rock mass in Kaiyang Phosphor Mine, it is expensive and unavailable with the method of in-situ experiments. This paper describes a numerical modeling with discrete element method for the supporting effects of different type of anchor bolts. The anchor bolts with variant length of 0.5 m, 0.8 m, 1.0 m, diameter of 10 mm, 15 mm, 20 mm, setting spacing of 3.0 m, 2.5 m, 2.0 m, and setting angle of 10°, 20°, 30°, are simulated respectively. The results show that there exist optimal parameters of anchor bolt support for large-span and jointed rock mass. For the bolt support of the concerning, the optimal length is 2.5-3.5 m, the diameter is 25-35 mm, the spacing is 0.5-0.6 m, and the setting angle is 105°.

  3. Effects of Thinking Style and Spatial Ability on Anchoring Behavior in Geographic Information Systems

    Science.gov (United States)

    Wang, Dai-Yi; Lee, Mei-Hsuan; Sun, Chuen-Tsai

    2013-01-01

    The authors propose an instructional use for Google Earth (a GIS application) as an anchoring tool for knowledge integration. Google Earth can be used to support student explorations of world geography based on Wikipedia articles on earth science and history topics. We asked 66 Taiwanese high-school freshmen to make place marks with explanatory…

  4. 48 CFR 225.7007 - Restrictions on anchor and mooring chain.

    Science.gov (United States)

    2010-10-01

    ... 48 Federal Acquisition Regulations System 3 2010-10-01 2010-10-01 false Restrictions on anchor and mooring chain. 225.7007 Section 225.7007 Federal Acquisition Regulations System DEFENSE ACQUISITION REGULATIONS SYSTEM, DEPARTMENT OF DEFENSE SOCIOECONOMIC PROGRAMS FOREIGN ACQUISITION Authorization...

  5. 48 CFR 252.225-7019 - Restriction on acquisition of anchor and mooring chain.

    Science.gov (United States)

    2010-10-01

    ... 48 Federal Acquisition Regulations System 3 2010-10-01 2010-10-01 false Restriction on acquisition of anchor and mooring chain. 252.225-7019 Section 252.225-7019 Federal Acquisition Regulations System DEFENSE ACQUISITION REGULATIONS SYSTEM, DEPARTMENT OF DEFENSE CLAUSES AND FORMS SOLICITATION...

  6. Physical and numerical modelling of earth pressure on anchored sheet pile walls in sand

    DEFF Research Database (Denmark)

    Krogsbøll, Anette Susanne; Fuglsang, Leif D

    2006-01-01

    The influence of wall flexibility on earth pressure, bending moments and failure modes is studied. Numerical models are compared to results from model tests carried out in a geotechnical centrifuge. The back-fill is dry sand and failure is introduced by allowing the wall to rotate around the anchor...

  7. Physical and numerical modelling of earth pressure on anchored sheet pile walls in sand

    DEFF Research Database (Denmark)

    Krogsbøll, Anette Susanne; Fuglsang, Leif D

    The influence of wall flexibility on earth pressure, bending moments and failure modes is studied. Numerical models are compared to results from model tests carried out in a geotechnical centrifuge. The back-fill is dry sand and failure is introduced by allowing the wall to rotate around the anchor...

  8. Arrows as Anchors: An Analysis of the Material Features of Electric Field Vector Arrows

    Science.gov (United States)

    Gire, Elizabeth; Price, Edward

    2014-01-01

    Representations in physics possess both physical and conceptual aspects that are fundamentally intertwined and can interact to support or hinder sense making and computation. We use distributed cognition and the theory of conceptual blending with material anchors to interpret the roles of conceptual and material features of representations in…

  9. Auditory frequency discrimination in adult dyslexics: A test of the anchoring hypothesis

    NARCIS (Netherlands)

    Wijnen, F.N.K.; Kappers, A.M.L.; Vlutters, L.D.; Winkel, S.

    2012-01-01

    Purpose: A recent hypothesis ascribes dyslexia to a perceptual anchoring deficit. Supporting results have so far been obtained only in children with dyslexia and additional learning difficulties, but the hypothesis has been argued to apply to all individuals with dyslexia. Method: The authors measur

  10. The Relationship between Situated Cognition and Anchored Instruction: A Response to Tripp.

    Science.gov (United States)

    Moore, Joyce L.; And Others

    1994-01-01

    Discusses the relationship between situated cognition and anchored instruction in response to criticism of a previous article. Topics addressed include school versus nonschool settings; learning by observing models; "real-world" problem solving; transfer of training; and future work. (26 references) (LRW)

  11. Comparing General and Special Education Preservice Teachers' Test Performance Using Traditional and Anchored Instruction

    Science.gov (United States)

    Malone, D. Michael; Langone, John

    2005-01-01

    Data comparing the effects of traditional lectures and instruction paired with video anchors on test scores of general education and special education preservice teachers are presented. The sample in both the experimental and control groups included a mixture of preservice teachers including those beginning a general education early childhood…

  12. Using Anchored Instruction to Teach Preservice Teachers to Integrate Technology in the Curriculum

    Science.gov (United States)

    Kariuki, Mumbi; Duran, Mesut

    2004-01-01

    This case study addresses the use of the "anchored instruction approach" to restructure educational computing courses to enhance future teachers' learning of technology applications in the classroom. A cohort group of 22 preservice teachers from a typical teacher education institution in Southeastern Ohio was involved in the study. The…

  13. A Description of the Impact of Multimedia Anchored Instruction on Classroom Interactions.

    Science.gov (United States)

    Glaser, Candyce Williams; Rieth, Herbert J.; Kinzer, Charles K.; Colburn, Linda K.; Peter, Jeanne

    1999-01-01

    A study explored effects of a multimedia-based anchored instruction intervention on student/teacher interactions in a social studies classroom with 19 eighth graders, 9 with mild disabilities. Overall, instruction became more interactive as observational and interview data indicated a twofold increase in the number of daily student/teacher…

  14. Anchoring Problem-Solving and Computation Instruction in Context-Rich Learning Environments

    Science.gov (United States)

    Bottge, Brian A.; Rueda, Enrique; Grant, Timothy S.; Stephens, Ana C.; Laroque, Perry T.

    2010-01-01

    Middle school students with learning disabilities in math (MLD) used two versions of Enhanced Anchored Instruction (EAI). In one condition, students learned how to compute with fractions on an as-needed basis while they worked to solve the EAI problems. In the other condition, teachers used a computer-based instructional module in place of one of…

  15. Slipping Anchor? Testing the Vignettes Approach to Identification and Correction of Reporting Heterogeneity

    Science.gov (United States)

    d'Uva, Teresa Bago; Lindeboom, Maarten; O'Donnell, Owen; van Doorslaer, Eddy

    2011-01-01

    We propose tests of the two assumptions under which anchoring vignettes identify heterogeneity in reporting of categorical evaluations. Systematic variation in the perceived difference between any two vignette states is sufficient to reject "vignette equivalence." "Response consistency"--the respondent uses the same response…

  16. Investigating Repeater Effects on Chained Equipercentile Equating with Common Anchor Items

    Science.gov (United States)

    Kim, Sooyeon; Walker, Michael E.

    2012-01-01

    This study investigated the impact of repeat takers of a licensure test on the equating functions in the context of a nonequivalent groups with anchor test (NEAT) design. Examinees who had taken a new, to-be-equated form of the test were divided into three subgroups according to their previous testing experience: (a) repeaters who previously took…

  17. Solid-phase oligosaccharide synthesis with tris(alkoxy)benzyl amine (BAL) safety-catch anchoring

    DEFF Research Database (Denmark)

    Tolborg, Jakob Fjord; Jensen, Knud Jørgen

    2000-01-01

    A tris(alkoxy)benzylamine (BAL) handle strategy was developed for safety-catch anchoring of D-glucosamine derivatives in solid-phase synthesis of oligosaccharides; the linkage between the BAL handle and the amine proved stable to conc. TFA and Lewis acids, but after N-acylation the amide could...

  18. Arthroscopic double-row suture anchor fixation of minimally displaced greater tuberosity fractures.

    Science.gov (United States)

    Ji, Jong-Hun; Kim, Weon-Yoo; Ra, Ki-Hang

    2007-10-01

    In cases of displaced greater tuberosity fractures, treatments by arthroscopic-assisted reduction and percutaneous screw fixation have been reported. However, in cases in which there is a comminuted fracture or a minimally displaced fracture combined with concomitant lesions such as rotator cuff tear or labral pathology, it is difficult to reduce the fracture and to treat other pathologies by use of a percutaneous screw. Recently, many surgeons have used the double-row repair method in rotator cuff repair, which provides a tendon-bone interface better suited for biologic healing and restoring normal anatomy. In accordance with this method, we used the arthroscopic technique of double-row suture anchor fixation for a minimally displaced greater tuberosity fracture without additional incision. Initially, debridement was performed on the fracture surface by use of a shaver, and the medial-row anchor was inserted through the anterior portal or the intact cuff. Two lateral-row anchors were inserted just anterior and posterior to the lower margin of the fractured fragment under C-arm guidance. The medial-row sutures and lateral-row sutures were then placed. Arthroscopic double-row suture anchor fixation of a displaced greater tuberosity fracture restores the original footprint of the rotator cuff and normal tendon-bone interface of the displaced greater tuberosity fracture.

  19. Dynamic calculation model and seismic response for frame supporting structure with prestressed anchors

    Institute of Scientific and Technical Information of China (English)

    2010-01-01

    A dynamic calculation model of frame supporting structures with prestressed anchors for the slope stability is proposed. The frame and soil are closely contacted in the role of prestressed anchors and they cannot be separated along the whole slope. The lateral displacement of frame and soil is nearly in phase. The movement characteristic satisfies the theory of elastic foundation beam. The frame is treated with elastic foundation beam in this model. The influence of prestressed anchors is simplified as linear spring and damped system related with velocity. Under the condition of horizontal earthquake excitation, the equation of vibration response is established by using the model of dynamic Winkler beam and the analytical solutions are obtained for simple harmonic vibration. This method is applied to a case record for illustration of its capability, in order to verify the method, 3D nonlinear FEM (ADINA) is used to analyze the seismic performance of this case, the comparative results show that the design and the analysis are safe and credible by using the proposed method. The calculation model provides a new way for earthquake analysis and seismic design of slope stability supported by frame structure with prestressed anchors.

  20. Experiments with result diversity and entity ranking: Text, anchors, links, and Wikipedia

    NARCIS (Netherlands)

    Kaptein, R.; Koolen, M.; Kamps, J.

    2009-01-01

    In this paper, we document our efforts in participating to the TREC 2009 Entity Ranking and Web Tracks. We had multiple aims: For the Web Track’s Adhoc task we experiment with document text and anchor text representation, and the use of the link structure. For the Web Track’s Diversity task we exper