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Sample records for intermediate filament protein

  1. Molecular evolution of type VI intermediate filament proteins

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    Vincent Michel

    2007-09-01

    Full Text Available Abstract Background Tanabin, transitin and nestin are type VI intermediate filament (IF proteins that are developmentally regulated in frogs, birds and mammals, respectively. Tanabin is expressed in the growth cones of embryonic vertebrate neurons, whereas transitin and nestin are found in myogenic and neurogenic cells. Another type VI IF protein, synemin, is expressed in undifferentiated and mature muscle cells of birds and mammals. In addition to an IF-typical α-helical core domain, type VI IF proteins are characterized by a long C-terminal tail often containing distinct repeated motifs. The molecular evolution of type VI IF proteins remains poorly studied. Results To examine the evolutionary history of type VI IF proteins, sequence comparisons, BLAST searches, synteny studies and phylogenic analyses were performed. This study provides new evidence that tanabin, transitin and nestin are indeed orthologous type VI IF proteins. It demonstrates that tanabin, transitin and nestin genes share intron positions and sequence identities, have a similar chromosomal context and display closely related positions in phylogenic analyses. Despite this homology, fast evolution rates of their C-terminal extremity have caused the appearance of repeated motifs with distinct biological activities. In particular, our in silico and in vitro analyses of their tail domain have shown that (avian transitin, but not (mammalian nestin, contains a repeat domain displaying nucleotide hydrolysis activity. Conclusion These analyses of the evolutionary history of the IF proteins fit with a model in which type VI IFs form a branch distinct from NF proteins and are composed of two major proteins: synemin and nestin orthologs. Rapid evolution of the C-terminal extremity of nestin orthologs could be responsible for their divergent functions.

  2. Bacterial intermediate filaments

    DEFF Research Database (Denmark)

    Charbon, Godefroid; Cabeen, M.; Jacobs-Wagner, C.

    2009-01-01

    Crescentin, which is the founding member of a rapidly growing family of bacterial cytoskeletal proteins, was previously proposed to resemble eukaryotic intermediate filament (IF) proteins based on structural prediction and in vitro polymerization properties. Here, we demonstrate that crescentin...... also shares in vivo properties of assembly and dynamics with IF proteins by forming stable filamentous structures that continuously incorporate subunits along their length and that grow in a nonpolar fashion. De novo assembly of crescentin is biphasic and involves a cell size-dependent mechanism...... a new function for MreB and providing a parallel to the role of actin in IF assembly and organization in metazoan cells. Additionally, analysis of an MreB localization mutant suggests that cell wall insertion during cell elongation normally occurs along two helices of opposite handedness, each...

  3. Epithelial Intermediate Filaments: Guardians against Microbial Infection?

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    Florian Geisler

    2016-06-01

    Full Text Available Intermediate filaments are abundant cytoskeletal components of epithelial tissues. They have been implicated in overall stress protection. A hitherto poorly investigated area of research is the function of intermediate filaments as a barrier to microbial infection. This review summarizes the accumulating knowledge about this interaction. It first emphasizes the unique spatial organization of the keratin intermediate filament cytoskeleton in different epithelial tissues to protect the organism against microbial insults. We then present examples of direct interaction between viral, bacterial, and parasitic proteins and the intermediate filament system and describe how this affects the microbe-host interaction by modulating the epithelial cytoskeleton, the progression of infection, and host response. These observations not only provide novel insights into the dynamics and function of intermediate filaments but also indicate future avenues to combat microbial infection.

  4. Glial fibrillary acidic protein (GFAP) and the astrocyte intermediate filament system in diseases of the central nervous system

    NARCIS (Netherlands)

    Hol, Elly M; Pekny, Milos

    Glial fibrillary acidic protein (GFAP) is the hallmark intermediate filament (IF; also known as nanofilament) protein in astrocytes, a main type of glial cells in the central nervous system (CNS). Astrocytes have a range of control and homeostatic functions in health and disease. Astrocytes assume a

  5. Glial fibrillary acidic protein (GFAP) and the astrocyte intermediate filament system in diseases of the central nervous system

    NARCIS (Netherlands)

    Hol, E.M.; Pekny, M.

    2015-01-01

    Glial fibrillary acidic protein (GFAP) is the hallmark intermediate filament (IF; also known as nanofilament) protein in astrocytes, a main type of glial cells in the central nervous system (CNS). Astrocytes have a range of control and homeostatic functions in health and disease. Astrocytes assume a

  6. The intermediate filament network protein, vimentin, is required for parvoviral infection

    Energy Technology Data Exchange (ETDEWEB)

    Fay, Nikta; Panté, Nelly, E-mail: pante@zoology.ubc.ca

    2013-09-15

    Intermediate filaments (IFs) have recently been shown to serve novel roles during infection by many viruses. Here we have begun to study the role of IFs during the early steps of infection by the parvovirus minute virus of mice (MVM). We found that during early infection with MVM, after endosomal escape, the vimentin IF network was considerably altered, yielding collapsed immunofluorescence staining near the nuclear periphery. Furthermore, we found that vimentin plays an important role in the life cycle of MVM. The number of cells, which successfully replicated MVM, was reduced in infected cells in which the vimentin network was genetically or pharmacologically modified; viral endocytosis, however, remained unaltered. Perinuclear accumulation of MVM-containing vesicles was reduced in cells lacking vimentin. Our data suggests that vimentin is required for the MVM life cycle, presenting possibly a dual role: (1) following MVM escape from endosomes and (2) during endosomal trafficking of MVM. - Highlights: • MVM infection changes the distribution of the vimentin network to perinuclear regions. • Disrupting the vimentin network with acrylamide decreases MVM replication. • MVM replication is significantly reduced in vimentin-null cells. • Distribution of MVM-containing vesicles is affected in MVM infected vimentin-null cells.

  7. FSHD myoblasts fail to downregulate intermediate filament protein vimentin during myogenic differentiation.

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    Lipinski M.

    2011-10-01

    Full Text Available Facioscapulohumeral muscular dystrophy (FSHD is an autosomal dominant hereditary neuromuscular disorder. The clinical features of FSHD include weakness of the facial and shoulder girdle muscles followed by wasting of skeletal muscles of the pelvic girdle and lower extremities. Although FSHD myoblasts grown in vitro can be induced to differentiate into myotubes by serum starvation, the resulting FSHD myotubes have been shown previously to be morphologically abnormal. Aim. In order to find the cause of morphological anomalies of FSHD myotubes we compared in vitro myogenic differentiation of normal and FSHD myoblasts at the protein level. Methods. We induced myogenic differentiation of normal and FSHD myoblasts by serum starvation. We then compared protein extracts from proliferating myoblasts and differentiated myotubes using SDS-PAGE followed by mass spectrometry identification of differentially expressed proteins. Results. We demonstrated that the expression of vimentin was elevated at the protein and mRNA levels in FSHD myotubes as compared to normal myotubes. Conclusions. We demonstrate for the first time that in contrast to normal myoblasts, FSHD myoblasts fail to downregulate vimentin after induction of in vitro myogenic differentiation. We suggest that vimentin could be an easily detectable marker of FSHD myotubes

  8. Regulation of protein phosphorylation of the intermediate-sized filament vimentin in the ciliary epithelium of the mammalian eye

    International Nuclear Information System (INIS)

    Coca-Prados, M.

    1985-01-01

    The intermediate-sized filaments of vimentin-type (Mr = 57,000) have been identified biochemically and immunochemically as a major cytoskeleton component in the ciliary epithelium of the mammalian eye. When human or rabbit ciliary processes, or cultured ciliary epithelial-derived cells were incubated in serum-free medium containing [ 32 P]orthophosphate and any of the following agents: 1) beta-adrenergic agonists (isoproterenol or epinephrine), 2) direct activators of adenylate cyclase (cholera toxin or forskolin), 3) analogs of cyclic AMP (8-Br-cAMP), or 4) prostaglandin E1, the phosphorylation of vimentin was significantly enhanced. The maximal enhancement ranged, in vivo and in vitro, from about 3-fold in human to 5-fold in rabbit, with either 1 mM 8-Br-cAMP or 0.1 microM forskolin. Indirect immunofluorescence microscopy using a monoclonal antibody, anti-vimentin, allowed the localization of vimentin filaments in cultured ciliary epithelial cells. Treatment of these cells in culture with the catecholamine hormone, isoproterenol (1 microM), resulted in a profound reorganization of vimentin filaments. This may be correlated with the enhanced levels of phosphorylated vimentin observed upon increasing cellular cyclic AMP

  9. Regulation of protein phosphorylation of the intermediate-sized filament vimentin in the ciliary epithelium of the mammalian eye

    Energy Technology Data Exchange (ETDEWEB)

    Coca-Prados, M.

    1985-08-25

    The intermediate-sized filaments of vimentin-type (Mr = 57,000) have been identified biochemically and immunochemically as a major cytoskeleton component in the ciliary epithelium of the mammalian eye. When human or rabbit ciliary processes, or cultured ciliary epithelial-derived cells were incubated in serum-free medium containing (TSP)orthophosphate and any of the following agents: 1) beta-adrenergic agonists (isoproterenol or epinephrine), 2) direct activators of adenylate cyclase (cholera toxin or forskolin), 3) analogs of cyclic AMP (8-Br-cAMP), or 4) prostaglandin E1, the phosphorylation of vimentin was significantly enhanced. The maximal enhancement ranged, in vivo and in vitro, from about 3-fold in human to 5-fold in rabbit, with either 1 mM 8-Br-cAMP or 0.1 microM forskolin. Indirect immunofluorescence microscopy using a monoclonal antibody, anti-vimentin, allowed the localization of vimentin filaments in cultured ciliary epithelial cells. Treatment of these cells in culture with the catecholamine hormone, isoproterenol (1 microM), resulted in a profound reorganization of vimentin filaments. This may be correlated with the enhanced levels of phosphorylated vimentin observed upon increasing cellular cyclic AMP.

  10. Development of a Novel Green Fluorescent Protein-Based Binding Assay to Study the Association of Plakins with Intermediate Filament Proteins.

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    Favre, Bertrand; Begré, Nadja; Bouameur, Jamal-Eddine; Borradori, Luca

    2016-01-01

    Protein-protein interactions are fundamental for most biological processes, such as the formation of cellular structures and enzymatic complexes or in signaling pathways. The identification and characterization of protein-protein interactions are therefore essential for understanding the mechanisms and regulation of biological systems. The organization and dynamics of the cytoskeleton, as well as its anchorage to specific sites in the plasma membrane and organelles, are regulated by the plakins. These structurally related proteins anchor different cytoskeletal networks to each other and/or to other cellular structures. The association of several plakins with intermediate filaments (IFs) is critical for maintenance of the cytoarchitecture. Pathogenic mutations in the genes encoding different plakins can lead to dramatic manifestations, occurring principally in the skin, striated muscle, and/or nervous system, due to cytoskeletal disorganization resulting in abnormal cell fragility. Nevertheless, it is still unclear how plakins bind to IFs, although some general rules are slowly emerging. We here describe in detail a recently developed protein-protein fluorescence binding assay, based on the production of recombinant proteins tagged with green fluorescent protein (GFP) and their use as fluid-phase fluorescent ligands on immobilized IF proteins. Using this method, we have been able to assess the ability of C-terminal regions of GFP-tagged plakin proteins to bind to distinct IF proteins and IF domains. This simple and sensitive technique, which is expected to facilitate further studies in this area, can also be potentially employed for any kind of protein-protein interaction studies. © 2016 Elsevier Inc. All rights reserved.

  11. Unraveling Intermediate Filaments : The super resolution solution

    NARCIS (Netherlands)

    Nahidiazar, L.

    2017-01-01

    Intermediate Filaments (IFs) carry out major functions in cells. Several diseases have been associated with malfunctioning IFs in the cells and among them are certain sub types of cancer. To determine the structure and organization of IFs, we have used Single Molecule Localization Microscopy (SMLM)

  12. Nonlinear Loading-Rate-Dependent Force Response of Individual Vimentin Intermediate Filaments to Applied Strain

    NARCIS (Netherlands)

    Block, Johanna; Witt, Hannes; Candelli, Andrea; Peterman, Erwin J. G.; Wuite, Gijs J. L.; Janshoff, Andreas; Koester, Sarah

    2017-01-01

    The mechanical properties of eukaryotic cells are to a great extent determined by the cytoskeleton, a composite network of different filamentous proteins. Among these, intermediate filaments (IFs) are exceptional in their molecular architecture and mechanical properties. Here we directly record

  13. Immunocytochemical demonstration of intermediate filament proteins, S-100 protein and CEA in apocrine sweat glands and apocrine gland derived lesions of the dog.

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    Ferrer, L; Rabanal, R M; Fondevila, D; Prats, N

    1990-09-01

    The presence of carcinoembryonic antigen (CEA), intermediate filament proteins and S-100 protein in normal and pathological canine apocrine sweat glands was investigated, using a standard immunoperoxidase technique. The normal apocrine sweat glands showed a constant immunoreactivity in all the cases studied. The cells of the acini and of the ducts only reacted with the antikeratin antibody. The myoepithelial cells reacted positively with the antisera antikeratin and anti protein S-100. Epithelial cells of apocrine cysts, sweat gland adenomas and sweat gland carcinomas showed the same immunoreaction than normal apocrine cells. Proliferating myoepithelial cells were also positive for vimentin. In two out of three adenocarcinomas a positive reaction with the anti CEA could be detected in the glandular cells. This can be due to the presence in glandular cells of CEA or of Nonspecific Crossreacting Antigen (NCA). These findings indicate that demonstration of keratin is a useful aid in the detection of apocrine gland derived lesions in the dog. Similarly, S-100 protein is a marker for myoepithelial cells. Further research is necessary to investigate the expression of CEA in canine tumours.

  14. Primitive neuroectodermal tumors of the central nervous system. Patterns of expression of neuroendocrine markers, and all classes of intermediate filament proteins.

    NARCIS (Netherlands)

    Gould, V E; Jansson, D S; Molenaar, W M; Rorke, L B; Trojanowski, J Q; Lee, V M; Packer, R J; Franke, W W

    1990-01-01

    Snap-frozen samples from 22 primitive neuroectodermal tumors (PNETs) primary in the central nervous system were studied with antibodies to synaptophysin, bombesin, somatostatin, substance P, vasoactive intestinal polypeptide, all classes of intermediate filaments, and desmoplakins I and II. Frozen

  15. Intermediate filament mechanics in vitro and in the cell: From coiled coils to filaments, fibers and networks

    OpenAIRE

    Köster, Sarah; Weitz, David; Goldman, Robert D.; Aebi, Ueli; Herrmann, Harald

    2015-01-01

    Intermediate filament proteins form filaments, fibers and networks both in the cytoplasm and the nucleus of metazoan cells. Their general structural building plan accommodates highly varying amino acid sequences to yield extended dimeric α-helical coiled coils of highly conserved design. These “rod” particles are the basic building blocks of intrinsically flexible, filamentous structures that are able to resist high mechanical stresses, i.e. bending and stretching to a considerable degree, bo...

  16. Assembly studies of six intestinal intermediate filament (IF) proteins B2, C1, C2, D1, D2, and E1 in the nematode C. elegans.

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    Karabinos, Anton; Schünemann, Jürgen; Parry, David A D

    2017-03-01

    The dimerisation properties of six intestine-expressed intermediate filament (IF) proteins (B2, C1, C2, D1, D2, E1) were analysed in blot overlay assay on membranes containing all of the eleven recombinant C. elegans IF proteins (A1, A2, A3, A4, B1, B2, C1, C2, D1, D2, and E1). The interactions detected in the blot assays exclusively comprise intestine-expressed IF proteins and the protein A4, which is found in the dauer larva intestine. About 86% of these interactions are heterotypic, while the remaining interactions relate to C1, C2, and D2 homodimers. These multiple modes of interaction were also supported by calculations of the numbers of possible interchain ionic interactions derived from the individual rod sequences. The results predict that the six B2, C1, C2, D1, D2, and E1 IF proteins are able to form as many as eleven different heteropolymeric and three homopolymeric IFs in the C. elegans intestine. This simple model of the intestinal IF meshwork enables us to speculate that our previously reported triple RNAi worms arrested or decreased their growth because of feeding reduction due to morphological defects of the mechanically compromised intestine. © 2017 Wiley Periodicals, Inc.

  17. Attractive interactions among intermediate filaments determine network mechanics in vitro.

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    Paul Pawelzyk

    Full Text Available Mechanical and structural properties of K8/K18 and vimentin intermediate filament (IF networks have been investigated using bulk mechanical rheometry and optical microrheology including diffusing wave spectroscopy and multiple particle tracking. A high elastic modulus G0 at low protein concentration c, a weak concentration dependency of G0 (G0 ∼ c(0.5 ± 0.1 and pronounced strain stiffening are found for these systems even without external crossbridgers. Strong attractive interactions among filaments are required to maintain these characteristic mechanical features, which have also been reported for various other IF networks. Filament assembly, the persistence length of the filaments and the network mesh size remain essentially unaffected when a nonionic surfactant is added, but strain stiffening is completely suppressed, G0 drops by orders of magnitude and exhibits a scaling G0 ∼ c(1.9 ± 0.2 in agreement with microrheological measurements and as expected for entangled networks of semi-flexible polymers. Tailless K8Δ/K18ΔT and various other tailless filament networks do not exhibit strain stiffening, but still show high G0 values. Therefore, two binding sites are proposed to exist in IF networks. A weaker one mediated by hydrophobic amino acid clusters in the central rod prevents stretched filaments between adjacent cross-links from thermal equilibration and thus provides the high G0 values. Another strong one facilitating strain stiffening is located in the tail domain with its high fraction of hydrophobic amino acid sequences. Strain stiffening is less pronounced for vimentin than for K8/K18 due to electrostatic repulsion forces partly compensating the strong attraction at filament contact points.

  18. Covisualization in living onion cells of putative integrin, putative spectrin, actin, putative intermediate filaments, and other proteins at the cell membrane and in an endomembrane sheath

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    Reuzeau, C.; Doolittle, K. W.; McNally, J. G.; Pickard, B. G.; Evans, M. L. (Principal Investigator)

    1997-01-01

    Covisualizations with wide-field computational optical-sectioning microscopy of living epidermal cells of the onion bulb scale have evidenced two major new cellular features. First, a sheath of cytoskeletal elements clads the endomembrane system. Similar elements clad the inner faces of punctate plasmalemmal sites interpreted as plasmalemmal control centers. One component of the endomembrane sheath and plasmalemmal control center cladding is anti-genicity-recognized by two injected antibodies against animal spectrin. Immunoblots of separated epidermal protein also showed bands recognized by these antibodies. Injected phalloidin identified F-actin with the same cellular distribution pattern, as did antibodies against intermediate-filament protein and other cytoskeletal elements known from animal cells. Injection of general protein stains demonstrated the abundance of endomembrane sheath protein. Second, the endomembrane system, like the plasmalemmal puncta, contains antigen recognized by an anti-beta 1 integrin injected into the cytoplasm. Previously, immunoblots of separated epidermal protein were shown to have a major band recognized both by this antibody prepared against a peptide representing the cytosolic region of beta 1 integrin and an antibody against the matrix region of beta 1 integrin. The latter antiboby also identified puncta at the external face of protoplasts. It is proposed that integrin and associated transmembrane proteins secure the endomembrane sheath and transmit signals between it and the lumen or matrix of the endoplasmic reticulum and organellar matrices. This function is comparable to that proposed for such transmembrane linkers in the plasmalemmal control centers, which also appear to bind cytoskeleton and a host of related molecules and transmit signals between them and the wall matrix. It is at the plasmalemmal control centers that the endoplasmic reticulum, a major component of the endomembrane system, attaches to the plasma membrane.

  19. Caveolae in fibroblast-like synoviocytes: static structures associated with vimentin-based intermediate filaments

    DEFF Research Database (Denmark)

    Berg, Kasper; Tamas, Raluca; Riemann, Anne

    2008-01-01

    . Vimentin was identified as the most abundant protein in detergent resistant membranes (DRM's), and by immunogold electron microscopy caveolae were seen in intimate contact with intermediate-size filaments. These observations indicate that vimentin-based filaments are responsible for the spatio...

  20. Chirality of Intermediate Filaments and Magnetic Helicity of Active Regions

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    Lim, Eun-Kyung; Chae, J.

    2009-05-01

    Filaments that form either between or around active regions (ARs) are called intermediate filaments. Even though there have been many theoretical studies, the origin of the chirality of filaments is still unknown. We investigated how intermediate filaments are related to their associated ARs, especially from the point of view of magnetic helicity and the orientation of polarity inversion lines (PILs). The chirality of filaments has been determined based on the orientations of barbs observed in the full-disk Hα images taken at Big Bear Solar Observatory during the rising phase of solar cycle 23. The sign of magnetic helicity of ARs has been determined using S/inverse-S shaped sigmoids from Yohkoh SXT images. As a result, we have found a good correlation between the chirality of filaments and the magnetic helicity sign of ARs. Among 45 filaments, 42 filaments have shown the same sign as helicity sign of nearby ARs. It has been also confirmed that the role of both the orientation and the relative direction of PILs to ARs in determining the chirality of filaments is not significant, against a theoretical prediction. These results suggest that the chirality of intermediate filaments may originate from magnetic helicity of their associated ARs.

  1. The calcium-modulated proteins, S100A1 and S100B, as potential regulators of the dynamics of type III intermediate filaments

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    M. Garbuglia

    1999-10-01

    Full Text Available The Ca2+-modulated, dimeric proteins of the EF-hand (helix-loop-helix type, S100A1 and S100B, that have been shown to inhibit microtubule (MT protein assembly and to promote MT disassembly, interact with the type III intermediate filament (IF subunits, desmin and glial fibrillary acidic protein (GFAP, with a stoichiometry of 2 mol of IF subunit/mol of S100A1 or S100B dimer and an affinity of 0.5-1.0 µM in the presence of a few micromolar concentrations of Ca2+. Binding of S100A1 and S100B results in inhibition of desmin and GFAP assemblies into IFs and stimulation of the disassembly of preformed desmin and GFAP IFs. S100A1 and S100B interact with a stretch of residues in the N-terminal (head domain of desmin and GFAP, thereby blocking the head-to-tail process of IF elongation. The C-terminal extension of S100A1 (and, likely, S100B represents a critical part of the site that recognizes desmin and GFAP. S100B is localized to IFs within cells, suggesting that it might have a role in remodeling IFs upon elevation of cytosolic Ca2+ concentration by avoiding excess IF assembly and/or promoting IF disassembly in vivo. S100A1, that is not localized to IFs, might also play a role in the regulation of IF dynamics by binding to and sequestering unassembled IF subunits. Together, these observations suggest that S100A1 and S100B may be regarded as Ca2+-dependent regulators of the state of assembly of two important elements of the cytoskeleton, IFs and MTs, and, potentially, of MT- and IF-based activities.

  2. Quantum dot labeling and imaging of glial fibrillary acidic protein intermediate filaments and gliosis in the rat neural retina and dissociated astrocytes.

    Science.gov (United States)

    Pathak, Smita; Tolentino, Rosa; Nguyen, Kim; D'Amico, Lorenzo; Barron, Erin; Cheng, Lingyun; Freeman, William R; Silva, Gabriel A

    2009-08-01

    The use of antibody and peptide functionalized semiconductor quantum dots holds considerable potential for specific labeling of target antigens and high resolution optical imaging of biological preparations. Despite this potential, their use in neuroscience is not yet widespread; a number of technical and methodological challenges must still be overcome in order to produce reliable and reproducible labeling protocols. We have optimized and used anti-GFAP functionalized quantum dots for specific labeling of intermediate filaments in astrocyte and Müller glial cells in sections of intact rat neural sensory retina and dissociated primary spinal cord astrocytes. These techniques produced stable and robust imaging of retinal astrocytes and Müller cells with minimal non-specific background labeling and intense fluorescence resulting in a high signal to noise ratio. This resulted in clear and efficient labeling of normal levels of GFAP in the retina and the ability to differentiate it from pathologically high levels of GFAP associated with reactive gliosis in a laser induced injury model. Labeling and imaging of dissociated astrocytes demonstrated the presence of what appeared to be highly complex organizations of fine intermediate filaments that spanned between cells to form intricate networks of filamentous intercellular bridges. The presence of these structures in situ and in vivo as well as any potential functions remain to be determined, but their identification should be greatly facilitated by quantum dot labeling protocols.

  3. Herpesviruses and Intermediate Filaments: Close Encounters with the Third Type

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    Laura Hertel

    2011-07-01

    Full Text Available Intermediate filaments (IF are essential to maintain cellular and nuclear integrity and shape, to manage organelle distribution and motility, to control the trafficking and pH of intracellular vesicles, to prevent stress-induced cell death, and to support the correct distribution of specific proteins. Because of this, IF are likely to be targeted by a variety of pathogens, and may act in favor or against infection progress. As many IF functions remain to be identified, however, little is currently known about these interactions. Herpesviruses can infect a wide variety of cell types, and are thus bound to encounter the different types of IF expressed in each tissue. The analysis of these interrelationships can yield precious insights into how IF proteins work, and into how viruses have evolved to exploit these functions. These interactions, either known or potential, will be the focus of this review.

  4. Phosphorylation and disassembly of intermediate filaments in mitotic cells

    International Nuclear Information System (INIS)

    Chou, Yinghao; Rosevear, E.; Goldman, R.D.

    1989-01-01

    As baby hamster kidney (BHK-21) cells enter mitosis, networks of intermediate filaments (IFs) are transformed into cytoplasmic aggregates of protofilaments. Coincident with this morphological change, the phosphate content of vimentin increases from 0.3 mol of P i per mol of protein in interphase to 1.9 mol of P i per mol of protein in mitosis. A similar increase in phosphate content is observed with desmin, from 0.5 mol of P i per mol of protein to 1.5 mol of P i per mol of protein. Fractionation of mitotic cell lysates by hydroxylapatite column chromatography reveals the presence of two IF protein kinase activities, designated as IF protein kinase I and IF protein kinase II. Comparison of two-dimensional 32 P-labeled phosphopeptide maps of vimentin and desmin phosphorylated in vivo in mitosis, and in vitro using partially purified kinase fractions, reveals extensive similarity in the two sets of phosphorylation sites. Phosphorylation of in vitro polymerized IFs by IF protein kinase II induces complete disassembly as determined by negative-stain electron microscopy. The results support the idea that the disassembly of IFs in mitosis is regulated by the phosphorylation of its subunit proteins

  5. Biological adhesion of the flatworm Macrostomum lignano relies on a duo-gland system and is mediated by a cell type-specific intermediate filament protein.

    Science.gov (United States)

    Lengerer, Birgit; Pjeta, Robert; Wunderer, Julia; Rodrigues, Marcelo; Arbore, Roberto; Schärer, Lukas; Berezikov, Eugene; Hess, Michael W; Pfaller, Kristian; Egger, Bernhard; Obwegeser, Sabrina; Salvenmoser, Willi; Ladurner, Peter

    2014-02-12

    Free-living flatworms, in both marine and freshwater environments, are able to adhere to and release from a substrate several times within a second. This reversible adhesion relies on adhesive organs comprised of three cell types: an adhesive gland cell, a releasing gland cell, and an anchor cell, which is a modified epidermal cell responsible for structural support. However, nothing is currently known about the molecules that are involved in this adhesion process. In this study we present the detailed morphology of the adhesive organs of the free-living marine flatworm Macrostomum lignano. About 130 adhesive organs are located in a horse-shoe-shaped arc along the ventral side of the tail plate. Each organ consists of exactly three cells, an adhesive gland cell, a releasing gland cell, and an anchor cell. The necks of the two gland cells penetrate the anchor cell through a common pore. Modified microvilli of the anchor cell form a collar surrounding the necks of the adhesive- and releasing glands, jointly forming the papilla, the outer visible part of the adhesive organs. Next, we identified an intermediate filament (IF) gene, macif1, which is expressed in the anchor cells. RNA interference mediated knock-down resulted in the first experimentally induced non-adhesion phenotype in any marine animal. Specifically, the absence of intermediate filaments in the anchor cells led to papillae with open tips, a reduction of the cytoskeleton network, a decline in hemidesmosomal connections, and to shortened microvilli containing less actin. Our findings reveal an elaborate biological adhesion system in a free-living flatworm, which permits impressively rapid temporary adhesion-release performance in the marine environment. We demonstrate that the structural integrity of the supportive cell, the anchor cell, is essential for this adhesion process: the knock-down of the anchor cell-specific intermediate filament gene resulted in the inability of the animals to adhere. The RNAi

  6. Withaferin a alters intermediate filament organization, cell shape and behavior.

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    Boris Grin

    Full Text Available Withaferin A (WFA is a steroidal lactone present in Withania somnifera which has been shown in vitro to bind to the intermediate filament protein, vimentin. Based upon its affinity for vimentin, it has been proposed that WFA can be used as an anti-tumor agent to target metastatic cells which up-regulate vimentin expression. We show that WFA treatment of human fibroblasts rapidly reorganizes vimentin intermediate filaments (VIF into a perinuclear aggregate. This reorganization is dose dependent and is accompanied by a change in cell shape, decreased motility and an increase in vimentin phosphorylation at serine-38. Furthermore, vimentin lacking cysteine-328, the proposed WFA binding site, remains sensitive to WFA demonstrating that this site is not required for its cellular effects. Using analytical ultracentrifugation, viscometry, electron microscopy and sedimentation assays we show that WFA has no effect on VIF assembly in vitro. Furthermore, WFA is not specific for vimentin as it disrupts the cellular organization and induces perinuclear aggregates of several other IF networks comprised of peripherin, neurofilament-triplet protein, and keratin. In cells co-expressing keratin IF and VIF, the former are significantly less sensitive to WFA with respect to inducing perinuclear aggregates. The organization of microtubules and actin/microfilaments is also affected by WFA. Microtubules become wavier and sparser and the number of stress fibers appears to increase. Following 24 hrs of exposure to doses of WFA that alter VIF organization and motility, cells undergo apoptosis. Lower doses of the drug do not kill cells but cause them to senesce. In light of our findings that WFA affects multiple IF systems, which are expressed in many tissues of the body, caution is warranted in its use as an anti-cancer agent, since it may have debilitating organism-wide effects.

  7. Unconventional actin conformations localize on intermediate filaments in mitosis

    International Nuclear Information System (INIS)

    Hubert, Thomas; Vandekerckhove, Joel; Gettemans, Jan

    2011-01-01

    Research highlights: → Unconventional actin conformations colocalize with vimentin on a cage-like structure in metaphase HEK 293T cells. → These conformations are detected with the anti-actin antibodies 1C7 ('lower dimer') and 2G2 ('nuclear actin'), but not C4 (monomeric actin). → Mitotic unconventional actin cables are independent of filamentous actin or microtubules. → Unconventional actin colocalizes with vimentin on a nocodazole-induced perinuclear dense mass of cables. -- Abstract: Different structural conformations of actin have been identified in cells and shown to reside in distinct subcellular locations of cells. In this report, we describe the localization of actin on a cage-like structure in metaphase HEK 293T cells. Actin was detected with the anti-actin antibodies 1C7 and 2G2, but not with the anti-actin antibody C4. Actin contained in this structure is independent of microtubules and actin filaments, and colocalizes with vimentin. Taking advantage of intermediate filament collapse into a perinuclear dense mass of cables when microtubules are depolymerized, we were able to relocalize actin to such structures. We hypothesize that phosphorylation of intermediate filaments at mitosis entry triggers the recruitment of different actin conformations to mitotic intermediate filaments. Storage and partition of the nuclear actin and antiparallel 'lower dimer' actin conformations between daughter cells possibly contribute to gene transcription and transient actin filament dynamics at G1 entry.

  8. Cornification in reptilian epidermis occurs through the deposition of keratin-associated beta-proteins (beta-keratins) onto a scaffold of intermediate filament keratins.

    Science.gov (United States)

    Alibardi, Lorenzo

    2013-02-01

    The isolation of genes for alpha-keratins and keratin-associated beta-proteins (formerly beta-keratins) has allowed the production of epitope-specific antibodies for localizing these proteins during the process of cornification epidermis of reptilian sauropsids. The antibodies are directed toward proteins in the alpha-keratin range (40-70 kDa) or beta-protein range (10-30 kDa) of most reptilian sauropsids. The ultrastructural immunogold study shows the localization of acidic alpha-proteins in suprabasal and precorneous epidermal layers in lizard, snake, tuatara, crocodile, and turtle while keratin-associated beta-proteins are localized in precorneous and corneous layers. This late activation of the synthesis of keratin-associated beta-proteins is typical for keratin-associated and corneous proteins in mammalian epidermis (involucrin, filaggrin, loricrin) or hair (tyrosine-rich or sulfur-rich proteins). In turtles and crocodilians epidermis, keratin-associated beta-proteins are synthesized in upper spinosus and precorneous layers and accumulate in the corneous layer. The complex stratification of lepidosaurian epidermis derives from the deposition of specific glycine-rich versus cysteine-glycine-rich keratin-associated beta-proteins in cells sequentially produced from the basal layer and not from the alternation of beta- with alpha-keratins. The process gives rise to Oberhäutchen, beta-, mesos-, and alpha-layers during the shedding cycle of lizards and snakes. Differently from fish, amphibian, and mammalian keratin-associated proteins (KAPs) of the epidermis, the keratin-associated beta-proteins of sauropsids are capable to form filaments of 3-4 nm which give rise to an X-ray beta-pattern as a consequence of the presence of a beta-pleated central region of high homology, which seems to be absent in KAPs of the other vertebrates. Copyright © 2012 Wiley Periodicals, Inc.

  9. The vital role of actin and the intermediate filaments in the ...

    African Journals Online (AJOL)

    The cytoskeleton is a system of structural protein primarily involved in the cellular architectural maintenance, intracellular transport, cell division and movement. The classical major components of the cytoskeleton are microtubules, intermediate filaments and microfilaments. Traditionally the microtubules are involved in ...

  10. The Autoimmune Regulator (AIRE), Which Is Defective in Autoimmune Polyendocrinopathy-Candidiasis-Ectodermal Dystrophy Patients, Is Expressed in Human Epidermal and Follicular Keratinocytes and Associates With the Intermediate Filament Protein Cytokeratin 17

    Science.gov (United States)

    Kumar, Vipul; Pedroza, Luis A.; Mace, Emily M.; Seeholzer, Steven; Cotsarelis, George; Condino-Neto, Antonio; Payne, Aimee S.; Orange, Jordan S.

    2011-01-01

    Autoimmune polyendocrinopathy-candidiasis-ectodermal dystrophy (APECED) syndrome, which is caused by mutation of the autoimmune regulator (AIRE) gene, is a highly variable disease characterized by multiple endocrine failure, chronic mucocutaneous candidiasis, and various ectodermal defects. AIRE is a transcriptional regulator classically expressed in medullary thymic epithelial cells, monocytes, macrophages, and dendritic cells. Previous studies have suggested that AIRE can shuttle between the nucleus and cytoplasm of cells, although its cytoplasmic functions are poorly characterized. Through mass spectrometry analysis of proteins co-immunoprecipitating with cytoplasmic AIRE, we identified a novel association of AIRE with the intermediate filament protein cytokeratin 17 (K17) in the THP-1 monocyte cell line. We confirmed AIRE expression in HaCaT epidermal keratinocytes, as well as its interaction with K17. Confocal microscopy of human fetal and adult scalp hair follicles demonstrated a cytoplasmic pattern of AIRE staining that moderately colocalized with K17. The cytoplasmic association of AIRE with the intermediate filament network in human epidermal and follicular keratinocytes may provide a new path to understanding the ectodermal abnormalities associated with the APECED syndrome. PMID:21356351

  11. Structural and Mechanical Properties of Intermediate Filaments under Extreme Conditions and Disease

    Science.gov (United States)

    Qin, Zhao

    Intermediate filaments are one of the three major components of the cytoskeleton in eukaryotic cells. It was discovered during the recent decades that intermediate filament proteins play key roles to reinforce cells subjected to large-deformation as well as participate in signal transduction. However, it is still poorly understood how the nanoscopic structure, as well as the biochemical properties of these protein molecules contribute to their biomechanical functions. In this research we investigate the material function of intermediate filaments under various extreme mechanical conditions as well as disease states. We use a full atomistic model and study its response to mechanical stresses. Learning from the mechanical response obtained from atomistic simulations, we build mesoscopic models following the finer-trains-coarser principles. By using this multiple-scale model, we present a detailed analysis of the mechanical properties and associated deformation mechanisms of intermediate filament network. We reveal the mechanism of a transition from alpha-helices to beta-sheets with subsequent intermolecular sliding under mechanical force, which has been inferred previously from experimental results. This nanoscale mechanism results in a characteristic nonlinear force-extension curve, which leads to a delocalization of mechanical energy and prevents catastrophic fracture. This explains how intermediate filament can withstand extreme mechanical deformation of > 1 00% strain despite the presence of structural defects. We combine computational and experimental techniques to investigate the molecular mechanism of Hutchinson-Gilford progeria syndrome, a premature aging disease. We find that the mutated lamin tail .domain is more compact and stable than the normal one. This altered structure and stability may enhance the association of intermediate filaments with the nuclear membrane, providing a molecular mechanism of the disease. We study the nuclear membrane association

  12. Nonlinear Loading-Rate-Dependent Force Response of Individual Vimentin Intermediate Filaments to Applied Strain

    Science.gov (United States)

    Block, Johanna; Witt, Hannes; Candelli, Andrea; Peterman, Erwin J. G.; Wuite, Gijs J. L.; Janshoff, Andreas; Köster, Sarah

    2017-01-01

    The mechanical properties of eukaryotic cells are to a great extent determined by the cytoskeleton, a composite network of different filamentous proteins. Among these, intermediate filaments (IFs) are exceptional in their molecular architecture and mechanical properties. Here we directly record stress-strain curves of individual vimentin IFs using optical traps and atomic force microscopy. We find a strong loading rate dependence of the mechanical response, supporting the hypothesis that IFs could serve to protect eukaryotic cells from fast, large deformations. Our experimental results show different unfolding regimes, which we can quantitatively reproduce by an elastically coupled system of multiple two-state elements.

  13. Complete Structure of an Epithelial Keratin Dimer: Implications for Intermediate Filament Assembly.

    Directory of Open Access Journals (Sweden)

    David J Bray

    Full Text Available Keratins are cytoskeletal proteins that hierarchically arrange into filaments, starting with the dimer sub-unit. They are integral to the structural support of cells, in skin, hair and nails. In skin, keratin is thought to play a critical role in conferring the barrier properties and elasticity of skin. In general, the keratin dimer is broadly described by a tri-domain structure: a head, a central rod and a tail. As yet, no atomistic-scale picture of the entire dimer structure exists; this information is pivotal for establishing molecular-level connections between structure and function in intermediate filament proteins. The roles of the head and tail domains in facilitating keratin filament assembly and function remain as open questions. To address these, we report results of molecular dynamics simulations of the entire epithelial human K1/K10 keratin dimer. Our findings comprise: (1 the first three-dimensional structural models of the complete dimer unit, comprising of the head, rod and tail domains; (2 new insights into the chirality of the rod-domain twist gained from analysis of the full domain structure; (3 evidence for tri-subdomain partitioning in the head and tail domains; and, (4 identification of the residue characteristics that mediate non-covalent contact between the chains in the dimer. Our findings are immediately applicable to other epithelial keratins, such as K8/K18 and K5/K14, and to intermediate filament proteins in general.

  14. Hyperthyreosis effects on the learning and glial intermediate filaments of rat brain

    Directory of Open Access Journals (Sweden)

    S. V. Kyrychenko

    2014-03-01

    Full Text Available The influence of hyperthyreosis on oxidative stress, state of glial intermediate filaments and memotry was investigated. Significant increasing of lipid peroxidation products into both hippocampus and cortex and change for the worse of memory was observed. Analysis of the behavioral reactions of rats in the test of passive avoidance conditioned reflex showed that the acquisition of skills of all groups of animals did not differ by time waiting period (latent period. Time saving memory test conditioned reflex of passive avoidance was excellent in the group of rats treated with thyroxine compared with controls. The change of polypeptide GFAP was observed in hippocampus and cortex. Both soluble and filamentous forms of GFAP increased in hippocampus of rat with hyperthyreosis. In filament fractions, increase in the intensity of 49 kDa polypeptide band was found. In the same fraction of insoluble cytoskeleton proteins degraded HFKB polypeptides with molecular weight in the region of 46–41 kDa appeared. Marked increase of degraded polypeptides was found in the soluble fraction of the brain stem. The intensity of the intact polypeptide (49 kDa, as well as in the filament fraction, significantly increased. It is possible that increasing concentrations of soluble subunits glial filaments may be due to dissociation of own filaments during the reorganization of cytoskeleton structures. Given the results of Western blotting for filament fraction, increased content of soluble intact 49 kDa polypeptide is primarily the result of increased expression of HFKB and only partly due to redistribution of existing filament structures. Calculation and analysis of indicators showed high correlation between the increase in content and peroxidation products of HFKB. These results indicate the important role of oxidative stress in the induction of astroglial reactive response under conditions of hyperthyroidism. This data shows the possibility of the glial cell

  15. Prokaryotic cytoskeletons: protein filaments organizing small cells.

    Science.gov (United States)

    Wagstaff, James; Löwe, Jan

    2018-04-01

    Most, if not all, bacterial and archaeal cells contain at least one protein filament system. Although these filament systems in some cases form structures that are very similar to eukaryotic cytoskeletons, the term 'prokaryotic cytoskeletons' is used to refer to many different kinds of protein filaments. Cytoskeletons achieve their functions through polymerization of protein monomers and the resulting ability to access length scales larger than the size of the monomer. Prokaryotic cytoskeletons are involved in many fundamental aspects of prokaryotic cell biology and have important roles in cell shape determination, cell division and nonchromosomal DNA segregation. Some of the filament-forming proteins have been classified into a small number of conserved protein families, for example, the almost ubiquitous tubulin and actin superfamilies. To understand what makes filaments special and how the cytoskeletons they form enable cells to perform essential functions, the structure and function of cytoskeletal molecules and their filaments have been investigated in diverse bacteria and archaea. In this Review, we bring these data together to highlight the diverse ways that linear protein polymers can be used to organize other molecules and structures in bacteria and archaea.

  16. Developmental pattern of the neuronal intermediate filament inaa in the zebrafish retina.

    Science.gov (United States)

    Liao, Meng-Lin; Peng, Wei-Hau; Kan, Daphne; Chien, Chung-Liang

    2016-12-15

    α-Internexin is a member of the neuronal intermediate filament (nIF) protein family, which also includes peripherin and neurofilament (NF) triplet proteins. Previous studies found that expression of α-internexin precedes that of the NF triplet proteins in mammals and suggested that α-internexin plays a key role in the neuronal cytoskeleton network during development. In this study, we aimed to analyze the expression patterns and function of internexin neuronal intermediate filament protein-alpha a (inaa), the encoding gene of which is a homolog of the mammalian α-internexin, during retinal development in zebrafish. Via in vitro and in vivo studies, we demonstrated that zebrafish inaa is an α-internexin homolog that shares characteristics with nIFs. An immunohistochemical analysis of zebrafish revealed that inaa was distributed dynamically in the developing retina. It was widely localized in retinal neuroepithelial cells at 1 day postfertilization (dpf), and was mainly found in the ganglion cell layer (GCL) and inner part of the inner nuclear layer (INL) from 3-9 dpf; after 14 dpf, it was restricted to the outer nuclear layer (ONL). Moreover, we demonstrated for the first time that inaa acted distinctively from the cytoskeletal scaffold of zebrafish cone photoreceptors during development. In conclusion, we demonstrated the morphological features of a novel nIF, inaa, and illustrated its developmental expression pattern in the zebrafish retina. J. Comp. Neurol. 524:3810-3826, 2016. © 2016 Wiley Periodicals, Inc. © 2016 Wiley Periodicals, Inc.

  17. In Vitro Assembly Kinetics of Cytoplasmic Intermediate Filaments: A Correlative Monte Carlo Simulation Study.

    Directory of Open Access Journals (Sweden)

    Norbert Mücke

    Full Text Available Intermediate filament (IF elongation proceeds via full-width "mini-filaments", referred to as "unit-length" filaments (ULFs, which instantaneously form by lateral association of extended coiled-coil complexes after assembly is initiated. In a comparatively much slower process, ULFs longitudinally interact end-to-end with other ULFs to form short filaments, which further anneal with ULFs and with each other to increasingly longer filaments. This assembly concept was derived from time-lapse electron and atomic force microscopy data. We previously have quantitatively verified this concept through the generation of time-dependent filament length-profiles and an analytical model that describes assembly kinetics well for about the first ten minutes. In this time frame, filaments are shorter than one persistence length, i.e. ~1 μm, and thus filaments were treated as stiff rods associating via their ends. However, when filaments grow several μm in length over hours, their flexibility becomes a significant factor for the kinetics of the longitudinal annealing process. Incorporating now additional filament length distributions that we have recorded after extended assembly times by total internal reflection fluorescence microscopy (TIRFM, we developed a Monte Carlo simulation procedure that accurately describes the underlying assembly kinetics for large time scales.

  18. Engineering of filamentous bacteriophage for protein sensing

    Science.gov (United States)

    Brasino, Michael

    Methods of high throughput, sensitive and cost effective quantification of proteins enables personalized medicine by allowing healthcare professionals to better monitor patient condition and response to treatment. My doctoral research has attempted to advance these methods through the use of filamentous bacteriophage (phage). These bacterial viruses are particularly amenable to both genetic and chemical engineering and can be produced efficiently in large amounts. Here, I discuss several strategies for modifying phage for use in protein sensing assays. These include the expression of bio-orthogonal conjugation handles on the phage coat, the incorporation of specific recognition sequences within the phage genome, and the creation of antibody-phage conjugates via a photo-crosslinking non-canonical amino acid. The physical and chemical characterization of these engineered phage and the results of their use in modified protein sensing assays will be presented.

  19. Pathomorphological feature of chronic pancreatitis (CP is the development of pancreatic fibrosis with the accumulation of various collagen types, tubulin, fibronectin, laminin, and also intermediate filament proteins produced by activated pancreatic stel

    Directory of Open Access Journals (Sweden)

    T. V. Turovskaya

    2013-04-01

    Full Text Available T. V. Turovskaya, A. M. Gnilorybov, L. V. Vasilyeva Pathomorphological feature of chronic pancreatitis (CP is the development of pancreatic fibrosis with the accumulation of various collagen types, tubulin, fibronectin, laminin, and also intermediate filament proteins produced by activated pancreatic stellate cells (PSCs, which express the cytoskeletal α-smooth muscle actin (α-SMA. The aim of the research: determination of immunophenotype and proliferative activity of pancreatic stellate cells as well as the main histotopographic components of severe pancreatic fibrosis and accumulation of collagen I, III and IV types in pancreas at CP. Materials and methods. Histological, histochemical (Van Gieson's and Masson's trichrome staining, immunohistochemical (α-SMA, vimentin, desmin, fibronectin, Ki-67, collagen I, III and IV types and morphometric studies (Image J program of accumulation of various collagen types, represented in standard unit of optical density (SUOD, were held at pancreas biopsies of 30 patients (35-72 years old with CP. Results. It was found that development of severe pancreatic fibrosis is promoted by proliferation and increase of α-SMA+, vimentin+, desmin+ activated stellate cells and deposition of significant amount of collagen I, III, IV types and fibronectin in pancreas that are synthesized by PSCs. In areas of severe fibrosis Ki-67 expression is detected in the nuclei of at least 25% of PSCs, that corresponds to relatively low levels of proliferation. Four components of severe pancreatic fibrosis: circular-periductal fibrosis involving the large ducts of the pancreas, laminar fibrosis in extensive fibrous fields between large ducts and acinar tissue, as well as tape-like interlobular and septal-periacinar intralobular pancreatic fibrosis are identified in patients with CP. Conclusion. Morphological manifestation of severe circular-periductal pancreatic fibrosis is the presence of significant concentric fibrosis around the

  20. Physical principles of filamentous protein self-assembly kinetics

    International Nuclear Information System (INIS)

    Michaels, Thomas C T; Liu, Lucie X; Meisl, Georg; Knowles, Tuomas P J

    2017-01-01

    The polymerization of proteins and peptides into filamentous supramolecular structures is an elementary form of self-organization of key importance to the functioning biological systems, as in the case of actin biofilaments that compose the cellular cytoskeleton. Aberrant filamentous protein self-assembly, however, is associated with undesired effects and severe clinical disorders, such as Alzheimer’s and Parkinson’s diseases, which, at the molecular level, are associated with the formation of certain forms of filamentous protein aggregates known as amyloids. Moreover, due to their unique physicochemical properties, protein filaments are finding extensive applications as biomaterials for nanotechnology. With all these different factors at play, the field of filamentous protein self-assembly has experienced tremendous activity in recent years. A key question in this area has been to elucidate the microscopic mechanisms through which filamentous aggregates emerge from dispersed proteins with the goal of uncovering the underlying physical principles. With the latest developments in the mathematical modeling of protein aggregation kinetics as well as the improvement of the available experimental techniques it is now possible to tackle many of these complex systems and carry out detailed analyses of the underlying microscopic steps involved in protein filament formation. In this paper, we review some classical and modern kinetic theories of protein filament formation, highlighting their use as a general strategy for quantifying the molecular-level mechanisms and transition states involved in these processes. (topical review)

  1. A network of 2-4 nm filaments found in sea urchin smooth muscle. Protein constituents and in situ localization.

    Science.gov (United States)

    Pureur, R P; Coffe, G; Soyer-Gobillard, M O; de Billy, F; Pudles, J

    1986-01-01

    In this report the coisolation of two proteins from sea urchin smooth muscle of apparent molecular weights (Mr) 54 and 56 kD respectively, as determined on SDS-PAGE, is described. Like the intermediate filament proteins, these two proteins are insoluble in high ionic strength buffer solution. On two-dimensional gel electrophoresis and by immunological methods it is shown that these proteins are not related (by these criteria) to rat smooth muscle desmin (54 kD) or vimentin (56 kD). Furthermore, in conditions where both desmin and vimentin assemble in vitro into 10 nm filaments, the sea urchin smooth muscle proteins do not assemble into filaments. Ultrastructural studies on the sea urchin smooth muscle cell show that the thin and thick filaments organization resembles that described in the vertebrate smooth muscle. However, instead of 10 nm filaments, a network of filaments, 2-4 nm in diameter, is revealed, upon removal of the thin and thick filaments by 0.6 M KCl treatment. By indirect immunofluorescence microscopy, and in particular by immunocytochemical electron microscopy studies on the sea urchin smooth muscle cell, it is shown that the antibodies raised against both 54 and 56 kD proteins appear to specifically label these 2-4 nm filaments. These findings indicate that both the 54 and 56 kD proteins might be constituents of this category of filaments. The possible significance of this new cytoskeletal element, that we have named echinonematin filaments, is discussed.

  2. Neuronal intermediate filaments in the developing tongue of the frog Rana esculenta

    Directory of Open Access Journals (Sweden)

    K Zuwala

    2009-06-01

    Full Text Available The expression of several neuronal intermediate filament (NIF proteins was investigated in the tongue of metamorphosing tadpoles (stage 38-45 of Gosner and in adult individuals of the frog, Rana esculenta by means of immunohistochemistry. Results showed that nerve fibres at early stages of tongue development expressed peripherin (a NIF protein usually found in differentiating neurones as well as the light- and medium molecular weight NIF polypeptide subunits (NF-L and NF-M, respectively; in the adult frog, peripherin was still found in nerve fibres reaching the fungiform papilla together with NF-M, but NF-L immunoreactivity was absent therein. Clusters of epithelial cells expressing peripherin were found in the early developing tongue before differentiation of taste organs, and NF-L and NF-H immunoreactivities were present in basal (Merkel cells of the adult frog taste disc. Results indicate that neurones innervating the adult frog’s taste disc maintain a certain plasticity in their cytoskeleton and that neuronal-like cells are present in the undifferentiated and differentiated tongue epithelium possibly playing a role in the developing and mature taste organ.

  3. Intermediate filament immunohistochemistry of astroglial cells in the leopard gecko, Eublepharis macularius.

    Science.gov (United States)

    Lazzari, Maurizio; Franceschini, Valeria

    2005-11-01

    The distribution of intermediate filament molecular markers, glial fibrillary acidic protein (GFAP) and vimentin, has been studied in the central nervous system (CNS) of the adult leopard gecko, Eublepharis macularius. This immunohistochemical study points out the presence of different astroglial cell types. The main pattern is constituted by ependymal radial glia, which have their cell bodies located in the ependymal layer throughout the brain ventricular system. Radial glia proper or radial astrocytes show their cell bodies displaced from the ependymal layer into a periependymal zone and are observed only in the spinal cord. Star-shaped astrocytes are scarce. They are detected in the ventral and lateral regions of the diencephalon and mesencephalon, in the superficial layer of the optic tectum, in the ventral medulla oblongata, and in the ventral and lateral spinal cord. In the different regions of the CNS, the staining intensity appears not to be identical even in the same cellular type. The results reported in the present study show an heterogeneous feature of the astroglial pattern in E. macularius.

  4. Using Data Mining and Computational Approaches to Study Intermediate Filament Structure and Function.

    Science.gov (United States)

    Parry, David A D

    2016-01-01

    Experimental and theoretical research aimed at determining the structure and function of the family of intermediate filament proteins has made significant advances over the past 20 years. Much of this has either contributed to or relied on the amino acid sequence databases that are now available online, and the data mining approaches that have been developed to analyze these sequences. As the quality of sequence data is generally high, it follows that it is the design of the computational and graphical methodologies that are of especial importance to researchers who aspire to gain a greater understanding of those sequence features that specify both function and structural hierarchy. However, these techniques are necessarily subject to limitations and it is important that these be recognized. In addition, no single method is likely to be successful in solving a particular problem, and a coordinated approach using a suite of methods is generally required. A final step in the process involves the interpretation of the results obtained and the construction of a working model or hypothesis that suggests further experimentation. While such methods allow meaningful progress to be made it is still important that the data are interpreted correctly and conservatively. New data mining methods are continually being developed, and it can be expected that even greater understanding of the relationship between structure and function will be gleaned from sequence data in the coming years. Copyright © 2016 Elsevier Inc. All rights reserved.

  5. Vimentin intermediate filaments template microtubule networks to enhance persistence in cell polarity and directed migration

    OpenAIRE

    Gan, Zhuo; Ding, Liya; Burckhardt, Christoph J.; Lowery, Jason; Zaritsky, Assaf; Sitterley, Karlyndsay; Mota, Andressa; Costigliola, Nancy; Starker, Colby G.; Voytas, Daniel F.; Tytell, Jessica; Goldman, Robert D.; Danuser, Gaudenz

    2016-01-01

    Increased expression of vimentin intermediate filaments (VIF) enhances directed cell migration, but the mechanism behind VIF’s effect on motility is not understood. VIF interact with microtubules, whose organization contributes to polarity maintenance in migrating cells. Here we characterize the dynamic coordination of VIF and microtubule networks in wounded monolayers of Retinal Pigment Epithelial cells. By genome editing we fluorescently labelled endogenous vimentin and α-...

  6. α-Internexin aggregates are abundant in neuronal intermediate filament inclusion disease (NIFID) but rare in other neurodegenerative diseases

    Science.gov (United States)

    Cairns, Nigel J.; Uryu, Kunihiro; Bigio, Eileen H.; Mackenzie, Ian R. A.; Gearing, Marla; Duyckaerts, Charles; Yokoo, Hideaki; Nakazato, Yoichi; Jaros, Evelyn; Perry, Robert H.; Arnold, Steven E.; Lee, Virginia M.-Y.; Trojanowski, John Q.

    2009-01-01

    Abnormal neuronal aggregates of α-internexin and the three neurofilament (NF) subunits, NF-L, NF-M, and NF-H have recently been identified as the pathological hallmarks of neuronal intermediate filament (IF) inclusion disease (NIFID), a novel neurological disease of early onset with a variable clinical phenotype including frontotemporal dementia, pyramidal and extrapyramidal signs. α-Internexin, a class IV IF protein, a major component of inclusions in NIFID, has not previously been identified as a component of the pathological protein aggregates of any other neurodegenerative disease. Therefore, to determine the specificity of this protein, α-internexin immunohistochemistry was undertaken on cases of NIFID, non-tau frontotemporal dementias, motor neuron disease, α-synucleinopathies, tauopathies, and normal aged control brains. Our results indicate that class IV IF proteins are present within the pleomorphic inclusions of all cases of NIFID. Small subsets of abnormal neuronal inclusions in Alzheimer's disease, Lewy body diseases, and motor neuron disease also contain epitopes of α-internexin. Thus, α-internexin is a major component of the neuronal inclusions in NIFID and a relatively minor component of inclusions in other neurodegenerative diseases. The discovery of α-internexin in neuronal cytoplasmic inclusions implicates novel mechanisms of pathogenesis in NIFID and other neurological diseases with pathological filamentous neuronal inclusions. PMID:15170578

  7. Controlling skeletal muscle CPT-I malonyl-CoA sensitivity: the importance of AMPK-independent regulation of intermediate filaments during exercise.

    Science.gov (United States)

    Miotto, Paula M; Steinberg, Gregory R; Holloway, Graham P

    2017-02-15

    The obligatory role of carnitine palmitoyltransferase-I (CPT-I) in mediating mitochondrial lipid transport is well established, a process attenuated by malonyl-CoA (M-CoA). However, the necessity of reducing M-CoA concentrations to promote lipid oxidation has recently been challenged, suggesting external regulation on CPT-I. Since previous work in hepatocytes suggests the involvement of the intermediate filament fraction of the cytoskeleton in regulating CPT-I, we investigated in skeletal muscle if CPT-I sensitivity for M-CoA inhibition could be regulated by the intermediate filaments, and whether AMP-activated protein kinase (AMPK) could be involved in this process. Chemical disruption (3,3'-iminodipropionitrile, IDPN) of the intermediate filaments did not alter mitochondrial respiration or sensitivity for numerous substrates (palmitoyl-CoA, ADP, palmitoyl carnitine and pyruvate). In contrast, IDPN reduced CPT-I sensitivity for M-CoA inhibition in permeabilized muscle fibers, identifying M-CoA kinetics as a specific target for intermediate filament regulation. Importantly, exercise mimicked the effect of IDPN on M-CoA sensitivity, suggesting that intermediate filament disruption in vivo is physiologically important for CPT-I regulation. To ascertain a potential mechanism, since AMPK is activated during exercise, AMPK β1β2-KO mice were utilized in an attempt to ablate the observed exercise response. Unexpectedly, these mice displayed drastic attenuation in resting M-CoA sensitivity, such that exercise and IDPN could not further alter M-CoA sensitivity. These data suggest that AMPK is not required for the regulation of the intermediate filament interaction with CPT-I. Altogether, these data highlight that M-CoA sensitivity is important for regulating mitochondrial lipid transport. Moreover, M-CoA sensitivity appears to be regulated by intermediate filament interaction with CPT-I, a process that is important when metabolic homeostasis is challenged. © 2017 The

  8. The dermatan sulfate proteoglycan decorin modulates α2β1 integrin and the vimentin intermediate filament system during collagen synthesis.

    Directory of Open Access Journals (Sweden)

    Oliver Jungmann

    Full Text Available Decorin, a small leucine-rich proteoglycan harboring a dermatan sulfate chain at its N-terminus, is involved in regulating matrix organization and cell signaling. Loss of the dermatan sulfate of decorin leads to an Ehlers-Danlos syndrome characterized by delayed wound healing. Decorin-null (Dcn(-/- mice display a phenotype similar to that of EDS patients. The fibrillar collagen phenotype of Dcn(-/- mice could be rescued in vitro by decorin but not with decorin lacking the glycosaminoglycan chain. We utilized a 3D cell culture model to investigate the impact of the altered extracellular matrix on Dcn(-/- fibroblasts. Using 2D gel electrophoresis followed by mass spectrometry, we identified vimentin as one of the proteins that was differentially upregulated by the presence of decorin. We discovered that a decorin-deficient matrix leads to abnormal nuclear morphology in the Dcn(-/- fibroblasts. This phenotype could be rescued by the decorin proteoglycan but less efficiently by the decorin protein core. Decorin treatment led to a significant reduction of the α2β1 integrin at day 6 in Dcn(-/- fibroblasts, whereas the protein core had no effect on β1. Interestingly, only the decorin core induced mRNA synthesis, phosphorylation and de novo synthesis of vimentin indicating that the proteoglycan decorin in the extracellular matrix stabilizes the vimentin intermediate filament system. We could support these results in vivo, because the dermis of wild-type mice have more vimentin and less β1 integrin compared to Dcn(-/-. Furthermore, the α2β1 null fibroblasts also showed a reduced amount of vimentin compared to wild-type. These data show for the first time that decorin has an impact on the biology of α2β1 integrin and the vimentin intermediate filament system. Moreover, our findings provide a mechanistic explanation for the reported defects in wound healing associated with the Dcn(-/- phenotype.

  9. Airborne acrolein induces keratin-8 (Ser-73) hyperphosphorylation and intermediate filament ubiquitination in bronchiolar lung cell monolayers.

    Science.gov (United States)

    Burcham, Philip C; Raso, Albert; Henry, Peter J

    2014-05-07

    The combustion product acrolein is a key mediator of pulmonary edema in victims of smoke inhalation injury. Since studying acrolein toxicity in conventional in vitro systems is complicated by reactivity with nucleophilic culture media constituents, we explored an exposure system which delivers airborne acrolein directly to lung cell monolayers at the air-liquid interface. Calu-3 lung adenocarcinoma cells were maintained on membrane inserts such that the basal surface was bathed in nucleophile-free media while the upper surface remained in contact with acrolein-containing air. Cells were exposed to airborne acrolein for 30 min before they were allowed to recover in fresh media, with cell sampling at defined time points to allow evaluation of toxicity and protein damage. After prior exposure to acrolein, cell ATP levels remained close to controls for 4h but decreased in an exposure-dependent manner by 24h. A loss of transepithelial electrical resistance and increased permeability to fluorescein isothiocyanate-labeled dextran preceded ATP loss. Use of antibody arrays to monitor protein expression in exposed monolayers identified strong upregulation of phospho-keratin-8 (Ser(73)) as an early consequence of acrolein exposure. These changes were accompanied by chemical damage to keratin-8 and other intermediate filament family members, while acrolein exposure also resulted in controlled ubiquitination of high mass proteins within the intermediate filament extracts. These findings confirm the usefulness of systems allowing delivery of airborne smoke constituents to lung cell monolayers during studies of the molecular basis for acute smoke intoxication injury. Copyright © 2014 Elsevier Ireland Ltd. All rights reserved.

  10. Rice intermediate filament, OsIF, stabilizes photosynthetic machinery and yield under salinity and heat stress.

    Science.gov (United States)

    Soda, Neelam; Gupta, Brijesh K; Anwar, Khalid; Sharan, Ashutosh; Govindjee; Singla-Pareek, Sneh L; Pareek, Ashwani

    2018-03-06

    Cytoskeleton plays a vital role in stress tolerance; however, involvement of intermediate filaments (IFs) in such a response remains elusive in crop plants. This study provides clear evidence about the unique involvement of IFs in cellular protection against abiotic stress in rice. Transcript abundance of Oryza sativa intermediate filament (OsIF) encoding gene showed 2-10 fold up-regulation under different abiotic stress. Overexpression of OsIF in transgenic rice enhanced tolerance to salinity and heat stress, while its knock-down (KD) rendered plants more sensitive thereby indicating the role of IFs in promoting survival under stress. Seeds of OsIF overexpression rice germinated normally in the presence of high salt, showed better growth, maintained chloroplast ultrastructure and favourable K + /Na + ratio than the wild type (WT) and KD plants. Analysis of photosynthesis and chlorophyll a fluorescence data suggested better performance of both photosystem I and II in the OsIF overexpression rice under salinity stress as compared to the WT and KD. Under salinity and high temperature stress, OsIF overexpressing plants could maintain significantly high yield, while the WT and KD plants could not. Further, metabolite profiling revealed a 2-4 fold higher accumulation of proline and trehalose in OsIF overexpressing rice than WT, under salinity stress.

  11. The roles and regulation of the actin cytoskeleton, intermediate filaments and microtubules in smooth muscle cell migration.

    Science.gov (United States)

    Tang, Dale D; Gerlach, Brennan D

    2017-04-08

    Smooth muscle cell migration has been implicated in the development of respiratory and cardiovascular systems; and airway/vascular remodeling. Cell migration is a polarized cellular process involving a protrusive cell front and a retracting trailing rear. There are three cytoskeletal systems in mammalian cells: the actin cytoskeleton, the intermediate filament network, and microtubules; all of which regulate all or part of the migrated process. The dynamic actin cytoskeleton spatially and temporally regulates protrusion, adhesions, contraction, and retraction from the cell front to the rear. c-Abl tyrosine kinase plays a critical role in regulating actin dynamics and migration of airway smooth muscle cells and nonmuscle cells. Recent studies suggest that intermediate filaments undergo reorganization during migration, which coordinates focal adhesion dynamics, cell contraction, and nucleus rigidity. In particular, vimentin intermediate filaments undergo phosphorylation and reorientation in smooth muscle cells, which may regulate cell contraction and focal adhesion assembly/disassembly. Motile cells are characterized by a front-rear polarization of the microtubule framework, which regulates all essential processes leading to cell migration through its role in cell mechanics, intracellular trafficking, and signaling. This review recapitulates our current knowledge how the three cytoskeletal systems spatially and temporally modulate the migratory properties of cells. We also summarize the potential role of migration-associated biomolecules in lung and vascular diseases.

  12. Intermediate filaments of zebrafish retinal and optic nerve astrocytes and Müller glia: differential distribution of cytokeratin and GFAP

    Directory of Open Access Journals (Sweden)

    Mosier Amanda L

    2010-03-01

    Full Text Available Abstract Background Optic nerve regeneration (ONR following injury is a model for central nervous system regeneration. In zebrafish, ONR is rapid - neurites cross the lesion and enter the optic tectum within 7 days; in mammals regeneration does not take place unless astrocytic reactivity is suppressed. Glial fibrillary acidic protein (GFAP is used as a marker for retinal and optic nerve astrocytes in both fish and mammals, even though it has long been known that astrocytes of optic nerves in many fish, including zebrafish, express cytokeratins and not GFAP. We used immunofluorescence to localize GFAP and cytokeratin in wild-type zebrafish and transgenic zebrafish expressing green fluorescent protein (GFP under control of a GFAP promoter to determine the pattern of expression of intermediate filaments in retina and optic nerve. Findings GFAP labeling and GFAP gene expression as indicated by GFP fluorescence was found only in the Müller glial cells of the retina. Within Müller cells, GFP fluorescence filled the entire cell while GFAP labelling was more restricted in distribution. No GFAP expression was observed in optic nerves. Cytokeratin labeling of astrocytes was observed throughout the optic nerve and less intensely in cells in the retinal inner plexiform layer. The retinal inner limiting membrane was strongly labeled by anti-cytokeratin. Conclusions Studies of astrocyte function during ONR in zebrafish cannot solely rely on GFAP as an astrocyte marker or indicator of reactivity. Future studies of ONR in zebrafish should include evaluation of changes in cytokeratin expression and localization in the optic nerve.

  13. Mechanosensitive kinetic preference of actin-binding protein to actin filament.

    Science.gov (United States)

    Inoue, Yasuhiro; Adachi, Taiji

    2016-04-01

    The kinetic preference of actin-binding proteins to actin filaments is altered by external forces on the filament. Such an altered kinetic preference is largely responsible for remodeling the actin cytoskeletal structure in response to intracellular forces. During remodeling, actin-binding proteins and actin filaments interact under isothermal conditions, because the cells are homeostatic. In such a temperature homeostatic state, we can rigorously and thermodynamically link the chemical potential of actin-binding proteins to stresses on the actin filaments. From this relationship, we can construct a physical model that explains the force-dependent kinetic preference of actin-binding proteins to actin filaments. To confirm the model, we have analyzed the mechanosensitive alternation of the kinetic preference of Arp2/3 and cofilin to actin filaments. We show that this model captures the qualitative responses of these actin-binding proteins to the forces, as observed experimentally. Moreover, our theoretical results demonstrate that, depending on the structural parameters of the binding region, actin-binding proteins can show different kinetic responses even to the same mechanical signal tension, in which the double-helix nature of the actin filament also plays a critical role in a stretch-twist coupling of the filament.

  14. Analysis of the Yeast Kinome Reveals a Network of Regulated Protein Localization during Filamentous Growth

    OpenAIRE

    Bharucha, Nikë; Ma, Jun; Dobry, Craig J.; Lawson, Sarah K.; Yang, Zhifen; Kumar, Anuj

    2008-01-01

    The subcellular distribution of kinases and other signaling proteins is regulated in response to cellular cues; however, the extent of this regulation has not been investigated for any gene set in any organism. Here, we present a systematic analysis of protein kinases in the budding yeast, screening for differential localization during filamentous growth. Filamentous growth is an important stress response involving mitogen-activated protein kinase and cAMP-dependent protein kinase signaling m...

  15. Chaperonin filaments: The archael cytoskeleton

    Energy Technology Data Exchange (ETDEWEB)

    Trent, J.D.; Kagawa, H.K.; Yaoi, Takuro; Olle, E.; Zaluzec, N.J.

    1997-08-01

    Chaperonins are multi-subunit double-ring complexed composed of 60-kDa proteins that are believed to mediate protein folding in vivo. The chaperonins in the hyperthermophilic archaeon Sulfolobus shibatae are composed of the organism`s two most abundant proteins, which represent 4% of its total protein and have an intracellular concentration of {ge} 3.0 mg/ml. At concentrations of 1.0 mg/ml, purified chaperonin proteins aggregate to form ordered filaments. Filament formation, which requires Mg{sup ++} and nucleotide binding (not hydrolysis), occurs at physiological temperatures under conditions suggesting filaments may exist in vivo. If the estimated 4,600 chaperonins per cell, formed filaments in vivo, they could create a matrix of filaments that would span the diameter of an average S. shibatae cell 100 times. Direct observations of unfixed, minimally treated cells by intermediate voltage electron microscopy (300 kV) revealed an intracellular network of filaments that resembles chaperonin filaments produced in vitro. The hypothesis that the intracellular network contains chaperonins is supported by immunogold analyses. The authors propose that chaperonin activity may be regulated in vivo by filament formation and that chaperonin filaments may serve a cytoskeleton-like function in archaea and perhaps in other prokaryotes.

  16. RecA-DNA filament topology: the overlooked alternative of an unconventional syn-syn duplex intermediate

    DEFF Research Database (Denmark)

    Egel, Richard

    2007-01-01

    The helical filaments of RecA protein mediate strand exchange for homologous recombination, but the paths of the interacting DNAs have yet to be determined. Although this interaction is commonly limited to three strands, it is reasoned here that the intrinsic symmetry relationships of quadruplex...... topology are superior in explaining a range of observations. In particular, this topology suggests the potential of post-exchange base pairing in the unorthodox configuration of syn-syn glycosidic bonds between the nucleotide bases and the pentose rings in the sugar-phosphate backbone, which would...

  17. Metal ion-dependent, reversible, protein filament formation by designed beta-roll polypeptides

    Science.gov (United States)

    Scotter, Andrew J; Guo, Meng; Tomczak, Melanie M; Daley, Margaret E; Campbell, Robert L; Oko, Richard J; Bateman, David A; Chakrabartty, Avijit; Sykes, Brian D; Davies, Peter L

    2007-01-01

    Background A right-handed, calcium-dependent β-roll structure found in secreted proteases and repeat-in-toxin proteins was used as a template for the design of minimal, soluble, monomeric polypeptides that would fold in the presence of Ca2+. Two polypeptides were synthesised to contain two and four metal-binding sites, respectively, and exploit stacked tryptophan pairs to stabilise the fold and report on the conformational state of the polypeptide. Results Initial analysis of the two polypeptides in the presence of calcium suggested the polypeptides were disordered. The addition of lanthanum to these peptides caused aggregation. Upon further study by right angle light scattering and electron microscopy, the aggregates were identified as ordered protein filaments that required lanthanum to polymerize. These filaments could be disassembled by the addition of a chelating agent. A simple head-to-tail model is proposed for filament formation that explains the metal ion-dependency. The model is supported by the capping of one of the polypeptides with biotin, which disrupts filament formation and provides the ability to control the average length of the filaments. Conclusion Metal ion-dependent, reversible protein filament formation is demonstrated for two designed polypeptides. The polypeptides form filaments that are approximately 3 nm in diameter and several hundred nm in length. They are not amyloid-like in nature as demonstrated by their behaviour in the presence of congo red and thioflavin T. A capping strategy allows for the control of filament length and for potential applications including the "decoration" of a protein filament with various functional moieties. PMID:17908326

  18. Filamentous fungi as cell factories for heterologous protein production

    NARCIS (Netherlands)

    Punt, P.J.; Biezen, N. van; Conesa, A.; Albers, A.; Mangnus, J.; Hondel, C. van den

    2002-01-01

    Filamentous fungi have been used as sources of metabolites and enzymes for centuries. For about two decades, molecular genetic tools have enabled us to use these organisms to express extra copies of both endogenous and exogenous genes. This review of current practice reveals that molecular tools

  19. Protein secretion in the filamentous fungus Aspergillus niger

    NARCIS (Netherlands)

    Weenink, Xavier Oswin

    2008-01-01

    Filamentous fungi are multicellular eukaryotic organisms, which represent a separate taxonomic group organisms within the fungal kingdom, apart from the yeasts. These fungi always need a substrate to grow on, this can be living or dead material. Fungi possess the capacity to secrete high levels of

  20. Filamentous sieve element proteins are able to limit phloem mass flow, but not phytoplasma spread.

    Science.gov (United States)

    Pagliari, Laura; Buoso, Sara; Santi, Simonetta; Furch, Alexandra C U; Martini, Marta; Degola, Francesca; Loschi, Alberto; van Bel, Aart J E; Musetti, Rita

    2017-06-15

    In Fabaceae, dispersion of forisomes-highly ordered aggregates of sieve element proteins-in response to phytoplasma infection was proposed to limit phloem mass flow and, hence, prevent pathogen spread. In this study, the involvement of filamentous sieve element proteins in the containment of phytoplasmas was investigated in non-Fabaceae plants. Healthy and infected Arabidopsis plants lacking one or two genes related to sieve element filament formation-AtSEOR1 (At3g01680), AtSEOR2 (At3g01670), and AtPP2-A1 (At4g19840)-were analysed. TEM images revealed that phytoplasma infection induces phloem protein filament formation in both the wild-type and mutant lines. This result suggests that, in contrast to previous hypotheses, sieve element filaments can be produced independently of AtSEOR1 and AtSEOR2 genes. Filament presence was accompanied by a compensatory overexpression of sieve element protein genes in infected mutant lines in comparison with wild-type lines. No correlation was found between phloem mass flow limitation and phytoplasma titre, which suggests that sieve element proteins are involved in defence mechanisms other than mechanical limitation of the pathogen. © The Author 2017. Published by Oxford University Press on behalf of the Society for Experimental Biology.

  1. Medium Calcium Concentration Determines Keratin Intermediate Filament Density and Distribution in Immortalized Cultured Thymic Epithelial Cells (TECs)

    Science.gov (United States)

    Sands, Sandra S.; Meek, William D.; Hayashi, Jun; Ketchum, Robert J.

    2005-08-01

    Isolation and culture of thymic epithelial cells (TECs) using conventional primary tissue culture techniques under conditions employing supplemented low calcium medium yielded an immortalized cell line derived from the LDA rat (Lewis [Rt1l] cross DA [Rt1a]) that could be manipulated in vitro. Thymi were harvested from 4 5-day-old neonates, enzymically digested using collagenase (1 mg/ml, 37°C, 1 h) and cultured in low calcium WAJC404A medium containing cholera toxin (20 ng/ml), dexamethasone (10 nM), epidermal growth factor (10 ng/ml), insulin (10 [mu]g/ml), transferrin (10 [mu]g/ml), 2% calf serum, 2.5% Dulbecco's Modified Eagle's Medium (DMEM), and 1% antibiotic/antimycotic. TECs cultured in low calcium displayed round to spindle-shaped morphology, distinct intercellular spaces (even at confluence), and dense reticular-like keratin patterns. In high calcium (0.188 mM), TECs formed cobblestone-like confluent monolayers that were resistant to trypsinization (0.05%) and displayed keratin intermediate filaments concentrated at desmosomal junctions between contiguous cells. Changes in cultured TEC morphology were quantified by an analysis of desmosome/membrane relationships in high and low calcium media. Desmosomes were significantly increased in the high calcium medium. These studies may have value when considering the growth conditions of cultured primary cell lines like TECs.

  2. Superhelical architecture of the myosin filament-linking protein myomesin with unusual elastic properties.

    Directory of Open Access Journals (Sweden)

    Nikos Pinotsis

    2012-02-01

    Full Text Available Active muscles generate substantial mechanical forces by the contraction/relaxation cycle, and, to maintain an ordered state, they require molecular structures of extraordinary stability. These forces are sensed and buffered by unusually long and elastic filament proteins with highly repetitive domain arrays. Members of the myomesin protein family function as molecular bridges that connect major filament systems in the central M-band of muscle sarcomeres, which is a central locus of passive stress sensing. To unravel the mechanism of molecular elasticity in such filament-connecting proteins, we have determined the overall architecture of the complete C-terminal immunoglobulin domain array of myomesin by X-ray crystallography, electron microscopy, solution X-ray scattering, and atomic force microscopy. Our data reveal a dimeric tail-to-tail filament structure of about 360 Å in length, which is folded into an irregular superhelical coil arrangement of almost identical α-helix/domain modules. The myomesin filament can be stretched to about 2.5-fold its original length by reversible unfolding of these linkers, a mechanism that to our knowledge has not been observed previously. Our data explain how myomesin could act as a highly elastic ribbon to maintain the overall structural organization of the sarcomeric M-band. In general terms, our data demonstrate how repetitive domain modules such as those found in myomesin could generate highly elastic protein structures in highly organized cell systems such as muscle sarcomeres.

  3. Effect of desynchronosis on oxidative stress biomarkers and the state of glial intermediate filaments in the brains of rats subject to aging

    Directory of Open Access Journals (Sweden)

    S. Kyrychenko

    2016-09-01

    Full Text Available Desynchronosis may be the cause of many diseases. Oxidative stress plays an important role in the pathogenesis of various diseases. The present study investigates the effect of constant light on biomarkers of oxidative stress and content of glial intermediate filaments protein in the brains of old rats. We found that desynchronosis led to development of oxidative stress in the hippocampus, cerebral cortex and cerebellum of old rats. Prolonged continuous lighting led to an increase in the content of TBA-reactive products in all studied regions of the brains of old rats. This indicates an activation of lipid peroxidation and oxidative stress. Significant changes in the content of TBA-reactive products were found in the departments responsible for the functions of the higher nervous activity, particularly in the hippocampus and cerebral cortex. The level of restored glutathione in all three regions of the brain decreased in the group of rats kept under constant illumination in comparison with the control group. The results of the indicators of locomotor and orienting-investigative activity of the animals in the "open field" test revealled changes in the indices for desynchronosis. This showed a reduction in locomotor activity, inhibition of exploratory activity and development of emotional stress. In the brains of old rats kept under constant illumination a significant increase in the content of glial fibrillary protein (GFAP was shown. There was a significant increase in the intensity of the polypeptide zone 49 kDa in the filamentous and soluble fraction of the cerebellum and hippocampus. This fact indicates that desynchronosis activates fibrillogenesis in glial cells. At the same time, there is degradation of polypeptides GFAP with Mr in the field of 46 kDa. Melatonin is a universal adaptogen that regulates the function of many body systems. The amount of melatonin which is synthesized depends on the illumination mode. Violation of the global

  4. Planck intermediate results XXXIII. Signature of the magnetic field geometry of interstellar filaments in dust polarization maps

    DEFF Research Database (Denmark)

    Ade, P. A. R.; Aghanim, N.; Alves, M. I. R.

    2016-01-01

    Planck observations at 353 GHz provide the first fully sampled maps of the polarized dust emission towards interstellar filaments and their backgrounds (i.e., the emission observed in the surroundings of the filaments). The data allow us to determine the intrinsic polarization properties of the f......Planck observations at 353 GHz provide the first fully sampled maps of the polarized dust emission towards interstellar filaments and their backgrounds (i.e., the emission observed in the surroundings of the filaments). The data allow us to determine the intrinsic polarization properties...

  5. Identification of intermediate species in protein-folding by ...

    Indian Academy of Sciences (India)

    -states, namely the native (N) and unfolded (U) forms of the protein present at any condition of the solvent, from a situation wherein intermediate state(s) could also be present. This differentiation of a two-state from a multi-state structural ...

  6. Cytoplasmic Dynein Is Required for the Spatial Organization of Protein Aggregates in Filamentous Fungi

    Directory of Open Access Journals (Sweden)

    Martin J. Egan

    2015-04-01

    Full Text Available Eukaryotes have evolved multiple strategies for maintaining cellular protein homeostasis. One such mechanism involves neutralization of deleterious protein aggregates via their defined spatial segregation. Here, using the molecular disaggregase Hsp104 as a marker for protein aggregation, we describe the spatial and temporal dynamics of protein aggregates in the filamentous fungus Aspergillus nidulans. Filamentous fungi, such as A. nidulans, are a diverse group of species of major health and economic importance and also serve as model systems for studying highly polarized eukaryotic cells. We find that microtubules promote the formation of Hsp104-positive aggregates, which coalesce into discrete subcellular structures in a process dependent on the microtubule-based motor cytoplasmic dynein. Finally, we find that impaired clearance of these inclusions negatively impacts retrograde trafficking of endosomes, a conventional dynein cargo, indicating that microtubule-based transport can be overwhelmed by chronic cellular stress.

  7. Human muscle LIM protein dimerizes along the actin cytoskeleton and cross-links actin filaments.

    Science.gov (United States)

    Hoffmann, Céline; Moreau, Flora; Moes, Michèle; Luthold, Carole; Dieterle, Monika; Goretti, Emeline; Neumann, Katrin; Steinmetz, André; Thomas, Clément

    2014-08-01

    The muscle LIM protein (MLP) is a nucleocytoplasmic shuttling protein playing important roles in the regulation of myocyte remodeling and adaptation to hypertrophic stimuli. Missense mutations in human MLP or its ablation in transgenic mice promotes cardiomyopathy and heart failure. The exact function(s) of MLP in the cytoplasmic compartment and the underlying molecular mechanisms remain largely unknown. Here, we provide evidence that MLP autonomously binds to, stabilizes, and bundles actin filaments (AFs) independently of calcium and pH. Using total internal reflection fluorescence microscopy, we have shown how MLP cross-links actin filaments into both unipolar and mixed-polarity bundles. Quantitative analysis of the actin cytoskeleton configuration confirmed that MLP substantially promotes actin bundling in live myoblasts. In addition, bimolecular fluorescence complementation (BiFC) assays revealed MLP self-association. Remarkably, BiFC complexes mostly localize along actin filament-rich structures, such as stress fibers and sarcomeres, supporting a functional link between MLP self-association and actin cross-linking. Finally, we have demonstrated that MLP self-associates through its N-terminal LIM domain, whereas it binds to AFs through its C-terminal LIM domain. Together our data support that MLP contributes to the maintenance of cardiomyocyte cytoarchitecture by a mechanism involving its self-association and actin filament cross-linking. Copyright © 2014, American Society for Microbiology. All Rights Reserved.

  8. Target Molecular Simulations of RecA Family Protein Filaments

    Directory of Open Access Journals (Sweden)

    Yeng-Tseng Wang

    2012-06-01

    Full Text Available Modeling of the RadA family mechanism is crucial to understanding the DNA SOS repair process. In a 2007 report, the archaeal RadA proteins function as rotary motors (linker region: I71-K88 such as shown in Figure 1. Molecular simulations approaches help to shed further light onto this phenomenon. We find 11 rotary residues (R72, T75-K81, M84, V86 and K87 and five zero rotary residues (I71, K74, E82, R83 and K88 in the simulations. Inclusion of our simulations may help to understand the RadA family mechanism.

  9. Estrous cycle-dependent changes of Fas expression in the bovine corpus luteum: influence of keratin 8/18 intermediate filaments and cytokines

    Directory of Open Access Journals (Sweden)

    Duncan Alice

    2012-10-01

    Full Text Available Abstract Background Fas expression and Fas-induced apoptosis are mechanisms attributed to the selective destruction of cells of the corpus luteum (CL during luteal regression. In certain cell-types, sensitivity to these death-inducing mechanisms is due to the loss or cleavage of keratin-containing intermediate filaments. Specifically, keratin 8/18 (K8/K18 filaments are hypothesized to influence cell death in part by regulating Fas expression at the cell surface. Methods Here, Fas expression on bovine luteal cells was quantified by flow cytometry during the early (Day 5, postovulation and late stages (Days 16–18, postovulation of CL function, and the relationship between Fas expression, K8/K18 filament expression and cytokine-induced cell death in vitro was evaluated. Results Both total and cell surface expression of Fas on luteal cells was greater for early versus late stage bovine CL (89% vs. 44% of cells for total Fas; 65% vs.18% of cells for cell surface Fas; respectively, P0.05, n=4 CL/stage, despite evidence these conditions increased Fas expression on HepG2 cells (P0.05 or stage of CL (P>0.05, n= 4 CL/stage on this outcome. Conclusion In conclusion, we rejected our null hypothesis that the cell surface expression of Fas does not differ between luteal cells of early and late stage CL. The results also did not support the idea that K8/K18 filaments influence the expression of Fas on the surface of bovine luteal cells. Potential downstream effects of these filaments on death signaling, however, remain a possibility. Importantly, the elevated expression of Fas observed on cells of early stage bovine CL compared to late stage bovine CL raises a provocative question concerning the physiological role(s of Fas in the corpus luteum, particularly during early luteal development.

  10. Molecular characterization of a phloem-specific gene encoding the filament protein, phloem protein 1 (PP1), from Cucurbita maxima.

    Science.gov (United States)

    Clark, A M; Jacobsen, K R; Bostwick, D E; Dannenhoffer, J M; Skaggs, M I; Thompson, G A

    1997-07-01

    Sieve elements in the phloem of most angiosperms contain proteinaceous filaments and aggregates called P-protein. In the genus Cucurbita, these filaments are composed of two major proteins: PP1, the phloem filament protein, and PP2, the phloem lactin. The gene encoding the phloem filament protein in pumpkin (Cucurbita maxima Duch.) has been isolated and characterized. Nucleotide sequence analysis of the reconstructed gene gPP1 revealed a continuous 2430 bp protein coding sequence, with no introns, encoding an 809 amino acid polypeptide. The deduced polypeptide had characteristics of PP1 and contained a 15 amino acid sequence determined by N-terminal peptide sequence analysis of PP1. The sequence of PP1 was highly repetitive with four 200 amino acid sequence domains containing structural motifs in common with cysteine proteinase inhibitors. Expression of the PP1 gene was detected in roots, hypocotyls, cotyledons, stems, and leaves of pumpkin plants. PP1 and its mRNA accumulated in pumpkin hypocotyls during the period of rapid hypocotyl elongation after which mRNA levels declined, while protein levels remained elevated. PP1 was immunolocalized in slime plugs and P-protein bodies in sieve elements of the phloem. Occasionally, PP1 was detected in companion cells. PP1 mRNA was localized by in situ hybridization in companion cells at early stages of vascular differentiation. The developmental accumulation and localization of PP1 and its mRNA paralleled the phloem lactin, further suggesting an interaction between these phloem-specific proteins.

  11. Correlated motion of protein subdomains and large-scale conformational flexibility of RecA protein filament

    Science.gov (United States)

    Yu, Garmay; A, Shvetsov; D, Karelov; D, Lebedev; A, Radulescu; M, Petukhov; V, Isaev-Ivanov

    2012-02-01

    Based on X-ray crystallographic data available at Protein Data Bank, we have built molecular dynamics (MD) models of homologous recombinases RecA from E. coli and D. radiodurans. Functional form of RecA enzyme, which is known to be a long helical filament, was approximated by a trimer, simulated in periodic water box. The MD trajectories were analyzed in terms of large-scale conformational motions that could be detectable by neutron and X-ray scattering techniques. The analysis revealed that large-scale RecA monomer dynamics can be described in terms of relative motions of 7 subdomains. Motion of C-terminal domain was the major contributor to the overall dynamics of protein. Principal component analysis (PCA) of the MD trajectories in the atom coordinate space showed that rotation of C-domain is correlated with the conformational changes in the central domain and N-terminal domain, that forms the monomer-monomer interface. Thus, even though C-terminal domain is relatively far from the interface, its orientation is correlated with large-scale filament conformation. PCA of the trajectories in the main chain dihedral angle coordinate space implicates a co-existence of a several different large-scale conformations of the modeled trimer. In order to clarify the relationship of independent domain orientation with large-scale filament conformation, we have performed analysis of independent domain motion and its implications on the filament geometry.

  12. Swi5-Sfr1 protein stimulates Rad51-mediated DNA strand exchange reaction through organization of DNA bases in the presynaptic filament.

    KAUST Repository

    Fornander, Louise H

    2013-12-03

    The Swi5-Sfr1 heterodimer protein stimulates the Rad51-promoted DNA strand exchange reaction, a crucial step in homologous recombination. To clarify how this accessory protein acts on the strand exchange reaction, we have analyzed how the structure of the primary reaction intermediate, the Rad51/single-stranded DNA (ssDNA) complex filament formed in the presence of ATP, is affected by Swi5-Sfr1. Using flow linear dichroism spectroscopy, we observe that the nucleobases of the ssDNA are more perpendicularly aligned to the filament axis in the presence of Swi5-Sfr1, whereas the bases are more randomly oriented in the absence of Swi5-Sfr1. When using a modified version of the natural protein where the N-terminal part of Sfr1 is deleted, which has no affinity for DNA but maintained ability to stimulate the strand exchange reaction, we still observe the improved perpendicular DNA base orientation. This indicates that Swi5-Sfr1 exerts its activating effect through interaction with the Rad51 filament mainly and not with the DNA. We propose that the role of a coplanar alignment of nucleobases induced by Swi5-Sfr1 in the presynaptic Rad51/ssDNA complex is to facilitate the critical matching with an invading double-stranded DNA, hence stimulating the strand exchange reaction.

  13. Identification of a motor protein required for filamentous growth in Ustilago maydis.

    Science.gov (United States)

    Lehmler, C; Steinberg, G; Snetselaar, K M; Schliwa, M; Kahmann, R; Bölker, M

    1997-06-16

    The phytopathogenic fungus Ustilago maydis exists in two stages, the yeast-like haploid form and the filamentous dikaryon. Both pathogenicity and dimorphism are genetically controlled by two mating-type loci, with only the filamentous stage being pathogenic on corn. We have identified two genes (kin1 and kin2) encoding motor proteins of the kinesin family. Kin1 is most similar to the human CENP-E gene product, while Kin2 is most closely related to the conventional kinesin Nkin of Neurospora crassa. Deletion mutants of kin1 had no discernible phenotype; delta kin2 mutants, however, were severely affected in hyphal extension and pathogenicity. The wild-type dikaryon showed rapid tip growth, with all the cytoplasm being moved to the tip compartment. Left behind are septate cell wall tubes devoid of cytoplasm. In delta kin2 mutants, dikaryotic cells were formed after cell fusion, but these hyphal structures remained short and filled with cytoplasm. A functional green fluorescent protein (GFP)-Kin2 fusion was generated and used to determine the localization of the motor protein by fluorescence microscopy. Inspection of the hyphal tips by electron microscopy revealed a characteristic accumulation of darkly stained vesicles which was absent in mutant cells. We suggest that the motor protein Kin2 is involved in organizing this specialized growth zone at the hyphal tip, probably by affecting the vectorial transport of vesicles.

  14. Protein-Nanocrystal Conjugates Support a Single Filament Polymerization Model in R1 Plasmid Segregation

    Energy Technology Data Exchange (ETDEWEB)

    Choi, Charina L.; Claridge, Shelley A.; Garner, Ethan C.; Alivisatos, A. Paul; Mullins, R. Dyche

    2008-07-15

    To ensure inheritance by daughter cells, many low-copy number bacterial plasmids, including the R1 drug-resistance plasmid, encode their own DNA segregation systems. The par operon of plasmid R1 directs construction of a simple spindle structure that converts free energy of polymerization of an actin-like protein, ParM, into work required to move sister plasmids to opposite poles of rod-shaped cells. The structures of individual components have been solved, but little is known about the ultrastructure of the R1 spindle. To determine the number of ParM filaments in a minimal R1 spindle, we used DNA-gold nanocrystal conjugates as mimics of the R1 plasmid. Wefound that each end of a single polar ParM filament binds to a single ParR/parC-gold complex, consistent with the idea that ParM filaments bind in the hollow core of the ParR/parC ring complex. Our results further suggest that multifilament spindles observed in vivo are associated with clusters of plasmidssegregating as a unit.

  15. Entropic potential field formed for a linear-motor protein near a filament: Statistical-mechanical analyses using simple models

    Science.gov (United States)

    Amano, Ken-ichi; Yoshidome, Takashi; Iwaki, Mitsuhiro; Suzuki, Makoto; Kinoshita, Masahiro

    2010-07-01

    We report a new progress in elucidating the mechanism of the unidirectional movement of a linear-motor protein (e.g., myosin) along a filament (e.g., F-actin). The basic concept emphasized here is that a potential field is entropically formed for the protein on the filament immersed in solvent due to the effect of the translational displacement of solvent molecules. The entropic potential field is strongly dependent on geometric features of the protein and the filament, their overall shapes as well as details of the polyatomic structures. The features and the corresponding field are judiciously adjusted by the binding of adenosine triphosphate (ATP) to the protein, hydrolysis of ATP into adenosine diphosphate (ADP)+Pi, and release of Pi and ADP. As the first step, we propose the following physical picture: The potential field formed along the filament for the protein without the binding of ATP or ADP+Pi to it is largely different from that for the protein with the binding, and the directed movement is realized by repeated switches from one of the fields to the other. To illustrate the picture, we analyze the spatial distribution of the entropic potential between a large solute and a large body using the three-dimensional integral equation theory. The solute is modeled as a large hard sphere. Two model filaments are considered as the body: model 1 is a set of one-dimensionally connected large hard spheres and model 2 is a double helical structure formed by two sets of connected large hard spheres. The solute and the filament are immersed in small hard spheres forming the solvent. The major findings are as follows. The solute is strongly confined within a narrow space in contact with the filament. Within the space there are locations with sharply deep local potential minima along the filament, and the distance between two adjacent locations is equal to the diameter of the large spheres constituting the filament. The potential minima form a ringlike domain in model 1

  16. The nucleoporin Nup98 associates with the intranuclear filamentous protein network of TPR

    Science.gov (United States)

    Fontoura, Beatriz M. A.; Dales, Samuel; Blobel, Günter; Zhong, Hualin

    2001-01-01

    The Nup98 gene codes for several alternatively spliced protein precursors. Two in vitro translated and autoproteolytically cleaved precursors yielded heterodimers of Nup98-6kDa peptide and Nup98-Nup96. TPR (translocated promoter region) is a protein that forms filamentous structures extending from nuclear pore complexes (NPCs) to intranuclear sites. We found that in vitro translated TPR bound to in vitro translated Nup98 and, via Nup98, to Nup96. Double-immunofluorescence microscopy with antibodies to TPR and Nup98 showed colocalization. In confocal sections the nucleolus itself was only weakly stained but there was intensive perinucleolar staining. Striking spike-like structures emanated from this perinucleolar ring and attenuated into thinner structures as they extended to the nuclear periphery. This characteristic staining pattern of the TPR network was considerably enhanced when a myc-tagged pyruvate kinase-6kDa fusion protein was overexpressed in HeLa cells. Double-immunoelectron microscopy of these cells using anti-myc and anti-TPR antibodies and secondary gold-coupled antibodies yielded row-like arrangements of gold particles. Taken together, the immunolocalization data support previous electron microscopical data, suggesting that TPR forms filaments that extend from the NPC to the nucleolus. We discuss the possible implications of the association of Nup98 with this intranuclear TPR network for an intranuclear phase of transport. PMID:11248057

  17. Microbial dynamics and properties of aerobic granules developed in a laboratory-scale sequencing batch reactor with an intermediate filamentous bulking stage.

    Science.gov (United States)

    Aqeel, H; Basuvaraj, M; Hall, M; Neufeld, J D; Liss, S N

    2016-01-01

    Aerobic granules offer enhanced biological nutrient removal and are compact and dense structures resulting in efficient settling properties. Granule instability, however, is still a challenge as understanding of the drivers of instability is poorly understood. In this study, transient instability of aerobic granules, associated with filamentous outgrowth, was observed in laboratory-scale sequencing batch reactors (SBRs). The transient phase was followed by the formation of stable granules. Loosely bound, dispersed, and pinpoint seed flocs gradually turned into granular flocs within 60 days of SBR operation. In stage 1, the granular flocs were compact in structure and typically 0.2 mm in diameter, with excellent settling properties. Filaments appeared and dominated by stage 2, resulting in poor settleability. By stage 3, the SBRs were selected for larger granules and better settling structures, which included filaments that became enmeshed within the granule, eventually forming structures 2-5 mm in diameter. Corresponding changes in sludge volume index were observed that reflected changes in settleability. The protein-to-polysaccharide ratio in the extracted extracellular polymeric substance (EPS) from stage 1 and stage 3 granules was higher (2.8 and 5.7, respectively), as compared to stage 2 filamentous bulking (1.5). Confocal laser scanning microscopic (CLSM) imaging of the biomass samples, coupled with molecule-specific fluorescent staining, confirmed that protein was predominant in stage 1 and stage 3 granules. During stage 2 bulking, there was a decrease in live cells; dead cells predominated. Denaturing gradient gel electrophoresis (DGGE) fingerprint results indicated a shift in bacterial community composition during granulation, which was confirmed by 16S rRNA gene sequencing. In particular, Janthinobacterium (known denitrifier and producer of antimicrobial pigment) and Auxenochlorella protothecoides (mixotrophic green algae) were predominant during stage

  18. Structure of human Rad51 protein filament from molecular modeling and site-specific linear dichroism spectroscopy

    KAUST Repository

    Reymer, A.

    2009-07-08

    To get mechanistic insight into the DNA strand-exchange reaction of homologous recombination, we solved a filament structure of a human Rad51 protein, combining molecular modeling with experimental data. We build our structure on reported structures for central and N-terminal parts of pure (uncomplexed) Rad51 protein by aid of linear dichroism spectroscopy, providing angular orientations of substituted tyrosine residues of Rad51-dsDNA filaments in solution. The structure, validated by comparison with an electron microscopy density map and results from mutation analysis, is proposed to represent an active solution structure of the nucleo-protein complex. An inhomogeneously stretched double-stranded DNA fitted into the filament emphasizes the strategic positioning of 2 putative DNA-binding loops in a way that allows us speculate about their possibly distinct roles in nucleo-protein filament assembly and DNA strand-exchange reaction. The model suggests that the extension of a single-stranded DNA molecule upon binding of Rad51 is ensured by intercalation of Tyr-232 of the L1 loop, which might act as a docking tool, aligning protein monomers along the DNA strand upon filament assembly. Arg-235, also sitting on L1, is in the right position to make electrostatic contact with the phosphate backbone of the other DNA strand. The L2 loop position and its more ordered compact conformation makes us propose that this loop has another role, as a binding site for an incoming double-stranded DNA. Our filament structure and spectroscopic approach open the possibility of analyzing details along the multistep path of the strand-exchange reaction.

  19. Substrate, focal adhesions, and actin filaments: a mechanical unit with a weak spot for mechanosensitive proteins

    International Nuclear Information System (INIS)

    Kirchenbuechler, David; Born, Simone; Kirchgessner, Norbert; Houben, Sebastian; Hoffmann, Bernd; Merkel, Rudolf

    2010-01-01

    Mechanosensing is a vital prerequisite for dynamic remodeling of focal adhesions and cytoskeletal structures upon substrate deformation. For example, tissue formation, directed cell orientation or cell differentiation are regulated by such mechanosensing processes. Focal adhesions and the actin cytoskeleton are believed to be involved in these processes, but where mechanosensing molecules are located and how elastic substrate, focal adhesions and the cytoskeleton couple with each other upon substrate deformation still remains obscure. To approach these questions we have developed a sensitive method to apply defined spatially decaying deformation fields to cells cultivated on ultrasoft elastic substrates and to accurately quantify the resulting displacements of the actin cytoskeleton, focal adhesions, as well as the substrate. Displacement fields were recorded in live cell microscopy by tracking either signals from fluorescent proteins or marker particles in the substrate. As model cell type we used myofibroblasts. These cells are characterized by highly stable adhesion and force generating structures but are still able to detect mechanical signals with high sensitivity. We found a rigid connection between substrate and focal adhesions. Furthermore, stress fibers were found to be barely extendable almost over their whole lengths. Plastic deformation took place only at the very ends of actin filaments close to focal adhesions. As a result, this area became elongated without extension of existing actin filaments by polymerization. Both ends of the stress fibers were mechanically coupled with detectable plastic deformations on either site. Interestingly, traction force dependent substrate deformation fields remained mostly unaffected even when stress fiber elongations were released. These data argue for a location of mechanosensing proteins at the ends of actin stress fibers and describe, except for these domains, the whole system to be relatively rigid for tensile

  20. Proteomics characterization of intermediate wheatgrass (Thinopyrum intermedium) flour proteins

    Science.gov (United States)

    Thinopyrum intermedium, commonly known as intermediate wheatgrass (IWG), is a perennial crop with favorable agronomic characteristics and nutritional benefits. IWG lines are deficient in high molecular weight glutenins (HMWG), responsible for dough strength. A detailed characterization of IWG flou...

  1. Intermediate filaments of zebrafish retinal and optic nerve astrocytes and M?ller glia: differential distribution of cytokeratin and GFAP

    OpenAIRE

    Koke, Joseph R; Mosier, Amanda L; Garc?a, Dana M

    2010-01-01

    Abstract Background Optic nerve regeneration (ONR) following injury is a model for central nervous system regeneration. In zebrafish, ONR is rapid - neurites cross the lesion and enter the optic tectum within 7 days; in mammals regeneration does not take place unless astrocytic reactivity is suppressed. Glial fibrillary acidic protein (GFAP) is used as a marker for retinal and optic nerve astrocytes in both fish and mammals, even though it has long been known that astrocytes of optic nerves i...

  2. F-actin-like filaments formed by plasmid segregation protein ParM

    DEFF Research Database (Denmark)

    van den Ent, Fusinita; Møller-Jensen, Jakob; Amos, Linda A.

    2002-01-01

    It was the general belief that DNA partitioning in prokaryotes is independent of a cytoskeletal structure, which in eukaryotic cells is indispensable for DNA segregation. Recently, however, immunofluorescence microscopy revealed highly dynamic, filamentous structures along the longitudinal axis o...... compared with F-actin, despite the similar arrangement of the subunits within the filaments. Thus, there is now evidence for cytoskeletal structures, formed by actin-like filaments that are involved in plasmid partitioning in E.coli. Udgivelsesdato: Dec 16...

  3. The Molecular Architecture for the Intermediate Filaments of Hard [alpha]-Keratin Based on the Superlattice Data Obtained from a Study ofMammals Using Synchrotron Fibre Diffraction

    Energy Technology Data Exchange (ETDEWEB)

    James, Veronica (ANU)

    2014-09-24

    High- and low-angle X-ray diffraction studies of hard {alpha}-keratin have been studied, and various models have been proposed over the last 70 years. Most of these studies have been confined to one or two forms of alpha keratin. This high- and low-angle synchrotron fibre diffraction study extends the study to cover all available data for all known forms of hard {alpha}-keratin including hairs, fingernails, hooves, horn, and quills from mammals, marsupials, and a monotreme, and it confirms that the model proposed is universally acceptable for all mammals. A complete Bragg analysis of the meridional diffraction patterns, including multiple-time exposures to verify any weak reflections, verified the existence of a superlattice consisting of two infinite lattices and three finite lattices. An analysis of the equatorial patterns establishes the radii of the oligomeric levels of dimers, tetramers, and intermediate filaments (IFs) together with the centre to centre distance for the IFs, thus confirming the proposed helices within helices molecular architecture for hard {alpha}-keratin. The results verify that the structure proposed by Feughelman and James meets the criteria for a valid {alpha}-keratin structure.

  4. The importance of connections between the cell wall integrity pathway and the unfolded protein response in filamentous fungi.

    Science.gov (United States)

    Malavazi, Iran; Goldman, Gustavo Henrique; Brown, Neil Andrew

    2014-11-01

    In the external environment, or within a host organism, filamentous fungi experience sudden changes in nutrient availability, osmolality, pH, temperature and the exposure to toxic compounds. The fungal cell wall represents the first line of defense, while also performing essential roles in morphology, development and virulence. A polarized secretion system is paramount for cell wall biosynthesis, filamentous growth, nutrient acquisition and interactions with the environment. The unique ability of filamentous fungi to secrete has resulted in their industrial adoption as fungal cell factories. Protein maturation and secretion commences in the endoplasmic reticulum (ER). The unfolded protein response (UPR) maintains ER functionality during exposure to secretion and cell wall stress. UPR, therefore, influences secretion and cell wall homeostasis, which in turn impacts upon numerous fungal traits important to pathogenesis and biotechnology. Subsequently, this review describes the relevance of the cell wall and UPR systems to filamentous fungal pathogens or industrial microbes and then highlights interconnections between the two systems. Ultimately, the possible biotechnological applications of an enhanced understanding of such regulatory systems in combating fungal disease, or the removal of natural bottlenecks in protein secretion in an industrial setting, are discussed. © The Author 2014. Published by Oxford University Press. All rights reserved. For permissions, please email: journals.permissions@oup.com.

  5. Engineering the Controlled Assembly of Filamentous Injectisomes in E. coli K-12 for Protein Translocation into Mammalian Cells.

    Science.gov (United States)

    Ruano-Gallego, David; Álvarez, Beatriz; Fernández, Luis Ángel

    2015-09-18

    Bacterial pathogens containing type III protein secretion systems (T3SS) assemble large needle-like protein complexes in the bacterial envelope, called injectisomes, for translocation of protein effectors into host cells. The application of these "molecular syringes" for the injection of proteins into mammalian cells is hindered by their structural and genomic complexity, requiring multiple polypeptides encoded along with effectors in various transcriptional units (TUs) with intricate regulation. In this work, we have rationally designed the controlled expression of the filamentous injectisomes found in enteropathogenic Escherichia coli (EPEC) in the nonpathogenic strain E. coli K-12. All structural components of EPEC injectisomes, encoded in a genomic island called the locus of enterocyte effacement (LEE), were engineered in five TUs (eLEEs) excluding effectors, promoters and transcriptional regulators. These eLEEs were placed under the control of the IPTG-inducible promoter Ptac and integrated into specific chromosomal sites of E. coli K-12 using a marker-less strategy. The resulting strain, named synthetic injector E. coli (SIEC), assembles filamentous injectisomes similar to those in EPEC. SIEC injectisomes form pores in the host plasma membrane and are able to translocate T3-substrate proteins (e.g., translocated intimin receptor, Tir) into the cytoplasm of HeLa cells reproducing the phenotypes of intimate attachment and polymerization of actin-pedestals elicited by EPEC bacteria. Hence, SIEC strain allows the controlled expression of functional filamentous injectisomes for efficient translocation of proteins with T3S-signals into mammalian cells.

  6. Why do alpha-beta parallel proteins, like flavodoxins, form misfolded off-pathway intermediates?

    NARCIS (Netherlands)

    Nabuurs, S.M.

    2009-01-01

    The question: “Why do α-β parallel proteins, like flavodoxins, form misfolded off-pathway
    intermediates?" is the main subject of this thesis. A. vinelandii apoflavodoxin is chosen as protein
    of interest as it is a representative of α-β parallel proteins, which are widely prevalent in

  7. Structural and Functional Studies of H. seropedicae RecA Protein – Insights into the Polymerization of RecA Protein as Nucleoprotein Filament

    Energy Technology Data Exchange (ETDEWEB)

    Leite, Wellington C.; Galvão, Carolina W.; Saab, Sérgio C.; Iulek, Jorge; Etto, Rafael M.; Steffens, Maria B.R.; Chitteni-Pattu, Sindhu; Stanage, Tyler; Keck, James L.; Cox, Michael M. (UW); (UW-MED); (Ponta Grossa)

    2016-07-22

    The bacterial RecA protein plays a role in the complex system of DNA damage repair. Here, we report the functional and structural characterization of the Herbaspirillum seropedicae RecA protein (HsRecA). HsRecA protein is more efficient at displacing SSB protein from ssDNA than Escherichia coli RecA protein. HsRecA also promotes DNA strand exchange more efficiently. The three dimensional structure of HsRecA-ADP/ATP complex has been solved to 1.7 Å resolution. HsRecA protein contains a small N-terminal domain, a central core ATPase domain and a large C-terminal domain, that are similar to homologous bacterial RecA proteins. Comparative structural analysis showed that the N-terminal polymerization motif of archaeal and eukaryotic RecA family proteins are also present in bacterial RecAs. Reconstruction of electrostatic potential from the hexameric structure of HsRecA-ADP/ATP revealed a high positive charge along the inner side, where ssDNA is bound inside the filament. The properties of this surface may explain the greater capacity of HsRecA protein to bind ssDNA, forming a contiguous nucleoprotein filament, displace SSB and promote DNA exchange relative to EcRecA. In conclusion, our functional and structural analyses provide insight into the molecular mechanisms of polymerization of bacterial RecA as a helical nucleoprotein filament.

  8. Identification of intermediate species in protein-folding by ...

    Indian Academy of Sciences (India)

    TECS

    mixtures of non-interacting molecules such as fluorescent dyes, peptides and proteins. ... state, it is necessary to observe parameters which are sensitive to the ... them. Thus, such measurements offer the possibility of monitoring various regions of a protein during the transition from the U to the N form. When coupled with the ...

  9. Multi-subunit proteins on the surface of filamentous phage: methodologies for displaying antibody (Fab) heavy and light chains.

    OpenAIRE

    Hoogenboom, H R; Griffiths, A D; Johnson, K S; Chiswell, D J; Hudson, P; Winter, G

    1991-01-01

    The display of proteins on the surface of phage offers a powerful means of selecting for rare genes encoding proteins with binding activities. Recently we found that antibody heavy and light chain variable (V) domains fused as a single polypeptide chain to a minor coat protein of filamentous phage fd, could be enriched by successive rounds of phage growth and panning with antigen. This allows the selection of antigen-binding domains directly from diverse libraries of V-genes. Now we show that...

  10. Cloning and sequencing of the cDNA encoding a core protein of the paired helical filament of Alzheimer's disease: Identification as the microtubule-associated protein tau

    International Nuclear Information System (INIS)

    Goedert, M.; Wischik, C.M.; Crowther, R.A.; Walker, J.E.; Klug, A.

    1988-01-01

    Screening of cDNA libraries prepared from the frontal cortex of an Alzheimer's disease patient and from fetal human brain has led to isolation of the cDNA for a core protein of the paired helical filament of Alzheimer's disease. The partial amino acid sequence of this core protein was used to design synthetic oligonucleotide probes. The cDNA encodes a protein of 352 amino acids that contains a characteristic amino acid repeat in its carboxyl-terminal half. This protein is highly homologous to the sequence of the mouse microtubule-associated protein tau and thus constitutes the human equivalent of mouse tau. RNA blot analysis indicates the presence of two major transcripts, 6 and 2 kilobases long, with a wide distribution in normal human brain. Tau protein mRNAs were found in normal amounts in the frontal cortex from patients with Alzheimer's disease. The proof that at least part of tau protein forms a component of the paired helical filament core opens the way to understanding the mode of formation of paired helical filaments and thus, ultimately, the pathogenesis of Alzheimer's disease

  11. Identification of intermediate species in protein-folding by ...

    Indian Academy of Sciences (India)

    TECS

    of monitoring various regions of a protein during the transition from the U to the N form. When coupled with the maximum entropy method (MEM) of analysis of intensity decay curves,. 1,2. TR-FRET measurements can resolve heterogeneity in terms of distributions of intramolecular distances. Much has been learnt about.

  12. The Transmembrane Morphogenesis Protein gp1 of Filamentous Phages Contains Walker A and Walker B Motifs Essential for Phage Assembly

    Directory of Open Access Journals (Sweden)

    Belinda Loh

    2017-04-01

    Full Text Available In contrast to lytic phages, filamentous phages are assembled in the inner membrane and secreted across the bacterial envelope without killing the host. For assembly and extrusion of the phage across the host cell wall, filamentous phages code for membrane-embedded morphogenesis proteins. In the outer membrane of Escherichia coli, the protein gp4 forms a pore-like structure, while gp1 and gp11 form a complex in the inner membrane of the host. By comparing sequences with other filamentous phages, we identified putative Walker A and B motifs in gp1 with a conserved lysine in the Walker A motif (K14, and a glutamic and aspartic acid in the Walker B motif (D88, E89. In this work we demonstrate that both, Walker A and Walker B, are essential for phage production. The crucial role of these key residues suggests that gp1 might be a molecular motor driving phage assembly. We further identified essential residues for the function of the assembly complex. Mutations in three out of six cysteine residues abolish phage production. Similarly, two out of six conserved glycine residues are crucial for gp1 function. We hypothesise that the residues represent molecular hinges allowing domain movement for nucleotide binding and phage assembly.

  13. The Transmembrane Morphogenesis Protein gp1 of Filamentous Phages Contains Walker A and Walker B Motifs Essential for Phage Assembly.

    Science.gov (United States)

    Loh, Belinda; Haase, Maximilian; Mueller, Lukas; Kuhn, Andreas; Leptihn, Sebastian

    2017-04-09

    In contrast to lytic phages, filamentous phages are assembled in the inner membrane and secreted across the bacterial envelope without killing the host. For assembly and extrusion of the phage across the host cell wall, filamentous phages code for membrane-embedded morphogenesis proteins. In the outer membrane of Escherichia coli, the protein gp4 forms a pore-like structure, while gp1 and gp11 form a complex in the inner membrane of the host. By comparing sequences with other filamentous phages, we identified putative Walker A and B motifs in gp1 with a conserved lysine in the Walker A motif (K14), and a glutamic and aspartic acid in the Walker B motif (D88, E89). In this work we demonstrate that both, Walker A and Walker B, are essential for phage production. The crucial role of these key residues suggests that gp1 might be a molecular motor driving phage assembly. We further identified essential residues for the function of the assembly complex. Mutations in three out of six cysteine residues abolish phage production. Similarly, two out of six conserved glycine residues are crucial for gp1 function. We hypothesise that the residues represent molecular hinges allowing domain movement for nucleotide binding and phage assembly.

  14. The Bipolar Filaments Formed by Herpes Simplex Virus Type 1 SSB/Recombination Protein (ICP8) Suggest a Mechanism for DNA Annealing

    Energy Technology Data Exchange (ETDEWEB)

    Makhov, A.M.; Simon, M.; Sen, A.; Yu, X.; Griffith, J. D.; Egelman, E. H.

    2009-02-20

    Herpes simplex virus type 1 encodes a multifunctional protein, ICP8, which serves both as a single-strand binding protein and as a recombinase, catalyzing reactions involved in replication and recombination of the viral genome. In the presence of divalent ions and at low temperature, previous electron microscopic studies showed that ICP8 will form long left-handed helical filaments. Here, electron microscopic image reconstruction reveals that the filaments are bipolar, with an asymmetric unit containing two subunits of ICP8 that constitute a symmetrical dimer. This organization of the filament has been confirmed using scanning transmission electron microscopy. The pitch of the filaments is {approx} 250 {angstrom}, with {approx} 6.2 dimers per turn. Docking of a crystal structure of ICP8 into the reconstructed filament shows that the C-terminal domain of ICP8, attached to the body of the subunit by a flexible linker containing {approx} 10 residues, is packed into a pocket in the body of a neighboring subunit in the crystal in a similar manner as in the filament. However, the interactions between the large N-terminal domains are quite different in the filament from that observed in the crystal. A previously proposed model for ICP8 binding single-stranded DNA (ssDNA), based upon the crystal structure, leads to a model for a continuous strand of ssDNA near the filament axis. The bipolar nature of the ICP8 filaments means that a second strand of ssDNA would be running through this filament in the opposite orientation, and this provides a potential mechanism for how ICP8 anneals complementary ssDNA into double-stranded DNA, where each strand runs in opposite directions.

  15. Expressão dos filamentos intermediários no diagnóstico dos tumores mamários de cadelas Expression of intermediate filaments in canine mammary tumors diagnosis

    Directory of Open Access Journals (Sweden)

    D.A.P.C. Zuccari

    2002-12-01

    Full Text Available Foram utilizados anticorpos monoclonais para marcação imunoistoquímica dos tecidos tumorais e obtenção de informações sobre a histogênese dos tumores mamários utilizando-se anti-citoqueratinas para marcação de células epiteliais, e anti-actina e anti-vimentina para células mioepiteliais. O procedimento imunoistoquímico mostrou-se esclarecedor com relação à histogênese dos tumores mamários, confirmando a marcação de células epiteliais com as citoqueratinas que perdem sua expressão na transformação celular maligna. A alfa-actina e a vimentina mostraram-se eficientes na marcação de células mioepiteliais. A alfa-actina diminuiu a marcação na metaplasia óssea ou cartilaginosa contrariamente à vimentina cuja marcação foi aumentada. Os resultados permitem melhor entendimento da classificação dos tumores mamários de cadelas com a utilização de anticorpos monoclonais como marcadores do citoesqueleto, que se mostraram eficientes nessa caracterização.Immunohistochemical evaluation was performed to study the histogenesis of canine mammary tumors and to contribute to a better understanding of their classification. Monoclonal antibodies specific for different types of intermediate filaments (cytokeratins, vimentin, alpha-actin were used. Epithelial cells stained positively for cytokeratins and their expression was lost as the malignant transformation occurs. Myoepithelial cells stained positively for vimentin and alpha-actin. In contrast to vimentin, alpha-actin lost the expression as the cartilaginous or osseous metaplasia occurs. Immunohistochemical evaluation with monoclonal antibodies proved to be efficient for identification of tumor histogenesis. alpha-actin were used. Epithelial cells stained positively for cytokeratins and their expression was lost as the malignant transformation occurs. Myoepithelial cells stained positively for vimentin and alpha-actin. In contrast to vimentin, alpha-actin lost the expression

  16. A Robust Actin Filaments Image Analysis Framework.

    Directory of Open Access Journals (Sweden)

    Mitchel Alioscha-Perez

    2016-08-01

    Full Text Available The cytoskeleton is a highly dynamical protein network that plays a central role in numerous cellular physiological processes, and is traditionally divided into three components according to its chemical composition, i.e. actin, tubulin and intermediate filament cytoskeletons. Understanding the cytoskeleton dynamics is of prime importance to unveil mechanisms involved in cell adaptation to any stress type. Fluorescence imaging of cytoskeleton structures allows analyzing the impact of mechanical stimulation in the cytoskeleton, but it also imposes additional challenges in the image processing stage, such as the presence of imaging-related artifacts and heavy blurring introduced by (high-throughput automated scans. However, although there exists a considerable number of image-based analytical tools to address the image processing and analysis, most of them are unfit to cope with the aforementioned challenges. Filamentous structures in images can be considered as a piecewise composition of quasi-straight segments (at least in some finer or coarser scale. Based on this observation, we propose a three-steps actin filaments extraction methodology: (i first the input image is decomposed into a 'cartoon' part corresponding to the filament structures in the image, and a noise/texture part, (ii on the 'cartoon' image, we apply a multi-scale line detector coupled with a (iii quasi-straight filaments merging algorithm for fiber extraction. The proposed robust actin filaments image analysis framework allows extracting individual filaments in the presence of noise, artifacts and heavy blurring. Moreover, it provides numerous parameters such as filaments orientation, position and length, useful for further analysis. Cell image decomposition is relatively under-exploited in biological images processing, and our study shows the benefits it provides when addressing such tasks. Experimental validation was conducted using publicly available datasets, and in

  17. Functional display of proteins, mutant proteins, fragments of proteins and peptides on the surface of filamentous (bacterio) phages: A review

    NARCIS (Netherlands)

    Pannekoek, H.; van Meijer, M.; Gaardsvoll, H.; van Zonneveld, A. J.

    1995-01-01

    Cytoplasmic expression of complex eukaryotic proteins inEscherichia coli usually yields inactive protein preparations. In some cases, (part) of the biological activity can be recovered by rather inefficient denaturation-renaturation procedures. Recently, novel concepts have been developed for the

  18. Probing the Energetics of Dynactin Filament Assembly and the Binding of Cargo Adaptor Proteins Using Molecular Dynamics Simulation and Electrostatics-Based Structural Modeling.

    Science.gov (United States)

    Zheng, Wenjun

    2017-01-10

    Dynactin, a large multiprotein complex, binds with the cytoplasmic dynein-1 motor and various adaptor proteins to allow recruitment and transportation of cellular cargoes toward the minus end of microtubules. The structure of the dynactin complex is built around an actin-like minifilament with a defined length, which has been visualized in a high-resolution structure of the dynactin filament determined by cryo-electron microscopy (cryo-EM). To understand the energetic basis of dynactin filament assembly, we used molecular dynamics simulation to probe the intersubunit interactions among the actin-like proteins, various capping proteins, and four extended regions of the dynactin shoulder. Our simulations revealed stronger intersubunit interactions at the barbed and pointed ends of the filament and involving the extended regions (compared with the interactions within the filament), which may energetically drive filament termination by the capping proteins and recruitment of the actin-like proteins by the extended regions, two key features of the dynactin filament assembly process. Next, we modeled the unknown binding configuration among dynactin, dynein tails, and a number of coiled-coil adaptor proteins (including several Bicaudal-D and related proteins and three HOOK proteins), and predicted a key set of charged residues involved in their electrostatic interactions. Our modeling is consistent with previous findings of conserved regions, functional sites, and disease mutations in the adaptor proteins and will provide a structural framework for future functional and mutational studies of these adaptor proteins. In sum, this study yielded rich structural and energetic information about dynactin and associated adaptor proteins that cannot be directly obtained from the cryo-EM structures with limited resolutions.

  19. Glutamate Induced Thermal Equilibrium Intermediate and Counteracting Effect on Chemical Denaturation of Proteins.

    Science.gov (United States)

    Anumalla, Bramhini; Prabhu, N Prakash

    2018-01-25

    When organisms are subjected to stress conditions, one of their adaptive responses is accumulation of small organic molecules called osmolytes. These osmolytes affect the structure and stability of the biological macromolecules including proteins. The present study examines the effect of a negatively charged amino acid osmolyte, glutamate (Glu), on two model proteins, ribonuclease A (RNase A) and α-lactalbumin (α-LA), which have positive and negative surface charges at pH 7, respectively. These proteins follow two-state unfolding transitions during both heat and chemical induced denaturation processes. The addition of Glu stabilizes the proteins against temperature and induces an early equilibrium intermediate during unfolding. The stability is found to be enthalpy-driven, and the free energy of stabilization is more for α-LA compared to RNase A. The decrease in the partial molar volume and compressibility of both of the proteins in the presence of Glu suggests that the proteins attain a more compact state through surface hydration which could provide a more stable conformation. This is also supported by molecule dynamic simulation studies which demonstrate that the water density around the proteins is increased upon the addition of Glu. Further, the intermediates could be completely destabilized by lower concentrations (∼0.5 M) of guanidinium chloride and salt. However, urea subverts the Glu-induced intermediate formed by α-LA, whereas it only slightly destabilizes in the case of RNase A which has a positive surface charge and could possess charge-charge interactions with Glu. This suggests that, apart from hydration, columbic interactions might also contribute to the stability of the intermediate. Gdm-induced denaturation of RNase A and α-LA in the absence and the presence of Glu at different temperatures was carried out. These results also show the Glu-induced stabilization of both of the proteins; however, all of the unfolding transitions followed two

  20. Enhancement of DNA vaccine potency through linkage of antigen to filamentous bacteriophage coat protein III domain I

    DEFF Research Database (Denmark)

    Cuesta, Àngel M; Suárez, Eduardo; Larsen, Martin

    2006-01-01

    Although DNA-based cancer vaccines have been successfully tested in mouse models, a major drawback of cancer vaccination still remains, namely that tumour antigens are weak and fail to generate a vigorous immune response in tumour-bearing patients. Genetic technology offers strategies for promoting...... immune pathways by adding immune-activating genes to the tumour antigen sequence. In this work, we converted a model non-immunogenic antigen into a vaccine by fusing it to domain I of the filamentous bacteriophage coat protein III gene. Vaccination with a DNA construct encoding the domain I fusion...... generated antigen-specific T helper 1-type cellular immune responses. These results demonstrate that the incorporation of protein III into a DNA vaccine formulation can modulate the gene-mediated immune response and may thus provide a strategy for improving its therapeutic effect....

  1. The ribosome can prevent aggregation of partially folded protein intermediates: studies using the Escherichia coli ribosome.

    Directory of Open Access Journals (Sweden)

    Bani Kumar Pathak

    Full Text Available BACKGROUND: Molecular chaperones that support de novo folding of proteins under non stress condition are classified as chaperone 'foldases' that are distinct from chaperone' holdases' that provide high affinity binding platform for unfolded proteins and prevent their aggregation specifically under stress conditions. Ribosome, the cellular protein synthesis machine can act as a foldase chaperone that can bind unfolded proteins and release them in folding competent state. The peptidyl transferase center (PTC located in the domain V of the 23S rRNA of Escherichia coli ribosome (bDV RNA is the chaperoning center of the ribosome. It has been proposed that via specific interactions between the RNA and refolding proteins, the chaperone provides information for the correct folding of unfolded polypeptide chains. RESULTS: We demonstrate using Escherichia coli ribosome and variants of its domain V RNA that the ribosome can bind to partially folded intermediates of bovine carbonic anhydrase II (BCAII and lysozyme and suppress aggregation during their refolding. Using mutants of domain V RNA we demonstrate that the time for which the chaperone retains the bound protein is an important factor in determining its ability to suppress aggregation and/or support reactivation of protein. CONCLUSION: The ribosome can behave like a 'holdase' chaperone and has the ability to bind and hold back partially folded intermediate states of proteins from participating in the aggregation process. Since the ribosome is an essential organelle that is present in large numbers in all living cells, this ability of the ribosome provides an energetically inexpensive way to suppress cellular aggregation. Further, this ability of the ribosome might also be crucial in the context that the ribosome is one of the first chaperones to be encountered by a large nascent polypeptide chains that have a tendency to form partially folded intermediates immediately following their synthesis.

  2. Capping protein binding to S100B: implications for the tentacle model for capping the actin filament barbed end.

    Science.gov (United States)

    Wear, Martin A; Cooper, John A

    2004-04-02

    S100B binds tightly to a 12-amino acid peptide derived from heterodimeric capping protein. In native intact capping protein, this sequence is in the C terminus of the alpha-subunit, which is important for capping the actin filament. This C-terminal region is proposed to act as a flexible "tentacle," extending away from the body of capping protein in order to bind actin. To this hypothesis, we analyzed the interaction between S100B and capping protein in solution. The C-terminal 28 amino acids of the alpha-subunit, the proposed tentacle, bound to S100B as a free synthetic peptide or a glutathione S-transferase fusion (K(d) approximately 0.4-1 microm). In contrast, S100B did not bind to whole native capping protein or functionally affect its capping activity. S100B does not bind, with any significant affinity, to the proposed alpha-tentacle sequence of whole native capping protein in solution. In the NMR structure of S100B complexed with the alpha-subunit-derived 12-amino acid peptide, the hydrophobic side of a short alpha-helix in the peptide, containing an important tryptophan residue, contacts S100B. In the x-ray structure of native capping protein, the corresponding sequence of the alpha-subunit C terminus, including Trp(271), interacts closely with the body of the protein. Therefore, our results suggest the alpha-subunit C terminus is not mobile as predicted by the tentacle model. Addition of non-ionic detergent allowed whole capping protein to bind weakly to S100B, indicating that the alpha-subunit C terminus can be mobilized from the surface of the capping protein molecule, presumably by weakening the hydrophobic binding at the contact site.

  3. Protein-segment universe exhibiting transitions at intermediate segment length in conformational subspaces

    Directory of Open Access Journals (Sweden)

    Hirokawa Takatsugu

    2008-08-01

    Full Text Available Abstract Background Many studies have examined rules governing two aspects of protein structures: short segments and proteins' structural domains. Nevertheless, the organization and nature of the conformational space of segments with intermediate length between short segments and domains remain unclear. Conformational spaces of intermediate length segments probably differ from those of short segments. We investigated the identification and characterization of the boundary(s between peptide-like (short segment and protein-like (long segment distributions. We generated ensembles embedded in globular proteins comprising segments 10–50 residues long. We explored the relationships between the conformational distribution of segments and their lengths, and also protein structural classes using principal component analysis based on the intra-segment Cα-Cα atomic distances. Results Our statistical analyses of segment conformations and length revealed critical dual transitions in their conformational distribution with segments derived from all four structural classes. Dual transitions were identified with the intermediate phase between the short segments and domains. Consequently, protein segment universes were categorized. i Short segments (10–22 residues showed a distribution with a high frequency of secondary structure clusters. ii Medium segments (23���26 residues showed a distribution corresponding to an intermediate state of transitions. iii Long segments (27–50 residues showed a distribution converging on one huge cluster containing compact conformations with a smaller radius of gyration. This distribution reflects the protein structures' organization and protein domains' origin. Three major conformational components (radius of gyration, structural symmetry with respect to the N-terminal and C-terminal halves, and single-turn/two-turn structure well define most of the segment universes. Furthermore, we identified several

  4. Nucleotide cofactor-dependent structural change of Xenopus laevis Rad51 protein filament detected by small-angle neutron scattering measurements in solution

    DEFF Research Database (Denmark)

    Ellouze, C.; Kim, H.-K.; Maeshima, K.

    1997-01-01

    of RecA. Upon ATP binding under high-salt conditions (600 mM NaCl), the helical pitch of XRad51.1 filament was increased from 8 to 10 nm and the cross-sectional diameter decreased from 7 to 6 nm. The pitch sizes of XRad51.1 are similar to, though slightly larger than, those of RecA filament under......Rad51 protein, a eukaryotic homologue of RecA protein, forms a filamentous complex with DNA and catalyzes homologous recombination. We have analyzed the structure of Xenopus Rad51 protein (XRad51.1) in solution by small-angle neutron scattering (SANS). The measurements showed that XRad51.1 forms...... a helical filament independently of DNA. The sizes of the cross-sectional and helical pitch of the filament could be determined, respectively, from a Guinier plot and the position of the subsidiary maximum of SANS data. We observed that the helical structure is modified by nucleotide binding as in the case...

  5. Mapping and Identification of Antifungal Peptides in the Putative Antifungal Protein AfpB from the Filamentous Fungus Penicillium digitatum.

    Science.gov (United States)

    Garrigues, Sandra; Gandía, Mónica; Borics, Attila; Marx, Florentine; Manzanares, Paloma; Marcos, Jose F

    2017-01-01

    Antifungal proteins (AFPs) from Ascomycetes are small cysteine-rich proteins that are abundantly secreted and show antifungal activity against non-producer fungi. A gene coding for a class B AFP (AfpB) was previously identified in the genome of the plant pathogen Penicillium digitatum . However, previous attempts to detect the AfpB protein were not successful despite the high expression of the corresponding afpB gene. In this work, the structure of the putative AfpB was modeled. Based on this model, four synthetic cysteine-containing peptides, PAF109, PAF112, PAF118, and PAF119, were designed and their antimicrobial activity was tested and characterized. PAF109 that corresponds to the γ-core motif present in defensin-like antimicrobial proteins did not show antimicrobial activity. On the contrary, PAF112 and PAF118, which are cationic peptides derived from two surface-exposed loops in AfpB, showed moderate antifungal activity against P. digitatum and other filamentous fungi. It was also confirmed that cyclization through a disulfide bridge prevented peptide degradation. PAF116, which is a peptide analogous to PAF112 but derived from the Penicillium chrysogenum antifungal protein PAF, showed activity against P. digitatum similar to PAF112, but was less active than the native PAF protein. The two AfpB-derived antifungal peptides PAF112 and PAF118 showed positive synergistic interaction when combined against P. digitatum . Furthermore, the synthetic hexapeptide PAF26 previously described in our laboratory also exhibited synergistic interaction with the peptides PAF112, PAF118, and PAF116, as well as with the PAF protein. This study is an important contribution to the mapping of antifungal motifs within the AfpB and other AFPs, and opens up new strategies for the rational design and application of antifungal peptides and proteins.

  6. Determination of Protein Folding Intermediate Structures Consistent with Data from Oxidative Footprinting Mass Spectrometry.

    Science.gov (United States)

    Heinkel, Florian; Gsponer, Jörg

    2016-01-29

    The mapping of folding landscapes remains an important challenge in protein chemistry. Pulsed oxidative labeling of exposed residues and their detection via mass spectrometry provide new means of taking time-resolved "snapshots" of the structural changes that occur during protein folding. However, such experiments have been so far only interpreted qualitatively. Here, we report the detailed structural interpretation of mass spectrometry data from fast photochemical oxidation of proteins (FPOP) experiments at atomic resolution in a biased molecular dynamics approach. We are able to calculate structures of the early folding intermediate of the model system barstar that are fully consistent with FPOP data and Φ values. Furthermore, structures calculated with both FPOP data and Φ values are significantly less compact and have fewer helical residues than intermediate structures calculated with Φ values only. This improves the agreement with the experimental β-Tanford value and CD measurements. The restraints that we introduce facilitate the structural interpretation of FPOP data and provide new means for refined structure calculations of transiently sampled states on protein folding landscapes. Copyright © 2015 Elsevier Ltd. All rights reserved.

  7. Occurrence of proteinaceous 10-nm filaments throughout the cytoplasm of algae of the order Dasycladales.

    Science.gov (United States)

    Berger, S; Wittke, W; Traub, P

    1998-05-01

    Previously, whole-mount electron microscopy of nuclei extruded together with residual cytoplasm from the rhizoids of several algal species of the order Dasycladales has revealed the occurrence of an intra- and perinuclear network of 10-nm filaments morphologically indistinguishable from that of mammalian vimentin intermediate filaments. The present investigation demonstrates the existence of a filament system throughout the cytoplasm of the rhizoid, stalk, and apical tip of these giant cells. However, while the perinuclear 10-nm filaments interconnecting the nuclear surface with a perinuclear layer of large, electron-dense bodies filled with nucleoprotein material are of smooth appearance, those continuing within and beyond the perinuclear bodies are densely covered with differently sized, globular structures and, therefore, are of a very rough appearance. The filaments in the very apical tip of the cells are mainly of the smooth type. The transition from smooth to rough filaments seems to occur in the numerous perinuclear dense bodies surrounding the large nucleus. Digestion of the rough filaments with proteinase K removes the globules from the filament surface, revealing that throughout the nonvacuolar, intracellular space the filaments have the same basic 10-nm structure. On the other hand, gold-conjugated RNase A strongly binds to the filament-attached globules but not to the smooth, perinuclear, and the proteinase K-treated, rough filaments. In addition, an antibody raised against Xp54, a highly conserved protein which in Xenopus oocytes is an integral component of stored mRNP particles, decorates the rough but not the smooth 10-nm filaments. These results support the notion that the 10-nm filament system of Dasycladales cells plays a role in the transient storage of ribonucleoprotein particles in the cytoplasm and possibly fulfils a supportive function in the actomyosin-based transport of such material to various cytological destinations.

  8. DNA Binding Proteins of the Filamentous Phages CTXφ and VGJφ of Vibrio cholerae▿

    Science.gov (United States)

    Falero, Alina; Caballero, Andy; Ferrán, Beatriz; Izquierdo, Yovanny; Fando, Rafael; Campos, Javier

    2009-01-01

    The native product of open reading frame 112 (orf112) and a recombinant variant of the RstB protein, encoded by Vibrio cholerae pathogen-specific bacteriophages VGJφ and CTXφ, respectively, were purified to more than 90% homogeneity. Orf112 protein was shown to specifically bind single-stranded genomic DNA of VGJφ; however, RstB protein unexpectedly bound double-stranded DNA in addition to the single-stranded genomic DNA. The DNA binding properties of these proteins may explain their requirement for the rolling circle replication of the respective phages and RstB's requirement for single-stranded-DNA chromosomal integration of CTXφ phage dependent on XerCD recombinases. PMID:19617366

  9. DNA binding proteins of the filamentous phages CTXphi and VGJphi of Vibrio cholerae.

    Science.gov (United States)

    Falero, Alina; Caballero, Andy; Ferrán, Beatriz; Izquierdo, Yovanny; Fando, Rafael; Campos, Javier

    2009-09-01

    The native product of open reading frame 112 (orf112) and a recombinant variant of the RstB protein, encoded by Vibrio cholerae pathogen-specific bacteriophages VGJphi and CTXphi, respectively, were purified to more than 90% homogeneity. Orf112 protein was shown to specifically bind single-stranded genomic DNA of VGJphi; however, RstB protein unexpectedly bound double-stranded DNA in addition to the single-stranded genomic DNA. The DNA binding properties of these proteins may explain their requirement for the rolling circle replication of the respective phages and RstB's requirement for single-stranded-DNA chromosomal integration of CTXphi phage dependent on XerCD recombinases.

  10. Antibody-based analysis reveals “filamentous vs. non-filamentous” and “cytoplasmic vs. nuclear” crosstalk of cytoskeletal proteins

    Energy Technology Data Exchange (ETDEWEB)

    Kumeta, Masahiro, E-mail: kumeta@lif.kyoto-u.ac.jp [Graduate School of Biostudies, Kyoto University, Kyoto 606-8501 (Japan); Hirai, Yuya; Yoshimura, Shige H. [Graduate School of Biostudies, Kyoto University, Kyoto 606-8501 (Japan); Horigome, Tsuneyoshi [Graduate School of Science and Technology, Niigata University, Niigata 950-2181 (Japan); Takeyasu, Kunio [Graduate School of Biostudies, Kyoto University, Kyoto 606-8501 (Japan)

    2013-12-10

    To uncover the molecular composition and dynamics of the functional scaffold for the nucleus, three fractions of biochemically-stable nuclear protein complexes were extracted and used as immunogens to produce a variety of monoclonal antibodies. Many helix-based cytoskeletal proteins were identified as antigens, suggesting their dynamic contribution to nuclear architecture and function. Interestingly, sets of antibodies distinguished distinct subcellular localization of a single isoform of certain cytoskeletal proteins; distinct molecular forms of keratin and actinin were found in the nucleus. Their nuclear shuttling properties were verified by the apparent nuclear accumulations under inhibition of CRM1-dependent nuclear export. Nuclear keratins do not take an obvious filamentous structure, as was revealed by non-filamentous cytoplasmic keratin-specific monoclonal antibody. These results suggest the distinct roles of the helix-based cytoskeletal proteins in the nucleus. - Highlights: • A set of monoclonal antibodies were raised against nuclear scaffold proteins. • Helix-based cytoskeletal proteins were involved in nuclear scaffold. • Many cytoskeletal components shuttle into the nucleus in a CRM1-dependent manner. • Sets of antibodies distinguished distinct subcellular localization of a single isoform. • Nuclear keratin is soluble and does not form an obvious filamentous structure.

  11. Analyzing import intermediates of mitochondrial proteins by blue native gel electrophoresis.

    Science.gov (United States)

    Waizenegger, Thomas; Rapaport, Doron

    2007-01-01

    Blue native gel electrophoresis (BNGE) is a powerful tool for analyzing native protein complexes from biological membranes as well as water-soluble proteins. It can be used for determining relative molecular masses of protein complexes and their subunit composition and for the detection of subcomplexes. We describe the analysis by BNGE of in vitro import reactions composed of radiolabeled precursor proteins and isolated mitochondria. Such an analysis is a powerful tool to follow import intermediates and to study assembly of protein complexes. Analysis of import reactions by BNGE provides information on the molecular mass of the complex with which the imported precursor is associated. In addition, components of such a complex can be identified by incubating the mitochondrial lysate with either soluble antibodies or antibodies coupled to protein A matrix. The binding of soluble antibodies to specific complexes results in an observed shift in their apparent molecular mass (antibody shift). Alternatively, addition of matrix-bound antibodies followed by removal of the matrix from the mixture will result in depletion of the specific complex from the mitochondrial lysate (antibody depletion). The experimental details of these techniques are described.

  12. The physical chemistry of the amyloid phenomenon: thermodynamics and kinetics of filamentous protein aggregation.

    Science.gov (United States)

    Buell, Alexander K; Dobson, Christopher M; Knowles, Tuomas P J

    2014-01-01

    In this chapter, we present an overview of the kinetics and thermodynamics of protein aggregation into amyloid fibrils. The perspective we adopt is largely experimental, but we also discuss recent developments in data analysis and we show that only a combination of well-designed experiments with appropriate theoretical modelling is able to provide detailed mechanistic insight into the complex pathways of amyloid formation. In the first part of the chapter, we describe measurements of the thermodynamic stability of the amyloid state with respect to the soluble state of proteins, as well as the magnitude and origin of this stability. In the second part, we discuss in detail the kinetics of the individual molecular steps in the overall mechanism of the conversion of soluble protein into amyloid fibrils. Finally, we highlight the effects of external factors, such as salt type and concentration, chemical denaturants and molecular chaperones on the kinetics of aggregation.

  13. A sandwiched-culture technique for evaluation of heterologous protein production in a filamentous fungus

    NARCIS (Netherlands)

    Asgeirsdottir, SA; Scholtmeijer, K; Wessels, JGH

    Aspergillus niger is known for its efficient excretion machinery. However, problems have often arisen in obtaining high amounts of heterologous proteins in the culture medium. Here we present a quick method using sandwiched colonies to evaluate transgenic strains for secretion of heterologous

  14. Pseudomonas fluorescens filamentous hemagglutinin, an iron-regulated protein, is an important virulence factor that modulates bacterial pathogenicity

    Directory of Open Access Journals (Sweden)

    Yuan-yuan Sun

    2016-08-01

    Full Text Available Pseudomonas fluorescens is a common bacterial pathogen to a wide range of aquaculture animals including various species of fish. In this study, we employed proteomic analysis and identified filamentous hemagglutinin (FHA as an iron-responsive protein secreted by TSS, a pathogenic P. fluorescens isolate. In vitro study showed that compared to the wild type, the fha mutant TSSfha (i exhibited a largely similar vegetative growth profile but significantly retarded in the ability of biofilm growth and producing extracellular matrix, (ii displayed no apparent flagella and motility, (iii was defective in the attachment to host cells and unable to form self-aggregation, (iv displayed markedly reduced capacity of hemagglutination and surviving in host serum. In vivo infection analysis revealed that TSSfha was significantly attenuated in the ability of dissemination in fish tissues and inducing host mortality, and that antibody blocking of the natural FHA produced by the wild type TSS impaired the infectivity of the pathogen. Furthermore, when introduced into turbot as a subunit vaccine, recombinant FHA elicited a significant protection against lethal TSS challenge. Taken together, these results indicate for the first time that P. fluorescens FHA is a key virulence factor essential to multiple biological processes associated with pathogenicity.

  15. A novel glycan modifies the flagellar filament proteins of the oral bacterium Treponema denticola.

    Science.gov (United States)

    Kurniyati, Kurni; Kelly, John F; Vinogradov, Evgeny; Robotham, Anna; Tu, Youbing; Wang, Juyu; Liu, Jun; Logan, Susan M; Li, Chunhao

    2017-01-01

    While protein glycosylation has been reported in several spirochetes including the syphilis bacterium Treponema pallidum and Lyme disease pathogen Borrelia burgdorferi, the pertinent glycan structures and their roles remain uncharacterized. Herein, a novel glycan with an unusual chemical composition and structure in the oral spirochete Treponema denticola, a keystone pathogen of periodontitis was reported. The identified glycan of mass 450.2 Da is composed of a monoacetylated nonulosonic acid (Non) with a novel extended N7 acyl modification, a 2-methoxy-4,5,6-trihydroxy-hexanoyl residue in which the Non has a pseudaminic acid configuration (L-glycero-L-manno) and is β-linked to serine or threonine residues. This novel glycan modifies the flagellin proteins (FlaBs) of T. denticola by O-linkage at multiple sites near the D1 domain, a highly conserved region of bacterial flagellins that interact with Toll-like receptor 5. Furthermore, mutagenesis studies demonstrate that the glycosylation plays an essential role in the flagellar assembly and motility of T. denticola. To our knowledge, this novel glycan and its unique modification sites have not been reported previously in any bacteria. © 2016 John Wiley & Sons Ltd.

  16. New immunoassay for the mapping of neurofibrillary degeneration in Alzheimer's disease using two monoclonal antibodies against human paired helical filament tau proteins.

    Science.gov (United States)

    Condamines, O; Buée-Scherrer, V; Boissier, L; Wattez, A; Delacourte, A; Pau, B; Mourton-Gilles, C

    1995-06-09

    Monoclonal antibodies against human paired helical filament tau (PHF-tau) proteins were produced. Two of these antibodies, AD1 and AD2, were shown by immunoblot to be directed against distinct hyperphosphorylated epitopes of the PHF-tau proteins. Using AD1 and AD2, an antigen-capture ELISA specific for PHF-tau proteins was developed and used to map the neurofibrillary degeneration of several Broadmann areas from an Alzheimer's disease patient. The results confirm that the neurofibrillary degeneration predominates in parietal and temporal lobes.

  17. Crystal structure of the left-handed archaeal RadA helical filament: identification of a functional motif for controlling quaternary structures and enzymatic functions of RecA family proteins

    Science.gov (United States)

    Chen, Li-Tzu; Ko, Tzu-Ping; Chang, Yuan-Chih; Lin, Kuei-An; Chang, Chia-Seng; Wang, Andrew H.-J.; Wang, Ting-Fang

    2007-01-01

    The RecA family of proteins mediates homologous recombination, an evolutionarily conserved pathway that maintains genomic stability by protecting against DNA double strand breaks. RecA proteins are thought to facilitate DNA strand exchange reactions as closed-rings or as right-handed helical filaments. Here, we report the crystal structure of a left-handed Sulfolobus solfataricus RadA helical filament. Each protomer in this left-handed filament is linked to its neighbour via interactions of a β-strand polymerization motif with the neighbouring ATPase domain. Immediately following the polymerization motif, we identified an evolutionarily conserved hinge region (a subunit rotation motif) in which a 360° clockwise axial rotation accompanies stepwise structural transitions from a closed ring to the AMP–PNP right-handed filament, then to an overwound right-handed filament and finally to the left-handed filament. Additional structural and functional analyses of wild-type and mutant proteins confirmed that the subunit rotation motif is crucial for enzymatic functions of RecA family proteins. These observations support the hypothesis that RecA family protein filaments may function as rotary motors. PMID:17329376

  18. Intermediate Reprogramming of Mouse Skin Fibroblasts into Stem-Like Cells by Bone Morphogenetic Protein 4.

    Science.gov (United States)

    Lee, Seung-Eun; Uhm, Sang-Jun; Son, Yeo-Jin; Park, Yun-Gwi; Kim, Eun-Young; Park, Se-Pill

    2017-04-01

    Specific transcription factors are sufficient to reprogram fully induced pluripotent stem cells or other types of cells. These findings raise the question of whether chemical molecules or proteins can replace transcription factors to alter the defined cell fate. In this study, we treated mouse skin fibroblasts (MSFs) with bone morphogenetic protein 4 (BMP4) and examined intermediate reprogramming of MSFs into stem-like cells. Putative epidermal stem cells isolated from the ventral skin epidermis of an adult mouse were used to confirm the reprogramming activity of BMP4, which increased the proliferation of these cells. After these cells formed spheroids, they were treated with BMP4 and cultured for 5 days. Following BMP4 treatment, the characteristics of these cells changed, and they expressed Oct-4 and its target transcripts Nanog, Sox2, and alkaline phosphatase. To confirm the stem cell potency of these cells, we induced their differentiation into cardiomyocytes. Stem-like cell-derived cardiomyocytes exhibited mRNA expression of cardiac mesoderm markers such as Nk2 transcription factor-related locus 5 and connexin 40, and the cardiomyocyte marker troponin T. These differentiated cells exhibited contracting masses. These results suggest that BMP4-mediated somatic stem cell reprogramming may become an alternative approach for cell therapy.

  19. Association of bacteriochlorophyll a with the CsmA protein in chlorosomes of the photosynthetic green filamentous bacterium Chloroflexus aurantiacus

    DEFF Research Database (Denmark)

    Sakuragi, Y; Frigaard, N-U; Shimada, K

    1999-01-01

    The protein assumed to be associated with bacteriochlorophyll (BChl) a in chlorosomes from the photosynthetic green filamentous bacterium Chloroflexus aurantiacus was investigated by alkaline treatment, proteolytic digestion and a new treatment using 1-hexanol, sodium cholate and Triton X-100. Upon...... alkaline treatment, only the 5.7 kDa CsmA protein was removed from the chlorosomes among six proteins detected by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) analysis, concomitantly with the disappearance of BChl a absorption at 795 nm. Trypsin treatment removed two proteins...... of BChl a to CsmA in isolated chlorosomes, which was estimated to be between 1.2 and 2.7 by amino acid analysis of the SDS-PAGE-resolved protein bands....

  20. Ca{sup 2+}/calmodulin-dependent protein kinase phosphatase (CaMKP/PPM1F) interacts with neurofilament L and inhibits its filament association

    Energy Technology Data Exchange (ETDEWEB)

    Ozaki, Hana [Laboratory of Molecular Brain Science, Graduate School of Integrated Arts and Sciences, Hiroshima University, Higashi-Hiroshima, 739-8521 (Japan); Katoh, Tsuyoshi [Department of Biochemistry, Asahikawa Medical University, Asahikawa, 078-8510 (Japan); Nakagawa, Ryoko; Ishihara, Yasuhiro [Laboratory of Molecular Brain Science, Graduate School of Integrated Arts and Sciences, Hiroshima University, Higashi-Hiroshima, 739-8521 (Japan); Sueyoshi, Noriyuki; Kameshita, Isamu [Department of Life Sciences, Faculty of Agriculture, Kagawa University, Kagawa, 761-0795 (Japan); Taniguchi, Takanobu [Department of Biochemistry, Asahikawa Medical University, Asahikawa, 078-8510 (Japan); Hirano, Tetsuo; Yamazaki, Takeshi [Laboratory of Molecular Brain Science, Graduate School of Integrated Arts and Sciences, Hiroshima University, Higashi-Hiroshima, 739-8521 (Japan); Ishida, Atsuhiko, E-mail: aishida@hiroshima-u.ac.jp [Laboratory of Molecular Brain Science, Graduate School of Integrated Arts and Sciences, Hiroshima University, Higashi-Hiroshima, 739-8521 (Japan)

    2016-09-02

    Ca{sup 2+}/calmodulin-dependent protein kinase phosphatase (CaMKP/PPM1F) is a Ser/Thr phosphatase that belongs to the PPM family. Growing evidence suggests that PPM phosphatases including CaMKP act as a complex with other proteins to regulate cellular functions. In this study, using the two-dimensional far-western blotting technique with digoxigenin-labeled CaMKP as a probe, in conjunction with peptide mass fingerprinting analysis, we identified neurofilament L (NFL) as a CaMKP-binding protein in a Triton-insoluble fraction of rat brain. We confirmed binding of fluorescein-labeled CaMKP (F-CaMKP) to NFL in solution by fluorescence polarization. The analysis showed that the dissociation constant of F-CaMKP for NFL is 73 ± 17 nM (n = 3). Co-immunoprecipitation assay using a cytosolic fraction of NGF-differentiated PC12 cells showed that endogenous CaMKP and NFL form a complex in cells. Furthermore, the effect of CaMKP on self-assembly of NFL was examined. Electron microscopy revealed that CaMKP markedly prevented NFL from forming large filamentous aggregates, suggesting that CaMKP-binding to NFL inhibits its filament association. These findings may provide new insights into a novel mechanism for regulating network formation of neurofilaments during neuronal differentiation. - Highlights: • NFL was identified as a CaMKP-binding protein in an insoluble fraction of rat brain. • CaMKP bound to NFL in solution with a K{sub d} value of 73 ± 17 nM. • A CaMKP-NFL complex was found in NGF-differentiated PC12 cells. • CaMKP-binding to NFL inhibited its filament association. • CaMKP may regulate network formation of neurofilaments in neurons.

  1. OneG: a computational tool for predicting cryptic intermediates in the unfolding kinetics of proteins under native conditions.

    Directory of Open Access Journals (Sweden)

    Tambi Richa

    Full Text Available Understanding the relationships between conformations of proteins and their stabilities is one key to address the protein folding paradigm. The free energy change (ΔG of unfolding reactions of proteins is measured by traditional denaturation methods and native hydrogen-deuterium (H/D exchange methods. However, the free energy of unfolding (ΔG(U and the free energy of exchange (ΔG(HX of proteins are not in good agreement, though the experimental conditions of both methods are well matching to each other. The anomaly is due to any one or combinations of the following reasons: (i effects of cis-trans proline isomerisation under equilibrium unfolding reactions of proteins (ii inappropriateness in accounting the baselines of melting curves (iii presence of cryptic intermediates, which may elude the melting curve analysis and (iv existence of higher energy metastable states in the H/D exchange reactions of proteins. Herein, we have developed a novel computational tool, OneG, which accounts the discrepancy between ΔG(U and ΔG(HX of proteins by systematically accounting all the four factors mentioned above. The program is fully automated and requires four inputs: three-dimensional structures of proteins, ΔG(U, ΔG(U(* and residue-specific ΔG(HX determined under EX2-exchange conditions in the absence of denaturants. The robustness of the program has been validated using experimental data available for proteins such as cytochrome c and apocytochrome b(562 and the data analyses revealed that cryptic intermediates of the proteins detected by the experimental methods and the cryptic intermediates predicted by the OneG for those proteins were in good agreement. Furthermore, using OneG, we have shown possible existence of cryptic intermediates and metastable states in the unfolding pathways of cardiotoxin III and cobrotoxin, respectively, which are homologous proteins. The unique application of the program to map the unfolding pathways of proteins under

  2. [Artificial Cysteine Bridges on the Surface of Green Fluorescent Protein Affect Hydration of Its Transition and Intermediate States].

    Science.gov (United States)

    Melnik, T N; Nagibina, G S; Surin, A K; Glukhova, K A; Melnik, B S

    2018-01-01

    Studying the effect of cysteine bridges on different energy levels of multistage folding proteins will enable a better understanding of the process of folding and functioning of globular proteins. In particular, it will create prospects for directed change in the stability and rate of protein folding. In this work, using the method of differential scanning microcalorimetry, we have studied the effect of three cysteine bridges introduced in different structural elements of the green fluorescent protein on the denaturation enthalpies, activation energies, and heat-capacity increments when this protein passes from native to intermediate and transition states. The studies have allowed us to confirm that, with this protein denaturation, the process hardly damages the structure initially, but then changes occur in the protein structure in the region of 4-6 beta sheets. The cysteine bridge introduced in this region decreases the hydration of the second transition state and increases the hydration of the second intermediate state during the thermal denaturation of the green fluorescent protein.

  3. The ultrastructural localization of keratin proteins and carcinoembryonic antigen in malignant mesotheliomas.

    Science.gov (United States)

    Warhol, M J

    1984-09-01

    The immunoultrastructural localization of keratin proteins and carcinoembryonic antigen (CEA) in mesothelioma cells was accomplished with a low-temperature embedding colloidal gold technique. The keratin antiserum labeled only intermediate filaments. These filaments surrounded the cytoplasmic organelles and were inserted into desmosomes. The CEA antiserum labeled cytoplasmic vesicles and droplets. No definite labeling of microvilli was observed.

  4. Cartilage Intermediate Layer Protein and Asporin Polymorphisms Are Independent Risk Factors of Lumbar Disc Degeneration in Male Collegiate Athletes

    OpenAIRE

    Min, Seok-Ki; Nakazato, Koichi; Ishigami, Hideaki; Hiranuma, Kenji

    2014-01-01

    Objective: Lumbar disc degeneration (LDDG), recently reported to have strong genetic determinants, is a major cause of discopathy and lower back pain. However, most studies have only evaluated the effects of a single susceptibility polymorphism. Our purpose was to examine the effect of two susceptibility polymorphism for LDDG in Japanese collegiate athletes. Design: We investigated two susceptibility genes for LDDG?cartilage intermediate layer protein (CILP) and asporin (ASPN)?in 516 collegia...

  5. Localization of recombination proteins and Srs2 reveals anti-recombinase function in vivo

    DEFF Research Database (Denmark)

    Burgess, Rebecca C; Lisby, Michael; Altmannova, Veronika

    2009-01-01

    Homologous recombination (HR), although an important DNA repair mechanism, is dangerous to the cell if improperly regulated. The Srs2 "anti-recombinase" restricts HR by disassembling the Rad51 nucleoprotein filament, an intermediate preceding the exchange of homologous DNA strands. Here, we...... removes Rad51 indiscriminately from DNA, while the Rad52 protein coordinates appropriate filament reformation. This constant breakdown and rebuilding of filaments may act as a stringent quality control mechanism during HR....

  6. Chaperonin filaments: The archaeal cytoskeleton?

    Science.gov (United States)

    Trent, Jonathan D.; Kagawa, Hiromi K.; Yaoi, Takuro; Olle, Eric; Zaluzec, Nestor J.

    1997-01-01

    Chaperonins are high molecular mass double-ring structures composed of 60-kDa protein subunits. In the hyperthermophilic archaeon Sulfolobus shibatae the two chaperonin proteins represent ≈4% of its total protein and have a combined intracellular concentration of >30 mg/ml. At concentrations ≥ 0.5 mg/ml purified chaperonins form filaments in the presence of Mg2+ and nucleotides. Filament formation requires nucleotide binding (not hydrolysis), and occurs at physiological temperatures in biologically relevant buffers, including a buffer made from cell extracts. These observations suggest that chaperonin filaments may exist in vivo and the estimated 4600 chaperonins per cell suggest that such filaments could form an extensive cytostructure. We observed filamentous structures in unfixed, uranyl-acetate-stained S. shibatae cells, which resemble the chaperonin filaments in size and appearance. ImmunoGold (Janssen) labeling using chaperonin antibodies indicated that many chaperonins are associated with insoluble cellular structures and these structures appear to be filamentous in some areas, although they could not be uranyl-acetate-stained. The existence of chaperonin filaments in vivo suggests a mechanism whereby their protein-folding activities can be regulated. More generally, the filaments themselves may play a cytoskeletal role in Archaea. PMID:9144246

  7. Production of a defensin-like antifungal protein NFAP from Neosartorya fischeri in Pichia pastoris and its antifungal activity against filamentous fungal isolates from human infections.

    Science.gov (United States)

    Virágh, Máté; Vörös, Dóra; Kele, Zoltán; Kovács, Laura; Fizil, Ádám; Lakatos, Gergely; Maróti, Gergely; Batta, Gyula; Vágvölgyi, Csaba; Galgóczy, László

    2014-02-01

    Neosartorya fischeri NRRL 181 isolate secretes a defensin-like antifungal protein (NFAP) which has a remarkable antifungal effect against ascomycetous filamentous fungi. This protein is a promising antifungal agent of biotechnological value; however in spite of the available knowledge of the nature of its 5'-upstream transcriptional regulation elements, the bulk production of NFAP has not been resolved yet. In this study we carried out its heterologous expression in the yeast Pichia pastoris and investigated the growth inhibition effect exerted by the heterologous NFAP (hNFAP) on filamentous fungal isolates from human infections compared with what was caused by the native NFAP. P. pastoris KM71H transformant strain harboring the pPICZαA plasmid with the mature NFAP encoding gene produced the protein. The final yield of the hNFAP was sixfold compared to the NFAP produced by N. fischeri NRRL 181. Based on the signal dispersion of the amide region, it was proven that the hNFAP exists in folded state. The purified hNFAP effectively inhibited the growth of fungal isolates belonging to the Aspergillus and to the Fusarium genus, but all investigated zygomycetous strain proved to be insusceptible. There was no significant difference between the growth inhibition effect exerted by the native and the heterologous NFAP. These data indicated that P. pastoris KM71H can produce the NFAP in an antifungally active folded state. Our results provide a base for further research, e.g., investigation the connection between the protein structure and the antifungal activity using site directed mutagenesis. Copyright © 2013 Elsevier Inc. All rights reserved.

  8. Early Events, Kinetic Intermediates and the Mechanism of Protein Folding in Cytochrome c

    Directory of Open Access Journals (Sweden)

    David S. Kliger

    2009-04-01

    Full Text Available Kinetic studies of the early events in cytochrome c folding are reviewed with a focus on the evidence for folding intermediates on the submillisecond timescale. Evidence from time-resolved absorption, circular dichroism, magnetic circular dichroism, fluorescence energy and electron transfer, small-angle X-ray scattering and amide hydrogen exchange studies on the t £ 1 ms timescale reveals a picture of cytochrome c folding that starts with the ~ 1-ms conformational diffusion dynamics of the unfolded chains. A fractional population of the unfolded chains collapses on the 1 – 100 ms timescale to a compact intermediate IC containing some native-like secondary structure. Although the existence and nature of IC as a discrete folding intermediate remains controversial, there is extensive high time-resolution kinetic evidence for the rapid formation of IC as a true intermediate, i.e., a metastable state separated from the unfolded state by a discrete free energy barrier. Final folding to the native state takes place on millisecond and longer timescales, depending on the presence of kinetic traps such as heme misligation and proline mis-isomerization. The high folding rates observed in equilibrium molten globule models suggest that IC may be a productive folding intermediate. Whether it is an obligatory step on the pathway to the high free energy barrier associated with millisecond timescale folding to the native state, however, remains to be determined.

  9. Assembly of presynaptic filaments. Factors affecting the assembly of RecA protein onto single-stranded DNA

    DEFF Research Database (Denmark)

    Thresher, RJ; Christiansen, Gunna; Griffith, JD

    1988-01-01

    We have previously shown that the assembly of RecA protein onto single-stranded DNA (ssDNA) facilitated by SSB protein occurs in three steps: (1) rapid binding of SSB protein to the ssDNA; (2) nucleation of RecA protein onto this template; and (3) co-operative polymerization of additional Rec...... assembled onto ssDNA at net rates that varied from 250 to 900 RecA protein monomers per minute, with the rate inversely related to the concentration of SSB protein. Combined sucrose sedimentation and electron microscope analysis established that SSB protein was displaced from the ssDNA during RecA protein...

  10. The Novel Desmin Mutant p.A120D Impairs Filament Formation, Prevents Intercalated Disk Localization, and Causes Sudden Cardiac Death

    DEFF Research Database (Denmark)

    Brodehl, Andreas; Dieding, Mareike; Klauke, Bärbel

    2013-01-01

    The intermediate filament protein desmin is encoded by the gene DES and contributes to the mechanical stabilization of the striated muscle sarcomere and cell contacts within the cardiac intercalated disk. DES mutations cause severe skeletal and cardiac muscle diseases with heterogeneous phenotype...

  11. Boolean gates on actin filaments

    Energy Technology Data Exchange (ETDEWEB)

    Siccardi, Stefano, E-mail: ssiccardi@2ssas.it [The Unconventional Computing Centre, University of the West of England, Bristol (United Kingdom); Tuszynski, Jack A., E-mail: jackt@ualberta.ca [Department of Oncology, University of Alberta, Edmonton, Alberta (Canada); Adamatzky, Andrew, E-mail: andrew.adamatzky@uwe.ac.uk [The Unconventional Computing Centre, University of the West of England, Bristol (United Kingdom)

    2016-01-08

    Actin is a globular protein which forms long polar filaments in the eukaryotic cytoskeleton. Actin networks play a key role in cell mechanics and cell motility. They have also been implicated in information transmission and processing, memory and learning in neuronal cells. The actin filaments have been shown to support propagation of voltage pulses. Here we apply a coupled nonlinear transmission line model of actin filaments to study interactions between voltage pulses. To represent digital information we assign a logical TRUTH value to the presence of a voltage pulse in a given location of the actin filament, and FALSE to the pulse's absence, so that information flows along the filament with pulse transmission. When two pulses, representing Boolean values of input variables, interact, then they can facilitate or inhibit further propagation of each other. We explore this phenomenon to construct Boolean logical gates and a one-bit half-adder with interacting voltage pulses. We discuss implications of these findings on cellular process and technological applications. - Highlights: • We simulate interaction between voltage pulses using on actin filaments. • We use a coupled nonlinear transmission line model. • We design Boolean logical gates via interactions between the voltage pulses. • We construct one-bit half-adder with interacting voltage pulses.

  12. Yeast Ivy1p Is a Putative I-BAR-domain Protein with pH-sensitive Filament Forming Ability in vitro.

    Science.gov (United States)

    Itoh, Yuzuru; Kida, Kazuki; Hanawa-Suetsugu, Kyoko; Suetsugu, Shiro

    2016-01-01

    Bin-Amphiphysin-Rvs161/167 (BAR) domains mold lipid bilayer membranes into tubules, by forming a spiral polymer on the membrane. Most BAR domains are thought to be involved in forming membrane invaginations through their concave membrane binding surfaces, whereas some members have convex membrane binding surfaces, and thereby mold membranes into protrusions. The BAR domains with a convex surface form a subtype called the inverse BAR (I-BAR) domain or IRSp53-MIM-homology domain (IMD). Although the mammalian I-BAR domains have been studied, those from other organisms remain elusive. Here, we found putative I-BAR domains in Fungi and animal-like unicellular organisms. The fungal protein containing the putative I-BAR-domain is known as Ivy1p in yeast, and is reportedly localized in the vacuole. The phylogenetic analysis of the I-BAR domains revealed that the fungal I-BAR-domain containing proteins comprise a distinct group from those containing IRSp53 or MIM. Importantly, Ivy1p formed a polymer with a diameter of approximately 20 nm in vitro, without a lipid membrane. The filaments were formed at neutral pH, but disassembled when pH was reverted to basic. Moreover, Ivy1p and the I-BAR domain expressed in mammalian HeLa cells was localized at a vacuole-like structure as filaments as revealed by super-resolved microscopy. These data indicate the pH-sensitive polymer forming ability and the functional conservation of Ivy1p in eukaryotic cells.

  13. Fine filament NbTi superconductive composite

    International Nuclear Information System (INIS)

    Hong, S.; Grabinsky, G.; Marancik, W.; Pattanayak, D.

    1986-01-01

    The large superconducting magnet for the high energy physics accelerator requires fine filament composite to minimize the field error due to the persistent current in the filaments. New concepts toward the fine filament composite and its cable fabrication are discussed. Two-stage cables of fine wire with intermediate number of filaments were introduced. The first stage was six wires cables around one and in the second stage this was used to produce a Rutherford cable. The advantage of this process is in the ease of billet fabrication since the number of filaments in a single wire is within the range of easy billet fabrication. The disadvantage is in the cable fabrication. One of the major concerns in the fabrication of fine NbTi filaments composite in a copper matrix is the intermetallic compound formation during the extrusion and heat treatment steps. The hard intermetallic particles degrade the uniformity of the filaments and reduce the critical current density. The process of using Nb barrier between the filaments and copper matrix in order to prevent this CuTi intermetallic particle formation is described

  14. Assembly of presynaptic filaments. Factors affecting the assembly of RecA protein onto single-stranded DNA

    DEFF Research Database (Denmark)

    Thresher, RJ; Christiansen, Gunna; Griffith, JD

    1988-01-01

    We have previously shown that the assembly of RecA protein onto single-stranded DNA (ssDNA) facilitated by SSB protein occurs in three steps: (1) rapid binding of SSB protein to the ssDNA; (2) nucleation of RecA protein onto this template; and (3) co-operative polymerization of additional Rec......M in the presence of 12 mM-Mg2+), and relatively low concentrations of SSB protein (1 monomer per 18 nucleotides). Assembly was depressed threefold when SSB protein was added to one monomer per nine nucleotides. These effects appeared to be exerted at the nucleation step. Following nucleation, RecA protein...... assembled onto ssDNA at net rates that varied from 250 to 900 RecA protein monomers per minute, with the rate inversely related to the concentration of SSB protein. Combined sucrose sedimentation and electron microscope analysis established that SSB protein was displaced from the ssDNA during RecA protein...

  15. Adipocyte protein modification by Krebs cycle intermediates and fumarate ester-derived succination.

    Science.gov (United States)

    Manuel, Allison M; Frizzell, Norma

    2013-11-01

    Protein succination, the non-enzymatic modification of cysteine residues by fumarate, is distinguishable from succinylation, an enzymatic reaction forming an amide bond between lysine residues and succinyl-CoA. Treatment of adipocytes with 30 mM glucose significantly increases protein succination with only a small change in succinylation. Protein succination may be significantly increased intracellularly after treatment with fumaric acid esters, however, the ester must be removed by saponification to permit 2SC-antibody detection of the fumarate adduct.

  16. The KIM-family protein-tyrosine phosphatases use distinct reversible oxidation intermediates: Intramolecular or intermolecular disulfide bond formation.

    Science.gov (United States)

    Machado, Luciana E S F; Shen, Tun-Li; Page, Rebecca; Peti, Wolfgang

    2017-05-26

    The kinase interaction motif (KIM) family of protein-tyrosine phosphatases (PTPs) includes hematopoietic protein-tyrosine phosphatase (HePTP), striatal-enriched protein-tyrosine phosphatase (STEP), and protein-tyrosine phosphatase receptor type R (PTPRR). KIM-PTPs bind and dephosphorylate mitogen-activated protein kinases (MAPKs) and thereby critically modulate cell proliferation and differentiation. PTP activity can readily be diminished by reactive oxygen species (ROS), e.g. H 2 O 2 , which oxidize the catalytically indispensable active-site cysteine. This initial oxidation generates an unstable sulfenic acid intermediate that is quickly converted into either a sulfinic/sulfonic acid (catalytically dead and irreversible inactivation) or a stable sulfenamide or disulfide bond intermediate (reversible inactivation). Critically, our understanding of ROS-mediated PTP oxidation is not yet sufficient to predict the molecular responses of PTPs to oxidative stress. However, identifying distinct responses will enable novel routes for PTP-selective drug design, important for managing diseases such as cancer and Alzheimer's disease. Therefore, we performed a detailed biochemical and molecular study of all KIM-PTP family members to determine their H 2 O 2 oxidation profiles and identify their reversible inactivation mechanism(s). We show that despite having nearly identical 3D structures and sequences, each KIM-PTP family member has a unique oxidation profile. Furthermore, we also show that whereas STEP and PTPRR stabilize their reversibly oxidized state by forming an intramolecular disulfide bond, HePTP uses an unexpected mechanism, namely, formation of a reversible intermolecular disulfide bond. In summary, despite being closely related, KIM-PTPs significantly differ in oxidation profiles. These findings highlight that oxidation protection is critical when analyzing PTPs, for example, in drug screening. © 2017 by The American Society for Biochemistry and Molecular Biology

  17. Effective intermediate-spin iron in O2-transporting heme proteins

    OpenAIRE

    Schuth, Nils; Mebs, Stefan; Huwald, Dennis; Wrzolek, Pierre; Schwalbe, Matthias; Hemschemeier, Anja; Haumann, Michael

    2017-01-01

    Tuning of the iron-oxygen bonding in hemoproteins is crucial for their versatile O2 chemistry. The nature of the Fe-O2 bond in myoglobin (MB) and hemoglobin (HB) has been debated for a long time. High-resolution X-ray Kβ emission and K-edge absorption spectroscopy on MB and HB solutions and quantum chemical (time-dependent) density functional theory and complete-active-space self-consistent-field calculations revealed an essentially ferrous, intermediate-spin iron in an ozone-like configurati...

  18. Nucleic acid binding properties and intermediates of HCV core protein multimerization in Pichia pastoris

    International Nuclear Information System (INIS)

    Acosta-Rivero, Nelson; Rodriguez, Armando; Musacchio, Alexis; Falcon, Viviana; Suarez, Viana M.; Chavez, Liudmila; Morales-Grillo, Juan; Duenas-Carrera, Santiago

    2004-01-01

    Little is known about the in vivo assembly pathway or structure of the hepatitis C virus nucleocapsid. In this work the intermediates of HCcAg multimerization in Pichia pastoris cells and the nucleic acid binding properties of structured nucleocapsid-like particles (NLPs) were studied. Extensive cross-linking was observed for HCcAg after glutaraldehyde treatment. Data suggest that HCcAg exists in dimeric forms probably representing P21-P21, P21-P23, and P23-P23 dimers. In addition, the presence of HCcAg species that might represent trimers and multimers was observed. After sucrose equilibrium density gradient purification and nuclease digestion, NLPs were shown to contain both RNA and DNA molecules. Finally, the analysis by electron microscopy indicated that native NLPs were resistant to nuclease treatment. These results indicated that HCcAg assembles through dimers, trimers, and multimers' intermediates into capsids in P. pastoris cells. Assembly of NLPs in its natural environment might confer stability to these particles by adopting a compact structure

  19. Time-resolved solution X-ray scattering of tobacco mosaic virus coat protein: kinetics and structure of intermediates

    Energy Technology Data Exchange (ETDEWEB)

    Potschka, M.; Kock, M.H.J.; Adams, M.L.; Schuster, T.M.

    1988-11-01

    The kinetics of assembly and disassembly of tobacco mosaic virus coat protein (TMVP) following temperature jumps have been studied by small-angle X-ray scattering and turbidimetry. The structures of the principal aggregates of TMVP oligomers (A protein), intermediate size (helix I) and large size helical rods (helix II), have been characterized by their average radii of gyration of thickness, cross section, and shape obtained from the corresponding regimes of the small-angle scattering pattern. This structural information was obtained within seconds after the temperature-induced initiation of either polymerization or depolymerization and allowed the authors to detect transient intermediates. This methodology made it possible to observe and characterize the structure of a principal intermediate. Taken together with other kinetic information, these data suggest that polymerization of TMVP under virus self-assembly conditions may proceed via a single-layered helical nucleus that contains about 20 subunits. Previous studies have shown that overshoot polymerization of TMVP can occur and result in metastable long helical viruslike rods which subsequently depolymerize and then form short helical rods, depending on the conditions of the final equilibrium state. The longer rods (helix II) are overshoot polymers which form within seconds and contain 17 1/3 subunits per turn (helix IIB), in contrast to the subunit packing arrangement of 16 1/3 subunits per turn found in the shorter helical rods (helix IA). The latter packing arrangement is the one found in TMV. An overall polymerization scheme is proposed for the formation of these two helical forms of TMVP.

  20. S-(2-Succinyl)cysteine: a novel chemical modification of tissue proteins by a Krebs cycle intermediate.

    Science.gov (United States)

    Alderson, Nathan L; Wang, Yuping; Blatnik, Matthew; Frizzell, Norma; Walla, Michael D; Lyons, Timothy J; Alt, Nadja; Carson, James A; Nagai, Ryoji; Thorpe, Suzanne R; Baynes, John W

    2006-06-01

    S-(2-Succinyl)cysteine (2SC) has been identified as a chemical modification in plasma proteins, in the non-mercaptalbumin fraction of human plasma albumin, in human skin collagen, and in rat skeletal muscle proteins and urine. 2SC increases in human skin collagen with age and is increased in muscle protein of diabetic vs. control rats. The concentration of 2SC in skin collagen and muscle protein correlated strongly with that of the advanced glycation/lipoxidation end-product (AGE/ALE), N(epsilon)-(carboxymethyl)lysine (CML). 2SC is formed by a Michael addition reaction of cysteine sulfhydryl groups with fumarate at physiological pH. Fumarate, but not succinate, inactivates the sulfhydryl enzyme, glyceraldehyde-3-phosphate dehydrogenase in vitro, in concert with formation of 2SC. 2SC is the first example of spontaneous chemical modification of protein by a metabolic intermediate in the Krebs cycle. These observations identify fumarate as an endogenous electrophile and suggest a role for fumarate in regulation of metabolism.

  1. Boolean gates on actin filaments

    Science.gov (United States)

    Siccardi, Stefano; Tuszynski, Jack A.; Adamatzky, Andrew

    2016-01-01

    Actin is a globular protein which forms long polar filaments in the eukaryotic cytoskeleton. Actin networks play a key role in cell mechanics and cell motility. They have also been implicated in information transmission and processing, memory and learning in neuronal cells. The actin filaments have been shown to support propagation of voltage pulses. Here we apply a coupled nonlinear transmission line model of actin filaments to study interactions between voltage pulses. To represent digital information we assign a logical TRUTH value to the presence of a voltage pulse in a given location of the actin filament, and FALSE to the pulse's absence, so that information flows along the filament with pulse transmission. When two pulses, representing Boolean values of input variables, interact, then they can facilitate or inhibit further propagation of each other. We explore this phenomenon to construct Boolean logical gates and a one-bit half-adder with interacting voltage pulses. We discuss implications of these findings on cellular process and technological applications.

  2. [Chitinolytic activity of filamentous fungi].

    Science.gov (United States)

    Shubakov, A A; Kucheriavykh, P S

    2004-01-01

    The chitinolytic activity of nine species of filamentous fungi, classified with seven genera (specifically, Aspergillus, Penicillium, Trichoderma, Paecilomyces, Sporotrichum, Beaueria, and Mucor), was studied. When cultured in liquid medium containing 1% crystalline chitin, all fungi produced extracellular chitosans with activity varying from 0.2 U/mg protein (Sporotrichum olivaceum, Mucor sp., etc.) to 4.0-4.2 U/mg protein (Trichoderma lignorum, Aspergillus niger).

  3. The Cobalamin-binding Protein in Zebrafish is an Intermediate Between the Three Cobalamin-binding Proteins in Human

    DEFF Research Database (Denmark)

    Greibe, Eva Holm; Fedosov, Sergey; Nexø, Ebba

    2012-01-01

    In humans, three soluble extracellular cobalamin-binding proteins; transcobalamin (TC), intrinsic factor (IF), and haptocorrin (HC), are involved in the uptake and transport of cobalamin. In this study, we investigate a cobalamin-binding protein from zebrafish (Danio rerio) and summarize current...

  4. Mapping and Identification of Antifungal Peptides in the Putative Antifungal Protein AfpB from the Filamentous Fungus Penicillium digitatum

    OpenAIRE

    Garrigues, Sandra; Gandía Gómez, Mónica; Borics, Attila; Marx, Florentine; Manzanares, Paloma; Marcos López, José Francisco

    2017-01-01

    Antifungal proteins (AFPs) from Ascomycetes are small cysteine-rich proteins that are abundantly secreted and show antifungal activity against non-producer fungi. A gene coding for a class B AFP (AfpB) was previously identified in the genome of the plant pathogen Penicillium digitatum. However, previous attempts to detect the AfpB protein were not successful despite the high expression of the corresponding afpB gene. In this work, the structure of the putative AfpB was modeled. Based on this ...

  5. Structure and dynamics of the membrane-bound form of the filamentous bacteriophage coat proteins by NMR spectroscopy

    International Nuclear Information System (INIS)

    Bogusky, M.J.

    1987-01-01

    The structure and dynamics of the Pf1 and fd bacteriophage coat proteins in detergent micelles are characterized in solution by nuclear magnetic resonance spectroscopy. The coat proteins are found to exist within the bacterial inner cell membrane during viral infection and assembly. The coat proteins serve as a model system to investigate integral membrane proteins as well as the viral infection and assembly processes. The coat protein is insoluble in aqueous or organic solvents and can only be effectively solubilized in the presence of detergents that form micelles or phospholipids that form vesicles. The effective molecular weight of the detergent-micelle complex is ca. 30K daltons. Sequential assignment strategies were ineffective due to short T/sub 2s/ and severe resonance degeneracy. The backbone resonance assignments were completed by the combination of several homo- and heteronuclear correlation techniques with biosynthetic 15 N labelling. 2D NOE experiments were used to locate and characterize the secondary structure of the membrane bound form of the proteins showing them to be largely helical with the hydrophobic core existing in a very stable helix

  6. Stable intermediates determine proteins' primary unfolding sites in the presence of surfactants

    DEFF Research Database (Denmark)

    Petersen, Steen Vang; Andersen, Kell kleiner; Enghild, Jan J.

    2009-01-01

    Despite detailed knowledge of the overall structural changes and stoichiometries of surfactant binding, little is known about which protein regions constitute the preferred sites of attack for initial unfolding. Here we have exposed three proteins to limited proteolysis at anionic (SDS) and catio......Despite detailed knowledge of the overall structural changes and stoichiometries of surfactant binding, little is known about which protein regions constitute the preferred sites of attack for initial unfolding. Here we have exposed three proteins to limited proteolysis at anionic (SDS......) and cationic (DTAC) surfactant concentrations corresponding to specific conformational transitions, using the surfactant-robust broad-specificity proteases Savinase and Alcalase. Cleavage sites are identified by SDS-PAGE and N-terminal sequencing. We observe well-defined cleavage fragments, which suggest......, cleavage sites can be rationalized from the structure of the protein's folding transition state and the position of loops in the native state. Nevertheless, they are more sensitive to choice of surfactant and protease, probably reflecting a heterogeneous and fluctuating ensemble of partially unfolded...

  7. Autophagy-Associated Protein SmATG12 Is Required for Fruiting-Body Formation in the Filamentous Ascomycete Sordaria macrospora.

    Directory of Open Access Journals (Sweden)

    Antonia Werner

    Full Text Available In filamentous fungi, autophagy functions as a catabolic mechanism to overcome starvation and to control diverse developmental processes under normal nutritional conditions. Autophagy involves the formation of double-membrane vesicles, termed autophagosomes that engulf cellular components and bring about their degradation via fusion with vacuoles. Two ubiquitin-like (UBL conjugation systems are essential for the expansion of the autophagosomal membrane: the UBL protein ATG8 is conjugated to the lipid phosphatidylethanolamine and the UBL protein ATG12 is coupled to ATG5. We recently showed that in the homothallic ascomycete Sordaria macrospora autophagy-related genes encoding components of the conjugation systems are required for fruiting-body development and/or are essential for viability. In the present work, we cloned and characterized the S. macrospora (Smatg12 gene. Two-hybrid analysis revealed that SmATG12 can interact with SmATG7 and SmATG3. To examine its role in S. macrospora, we replaced the open reading frame of Smatg12 with a hygromycin resistance cassette and generated a homokaryotic ΔSmatg12 knockout strain, which displayed slower vegetative growth under nutrient starvation conditions and was unable to form fruiting bodies. In the hyphae of S. macrospora EGFP-labeled SmATG12 was detected in the cytoplasm and as punctate structures presumed to be phagophores or phagophore assembly sites. Delivery of EGFP-labelled SmATG8 to the vacuole was entirely dependent on SmATG12.

  8. Cell-biological aspects of the prion protein in transgenic Xenopus intermediate pituitary cells

    NARCIS (Netherlands)

    Rosmalen, J.W.G. van

    2007-01-01

    In mammals, prions are the causative agents of various neurodegenerative diseases (e.g. scrapie, mad cow and Creutzfeldt-Jakob disease) in which the three-dimensional structure of the normal cellular form of the prion protein (PrPC) is misfolded into the infectious scrapie form (PrPSc or prion). In

  9. Ultrastructural localization of keratin proteins in human skin using low-temperature embedding and the protein A-gold technique.

    Science.gov (United States)

    Warhol, M J; Lucocq, J M; Carlemalm, E; Roth, J

    1985-01-01

    Human skin was embedded in Lowicryl K4M and keratin proteins were localized by incubation with antikeratin antisera, followed by protein A-gold. The antikeratin antisera labeled all intermediate filament (tonofilament) structures in all layers of the epidermis. The association of keratin filaments with hemidesmosomes, desmosomes, and keratohyaline granules was clearly visualized. Desmosomes and keratohyaline granules were not labeled by the antikeratin antisera. No nonfilamentous structures were labeled. The technique described is suitable for studying the distribution of keratin filaments in normal and diseased tissue.

  10. The Interaction of Arp2/3 Complex with Actin: Nucleation, High Affinity Pointed End Capping, and Formation of Branching Networks of Filaments

    Science.gov (United States)

    Mullins, R. Dyche; Heuser, John A.; Pollard, Thomas D.

    1998-05-01

    The Arp2/3 complex is a stable assembly of seven protein subunits including two actin-related proteins (Arp2 and Arp3) and five novel proteins. Previous work showed that this complex binds to the sides of actin filaments and is concentrated at the leading edges of motile cells. Here, we show that Arp2/3 complex purified from Acanthamoeba caps the pointed ends of actin filaments with high affinity. Arp2/3 complex inhibits both monomer addition and dissociation at the pointed ends of actin filaments with apparent nanomolar affinity and increases the critical concentration for polymerization at the pointed end from 0.6 to 1.0 μ M. The high affinity of Arp2/3 complex for pointed ends and its abundance in amoebae suggest that in vivo all actin filament pointed ends are capped by Arp2/3 complex. Arp2/3 complex also nucleates formation of actin filaments that elongate only from their barbed ends. From kinetic analysis, the nucleation mechanism appears to involve stabilization of polymerization intermediates (probably actin dimers). In electron micrographs of quick-frozen, deep-etched samples, we see Arp2/3 bound to sides and pointed ends of actin filaments and examples of Arp2/3 complex attaching pointed ends of filaments to sides of other filaments. In these cases, the angle of attachment is a remarkably constant 70 ± 7 degrees. From these in vitro biochemical properties, we propose a model for how Arp2/3 complex controls the assembly of a branching network of actin filaments at the leading edge of motile cells.

  11. Nuclear actin filaments recruit cofilin and actin-related protein 3, and their formation is connected with a mitotic block

    Czech Academy of Sciences Publication Activity Database

    Kalendová, Alžběta; Kalasová, Ilona; Yamazaki, S.; Uličná, Lívia; Harata, M.; Hozák, Pavel

    2014-01-01

    Roč. 142, č. 2 (2014), s. 139-152 ISSN 0948-6143 R&D Projects: GA ČR GAP305/11/2232; GA MŠk LD12063; GA MŠk(CZ) ED1.1.00/02.0109 Institutional support: RVO:68378050 Keywords : nuclear actin * transcription * mitosis * actin-related protein 3 * cofilin Subject RIV: EB - Genetics ; Molecular Biology Impact factor: 2.927, year: 2013

  12. A novel flagellar sheath protein, FcpA, determines filament coiling, translational motility and virulence for the Leptospira spirochete.

    Science.gov (United States)

    Wunder, Elsio A; Figueira, Cláudio P; Benaroudj, Nadia; Hu, Bo; Tong, Brian A; Trajtenberg, Felipe; Liu, Jun; Reis, Mitermayer G; Charon, Nyles W; Buschiazzo, Alejandro; Picardeau, Mathieu; Ko, Albert I

    2016-08-01

    Leptospira are unique among bacteria based on their helical cell morphology with hook-shaped ends and the presence of periplasmic flagella (PF) with pronounced spontaneous supercoiling. The factors that provoke such supercoiling, as well as the role that PF coiling plays in generating the characteristic hook-end cell morphology and motility, have not been elucidated. We have now identified an abundant protein from the pathogen L. interrogans, exposed on the PF surface, and named it Flagellar-coiling protein A (FcpA). The gene encoding FcpA is highly conserved among Leptospira and was not found in other bacteria. fcpA(-) mutants, obtained from clinical isolates or by allelic exchange, had relatively straight, smaller-diameter PF, and were not able to produce translational motility. These mutants lost their ability to cause disease in the standard hamster model of leptospirosis. Complementation of fcpA restored the wild-type morphology, motility and virulence phenotypes. In summary, we identified a novel Leptospira 36-kDa protein, the main component of the spirochete's PF sheath, and a key determinant of the flagella's coiled structure. FcpA is essential for bacterial translational motility and to enable the spirochete to penetrate the host, traverse tissue barriers, disseminate to cause systemic infection and reach target organs. © 2016 John Wiley & Sons Ltd.

  13. Photoreduction and validation of haem-ligand intermediate states in protein crystals byin situsingle-crystal spectroscopy and diffraction.

    Science.gov (United States)

    Kekilli, Demet; Moreno-Chicano, Tadeo; Chaplin, Amanda K; Horrell, Sam; Dworkowski, Florian S N; Worrall, Jonathan A R; Strange, Richard W; Hough, Michael A

    2017-05-01

    Powerful synergies are available from the combination of multiple methods to study proteins in the crystalline form. Spectroscopies which probe the same region of the crystal from which X-ray crystal structures are determined can give insights into redox, ligand and spin states to complement the information gained from the electron-density maps. The correct assignment of crystal structures to the correct protein redox and ligand states is essential to avoid the misinterpretation of structural data. This is a particular concern for haem proteins, which can occupy a wide range of redox states and are exquisitely sensitive to becoming reduced by solvated electrons generated from interactions of X-rays with water molecules in the crystal. Here, single-crystal spectroscopic fingerprinting has been applied to investigate the laser photoreduction of ferric haem in cytochrome c '. Furthermore, in situ X-ray-driven generation of haem intermediates in crystals of the dye-decolourizing-type peroxidase A (DtpA) from Streptomyces lividans is described.

  14. Solar Features - Prominences and Filaments - Filaments

    Data.gov (United States)

    National Oceanic and Atmospheric Administration, Department of Commerce — Filaments are formed in magnetic loops that hold relatively cool, dense gas suspended above the surface of the Sun (David Hathaway/NASA)

  15. Conformational intermediate of the amyloidogenic protein beta 2-microglobulin at neutral pH

    DEFF Research Database (Denmark)

    Heegaard, N H; Sen, J W; Kaarsholm, N C

    2001-01-01

    electrophoresis that two conformers spontaneously exist in aqueous buffers at neutral pH. Upon treatment of wild-type beta(2)-microglobulin with acetonitrile or trifluoroethanol, two conformations were also observed. These conformations were in equilibrium dependent on the sample temperature and the percentage...... of organic solvent present. Circular dichroism showed a loss of beta-structures and gain of alpha-helices. Reversal to the native conformation occurred when removing the organics. Affinity capillary electrophoresis experiments showed increased specific interactions of the nonnative beta(2)-microglobulin...... conformation with the dyes 8-anilino-1-naphthalene sulfonic acid and Congo red. The observations may relate to early folding events prior to amyloid fibrillation and facilitate the development of methods to detect and inhibit pro-amyloid protein and peptide conformations....

  16. C0 and C1 N-terminal Ig domains of myosin binding protein C exert different effects on thin filament activation.

    Science.gov (United States)

    Harris, Samantha P; Belknap, Betty; Van Sciver, Robert E; White, Howard D; Galkin, Vitold E

    2016-02-09

    Mutations in genes encoding myosin, the molecular motor that powers cardiac muscle contraction, and its accessory protein, cardiac myosin binding protein C (cMyBP-C), are the two most common causes of hypertrophic cardiomyopathy (HCM). Recent studies established that the N-terminal domains (NTDs) of cMyBP-C (e.g., C0, C1, M, and C2) can bind to and activate or inhibit the thin filament (TF). However, the molecular mechanism(s) by which NTDs modulate interaction of myosin with the TF remains unknown and the contribution of each individual NTD to TF activation/inhibition is unclear. Here we used an integrated structure-function approach using cryoelectron microscopy, biochemical kinetics, and force measurements to reveal how the first two Ig-like domains of cMyPB-C (C0 and C1) interact with the TF. Results demonstrate that despite being structural homologs, C0 and C1 exhibit different patterns of binding on the surface of F-actin. Importantly, C1 but not C0 binds in a position to activate the TF by shifting tropomyosin (Tm) to the "open" structural state. We further show that C1 directly interacts with Tm and traps Tm in the open position on the surface of F-actin. Both C0 and C1 compete with myosin subfragment 1 for binding to F-actin and effectively inhibit actomyosin interactions when present at high ratios of NTDs to F-actin. Finally, we show that in contracting sarcomeres, the activating effect of C1 is apparent only once low levels of Ca(2+) have been achieved. We suggest that Ca(2+) modulates the interaction of cMyBP-C with the TF in the sarcomere.

  17. Seasonal changes in isoform composition of giant proteins of thick and thin filaments and titin (connectin) phosphorylation level in striated muscles of bears (Ursidae, Mammalia).

    Science.gov (United States)

    Salmov, N N; Vikhlyantsev, I M; Ulanova, A D; Gritsyna, Yu V; Bobylev, A G; Saveljev, A P; Makariushchenko, V V; Maksudov, G Yu; Podlubnaya, Z A

    2015-03-01

    Seasonal changes in the isoform composition of thick and thin filament proteins (titin, myosin heavy chains (MyHCs), nebulin), as well as in the phosphorylation level of titin in striated muscles of brown bear (Ursus arctos) and hibernating Himalayan black bear (Ursus thibetanus ussuricus) were studied. We found that the changes that lead to skeletal muscle atrophy in bears during hibernation are not accompanied by a decrease in the content of nebulin and intact titin-1 (T1) isoforms. However, a decrease (2.1-3.4-fold) in the content of T2 fragments of titin was observed in bear skeletal muscles (m. gastrocnemius, m. longissimus dorsi, m. biceps) during hibernation. The content of the stiffer N2B titin isoform was observed to increase relative to the content of its more compliant N2BA isoform in the left ventricles of hibernating bears. At the same time, in spite of the absence of decrease in the total content of T1 in the myocardium of hibernating brown bear, the content of T2 fragments decreased ~1.6-fold. The level of titin phosphorylation only slightly increased in the cardiac muscle of hibernating brown bear. In the skeletal muscles of brown bear, the level of titin phosphorylation did not vary between seasons. However, changes in the composition of MyHCs aimed at increasing the content of slow (I) and decreasing the content of fast (IIa) isoforms of this protein during hibernation of brown bear were detected. Content of MyHCs I and IIa in the skeletal muscles of hibernating Himalayan black bear corresponded to that in the skeletal muscles of hibernating brown bear.

  18. Ectopic expression of H-1 parvovirus NS1 protein induces alterations in actin filaments and cell death in human normal MRC-5 and transformed MRC-5 SV2 cells.

    Science.gov (United States)

    Wizla, Pierre; Begue, Agnès; Loison, Ingrid; Richard, Audrey; Caillet-Fauquet, Perrine; Stéhelin, Dominique

    2010-05-01

    When grown in human cell lines, oncolytic H-1 parvovirus (H-1PV) replication preferentially occurs in transformed cells, which ultimately die upon infection. H-1PV-induced cytotoxicity is mainly due to P4 promoter-driven NS1 protein expression. Infection of untransformed cells generally does not induce deleterious effects because the P4 promoter is not activated. Here, we show that ectopic CMV-driven NS1 protein expression in normal human MRC-5 cells results in alterations of actin filaments and cell death, and both effects are prevented by a serine 473 mutation. The same substitution preserves actin filaments of transfected MRC-5 SV2 cells, that are MRC-5 transformed counterparts, but does not impair NS1-induced cytotoxicity.

  19. DUAL PARAMETER FLOW-CYTOMETRY FOR DEOXYRIBONUCLEIC-ACID AND INTERMEDIATE FILAMENT PROTEINS OF RESIDUAL MATURE TERATOMA - ALL TUMOR-CELLS ARE ANEUPLOID

    NARCIS (Netherlands)

    LOOIJENGA, LHJ; OOSTERHUIS, JW; RAMAEKERS, FCS; DEJONG, B; BECK, JLM; SLEIJFER, DT; KOOPS, HS; Dam, A.

    Most testicular germ cell tumors of adults are presumably derived from polyploid carcinoma in situ. Thus, one would expect that even highly differentiated teratoma components are aneuploid and that it is unlikely to find diploid tumor cell (sub)populations. We studied 10 residual mature teratomas

  20. Ca2+ improves organization of single-stranded DNA bases in human Rad51 filament, explaining stimulatory effect on gene recombination.

    KAUST Repository

    Fornander, Louise H

    2012-02-22

    Human RAD51 protein (HsRad51) catalyses the DNA strand exchange reaction for homologous recombination. To clarify the molecular mechanism of the reaction in vitro being more effective in the presence of Ca(2+) than of Mg(2+), we have investigated the effect of these ions on the structure of HsRad51 filament complexes with single- and double-stranded DNA, the reaction intermediates. Flow linear dichroism spectroscopy shows that the two ionic conditions induce significantly different structures in the HsRad51/single-stranded DNA complex, while the HsRad51/double-stranded DNA complex does not demonstrate this ionic dependence. In the HsRad51/single-stranded DNA filament, the primary intermediate of the strand exchange reaction, ATP/Ca(2+) induces an ordered conformation of DNA, with preferentially perpendicular orientation of nucleobases relative to the filament axis, while the presence of ATP/Mg(2+), ADP/Mg(2+) or ADP/Ca(2+) does not. A high strand exchange activity is observed for the filament formed with ATP/Ca(2+), whereas the other filaments exhibit lower activity. Molecular modelling suggests that the structural variation is caused by the divalent cation interfering with the L2 loop close to the DNA-binding site. It is proposed that the larger Ca(2+) stabilizes the loop conformation and thereby the protein-DNA interaction. A tight binding of DNA, with bases perpendicularly oriented, could facilitate strand exchange.

  1. Desensitization of myofilaments to Ca2+ as a therapeutic target for hypertrophic cardiomyopathy with mutations in thin filament proteins.

    Science.gov (United States)

    Alves, Marco L; Dias, Fernando A L; Gaffin, Robert D; Simon, Jillian N; Montminy, Eric M; Biesiadecki, Brandon J; Hinken, Aaron C; Warren, Chad M; Utter, Megan S; Davis, Robert T; Sakthivel, Sadayappan; Robbins, Jeffrey; Wieczorek, David F; Solaro, R John; Wolska, Beata M

    2014-04-01

    Hypertrophic cardiomyopathy (HCM) is a common genetic disorder caused mainly by mutations in sarcomeric proteins and is characterized by maladaptive myocardial hypertrophy, diastolic heart failure, increased myofilament Ca(2+) sensitivity, and high susceptibility to sudden death. We tested the following hypothesis: correction of the increased myofilament sensitivity can delay or prevent the development of the HCM phenotype. We used an HCM mouse model with an E180G mutation in α-tropomyosin (Tm180) that demonstrates increased myofilament Ca(2+) sensitivity, severe hypertrophy, and diastolic dysfunction. To test our hypothesis, we reduced myofilament Ca(2+) sensitivity in Tm180 mice by generating a double transgenic mouse line. We crossed Tm180 mice with mice expressing a pseudophosphorylated cardiac troponin I (S23D and S24D; TnI-PP). TnI-PP mice demonstrated a reduced myofilament Ca(2+) sensitivity compared with wild-type mice. The development of pathological hypertrophy did not occur in mice expressing both Tm180 and TnI-PP. Left ventricle performance was improved in double transgenic compared with their Tm180 littermates, which express wild-type cardiac troponin I. Hearts of double transgenic mice demonstrated no changes in expression of phospholamban and sarcoplasmic reticulum Ca(2+) ATPase, increased levels of phospholamban and troponin T phosphorylation, and reduced phosphorylation of TnI compared with Tm180 mice. Moreover, expression of TnI-PP in Tm180 hearts inhibited modifications in the activity of extracellular signal-regulated kinase and zinc finger-containing transcription factor GATA in Tm180 hearts. Our data strongly indicate that reduction of myofilament sensitivity to Ca(2+) and associated correction of abnormal relaxation can delay or prevent development of HCM and should be considered as a therapeutic target for HCM.

  2. Interaction between TATA-Binding Protein (TBP and Multiprotein Bridging Factor-1 (MBF1 from the Filamentous Insect Pathogenic Fungus Beauveria bassiana.

    Directory of Open Access Journals (Sweden)

    Chi Song

    Full Text Available TATA-binding protein (TBP is a ubiquitous component of eukaryotic transcription factors that acts to nucleate assembly and position pre-initiation complexes. Multiprotein bridging factor 1 (MBF1 is thought to interconnect TBP with gene specific transcriptional activators, modulating transcriptional networks in response to specific signal and developmental programs. The insect pathogen, Beauveria bassiana, is a cosmopolitan fungus found in most ecosystems where it acts as an important regulator of insect populations and can form intimate associations with certain plants. In order to gain a better understanding of the function of MBF1 in filamentous fungi, its interaction with TBP was demonstrated. The MBF1 and TBP homologs in B. bassiana were cloned and purified from a heterologous E. coli expression system. Whereas purified BbTBP was shown to be able to bind oligonucleotide sequences containing the TATA-motif (Kd ≈ 1.3 nM including sequences derived from the promoters of the B. bassiana chitinase and protease genes. In contrast, BbMBF1 was unable to bind to these same target sequences. However, the formation of a ternary complex between BbMBF1, BbTBP, and a TATA-containing target DNA sequence was seen in agarose gel electrophoretic mobility shift assays (EMSA. These data indicate that BbMBF1 forms direct interactions with BbTBP, and that the complex is capable of binding to DNA sequences containing TATA-motifs, confirming that BbTBP can link BbMBF1 to target sequences as part of the RNA transcriptional machinery in fungi.

  3. Interaction between TATA-Binding Protein (TBP) and Multiprotein Bridging Factor-1 (MBF1) from the Filamentous Insect Pathogenic Fungus Beauveria bassiana.

    Science.gov (United States)

    Song, Chi; Ortiz-Urquiza, Almudena; Ying, Sheng-Hua; Zhang, Jin-Xia; Keyhani, Nemat O

    2015-01-01

    TATA-binding protein (TBP) is a ubiquitous component of eukaryotic transcription factors that acts to nucleate assembly and position pre-initiation complexes. Multiprotein bridging factor 1 (MBF1) is thought to interconnect TBP with gene specific transcriptional activators, modulating transcriptional networks in response to specific signal and developmental programs. The insect pathogen, Beauveria bassiana, is a cosmopolitan fungus found in most ecosystems where it acts as an important regulator of insect populations and can form intimate associations with certain plants. In order to gain a better understanding of the function of MBF1 in filamentous fungi, its interaction with TBP was demonstrated. The MBF1 and TBP homologs in B. bassiana were cloned and purified from a heterologous E. coli expression system. Whereas purified BbTBP was shown to be able to bind oligonucleotide sequences containing the TATA-motif (Kd ≈ 1.3 nM) including sequences derived from the promoters of the B. bassiana chitinase and protease genes. In contrast, BbMBF1 was unable to bind to these same target sequences. However, the formation of a ternary complex between BbMBF1, BbTBP, and a TATA-containing target DNA sequence was seen in agarose gel electrophoretic mobility shift assays (EMSA). These data indicate that BbMBF1 forms direct interactions with BbTBP, and that the complex is capable of binding to DNA sequences containing TATA-motifs, confirming that BbTBP can link BbMBF1 to target sequences as part of the RNA transcriptional machinery in fungi.

  4. Tungsten Filament Fire

    Science.gov (United States)

    Ruiz, Michael J.; Perkins, James

    2016-01-01

    We safely remove the outer glass bulb from an incandescent lamp and burn up the tungsten filament after the glass is removed. This demonstration dramatically illustrates the necessity of a vacuum or inert gas for the environment surrounding the tungsten filament inside the bulb. Our approach has added historical importance since the incandescent…

  5. Proteomics of Filamentous Fungi

    NARCIS (Netherlands)

    Passel, van M.W.J.; Schaap, P.J.; Graaff, de L.H.

    2013-01-01

    Filamentous fungi, such as Aspergillus niger and Aspergillus oryzae traditionally have had an important role in providing enzymes and enzyme cocktails that are used in food industry. In recent years the genome sequences of many filamentous fungi have become available. This combined with

  6. Cartilage Intermediate Layer Protein and Asporin Polymorphisms Are Independent Risk Factors of Lumbar Disc Degeneration in Male Collegiate Athletes.

    Science.gov (United States)

    Min, Seok-Ki; Nakazato, Koichi; Ishigami, Hideaki; Hiranuma, Kenji

    2014-01-01

    Lumbar disc degeneration (LDDG), recently reported to have strong genetic determinants, is a major cause of discopathy and lower back pain. However, most studies have only evaluated the effects of a single susceptibility polymorphism. Our purpose was to examine the effect of two susceptibility polymorphism for LDDG in Japanese collegiate athletes. We investigated two susceptibility genes for LDDG-cartilage intermediate layer protein (CILP) and asporin (ASPN)-in 516 collegiate athletes and genotyped the risk allele of CILP (1184T/C) and ASPN (D14). LDDG was evaluated using T2-weighted magnetic resonance imaging. By using logistic regression analysis, we found that the ASPN D14 allele and CILP genotype were associated with an increased risk of LDDG in male but not female athletes (CILP CT: odds ratios [OR] = 1.77, 95% confidence interval [CI] = 1.07-2.93; CILP CC: OR = 4.38, 95% CI = 1.42-13.54; ASPN D14: OR = 2.17, 95% CI = 1.10-4.28]. We also found that CILP C and ASPN D14 were independent variables. The ORs with more than two risk alleles were largely increased. The CILP and ASPN polymorphisms are independent genetic risk factors for LDDG in male but not female Japanese collegiate athletes.

  7. Cartilage Intermediate Layer Protein and Asporin Polymorphisms Are Independent Risk Factors of Lumbar Disc Degeneration in Male Collegiate Athletes

    Science.gov (United States)

    Nakazato, Koichi; Ishigami, Hideaki; Hiranuma, Kenji

    2014-01-01

    Objective: Lumbar disc degeneration (LDDG), recently reported to have strong genetic determinants, is a major cause of discopathy and lower back pain. However, most studies have only evaluated the effects of a single susceptibility polymorphism. Our purpose was to examine the effect of two susceptibility polymorphism for LDDG in Japanese collegiate athletes. Design: We investigated two susceptibility genes for LDDG—cartilage intermediate layer protein (CILP) and asporin (ASPN)—in 516 collegiate athletes and genotyped the risk allele of CILP (1184T/C) and ASPN (D14). LDDG was evaluated using T2-weighted magnetic resonance imaging. Results: By using logistic regression analysis, we found that the ASPN D14 allele and CILP genotype were associated with an increased risk of LDDG in male but not female athletes (CILP CT: odds ratios [OR] = 1.77, 95% confidence interval [CI] = 1.07-2.93; CILP CC: OR = 4.38, 95% CI = 1.42-13.54; ASPN D14: OR = 2.17, 95% CI = 1.10-4.28]. We also found that CILP C and ASPN D14 were independent variables. The ORs with more than two risk alleles were largely increased. Conclusions: The CILP and ASPN polymorphisms are independent genetic risk factors for LDDG in male but not female Japanese collegiate athletes. PMID:26069683

  8. Heterologous gene expression in filamentous fungi.

    Science.gov (United States)

    Su, Xiaoyun; Schmitz, George; Zhang, Meiling; Mackie, Roderick I; Cann, Isaac K O

    2012-01-01

    Filamentous fungi are critical to production of many commercial enzymes and organic compounds. Fungal-based systems have several advantages over bacterial-based systems for protein production because high-level secretion of enzymes is a common trait of their decomposer lifestyle. Furthermore, in the large-scale production of recombinant proteins of eukaryotic origin, the filamentous fungi become the vehicle of choice due to critical processes shared in gene expression with other eukaryotic organisms. The complexity and relative dearth of understanding of the physiology of filamentous fungi, compared to bacteria, have hindered rapid development of these organisms as highly efficient factories for the production of heterologous proteins. In this review, we highlight several of the known benefits and challenges in using filamentous fungi (particularly Aspergillus spp., Trichoderma reesei, and Neurospora crassa) for the production of proteins, especially heterologous, nonfungal enzymes. We review various techniques commonly employed in recombinant protein production in the filamentous fungi, including transformation methods, selection of gene regulatory elements such as promoters, protein secretion factors such as the signal peptide, and optimization of coding sequence. We provide insights into current models of host genomic defenses such as repeat-induced point mutation and quelling. Furthermore, we examine the regulatory effects of transcript sequences, including introns and untranslated regions, pre-mRNA (messenger RNA) processing, transcript transport, and mRNA stability. We anticipate that this review will become a resource for researchers who aim at advancing the use of these fascinating organisms as protein production factories, for both academic and industrial purposes, and also for scientists with general interest in the biology of the filamentous fungi. Copyright © 2012 Elsevier Inc. All rights reserved.

  9. Evolution of Filament Barbs

    OpenAIRE

    Liu, Rui; Xu, Yan; Wang, Haimin

    2010-01-01

    We present a selected few cases in which the sense of chirality of filament barbs changed within as short as hours. We investigate in detail a quiescent filament on 2003 September 10 and 11. Of its four barbs displaying such changes only one overlay a small polarity inversion line inside the EUV filament channel (EFC). No magnetic elements with magnitude above the noise level were detected at the endpoints of all barbs. In particular, a pair of barbs first approached toward and then departed ...

  10. Femtosecond Laser Filamentation

    CERN Document Server

    Chin, See Leang

    2010-01-01

    Femtosecond Laser Filamentation gives a comprehensive review of the physics of propagation of intense femtosecond laser pulses in optical media (principally air) and the applications and challenges of this new technique. This book presents the modern understanding of the physics of femtosecond laser pulse propagation, including unusual new effects such as the self-transformation of the pulse into a white light laser pulse, intensity clamping, the physics of multiple filamentation and competition, and how filaments’ ability to melt glass leads to wave guide writing. The potential applications of laser filamentation in atmospheric sensing and the generation of other electromagnetic pulses from the UV to the radio frequency are treated, together with possible future challenges in the excitation of super-excited states of molecules. Exciting new phenomena such as filament induced ultrafast birefringence and the excitation of molecular rotational wave packets and their multiple revivals in air (gases) will also ...

  11. Filament Substructures and their Interrelation

    Science.gov (United States)

    Lin, Y.; Martin, S. F.; Engvold, O.

    The main structural components of solar filaments, their spines, barbs, and legs at the extreme ends of the spine, are illustrated from recent high-resolution observations. The thread-like structures appear to be present in filaments everywhere and at all times. They are the fundamental elements of solar filaments. The interrelation of the spines, barbs and legs are discussed. From observations, we present a conceptual model of the magnetic field of a filament. We suggest that only a single physical model is needed to explain filaments in a continuous spectrum represented by active region filaments at one end and quiescent filaments at the other end.

  12. Filament-filament switching can be regulated by separation between filaments together with cargo motor number.

    Directory of Open Access Journals (Sweden)

    Robert P Erickson

    Full Text Available How intracellular transport controls the probability that cargos switch at intersections between filaments is not well understood. In one hypothesis some motors on the cargo attach to one filament while others attach to the intersecting filament, and the ensuing tug-of-war determines which filament is chosen. We investigate this hypothesis using 3D computer simulations, and discover that switching at intersections increases with the number of motors on the cargo, but is not strongly dependent on motor number when the filaments touch. Thus, simply controlling the number of active motors on the cargo cannot account for in vivo observations that found reduced switching with increasing motor number, suggesting additional mechanisms of regulation. We use simulations to show that one possible way to regulate switching is by simultaneously adjusting the separation between planes containing the crossing filaments and the total number of active motors on the cargo. Heretofore, the effect of filament-filament separation on switching has been unexplored. We find that the switching probability decreases with increasing filament separation. This effect is particularly strong for cargos with only a modest number of motors. As the filament separation increases past the maximum head-to-head distance of the motor, individual motors walking along a filament will be unable to reach the intersecting filament. Thus, any switching requires that other motors on the cargo attach to the intersecting filament and haul the cargo along it, while motor(s engaged on the original filament detach. Further, if the filament separation is large enough, the cargo can have difficulty proceeding along the initial filament because the engaged motors can walk underneath the intersecting filament, but the cargo itself cannot fit between the filaments. Thus, the cargo either detaches entirely from the original filament, or must dip to the side of the initial filament and then pass below

  13. Filament-filament switching can be regulated by separation between filaments together with cargo motor number.

    Science.gov (United States)

    Erickson, Robert P; Gross, Steven P; Yu, Clare C

    2013-01-01

    How intracellular transport controls the probability that cargos switch at intersections between filaments is not well understood. In one hypothesis some motors on the cargo attach to one filament while others attach to the intersecting filament, and the ensuing tug-of-war determines which filament is chosen. We investigate this hypothesis using 3D computer simulations, and discover that switching at intersections increases with the number of motors on the cargo, but is not strongly dependent on motor number when the filaments touch. Thus, simply controlling the number of active motors on the cargo cannot account for in vivo observations that found reduced switching with increasing motor number, suggesting additional mechanisms of regulation. We use simulations to show that one possible way to regulate switching is by simultaneously adjusting the separation between planes containing the crossing filaments and the total number of active motors on the cargo. Heretofore, the effect of filament-filament separation on switching has been unexplored. We find that the switching probability decreases with increasing filament separation. This effect is particularly strong for cargos with only a modest number of motors. As the filament separation increases past the maximum head-to-head distance of the motor, individual motors walking along a filament will be unable to reach the intersecting filament. Thus, any switching requires that other motors on the cargo attach to the intersecting filament and haul the cargo along it, while motor(s) engaged on the original filament detach. Further, if the filament separation is large enough, the cargo can have difficulty proceeding along the initial filament because the engaged motors can walk underneath the intersecting filament, but the cargo itself cannot fit between the filaments. Thus, the cargo either detaches entirely from the original filament, or must dip to the side of the initial filament and then pass below the crossing

  14. Tropomyosin - master regulator of actin filament function in the cytoskeleton.

    Science.gov (United States)

    Gunning, Peter W; Hardeman, Edna C; Lappalainen, Pekka; Mulvihill, Daniel P

    2015-08-15

    Tropomyosin (Tpm) isoforms are the master regulators of the functions of individual actin filaments in fungi and metazoans. Tpms are coiled-coil parallel dimers that form a head-to-tail polymer along the length of actin filaments. Yeast only has two Tpm isoforms, whereas mammals have over 40. Each cytoskeletal actin filament contains a homopolymer of Tpm homodimers, resulting in a filament of uniform Tpm composition along its length. Evidence for this 'master regulator' role is based on four core sets of observation. First, spatially and functionally distinct actin filaments contain different Tpm isoforms, and recent data suggest that members of the formin family of actin filament nucleators can specify which Tpm isoform is added to the growing actin filament. Second, Tpms regulate whole-organism physiology in terms of morphogenesis, cell proliferation, vesicle trafficking, biomechanics, glucose metabolism and organ size in an isoform-specific manner. Third, Tpms achieve these functional outputs by regulating the interaction of actin filaments with myosin motors and actin-binding proteins in an isoform-specific manner. Last, the assembly of complex structures, such as stress fibers and podosomes involves the collaboration of multiple types of actin filament specified by their Tpm composition. This allows the cell to specify actin filament function in time and space by simply specifying their Tpm isoform composition. © 2015. Published by The Company of Biologists Ltd.

  15. Evolution of filament barbs.

    Science.gov (United States)

    Liu, R.; Xu, Y.; Wang, H.

    We present a selected few cases in which the sense of chirality of filament barbs changed within periods as short as hours. We investigate in detail a quiescent filament on 2003 September 10 and 11. Of its four barbs displaying such changes, only one overlays a small polarity inversion line inside the EUV filament channel (EFC). No magnetic elements with magnitude above the noise level were detected at the endpoints of all barbs. In particular, a pair of barbs first approached toward, and then departed from, each other in Halpha , with the barb endpoints migrating as far as ˜ 10 arcsec. We conclude that the evolution of the barbs was driven by flux emergence and cancellation of small bipolar units at the EFC border.

  16. Structure of Flexible Filamentous Plant Viruses

    Energy Technology Data Exchange (ETDEWEB)

    Kendall, Amy; McDonald, Michele; Bian, Wen; Bowles, Timothy; Baumgarten, Sarah C.; Shi, Jian; Stewart, Phoebe L.; Bullitt, Esther; Gore, David; Irving, Thomas C.; Havens, Wendy M.; Ghabrial, Said A.; Wall, Joseph S.; Stubbs, Gerald (IIT); (BU-M); (Vanderbilt); (Kentucky); (BNL)

    2008-10-23

    Flexible filamentous viruses make up a large fraction of the known plant viruses, but in comparison with those of other viruses, very little is known about their structures. We have used fiber diffraction, cryo-electron microscopy, and scanning transmission electron microscopy to determine the symmetry of a potyvirus, soybean mosaic virus; to confirm the symmetry of a potexvirus, potato virus X; and to determine the low-resolution structures of both viruses. We conclude that these viruses and, by implication, most or all flexible filamentous plant viruses share a common coat protein fold and helical symmetry, with slightly less than 9 subunits per helical turn.

  17. Filaments in Lupus I

    Science.gov (United States)

    Takahashi, Satoko; Rodon, J.; De Gregorio-Monsalvo, I.; Plunkett, A.

    2017-06-01

    The mechanisms behind the formation of sub-stellar mass sources are key to determine the populations at the low-mass end of the stellar distribution. Here, we present mapping observations toward the Lupus I cloud in C18O(2-1) and 13CO(2-1) obtained with APEX. We have identified a few velocity-coherent filaments. Each contains several substellar mass sources that are also identified in the 1.1mm continuum data (see also SOLA catalogue presentation). We will discuss the velocity structure, fragmentation properties of the identified filaments, and the nature of the detected sources.

  18. Kinetics of filamentous phage assembly

    Science.gov (United States)

    Ploss, Martin; Kuhn, Andreas

    2010-12-01

    Filamentous phages release their progeny particles by a secretory process without lysing the bacterial cell. By this process about 6 viral particles per min are secreted from each cell. We show here that when the major coat protein (gp8) is provided from a plasmid we observe a phage progeny production rate depending on the induction of gp8 by IPTG. We also show that a transfection of Escherichia coli lacking F-pili is observed using a mutant of M13 that carries an ampicillin resistance gene, and phage particles are secreted in the absence of an F-plasmid. Extruding phage was visualized by atomic force microscopy (AFM) and by transmission electron microscopy (TEM) using gold-labeled antibodies to the major coat protein.

  19. Filamentous Growth in Eremothecium Fungi

    DEFF Research Database (Denmark)

    Oskarsson, Therese

    , this thesis deals with some of the aspects of hyphal growth, which is an important virulence factor for pathogenic fungi infecting both humans and plants. Hyphal establishment through continuous polar growth is a complex process, requiring the careful coordination of a large subset of proteins involved...... in polarity establishment and maintenance, cytoskeleton dynamics and intracellular transport. The first part of this thesis addresses the A. gossypii Arf3 small GTPase and its GEF- and GAP regulators; Yel1 and Gts1, which has been implicated in polar growth in a wide range of organisms. We could demonstrate......-regulatory activity of AgGts1, the protein could have additional actin organizing properties. In the second and third part, this thesis addresses the use of A. gossypii and its relative E. cymbalariae as model organisms for filamentous growth. A series of assays analyzed the capability of Eremothecium genus fungi...

  20. Suicide inactivation of cytochrome P-450 by methoxsalen. Evidence for the covalent binding of a reactive intermediate to the protein moiety

    International Nuclear Information System (INIS)

    Labbe, G.; Descatoire, V.; Beaune, P.; Letteron, P.; Larrey, D.; Pessayre, D.

    1989-01-01

    Incubation of rat liver microsomes with [3H]methoxsalen and NADPH resulted in the covalent binding of a methoxsalen intermediate to proteins comigrating with cytochromes P-450 UT-A, PB-B/D, ISF-G and PCN-E. Binding was increased by pretreatments with phenobarbital, beta-naphthoflavone (beta NF) and dexamethasone. Such pretreatments also increased the loss of CO-binding capacity either after administration of methoxsalen, or after incubation of hepatic microsomes with methoxsalen and NADPH. Immunoprecipitation of the methoxsalen metabolite-protein adducts in phenobarbital-induced microsomes was moderate with anti-UT-A antibodies, but marked with anti-PB-B/D and anti-PCN-E antibodies. Immunoprecipitation was observed also with anti-ISF-G (anti-beta NF-B) antibodies in beta NF-induced microsomes. Methoxsalen (0.25 mM) inhibited markedly the benzphetamine demethylase activity of phenobarbital-induced microsomes and the erythromycin demethylase activity of dexamethasone-induced microsomes. Whereas methoxsalen itself did not produce any binding spectrum, in contrast either in vivo administration of methoxsalen or incubation in vitro with methoxsalen and NADPH resulted in a low-to-high spin conversion of cytochrome P-450 as suggested by the appearance of a spectrum analogous to a type I binding spectrum. This low-to-high spin conversion was apparently due to a methoxsalen intermediate (probably, covalently bound to the protein and preventing partial sixth ligation of the iron). We conclude that suicide inactivation of cytochrome P-450 by methoxsalen is related to the covalent binding of a methoxsalen intermediate to the protein moiety of several cytochrome P-450 isoenzymes (including UT-A, PB-B/D, PCN-E as well as ISF-G and/or beta NF-B)

  1. Filamentous Influenza Virus Enters Cells via Macropinocytosis

    Science.gov (United States)

    Rossman, Jeremy S.; Leser, George P.

    2012-01-01

    Influenza virus is pleiomorphic, producing both spherical (100-nm-diameter) and filamentous (100-nm by 20-μm) virions. While the spherical virions are known to enter host cells through exploitation of clathrin-mediated endocytosis, the entry pathway for filamentous virions has not been determined, though the existence of an alternative, non-clathrin-, non-caveolin-mediated entry pathway for influenza virus has been known for many years. In this study, we confirm recent results showing that influenza virus utilizes macropinocytosis as an alternate entry pathway. Furthermore, we find that filamentous influenza viruses use macropinocytosis as the primary entry mechanism. Virions enter cells as intact filaments within macropinosomes and are trafficked to the acidic late-endosomal compartment. Low pH triggers a conformational change in the M2 ion channel protein, altering membrane curvature and leading to a fragmentation of the filamentous virions. This fragmentation may enable more-efficient fusion between the viral and endosomal membranes. PMID:22875971

  2. Soluble components of the flagellar export apparatus, FliI, FliJ, and FliH, do not deliver flagellin, the major filament protein, from the cytosol to the export gate.

    Science.gov (United States)

    Sajó, Ráchel; Liliom, Károly; Muskotál, Adél; Klein, Agnes; Závodszky, Péter; Vonderviszt, Ferenc; Dobó, József

    2014-11-01

    Flagella, the locomotion organelles of bacteria, extend from the cytoplasm to the cell exterior. External flagellar proteins are synthesized in the cytoplasm and exported by the flagellar type III secretion system. Soluble components of the flagellar export apparatus, FliI, FliH, and FliJ, have been implicated to carry late export substrates in complex with their cognate chaperones from the cytoplasm to the export gate. The importance of the soluble components in the delivery of the three minor late substrates FlgK, FlgL (hook-filament junction) and FliD (filament-cap) has been convincingly demonstrated, but their role in the transport of the major filament component flagellin (FliC) is still unclear. We have used continuous ATPase activity measurements and quartz crystal microbalance (QCM) studies to characterize interactions between the soluble export components and flagellin or the FliC:FliS substrate-chaperone complex. As controls, interactions between soluble export component pairs were characterized providing Kd values. FliC or FliC:FliS did not influence the ATPase activity of FliI alone or in complex with FliH and/or FliJ suggesting lack of interaction in solution. Immobilized FliI, FliH, or FliJ did not interact with FliC or FliC:FliS detected by QCM. The lack of interaction in the fluid phase between FliC or FliC:FliS and the soluble export components, in particular with the ATPase FliI, suggests that cells use different mechanisms for the export of late minor substrates, and the major substrate, FliC. It seems that the abundantly produced flagellin does not require the assistance of the soluble export components to efficiently reach the export gate. Copyright © 2014 Elsevier B.V. All rights reserved.

  3. Positrusion Filament Recycling System Project

    Data.gov (United States)

    National Aeronautics and Space Administration — TUI proposes a novel process to produce 3d printer feedstock filament out of scrap ABS on the ISS. Currently the plastic filament materials that most 3d printers use...

  4. Solar Features - Prominences and Filaments

    Data.gov (United States)

    National Oceanic and Atmospheric Administration, Department of Commerce — Prominences and filaments are two manifestations of the same phenomenon. Both prominences and filaments are features formed above the chromosphere by cool dense...

  5. Investigation on the Aggregation Behaviors and Filament Morphology of Tau Protein by a Simple 90° Angle Light-Scattering Assay

    Directory of Open Access Journals (Sweden)

    Hai-Lin Liao

    2013-01-01

    Full Text Available The in vitro aggregation of tau constructs was monitored by a simple 90° angle light-scattering (LS approach which was conducted directly on fluorescence instrument. At the optimum incident wavelength (550 nm, unpolarized, the sensitivity of LS was high enough to detect tau aggregation at micromolar range. The nucleation and elongation, different events in the aggregation process of 4RMBD construct (corresponding with the four repeated units of tau Microtubule Binding Domain could be observed by this approach, as compared with ThS fluorescence assay. The validity of this technique was demonstrated over a range of tau concentrations with different tau filaments. Linear regression of scattering light against concentration yielded the x-intercept, the critical concentrations of tau constructs. The critical concentrations of 4RMBD and its S305N mutant are 5.26 μM and 4.04 μM respectively, indicating point mutation S305N, which is associated with FTDP-17, appear to enhance the heparin-induced tau aggregation in vitro. Furthermore, the slopes of concentration dependence curves, as well as the angle dependence, were discussed based on the filaments morphology examined by electron microscopy and ultrasonication experiment.

  6. Fluid shear stress induction of COX-2 protein and prostaglandin release in cultured MC3T3-E1 osteoblasts does not require intact microfilaments or microtubules.

    Science.gov (United States)

    Norvell, Suzanne M; Ponik, Suzanne M; Bowen, Deidre K; Gerard, Rita; Pavalko, Fredrick M

    2004-03-01

    Cultured osteoblasts express three major types of cytoskeleton: actin microfilaments, microtubules, and intermediate filaments. The cytoskeletal network is thought to play an important role in the transmission and conversion of a mechanical stimulus into a biochemical response. To examine a role for the three different cytoskeletal networks in fluid shear stress-induced signaling in osteoblasts, we individually disrupted actin microfilaments, micro-tubules, and intermediate filaments in MC3T3-E1 osteoblasts with multiple pharmacological agents. We subjected these cells to 90 min of laminar fluid shear stress (10 dyn/cm(2)) and compared the PGE(2) and PGI(2) release and induction of cyclooxygenase-2 protein to control cells with intact cytoskeletons. Disruption of actin microfilaments, microtubules, or intermediate filaments in MC3T3-E1 cells did not prevent a significant fluid shear stress-induced release of PGE(2) or PGI(2). Furthermore, disruption of actin microfilaments or microtubules did not prevent a significant fluid shear stress-induced increase in cyclooxygenase-2 protein levels. Disruption of intermediate filaments with acrylamide did prevent the fluid shear stress-induced increase in cyclooxygenase-2 but also prevented a PGE(2)-induced increase in cyclooxygenase-2. Thus none of the three major cytoskeletal networks are required for fluid shear stress-induced prostaglandin release. Furthermore, although neither actin microfilaments nor microtubules are required for fluid shear stress-induced increase in cyclooxygenase-2 levels, the role of intermediate filaments in regulation of cyclooxygenase-2 expression is less clear.

  7. The gene for a lectin-like protein is transcriptionally activated during sexual development, but is not essential for fruiting body formation in the filamentous fungus Sordaria macrospora

    Directory of Open Access Journals (Sweden)

    Cebula Patricia

    2005-11-01

    Full Text Available Abstract Background The filamentous fungus Sordaria macrospora forms complex three-dimensional fruiting bodies called perithecia that protect the developing ascospores and ensure their proper discharge. In previous microarray analyses, several genes have been identified that are downregulated in sterile mutants compared to the wild type. Among these genes was tap1 (transcript associated with perithecial development, a gene encoding a putative lectin homolog. Results Analysis of tap1 transcript levels in the wild type under conditions allowing only vegetative growth compared to conditions that lead to fruiting body development showed that tap1 is not only downregulated in developmental mutants but is also upregulated in the wild type during fruiting body development. We have cloned and sequenced a 3.2 kb fragment of genomic DNA containing the tap1 open reading frame and adjoining sequences. The genomic region comprising tap1 is syntenic to its homologous region in the closely related filamentous fungus Neurospora crassa. To determine whether tap1 is involved in fruiting body development in S. macrospora, a knockout construct was generated in which the tap1 open reading frame was replaced by the hygromycin B resistance gene hph under the control of fungal regulatory regions. Transformation of the S. macrospora wild type with this construct resulted in a tap1 deletion strain where tap1 had been replaced by the hph cassette. The knockout strain displayed no phenotypic differences under conditions of vegetative growth and sexual development when compared to the wild type. Double mutants carrying the Δtap1 allele in several developmental mutant backgrounds were phenotypically similar to the corresponding developmental mutant strains. Conclusion The tap1 transcript is strongly upregulated during sexual development in S. macrospora; however, analysis of a tap1 knockout strain shows that tap1 is not essential for fruiting body formation in S. macrospora.

  8. Whereas Short-Term Facilitation Is Presynaptic, Intermediate-Term Facilitation Involves Both Presynaptic and Postsynaptic Protein Kinases and Protein Synthesis

    Science.gov (United States)

    Jin, Iksung; Kandel, Eric R.; Hawkins, Robert D.

    2011-01-01

    Whereas short-term plasticity involves covalent modifications that are generally restricted to either presynaptic or postsynaptic structures, long-term plasticity involves the growth of new synapses, which by its nature involves both pre- and postsynaptic alterations. In addition, an intermediate-term stage of plasticity has been identified that…

  9. Filamentous Fungi Fermentation

    DEFF Research Database (Denmark)

    Nørregaard, Anders; Stocks, Stuart; Woodley, John

    2014-01-01

    Filamentous fungi (including microorganisms such as Aspergillus niger and Rhizopus oryzae) represent an enormously important platform for industrial fermentation. Two particularly valuable features are the high yield coefficients and the ability to secrete products. However, the filamentous...... morphology, together with non-Newtonian rheological properties (shear thinning), result in poor oxygen transfer unless sufficient energy is provided to the fermentation. While genomic research may improve the organisms, there is no doubt that to enable further application in future it will be necessary...... to match such research with studies of oxygen transfer and energy supply to high viscosity fluids. Hence, the implementation of innovative solutions (some of which in principle are already possible) will be essential to ensure the further development of such fermentations....

  10. Planck intermediate results: VIII. Filaments between interacting clusters

    DEFF Research Database (Denmark)

    Bartlett, J.G.; Cardoso, J.-F.; Castex, G.

    2013-01-01

    . The Planck satellite has provided hundreds of detections of the hot gas in clusters of galaxies via the thermal Sunyaev-Zel'dovich (tSZ) effect and is an ideal instrument for studying extended low-density media through the tSZ effect. In this paper we use the Planck data to search for signatures...... of this intercluster medium. We obtain a temperature of kT = 7.1 ± 0.9 keV (consistent with previous estimates) and a baryon density of (3.7 ± 0.2) × 10-4 cm -3. Conclusions. The Planck satellite mission has provided the first SZ detection of the hot and diffuse intercluster gas. © 2013 ESO....

  11. Mechanosensing through focal adhesion-anchored intermediate filaments

    Czech Academy of Sciences Publication Activity Database

    Gregor, Martin; Osmanagic-Myers, S.; Burgstaller, G.; Wolfram, M.; Fischer, I.; Walko, G.; Resch, G.P.; Jorgl, A.; Herrmann, H.; Wiche, G.

    2014-01-01

    Roč. 28, č. 2 (2014), s. 715-729 ISSN 0892-6638 R&D Projects: GA MŠk(CZ) ED1.1.00/02.0109 Institutional support: RVO:68378050 Keywords : vimentin * plectin * integrin * activation * cellmotility Subject RIV: EB - Genetics ; Molecular Biology Impact factor: 5.043, year: 2014

  12. The evolution of compositionally and functionally distinct actin filaments.

    Science.gov (United States)

    Gunning, Peter W; Ghoshdastider, Umesh; Whitaker, Shane; Popp, David; Robinson, Robert C

    2015-06-01

    The actin filament is astonishingly well conserved across a diverse set of eukaryotic species. It has essentially remained unchanged in the billion years that separate yeast, Arabidopsis and man. In contrast, bacterial actin-like proteins have diverged to the extreme, and many of them are not readily identified from sequence-based homology searches. Here, we present phylogenetic analyses that point to an evolutionary drive to diversify actin filament composition across kingdoms. Bacteria use a one-filament-one-function system to create distinct filament systems within a single cell. In contrast, eukaryotic actin is a universal force provider in a wide range of processes. In plants, there has been an expansion of the number of closely related actin genes, whereas in fungi and metazoa diversification in tropomyosins has increased the compositional variety in actin filament systems. Both mechanisms dictate the subset of actin-binding proteins that interact with each filament type, leading to specialization in function. In this Hypothesis, we thus propose that different mechanisms were selected in bacteria, plants and metazoa, which achieved actin filament compositional variation leading to the expansion of their functional diversity. © 2015. Published by The Company of Biologists Ltd.

  13. Branched intermediate formation is the slowest step in the protein splicing reaction of the Ala1 KlbA intein from Methanococcus jannaschii.

    Science.gov (United States)

    Saleh, Lana; Southworth, Maurice W; Considine, Nancy; O'Neill, Colleen; Benner, Jack; Bollinger, J Martin; Perler, Francine B

    2011-12-13

    We report the first detailed investigation of the kinetics of protein splicing by the Methanococcus jannaschii KlbA (Mja KlbA) intein. This intein has an N-terminal Ala in place of the nucleophilic Cys or Ser residue that normally initiates splicing but nevertheless splices efficiently in vivo [Southworth, M. W., Benner, J., and Perler, F. B. (2000) EMBO J.19, 5019-5026]. To date, the spontaneous nature of the cis splicing reaction has hindered its examination in vitro. For this reason, we constructed an Mja KlbA intein-mini-extein precursor using intein-mediated protein ligation and engineered a disulfide redox switch that permits initiation of the splicing reaction by the addition of a reducing agent such as dithiothreitol (DTT). A fluorescent tag at the C-terminus of the C-extein permits monitoring of the progress of the reaction. Kinetic analysis of the splicing reaction of the wild-type precursor (with no substitutions in known nucleophiles or assisting groups) at various DTT concentrations shows that formation of the branched intermediate from the precursor is reversible (forward rate constant of 1.5 × 10(-3) s(-1) and reverse rate constant of 1.7 × 10(-5) s(-1) at 42 °C), whereas the productive decay of this intermediate to form the ligated exteins is faster and occurs with a rate constant of 2.2 × 10(-3) s(-1). This finding conflicts with reports about standard inteins, for which Asn cyclization has been assigned as the rate-determining step of the splicing reaction. Despite being the slowest step of the reaction, branched intermediate formation in the Mja KlbA intein is efficient in comparison with those of other intein systems. Interestingly, it also appears that this intermediate is protected against thiolysis by DTT, in contrast to other inteins. Evidence is presented in support of a tight coupling between the N-terminal and C-terminal cleavage steps, despite the fact that the C-terminal single-cleavage reaction occurs in variant Mja KlbA inteins in

  14. Inclusion bodies as potential vehicles for recombinant protein delivery into epithelial cells

    Science.gov (United States)

    2012-01-01

    Background We present the potential of inclusion bodies (IBs) as a protein delivery method for polymeric filamentous proteins. We used as cell factory a strain of E. coli, a conventional host organism, and keratin 14 (K14) as an example of a complex protein. Keratins build the intermediate filament cytoskeleton of all epithelial cells. In order to build filaments, monomeric K14 needs first to dimerize with its binding partner (keratin 5, K5), which is then followed by heterodimer assembly into filaments. Results K14 IBs were electroporated into SW13 cells grown in culture together with a “reporter” plasmid containing EYFP labeled keratin 5 (K5) cDNA. As SW13 cells do not normally express keratins, and keratin filaments are built exclusively of keratin heterodimers (i.e. K5/K14), the short filamentous structures we obtained in this study can only be the result of: a) if both IBs and plasmid DNA are transfected simultaneously into the cell(s); b) once inside the cells, K14 protein is being released from IBs; c) released K14 is functional, able to form heterodimers with EYFP-K5. Conclusions Soluble IBs may be also developed for complex cytoskeletal proteins and used as nanoparticles for their delivery into epithelial cells. PMID:22624805

  15. Filament wound structure and method

    International Nuclear Information System (INIS)

    Dritt, W.S.; Gerth, H.L.; Knight, C.E. Jr.; Pardue, R.M.

    1977-01-01

    A filament wound spherical structure is described comprising a plurality of filament band sets disposed about the surface of a mandrel with each band of each set formed of a continuous filament circumferentially wound about the mandrel a selected number of circuits and with each circuit of filament being wound parallel to and contiguous with an immediate previously wound circuit. Each filament band in each band set is wound at the same helix angle from the axis of revolution of the mandrel and all of the bands of each set are uniformly distributed about the mandrel circumference. The pole-to-equator wall thickness taper associated with each band set, as several contiguous band sets are wound about the mandrel starting at the poles, is accumulative as the band sets are nested to provide a complete filament wound sphere of essentially uniform thickness

  16. Magnetic vortex filament flows

    International Nuclear Information System (INIS)

    Barros, Manuel; Cabrerizo, Jose L.; Fernandez, Manuel; Romero, Alfonso

    2007-01-01

    We exhibit a variational approach to study the magnetic flow associated with a Killing magnetic field in dimension 3. In this context, the solutions of the Lorentz force equation are viewed as Kirchhoff elastic rods and conversely. This provides an amazing connection between two apparently unrelated physical models and, in particular, it ties the classical elastic theory with the Hall effect. Then, these magnetic flows can be regarded as vortex filament flows within the localized induction approximation. The Hasimoto transformation can be used to see the magnetic trajectories as solutions of the cubic nonlinear Schroedinger equation showing the solitonic nature of those

  17. Solar Filament Extraction and Characterizing

    Science.gov (United States)

    Yuan, Yuan; Shih, F. Y.; Jing, J.; Wang, H.

    2010-05-01

    This paper presents a new method to extract and characterize solar filaments from H-alpha full-disk images produced by Big Bear Solar Observatory. A cascading Hough Transform method is designed to identify solar disk center location and radius. Solar disks are segmented from the background, and unbalanced illumination on the surface of solar disks is removed using polynomial surface fitting. And then a localized adaptive thresholding is employed to extract solar filament candidates. After the removal of small solar filament candidates, the remaining larger candidates are used as the seeds of region growing. The procedure of region growing not only connects broken filaments but also generate complete shape for each filament. Mathematical morphology thinning is adopted to produce the skeleton of each filament, and graph theory is used to prune branches and barbs to get the main skeleton. The length and the location of the main skeleton is characterized. The proposed method can help scientists and researches study the evolution of solar filament, for instance, to detect solar filament eruption. The presented method has already been used by Space Weather Research Lab of New Jersey Institute of Technology (http://swrl.njit.edu) to generate the solar filament online catalog using H-alpha full-disk images of Global H-alpha Network (http://swrl.njit.edu/ghn_web/).

  18. Insights into enzyme-substrate interaction and characterization of catalytic al intermediates of a Xylella Fastidiosa antioxidant protein

    International Nuclear Information System (INIS)

    Oliveira, Marcos Antonio de; Cussiol, Jose Renato Rosa; Soares Netto, Luis Eduardo; Guimaraes, Beatriz Gomes; Medrano, Francisco Javier; Gozzo, Fabio Cesar

    2005-01-01

    Plants and animals have developed various strategies to defend themselves from pathogens. One of them is the generation of oxidants such as organic Hydroperoxides (OHP). OHP can be generated through free radicals as well as enzymatic oxidation of unsaturated fatty acids. To counteract this oxidative stress, bacteria have evolved several antioxidant mechanisms. Organic hydroperoxide resistance protein (Ohr) was initially identified as a factor involved in the resistance of bacteria, most of them pathogenic, to OHP, but not H 2 O 2 . We have cloned Ohr gene from Xylella fastidiosa and expressed in in Escherichia coli. The biochemical role of Ohr remained unknown for a long time until this work and the work of Nikolov's group (Cornell University, New York) independently showed that these proteins are thiol dependent peroxidases, whose activity is generated by a reactive cysteine. (author)

  19. Insights into enzyme-substrate interaction and characterization of catalytic al intermediates of a Xylella Fastidiosa antioxidant protein

    Energy Technology Data Exchange (ETDEWEB)

    Oliveira, Marcos Antonio de; Cussiol, Jose Renato Rosa; Soares Netto, Luis Eduardo [Sao Paulo Univ. (USP), SP (Brazil). Inst. de Biociencias. Dept. de Genetica e Biologia Evolutiva; Guimaraes, Beatriz Gomes; Medrano, Francisco Javier; Gozzo, Fabio Cesar [Laboratorio Nacional de Luz Sincrotron (LNLS), Campinas, SP (Brazil). Centro de Biologia Molecular Estrutural

    2005-07-01

    Plants and animals have developed various strategies to defend themselves from pathogens. One of them is the generation of oxidants such as organic Hydroperoxides (OHP). OHP can be generated through free radicals as well as enzymatic oxidation of unsaturated fatty acids. To counteract this oxidative stress, bacteria have evolved several antioxidant mechanisms. Organic hydroperoxide resistance protein (Ohr) was initially identified as a factor involved in the resistance of bacteria, most of them pathogenic, to OHP, but not H{sub 2}O{sub 2}. We have cloned Ohr gene from Xylella fastidiosa and expressed in in Escherichia coli. The biochemical role of Ohr remained unknown for a long time until this work and the work of Nikolov's group (Cornell University, New York) independently showed that these proteins are thiol dependent peroxidases, whose activity is generated by a reactive cysteine. (author)

  20. Actin filament-associated protein 1 (AFAP-1) is a key mediator in inflammatory signaling-induced rapid attenuation of intrinsic P-gp function in human brain capillary endothelial cells.

    Science.gov (United States)

    Hoshi, Yutaro; Uchida, Yasuo; Tachikawa, Masanori; Ohtsuki, Sumio; Terasaki, Tetsuya

    2017-04-01

    The purpose of this study was to identify regulatory molecule(s) involved in the inflammatory signaling-induced decrease in P-glycoprotein (P-gp) efflux function at the blood-brain barrier (BBB) that may occur in brain diseases. We confirmed that in vivo P-gp efflux activity at the BBB was decreased without any change in P-gp protein expression level in a mouse model of acute inflammation induced by 3 mg/kg lipopolysaccharide. In a human BBB model cell line (human brain capillary endothelial cells; hCMEC/D3), 1-h treatment with 10 ng/mL tumor necrosis factor-α (TNF-α; an inflammatory mediator) rapidly reduced P-gp efflux activity, but had no effect on P-gp protein expression level. To clarify the non-transcriptional mechanism that causes the decrease in intrinsic efflux activity of P-gp in acute inflammation, we applied comprehensive quantitative phosphoproteomics to compare hCMEC/D3 cells treated with TNF-α and vehicle (control). Actin filament-associated protein-1 (AFAP-1), MAPK1, and transcription factor AP-1 (AP-1) were significantly phosphorylated in TNF-α-treated cells, and were selected as candidate proteins. In validation experiments, knockdown of AFAP-1 expression blocked the reduction in P-gp efflux activity by TNF-α treatment, whereas inhibition of MAPK function or knockdown of AP-1 expression did not. Quantitative targeted absolute proteomics revealed that the reduction in P-gp activity by TNF-α did not require any change in P-gp protein expression levels in the plasma membrane. Our results demonstrate that AFAP-1 is a key mediator in the inflammatory signaling-induced, translocation-independent rapid attenuation of P-gp efflux activity in human brain capillary endothelial cells. © 2017 International Society for Neurochemistry.

  1. Native-unlike Long-lived Intermediates along the Folding Pathway of the Amyloidogenic Protein β2-Microglobulin Revealed by Real-time Two-dimensional NMR*

    Science.gov (United States)

    Corazza, Alessandra; Rennella, Enrico; Schanda, Paul; Mimmi, Maria Chiara; Cutuil, Thomas; Raimondi, Sara; Giorgetti, Sofia; Fogolari, Federico; Viglino, Paolo; Frydman, Lucio; Gal, Maayan; Bellotti, Vittorio; Brutscher, Bernhard; Esposito, Gennaro

    2010-01-01

    β2-microglobulin (β2m), the light chain of class I major histocompatibility complex, is responsible for the dialysis-related amyloidosis and, in patients undergoing long term dialysis, the full-length and chemically unmodified β2m converts into amyloid fibrils. The protein, belonging to the immunoglobulin superfamily, in common to other members of this family, experiences during its folding a long-lived intermediate associated to the trans-to-cis isomerization of Pro-32 that has been addressed as the precursor of the amyloid fibril formation. In this respect, previous studies on the W60G β2m mutant, showing that the lack of Trp-60 prevents fibril formation in mild aggregating condition, prompted us to reinvestigate the refolding kinetics of wild type and W60G β2m at atomic resolution by real-time NMR. The analysis, conducted at ambient temperature by the band selective flip angle short transient real-time two-dimensional NMR techniques and probing the β2m states every 15 s, revealed a more complex folding energy landscape than previously reported for wild type β2m, involving more than a single intermediate species, and shedding new light into the fibrillogenic pathway. Moreover, a significant difference in the kinetic scheme previously characterized by optical spectroscopic methods was discovered for the W60G β2m mutant. PMID:20028983

  2. Mismatch repair protein hMSH2–hMSH6 recognizes mismatches and forms sliding clamps within a D-loop recombination intermediate

    Science.gov (United States)

    Honda, Masayoshi; Okuno, Yusuke; Hengel, Sarah R.; Martín-López, Juana V.; Cook, Christopher P.; Amunugama, Ravindra; Soukup, Randal J.; Subramanyam, Shyamal; Fishel, Richard; Spies, Maria

    2014-01-01

    High fidelity homologous DNA recombination depends on mismatch repair (MMR), which antagonizes recombination between divergent sequences by rejecting heteroduplex DNA containing excessive nucleotide mismatches. The hMSH2–hMSH6 heterodimer is the first responder in postreplicative MMR and also plays a prominent role in heteroduplex rejection. Whether a similar molecular mechanism underlies its function in these two processes remains enigmatic. We have determined that hMSH2–hMSH6 efficiently recognizes mismatches within a D-loop recombination initiation intermediate. Mismatch recognition by hMSH2–hMSH6 is not abrogated by human replication protein A (HsRPA) bound to the displaced single-stranded DNA (ssDNA) or by HsRAD51. In addition, ATP-bound hMSH2–hMSH6 sliding clamps that are essential for downstream MMR processes are formed and constrained within the heteroduplex region of the D-loop. Moreover, the hMSH2–hMSH6 sliding clamps are stabilized on the D-loop by HsRPA bound to the displaced ssDNA. Our findings reveal similarities and differences in hMSH2–hMSH6 mismatch recognition and sliding-clamp formation between a D-loop recombination intermediate and linear duplex DNA. PMID:24395779

  3. Mycobacterium tuberculosis septum site determining protein, Ssd encoded by rv3660c, promotes filamentation and elicits an alternative metabolic and dormancy stress response

    Directory of Open Access Journals (Sweden)

    Crew Rebecca

    2011-04-01

    Full Text Available Abstract Background Proteins that are involved in regulation of cell division and cell cycle progression remain undefined in Mycobacterium tuberculosis. In addition, there is a growing appreciation that regulation of cell replication at the point of division is important in establishing a non-replicating persistent state. Accordingly, the objective of this study was to use a systematic approach consisting of consensus-modeling bioinformatics, ultrastructural analysis, and transcriptional mapping to identify septum regulatory proteins that participate in adaptive metabolic responses in M. tuberculosis. Results Septum site determining protein (Ssd, encoded by rv3660c was discovered to be an ortholog of septum site regulating proteins in actinobacteria by bioinformatics analysis. Increased expression of ssd in M. smegmatis and M. tuberculosis inhibited septum formation resulting in elongated cells devoid of septa. Transcriptional mapping in M. tuberculosis showed that increased ssd expression elicited a unique response including the dormancy regulon and alternative sigma factors that are thought to play a role in adaptive metabolism. Disruption of rv3660c by transposon insertion negated the unique transcriptional response and led to a reduced bacterial length. Conclusions This study establishes the first connection between a septum regulatory protein and induction of alternative metabolism consisting of alternative sigma factors and the dormancy regulon that is associated with establishing a non-replicating persistent intracellular lifestyle. The identification of a regulatory component involved in cell cycle regulation linked to the dormancy response, whether directly or indirectly, provides a foundation for additional studies and furthers our understanding of the complex mechanisms involved in establishing a non-replicating state and resumption of growth.

  4. Use of the Chinchilla model to evaluate the vaccinogenic potential of the Moraxella catarrhalis filamentous hemagglutinin-like proteins MhaB1 and MhaB2.

    Directory of Open Access Journals (Sweden)

    Teresa L Shaffer

    Full Text Available Moraxella catarrhalis causes significant health problems, including 15-20% of otitis media cases in children and ~10% of respiratory infections in adults with chronic obstructive pulmonary disease. The lack of an efficacious vaccine, the rapid emergence of antibiotic resistance in clinical isolates, and high carriage rates reported in children are cause for concern. In addition, the effectiveness of conjugate vaccines at reducing the incidence of otitis media caused by Streptococcus pneumoniae and nontypeable Haemophilus influenzae suggest that M. catarrhalis infections may become even more prevalent. Hence, M. catarrhalis is an important and emerging cause of infectious disease for which the development of a vaccine is highly desirable. Studying the pathogenesis of M. catarrhalis and the testing of vaccine candidates have both been hindered by the lack of an animal model that mimics human colonization and infection. To address this, we intranasally infected chinchilla with M. catarrhalis to investigate colonization and examine the efficacy of a protein-based vaccine. The data reveal that infected chinchillas produce antibodies against antigens known to be major targets of the immune response in humans, thus establishing immune parallels between chinchillas and humans during M. catarrhalis infection. Our data also demonstrate that a mutant lacking expression of the adherence proteins MhaB1 and MhaB2 is impaired in its ability to colonize the chinchilla nasopharynx, and that immunization with a polypeptide shared by MhaB1 and MhaB2 elicits antibodies interfering with colonization. These findings underscore the importance of adherence proteins in colonization and emphasize the relevance of the chinchilla model to study M. catarrhalis-host interactions.

  5. Comparative Biomechanics of Thick Filaments and Thin Filaments with Functional Consequences for Muscle Contraction

    Directory of Open Access Journals (Sweden)

    Mark S. Miller

    2010-01-01

    Full Text Available The scaffold of striated muscle is predominantly comprised of myosin and actin polymers known as thick filaments and thin filaments, respectively. The roles these filaments play in muscle contraction are well known, but the extent to which variations in filament mechanical properties influence muscle function is not fully understood. Here we review information on the material properties of thick filaments, thin filaments, and their primary constituents; we also discuss ways in which mechanical properties of filaments impact muscle performance.

  6. Aspiperidine oxide, a piperidine N-oxide from the filamentous fungus Aspergillus indologenus

    DEFF Research Database (Denmark)

    Petersen, Lene Maj; Kildgaard, Sara; Jaspars, Marcel

    2015-01-01

    A novel secondary metabolite, aspiperidine oxide, was isolated from the filamentous fungus, Aspergillus indologenus. The structure of aspiperidine oxide was determined from extensive 1D and 2D NMR spectroscopic analysis supported by high-resolution mass spectrometry. The structure revealed a rare...... piperidine N-oxide, not observed in filamentous fungi before. A biosynthetic pathway towards aspiperidine oxide is proposed, based on tentative identification of intermediates from UHPLC-DAD-HRMS data....

  7. Spatial organization of proteins in metastasizing cells.

    Science.gov (United States)

    Rönnlund, Daniel; Gad, Annica K B; Blom, Hans; Aspenström, Pontus; Widengren, Jerker

    2013-09-01

    The ability of tumor cells to invade into the surrounding tissue is linked to defective adhesive and mechanical properties of the cells, which are regulated by cell surface adhesions and the intracellular filamentous cytoskeleton, respectively. With the aim to further reveal the underlying mechanisms and provide new strategies for early cancer diagnostics, we have used ultrahigh resolution stimulated emission depletion (STED) microscopy as a means to identify metastasizing cells, based on their subcellular protein distribution patterns reflecting their specific adhesive and mechanical properties. We have compared the spatial distribution of cell-matrix adhesion sites and the vimentin filamentous systems in a matched pair of primary, normal, and metastatic human fibroblast cells. We found that the metastatic cells showed significantly increased densities and more homogenous distributions of nanoscale adhesion-related particles. Moreover, they showed an increase in the number but reduced sizes of the areas of cell-matrix adhesion complexes. The organization of the vimentin intermediate filaments was also found to be significantly different in the metastasizing cells, showing an increased entanglement and loss of directionality. Image analysis procedures were established, allowing an objective detection and characterization of these features and distinction of metastatic cells from their normal counterparts. In conclusion, our results suggest that STED microscopy provides a novel tool to identify metastasizing cells from a very sparse number of cells, based on the altered spatial distribution of the cell-matrix adhesions and intermediate filaments. Copyright © 2013 International Society for Advancement of Cytometry.

  8. Colloidal transport by active filaments.

    Science.gov (United States)

    Manna, Raj Kumar; Kumar, P B Sunil; Adhikari, R

    2017-01-14

    Enhanced colloidal transport beyond the limit imposed by diffusion is usually achieved through external fields. Here, we demonstrate the ballistic transport of a colloidal sphere using internal sources of energy provided by an attached active filament. The latter is modeled as a chain of chemo-mechanically active beads connected by potentials that enforce semi-flexibility and self-avoidance. The fluid flow produced by the active beads and the forces they mediate are explicitly taken into account in the overdamped equations of motion describing the colloid-filament assembly. The speed and efficiency of transport depend on the dynamical conformational states of the filament. We characterize these states using filament writhe as an order parameter and identify ones yielding maxima in speed and efficiency of transport. The transport mechanism reported here has a remarkable resemblance to the flagellar propulsion of microorganisms which suggests its utility in biomimetic systems.

  9. Colloidal transport by active filaments

    Science.gov (United States)

    Manna, Raj Kumar; Kumar, P. B. Sunil; Adhikari, R.

    2017-01-01

    Enhanced colloidal transport beyond the limit imposed by diffusion is usually achieved through external fields. Here, we demonstrate the ballistic transport of a colloidal sphere using internal sources of energy provided by an attached active filament. The latter is modeled as a chain of chemo-mechanically active beads connected by potentials that enforce semi-flexibility and self-avoidance. The fluid flow produced by the active beads and the forces they mediate are explicitly taken into account in the overdamped equations of motion describing the colloid-filament assembly. The speed and efficiency of transport depend on the dynamical conformational states of the filament. We characterize these states using filament writhe as an order parameter and identify ones yielding maxima in speed and efficiency of transport. The transport mechanism reported here has a remarkable resemblance to the flagellar propulsion of microorganisms which suggests its utility in biomimetic systems.

  10. Real-time Monitoring of Intermediates Reveals the Reaction Pathway in the Thiol-Disulfide Exchange between Disulfide Bond Formation Protein A (DsbA) and B (DsbB) on a Membrane-immobilized Quartz Crystal Microbalance (QCM) System*

    Science.gov (United States)

    Yazawa, Kenjiro; Furusawa, Hiroyuki; Okahata, Yoshio

    2013-01-01

    Disulfide bond formation protein B (DsbBS-S,S-S) is an inner membrane protein in Escherichia coli that has two disulfide bonds (S-S, S-S) that play a role in oxidization of a pair of cysteine residues (SH, SH) in disulfide bond formation protein A (DsbASH,SH). The oxidized DsbAS-S, with one disulfide bond (S-S), can oxidize proteins with SH groups for maturation of a folding preprotein. Here, we have described the transient kinetics of the oxidation reaction between DsbASH,SH and DsbBS-S,S-S. We immobilized DsbBS-S,S-S embedded in lipid bilayers on the surface of a 27-MHz quartz crystal microbalance (QCM) device to detect both formation and degradation of the reaction intermediate (DsbA-DsbB), formed via intermolecular disulfide bonds, as a mass change in real time. The obtained kinetic parameters (intermediate formation, reverse, and oxidation rate constants (kf, kr, and kcat, respectively) indicated that the two pairs of cysteine residues in DsbBS-S,S-S were more important for the stability of the DsbA-DsbB intermediate than ubiquinone, an electron acceptor for DsbBS-S,S-S. Our data suggested that the reaction pathway of almost all DsbASH,SH oxidation processes would proceed through this stable intermediate, avoiding the requirement for ubiquinone. PMID:24145032

  11. Stability of two-dimensional vorticity filaments

    International Nuclear Information System (INIS)

    Elhmaidi, D.; Provenzale, A.; Lili, T.; Babiano, A.

    2004-01-01

    We discuss the results of a numerical study on the stability of two-dimensional vorticity filaments around a circular vortex. We illustrate how the stability of the filaments depends on the balance between the strain associated with the far field of the vortex and the local vorticity of the filament, and we discuss an empirical criterion for filament stability

  12. Urinary Loss of Tricarboxylic Acid Cycle Intermediates As Revealed by Metabolomics Studies: An Underlying Mechanism to Reduce Lipid Accretion by Whey Protein Ingestion?

    Science.gov (United States)

    2015-01-01

    Whey protein intake is associated with the modulation of energy metabolism and altered body composition both in human subjects and in animals, but the underlying mechanisms are not yet elucidated. We fed obesity-prone C57BL/6J mice high-fat diets with either casein (HF casein) or whey (HF whey) for 6 weeks. At equal energy intake and apparent fat and nitrogen digestibility, mice fed HF whey stored less energy as lipids, evident both as lower white adipose tissue mass and as reduced liver lipids, compared with HF-casein-fed mice. Explorative analyses of 48 h urine, both by 1H NMR and LC–MS metabolomic platforms, demonstrated higher urinary excretion of tricarboxylic acid (TCA) cycle intermediates citric acid and succinic acid (identified by both platforms), and cis-aconitic acid and isocitric acid (identified by LC–MS platform) in the HF whey, relative to in the HF-casein-fed mice. Targeted LC–MS analyses revealed higher citric acid and cis-aconitic acid concentrations in fed state plasma, but not in liver of HF-whey-fed mice. We propose that enhanced urinary loss of TCA cycle metabolites drain available substrates for anabolic processes, such as lipogenesis, thereby leading to reduced lipid accretion in HF-whey-fed compared to HF-casein-fed mice. PMID:24702026

  13. Chemical Strategies for the Covalent Modification of Filamentous Phage

    Directory of Open Access Journals (Sweden)

    Matthew B Francis

    2014-12-01

    Full Text Available Historically filamentous bacteriophage have been known to be the workhorse of phage display due to their ability to link genotype to phenotype. More recently, the filamentous phage scaffold has proved to be powerful outside the realms of phage display technology in fields such as molecular imaging, cancer research and materials and vaccine development. The ability of the virion to serve as a platform for a variety of applications heavily relies on the functionalization of the phage coat proteins with a wide variety of functionalities. Genetic modification of the coat proteins has been the most widely used strategy for functionalizing the virion; however complementary chemical modification strategies can help to diversify the range of materials that can be developed. This review emphasizes the recent advances that have been made in the chemical modification of filamentous phage as well as some of the challenges that are involved functionalizing the virion.

  14. Chaperonin filaments : their formation and an evaluation of methods for studying them.

    Energy Technology Data Exchange (ETDEWEB)

    Yaoi, T.; Kagawa, K. H.; Trent, J. D.; Center for Mechanistic Biology and Biotechnology

    1998-08-01

    Chaperonins are multisubunit protein complexes that can be isolated from cells as high-molecular-weight structures that appear as double rings in the electron microscope. We recently discovered that chaperonin double rings isolated from the hyperthermophilic archaeon Sulfolobus shibatae, when incubated at physiological temperatures in the presence of ATP and Mg{sup 2+}, stacked into filaments; we hypothesized that these filaments are related to filaments seen inside S. shibatae cells and that chaperonins exist as filaments in vivo. This paper elucidates the conditions under which we have observed S. shibatae chaperonins to form filaments and evaluates native polyacrylamide gel electrophoresis (PAGE), TEM, spectrophotometry, and centrifugation as methods for studying these filaments. We observed that in the presence of Mg{sup 2+} combined with ATP, ADP, ATP{gamma}S, or GTP, native PAGE indicated that chaperonin subunits assembled into double rings and that the conformation of these double rings was effected by nucleotide binding, but we saw no indication of chaperonin filament formation. Under these same conditions, however, TEM, spectroscopy, and centrifugation methods indicated that chaperonin subunits and double rings had assembled into filaments. We determined that this discrepancy in the representation of the chaperonin structure was due to the native PAGE method itself. When we exposed chaperonin filaments to the electrophoretic field used in native PAGE, the filaments dissociated into double rings. This suggests that TEM, spectrophotometry, and centrifugation are the preferred methods for studying the higher-order structures of chaperonins, which are likely to be of biological significance.

  15. Chaperone role for proteins p618 and p892 in the extracellular tail development of Acidianus two-tailed virus

    DEFF Research Database (Denmark)

    Scheele, Urte; Erdmann, Susanne; Ungewickell, Ernst J.

    2011-01-01

    The crenarchaeal Acidianus two-tailed virus (ATV) undergoes a remarkable morphological development, extracellularly and independently of host cells, by growing long tails at each end of a spindle-shaped virus particle. Initial work suggested that an intermediate filament-like protein, p800...... the interactions observed between the different protein and DNA components and to explain their possible structural and functional roles in extracellular tail development....

  16. Dynamics of RecA filaments on single-stranded DNA

    NARCIS (Netherlands)

    Van Loenhout, M.T.J.; Van der Heijden, T.; Kanaar, R.; Wyman, C.; Dekker, C.

    2009-01-01

    RecA, the key protein in homologous recombination, performs its actions as a helical filament on single-stranded DNA (ssDNA). ATP hydrolysis makes the RecA–ssDNA filament dynamic and is essential for successful recombination. RecA has been studied extensively by single-molecule techniques on

  17. Structural and functional properties of chimeric EspA-FliCi filaments of EPEC.

    Science.gov (United States)

    Crepin, Valerie F; Martinez, Eric; Shaw, Robert K; Frankel, Gad; Daniell, Sarah J

    2008-04-18

    Enteropathogenic Escherichia coli utilise a filamentous type III secretion system to translocate effector proteins into host gut epithelial cells. The primary constituent of the extracellular component of the filamentous type III secretion system is EspA. This forms a long flexible helical conduit between the bacterium and host and has a structure almost identical to that of the flagella filament. We have inserted the D3 domain of FliCi (from Salmonella typhimurium) into the outer domain of EspA and have studied the structure and function of modified filaments when expressed in an enteropathogenic E. coli espA mutant. We found that the chimeric protein EspA-FliCi filaments were biologically active as they supported protein secretion and translocation [assessed by their ability to trigger actin polymerisation beneath adherent bacteria (fluorescent actin staining test)]. The expressed filaments were recognised by both EspA and FliCi antisera. Visualisation and analysis of the chimeric filaments by electron microscopy after negative staining showed that, remarkably, EspA filaments are able to tolerate a large protein insertion without a significant effect on their helical architecture.

  18. Discrete nuclear structures in actively growing neuroblastoma cells are revealed by antibodies raised against phosphorylated neurofilament proteins

    Directory of Open Access Journals (Sweden)

    Raabe Timothy D

    2003-04-01

    Full Text Available Abstract Background Nuclear objects that have in common the property of being recognized by monoclonal antibodies specific for phosphoprotein epitopes and cytoplasmic intermediate filaments (in particular, SMI-31 and RT-97 have been reported in glial and neuronal cells, in situ and in vitro. Since neurofilament and glial filaments are generally considered to be restricted to the cytoplasm, we were interested in exploring the identity of the structures labeled in the nucleus as well as the conditions under which they could be found there. Results Using confocal microscopy and western analysis techniques, we determined 1 the immunolabeled structures are truly within the nucleus; 2 the phosphoepitope labeled by SMI-31 and RT-97 is not specific to neurofilaments (NFs and it can be identified on other intermediate filament proteins (IFs in other cell types; and 3 there is a close relationship between DNA synthesis and the amount of nuclear staining by these antibodies thought to be specific for cytoplasmic proteins. Searches of protein data bases for putative phosphorylation motifs revealed that lamins, NF-H, and GFAP each contain a single tyrosine phosphorylation motif with nearly identical amino acid sequence. Conclusion We therefore suggest that this sequence may be the epitope recognized by SMI-31 and RT-97 mABs, and that the nuclear structures previously reported and shown here are likely phosphorylated lamin intermediate filaments, while the cytoplasmic labeling revealed by the same mABs indicates phosphorylated NFs in neurons or GFAP in glia.

  19. Structural Dynamics of the Vimentin Coiled-coil Contact Regions Involved in Filament Assembly as Revealed by Hydrogen-Deuterium Exchange.

    Science.gov (United States)

    Premchandar, Aiswarya; Mücke, Norbert; Poznański, Jarosław; Wedig, Tatjana; Kaus-Drobek, Magdalena; Herrmann, Harald; Dadlez, Michał

    2016-11-25

    Intermediate filaments (IF) are major constituents of the cytoskeleton of metazoan cells. They are not only responsible for the mechanical properties but also for various physiological activities in different cells and tissues. The building blocks of IFs are extended coiled-coil-forming proteins exhibiting a characteristic central α-helical domain ("rod"). The fundamental principles of the filament assembly mechanism and the network formation have been widely elucidated for the cytoplasmic IF protein vimentin. Also, a comprehensive structural model for the tetrameric complex of vimentin has been obtained by X-ray crystallography in combination with various biochemical and biophysical techniques. To extend these static data and to investigate the dynamic properties of the full-length proteins in solution during the various assembly steps, we analyzed the patterns of hydrogen-deuterium exchange in vimentin and in four variants carrying point mutations in the IF consensus motifs present at either end of the α-helical rod that cause an assembly arrest at the unit-length filament (ULF) stage. The results yielded unique insights into the structural properties of subdomains within the full-length vimentin, in particular in regions of contact in α-helical and linker segments that stabilize different oligomeric forms such as tetramers, ULFs, and mature filaments. Moreover, hydrogen-deuterium exchange analysis of the point-mutated variants directly demonstrated the active role of the IF consensus motifs in the oligomerization mechanism of tetramers during ULF formation. Ultimately, using molecular dynamics simulation procedures, we provide a structural model for the subdomain-mediated tetramer/tetramer interaction via "cross-coiling" as the first step of the assembly process. © 2016 by The American Society for Biochemistry and Molecular Biology, Inc.

  20. Beam distribution function after filamentation

    Energy Technology Data Exchange (ETDEWEB)

    Raubenheimer, T.O.; Decker, F.J.; Seeman, J.T.

    1995-05-01

    In this paper, the authors calculate the beam distribution function after filamentation (phase-mixing) of a focusing mismatch. This distribution is relevant when interpreting beam measurements and sources of emittance dilution in linear colliders. It is also important when considering methods of diluting the phase space density, which may be required for the machine protection system in future linear colliders, and it is important when studying effects of trapped ions which filament in the electron beam potential. Finally, the resulting distribution is compared with measured beam distributions from the SLAC linac.

  1. Simulating the formation of keratin filament networks by a piecewise-deterministic Markov process.

    Science.gov (United States)

    Beil, Michael; Lück, Sebastian; Fleischer, Frank; Portet, Stéphanie; Arendt, Wolfgang; Schmidt, Volker

    2009-02-21

    Keratin intermediate filament networks are part of the cytoskeleton in epithelial cells. They were found to regulate viscoelastic properties and motility of cancer cells. Due to unique biochemical properties of keratin polymers, the knowledge of the mechanisms controlling keratin network formation is incomplete. A combination of deterministic and stochastic modeling techniques can be a valuable source of information since they can describe known mechanisms of network evolution while reflecting the uncertainty with respect to a variety of molecular events. We applied the concept of piecewise-deterministic Markov processes to the modeling of keratin network formation with high spatiotemporal resolution. The deterministic component describes the diffusion-driven evolution of a pool of soluble keratin filament precursors fueling various network formation processes. Instants of network formation events are determined by a stochastic point process on the time axis. A probability distribution controlled by model parameters exercises control over the frequency of different mechanisms of network formation to be triggered. Locations of the network formation events are assigned dependent on the spatial distribution of the soluble pool of filament precursors. Based on this modeling approach, simulation studies revealed that the architecture of keratin networks mostly depends on the balance between filament elongation and branching processes. The spatial distribution of network mesh size, which strongly influences the mechanical characteristics of filament networks, is modulated by lateral annealing processes. This mechanism which is a specific feature of intermediate filament networks appears to be a major and fast regulator of cell mechanics.

  2. Magnetic helicity and active filament configuration

    Science.gov (United States)

    Romano, P.; Zuccarello, F.; Poedts, S.; Soenen, A.; Zuccarello, F. P.

    2009-11-01

    Context: The role of magnetic helicity in active filament formation and destabilization is still under debate. Aims: Although active filaments usually show a sigmoid shape and a twisted configuration before and during their eruption, it is unclear which mechanism leads to these topologies. In order to provide an observational contribution to clarify these issues, we describe a filament evolution whose characteristics seem to be directly linked to the magnetic helicity transport in corona. Methods: We applied different methods to determine the helicity sign and the chirality of the filament magnetic field. We also computed the magnetic helicity transport rate at the filament footpoints. Results: All the observational signatures provided information on the positive helicity and sinistral chirality of the flux rope containing the filament material: its forward S shape, the orientation of its barbs, the bright and dark threads at 195 Å. Moreover, the magnetic helicity transport rate at the filament footpoints showed a clear accumulation of positive helicity. Conclusions: The study of this event showed a correspondence between several signatures of the sinistral chirality of the filament and several evidences of the positive magnetic helicity of the filament magnetic field. We also found that the magnetic helicity transported along the filament footpoints showed an increase just before the change of the filament shape observed in Hα images. We argued that the photospheric regions where the filament was rooted might be the preferential ways where the magnetic helicity was injected along the filament itself and where the conditions to trigger the eruption were yielded.

  3. Differential proteomics and physiology of Pseudomonas putida KT2440 under filament-inducing conditions

    Directory of Open Access Journals (Sweden)

    Crabbé Aurélie

    2012-11-01

    Full Text Available Abstract Background Pseudomonas putida exerts a filamentous phenotype in response to environmental stress conditions that are encountered during its natural life cycle. This study assessed whether P. putida filamentation could confer survival advantages. Filamentation of P. putida was induced through culturing at low shaking speed and was compared to culturing in high shaking speed conditions, after which whole proteomic analysis and stress exposure assays were performed. Results P. putida grown in filament-inducing conditions showed increased resistance to heat and saline stressors compared to non-filamented cultures. Proteomic analysis showed a significant metabolic change and a pronounced induction of the heat shock protein IbpA and recombinase RecA in filament-inducing conditions. Our data further indicated that the associated heat shock resistance, but not filamentation, was dependent of RecA. Conclusions This study provides insights into the altered metabolism of P. putida in filament-inducing conditions, and indicates that the formation of filaments could potentially be utilized by P. putida as a survival strategy in its hostile, recurrently changing habitat.

  4. Filament Winding. A Unified Approach

    NARCIS (Netherlands)

    Koussios, S.

    2004-01-01

    In this dissertation we have presented an overview and comprehensive treatment of several facets of the filament winding process. With the concepts of differential geometry and the theory of thin anisotropic shells of revolution, a parametric shape generator has been formulated for the design

  5. Towards filament free semiconductor lasers

    DEFF Research Database (Denmark)

    McInerney, John; O'Brien, Peter; Skovgaard, Peter M. W.

    2000-01-01

    We outline physical models and simulations for suppression of self-focusing and filamentation in large aperture semiconductor lasers. The principal technical objective is to generate multi-watt CW or quasi-CW outputs with nearly diffraction limited beams, suitable for long distance free space...

  6. Capillary thinning of polymeric filaments

    DEFF Research Database (Denmark)

    Kolte, Mette Irene; Szabo, Peter

    1999-01-01

    The capillary thinning of filaments of a Newtonian polybutene fluid and a viscoelastic polyisobutylene solution are analyzed experimentally and by means of numerical simulation. The experimental procedure is as follows. Initially, a liquid sample is placed between two cylindrical plates. Then...

  7. Capillary thinning of polymeric filaments

    DEFF Research Database (Denmark)

    Kolte, Mette Irene; Szabo, Peter; Hassager, Ole

    1998-01-01

    The capillary thinning of a polymeric filament is analysed experimentally as well as by means of numerical simulation. The experimental procedure is as follows. Initially a liquid sample is kept between two cylindrical plates. Then the bottom plate is lowered under gravity to yield a given strain...

  8. Various Barbs in Solar Filaments

    Science.gov (United States)

    Filippov, Boris

    2017-07-01

    Interest to lateral details of the solar filament shape named barbs, motivated by their relationship to filament chirality and helicity, showed their different orientation relative to the expected direction of the magnetic field. While the majority of barbs are stretched along the field, some barbs seem to be transversal to it and are referred to as anomalous barbs. We analyse the deformation of helical field lines by a small parasitic polarity using a simple flux rope model with a force-free field. A rather small and distant source of parasitic polarity stretches the bottom parts of the helical lines in its direction creating a lateral extension of dips below the flux-rope axis. They can be considered as normal barbs of the filament. A stronger and closer source of parasitic polarity makes the flux-rope field lines to be convex below its axis and creates narrow and deep dips near its position. As a result, the narrow structure, with thin threads across it, is formed whose axis is nearly perpendicular to the field. The structure resembles an anomalous barb. Hence, the presence of anomalous barbs does not contradict the flux-rope structure of a filament.

  9. Transient filament stretching rheometer II

    DEFF Research Database (Denmark)

    Kolte, Mette Irene; Rasmussen, Henrik K.; Hassager, Ole

    1997-01-01

    The Lagrangian sspecification is used to simulate the transient stretching filament rheometer. Simulations are performed for dilute PIB-solutions modeled as a four mode Oldroyd-B fluid and a semidilute PIB-solution modeled as a non-linear single integral equation. The simulations are compared...

  10. Towards filament free semiconductor lasers

    DEFF Research Database (Denmark)

    McInerney, John; O'Brien, Peter; Skovgaard, Peter M. W.

    2000-01-01

    We outline physical models and simulations for suppression of self-focusing and filamentation in large aperture semiconductor lasers. The principal technical objective is to generate multi-watt CW or quasi-CW outputs with nearly diffraction limited beams, suitable for long distance free space...... propagation structures in lasers and amplifiers which suppress lateral reflections....

  11. Positrusion Filament Recycling System, Phase I

    Data.gov (United States)

    National Aeronautics and Space Administration — TUI proposes a novel process to produce 3d printer feedstock filament out of scrap ABS on the ISS. Currently the plastic filament materials that most 3d printers use...

  12. Femtosecond Laser Filamentation for Atmospheric Sensing

    OpenAIRE

    Huai Liang Xu; See Leang Chin

    2010-01-01

    Powerful femtosecond laser pulses propagating in transparent materials result in the formation of self-guided structures called filaments. Such filamentation in air can be controlled to occur at a distance as far as a few kilometers, making it ideally suited for remote sensing of pollutants in the atmosphere. On the one hand, the high intensity inside the filaments can induce the fragmentation of all matters in the path of filaments, resulting in the emission of characteristic fluorescence sp...

  13. Filament Winding Of Carbon/Carbon Structures

    Science.gov (United States)

    Jacoy, Paul J.; Schmitigal, Wesley P.; Phillips, Wayne M.

    1991-01-01

    Improved method of winding carbon filaments for carbon/carbon composite structures less costly and labor-intensive, also produces more consistent results. Involves use of roller squeegee to ensure filaments continuously wet with resin during winding. Also involves control of spacing and resin contents of plies to obtain strong bonds between carbon filaments and carbon matrices. Lends itself to full automation and involves use of filaments and matrix-precursor resins in their simplest forms, thereby reducing costs.

  14. Galalpha1-->4Gal-glycans are expressed on myofibrillar associated proteins

    DEFF Research Database (Denmark)

    Kirkeby, S; Moe, D; Cläesson, M H

    1998-01-01

    NAcbeta- were used to detect terminal alpha-galactosylated glycoconjugates on muscle proteins. Electrotransfer of proteins, extracted from human masseter and biceps muscles, to nitrocellulose after polyacrylamide gel electrophoresis (PAGE) and incubation with anti-Pk (CD77) consistently showed two bands......-fixed human muscle displayed a CD77 reaction with highest accumulation of reaction product at the periphery of the fibers. This may be explained by the presence of Pk glycoconjugates on intermediate filaments in muscle fibers. In preparations of cat masseter muscle proteins the antibodies against P1Pk...... indicates that glycans carrying Galalpha1-4Galbeta1- epitopes are expressed on myofibrillar associated proteins....

  15. Solar Filaments as Tracers of Subsurface Processes

    Indian Academy of Sciences (India)

    tribpo

    Filaments are clouds of relatively cool and dense gas in the solar atmosphere. ... First-tier filaments may be related to a peculiar feature of the solar dynamo. .... Still, an appeal to subsurface processes should be resisted, but surface motion models have been able to reproduce neither the pattern of filament field orientations ...

  16. Striation and convection in penumbral filaments

    NARCIS (Netherlands)

    Spruit, H.C.; Scharmer, G.B.; Löfdahl, M.G.

    2010-01-01

    Observations with the 1-m Swedish Solar Telescope of the flows seen in penumbral filaments are presented. Time sequences of bright filaments show overturning motions strikingly similar to those seen along the walls of small isolated structures in the active regions. The filaments show outward

  17. Solar Filaments as Tracers of Subsurface Processes

    Indian Academy of Sciences (India)

    tribpo

    according to which, probably all the magnetic flux that emerges into the photosphere is twisted. Twisted flux forms sunspots, active regions (ARs) and filaments. The twist accumulates in filaments and coronal arcades. Eventually the accumulated, highly twisted fields become unstable and erupt. From a study of filament ...

  18. Effectors of Filamentous Plant Pathogens: Commonalities amid Diversity.

    Science.gov (United States)

    Franceschetti, Marina; Maqbool, Abbas; Jiménez-Dalmaroni, Maximiliano J; Pennington, Helen G; Kamoun, Sophien; Banfield, Mark J

    2017-06-01

    Fungi and oomycetes are filamentous microorganisms that include a diversity of highly developed pathogens of plants. These are sophisticated modulators of plant processes that secrete an arsenal of effector proteins to target multiple host cell compartments and enable parasitic infection. Genome sequencing revealed complex catalogues of effectors of filamentous pathogens, with some species harboring hundreds of effector genes. Although a large fraction of these effector genes encode secreted proteins with weak or no sequence similarity to known proteins, structural studies have revealed unexpected similarities amid the diversity. This article reviews progress in our understanding of effector structure and function in light of these new insights. We conclude that there is emerging evidence for multiple pathways of evolution of effectors of filamentous plant pathogens but that some families have probably expanded from a common ancestor by duplication and diversification. Conserved folds, such as the oomycete WY and the fungal MAX domains, are not predictive of the precise function of the effectors but serve as a chassis to support protein structural integrity while providing enough plasticity for the effectors to bind different host proteins and evolve unrelated activities inside host cells. Further effector evolution and diversification arise via short linear motifs, domain integration and duplications, and oligomerization. Copyright © 2017 American Society for Microbiology.

  19. Fine oral filaments in Paramecium: a biochemical and immunological analysis.

    Science.gov (United States)

    Clerot, J; Iftode, F; Budin, K; Jeanmaire-Wolf, R; Coffe, G; Fleury-Aubusson, A

    2001-01-01

    In Paramecium, several kinds of the oral networks of fine filaments are defined at the ultrastructural level. Using the sodium chloride-treated oral apparatus of Paramecium as an antigen to produce monoclonal antibodies, we have begun to identify the proteins constituting these networks. Immunoblotting showed that all positive antibodies were directed against three bands (70-, 75-and 83-kD), which corresponded to quantitatively minor components of the antigen; there was no antibody specific for the quantitatively major components (58- and 62-kD). Immunolocalization with four of these antibodies directed against one or several of these three bands showed that these proteins are components of the fine filaments supporting the oral area; a decoration of the basal bodies and the outer lattice was also observed on the cortex. Immunofluorescence on interphase cells suggested that the three proteins colocalized on the left side of the oral apparatus, whereas only the 70-kD band was detected on the right side. During division, the antigens of the antibodies were detected at different stages after oral basal body assembly. The antibodies cross-reacted with the tetrins, which are oral filament-forming proteins in Tetrahymena, demonstrating that tetrin-related proteins are quantitatively minor components of the oral and the somatic cytoskeleton of Paramecium.

  20. Elasticity of a Filament with Kinks

    Science.gov (United States)

    Razbin, Mohammadhosein

    2017-12-01

    Using the wormlike chain model, we analytically study the elasticity of a filament with kinks. We calculate the position probability density function and the force constant of a kinked filament with a general kink angle. Then, using the mathematical induction, we obtain the positional-orientational probability density function of a filament with regular kinks. For this filament, we compute the force constant in two different directions. In longitudinal direction of the filament, the force constant is proportional to the inverse of the number of the segments, i.e., 1 / m, while in transverse direction, it is proportional to 1/m^3.

  1. UNUSUAL FILAMENTS INSIDE THE UMBRA

    Energy Technology Data Exchange (ETDEWEB)

    Kleint, L. [High Altitude Observatory/NCAR, P.O. Box 3000, Boulder, CO 80307 (United States); Sainz Dalda, A., E-mail: kleintl@ucar.edu [Stanford-Lockheed Institute for Space Research, Stanford University, HEPL, 466 Via Ortega, Stanford, CA 94305 (United States)

    2013-06-10

    We analyze several unusual filamentary structures which appeared in the umbra of one of the sunspots in AR 11302. They do not resemble typical light bridges in morphology or in evolution. We analyze data from SDO/HMI to investigate their temporal evolution, Hinode/SP for photospheric inversions, IBIS for chromospheric imaging, and SDO/AIA for the overlying corona. Photospheric inversions reveal a horizontal, inverse Evershed flow along these structures, which we call umbral filaments. Chromospheric images show brightenings and energy dissipation, while coronal images indicate that bright coronal loops seem to end in these umbral filaments. These rapidly evolving features do not seem to be common, and are possibly related to the high flare-productivity of the active region. Their analysis could help to understand the complex evolution of active regions.

  2. The Apis mellifera Filamentous Virus Genome

    Directory of Open Access Journals (Sweden)

    Laurent Gauthier

    2015-07-01

    Full Text Available A complete reference genome of the Apis mellifera Filamentous virus (AmFV was determined using Illumina Hiseq sequencing. The AmFV genome is a double stranded DNA molecule of approximately 498,500 nucleotides with a GC content of 50.8%. It encompasses 247 non-overlapping open reading frames (ORFs, equally distributed on both strands, which cover 65% of the genome. While most of the ORFs lacked threshold sequence alignments to reference protein databases, twenty-eight were found to display significant homologies with proteins present in other large double stranded DNA viruses. Remarkably, 13 ORFs had strong similarity with typical baculovirus domains such as PIFs (per os infectivity factor genes: pif-1, pif-2, pif-3 and p74 and BRO (Baculovirus Repeated Open Reading Frame. The putative AmFV DNA polymerase is of type B, but is only distantly related to those of the baculoviruses. The ORFs encoding proteins involved in nucleotide metabolism had the highest percent identity to viral proteins in GenBank. Other notable features include the presence of several collagen-like, chitin-binding, kinesin and pacifastin domains. Due to the large size of the AmFV genome and the inconsistent affiliation with other large double stranded DNA virus families infecting invertebrates, AmFV may belong to a new virus family.

  3. Role of Tellurite Resistance Operon in Filamentous Growth of Yersinia pestis in Macrophages.

    Science.gov (United States)

    Ponnusamy, Duraisamy; Clinkenbeard, Kenneth D

    2015-01-01

    Yersinia pestis initiates infection by parasitism of host macrophages. In response to macrophage infections, intracellular Y. pestis can assume a filamentous cellular morphology which may mediate resistance to host cell innate immune responses. We previously observed the expression of Y. pestis tellurite resistance proteins TerD and TerE from the terZABCDE operon during macrophage infections. Others have observed a filamentous response associated with expression of tellurite resistance operon in Escherichia coli exposed to tellurite. Therefore, in this study we examine the potential role of Y. pestis tellurite resistance operon in filamentous cellular morphology during macrophage infections. In vitro treatment of Y. pestis culture with sodium tellurite (Na2TeO3) caused the bacterial cells to assume a filamentous phenotype similar to the filamentous phenotype observed during macrophage infections. A deletion mutant for genes terZAB abolished the filamentous morphologic response to tellurite exposure or intracellular parasitism, but without affecting tellurite resistance. However, a terZABCDE deletion mutant abolished both filamentous morphologic response and tellurite resistance. Complementation of the terZABCDE deletion mutant with terCDE, but not terZAB, partially restored tellurite resistance. When the terZABCDE deletion mutant was complemented with terZAB or terCDE, Y. pestis exhibited filamentous morphology during macrophage infections as well as while these complemented genes were being expressed under an in vitro condition. Further in E. coli, expression of Y. pestis terZAB, but not terCDE, conferred a filamentous phenotype. These findings support the role of Y. pestis terZAB mediation of the filamentous response phenotype; whereas, terCDE confers tellurite resistance. Although the beneficial role of filamentous morphological responses by Y. pestis during macrophage infections is yet to be fully defined, it may be a bacterial adaptive strategy to macrophage

  4. Lighting the universe with filaments.

    Science.gov (United States)

    Gao, Liang; Theuns, Tom

    2007-09-14

    The first stars in the universe form when chemically pristine gas heats as it falls into dark-matter potential wells, cools radiatively because of the formation of molecular hydrogen, and becomes self-gravitating. Using supercomputer simulations, we demonstrated that the stars' properties depend critically on the currently unknown nature of the dark matter. If the dark-matter particles have intrinsic velocities that wipe out small-scale structure, then the first stars form in filaments with lengths on the order of the free-streaming scale, which can be approximately 10(20) meters (approximately 3 kiloparsecs, corresponding to a baryonic mass of approximately 10(7) solar masses) for realistic "warm dark matter" candidates. Fragmentation of the filaments forms stars with a range of masses, which may explain the observed peculiar element abundance pattern of extremely metal-poor stars, whereas coalescence of fragments and stars during the filament's ultimate collapse may seed the supermassive black holes that lurk in the centers of most massive galaxies.

  5. Direct observation of current in type-I edge-localized-mode filaments on the ASDEX Upgrade tokamak.

    Science.gov (United States)

    Vianello, N; Naulin, V; Schrittwieser, R; Müller, H W; Zuin, M; Ionita, C; Rasmussen, J J; Mehlmann, F; Rohde, V; Cavazzana, R; Maraschek, M

    2011-03-25

    Magnetically confined plasmas in the high confinement regime are regularly subjected to relaxation oscillations, termed edge localized modes (ELMs), leading to large transport events. Present ELM theories rely on a combined effect of edge current and the edge pressure gradients which result in intermediate mode number (n≅10-15) structures (filaments) localized in the perpendicular plane and extended along the field lines. By detailed localized measurements of the magnetic field perturbation associated to type-I ELM filaments, it is shown that these filaments carry a substantial current.

  6. Characterization of a 65 kDa NIF in the nuclear matrix of the monocot Allium cepa that interacts with nuclear spectrin-like proteins.

    Science.gov (United States)

    Pérez-Munive, Clara; Blumenthal, Sonal S D; de la Espina, Susana Moreno Díaz

    2012-01-01

    Plant cells have a well organized nucleus and nuclear matrix, but lack orthologues of the main structural components of the metazoan nuclear matrix. Although data is limited, most plant nuclear structural proteins are coiled-coil proteins, such as the NIFs (nuclear intermediate filaments) in Pisum sativum that cross-react with anti-intermediate filament and anti-lamin antibodies, form filaments 6-12 nm in diameter in vitro, and may play the role of lamins. We have investigated the conservation and features of NIFs in a monocot species, Allium cepa, and compared them with onion lamin-like proteins. Polyclonal antisera against the pea 65 kDa NIF were used in 1D and 2D Western blots, ICM (imunofluorescence confocal microscopy) and IEM (immunoelectron microscopy). Their presence in the nuclear matrix was analysed by differential extraction of nuclei, and their association with structural spectrin-like proteins by co-immunoprecipitation and co-localization in ICM. NIF is a conserved structural component of the nucleus and its matrix in monocots with Mr and pI values similar to those of pea 65 kDa NIF, which localized to the nuclear envelope, perichromatin domains and foci, and to the nuclear matrix, interacting directly with structural nuclear spectrin-like proteins. Its similarities with some of the proteins described as onion lamin-like proteins suggest that they are highly related or perhaps the same proteins.

  7. [The structure of thick filaments on longitudinal sections of rabbit psoas muscle].

    Science.gov (United States)

    Podlubnaia, Z A; Latsabidze, I L; Lednev, V V

    1989-01-01

    By means of electron microscopy the longitudinal sections of chemically skinned fibres of rigorised rabbit psoas muscle have been examined at pH of rigorising solutions equal to 6, 7, 8 (I = 0.125) and ionic strengths equal to 0.04, 0.125, 0.34 (pH 7.0). It has been revealed that at pH 6.0 the bands of minor proteins localization in A-disks were seen very distinctly, while at pH 7.0 and I = 0.125 these bands can be revealed only by means of antibody labelling technique. At the ionic strength of 0.34 (pH 7.0) the periodicity of 14.3 nm in thick filaments was clearly observed, which was determined by packing of the myosin rods into the filament shaft and of the myosin heads (cross-bridges) on the filament surface. The number of cross-bridge rows in the filament equals 102. A new scheme of myosin cross-bridge distribution in thick filaments of rabbit psoas muscle has been suggested according to which two rows of cross-bridges at each end of a thick filament are absent. The filament length equals 1.64 +/- 0.01 micron. It has been shown that the length of thick filament as well as the structural organization of their end regions in rabbit psoas muscle and frog sartorius one are different.

  8. Cholera toxin can catalyze ADP-ribosylation of cytoskeletal proteins

    International Nuclear Information System (INIS)

    Kaslow, H.R.; Groppi, V.E.; Abood, M.E.; Bourne, H.R.

    1981-01-01

    Cholera toxin catalyzes transfer of radiolabel from [ 32 P]NAD + to several peptides in particulate preparations of human foreskin fibroblasts. Resolution of these peptides by two-dimensional gel electrophoresis allowed identification of two peptides of M/sub r/ = 42,000 and 52,000 as peptide subunits of a regulatory component of adenylate cyclase. The radiolabeling of another group of peptides (M/sub r/ = 50,000 to 65,000) suggested that cholera toxin could catalyze ADP-ribosylation of cytoskeletal proteins. This suggestion was confirmed by showing that incubation with cholera toxin and [ 32 P]NAD + caused radiolabeling of purified microtubule and intermediate filament proteins

  9. Differential assembly of alpha- and gamma-filagenins into thick filaments in Caenorhabditis elegans

    Science.gov (United States)

    Liu, F.; Ortiz, I.; Hutagalung, A.; Bauer, C. C.; Cook, R. G.; Epstein, H. F.

    2000-01-01

    Muscle thick filaments are highly organized supramolecular assemblies of myosin and associated proteins with lengths, diameters and flexural rigidities characteristic of their source. The cores of body wall muscle thick filaments of the nematode Caenorhabditis elegans are tubular structures of paramyosin sub-filaments coupled by filagenins and have been proposed to serve as templates for the assembly of native thick filaments. We have characterized alpha- and gamma-filagenins, two novel proteins of the cores with calculated molecular masses of 30,043 and 19,601 and isoelectric points of 10.52 and 11.49, respectively. Western blot and immunoelectron microscopy using affinity-purified antibodies confirmed that the two proteins are core components. Immunoelectron microscopy of the cores revealed that they assemble with different periodicities. Immunofluorescence microscopy showed that alpha-filagenin is localized in the medial regions of the A-bands of body wall muscle cells whereas gamma-filagenin is localized in the flanking regions, and that alpha-filagenin is expressed in 1.5-twofold embryos while gamma-filagenin becomes detectable only in late vermiform embryos. The expression of both proteins continues throughout later stages of development. C. elegans body wall muscle thick filaments of these developmental stages have distinct lengths. Our results suggest that the differential assembly of alpha- and gamma-filagenins into thick filaments of distinct lengths may be developmentally regulated.

  10. Decidable and undecidable arithmetic functions in actin filament networks

    Science.gov (United States)

    Schumann, Andrew

    2018-01-01

    The plasmodium of Physarum polycephalum is very sensitive to its environment, and reacts to stimuli with appropriate motions. Both the sensory and motor stages of these reactions are explained by hydrodynamic processes, based on fluid dynamics, with the participation of actin filament networks. This paper is devoted to actin filament networks as a computational medium. The point is that actin filaments, with contributions from many other proteins like myosin, are sensitive to extracellular stimuli (attractants as well as repellents), and appear and disappear at different places in the cell to change aspects of the cell structure—e.g. its shape. By assembling and disassembling actin filaments, some unicellular organisms, like Amoeba proteus, can move in response to various stimuli. As a result, these organisms can be considered a simple reversible logic gate—extracellular signals being its inputs and motions its outputs. In this way, we can implement various logic gates on amoeboid behaviours. These networks can embody arithmetic functions within p-adic valued logic. Furthermore, within these networks we can define the so-called diagonalization for deducing undecidable arithmetic functions.

  11. A rapid PCR-based approach for molecular identification of filamentous fungi.

    Science.gov (United States)

    Chen, Yuanyuan; Prior, Bernard A; Shi, Guiyang; Wang, Zhengxiang

    2011-08-01

    In this study, a novel rapid and efficient DNA extraction method based on alkaline lysis, which can deal with a large number of filamentous fungal isolates in the same batch, was established. The filamentous fungal genomic DNA required only 20 min to prepare and can be directly used as a template for PCR amplification. The amplified internal transcribed spacer regions were easy to identify by analysis. The extracted DNA also can be used to amplify other protein-coding genes for fungal identification. This method can be used for rapid systematic identification of filamentous fungal isolates.

  12. Nonlinear Force-free Field Extrapolation of a Coronal Magnetic Flux Rope Supporting a Large-scale Solar Filament from a Photospheric Vector Magnetogram

    Science.gov (United States)

    Jiang, Chaowei; Wu, S. T.; Feng, Xueshang; Hu, Qiang

    2014-05-01

    Solar filaments are commonly thought to be supported in magnetic dips, in particular, in those of magnetic flux ropes (FRs). In this Letter, based on the observed photospheric vector magnetogram, we implement a nonlinear force-free field (NLFFF) extrapolation of a coronal magnetic FR that supports a large-scale intermediate filament between an active region and a weak polarity region. This result is a first, in the sense that current NLFFF extrapolations including the presence of FRs are limited to relatively small-scale filaments that are close to sunspots and along main polarity inversion lines (PILs) with strong transverse field and magnetic shear, and the existence of an FR is usually predictable. In contrast, the present filament lies along the weak-field region (photospheric field strength barbs very well, which strongly supports the FR-dip model for filaments. The filament is stably sustained because the FR is weakly twisted and strongly confined by the overlying closed arcades.

  13. Part I---Evaluating Effects of Oligomer Formation on Cytochrome P450 2C9 Electron Transfer and Drug Metabolism, Part II---Utilizing Molecular Modeling Techniques to Study the Src-Interacting Proteins Actin Filament Associated Protein of 110 kDa (AFAP-110) and Cortactin

    Science.gov (United States)

    Jett, John Edward, Jr.

    The dissertation has been divided into two parts to accurately reflect the two distinct areas of interest pursued during my matriculation in the School of Pharmacy at West Virginia University. In Part I, I discuss research probing the nature of electron transfer in the Cytochrome P450 family of proteins, a group of proteins well-known for their role in drug metabolism. In Part II, I focus on in silico and in vitro work developed in concert to probe protein structure and protein-protein interactions involved in actin filament reorganization and cellular motility. Part I. Cytochrome P450s (P450s) are an important class of enzymes known to metabolize a variety of endogenous and xenobiotic compounds. P450s are most commonly found in liver and intestinal endothelial cells and are responsible for the metabolism of approximately 75% of pharmaceutical drugs on the market. CYP2C9---one of the six major P450 isoforms---is responsible for ˜20% of drug metabolism. Elucidation of the factors that affect in vitro drug metabolism is crucial to the accurate prediction of in vivo drug metabolism kinetics. Currently, the two major techniques for studying in vitro drug metabolism are solution-based. However, it is known that the results of solution-based studies can vary from in vivo drug metabolism. One reason suggested to account for this variation is the state of P450 oligomer formation in solution compared to the in vivo environment, where P450s are membrane-bound. To understand the details of how oligomer formation affects in vitro drug metabolism, it is imperative that techniques be developed which will allow for the unequivocal control of oligomer formation without altering other experimental parameters. Our long term goal of this research is to develop methods to more accurately predict in vivo drug metabolism from in vitro data. This section of the dissertation will discuss the development of a platform consisting of a doped silicon surface containing a large array of gold

  14. Femtosecond Laser Filamentation for Atmospheric Sensing

    Directory of Open Access Journals (Sweden)

    Huai Liang Xu

    2010-12-01

    Full Text Available Powerful femtosecond laser pulses propagating in transparent materials result in the formation of self-guided structures called filaments. Such filamentation in air can be controlled to occur at a distance as far as a few kilometers, making it ideally suited for remote sensing of pollutants in the atmosphere. On the one hand, the high intensity inside the filaments can induce the fragmentation of all matters in the path of filaments, resulting in the emission of characteristic fluorescence spectra (fingerprints from the excited fragments, which can be used for the identification of various substances including chemical and biological species. On the other hand, along with the femtosecond laser filamentation, white-light supercontinuum emission in the infrared to UV range is generated, which can be used as an ideal light source for absorption Lidar. In this paper, we present an overview of recent progress concerning remote sensing of the atmosphere using femtosecond laser filamentation.

  15. The Mysterious Case of the Missing Filaments

    Science.gov (United States)

    Alden, C. R.

    2016-12-01

    Coronal Mass Ejections, or CMEs, are large solar eruptions that can have major debilitating impacts on society. Typically, these eruptions have the three following key structures: the leading edge, the empty chamber known as the cavity, and the filament which often is the brightest part of the CME. When we can see all three structures clearly with a coronagraph, it is called a classic three-part CME, also referred to as a 'lightbulb' CME. According to current knowledge, when a CME erupts, a filament should also erupt or lift off the Sun in order to have the bright center within the CME. However, we do not always see a filament erupt at the surface, and yet we still get a 'filament' within the coronagraph CME. To better understand what might be occurring with these missing filaments, we looked at three-part CMEs using the SOHO LASCO CME Catalog and filaments from the SDO AIA Filament Catalog in order to create a list of 50 CMEs without a listed filament erupting at the surface. For those CMEs without filaments in the list we closely inspected the AIA images for evidence of filament eruption. To ensure that there were no filaments past the limb of the Sun, we used data from the STEREO-A and STEREO-B spacecraft's to look at the Sun from other angles. We have found numerous events where no filament erupts from the surface, but we still see the classic three-part CME. We believe this may be due to an optical illusion occurring from the twisting of the flux rope.

  16. Chaperonin filaments: their formation and an evaluation of methods for studying them.

    Science.gov (United States)

    Yaoi, T; Kagawa, H K; Trent, J D

    1998-08-01

    Chaperonins are multisubunit protein complexes that can be isolated from cells as high-molecular-weight structures that appear as double rings in the electron microscope. We recently discovered that chaperonin double rings isolated from the hyperthermophilic archaeon Sulfolobus shibatae, when incubated at physiological temperatures in the presence of ATP and Mg2+, stacked into filaments; we hypothesized that these filaments are related to filaments seen inside S. shibatae cells and that chaperonins exist as filaments in vivo (J. D. Trent et al., 1997, Proc. Natl. Acad. Sci. USA 94, 5383-5388). This paper elucidates the conditions under which we have observed S. shibatae chaperonins to form filaments and evaluates native polyacrylamide gel electrophoresis (PAGE), TEM, spectrophotometry, and centrifugation as methods for studying these filaments. We observed that in the presence of Mg2+ combined with ATP, ADP, ATPgammaS, or GTP, native PAGE indicated that chaperonin subunits assembled into double rings and that the conformation of these double rings was effected by nucleotide binding, but we saw no indication of chaperonin filament formation. Under these same conditions, however, TEM, spectroscopy, and centrifugation methods indicated that chaperonin subunits and double rings had assembled into filaments. We determined that this discrepancy in the representation of the chaperonin structure was due to the native PAGE method itself. When we exposed chaperonin filaments to the electrophoretic field used in native PAGE, the filaments dissociated into double rings. This suggests that TEM, spectrophotometry, and centrifugation are the preferred methods for studying the higher-order structures of chaperonins, which are likely to be of biological significance. Copyright 1998 Academic Press.

  17. Assembly of Superparamagnetic Filaments in External Field.

    Science.gov (United States)

    Wei, Jiachen; Song, Fan; Dobnikar, Jure

    2016-09-13

    We present a theoretical and simulation study of anchored magneto-elastic filaments in external magnetic field. The filaments are composed of a mixture of superparamagnetic and nonmagnetic colloidal beads interlinked with elastic springs. We explore the steady-state structures of filaments with various composition and bending rigidity subject to external magnetic field parallel to the surface. The interplay of elastic and induced magnetic interactions results in a rich phase behavior with morphologies reminiscent of macromolecular folding: bent filaments, loops, sheets, helicoids, and other collapsed structures. Our results provide new insights into the design of hierarchically assembled supramolecular structures with controlled response to external stimuli.

  18. Stable Translocation Intermediates Jam Global Protein Export in Plasmodium falciparum Parasites and Link the PTEX Component EXP2 with Translocation Activity.

    Science.gov (United States)

    Mesén-Ramírez, Paolo; Reinsch, Ferdinand; Blancke Soares, Alexandra; Bergmann, Bärbel; Ullrich, Ann-Katrin; Tenzer, Stefan; Spielmann, Tobias

    2016-05-01

    Protein export is central for the survival and virulence of intracellular P. falciparum blood stage parasites. To reach the host cell, exported proteins cross the parasite plasma membrane (PPM) and the parasite-enclosing parasitophorous vacuole membrane (PVM), a process that requires unfolding, suggestive of protein translocation. Components of a proposed translocon at the PVM termed PTEX are essential in this phase of export but translocation activity has not been shown for the complex and questions have been raised about its proposed membrane pore component EXP2 for which no functional data is available in P. falciparum. It is also unclear how PTEX mediates trafficking of both, soluble as well as transmembrane proteins. Taking advantage of conditionally foldable domains, we here dissected the translocation events in the parasite periphery, showing that two successive translocation steps are needed for the export of transmembrane proteins, one at the PPM and one at the PVM. Our data provide evidence that, depending on the length of the C-terminus of the exported substrate, these steps occur by transient interaction of the PPM and PVM translocon, similar to the situation for protein transport across the mitochondrial membranes. Remarkably, we obtained constructs of exported proteins that remained arrested in the process of being translocated across the PVM. This clogged the translocation pore, prevented the export of all types of exported proteins and, as a result, inhibited parasite growth. The substrates stuck in translocation were found in a complex with the proposed PTEX membrane pore component EXP2, suggesting a role of this protein in translocation. These data for the first time provide evidence for EXP2 to be part of a translocating entity, suggesting that PTEX has translocation activity and provide a mechanistic framework for the transport of soluble as well as transmembrane proteins from the parasite boundary into the host cell.

  19. Exploring the Secretomes of Microbes and Microbial Communities Using Filamentous Phage Display

    Directory of Open Access Journals (Sweden)

    Dragana eGagic

    2016-04-01

    Full Text Available Microbial surface and secreted proteins (the secretome contain a large number of proteins that interact with other microbes, host and/or environment. These proteins are exported by the coordinated activities of the protein secretion machinery present in the cell. A group of phage, called filamentous phage, have the ability to hijack the cellular protein secretion machinery in order to amplify and assemble via a secretion-like process. This ability has been harnessed in the use of filamentous phage of Escherichia coli in biotechnology applications, including screening large libraries of variants for binding to bait of interest, from tissues in vivo to pure proteins or even inorganic substrates. In this review we discuss the roles of secretome proteins in pathogenic and non-pathogenic bacteria and corresponding secretion pathways. We describe the basics of phage display technology and its variants applied to discovery of bacterial proteins that have functions of interest for bacterial colonization and pathogenesis, through filamentous phage display library screening. Published literature also shows that phage display is suitable for secretome protein display as a tool for identification immunogenic peptides and can be used for discovery of vaccine candidates. Secretome selection aided by next-generation sequence analysis can also be used for selective display of the secretome at a microbial community scale, the latter revealing the richness of secretome functions of interest and surprising versatility in filamentous phage display of secretome proteins from large number of Gram-negative as well as Gram-positive bacteria and archaea.

  20. Differential scanning calorimetry study of glycerinated rabbit psoas muscle fibres in intermediate state of ATP hydrolysis

    Directory of Open Access Journals (Sweden)

    Farkas Nelli

    2007-06-01

    Full Text Available Abstract Background Thermal denaturation experiments were extended to study the thermal behaviour of the main motor proteins (actin and myosin in their native environment in striated muscle fibres. The interaction of actin with myosin in the highly organized muscle structure is affected by internal forces; therefore their altered conformation and interaction may differ from those obtained in solution. The energetics of long functioning intermediate states of ATP hydrolysis cycle was studied in muscle fibres by differential scanning calorimetry (DSC. Results SETARAM Micro DSC-II was used to monitor the thermal denaturation of the fibre system in rigor and in the presence of nucleotide and nucleotide analogues. The AM.ADP.Pi state of the ATP hydrolysis cycle has a very short lifetime therefore, we mimicked the different intermediate states with AMP.PNP and/or inorganic phosphate analogues Vi and AlF4 or BeFx. Studying glycerol-extracted muscle fibres from the rabbit psoas muscle by DSC, three characteristic thermal transitions were detected in rigor. The thermal transitions can be assigned to myosin heads, myosin rods and actin with transition temperatures (Tm of 52.9 ± 0.7°C, 57.9 ± 0.7°C, 63.7 ± 1.0°C. In different intermediate states of the ATP hydrolysis mimicked by nucleotide analogues a fourth thermal transition was also detected which is very likely connected with nucleotide binding domain of myosin and/or actin filaments. This transition temperature Tm4 depended on the mimicked intermediate states, and varied in the range of 66°C – 77°C. Conclusion According to DSC measurements, strongly and weakly binding states of myosin to actin were significantly different. In the presence of ADP only a moderate change of the DSC pattern was detected in comparison with rigor, whereas in ADP.Pi state trapped by Vi, AlF4 or BeFx a remarkable stabilization was detected on the myosin head and actin filament which is reflected in a 3.0 – 10.0

  1. Subcutaneous implants of polypropylene filaments.

    Science.gov (United States)

    Liebert, T C; Chartoff, R P; Cosgrove, S L; McCuskey, R S

    1976-11-01

    Extruded filaments of unmodified polypropylene (PP) with and without antioxidant were implanted subcutaneously in hamsters in order to determine their rate of degradation. Specimens were removed periodically during a 5 month test period and analyzed by infrared spectroscopy and dynamic mechanical testing. The analyses show that degradation beigns to occur after only a few days. Although the reaction sequence is not known, several factors suggest that the in vivo degradation process is similar to autoxidation which occurs in air or oxygen. The infrared data indicate that the hydroxyl content of the implants increases at a rate of 0.061 mg/g polypropylene per day during the initiation phase of the reaction. An induction time of 108 days was extablished. Carbonyl bonds appear after an implantation time of 50--90 days and increase therafter. Mechanical tests indicate a decrease in the dynamic loss tangent, tan delta, during the first month of implantation for unmodified polypropylene. No change in the infrared spectra or tan delta was observed, however, for implants containing an antioxidant. Thus, it is apparent that polypropylene filaments implanted subcutaneously in hamsters degrade by an oxidation process which is retarded effectively by using an antioxidant. While the findings reported are specific to subcutaneous polypropylene implants, they suggest that degradation of other systems may involve similar processes. This notion suggests directions for further research on increasing the in vivo stability of synthetic polymers. Long-term effects of polymer implantation upon tissue were not studied in this work.

  2. Structure determination of helical filaments by solid-state NMR spectroscopy

    Science.gov (United States)

    Ahmed, Mumdooh; Spehr, Johannes; König, Renate; Lünsdorf, Heinrich; Rand, Ulfert; Lührs, Thorsten; Ritter, Christiane

    2016-01-01

    The controlled formation of filamentous protein complexes plays a crucial role in many biological systems and represents an emerging paradigm in signal transduction. The mitochondrial antiviral signaling protein (MAVS) is a central signal transduction hub in innate immunity that is activated by a receptor-induced conversion into helical superstructures (filaments) assembled from its globular caspase activation and recruitment domain. Solid-state NMR (ssNMR) spectroscopy has become one of the most powerful techniques for atomic resolution structures of protein fibrils. However, for helical filaments, the determination of the correct symmetry parameters has remained a significant hurdle for any structural technique and could thus far not be precisely derived from ssNMR data. Here, we solved the atomic resolution structure of helical MAVSCARD filaments exclusively from ssNMR data. We present a generally applicable approach that systematically explores the helical symmetry space by efficient modeling of the helical structure restrained by interprotomer ssNMR distance restraints. Together with classical automated NMR structure calculation, this allowed us to faithfully determine the symmetry that defines the entire assembly. To validate our structure, we probed the protomer arrangement by solvent paramagnetic resonance enhancement, analysis of chemical shift differences relative to the solution NMR structure of the monomer, and mutagenesis. We provide detailed information on the atomic contacts that determine filament stability and describe mechanistic details on the formation of signaling-competent MAVS filaments from inactive monomers. PMID:26733681

  3. Filament poisoning at typical carbon nanotube deposition conditions by hot-filament CVD

    CSIR Research Space (South Africa)

    Oliphant, CJ

    2009-05-01

    Full Text Available This paper reports on the poisoning of tungsten filaments during the hot-filament chemical vapour deposition process at typical carbon nanotube (CNT) deposition conditions and filament temperatures ranging from 1400 to 2000 °C. The morphological...

  4. Identification of proteins from 4200-year-old skin and muscle tissue biopsies from ancient Egyptian mummies of the first intermediate period shows evidence of acute inflammation and severe immune response.

    Science.gov (United States)

    Jones, Jana; Mirzaei, Mehdi; Ravishankar, Prathiba; Xavier, Dylan; Lim, Do Seon; Shin, Dong Hoon; Bianucci, Raffaella; Haynes, Paul A

    2016-10-28

    We performed proteomics analysis on four skin and one muscle tissue samples taken from three ancient Egyptian mummies of the first intermediate period, approximately 4200 years old. The mummies were first dated by radiocarbon dating of the accompany-\\break ing textiles, and morphologically examined by scanning electron microscopy of additional skin samples. Proteins were extracted, separated on SDS-PAGE (sodium dodecyl sulfate polyacrylamide gel electrophoresis) gels, and in-gel digested with trypsin. The resulting peptides were analysed using nanoflow high-performance liquid chromatography-mass spectrometry. We identified a total of 230 unique proteins from the five samples, which consisted of 132 unique protein identifications. We found a large number of collagens, which was confirmed by our microscopy data, and is in agreement with previous studies showing that collagens are very long-lived. As expected, we also found a large number of keratins. We identified numerous proteins that provide evidence of activation of the innate immunity system in two of the mummies, one of which also contained proteins indicating severe tissue inflammation, possibly indicative of an infection that we can speculate may have been related to the cause of death.This article is part of the themed issue 'Quantitative mass spectrometry'. © 2016 The Author(s).

  5. A Statistical Study of Solar Filament Eruptions

    Science.gov (United States)

    Schanche, Nicole; Aggarwal, Ashna; Reeves, Kathy; Kempton, Dustin James; Angryk, Rafal

    2016-05-01

    Solar filaments are cool, dark channels of partially-ionized plasma that lie above the chromosphere. Their structure follows the neutral line between local regions of opposite magnetic polarity. Previous research (e.g. Schmieder et al. 2013, McCauley et al. 2015) has shown a positive correlation (70-80%) between the occurrence of filament eruptions and coronal mass ejections (CME’s). In this study, we attempt to use properties of the filament in order to predict whether or not a given filament will erupt. This prediction would help to better predict the occurrence of an oncoming CME. To track the evolution of a filament over time, a spatio-temporal algorithm that groups separate filament instances from the Heliophysics Event Knowledgebase (HEK) into filament tracks was developed. Filament features from the HEK metadata, such as length, chirality, and tilt are then combined with other physical features, such as the overlying decay index for two sets of filaments tracks - those that erupt and those that remain bound. Using statistical methods such as the Kolmogrov-Smirnov test and a Random Forest Classifier, we determine the effectiveness of the combined features in prediction. We conclude that there is significant overlap between the properties of filaments that erupt and those that do not, leading to predictions only ~5-10% above chance. However, the changes in features, such as a change in the filament's length over time, were determined to have the highest predictive power. We discuss the possible physical connections with the change in these features."This project has been supported by funding from the Division of Advanced Cyberinfrastructure within the Directorate for Computer and Information Science and Engineering, the Division of Astronomical Sciences within the Directorate for Mathematical and Physical Sciences, and the Division of Atmospheric and Geospace Sciences within the Directorate for Geosciences, under NSF award #1443061.”

  6. Actin Cytoskeleton and Golgi Involvement in Barley stripe mosaic virus Movement and Cell Wall Localization of Triple Gene Block Proteins

    Directory of Open Access Journals (Sweden)

    Hyoun-Sub Lim

    2013-03-01

    Full Text Available Barley stripe mosaic virus (BSMV induces massive actin filament thickening at the infection front of infected Nicotiana benthamiana leaves. To determine the mechanisms leading to actin remodeling, fluorescent protein fusions of the BSMV triple gene block (TGB proteins were coexpressed in cells with the actin marker DsRed: Talin. TGB ectopic expression experiments revealed that TGB3 is a major elicitor of filament thickening, that TGB2 resulted in formation of intermediate DsRed:Talin filaments, and that TGB1 alone had no obvious effects on actin filament structure. Latrunculin B (LatB treatments retarded BSMV cell-to-cell movement, disrupted actin filament organization, and dramatically decreased the proportion of paired TGB3 foci appearing at the cell wall (CW. BSMV infection of transgenic plants tagged with GFP-KDEL exhibited membrane proliferation and vesicle formation that were especially evident around the nucleus. Similar membrane proliferation occurred in plants expressing TGB2 and/or TGB3, and DsRed: Talin fluorescence in these plants colocalized with the ER vesicles. TGB3 also associated with the Golgi apparatus and overlapped with cortical vesicles appearing at the cell periphery. Brefeldin A treatments disrupted Golgi and also altered vesicles at the CW, but failed to interfere with TGB CW localization. Our results indicate that actin cytoskeleton interactions are important in BSMV cell-to-cell movement and for CW localization of TGB3.

  7. Characterization of actin filament severing by actophorin from Acanthamoeba castellanii

    Science.gov (United States)

    1991-01-01

    Actophorin is an abundant 15-kD actinbinding protein from Acanthamoeba that is thought to form a nonpolymerizable complex with actin monomers and also to reduce the viscosity of polymerized actin by severing filaments (Cooper et al., 1986. J. Biol. Chem. 261:477-485). Homologous proteins have been identified in sea urchin, chicken, and mammalian tissues. Chemical crosslinking produces a 1:1 covalent complex of actin and actophorin. Actophorin and profilin compete for crosslinking to actin monomers. The influence of actophorin on the steady-state actin polymer concentration gave a Kd of 0.2 microM for the complex of actophorin with actin monomers. Several new lines of evidence, including assays for actin filament ends by elongation rate and depolymerization rate, show that actophorin severs actin filaments both at steady state and during spontaneous polymerization. This is confirmed by direct observation in the light microscope and by showing that the effects of actophorin on the low shear viscosity of polymerized actin cannot be explained by monomer sequestration. The severing activity of actophorin is strongly inhibited by stoichiometric concentrations of phalloidin or millimolar concentrations of inorganic phosphate. PMID:1757465

  8. Vimentin filament organization and stress sensing depend on its single cysteine residue and zinc binding

    Science.gov (United States)

    Pérez-Sala, Dolores; Oeste, Clara L.; Martínez, Alma E.; Carrasco, M. Jesús; Garzón, Beatriz; Cañada, F. Javier

    2015-01-01

    The vimentin filament network plays a key role in cell architecture and signalling, as well as in epithelial–mesenchymal transition. Vimentin C328 is targeted by various oxidative modifications, but its role in vimentin organization is not known. Here we show that C328 is essential for vimentin network reorganization in response to oxidants and electrophiles, and is required for optimal vimentin performance in network expansion, lysosomal distribution and aggresome formation. C328 may fulfil these roles through interaction with zinc. In vitro, micromolar zinc protects vimentin from iodoacetamide modification and elicits vimentin polymerization into optically detectable structures; in cells, zinc closely associates with vimentin and its depletion causes reversible filament disassembly. Finally, zinc transport-deficient human fibroblasts show increased vimentin solubility and susceptibility to disruption, which are restored by zinc supplementation. These results unveil a critical role of C328 in vimentin organization and open new perspectives for the regulation of intermediate filaments by zinc. PMID:26031447

  9. A First Approach to Filament Dynamics

    Science.gov (United States)

    Silva, P. E. S.; de Abreu, F. Vistulo; Simoes, R.; Dias, R. G.

    2010-01-01

    Modelling elastic filament dynamics is a topic of high interest due to the wide range of applications. However, it has reached a high level of complexity in the literature, making it unaccessible to a beginner. In this paper we explain the main steps involved in the computational modelling of the dynamics of an elastic filament. We first derive…

  10. Filamentous Growth in Eremothecium Fungi

    DEFF Research Database (Denmark)

    Oskarsson, Therese

    The fungal kingdom encompasses a diverse group of organisms some of which have a great impact on human lives, either as domesticated benefactors or as human and crop pathogens. Using the filamentous fungus Ashbya gossypii and its close relative Eremothecium cymbalariae as model organisms, this th...... of molecular tools for E. cymbalariae to enable a faster and more efficient approach for genetic comparisons between Eremothecium genus fungi.......-regulatory activity of AgGts1, the protein could have additional actin organizing properties. In the second and third part, this thesis addresses the use of A. gossypii and its relative E. cymbalariae as model organisms for filamentous growth. A series of assays analyzed the capability of Eremothecium genus fungi...

  11. Theory of Semiflexible Filaments and Networks

    Directory of Open Access Journals (Sweden)

    Fanlong Meng

    2017-02-01

    Full Text Available We briefly review the recent developments in the theory of individual semiflexible filaments, and of a crosslinked network of such filaments, both permanent and transient. Starting from the free energy of an individual semiflexible chain, models on its force-extension relation and other mechanical properties such as Euler buckling are discussed. For a permanently crosslinked network of filaments, theories on how the network responds to deformation are provided, with a focus on continuum approaches. Characteristic features of filament networks, such as nonlinear stress-strain relation, negative normal stress, tensegrity, and marginal stability are discussed. In the new area of transient filament network, where the crosslinks can be dynamically broken and re-formed, we show some recent attempts for understanding the dynamics of the crosslinks, and the related rheological properties, such as stress relaxation, yield stress and plasticity.

  12. Particles trajectories in magnetic filaments

    Energy Technology Data Exchange (ETDEWEB)

    Bret, A. [ETSI Industriales, Universidad de Castilla-La Mancha, 13071 Ciudad Real (Spain); Instituto de Investigaciones Energéticas y Aplicaciones Industriales, Campus Universitario de Ciudad Real, 13071 Ciudad Real (Spain)

    2015-07-15

    The motion of a particle in a spatially harmonic magnetic field is a basic problem involved, for example, in the mechanism of formation of a collisionless shock. In such settings, it is generally reasoned that particles entering a Weibel generated turbulence are trapped inside it, provided their Larmor radius in the peak field is smaller than the field coherence length. The goal of this work is to put this heuristic conclusion on firm ground by studying, both analytically and numerically, such motion. A toy model is analyzed, consisting of a relativistic particle entering a region of space occupied by a spatially harmonic field. The particle penetrates the magnetic structure in a direction aligned with the magnetic filaments. Although the conclusions are not trivial, the main result is confirmed.

  13. Polar patterns of driven filaments.

    Science.gov (United States)

    Schaller, Volker; Weber, Christoph; Semmrich, Christine; Frey, Erwin; Bausch, Andreas R

    2010-09-02

    The emergence of collective motion exhibited by systems ranging from flocks of animals to self-propelled microorganisms to the cytoskeleton is a ubiquitous and fascinating self-organization phenomenon. Similarities between these systems, such as the inherent polarity of the constituents, a density-dependent transition to ordered phases or the existence of very large density fluctuations, suggest universal principles underlying pattern formation. This idea is followed by theoretical models at all levels of description: micro- or mesoscopic models directly map local forces and interactions using only a few, preferably simple, interaction rules, and more macroscopic approaches in the hydrodynamic limit rely on the systems' generic symmetries. All these models characteristically have a broad parameter space with a manifold of possible patterns, most of which have not yet been experimentally verified. The complexity of interactions and the limited parameter control of existing experimental systems are major obstacles to our understanding of the underlying ordering principles. Here we demonstrate the emergence of collective motion in a high-density motility assay that consists of highly concentrated actin filaments propelled by immobilized molecular motors in a planar geometry. Above a critical density, the filaments self-organize to form coherently moving structures with persistent density modulations, such as clusters, swirls and interconnected bands. These polar nematic structures are long lived and can span length scales orders of magnitudes larger than their constituents. Our experimental approach, which offers control of all relevant system parameters, complemented by agent-based simulations, allows backtracking of the assembly and disassembly pathways to the underlying local interactions. We identify weak and local alignment interactions to be essential for the observed formation of patterns and their dynamics. The presented minimal polar-pattern-forming system

  14. Regulatory roles of microtubule-associated proteins in neuronal morphogenesis. Involvement of the extracellular matrix

    Directory of Open Access Journals (Sweden)

    Ramírez G.

    1999-01-01

    Full Text Available As a result of recent investigations, the cytoskeleton can be viewed as a cytoplasmic system of interconnected filaments with three major integrative levels: self-assembling macromolecules, filamentous polymers, e.g., microtubules, intermediate filaments and actin filaments, and supramolecular structures formed by bundles of these filaments or networks resulting from cross-bridges between these major cytoskeletal polymers. The organization of this biological structure appears to be sensitive to fine spatially and temporally dependent regulatory signals. In differentiating neurons, regulation of cytoskeleton organization is particularly relevant, and the microtubule-associated protein (MAP tau appears to play roles in the extension of large neuritic processes and axons as well as in the stabilization of microtubular polymers along these processes. Within this context, tau is directly involved in defining neuronal polarity as well as in the generation of neuronal growth cones. There is increasing evidence that elements of the extracellular matrix contribute to the control of cytoskeleton organization in differentiating neurons, and that these regulations could be mediated by changes in MAP activity. In this brief review, we discuss the possible roles of tau in mediating the effects of extracellular matrix components on the internal cytoskeletal arrays and its organization in growing neurons.

  15. Human Rad51 filaments on double- and single-stranded DNA : Correlating regular and irregular forms with recombination function

    NARCIS (Netherlands)

    Ristic, D.; Modesti, M.; Van der Heijden, T.; Van Noort, J.; Dekker, C.; Kanaar, R.; Wyman, C.

    Recombinase proteins assembled into helical filaments on DNA are believed to be the catalytic core of homologous recombination. The assembly, disassembly and dynamic rearrangements of this structure must drive the DNA strand exchange reactions of homologous recombination. The sensitivity of

  16. A Bipolar Spindle of Antiparallel ParM Filaments Drives Bacterial Plasmid Segregation

    DEFF Research Database (Denmark)

    Gayathri, P; Fujii, T; Møller-Jensen, Jakob

    2012-01-01

    To ensure their stable inheritance by daughter cells during cell division, bacterial low copy-number plasmids make simple DNA segregating machines that use an elongating protein filament between sister plasmids. In the ParMRC system of Escherichia coli R1 plasmid, ParM, an actin-like protein, forms...

  17. Filament Activation in Response to Magnetic Flux Emergence and Cancellation in Filament Channels

    Science.gov (United States)

    Li, Ting; Zhang, Jun; Ji, Haisheng

    2015-06-01

    We conducted a comparative analysis of two filaments that showed a quite different activation in response to the flux emergence within the filament channels. The observations from the Solar Dynamics Observatory (SDO) and Global Oscillation Network Group (GONG) were made to analyze the two filaments on 2013 August 17 - 20 (SOL2013-08-17) and September 29 (SOL2013-09-29). The first event showed that the main body of the filament was separated into two parts when an active region (AR) emerged with a maximum magnetic flux of about 6.4×1021 Mx underlying the filament. The close neighborhood and common direction of the bright threads in the filament and the open AR fan loops suggest a similar magnetic connectivity of these two flux systems. The equilibrium of the filament was not destroyed three days after the start of the emergence of the AR. To our knowledge, similar observations have never been reported before. In the second event, the emerging flux occurred nearby a barb of the filament with a maximum magnetic flux of 4.2×1020 Mx, about one order of magnitude lower than that of the first event. Two patches of parasitic polarity in the vicinity of the barb merged, then cancelled with nearby network fields. About 20 hours after the onset of the emergence, the filament erupted. Our findings imply that the location of emerging flux within the filament channel is probably crucial to filament evolution. If the flux emergence appears nearby the barbs, it is highly likely that the emerging flux and the filament magnetic fields will cancel, which may lead to the eruption of the filament. The comparison of the two events shows that the emergence of a small AR may still not be enough to disrupt the stability of a filament system, and the actual eruption only occurs after the flux cancellation sets in.

  18. Automatic Detect and Trace of Solar Filaments

    Science.gov (United States)

    Fang, Cheng; Chen, P. F.; Tang, Yu-hua; Hao, Qi; Guo, Yang

    We developed a series of methods to automatically detect and trace solar filaments in solar Hα images. The programs are able to not only recognize filaments and determine their properties, such as the position, the area and other relevant parameters, but also to trace the daily evolution of the filaments. For solar full disk Hα images, the method consists of three parts: first, preprocessing is applied to correct the original images; second, the Canny edge-detection method is used to detect the filaments; third, filament properties are recognized through the morphological operators. For each Hα filament and its barb features, we introduced the unweighted undirected graph concept and adopted Dijkstra shortest-path algorithm to recognize the filament spine; then, using polarity inversion line shift method for measuring the polarities in both sides of the filament to determine the filament axis chirality; finally, employing connected components labeling method to identify the barbs and calculating the angle between each barb and spine to indicate the barb chirality. Our algorithms are applied to the observations from varied observatories, including the Optical & Near Infrared Solar Eruption Tracer (ONSET) in Nanjing University, Mauna Loa Solar Observatory (MLSO) and Big Bear Solar Observatory (BBSO). The programs are demonstrated to be effective and efficient. We used our method to automatically process and analyze 3470 images obtained by MLSO from January 1998 to December 2009, and a butterfly diagram of filaments is obtained. It shows that the latitudinal migration of solar filaments has three trends in the Solar Cycle 23: The drift velocity was fast from 1998 to the solar maximum; after the solar maximum, it became relatively slow and after 2006, the migration became divergent, signifying the solar minimum. About 60% filaments with the latitudes larger than 50 degree migrate towards the Polar Regions with relatively high velocities, and the latitudinal migrating

  19. Lifetime of titanium filament at constant current

    International Nuclear Information System (INIS)

    Chou, T.S.; Lanni, C.

    1981-01-01

    Titanium Sublimation Pump (TSP) represents the most efficient and the least expensive method to produce Ultra High Vacuum (UHV) in storage rings. In ISABELLE, a proton storage accelerator under construction at Brookhaven National Laboratory, for example, TSP provides a pumping speed for hydrogen of > 2 x 10 6 l/s. Due to the finite life of titanium filaments, new filaments have to be switched in before the end of filament burn out, to ensure smooth operation of the accelerator. Therefore, several operational modes that can be used to activate the TSP were studied. The constant current mode is a convenient way of maintaining constant evaporating rate by increasing the power input while the filament diameter decreases as titanium evaporates. The filaments used in this experiment were standard Varian 916-0024 filaments made of Ti 85%, Mo 15% alloy. During their lifetime at a constant current of 48 amperes, the evaporation rate rose to a maximum at about 10% of their life and then flattened out to a constant value, 0.25 g/hr. The maximum evaporation rate occurs coincidently with the recrystallization of 74% Ti 26% Mo 2 from microstructure crystalline at higher titanium concentration to macrostructure crystalline at lower titanium concentration. As the macrocrystal grows, the slip plane develops at the grain boundary resulting in high resistance at the slip plane which will eventually cause the filament burn out due to local heating

  20. Filaments in simulations of molecular cloud formation

    Energy Technology Data Exchange (ETDEWEB)

    Gómez, Gilberto C.; Vázquez-Semadeni, Enrique [Centro de Radioastronomía y Astrofísica, Universidad Nacional Autónoma de México, Campus Morelia Apartado Postal 3-72, 58090 Morelia, Michoacán (Mexico)

    2014-08-20

    We report on the filaments that develop self-consistently in a new numerical simulation of cloud formation by colliding flows. As in previous studies, the forming cloud begins to undergo gravitational collapse because it rapidly acquires a mass much larger than the average Jeans mass. Thus, the collapse soon becomes nearly pressureless, proceeding along its shortest dimension first. This naturally produces filaments in the cloud and clumps within the filaments. The filaments are not in equilibrium at any time, but instead are long-lived flow features through which the gas flows from the cloud to the clumps. The filaments are long-lived because they accrete from their environment while simultaneously accreting onto the clumps within them; they are essentially the locus where the flow changes from accreting in two dimensions to accreting in one dimension. Moreover, the clumps also exhibit a hierarchical nature: the gas in a filament flows onto a main, central clump but other, smaller-scale clumps form along the infalling gas. Correspondingly, the velocity along the filament exhibits a hierarchy of jumps at the locations of the clumps. Two prominent filaments in the simulation have lengths ∼15 pc and masses ∼600 M {sub ☉} above density n ∼ 10{sup 3} cm{sup –3} (∼2 × 10{sup 3} M {sub ☉} at n > 50 cm{sup –3}). The density profile exhibits a central flattened core of size ∼0.3 pc and an envelope that decays as r {sup –2.5} in reasonable agreement with observations. Accretion onto the filament reaches a maximum linear density rate of ∼30 M {sub ☉} Myr{sup –1} pc{sup –1}.

  1. Thermal and Chemical Evolution of Collapsing Filaments

    Energy Technology Data Exchange (ETDEWEB)

    Gray, William J. [Lawrence Livermore National Lab. (LLNL), Livermore, CA (United States); Scannapieco, Evan [Arizona State Univ., Mesa, AZ (United States). School of Earth and Space Exploration

    2013-01-15

    Intergalactic filaments form the foundation of the cosmic web that connect galaxies together, and provide an important reservoir of gas for galaxy growth and accretion. Here we present very high resolution two-dimensional simulations of the thermal and chemical evolution of such filaments, making use of a 32 species chemistry network that tracks the evolution of key molecules formed from hydrogen, oxygen, and carbon. We study the evolution of filaments over a wide range of parameters including the initial density, initial temperature, strength of the dissociating UV background, and metallicity. In low-redshift, Z ≈ 0.1Z filaments, the evolution is determined completely by the initial cooling time. If this is sufficiently short, the center of the filament always collapses to form dense, cold core containing a substantial fraction of molecules. In high-redshift, Z = 10-3Z filaments, the collapse proceeds much more slowly. This is due mostly to the lower initial temperatures, which leads to a much more modest increase in density before the atomic cooling limit is reached, making subsequent molecular cooling much less efficient. Finally, we study how the gravitational potential from a nearby dwarf galaxy affects the collapse of the filament and compare this to NGC 5253, a nearby starbusting dwarf galaxy thought to be fueled by the accretion of filament gas. In contrast to our fiducial case, a substantial density peak forms at the center of the potential. This peak evolves faster than the rest of the filament due to the increased rate at which chemical species form and cooling occur. We find that we achieve similar accretion rates as NGC 5253, but our two-dimensional simulations do not recover the formation of the giant molecular clouds that are seen in radio observations.

  2. Current filamentation in high-current diodes

    International Nuclear Information System (INIS)

    Gordeev, A.V.; Kuksov, P.V.; Fanchenko, S.D.; Shuvaev, V.Y.

    1988-01-01

    Experimental data are reported on the filamentation of a high-current relativistic electron beam in the Kal'mar-1 relativistic-electron-beam source. A possible mechanism for this filamentation is studied theoretically. It is shown that the experimental results on the number of filaments into which the relativistic electron beam breaks up can be explained on the basis of an azimuthal nonuniformity of the current. This nonuniformity develops in the plasma near the cathode as the result of a Rayleigh--Taylor electron instability

  3. Membrane Buckling Induced by Curved Filaments

    Science.gov (United States)

    Lenz, Martin; Crow, Daniel J. G.; Joanny, Jean-François

    2009-07-01

    We present a novel buckling instability relevant to membrane budding in eukaryotic cells. In this mechanism, curved filaments bind to a lipid bilayer without changing its intrinsic curvature. As more and more filaments adsorb, newly added ones are more and more strained, which destabilizes the flat membrane. We perform a linear stability analysis of filament-dressed membranes and find that the buckling threshold is within reasonable in vivo parameter values. We account for the formation of long tubes previously observed in cells and in purified systems. We study strongly deformed dressed membranes and their bifurcation diagram numerically. Our mechanism could be validated by a simple experiment.

  4. Programmable Active Matter: Dynamics of active filaments on patterned surfaces

    Science.gov (United States)

    Yadav, Vikrant; Todd, Daniel; Milas, Peker; Ruijgrok, Paul; Bryant, Zev; Ross, Jennifer

    Interfaces are ubiquitous in biology. For a sub-cellular component moving inside the cell, any change in its local environment across an interface whether chemical concentration, density, or any other physical variables can produce novel dynamics. Recent advances in bioengineering allow us to control motor proteins' velocities when prompted by an optical trigger. Using an optical diaphragm and a gear-shifting myosin XI construct containing a photoactive LOV domain, we can spatially pattern light to create interfaces across which speed of a gliding actin filament can differ by as much as a factor of two. We observe that when a gliding actin filament crosses an interface that has a discontinuous velocity jump, it buckles and changes its angle of orientation due to the velocity mismatch. Our preliminary data suggests that for small angels of incidence, the angle of emergence increases linearly. If we increase the angle of incidence further we observe that the angle of emergence saturates. For some actin filaments approaching the interface near-tangentially we observe total internal reflection as they fail to crossover the boundary. We have modeled our system using Cytosim software package and find excellent agreement with experimental data.

  5. Novel Actin-like Filament Structure from Clostridium tetani*

    Science.gov (United States)

    Popp, David; Narita, Akihiro; Lee, Lin Jie; Ghoshdastider, Umesh; Xue, Bo; Srinivasan, Ramanujam; Balasubramanian, Mohan K.; Tanaka, Toshitsugu; Robinson, Robert C.

    2012-01-01

    Eukaryotic F-actin is constructed from two protofilaments that gently wind around each other to form a helical polymer. Several bacterial actin-like proteins (Alps) are also known to form F-actin-like helical arrangements from two protofilaments, yet with varied helical geometries. Here, we report a unique filament architecture of Alp12 from Clostridium tetani that is constructed from four protofilaments. Through fitting of an Alp12 monomer homology model into the electron microscopy data, the filament was determined to be constructed from two antiparallel strands, each composed of two parallel protofilaments. These four protofilaments form an open helical cylinder separated by a wide cleft. The molecular interactions within single protofilaments are similar to F-actin, yet interactions between protofilaments differ from those in F-actin. The filament structure and assembly and disassembly kinetics suggest Alp12 to be a dynamically unstable force-generating motor involved in segregating the pE88 plasmid, which encodes the lethal tetanus toxin, and thus a potential target for drug design. Alp12 can be repeatedly cycled between states of polymerization and dissociation, making it a novel candidate for incorporation into fuel-propelled nanobiopolymer machines. PMID:22514279

  6. Novel actin-like filament structure from Clostridium tetani.

    Science.gov (United States)

    Popp, David; Narita, Akihiro; Lee, Lin Jie; Ghoshdastider, Umesh; Xue, Bo; Srinivasan, Ramanujam; Balasubramanian, Mohan K; Tanaka, Toshitsugu; Robinson, Robert C

    2012-06-15

    Eukaryotic F-actin is constructed from two protofilaments that gently wind around each other to form a helical polymer. Several bacterial actin-like proteins (Alps) are also known to form F-actin-like helical arrangements from two protofilaments, yet with varied helical geometries. Here, we report a unique filament architecture of Alp12 from Clostridium tetani that is constructed from four protofilaments. Through fitting of an Alp12 monomer homology model into the electron microscopy data, the filament was determined to be constructed from two antiparallel strands, each composed of two parallel protofilaments. These four protofilaments form an open helical cylinder separated by a wide cleft. The molecular interactions within single protofilaments are similar to F-actin, yet interactions between protofilaments differ from those in F-actin. The filament structure and assembly and disassembly kinetics suggest Alp12 to be a dynamically unstable force-generating motor involved in segregating the pE88 plasmid, which encodes the lethal tetanus toxin, and thus a potential target for drug design. Alp12 can be repeatedly cycled between states of polymerization and dissociation, making it a novel candidate for incorporation into fuel-propelled nanobiopolymer machines.

  7. Intermediate algebra a textworkbook

    CERN Document Server

    McKeague, Charles P

    1985-01-01

    Intermediate Algebra: A Text/Workbook, Second Edition focuses on the principles, operations, and approaches involved in intermediate algebra. The publication first takes a look at basic properties and definitions, first-degree equations and inequalities, and exponents and polynomials. Discussions focus on properties of exponents, polynomials, sums, and differences, multiplication of polynomials, inequalities involving absolute value, word problems, first-degree inequalities, real numbers, opposites, reciprocals, and absolute value, and addition and subtraction of real numbers. The text then ex

  8. Intermediate algebra & analytic geometry

    CERN Document Server

    Gondin, William R

    1967-01-01

    Intermediate Algebra & Analytic Geometry Made Simple focuses on the principles, processes, calculations, and methodologies involved in intermediate algebra and analytic geometry. The publication first offers information on linear equations in two unknowns and variables, functions, and graphs. Discussions focus on graphic interpretations, explicit and implicit functions, first quadrant graphs, variables and functions, determinate and indeterminate systems, independent and dependent equations, and defective and redundant systems. The text then examines quadratic equations in one variable, system

  9. Two-step solar filament eruptions

    Science.gov (United States)

    Filippov, B.

    2018-04-01

    Coronal mass ejections (CMEs) are closely related to eruptive filaments and usually are the continuation of the same eruptive process into the upper corona. There are failed filament eruptions when a filament decelerates and stops at some greater height in the corona. Sometimes the filament after several hours starts to rise again and develops into the successful eruption with a CME formation. We propose a simple model for the interpretation of such two-step eruptions in terms of equilibrium of a flux rope in a two-scale ambient magnetic field. The eruption is caused by a slow decrease of the holding magnetic field. The presence of two critical heights for the initiation of the flux-rope vertical instability allows the flux rope to stay after the first jump some time in a metastable equilibrium near the second critical height. If the decrease of the ambient field continues, the next eruption step follows.

  10. Intense EM filamentation in relativistic hot plasmas

    Energy Technology Data Exchange (ETDEWEB)

    Hu, Qiang-Lin [Department of Physics, Jinggangshan University, Ji' an, Jiangxi 343009 (China); Chen, Zhong-Ping [Department of Physics and Institute for Fusion Studies, The University of Texas at Austin, Austin, TX 78712 (United States); Mahajan, Swadesh M., E-mail: mahajan@mail.utexas.edu [Department of Physics and Institute for Fusion Studies, The University of Texas at Austin, Austin, TX 78712 (United States); Department of Physics, School of Natural Sciences, Shiv Nadar University, Uttar Pradesh 201314 (India)

    2017-03-03

    Highlights: • Breaking up of an intense EM pulse into filaments is a spectacular demonstration of the nonlinear wave-plasma interaction. • Filaments are spectacularly sharper, highly extended and longer lived at relativistic temperatures. • EM energy concentration can trigger new nonlinear phenomena with absolute consequences for high energy density matter. - Abstract: Through 2D particle-in-cell (PIC) simulations, we demonstrate that the nature of filamentation of a high intensity electromagnetic (EM) pulse propagating in an underdense plasma, is profoundly affected at relativistically high temperatures. The “relativistic” filaments are sharper, are dramatically extended (along the direction of propagation), and live much longer than their lower temperature counterparts. The thermally boosted electron inertia is invoked to understand this very interesting and powerful phenomenon.

  11. Morgellons disease: a filamentous borrelial dermatitis

    OpenAIRE

    Middelveen MJ; Stricker RB

    2016-01-01

    Marianne J Middelveen, Raphael B Stricker International Lyme and Associated Diseases Society, Bethesda, MD, USA Abstract: Morgellons disease (MD) is a dermopathy characterized by multicolored filaments that lie under, are embedded in, or project from skin. Although MD was initially considered to be a delusional disorder, recent studies have demonstrated that the dermopathy is associated with tickborne infection, that the filaments are composed of keratin and collagen, and that they resu...

  12. Can we determine the filament chirality by the filament footpoint location or the barb-bearing?

    Science.gov (United States)

    Hao, Qi; Guo, Yang; Fang, Cheng; Chen, Peng-Fei; Cao, Wen-Da

    2016-01-01

    We attempt to propose a method for automatically detecting the solar filament chirality and barb bearing. We first introduce the concept of an unweighted undirected graph and adopt the Dijkstra shortest path algorithm to recognize the filament spine. Then, we use the polarity inversion line (PIL) shift method for measuring the polarities on both sides of the filament, and employ the connected components labeling method to identify the barbs and calculate the angle between each barb and the spine to determine the bearing of the barbs, i.e., left or right. We test the automatic detection method with Hα filtergrams from the Big Bear Solar Observatory (BBSO) Hα archive and magnetograms observed with the Helioseismic and Magnetic Imager (HMI) on board the Solar Dynamics Observatory (SDO). Four filaments are automatically detected and illustrated to show the results. The barbs in different parts of a filament may have opposite bearings. The filaments in the southern hemisphere (northern hemisphere) mainly have left-bearing (right-bearing) barbs and positive (negative) magnetic helicity, respectively. The tested results demonstrate that our method is efficient and effective in detecting the bearing of filament barbs. It is demonstrated that the conventionally believed one-to-one correspondence between filament chirality and barb bearing is not valid. The correct detection of the filament axis chirality should be done by combining both imaging morphology and magnetic field observations.

  13. Can we determine the filament chirality by the filament footpoint location or the barb-bearing?

    International Nuclear Information System (INIS)

    Hao, Qi; Guo, Yang; Fang, Cheng; Chen, Peng-Fei; Cao, Wen-Da

    2016-01-01

    We attempt to propose a method for automatically detecting the solar filament chirality and barb bearing. We first introduce the concept of an unweighted undirected graph and adopt the Dijkstra shortest path algorithm to recognize the filament spine. Then, we use the polarity inversion line (PIL) shift method for measuring the polarities on both sides of the filament, and employ the connected components labeling method to identify the barbs and calculate the angle between each barb and the spine to determine the bearing of the barbs, i.e., left or right. We test the automatic detection method with Hα filtergrams from the Big Bear Solar Observatory (BBSO) Hα archive and magnetograms observed with the Helioseismic and Magnetic Imager (HMI) on board the Solar Dynamics Observatory (SDO). Four filaments are automatically detected and illustrated to show the results. The barbs in different parts of a filament may have opposite bearings. The filaments in the southern hemisphere (northern hemisphere) mainly have left-bearing (right-bearing) barbs and positive (negative) magnetic helicity, respectively. The tested results demonstrate that our method is efficient and effective in detecting the bearing of filament barbs. It is demonstrated that the conventionally believed one-to-one correspondence between filament chirality and barb bearing is not valid. The correct detection of the filament axis chirality should be done by combining both imaging morphology and magnetic field observations. (paper)

  14. A Polymerization-Associated Structural Switch in FtsZ That Enables Treadmilling of Model Filaments

    Directory of Open Access Journals (Sweden)

    James M. Wagstaff

    2017-05-01

    Full Text Available Bacterial cell division in many organisms involves a constricting cytokinetic ring that is orchestrated by the tubulin-like protein FtsZ. FtsZ forms dynamic filaments close to the membrane at the site of division that have recently been shown to treadmill around the division ring, guiding septal wall synthesis. Here, using X-ray crystallography of Staphylococcus aureus FtsZ (SaFtsZ, we reveal how an FtsZ can adopt two functionally distinct conformations, open and closed. The open form is found in SaFtsZ filaments formed in crystals and also in soluble filaments of Escherichia coli FtsZ as deduced by electron cryomicroscopy. The closed form is found within several crystal forms of two nonpolymerizing SaFtsZ mutants and corresponds to many previous FtsZ structures from other organisms. We argue that FtsZ’s conformational switch is polymerization-associated, driven by the formation of the longitudinal intersubunit interfaces along the filament. We show that such a switch provides explanations for both how treadmilling may occur within a single-stranded filament and why filament assembly is cooperative.

  15. DAAM is required for thin filament formation and Sarcomerogenesis during muscle development in Drosophila.

    Directory of Open Access Journals (Sweden)

    Imre Molnár

    2014-02-01

    Full Text Available During muscle development, myosin and actin containing filaments assemble into the highly organized sarcomeric structure critical for muscle function. Although sarcomerogenesis clearly involves the de novo formation of actin filaments, this process remained poorly understood. Here we show that mouse and Drosophila members of the DAAM formin family are sarcomere-associated actin assembly factors enriched at the Z-disc and M-band. Analysis of dDAAM mutants revealed a pivotal role in myofibrillogenesis of larval somatic muscles, indirect flight muscles and the heart. We found that loss of dDAAM function results in multiple defects in sarcomere development including thin and thick filament disorganization, Z-disc and M-band formation, and a near complete absence of the myofibrillar lattice. Collectively, our data suggest that dDAAM is required for the initial assembly of thin filaments, and subsequently it promotes filament elongation by assembling short actin polymers that anneal to the pointed end of the growing filaments, and by antagonizing the capping protein Tropomodulin.

  16. Dynamics of contracting surfactant-covered filaments

    Science.gov (United States)

    Kamat, Pritish; Thete, Sumeet; Xu, Qi; Basaran, Osman

    2013-11-01

    When drops are produced from a nozzle, a thin liquid thread connects the primary drop that is about to form to the rest of the liquid in the nozzle. Often, the thread becomes disconnected from both the primary drop and the remnant liquid mass hanging from the nozzle and thereby gives rise to a free filament. Due to surface tension, the free filament then contracts or recoils. During recoil, the filament can either contract into a single satellite droplet or break up into several small satellites. Such satellite droplets are undesirable in applications where they can, for example, cause misting in a manufacturing environment and mar product quality in ink-jet printing. In many applications, the filaments are coated with a monolayer of surfactant. In this work, we study the dynamics of contraction of slender filaments of a Newtonian fluid that are covered with a monolayer of surfactant when the surrounding fluid is a passive gas. Taking advantage of the fact that the filaments are long and slender, we use a 1D-slender-jet approximation of the governing system of equations consisting of the Navier-Stokes system and the convection-diffusion equation for surfactant transport. We solve the 1D system of equations by a finite element based numerical method.

  17. Evidence for Mixed Helicity in Erupting Filaments

    Science.gov (United States)

    Muglach, K.; Wang, Y.-M.; Kliem, B.

    2009-09-01

    Erupting filaments are sometimes observed to undergo a rotation about the vertical direction as they rise. This rotation of the filament axis is generally interpreted as a conversion of twist into writhe in a kink-unstable magnetic flux rope. Consistent with this interpretation, the rotation is usually found to be clockwise (as viewed from above) if the post-eruption arcade has right-handed helicity, but counterclockwise if it has left-handed helicity. Here, we describe two non-active-region filament events recorded with the Extreme-Ultraviolet Imaging Telescope on the Solar and Heliospheric Observatory in which the sense of rotation appears to be opposite to that expected from the helicity of the post-event arcade. Based on these observations, we suggest that the rotation of the filament axis is, in general, determined by the net helicity of the erupting system, and that the axially aligned core of the filament can have the opposite helicity sign to the surrounding field. In most cases, the surrounding field provides the main contribution to the net helicity. In the events reported here, however, the helicity associated with the filament "barbs" is opposite in sign to and dominates that of the overlying arcade.

  18. A comparative study on the disintegration of filamentous fungi.

    Science.gov (United States)

    Taubert, J; Krings, U; Berger, R G

    2000-11-01

    Different methods for cell disintegration were tested for their efficacy on filamentous fungi, including percussion grinding, homogenization using an Ultra-Turrax, chemical treatment and lyophylization. The release of protein from Ganoderma applanatum and Pycnoporus cinnabarinus and the activity of cytoplasmatic glucose-6-phosphate dehydrogenase in the crude extracts were monitored to determine the efficiency of each disintegration technique used. Fungal cells proved to be particularly resistant towards some disintegration methods commonly used for yeasts and bacteria. Best results were obtained using a percussion grinder, if necessary, in combination with an Ultra-Turrax pretreatment.

  19. Multiple large filament bundles observed in Caulobacter crescentus by electron cryotomography

    DEFF Research Database (Denmark)

    Briegel, A; Dias, DP; Li, Z

    2006-01-01

    While the absence of any cytoskeleton was once recognized as a distinguishing feature of prokaryotes, it is now clear that a number of different bacterial proteins do form filaments in vivo. Despite the critical roles these proteins play in cell shape, genome segregation and cell division......, molecular mechanisms have remained obscure in part for lack of electron microscopy-resolution images where these filaments can be seen acting within their cellular context. Here, electron cryotomography was used to image the widely studied model prokaryote Caulobacter crescentus in an intact, near......-native state, producing three-dimensional reconstructions of these cells with unprecedented clarity and fidelity. We observed many instances of large filament bundles in various locations throughout the cell and at different stages of the cell cycle. The bundles appear to fall into four major classes based...

  20. Molecular biology of demosponge axial filaments and their roles in biosilicification.

    Science.gov (United States)

    Weaver, James C; Morse, Daniel E

    2003-11-01

    For hundreds of years, the skeletal elements of marine and freshwater sponges have intrigued investigators with a diverse array of remarkably complex morphologies. Early studies of demosponge monaxonal megascleres revealed the presence of a central organic axial filament running their entire length. Until recently, however, the precise function of these axial filaments was largely unknown. The spicules from the temperate Eastern Pacific demosponge, Tethya aurantia, comprise approximately 75% of the dry weight of this species, facilitating the large-scale isolation and purification of the biosilica-associated proteins. Silicateins, the most abundant proteins comprising the axial filaments of these spicules, prove to be members of a well-known superfamily of proteolytic and hydrolytic enzymes and can be easily collected after silica demineralization with hydrofluoric acid. Consistent with these findings, the intact filaments are more than simple, passive templates; in vitro, they actively catalyze and spatially direct the hydrolysis and polycondensation of silicon alkoxides to yield silica at neutral pH and low temperature. Catalytic activity also is exhibited by the monomeric subunits obtained by disaggregation of the protein filaments and those produced from recombinant DNA templates cloned in bacteria. These proteins also can be used to direct the polymerization of organosilicon polymers (silicones) from the corresponding organically functionalized silicon alkoxides. Based on these observations, the silicateins are currently being used as models for the design of biomimetic agents with unique catalytic and structure-directing properties. The presence of axial filaments in a diversity of spicule types and the evolutionary implications of these findings are also discussed. Copyright 2003 Wiley-Liss, Inc.

  1. Transition of Femtosecond-Filament-Solid Interactions from Single to Multiple Filament Regime.

    Science.gov (United States)

    Skrodzki, P J; Burger, M; Jovanovic, I

    2017-10-06

    High-peak-power fs-laser filaments offer unique characteristics attractive to remote sensing via techniques such as remote laser-induced breakdown spectroscopy (R-LIBS). The dynamics of several ablation mechanisms following the interaction between a filament and a solid determines the emission strength and reproducibility of target plasma, which is of relevance for R-LIBS applications. We investigate the space- and time-resolved dynamics of ionic and atomic emission from copper as well as the surrounding atmosphere in order to understand limitations of fs-filament-ablation for standoff energy delivery. Furthermore, we probe the shock front produced from filament-target interaction using time-resolved shadowgraphy and infer laser-material coupling efficiencies for both single and multiple filament regimes through analysis of shock expansion with the Sedov model for point detonation. The results provide insight into plasma structure for the range of peak powers up to 30 times the critical power for filamentation P cr . Despite the stochastic nucleation of multiple filaments at peak-powers greater than 16 P cr , emission of ionic and neutral species increases with pump beam intensity, and short-lived nitrogen emission originating from the ambient is consistently observed. Ultimately, results suggest favorable scaling of emission intensity from target species on the laser pump energy, furthering the prospects for use of filament-solid interactions for remote sensing.

  2. Role of multiple filaments in self-accelerating actions of laser filamentation in air

    Science.gov (United States)

    Hu, Yuze; Nie, Jinsong; Sun, Ke

    2017-11-01

    The nonlinear dynamics of multiple filaments in self-accelerating actions by using corrected accelerating parabolic beams (CAPBs) are numerically investigated. By increasing the number of main lobes, the curved filaments can be elongated, leading to a longer displacement. The replenished energy originating from curved multiple filaments (MFs) that constructively interfere with the central one plays a crucial role in the phenomenon. At the bifurcation position, a beam pattern in which secondary lobes tightly follow the main lobes is formed, which is beneficial for the accelerating action of MFs. A new curved filament is generated due to the merging of side-curved MFs, and its accelerating strength decreases gradually with further propagation. Moreover, a special spatiotemporal profile that enhances the possibility of acceleration is also formed. The use of the accelerating beam with the appropriate amount of main lobes provides a new approach to elongate curved filaments.

  3. Mobile communication and intermediality

    DEFF Research Database (Denmark)

    Helles, Rasmus

    2013-01-01

    The article argues the importance of intermediality as a concept for research in mobile communication and media. The constant availability of several, partially overlapping channels for communication (texting, calls, email, Facebook, etc.) requires that we adopt an integrated view of the various...

  4. an intermediate moisture meat

    African Journals Online (AJOL)

    Bunmi

    Matured leaves of Ocimum gratissimum were harvested and the extracts used to cure. Suya (an intermediate moisture meat). O. gratissimum leaves were collected from. Oyo state south west region of Nigeria, rinsed in distilled water and squeezed to extract the fluid. The meat used was Semi membranosus muscle from beef ...

  5. The significance of peroxisomes in secondary metabolite biosynthesis in filamentous fungi

    NARCIS (Netherlands)

    Bartoszewska, Magdalena; Opalinski, Lukasz; Veenhuis, Marten; van der Klei, Ida J.

    2011-01-01

    Peroxisomes are ubiquitous organelles characterized by a protein-rich matrix surrounded by a single membrane. In filamentous fungi, peroxisomes are crucial for the primary metabolism of several unusual carbon sources used for growth (e. g. fatty acids), but increasing evidence is presented that

  6. Characterization of pathway engineered strains of filamentous fungi in submerged cultures

    NARCIS (Netherlands)

    Poulsen, B.R.

    2005-01-01

    Filamentous fungi are used by the industry for making a variety of products, e.g. penicillin, high-protein food, washing powder and soft drinks. Synthesis of many of these products requires reducing equivalents in the form of NADPH, which is a potential limiting factor. The pentose phosphate pathway

  7. Addition of electrophilic lipids to actin alters filament structure

    International Nuclear Information System (INIS)

    Gayarre, Javier; Sanchez, David; Sanchez-Gomez, Francisco J.; Terron, Maria C.; Llorca, Oscar; Perez-Sala, Dolores

    2006-01-01

    Pathophysiological processes associated with oxidative stress lead to the generation of reactive lipid species. Among them, lipids bearing unsaturated aldehyde or ketone moieties can form covalent adducts with cysteine residues and modulate protein function. Through proteomic techniques we have identified actin as a target for the addition of biotinylated analogs of the cyclopentenone prostaglandins 15-deoxy-Δ 12,14 -PGJ 2 (15d-PGJ 2 ) and PGA 1 in NIH-3T3 fibroblasts. This modification could take place in vitro and mapped to the protein C-terminal end. Other electrophilic lipids, like the isoprostane 8-iso-PGA 1 and 4-hydroxy-2-nonenal, also bound to actin. The C-terminal region of actin is important for monomer-monomer interactions and polymerization. Electron microscopy showed that actin treated with 15d-PGJ 2 or 4-hydroxy-2-nonenal formed filaments which were less abundant and displayed shorter length and altered structure. Streptavidin-gold staining allowed mapping of biotinylated 15d-PGJ 2 at sites of filament disruption. These results shed light on the structural implications of actin modification by lipid electrophiles

  8. Heterologous expression of cellobiohydrolases in filamentous fungi

    DEFF Research Database (Denmark)

    Zoglowek, Marta; Lübeck, Peter S.; Ahring, Birgitte K.

    2015-01-01

    Cellobiohydrolases are among the most important enzymes functioning in the hydrolysis of crystalline cellulose, significantly contributing to the efficient biorefining of recalcitrant lignocellulosic biomass into biofuels and bio-based products. Filamentous fungi are recognized as both well...... into valuable products. However, due to low cellobiohydrolase activities, certain fungi might be deficient with regard to enzymes of value for cellulose conversion, and improving cellobiohydrolase expression in filamentous fungi has proven to be challenging. In this review, we examine the effects of altering...... promoters, signal peptides, culture conditions and host post-translational modifications. For heterologous cellobiohydrolase production in filamentous fungi to become an industrially feasible process, the construction of site-integrating plasmids, development of protease-deficient strains and glycosylation...

  9. Rosuvastatin for Primary Prevention Among Individuals With Elevated High-Sensitivity C-Reactive Protein and 5% to 10% and 10% to 20% 10-Year Risk Implications of the Justification for Use of Statins in Prevention: An Intervention Trial Evaluating Rosuvastatin (JUPITER) Trial for "Intermediate Risk"

    NARCIS (Netherlands)

    Ridker, Paul M.; Macfadyen, Jean G.; Nordestgaard, Børge G.; Koenig, Wolfgang; Kastelein, John J. P.; Genest, Jacques; Glynn, Robert J.

    2010-01-01

    Background-Recent primary prevention guidelines issued in Canada endorse the use of statin therapy among individuals at "intermediate risk" who have elevated levels of high-sensitivity C-reactive protein (hsCRP). However, trial data directly addressing whether this recommendation defines a patient

  10. Interaction of Two Filaments in a Long Filament Channel Associated with Twin Coronal Mass Ejections

    Energy Technology Data Exchange (ETDEWEB)

    Zheng, Ruisheng; Chen, Yao; Wang, Bing; Du, Guohui; Li, Chuanyang [Shandong Provincial Key Laboratory of Optical Astronomy and Solar-Terrestrial Environment, and Institute of Space Sciences, Shandong University, 264209 Weihai (China); Zhang, Qingmin [Key Laboratory for Dark Matter and Space Science, Purple Mountain Observatory, CAS, Nanjing 210008 (China); Yang, Kai, E-mail: ruishengzheng@sdu.edu.cn [School of Astronomy and Space Science, Nanjing University, 210023 Nanjing (China)

    2017-02-20

    Using the high-quality observations of the Solar Dynamics Observatory , we present the interaction of two filaments (F1 and F2) in a long filament channel associated with twin coronal mass ejections (CMEs) on 2016 January 26. Before the eruption, a sequence of rapid cancellation and emergence of the magnetic flux has been observed, which likely triggered the ascending of the west filament (F1). The east footpoints of rising F1 moved toward the east far end of the filament channel, accompanied by post-eruption loops and flare ribbons. This likely indicated a large-scale eruption involving the long filament channel, which resulted from the interaction between F1 and the east filament (F2). Some bright plasma flew over F2, and F2 stayed at rest during the eruption, likely due to the confinement of its overlying lower magnetic field. Interestingly, the impulsive F1 pushed its overlying magnetic arcades to form the first CME, and F1 finally evolved into the second CME after the collision with the nearby coronal hole. We suggest that the interaction of F1 and the overlying magnetic field of F2 led to the merging reconnection that forms a longer eruptive filament loop. Our results also provide a possible picture of the origin of twin CMEs and show that the large-scale magnetic topology of the coronal hole is important for the eventual propagation direction of CMEs.

  11. The Intermediate Neutrino Program

    Energy Technology Data Exchange (ETDEWEB)

    Adams, C.; et al.

    2015-03-23

    The US neutrino community gathered at the Workshop on the Intermediate Neutrino Program (WINP) at Brookhaven National Laboratory February 4-6, 2015 to explore opportunities in neutrino physics over the next five to ten years. Scientists from particle, astroparticle and nuclear physics participated in the workshop. The workshop examined promising opportunities for neutrino physics in the intermediate term, including possible new small to mid-scale experiments, US contributions to large experiments, upgrades to existing experiments, R&D plans and theory. The workshop was organized into two sets of parallel working group sessions, divided by physics topics and technology. Physics working groups covered topics on Sterile Neutrinos, Neutrino Mixing, Neutrino Interactions, Neutrino Properties and Astrophysical Neutrinos. Technology sessions were organized into Theory, Short-Baseline Accelerator Neutrinos, Reactor Neutrinos, Detector R&D and Source, Cyclotron and Meson Decay at Rest sessions.This report summarizes discussion and conclusions from the workshop.

  12. The Intermediate Neutrino Program

    CERN Document Server

    Adams, C.; Ankowski, A.M.; Asaadi, J.A.; Ashenfelter, J.; Axani, S.N.; Babu, K.; Backhouse, C.; Band, H.R.; Barbeau, P.S.; Barros, N.; Bernstein, A.; Betancourt, M.; Bishai, M.; Blucher, E.; Bouffard, J.; Bowden, N.; Brice, S.; Bryan, C.; Camilleri, L.; Cao, J.; Carlson, J.; Carr, R.E.; Chatterjee, A.; Chen, M.; Chen, S.; Chiu, M.; Church, E.D.; Collar, J.I.; Collin, G.; Conrad, J.M.; Convery, M.R.; Cooper, R.L.; Cowen, D.; Davoudiasl, H.; de Gouvea, A.; Dean, D.J.; Deichert, G.; Descamps, F.; DeYoung, T.; Diwan, M.V.; Djurcic, Z.; Dolinski, M.J.; Dolph, J.; Donnelly, B.; Dwyer, D.A.; Dytman, S.; Efremenko, Y.; Everett, L.L.; Fava, A.; Figueroa-Feliciano, E.; Fleming, B.; Friedland, A.; Fujikawa, B.K.; Gaisser, T.K.; Galeazzi, M.; Galehouse, D.C.; Galindo-Uribarri, A.; Garvey, G.T.; Gautam, S.; Gilje, K.E.; Gonzalez-Garcia, M.; Goodman, M.C.; Gordon, H.; Gramellini, E.; Green, M.P.; Guglielmi, A.; Hackenburg, R.W.; Hackenburg, A.; Halzen, F.; Han, K.; Hans, S.; Harris, D.; Heeger, K.M.; Herman, M.; Hill, R.; Holin, A.; Huber, P.; Jaffe, D.E.; Johnson, R.A.; Joshi, J.; Karagiorgi, G.; Kaufman, L.J.; Kayser, B.; Kettell, S.H.; Kirby, B.J.; Klein, J.R.; Kolomensky, Yu. G.; Kriske, R.M.; Lane, C.E.; Langford, T.J.; Lankford, A.; Lau, K.; Learned, J.G.; Ling, J.; Link, J.M.; Lissauer, D.; Littenberg, L.; Littlejohn, B.R.; Lockwitz, S.; Lokajicek, M.; Louis, W.C.; Luk, K.; Lykken, J.; Marciano, W.J.; Maricic, J.; Markoff, D.M.; Martinez Caicedo, D.A.; Mauger, C.; Mavrokoridis, K.; McCluskey, E.; McKeen, D.; McKeown, R.; Mills, G.; Mocioiu, I.; Monreal, B.; Mooney, M.R.; Morfin, J.G.; Mumm, P.; Napolitano, J.; Neilson, R.; Nelson, J.K.; Nessi, M.; Norcini, D.; Nova, F.; Nygren, D.R.; Orebi Gann, G.D.; Palamara, O.; Parsa, Z.; Patterson, R.; Paul, P.; Pocar, A.; Qian, X.; Raaf, J.L.; Rameika, R.; Ranucci, G.; Ray, H.; Reyna, D.; Rich, G.C.; Rodrigues, P.; Romero, E.Romero; Rosero, R.; Rountree, S.D.; Rybolt, B.; Sanchez, M.C.; Santucci, G.; Schmitz, D.; Scholberg, K.; Seckel, D.; Shaevitz, M.; Shrock, R.; Smy, M.B.; Soderberg, M.; Sonzogni, A.; Sousa, A.B.; Spitz, J.; St. John, J.M.; Stewart, J.; Strait, J.B.; Sullivan, G.; Svoboda, R.; Szelc, A.M.; Tayloe, R.; Thomson, M.A.; Toups, M.; Vacheret, A.; Vagins, M.; Van de Water, R.G.; Vogelaar, R.B.; Weber, M.; Weng, W.; Wetstein, M.; White, C.; White, B.R.; Whitehead, L.; Whittington, D.W.; Wilking, M.J.; Wilson, R.J.; Wilson, P.; Winklehner, D.; Winn, D.R.; Worcester, E.; Yang, L.; Yeh, M.; Yokley, Z.W.; Yoo, J.; Yu, B.; Yu, J.; Zhang, C.

    2015-01-01

    The US neutrino community gathered at the Workshop on the Intermediate Neutrino Program (WINP) at Brookhaven National Laboratory February 4-6, 2015 to explore opportunities in neutrino physics over the next five to ten years. Scientists from particle, astroparticle and nuclear physics participated in the workshop. The workshop examined promising opportunities for neutrino physics in the intermediate term, including possible new small to mid-scale experiments, US contributions to large experiments, upgrades to existing experiments, R&D plans and theory. The workshop was organized into two sets of parallel working group sessions, divided by physics topics and technology. Physics working groups covered topics on Sterile Neutrinos, Neutrino Mixing, Neutrino Interactions, Neutrino Properties and Astrophysical Neutrinos. Technology sessions were organized into Theory, Short-Baseline Accelerator Neutrinos, Reactor Neutrinos, Detector R&D and Source, Cyclotron and Meson Decay at Rest sessions.This report summ...

  13. The Intermediate Neutrino Program

    Energy Technology Data Exchange (ETDEWEB)

    Adams, C. [Yale Univ., New Haven, CT (United States); Alonso, J. R. [Massachusetts Inst. of Technology (MIT), Cambridge, MA (United States); Ankowski, A. M. [Virginia Polytechnic Inst. and State Univ. (Virginia Tech), Blacksburg, VA (United States); Asaadi, J. A. [Syracuse Univ., NY (United States); Ashenfelter, J. [Yale Univ., New Haven, CT (United States); Axani, S. N. [Massachusetts Inst. of Technology (MIT), Cambridge, MA (United States); Babu, K [Oklahoma State Univ., Stillwater, OK (United States); Backhouse, C. [California Inst. of Technology (CalTech), Pasadena, CA (United States); Band, H. R. [Yale Univ., New Haven, CT (United States); Barbeau, P. S. [Duke Univ., Durham, NC (United States); Barros, N. [Univ. of Pennsylvania, Philadelphia, PA (United States); Bernstein, A. [Lawrence Livermore National Lab. (LLNL), Livermore, CA (United States); Betancourt, M. [Illinois Inst. of Technology, Chicago, IL (United States); Bishai, M. [Brookhaven National Lab. (BNL), Upton, NY (United States); Blucher, E. [Univ. of Chicago, IL (United States); Bouffard, J. [State Univ. of New York (SUNY), Albany, NY (United States); Bowden, N. [Lawrence Livermore National Lab. (LLNL), Livermore, CA (United States); Brice, S. [Illinois Inst. of Technology, Chicago, IL (United States); Bryan, C. [Oak Ridge National Lab. (ORNL), Oak Ridge, TN (United States); Camilleri, L. [Columbia Univ., New York, NY (United States); Cao, J. [Inst. of High Energy Physics, Beijing (China); Carlson, J. [Los Alamos National Lab. (LANL), Los Alamos, NM (United States); Carr, R. E. [Columbia Univ., New York, NY (United States); Chatterjee, A. [Univ. of Texas, Arlington, TX (United States); Chen, M. [Univ. of California, Irvine, CA (United States); Chen, S. [Tsinghua Univ., Beijing (China); Chiu, M. [Brookhaven National Lab. (BNL), Upton, NY (United States); Church, E. D. [Illinois Inst. of Technology, Chicago, IL (United States); Pacific Northwest National Lab. (PNNL), Richland, WA (United States); Collar, J. I. [Univ. of Chicago, IL (United States); Collin, G. [Massachusetts Inst. of Technology (MIT), Cambridge, MA (United States); Conrad, J. M. [Massachusetts Inst. of Technology (MIT), Cambridge, MA (United States); Convery, M. R. [SLAC National Accelerator Lab., Menlo Park, CA (United States); Cooper, R. L. [Indiana Univ., Bloomington, IN (United States); Cowen, D. [Pennsylvania State Univ., University Park, PA (United States); Davoudiasl, H. [Brookhaven National Lab. (BNL), Upton, NY (United States); Gouvea, A. D. [Northwestern Univ., Evanston, IL (United States); Dean, D. J. [Oak Ridge National Lab. (ORNL), Oak Ridge, TN (United States); Deichert, G. [Oak Ridge National Lab. (ORNL), Oak Ridge, TN (United States); Descamps, F. [Lawrence Berkeley National Lab. (LBNL), Berkeley, CA (United States); DeYoung, T. [Michigan State Univ., East Lansing, MI (United States); Diwan, M. V. [Brookhaven National Lab. (BNL), Upton, NY (United States); Djurcic, Z. [Argonne National Lab. (ANL), Argonne, IL (United States); Dolinski, M. J. [Drexel Univ., Philadelphia, PA (United States); Dolph, J. [Brookhaven National Lab. (BNL), Upton, NY (United States); Donnelly, B. [Massachusetts Inst. of Technology (MIT), Cambridge, MA (United States); Dwyer, D. A. [Lawrence Berkeley National Lab. (LBNL), Berkeley, CA (United States); Dytman, S. [Univ. of Pittsburgh, PA (United States); Efremenko, Y. [Univ. of Tennessee, Knoxville, TN (United States); Everett, L. L. [Univ. of Wisconsin, Madison, WI (United States); Fava, A. [University of Padua, Padova (Italy); Figueroa-Feliciano, E. [Massachusetts Inst. of Technology (MIT), Cambridge, MA (United States); Fleming, B. [Yale Univ., New Haven, CT (United States); Friedland, A. [Los Alamos National Lab. (LANL), Los Alamos, NM (United States); Fujikawa, B. K. [Lawrence Berkeley National Lab. (LBNL), Berkeley, CA (United States); Gaisser, T. K. [Univ. of Delaware, Newark, DE (United States); Galeazzi, M. [Univ. of Miami, FL (United States); Galehouse, DC [Univ. of Akron, OH (United States); Galindo-Uribarri, A. [Oak Ridge National Lab. (ORNL), Oak Ridge, TN (United States); Garvey, G. T. [Los Alamos National Lab. (LANL), Los Alamos, NM (United States); Gautam, S. [Tribhuvan Univ., Kirtipur (Nepal); Gilje, K. E. [Illinois Inst. of Technology, Chicago, IL (United States); Gonzalez-Garcia, M. [Stony Brook Univ., NY (United States); Goodman, M. C. [Argonne National Lab. (ANL), Argonne, IL (United States); Gordon, H. [Brookhaven National Lab. (BNL), Upton, NY (United States); Gramellini, E. [Yale Univ., New Haven, CT (United States); Green, M. P. [Oak Ridge National Lab. (ORNL), Oak Ridge, TN (United States); Guglielmi, A. [University of Padua, Padova (Italy); Hackenburg, R. W. [Brookhaven National Lab. (BNL), Upton, NY (United States); Hackenburg, A. [Yale Univ., New Haven, CT (United States); Halzen, F. [Univ. of Wisconsin, Madison, WI (United States); Han, K. [Yale Univ., New Haven, CT (United States); Hans, S. [Brookhaven National Lab. (BNL), Upton, NY (United States); Harris, D. [Illinois Inst. of Technology, Chicago, IL (United States); Heeger, K. M. [Yale Univ., New Haven, CT (United States); Herman, M. [Brookhaven National Lab. (BNL), Upton, NY (United States); Hill, R. [Univ. of Chicago, IL (United States); Holin, A. [Univ. College London, Bloomsbury (United Kingdom); Huber, P. [Virginia Polytechnic Inst. and State Univ. (Virginia Tech), Blacksburg, VA (United States); Jaffe, D. E. [Brookhaven National Lab. (BNL), Upton, NY (United States); Johnson, R. A. [Univ. of Cincinnati, OH (United States); Joshi, J. [Brookhaven National Lab. (BNL), Upton, NY (United States); Karagiorgi, G. [Univ. of Manchester (United Kingdom); Kaufman, L. J. [Indiana Univ., Bloomington, IN (United States); Kayser, B. [Illinois Inst. of Technology, Chicago, IL (United States); Kettell, S. H. [Brookhaven National Lab. (BNL), Upton, NY (United States); Kirby, B. J. [Brookhaven National Lab. (BNL), Upton, NY (United States); Klein, J. R. [Univ. of Texas, Arlington, TX (United States); Kolomensky, Y. G. [Lawrence Berkeley National Lab. (LBNL), Berkeley, CA (United States); Univ. of California, Berkeley, CA (United States); Kriske, R. M. [Univ. of Minnesota, Minneapolis, MN (United States); Lane, C. E. [Drexel Univ., Philadelphia, PA (United States); Langford, T. J. [Yale Univ., New Haven, CT (United States); Lankford, A. [Univ. of California, Irvine, CA (United States); Lau, K. [Univ. of Houston, TX (United States); Learned, J. G. [Univ. of Hawaii, Honolulu, HI (United States); Ling, J. [Univ. of Illinois, Urbana-Champaign, IL (United States); Link, J. M. [Virginia Polytechnic Inst. and State Univ. (Virginia Tech), Blacksburg, VA (United States); Lissauer, D. [Brookhaven National Lab. (BNL), Upton, NY (United States); Littenberg, L. [Brookhaven National Lab. (BNL), Upton, NY (United States); Littlejohn, B. R. [Illinois Inst. of Technology, Chicago, IL (United States); Lockwitz, S. [Illinois Inst. of Technology, Chicago, IL (United States); Lokajicek, M. [Inst. of Physics of the Academy of Sciences of Czech Republic, Prague (Czech Republic); Louis, W. C. [Los Alamos National Lab. (LANL), Los Alamos, NM (United States); Luk, K. [Univ. of California, Berkeley, CA (United States); Lykken, J. [Illinois Inst. of Technology, Chicago, IL (United States); Marciano, W. J. [Brookhaven National Lab. (BNL), Upton, NY (United States); Maricic, J. [Univ. of Hawaii, Honolulu, HI (United States); Markoff, D. M. [North Carolina Central Univ., Durham, NC (United States); Caicedo, D. A. M. [Illinois Inst. of Technology, Chicago, IL (United States); Mauger, C. [Los Alamos National Lab. (LANL), Los Alamos, NM (United States); Mavrokoridis, K. [Univ. of Liverpool (United Kingdom); McCluskey, E. [Illinois Inst. of Technology, Chicago, IL (United States); McKeen, D. [Univ. of Washington, Seattle, WA (United States); McKeown, R. [Thomas Jefferson National Accelerator Facility (TJNAF), Newport News, VA (United States); Mills, G. [Los Alamos National Lab. (LANL), Los Alamos, NM (United States); Mocioiu, I. [Pennsylvania State Univ., University Park, PA (United States); Monreal, B. [Univ. of California, Santa Barbara, CA (United States); Mooney, M. R. [Brookhaven National Lab. (BNL), Upton, NY (United States); Morfin, J. G. [Illinois Inst. of Technology, Chicago, IL (United States); Mumm, P. [National Inst. of Standards and Technology (NIST), Boulder, CO (United States); Napolitano, J. [Temple Univ., Philadelphia, PA (United States); Neilson, R. [Drexel Univ., Philadelphia, PA (United States); Nelson, J. K. [College of William and Mary, Williamsburg, VA (United States); Nessi, M. [European Organization for Nuclear Research (CERN), Geneva (Switzerland); Norcini, D. [Yale Univ., New Haven, CT (United States); Nova, F. [Univ. of Texas, Austin, TX (United States); Nygren, D. R. [Univ. of Texas, Arlington, TX (United States); Gann, GDO [Lawrence Berkeley National Lab. (LBNL), Berkeley, CA (United States); Univ. of California, Berkeley, CA (United States); Palamara, O. [Illinois Inst. of Technology, Chicago, IL (United States); Parsa, Z. [Brookhaven National Lab. (BNL), Upton, NY (United States); Patterson, R. [California Inst. of Technology (CalTech), Pasadena, CA (United States); Paul, P. [Stony Brook Univ., NY (United States); Pocar, A. [Univ. of Massachusetts, Amherst, MA (United States); Qian, X. [Brookhaven National Lab. (BNL), Upton, NY (United States); Raaf, J. L. [Illinois Inst. of Technology, Chicago, IL (United States); Rameika, R. [Illinois Inst. of Technology, Chicago, IL (United States); Ranucci, G. [National Inst. of Nuclear Physics, Milano (Italy); Ray, H. [Univ. of Florida, Gainesville, FL (United States); Reyna, D. [Sandia National Lab. (SNL-NM), Albuquerque, NM (United States); Rich, G. C. [Triangle Universities Nuclear Lab., Durham, NC (United States); Rodrigues, P. [Univ. of Rochester, NY (United States); Romero, E. R. [Oak Ridge National Lab. (ORNL), Oak Ridge, TN (United States); Univ. of Tennessee, Knoxville, TN (United States); Rosero, R. [Brookhaven National Lab. (BNL), Upton, NY (United States); Rountree, S. D. [Virginia Polytechnic Inst. and State Univ. (Virginia Tech), Blacksburg, VA (United States); Rybolt, B. [Univ. of Tennessee, Knoxville, TN (United States); Sanchez, M. C. [Iowa State Univ., Ames, IA (United States); Santucci, G. [Stony Brook Univ., NY (United States); Schmitz, D. [Univ. of Chicago, IL (United States); Scholberg, K. [Duke Univ., Durham, NC (United States); Seckel, D. [Univ. of Delaware, Newark, DE (United States); Shaevitz, M. [Columbia Univ., New York, NY (United States); Shrock, R. [Stony Brook Univ., NY (United States); Smy, M. B. [Univ. of California, Irvine, CA (United States); Soderberg, M. [Syracuse Univ., NY (United States); Sonzogni, A. [Brookhaven National Lab. (BNL), Upton, NY (United States); Sousa, A. B. [Univ. of Cincinnati, OH (United States); Spitz, J. [Massachusetts Inst. of Technology (MIT), Cambridge, MA (United States); John, J. M. S. [Univ. of Cincinnati, OH (United States); Stewart, J. [Brookhaven National Lab. (BNL), Upton, NY (United States); Strait, J. B. [Illinois Inst. of Technology, Chicago, IL (United States); Sullivan, G. [Univ. of Maryland, College Park, MD (United States); Svoboda, R. [Univ. of California, Davis, CA (United States); Szelc, A. M. [Yale Univ., New Haven, CT (United States); Tayloe, R. [Indiana Univ., Bloomington, IN (United States); Thomson, M. A. [Univ. of Cambridge (United Kingdom); Toups, M. [Massachusetts Inst. of Technology (MIT), Cambridge, MA (United States); Vacheret, A. [Univ. of Oxford (United Kingdom); Vagins, M. [Univ. of California, Irvine, CA (United States); Water, R. G. V. D. [Los Alamos National Lab. (LANL), Los Alamos, NM (United States); Vogelaar, R. B. [Virginia Polytechnic Inst. and State Univ. (Virginia Tech), Blacksburg, VA (United States); Weber, M. [Bern (Switzerland); Weng, W. [Brookhaven National Lab. (BNL), Upton, NY (United States); Wetstein, M. [Univ. of Chicago, IL (United States); White, C. [Oak Ridge National Lab. (ORNL), Oak Ridge, TN (United States); White, B. R. [Oak Ridge National Lab. (ORNL), Oak Ridge, TN (United States); Whitehead, L. [Univ. of Houston, TX (United States); Whittington, D. W. [Indiana Univ., Bloomington, IN (United States); Wilking, M. J. [Stony Brook Univ., NY (United States); Wilson, R. J. [Colorado State Univ., Fort Collins, CO (United States); Wilson, P. [Illinois Inst. of Technology, Chicago, IL (United States); Winklehner, D. [Massachusetts Inst. of Technology (MIT), Cambridge, MA (United States); Winn, D. R. [Fairfield Univ., CT (United States); Worcester, E. [Brookhaven National Lab. (BNL), Upton, NY (United States); Yang, L. [Univ. of Illinois, Urbana-Champaign, IL (United States); Yeh, M [Brookhaven National Lab. (BNL), Upton, NY (United States); Yokley, Z. W. [Virginia Polytechnic Inst. and State Univ. (Virginia Tech), Blacksburg, VA (United States); Yoo, J. [Illinois Inst. of Technology, Chicago, IL (United States); Yu, B. [Brookhaven National Lab. (BNL), Upton, NY (United States); Yu, J. [Univ. of Texas, Arlington, TX (United States); Zhang, C. [Brookhaven National Lab. (BNL), Upton, NY (United States)

    2017-04-03

    The US neutrino community gathered at the Workshop on the Intermediate Neutrino Program (WINP) at Brookhaven National Laboratory February 4-6, 2015 to explore opportunities in neutrino physics over the next five to ten years. Scientists from particle, astroparticle and nuclear physics participated in the workshop. The workshop examined promising opportunities for neutrino physics in the intermediate term, including possible new small to mid-scale experiments, US contributions to large experiments, upgrades to existing experiments, R&D plans and theory. The workshop was organized into two sets of parallel working group sessions, divided by physics topics and technology. Physics working groups covered topics on Sterile Neutrinos, Neutrino Mixing, Neutrino Interactions, Neutrino Properties and Astrophysical Neutrinos. Technology sessions were organized into Theory, Short-Baseline Accelerator Neutrinos, Reactor Neutrinos, Detector R&D and Source, Cyclotron and Meson Decay at Rest sessions.This report summarizes discussion and conclusions from the workshop.

  14. Effects of solution crowding on actin polymerization reveal the energetic basis for nucleotide-dependent filament stability.

    Science.gov (United States)

    Frederick, Kendra B; Sept, David; De La Cruz, Enrique M

    2008-05-02

    Actin polymerization is a fundamental cellular process involved in cell structure maintenance, force generation, and motility. Phosphate release from filament subunits following ATP hydrolysis destabilizes the filament lattice and increases the critical concentration (C(c)) for assembly. The structural differences between ATP- and ADP-actin are still debated, as well as the energetic factors that underlie nucleotide-dependent filament stability, particularly under crowded intracellular conditions. Here, we investigate the effect of crowding agents on ATP- and ADP-actin polymerization and find that ATP-actin polymerization is largely unaffected by solution crowding, while crowding agents lower the C(c) of ADP-actin in a concentration-dependent manner. The stabilities of ATP- and ADP-actin filaments are comparable in the presence of physiological amounts (approximately 30% w/v) and types (sorbitol) of low molecular weight crowding agents. Crowding agents act to stabilize ADP-F-actin by slowing subunit dissociation. These observations suggest that nucleotide hydrolysis and phosphate release per se do not introduce intrinsic differences in the in vivo filament stability. Rather, the preferential disassembly of ADP-actin filaments in cells is driven through interactions with regulatory proteins. Interpretation of the experimental data according to osmotic stress theory implicates water as an allosteric regulator of actin activity and hydration as the molecular basis for nucleotide-dependent filament stability.

  15. Intermediate energy data

    International Nuclear Information System (INIS)

    Koning, A.J.; Fukahori, T.; Hasegawa, A.

    1998-01-01

    Subgroup 13 (SG13) on Intermediate Energy Nuclear data was formed by NEA Nuclear Science Committee to solve common problems of these types of data for nuclear applications. An overview is presented in this final report of the present activities of SG13, including data needs, high-priority nuclear data request list (nuclides), compilation of experimental data, specialists meetings and benchmarks, data formats and data libraries. Some important accomplishments are summarized, and recommendations are presented. (R.P.)

  16. The Selective Autophagy Receptor p62 Forms a Flexible Filamentous Helical Scaffold

    Directory of Open Access Journals (Sweden)

    Rodolfo Ciuffa

    2015-05-01

    Full Text Available The scaffold protein p62/SQSTM1 is involved in protein turnover and signaling and is commonly found in dense protein bodies in eukaryotic cells. In autophagy, p62 acts as a selective autophagy receptor that recognizes and shuttles ubiquitinated proteins to the autophagosome for degradation. The structural organization of p62 in cellular bodies and the interplay of these assemblies with ubiquitin and the autophagic marker LC3 remain to be elucidated. Here, we present a cryo-EM structural analysis of p62. Together with structures of assemblies from the PB1 domain, we show that p62 is organized in flexible polymers with the PB1 domain constituting a helical scaffold. Filamentous p62 is capable of binding LC3 and addition of long ubiquitin chains induces disassembly and shortening of filaments. These studies explain how p62 assemblies provide a large molecular scaffold for the nascent autophagosome and reveal how they can bind ubiquitinated cargo.

  17. Comparative analysis of programmed cell death pathways in filamentous fungi

    Directory of Open Access Journals (Sweden)

    Wortman Jennifer R

    2005-12-01

    Full Text Available Abstract Background Fungi can undergo autophagic- or apoptotic-type programmed cell death (PCD on exposure to antifungal agents, developmental signals, and stress factors. Filamentous fungi can also exhibit a form of cell death called heterokaryon incompatibility (HI triggered by fusion between two genetically incompatible individuals. With the availability of recently sequenced genomes of Aspergillus fumigatus and several related species, we were able to define putative components of fungi-specific death pathways and the ancestral core apoptotic machinery shared by all fungi and metazoa. Results Phylogenetic profiling of HI-associated proteins from four Aspergilli and seven other fungal species revealed lineage-specific protein families, orphan genes, and core genes conserved across all fungi and metazoa. The Aspergilli-specific domain architectures include NACHT family NTPases, which may function as key integrators of stress and nutrient availability signals. They are often found fused to putative effector domains such as Pfs, SesB/LipA, and a newly identified domain, HET-s/LopB. Many putative HI inducers and mediators are specific to filamentous fungi and not found in unicellular yeasts. In addition to their role in HI, several of them appear to be involved in regulation of cell cycle, development and sexual differentiation. Finally, the Aspergilli possess many putative downstream components of the mammalian apoptotic machinery including several proteins not found in the model yeast, Saccharomyces cerevisiae. Conclusion Our analysis identified more than 100 putative PCD associated genes in the Aspergilli, which may help expand the range of currently available treatments for aspergillosis and other invasive fungal diseases. The list includes species-specific protein families as well as conserved core components of the ancestral PCD machinery shared by fungi and metazoa.

  18. High-resolution Observations of Sympathetic Filament Eruptions by NVST

    Energy Technology Data Exchange (ETDEWEB)

    Li, Shangwei; Su, Yingna; Zhou, Tuanhui; Ji, Haisheng [Key Laboratory for Dark Matter and Space Science, Purple Mountain Observatory, CAS, Nanjing 210008 (China); Van Ballegooijen, Adriaan [5001 Riverwood Avenue, Sarasota, FL 34231 (United States); Sun, Xudong, E-mail: ynsu@pmo.ac.cn [W. W. Hansen Experimental Physics Laboratory, Stanford University, Stanford, CA 94305 (United States)

    2017-07-20

    We investigate two sympathetic filament eruptions observed by the New Vacuum Solar Telescope on 2015 October 15. The full picture of the eruptions is obtained from the corresponding Solar Dynamics Observatory ( SDO )/Atmospheric Imaging Assembly (AIA) observations. The two filaments start from active region NOAA 12434 in the north and end in one large quiescent filament channel in the south. The left filament erupts first, followed by the right filament eruption about 10 minutes later. Clear twist structure and rotating motion are observed in both filaments during the eruption. Both eruptions failed, since the filaments first rise up, then flow toward the south and merge into the southern large quiescent filament. We also observe repeated activations of mini filaments below the right filament after its eruption. Using magnetic field models constructed based on SDO /HMI magnetograms via the flux rope insertion method, we find that the left filament eruption is likely to be triggered by kink instability, while the weakening of overlying magnetic fields due to magnetic reconnection at an X-point between the two filament systems might play an important role in the onset of the right filament eruption.

  19. Filament networks attached to membranes: cytoskeletal pressure and local bilayer deformation

    Energy Technology Data Exchange (ETDEWEB)

    Auth, Thorsten [Department of Materials and Interfaces, Weizmann Institute of Science, PO Box 26, Rehovot 76100 (Israel); Safran, S A [Department of Materials and Interfaces, Weizmann Institute of Science, PO Box 26, Rehovot 76100 (Israel); Gov, Nir S [Department of Chemical Physics, Weizmann Institute of Science, PO Box 26, Rehovot 76100 (Israel)

    2007-11-15

    Several cell types, among them red blood cells, have a cortical, two-dimensional (2D) network of filaments sparsely attached to their lipid bilayer. In many mammalian cells, this 2D polymer network is connected to an underlying 3D, more rigid cytoskeleton. In this paper, we consider the pressure exerted by the thermally fluctuating, cortical network of filaments on the bilayer and predict the bilayer deformations that are induced by this pressure. We treat the filaments as flexible polymers and calculate the pressure that a network of such linear chains exerts on the bilayer; we then minimize the bilayer shape in order to predict the resulting local deformations. We compare our predictions with membrane deformations observed in electron micrographs of red blood cells. The polymer pressure along with the resulting membrane deformation can lead to compartmentalization, regulate in-plane diffusion and may influence protein sorting as well as transmit signals to the polymerization of the underlying 3D cytoskeleton.

  20. Filament networks attached to membranes: cytoskeletal pressure and local bilayer deformation

    International Nuclear Information System (INIS)

    Auth, Thorsten; Safran, S A; Gov, Nir S

    2007-01-01

    Several cell types, among them red blood cells, have a cortical, two-dimensional (2D) network of filaments sparsely attached to their lipid bilayer. In many mammalian cells, this 2D polymer network is connected to an underlying 3D, more rigid cytoskeleton. In this paper, we consider the pressure exerted by the thermally fluctuating, cortical network of filaments on the bilayer and predict the bilayer deformations that are induced by this pressure. We treat the filaments as flexible polymers and calculate the pressure that a network of such linear chains exerts on the bilayer; we then minimize the bilayer shape in order to predict the resulting local deformations. We compare our predictions with membrane deformations observed in electron micrographs of red blood cells. The polymer pressure along with the resulting membrane deformation can lead to compartmentalization, regulate in-plane diffusion and may influence protein sorting as well as transmit signals to the polymerization of the underlying 3D cytoskeleton

  1. Changes in carbon footprint when integrating production of filamentous fungi in 1st generation ethanol plants.

    Science.gov (United States)

    Brancoli, Pedro; Ferreira, Jorge A; Bolton, Kim; Taherzadeh, Mohammad J

    2018-02-01

    Integrating the cultivation of edible filamentous fungi in the thin stillage from ethanol production is presently being considered. This integration can increase the ethanol yield while simultaneously producing a new value-added protein-rich biomass that can be used for animal feed. This study uses life cycle assessment to determine the change in greenhouse gas (GHG) emissions when integrating the cultivation of filamentous fungi in ethanol production. The result shows that the integration performs better than the current scenario when the fungal biomass is used as cattle feed for system expansion and when energy allocation is used. It performs worse if the biomass is used as fish feed. Hence, integrating the cultivation of filamentous fungi in 1st generation ethanol plants combined with proper use of the fungi can lead to a reduction of GHG emissions which, considering the number of existing ethanol plants, can have a significant global impact. Copyright © 2017 Elsevier Ltd. All rights reserved.

  2. Transition from embryonic to adult epidermis in reptiles occurs by the production of corneous beta-proteins.

    Science.gov (United States)

    Alibardi, Lorenzo

    2014-01-01

    The adaptation of the epidermis in amniote vertebrates to life on land took place by a drastic change from an embryonic epidermis made of two-four periderm layers to a terrestrial-proof epidermis. This transition occurred by the increase in types and number of specialized corneous proteins coded by genes of the Epidermal Differentiation Complex. The prevalent types of corneous proteins produced in the reptilian epidermis contain a beta-sheet region of high amino acid homology which allows their polymerization into a meshwork of filaments forming the hard corneous material of scales and claws. The present immunogold ultrastructural study shows that this transition occurs with the synthesis of glycine-rich corneous beta-proteins (formerly indicated as beta-keratins) that are added to the initial framework of acidic intermediate filaments produced in the embryonic epidermis of lizards, snake, alligator and turtle. These corneous beta-proteins are accumulated in the transitional and definitive layers of reptilian epidermis formed underneath the transitory two-four layered embryonic epidermis. In the more specialized reptiles capable of shedding the epidermis as a single unit, such as lizards and snakes, special glycine-cysteine rich beta-proteins are initially produced in a single layer immediately formed beneath the embryonic epidermis, the oberhautchen. The latter layer allows the in ovo shedding of the embryonic epidermis in preparation for hatching, and in the following shedding cycles of the adult epidermis. The production of specialized corneous-specific beta-proteins in addition to intermediate filament keratins was probably an essential addition for terrestrial life during the evolution of reptiles into different lineages, including birds. The increase of glycine and cysteine in epidermal proteins enhanced the hydrophobicity, insolubility and mechanical strength of the stratum corneum in these amniotes.

  3. Drug delivery vectors based on filamentous bacteriophages and phage-mimetic nanoparticles.

    Science.gov (United States)

    Ju, Zhigang; Sun, Wei

    2017-11-01

    With the development of nanomedicine, a mass of nanocarriers have been exploited and utilized for targeted drug delivery, including liposomes, polymers, nanoparticles, viruses, and stem cells. Due to huge surface bearing capacity and flexible genetic engineering property, filamentous bacteriophage and phage-mimetic nanoparticles are attracting more and more attentions. As a rod-like bio-nanofiber without tropism to mammalian cells, filamentous phage can be easily loaded with drugs and directly delivered to the lesion location. In particular, chemical drugs can be conjugated on phage surface by chemical modification, and gene drugs can also be inserted into the genome of phage by recombinant DNA technology. Meanwhile, specific peptides/proteins displayed on the phage surface are able to conjugate with nanoparticles which will endow them specific-targeting and huge drug-loading capacity. Additionally, phage peptides/proteins can directly self-assemble into phage-mimetic nanoparticles which may be applied for self-navigating drug delivery nanovehicles. In this review, we summarize the production of phage particles, the identification of targeting peptides, and the recent applications of filamentous bacteriophages as well as their protein/peptide for targeting drug delivery in vitro and in vivo. The improvement of our understanding of filamentous bacteriophage and phage-mimetic nanoparticles will supply new tools for biotechnological approaches.

  4. Evaluating biochemical response of filamentous algae integrated with different water bodies.

    Science.gov (United States)

    Çelekli, Abuzer; Kapı, Emine; Soysal, Çiğdem; Arslanargun, Hamdullah; Bozkurt, Hüseyin

    2017-08-01

    The present study prompted the second attempts to evaluate biochemical responses of filamentous algae under different physico-chemical variables in various water bodies in Turkey. These variables were investigated by use of multivariate approach in the years of 2013 (May and November) and 2014 (May and October). Studied ecoregions had the different geographic position, climate, land-use, and anthropogenic activities, could strongly affect physico-chemical variables of water bodies, which caused to change or regulate in algal biomass composition due to the different response of filamentous species. Besides, biochemical responses of species changed at different sampling times and stations. Multivariate analyses indicated that temperature, heavy metals, and nutrient contents of aquatic systems were found to be major variables driving the spatial and temporal occurrence and biochemical contents of filamentous species. Total protein and pigment production by filamentous algae were high in water bodies having high nutrients, whereas they were low in high heavy metal contents. Amount of malondialdehyde (MDA), H 2 O 2 , total thiol group, total phenolic compounds, proline, total carbohydrate, and bioaccumulation of metals by filamentous algae were closely related with heavy metal contents of water bodies, indicated by the multivariate approach. Significant increase in aforementioned biochemical compounds with a distinct range of habitats and sensitive-tolerance to environmental conditions could make them highly valuable indicators. Copyright © 2017 Elsevier Inc. All rights reserved.

  5. The length of cooperative units on the thin filament in rabbit psoas muscle fibres.

    Science.gov (United States)

    Ding, Wei; Fujita, Hideaki; Kawai, Masataka

    2002-11-01

    The length of cooperative units along the thin filament of rabbit psoas single muscle fibres was determined by reducing filament length by treatment with the thin filament severing protein, gelsolin, in the presence of Ca(2+) and 2,3-butanedione 2-monoxime (BDM). The average time for 50 % reduction in isometric tension was 6.7 min at 22 degrees C. The pCa-tension relationship was measured at 22 degrees C, pH 7.00 and ionic strength 200 mM, and the data were fitted to the Hill equation to determine the half-saturation point (K) and the cooperativity (n). Our results demonstrate that the cooperativity does not change much when the remaining isometric tension was in the range 20-100 %. The cooperativity quickly diminished when the remaining tension was reduced to less than 20 %. Our results further demonstrate that the change in the pK value was minimal and averaged 0.075 (less Ca(2+) sensitive) as the thin filament length was reduced. We infer from the first observation that the thin filament cooperativity extends up to 0.2 microm, which includes the maximum of about four basic cooperative units consisting of seven actin molecules, one tropomyosin dimer and one troponin complex. We infer from the second observation that the Ca(2+) sensitivity is slightly reduced by removal of the cooperative interaction between neighbouring cooperative units.

  6. Altered expression of epithelial cell surface glycoconjugates and intermediate filaments at the margins of mucosal wounds

    DEFF Research Database (Denmark)

    Dabelsteen, Erik; Grøn, B.; Mandel, U.

    1998-01-01

    Alterations in cell to cell adhesion are necessary to enable the type of cell movements that are associated with epithelial wound healing and malignant invasion. Several studies of transformed cells have related epithelial cell movement to changes in the cell surface expression of the carbohydrate......-T antigen. The changes induced by wounding in the expression of collagen IV, laminin gamma2-chain (laminin-5), and laminin alpha5-chain were similar to those found in skin wounds and served to define the region of epithelial movement. This region was found to show a marked increase in staining for both...... epithelium, a pattern of expression similar to K16, which was also strongly upregulated in both the outgrowth and the adjacent nonwounded epithelium. These findings provide further support for an influence of such carbohydrate structures on the migratory behavior of epithelial cells....

  7. Large-scale Motion of Solar Filaments

    Indian Academy of Sciences (India)

    tribpo

    Large-scale Motion of Solar Filaments. Pavel Ambrož, Astronomical Institute of the Acad. Sci. of the Czech Republic, CZ-25165. Ondrejov, The Czech Republic. e-mail: pambroz@asu.cas.cz. Alfred Schroll, Kanzelhöehe Solar Observatory of the University of Graz, A-9521 Treffen,. Austria. e-mail: schroll@solobskh.ac.at.

  8. Filamentous bacteria transport electrons over centimetre distances

    DEFF Research Database (Denmark)

    Pfeffer, Christian; Larsen, Steffen; Song, Jie

    2012-01-01

    across centimetre-wide zones. Here we present evidence that the native conductors are long, filamentous bacteria. They abounded in sediment zones with electric currents and along their length they contained strings with distinct properties in accordance with a function as electron transporters. Living...

  9. Modelling the morphology of filamentous microorganisms

    DEFF Research Database (Denmark)

    Nielsen, Jens Bredal

    1996-01-01

    The rapid development in image analysis techniques has made it possible to study the growth kinetics of filamentous microorganisms in more detail than previously, However, owing to the many different processes that influence the morphology it is important to apply mathematical models to extract...

  10. Filament stretching rheometry of polymer melts

    DEFF Research Database (Denmark)

    Hassager, Ole; Nielsen, Jens Kromann; Rasmussen, Henrik Koblitz

    2005-01-01

    The Filament Stretching Rheometry (FSR) method developed by Sridhar, McKinley and coworkers for polymer solutions has been extended to be used also for polymer melts. The design of a melt-FSR will be described and differences to conventional melt elongational rheometers will be pointed out. Results...

  11. The exo-metabolome in filamentous fungi

    DEFF Research Database (Denmark)

    Thrane, Ulf; Andersen, Birgitte; Frisvad, Jens Christian

    2007-01-01

    Filamentous fungi are a diverse group of eukaryotic microorganisms that have a significant impact on human life as spoilers of food and feed by degradation and toxin production. They are also most useful as a source of bulk and fine chemicals and pharmaceuticals. This chapter focuses on the exo...

  12. Evolution of genetic systems in filamentous ascomycetes

    NARCIS (Netherlands)

    Nauta, M.J.

    1994-01-01

    A great variety of genetic systems exist in filamentous ascomycetes. The transmission of genetic material does not only occur by (sexual or asexual) reproduction, but it can also follow vegetative fusion of different strains. In this thesis the evolution of this variability is studied,

  13. Featured Image: A Filament Forms and Erupts

    Science.gov (United States)

    Kohler, Susanna

    2017-06-01

    This dynamic image of active region NOAA 12241 was captured by the Solar Dynamics Observatorys Atmospheric Imaging Assembly in December 2014. Observations of this region from a number of observatories and instruments recently presented by Jincheng Wang (University of Chinese Academy of Sciences) and collaborators reveal details about the formation and eruption of a long solar filament. Wang and collaborators show that the right part of the filament formed by magnetic reconnection between two bundles of magnetic field lines, while the left part formed as a result of shearing motion. When these two parts interacted, the filament erupted. You can read more about the teams results in the article linked below. Also, check out this awesome video of the filament formation and eruption, again by SDO/AIA:http://cdn.iopscience.com/images/0004-637X/839/2/128/Full/apjaa6bf3f1_video.mp4CitationJincheng Wang et al 2017 ApJ 839 128. doi:10.3847/1538-4357/aa6bf3

  14. Large-scale Motion of Solar Filaments

    Indian Academy of Sciences (India)

    tribpo

    The 'seeing' dependent contrast of the Hα pictures is the source of uncertainties during the measurements on ... Results of measurements and conclusions. Heliographic position of the filaments is measured on the full disc Hα pictures taken ... consecutive magnetic synoptic charts. Two arrays of corresponding velocities are ...

  15. Mapping the filaments in NGC 1275

    Science.gov (United States)

    Cobos, Aracely Susan; Rich, Jeffrey; Great Observatories All-sky LIRG Survey (GOALS)

    2018-01-01

    The giant elliptical brightest cluster galaxies (BCGs) at the centers of many massive clusters are often surrounded by drawn-out forms of gaseous material. It is believed that this gaseous material is gas condensing from the intracluster medium (ICM) in a “cooling flow,” and it can directly impact the growth of the BCG. The galaxy NGC 1275 is one of the closest giant elliptical BCGs and lies at the center of the Perseus cluster. NGC 1275 has large filaments that are thought to be associated with a cooling flow, but they may also be affected by its AGN. To investigate the relationship between the AGN and the cooling flow we have mapped the filaments around NGC 1275 with the Cosmic Web Imager, an image-slicing integral field spectrograph at Palomar Observatories. We employ standard emission-line ratio diagnostics to determine the source of ionizing radiation. We use our analysis to investigate whether the formation of the extended filaments is a result of gas from the ICM collapsing onto the galaxy as it cools or if it is possible that the filaments are a result of the cluster’s interaction with the outflow driven by the AGN.

  16. On viscoelastic instability in polymeric filaments

    DEFF Research Database (Denmark)

    Rasmussen, Henrik Koblitz; Hassager, Ole

    1999-01-01

    The 3D Lagrangian Integral Method is used to simulate the effects of surface tension on the viscoelastic end-plate instability, occuring in the rapid extension of some polymeric filaments between parallel plates. It is shovn that the surface tension delays the onset of the instability. Furthermore...

  17. The keratin-binding protein Albatross regulates polarization of epithelial cells.

    Science.gov (United States)

    Sugimoto, Masahiko; Inoko, Akihito; Shiromizu, Takashi; Nakayama, Masanori; Zou, Peng; Yonemura, Shigenobu; Hayashi, Yuko; Izawa, Ichiro; Sasoh, Mikio; Uji, Yukitaka; Kaibuchi, Kozo; Kiyono, Tohru; Inagaki, Masaki

    2008-10-06

    The keratin intermediate filament network is abundant in epithelial cells, but its function in the establishment and maintenance of cell polarity is unclear. Here, we show that Albatross complexes with Par3 to regulate formation of the apical junctional complex (AJC) and maintain lateral membrane identity. In nonpolarized epithelial cells, Albatross localizes with keratin filaments, whereas in polarized epithelial cells, Albatross is primarily localized in the vicinity of the AJC. Knockdown of Albatross in polarized cells causes a disappearance of key components of the AJC at cell-cell borders and keratin filament reorganization. Lateral proteins E-cadherin and desmoglein 2 were mislocalized even on the apical side. Although Albatross promotes localization of Par3 to the AJC, Par3 and ezrin are still retained at the apical surface in Albatross knockdown cells, which retain intact microvilli. Analysis of keratin-deficient epithelial cells revealed that keratins are required to stabilize the Albatross protein, thus promoting the formation of AJC. We propose that keratins and the keratin-binding protein Albatross are important for epithelial cell polarization.

  18. Isolation and characterization of flagellar filaments from Bacillus cereus ATCC 14579.

    Science.gov (United States)

    Tagawa, Yuichi

    2014-12-01

    Isolated flagellar filaments from the type strain of Bacillus cereus, ATCC 14579, were shown to consist of 34, 32 and 31 kDa proteins in similar proportions as judged by band intensities on sodium dodecyl sulfate-polyacrylamide gel electrophoresis. The N-terminal amino acid sequences of these three proteins of strain ATCC 14579 were identical with the deduced sequences of three flagellin genes BC1657, BC1658 and BC1659 in the whole genome sequence. Strain ATCC 14579 was classified into serotype T2 by a flagellar serotyping scheme for B. cereus strains that are untypeable into known flagellar serotypes H1 to H23. Flagellar filaments from a reference strain of serotype T2 contained two protein bands at 34 and 32 kDa, but a single protein band at 39 kDa was detected in flagellar filaments of a reference strain of serotype H1. Two murine monoclonal antibodies, 1A5 and 2A5, which recognize both the 34 and 32 kDa flagellins and a single flagellin of 32 kDa, respectively, were specifically reactive with B. cereus strains ATCC 14579 and serotype T2 in whole-cell ELISA and bacterial motility inhibition tests. In immunoelectron microscopy with monoclonal antibodies 1A5 and 2A5, colloidal gold spheres were shown to localize almost evenly over the entire part of flagellar filaments. Since strain ATCC 14579, and presumably strain serotype T2, are unusual among B. cereus strains in possessing multiple genes that encode flagellin subunits, a possible unique mechanism may contribute to assembly of multiple flagellin subunits into the filament over its entire length.

  19. Harmful impact of filamentous algae (Spirogyra sp.) on juvenile crayfish

    OpenAIRE

    Ulikowski Dariusz; Chybowski Łucjan; Traczuk Piotr

    2015-01-01

    The aim of this study was to determine the impact of filamentous algae on the growth and survival of juvenile narrow-clawed crayfish, Astacus leptodactylus (Esch.), in rearing basins. Three stocking variants were used: A - basins with a layer of filamentous algae without imitation mineral substrate; B - basins with a layer of filamentous algae with imitation mineral substrate; C - basins without filamentous algae but with mineral substrate. The crayfish were reared from June 12 to October 10 ...

  20. Standing waves in a counter-rotating vortex filament pair

    Science.gov (United States)

    García-Azpeitia, Carlos

    2018-03-01

    The distance among two counter-rotating vortex filaments satisfies a beam-type of equation according to the model derived in [15]. This equation has an explicit solution where two straight filaments travel with constant speed at a constant distance. The boundary condition of the filaments is 2π-periodic. Using the distance of the filaments as bifurcating parameter, an infinite number of branches of periodic standing waves bifurcate from this initial configuration with constant rational frequency along each branch.

  1. Graphene-based filament material for thermal ionization

    Energy Technology Data Exchange (ETDEWEB)

    Hewitt, J. [Savannah River Site (SRS), Aiken, SC (United States). Savannah River National Lab. (SRNL); Shick, C. [Savannah River Site (SRS), Aiken, SC (United States). Savannah River National Lab. (SRNL); Siegfried, M. [Savannah River Site (SRS), Aiken, SC (United States). Savannah River National Lab. (SRNL)

    2017-09-19

    The use of graphene oxide materials for thermal ionization mass spectrometry analysis of plutonium and uranium has been investigated. Filament made from graphene oxide slurries have been 3-D printed. A method for attaching these filaments to commercial thermal ionization post assemblies has been devised. Resistive heating of the graphene based filaments under high vacuum showed stable operation in excess of 4 hours. Plutonium ion production has been observed in an initial set of filaments spiked with the Pu 128 Certified Reference Material.

  2. Nano-assembly of nanodiamonds by conjugation to actin filaments.

    Science.gov (United States)

    Bradac, Carlo; Say, Jana M; Rastogi, Ishan D; Cordina, Nicole M; Volz, Thomas; Brown, Louise J

    2016-03-01

    Fluorescent nanodiamonds (NDs) are remarkable objects. They possess unique mechanical and optical properties combined with high surface areas and controllable surface reactivity. They are non-toxic and hence suited for use in biological environments. NDs are also readily available and commercially inexpensive. Here, the exceptional capability of controlling and tailoring their surface chemistry is demonstrated. Small, bright diamond nanocrystals (size ˜30 nm) are conjugated to protein filaments of actin (length ˜3-7 µm). The conjugation to actin filaments is extremely selective and highly target-specific. These unique features, together with the relative simplicity of the conjugation-targeting method, make functionalised nanodiamonds a powerful and versatile platform in biomedicine and quantum nanotechnologies. Applications ranging from using NDs as superior biological markers to, potentially, developing novel bottom-up approaches for the fabrication of hybrid quantum devices that would bridge across the bio/solid-state interface are presented and discussed. © 2015 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  3. Can Morphing Methods Predict Intermediate Structures?

    Science.gov (United States)

    Weiss, Dahlia R.; Levitt, Michael

    2009-01-01

    Movement is crucial to the biological function of many proteins, yet crystallographic structures of proteins can give us only a static snapshot. The protein dynamics that are important to biological function often happen on a timescale that is unattainable through detailed simulation methods such as molecular dynamics as they often involve crossing high-energy barriers. To address this coarse-grained motion, several methods have been implemented as web servers in which a set of coordinates is usually linearly interpolated from an initial crystallographic structure to a final crystallographic structure. We present a new morphing method that does not extrapolate linearly and can therefore go around high-energy barriers and which can produce different trajectories between the same two starting points. In this work, we evaluate our method and other established coarse-grained methods according to an objective measure: how close a coarse-grained dynamics method comes to a crystallographically determined intermediate structure when calculating a trajectory between the initial and final crystal protein structure. We test this with a set of five proteins with at least three crystallographically determined on-pathway high-resolution intermediate structures from the Protein Data Bank. For simple hinging motions involving a small conformational change, segmentation of the protein into two rigid sections outperforms other more computationally involved methods. However, large-scale conformational change is best addressed using a nonlinear approach and we suggest that there is merit in further developing such methods. PMID:18996395

  4. NONLINEAR FORCE-FREE FIELD EXTRAPOLATION OF A CORONAL MAGNETIC FLUX ROPE SUPPORTING A LARGE-SCALE SOLAR FILAMENT FROM A PHOTOSPHERIC VECTOR MAGNETOGRAM

    Energy Technology Data Exchange (ETDEWEB)

    Jiang, Chaowei; Wu, S. T.; Hu, Qiang [Center for Space Plasma and Aeronomic Research, The University of Alabama in Huntsville, Huntsville, AL 35899 (United States); Feng, Xueshang, E-mail: cwjiang@spaceweather.ac.cn, E-mail: wus@uah.edu, E-mail: qh0001@uah.edu, E-mail: fengx@spaceweather.ac.cn [SIGMA Weather Group, State Key Laboratory for Space Weather, Center for Space Science and Applied Research, Chinese Academy of Sciences, Beijing 100190 (China)

    2014-05-10

    Solar filaments are commonly thought to be supported in magnetic dips, in particular, in those of magnetic flux ropes (FRs). In this Letter, based on the observed photospheric vector magnetogram, we implement a nonlinear force-free field (NLFFF) extrapolation of a coronal magnetic FR that supports a large-scale intermediate filament between an active region and a weak polarity region. This result is a first, in the sense that current NLFFF extrapolations including the presence of FRs are limited to relatively small-scale filaments that are close to sunspots and along main polarity inversion lines (PILs) with strong transverse field and magnetic shear, and the existence of an FR is usually predictable. In contrast, the present filament lies along the weak-field region (photospheric field strength ≲ 100 G), where the PIL is very fragmented due to small parasitic polarities on both sides of the PIL and the transverse field has a low signal-to-noise ratio. Thus, extrapolating a large-scale FR in such a case represents a far more difficult challenge. We demonstrate that our CESE-MHD-NLFFF code is sufficient for the challenge. The numerically reproduced magnetic dips of the extrapolated FR match observations of the filament and its barbs very well, which strongly supports the FR-dip model for filaments. The filament is stably sustained because the FR is weakly twisted and strongly confined by the overlying closed arcades.

  5. Fabrication of Polylactide Nanocomposite Filament Using Melt Extrusion and Filament Characterization for 3D Printing

    Science.gov (United States)

    Jain, Shrenik Kumar

    Fused deposition modeling (FDM) technology uses thermoplastic filament for layer by layer fabrication of objects. To make functional objects with desired properties, composite filaments are required in the FDM. In this thesis, less expensive mesoporous Nano carbon (NC) and carbon nanotube (CNT) infused in Polylactide (PLA) thermoplastic filaments were fabricated to improve the electrical properties and maintain sufficient strength for 3D printing. Solution blending was used for nanocomposite fabrication and melt extrusion was employed to make cylindrical filaments. Mechanical and electrical properties of 1 to 20 wt% of NC and 1 to 3 wt% of CNT filaments were investigated and significant improvement of conductivity (3.76 S/m) and sufficient yield strength (35MPa) were obtained. Scanning electron microscopy (SEM) images exhibited uniform dispersion of nanoparticles in polymer matrix and differential scanning calorimetry (DSC) results showed no significant changes in the glass transition temperature (Tg) for all the compositions. Perspective uses of this filament are for fabrication of electrical wires in 3D printed robots, drones, prosthetics, orthotics and others.

  6. A penny-shaped crack in a filament-reinforced matrix. I - The filament model. II - The crack problem

    Science.gov (United States)

    Erdogan, F.; Pacella, A. H.

    1974-01-01

    The study deals with the elastostatic problem of a penny-shaped crack in an elastic matrix which is reinforced by filaments or fibers perpendicular to the plane of the crack. An elastic filament model is first developed, followed by consideration of the application of the model to the penny-shaped crack problem in which the filaments of finite length are asymmetrically distributed around the crack. Since the primary interest is in the application of the results to studies relating to the fracture of fiber or filament-reinforced composites and reinforced concrete, the main emphasis of the study is on the evaluation of the stress intensity factor along the periphery of the crack, the stresses in the filaments or fibers, and the interface shear between the matrix and the filaments or fibers. Using the filament model developed, the elastostatic interaction problem between a penny-shaped crack and a slender inclusion or filament in an elastic matrix is formulated.

  7. Partially folded intermediates during trypsinogen denaturation

    Directory of Open Access Journals (Sweden)

    Martins N.F.

    1999-01-01

    Full Text Available The equilibrium unfolding of bovine trypsinogen was studied by circular dichroism, differential spectra and size exclusion HPLC. The change in free energy of denaturation was = 6.99 ± 1.40 kcal/mol for guanidine hydrochloride and = 6.37 ± 0.57 kcal/mol for urea. Satisfactory fits of equilibrium unfolding transitions required a three-state model involving an intermediate in addition to the native and unfolded forms. Size exclusion HPLC allowed the detection of an intermediate population of trypsinogen whose Stokes radii varied from 24.1 ± 0.4 Å to 26.0 ± 0.3 Å for 1.5 M and 2.5 M guanidine hydrochloride, respectively. During urea denaturation, the range of Stokes radii varied from 23.9 ± 0.3 Å to 25.7 ± 0.6 Å for 4.0 M and 6.0 M urea, respectively. Maximal intrinsic fluorescence was observed at about 3.8 M urea with 8-aniline-1-naphthalene sulfonate (ANS binding. These experimental data indicate that the unfolding of bovine trypsinogen is not a simple transition and suggest that the equilibrium intermediate population comprises one intermediate that may be characterized as a molten globule. To obtain further insight by studying intermediates representing different stages of unfolding, we hope to gain a better understanding of the complex interrelations between protein conformation and energetics.

  8. Synthesis and functionalization of coiled carbon filaments

    Science.gov (United States)

    Hikita, Muneaki

    Coiled carbon filaments have one of the most attractive three-dimensional forms in carbon materials due to their helical morphologies. Because of their shape and carbon structure, they exhibit excellent mechanical and electrical properties such as superelasticity, low Young's modulus, relatively high electrical conductivity, and good electromagnetic (EM) wave absorption. Therefore, they are good candidates as fillers in composite materials for tactile sensor and electromagnetic interference shielding. In medical areas of interests, coiled carbon filaments can be used as micro and nano heaters or trigger for thermotherapy and biosensors using EM wave exposure because absorbed EM waves by coiled carbon filaments are converted into heat. Although various shapes of coiled carbon filaments have been discovered, optimum synthesis conditions and growth mechanisms of coiled carbon filaments are poorly understood. The study of growth kinetics is significant not only to analyze catalyst activity but also to establish the growth mechanisms of coiled carbon filaments. The establishment of growth mechanisms would be useful for determining optimum synthesis conditions and maximizing the quantity of carbon filaments synthesized for a given application. In the first study, tip grown single helical carbon filaments or carbon nanocoils (CNCs) were synthesized by a chemical vapor deposition method using tin-iron-oxide (Sn-Fe-O) xerogel film catalyst. The Sn-Fe-O catalyst was prepared by a low-cost sol-gel method using stannous acetate and ferric acetate as precursors. The growth kinetics of CNCs were monitored by a thermogravimetric analyzer, and the experimental result was correlated using a one-dimensional kinetic model, corresponding to one-dimensional tip growth. In the second study, bidirectionally grown double helical filaments or carbon microcoils (CMCs) were synthesized using a chemical vapor deposition method. CMCs obtained at two reaction temperatures were compared. CMCs

  9. Leiomodin-3 dysfunction results in thin filament disorganization and nemaline myopathy.

    Science.gov (United States)

    Yuen, Michaela; Sandaradura, Sarah A; Dowling, James J; Kostyukova, Alla S; Moroz, Natalia; Quinlan, Kate G; Lehtokari, Vilma-Lotta; Ravenscroft, Gianina; Todd, Emily J; Ceyhan-Birsoy, Ozge; Gokhin, David S; Maluenda, Jérome; Lek, Monkol; Nolent, Flora; Pappas, Christopher T; Novak, Stefanie M; D'Amico, Adele; Malfatti, Edoardo; Thomas, Brett P; Gabriel, Stacey B; Gupta, Namrata; Daly, Mark J; Ilkovski, Biljana; Houweling, Peter J; Davidson, Ann E; Swanson, Lindsay C; Brownstein, Catherine A; Gupta, Vandana A; Medne, Livija; Shannon, Patrick; Martin, Nicole; Bick, David P; Flisberg, Anders; Holmberg, Eva; Van den Bergh, Peter; Lapunzina, Pablo; Waddell, Leigh B; Sloboda, Darcée D; Bertini, Enrico; Chitayat, David; Telfer, William R; Laquerrière, Annie; Gregorio, Carol C; Ottenheijm, Coen A C; Bönnemann, Carsten G; Pelin, Katarina; Beggs, Alan H; Hayashi, Yukiko K; Romero, Norma B; Laing, Nigel G; Nishino, Ichizo; Wallgren-Pettersson, Carina; Melki, Judith; Fowler, Velia M; MacArthur, Daniel G; North, Kathryn N; Clarke, Nigel F

    2014-11-01

    Nemaline myopathy (NM) is a genetic muscle disorder characterized by muscle dysfunction and electron-dense protein accumulations (nemaline bodies) in myofibers. Pathogenic mutations have been described in 9 genes to date, but the genetic basis remains unknown in many cases. Here, using an approach that combined whole-exome sequencing (WES) and Sanger sequencing, we identified homozygous or compound heterozygous variants in LMOD3 in 21 patients from 14 families with severe, usually lethal, NM. LMOD3 encodes leiomodin-3 (LMOD3), a 65-kDa protein expressed in skeletal and cardiac muscle. LMOD3 was expressed from early stages of muscle differentiation; localized to actin thin filaments, with enrichment near the pointed ends; and had strong actin filament-nucleating activity. Loss of LMOD3 in patient muscle resulted in shortening and disorganization of thin filaments. Knockdown of lmod3 in zebrafish replicated NM-associated functional and pathological phenotypes. Together, these findings indicate that mutations in the gene encoding LMOD3 underlie congenital myopathy and demonstrate that LMOD3 is essential for the organization of sarcomeric thin filaments in skeletal muscle.

  10. Fabrication of PLA Filaments and its Printable Performance

    Science.gov (United States)

    Liu, Wenjie; Zhou, Jianping; Ma, Yuming; Wang, Jie; Xu, Jie

    2017-12-01

    Fused deposition modeling (FDM) is a typical 3D printing technology and preparation of qualified filaments is the basis. In order to prepare polylactic acid (PLA) filaments suitable for personalized FDM 3D printing, this article investigated the effect of factors such as extrusion temperature and screw speed on the diameter, surface roughness and ultimate tensile stress of the obtained PLA filaments. The optimal process parameters for fabrication of qualified filaments were determined. Further, the printable performance of the obtained PLA filaments for 3D objects was preliminarily explored.

  11. Apoptotic-like programed cell death in fungi: the benefits in filamentous species

    International Nuclear Information System (INIS)

    Shlezinger, Neta; Goldfinger, Nir; Sharon, Amir

    2012-01-01

    Studies conducted in the early 1990s showed for the first time that Saccharomyces cerevisiae can undergo cell death with hallmarks of animal apoptosis. These findings came as a surprise, since suicide machinery was unexpected in unicellular organisms. Today, apoptosis in yeast is well-documented. Apoptotic death of yeast cells has been described under various conditions and S. cerevisiae homologs of human apoptotic genes have been identified and characterized. These studies also revealed fundamental differences between yeast and animal apoptosis; in S. cerevisiae apoptosis is mainly associated with aging and stress adaptation, unlike animal apoptosis, which is essential for proper development. Further, many apoptosis regulatory genes are either missing, or highly divergent in S. cerevisiae. Therefore, in this review we will use the term apoptosis-like programed cell death (PCD) instead of apoptosis. Despite these significant differences, S. cerevisiae has been instrumental in promoting the study of heterologous apoptotic proteins, particularly from human. Work in fungi other than S. cerevisiae revealed differences in the manifestation of PCD in single cell (yeasts) and multicellular (filamentous) species. Such differences may reflect the higher complexity level of filamentous species, and hence the involvement of PCD in a wider range of processes and life styles. It is also expected that differences might be found in the apoptosis apparatus of yeast and filamentous species. In this review we focus on aspects of PCD that are unique or can be better studied in filamentous species. We will highlight the similarities and differences of the PCD machinery between yeast and filamentous species and show the value of using S. cerevisiae along with filamentous species to study apoptosis.

  12. Structural modeling and molecular dynamics simulation of the actin filament.

    Science.gov (United States)

    Splettstoesser, Thomas; Holmes, Kenneth C; Noé, Frank; Smith, Jeremy C

    2011-07-01

    Actin is a major structural protein of the eukaryotic cytoskeleton and enables cell motility. Here, we present a model of the actin filament (F-actin) that not only incorporates the global structure of the recently published model by Oda et al. but also conserves internal stereochemistry. A comparison is made using molecular dynamics simulation of the model with other recent F-actin models. A number of structural determents such as the protomer propeller angle, the number of hydrogen bonds, and the structural variation among the protomers are analyzed. The MD comparison is found to reflect the evolution in quality of actin models over the last 6 years. In addition, simulations of the model are carried out in states with both ADP or ATP bound and local hydrogen-bonding differences characterized. Copyright © 2011 Wiley-Liss, Inc.

  13. How filamentous plant pathogen effectors are translocated to host cells.

    Science.gov (United States)

    Lo Presti, Libera; Kahmann, Regine

    2017-08-01

    The interaction of microbes with "signature" plants is largely governed by secreted effector proteins, which serve to dampen plant defense responses and modulate host cell processes. Secreted effectors can function either in the apoplast or within plant cell compartments. How oomycetes and fungi translocate their effectors to plant cells is still poorly understood and controversial. While most oomycete effectors share a common 'signature' that was proposed to mediate their uptake via endocytosis, fungal effectors display no conserved motifs at the primary amino acid sequence level. Here we summarize current knowledge in the field of oomycete and fungal effector uptake and highlight emerging themes that may unite rather than set apart these unrelated filamentous pathogens. Copyright © 2017 Elsevier Ltd. All rights reserved.

  14. Molecular Mechanisms Regulating Sporulation in the Filamentous Fungus Ashbya gossypii

    DEFF Research Database (Denmark)

    Wasserstrom, Lisa

    , which is regulated by the pheromone response pathway. Most ascomycetes have been reported to produce meiotic spores, however, a sexual cycle has not yet been identified in the filamentous fungus Ashbya gossypii. The main focus of my doctoral thesis has therefore been to understand the mechanisms behind...... sporulation in this fungus.  The lifecycle of A. gossypii starting with a haploid spore that matures into spore-containing mycelia can be completed without the need for a mating partner. Spores in A. gossypii are thought to be derived sexually like all other Saccharomycetaceae species, but the sexual cycle...... suggesting that other proteins generate DSBs in this fungus. In summary, this work has led to better understanding of the components regulating sporulation in A. gossypii and their hierarchical organization....

  15. Morphogenesis of filaments growing in flexible confinements

    Science.gov (United States)

    Vetter, R.; Wittel, F. K.; Herrmann, H. J.

    2014-07-01

    Space-saving design is a requirement that is encountered in biological systems and the development of modern technological devices alike. Many living organisms dynamically pack their polymer chains, filaments or membranes inside deformable vesicles or soft tissue-like cell walls, chorions and buds. Surprisingly little is known about morphogenesis due to growth in flexible confinements—perhaps owing to the daunting complexity lying in the nonlinear feedback between packed material and expandable cavity. Here we show by experiments and simulations how geometric and material properties lead to a plethora of morphologies when elastic filaments are growing far beyond the equilibrium size of a flexible thin sheet they are confined in. Depending on friction, sheet flexibility and thickness, we identify four distinct morphological phases emerging from bifurcation and present the corresponding phase diagram. Four order parameters quantifying the transitions between these phases are proposed.

  16. SWAYING THREADS OF A SOLAR FILAMENT

    International Nuclear Information System (INIS)

    Lin, Y.; Engvold, O.; Langangen, Oe.; Rouppe van der Voort, L. H. M.; Soler, R.; Ballester, J. L.; Oliver, R.

    2009-01-01

    From recent high-resolution observations obtained with the Swedish 1 m Solar Telescope in La Palma, we detect swaying motions of individual filament threads in the plane of the sky. The oscillatory characters of these motions are comparable with oscillatory Doppler signals obtained from corresponding filament threads. Simultaneous recordings of motions in the line of sight and in the plane of the sky give information about the orientation of the oscillatory plane. These oscillations are interpreted in the context of the magnetohydrodynamic (MHD) theory. Kink MHD waves supported by the thread body are proposed as an explanation of the observed thread oscillations. On the basis of this interpretation and by means of seismological arguments, we give an estimation of the thread Alfven speed and magnetic field strength by means of seismological arguments.

  17. Helicity and Filament Channels? The Straight Twist!

    Science.gov (United States)

    Antiochos, Spiro K.

    2010-01-01

    One of the most important and most puzzling features of the coronal magnetic field is that it appears to have smooth magnetic structure with little evidence for non-potentiality except at special locations, photospheric polarity inversions lines where the non-potentiality is observed as a filament channel. This characteristic feature of the closed-field corona is highly unexpected given that photospheric motions continuously tangle its magnetic field. Although reconnection can eliminate some of the injected structure, it cannot destroy the helicity, which should build up to produce observable complexity. We propose that an inverse cascade process transports the injected helicity from the interior of closed flux regions to their boundaries, polarity inversion lines, creating filament channels. We describe how the helicity is injected and transported and calculate the relevant rates. We argue that one process, helicity transport, can explain both the observed lack and presence of structure in the coronal magnetic field.

  18. Morgellons disease: a filamentous borrelial dermatitis.

    Science.gov (United States)

    Middelveen, Marianne J; Stricker, Raphael B

    2016-01-01

    Morgellons disease (MD) is a dermopathy characterized by multicolored filaments that lie under, are embedded in, or project from skin. Although MD was initially considered to be a delusional disorder, recent studies have demonstrated that the dermopathy is associated with tickborne infection, that the filaments are composed of keratin and collagen, and that they result from proliferation of keratinocytes and fibroblasts in epithelial tissue. Culture, histopathological and molecular evidence of spirochetal infection associated with MD has been presented in several published studies using a variety of techniques. Spirochetes genetically identified as Borrelia burgdorferi sensu stricto predominate as the infective agent in most of the Morgellons skin specimens studied so far. Other species of Borrelia including Borrelia garinii , Borrelia miyamotoi , and Borrelia hermsii have also been detected in skin specimens taken from MD patients. The optimal treatment for MD remains to be determined.

  19. Statistical study of solar filaments since 1919

    Science.gov (United States)

    Aboudarham, Jean

    2016-04-01

    Science board of Paris Observatory funded the data capture of tables associated with Meudon synoptic maps of Solar activity, which were published for observations ranging from 1919 to 1992. The EU HELIO project developed automatic recognition codes, especially concerning filaments based on observations between 1996 en 2014 (and soon, up to now). We plan to fill the gap between the two catalogues in the short term. But it is already possible to study filaments behavior over quite long periods of time. We present here the first series of results obtained from this analysis which give some clue about the way Solar activity behaves in various parts of the cycle, and about the way if depends on the hemisphere where activity occurs. This information could then be correlated with events catalogues (e.g. flares, CMEs, …) in order to link those phenomena with concrete Solar activity.

  20. Actin organization and dynamics in filamentous fungi.

    Science.gov (United States)

    Berepiki, Adokiye; Lichius, Alexander; Read, Nick D

    2011-11-02

    Growth and morphogenesis of filamentous fungi is underpinned by dynamic reorganization and polarization of the actin cytoskeleton. Actin has crucial roles in exocytosis, endocytosis, organelle movement and cytokinesis in fungi, and these processes are coupled to the production of distinct higher-order structures (actin patches, cables and rings) that generate forces or serve as tracks for intracellular transport. New approaches for imaging actin in living cells are revealing important similarities and differences in actin architecture and organization within the fungal kingdom, and have yielded key insights into cell polarity, tip growth and long-distance intracellular transport. In this Review, we discuss the contribution that recent live-cell imaging and mutational studies have made to our understanding of the dynamics and regulation of actin in filamentous fungi.

  1. Laser filamentation mathematical methods and models

    CERN Document Server

    Lorin, Emmanuel; Moloney, Jerome

    2016-01-01

    This book is focused on the nonlinear theoretical and mathematical problems associated with ultrafast intense laser pulse propagation in gases and in particular, in air. With the aim of understanding the physics of filamentation in gases, solids, the atmosphere, and even biological tissue, specialists in nonlinear optics and filamentation from both physics and mathematics attempt to rigorously derive and analyze relevant non-perturbative models. Modern laser technology allows the generation of ultrafast (few cycle) laser pulses, with intensities exceeding the internal electric field in atoms and molecules (E=5x109 V/cm or intensity I = 3.5 x 1016 Watts/cm2 ). The interaction of such pulses with atoms and molecules leads to new, highly nonlinear nonperturbative regimes, where new physical phenomena, such as High Harmonic Generation (HHG), occur, and from which the shortest (attosecond - the natural time scale of the electron) pulses have been created. One of the major experimental discoveries in this nonlinear...

  2. Motility patterns of filamentous sulfur bacteria, Beggiatoa spp

    DEFF Research Database (Denmark)

    Dunker, Rita; Røy, Hans; Kamp, Anja

    2011-01-01

    The large sulfur bacteria, Beggiatoa spp., live on the oxidation of sulfide with oxygen or nitrate, but avoid high concentrations of both sulfide and oxygen. As gliding filaments, they rely on reversals in the gliding direction to find their preferred environment, the oxygen–sulfide interface. We...... observed the chemotactic patterns of single filaments in a transparent agar medium and scored their reversals and the glided distances between reversals. Filaments within the preferred microenvironment glided distances shorter than their own length between reversals that anchored them in their position...... as a microbial mat. Filaments in the oxic region above the mat or in the sulfidic, anoxic region below the mat glided distances longer than the filament length between reversals. This reversal behavior resulted in a diffusion-like spreading of the filaments. A numerical model of such gliding filaments...

  3. Viruses: incredible nanomachines. New advances with filamentous phages.

    Science.gov (United States)

    Hemminga, Marcus A; Vos, Werner L; Nazarov, Petr V; Koehorst, Rob B M; Wolfs, Cor J A M; Spruijt, Ruud B; Stopar, David

    2010-03-01

    During recent decades, bacteriophages have been at the cutting edge of new developments in molecular biology, biophysics, and, more recently, bionanotechnology. In particular filamentous viruses, for example bacteriophage M13, have a virion architecture that enables precision building of ordered and defect-free two and three-dimensional structures on a nanometre scale. This could not have been possible without detailed knowledge of coat protein structure and dynamics during the virus reproduction cycle. The results of the spectroscopic studies conducted in our group compellingly demonstrate a critical role of membrane embedment of the protein both during infectious entry of the virus into the host cell and during assembly of the new virion in the host membrane. The protein is effectively embedded in the membrane by a strong C-terminal interfacial anchor, which together with a simple tilt mechanism and a subtle structural adjustment of the extreme end of its N terminus provides favourable thermodynamical association of the protein in the lipid bilayer. This basic physicochemical rule cannot be violated and any new bionanotechnology that will emerge from bacteriophage M13 should take this into account.

  4. Laser induced white lighting of tungsten filament

    Science.gov (United States)

    Strek, W.; Tomala, R.; Lukaszewicz, M.

    2018-04-01

    The sustained bright white light emission of thin tungsten filament was induced under irradiation with focused beam of CW infrared laser diode. The broadband emission centered at 600 nm has demonstrated the threshold behavior on excitation power. Its intensity increased non-linearly with excitation power. The emission occurred only from the spot of focused beam of excitation laser diode. The white lighting was accompanied by efficient photocurrent flow and photoelectron emission which both increased non-linearly with laser irradiation power.

  5. Filamented plasmas in laser ablation of solids

    Czech Academy of Sciences Publication Activity Database

    Davies, J.R.; Fajardo, M.; Kozlová, Michaela; Mocek, Tomáš; Polan, Jiří; Rus, Bedřich

    2009-01-01

    Roč. 51, č. 3 (2009), 035013/1-035013/12 ISSN 0741-3335 EU Projects: European Commission(XE) 12843 - TUIXS Grant - others:FCT(PT) POCI/FIS/59563/2004 Institutional research plan: CEZ:AV0Z10100523 Keywords : magneto-hydrodynamic modelling * perturbation * filaments * x-ray * plasma Subject RIV: BH - Optics, Masers, Lasers Impact factor: 2.409, year: 2009

  6. A first approach to filament dynamics

    Energy Technology Data Exchange (ETDEWEB)

    Silva, P E S; De Abreu, F Vistulo; Dias, R G [Department of Physics, University of Aveiro (Portugal); Simoes, R, E-mail: fva@ua.p [I3N-Institute for Nanostructures, Nanomodelling and Nanofabrication (Portugal)

    2010-11-15

    Modelling elastic filament dynamics is a topic of high interest due to the wide range of applications. However, it has reached a high level of complexity in the literature, making it unaccessible to a beginner. In this paper we explain the main steps involved in the computational modelling of the dynamics of an elastic filament. We first derive equations governing the dynamics of an elastic lament suitable for a computer simulation implementation. The derivation starts from the relation between forces and potential energy in conservative systems in order to derive the equation of motion of any bead in the filament. Only two-dimensional movements are considered, but extensions to three dimensions can follow similar lines. Suggestions for computer implementations are provided in Matlab as well as an example of application related to the generation of musical sounds. This example allows a critical analysis of the numerical results obtained using a cross-disciplinary perspective. Since derivations start from basic physics equations, use simple calculus and computational implementations are straightforward, this paper proposes a different approach to introduce simple molecular dynamics simulations or animations of real systems in undergraduate elasticity or computer modelling courses.

  7. The Magnetic Structure of Filament Barbs

    Science.gov (United States)

    Chae, Jongchul; Moon, Yong-Jae; Park, Young-Deuk

    2005-06-01

    There is a controversy about how features protruding laterally from filaments, called barbs, are magnetically structured. On 2004 August 3, we observed a filament that had well-developed barbs. The observations were performed using the 10 inch refractor of the Big Bear Solar Observatory. A fast camera was employed to capture images at five different wavelengths of the Hα line and successively record them on the basis of frame selection. The terminating points of the barbs were clearly discernable in the Hα images without any ambiguity. The comparison of the Hα images with the magnetograms taken by SOHO MDI revealed that the termination occurred above the minor polarity inversion line dividing the magnetic elements of the major polarity and those of the minor polarity. There is also evidence that the flux cancellation proceeded on the polarity inversion line. Our results together with similar other recent observations support the idea that filament barbs are cool matter suspended in local dips of magnetic field lines, formed by magnetic reconnection in the chromosphere.

  8. Tracer filamentation at an unstable ocean front

    Science.gov (United States)

    Feng, Yen Chia; Mahadevan, Amala; Thiffeault, Jean-Luc; Yecko, Philip

    2017-11-01

    A front, where two bodies of ocean water with different physical properties meet, can become unstable and lead to a flow with high strain rate and vorticity. Phytoplankton and other oceanic tracers are stirred into filaments by such flow fields, as can often be seen in satellite imagery. The stretching and folding of a tracer by a two-dimensional flow field has been well studied. In the ocean, however, the vertical shear of horizontal velocity is typically two orders of magnitude larger than the horizontal velocity gradient. Theoretical calculations show that vertical shear alters the way in which horizontal strain affects the tracer, resulting in thin, sloping structures in the tracer field. Using a non-hydrostatic ocean model of an unstable ocean front, we simulate tracer filamentation to identify the effect of vertical shear on the deformation of the tracer. In a complementary laboratory experiment, we generate a simple, vertically sheared strain flow and use dye and particle image velocimetry to quantify the filamentary structures in terms of the strain and shear. We identify how vertical shear alters the tracer filaments and infer how the evolution of tracers in the ocean will differ from the idealized two-dimensional paradigm. Support of NSF DMS-1418956 is acknowledged.

  9. A first approach to filament dynamics

    International Nuclear Information System (INIS)

    Silva, P E S; De Abreu, F Vistulo; Dias, R G; Simoes, R

    2010-01-01

    Modelling elastic filament dynamics is a topic of high interest due to the wide range of applications. However, it has reached a high level of complexity in the literature, making it unaccessible to a beginner. In this paper we explain the main steps involved in the computational modelling of the dynamics of an elastic filament. We first derive equations governing the dynamics of an elastic lament suitable for a computer simulation implementation. The derivation starts from the relation between forces and potential energy in conservative systems in order to derive the equation of motion of any bead in the filament. Only two-dimensional movements are considered, but extensions to three dimensions can follow similar lines. Suggestions for computer implementations are provided in Matlab as well as an example of application related to the generation of musical sounds. This example allows a critical analysis of the numerical results obtained using a cross-disciplinary perspective. Since derivations start from basic physics equations, use simple calculus and computational implementations are straightforward, this paper proposes a different approach to introduce simple molecular dynamics simulations or animations of real systems in undergraduate elasticity or computer modelling courses.

  10. Magnetization Modeling of Twisted Superconducting Filaments

    CERN Document Server

    Satiramatekul, T; Devred, Arnaud; Leroy, Daniel

    2007-01-01

    This paper presents a new Finite Element numerical method to analyze the coupling between twisted filaments in a superconducting multifilament composite wire. To avoid the large number of elements required by a 3D code, the proposed method makes use of the energy balance principle in a 2D code. The relationship between superconductor critical current density and local magnetic flux density is implemented in the program for the Bean and modified Kim models. The modeled wire is made up of six filaments twisted together and embedded in a lowresistivity matrix. Computations of magnetization cycle and of the electric field pattern have been performed for various twist pitch values in the case of a pure copper matrix. The results confirm that the maximum magnetization depends on the matrix conductivity, the superconductor critical current density, the applied field frequency, and the filament twist pitch. The simulations also lead to a practical criterion for wire design that can be used to assess whether or not th...

  11. On the fragmentation of filaments in a molecular cloud simulation

    Science.gov (United States)

    Chira, R.-A.; Kainulainen, J.; Ibáñez-Mejía, J. C.; Henning, Th.; Mac Low, M.-M.

    2018-03-01

    Context. The fragmentation of filaments in molecular clouds has attracted a lot of attention recently as there seems to be a close relation between the evolution of filaments and star formation. The study of the fragmentation process has been motivated by simple analytical models. However, only a few comprehensive studies have analysed the evolution of filaments using numerical simulations where the filaments form self-consistently as part of large-scale molecular cloud evolution. Aim. We address the early evolution of parsec-scale filaments that form within individual clouds. In particular, we focus on three questions: How do the line masses of filaments evolve? How and when do the filaments fragment? How does the fragmentation relate to the line masses of the filaments? Methods: We examine three simulated molecular clouds formed in kiloparsec-scale numerical simulations performed with the FLASH adaptive mesh refinement magnetohydrodynamic code. The simulations model a self-gravitating, magnetised, stratified, supernova-driven interstellar medium, including photoelectric heating and radiative cooling. We follow the evolution of the clouds for 6 Myr from the time self-gravity starts to act. We identify filaments using the DisPerSe algorithm, and compare the results to other filament-finding algorithms. We determine the properties of the identified filaments and compare them with the predictions of analytic filament stability models. Results: The average line masses of the identified filaments, as well as the fraction of mass in filamentary structures, increases fairly continuously after the onset of self-gravity. The filaments show fragmentation starting relatively early: the first fragments appear when the line masses lie well below the critical line mass of Ostriker's isolated hydrostatic equilibrium solution ( 16 M⊙ pc-1), commonly used as a fragmentation criterion. The average line masses of filaments identified in three-dimensional volume density cubes

  12. Cholera toxin can catalyze ADP-ribosylation of cytoskeletal proteins

    Energy Technology Data Exchange (ETDEWEB)

    Kaslow, H.R.; Groppi, V.E.; Abood, M.E.; Bourne, H.R.

    1981-11-01

    Cholera toxin catalyzes transfer of radiolabel from (/sup 32/P)NAD/sup +/ to several peptides in particulate preparations of human foreskin fibroblasts. Resolution of these peptides by two-dimensional gel electrophoresis allowed identification of two peptides of M/sub r/ = 42,000 and 52,000 as peptide subunits of a regulatory component of adenylate cyclase. The radiolabeling of another group of peptides (M/sub r/ = 50,000 to 65,000) suggested that cholera toxin could catalyze ADP-ribosylation of cytoskeletal proteins. This suggestion was confirmed by showing that incubation with cholera toxin and (/sup 32/P)NAD/sup +/ caused radiolabeling of purified microtubule and intermediate filament proteins.

  13. Isolation and characterization of flagellar filament from zoospores of Dermatophilus congolensis.

    Science.gov (United States)

    Hiraizumi, Mieko; Tagawa, Yuichi

    2014-09-17

    Highly motile zoospores from Dermatophilus congolensis bovine isolates from clinical dermatophilosis in Japan were obtained by culturing at 27°C in an ambient atmosphere on heart infusion agar supplemented with 5% defibrinated sheep blood for 72h or in heart infusion broth for 48h with gentle shaking. After vigorous mechanical agitation of the zoospore suspension, the flagellar filaments detached from motile zoospores and were isolated in the clear gelatinous part of the final pellet by differential centrifugation. Typical morphology of a flagellar filament, with a width of approximately 15nm, was observed in the isolated flagellar filament by electron microscopy. A single major protein (flagellin) band with an apparent molecular mass of 35kDa was detected in the flagellar filament of D. congolensis strain AM-1 and that of 33kDa was detected in strain IT-2 by SDS-PAGE. In immunoblot analysis of whole-cell proteins from seven isolates of D. congolensis, antiserum to strain AM-1 zoospores reacted with the 35-kDa antigen band of strain AM-1, but not with any antigen band of other strains in a similar molecular mass range. In contrast, antiserum to strain IT-2 zoospores reacted with antigen bands at 33kDa from six strains, except strain AM-1. Similar strain-specific reactions of these anti-zoospore sera with isolated flagellar filaments from strains AM-1 and IT-2 were confirmed by immunoblot, indicating the presence of antigenic variations of flagellins of D. congolensis zoospores. Copyright © 2014 Elsevier B.V. All rights reserved.

  14. an intermediate moisture meat

    African Journals Online (AJOL)

    STORAGESEVER

    2008-07-04

    Jul 4, 2008 ... thereby increasing their animal protein intake, this experiment therefore become imperative. Meat from the semimembranosus (SM), biceps .... most critical eating quality that determines whether consumers are repeat buyer. It is however worthy of note that as a result of the heat treatment and the use of.

  15. Measuring Filament Orientation: A New Quantitative, Local Approach

    Energy Technology Data Exchange (ETDEWEB)

    Green, C.-E.; Cunningham, M. R.; Jones, P. A. [School of Physics, University of New South Wales, Sydney, NSW, 2052 (Australia); Dawson, J. R. [CSIRO Astronomy and Space Science, Australia Telescope National Facility, P.O. Box 76, Epping, NSW 1710 (Australia); Novak, G. [Center for Interdisciplinary Exploration and Research in Astrophysics (CIERA) and Department of Physics and Astronomy, Northwestern University, 2145 Sheridan Road, Evanston, IL 60208 (United States); Fissel, L. M. [National Radio Astronomy Observatory (NRAO), 520 Edgemont Road, Charlottesville, VA, 22903 (United States)

    2017-09-01

    The relative orientation between filamentary structures in molecular clouds and the ambient magnetic field provides insight into filament formation and stability. To calculate the relative orientation, a measurement of filament orientation is first required. We propose a new method to calculate the orientation of the one-pixel-wide filament skeleton that is output by filament identification algorithms such as filfinder. We derive the local filament orientation from the direction of the intensity gradient in the skeleton image using the Sobel filter and a few simple post-processing steps. We call this the “Sobel-gradient method.” The resulting filament orientation map can be compared quantitatively on a local scale with the magnetic field orientation map to then find the relative orientation of the filament with respect to the magnetic field at each point along the filament. It can also be used for constructing radial profiles for filament width fitting. The proposed method facilitates automation in analyses of filament skeletons, which is imperative in this era of “big data.”.

  16. Patterns of molecular motors that guide and sort filaments.

    Science.gov (United States)

    Rupp, Beat; Nédélec, François

    2012-11-21

    Molecular motors can be immobilized to transport filaments and loads that are attached to these filaments inside a nano-device. However, if motors are distributed uniformly over a flat surface, the motility is undirected, and the filaments move equally in all directions. For many applications it is important to control the direction in which the filaments move, and two strategies have been explored to achieve this: applying external forces and confining the filaments inside channels. In this article, we discuss a third strategy in which the topography of the sample remains flat, but the motors are distributed non-uniformly over the surface. Systems of filaments and patterned molecular motors were simulated using a stochastic engine that included Brownian motion and filament bending elasticity. Using an evolutionary algorithm, patterns were optimized for their capacity to precisely control the paths of the filaments. We identified patterns of motors that could either direct the filaments in a particular direction, or separate short and long filaments. These functionalities already exceed what has been achieved with confinement. The patterns are composed of one or two types of motors positioned in lines or along arcs and should be easy to manufacture. Finally, these patterns can be easily combined into larger designs, allowing one to precisely control the motion of microscopic objects inside a device.

  17. Information acquisition and financial intermediation

    OpenAIRE

    Boyarchenko, Nina

    2012-01-01

    This paper considers the problem of information acquisition in an intermediated market, where the specialists have access to superior technology for acquiring information. These informational advantages of specialists relative to households lead to disagreement between the two groups, changing the shape of the intermediation-constrained region of the economy and increasing the frequency of periods when the intermediation constraint binds. Acquiring the additional information is, however, cost...

  18. Analysis of differential protein expression in normal and neoplastic human breast epithelial cell lines

    Energy Technology Data Exchange (ETDEWEB)

    Williams, K.; Chubb, C.; Huberman, E.; Giometti, C.S.

    1997-07-01

    High resolution two dimensional get electrophoresis (2DE) and database analysis was used to establish protein expression patterns for cultured normal human mammary epithelial cells and thirteen breast cancer cell lines. The Human Breast Epithelial Cell database contains the 2DE protein patterns, including relative protein abundances, for each cell line, plus a composite pattern that contains all the common and specifically expressed proteins from all the cell lines. Significant differences in protein expression, both qualitative and quantitative, were observed not only between normal cells and tumor cells, but also among the tumor cell lines. Eight percent of the consistently detected proteins were found in significantly (P < 0.001) variable levels among the cell lines. Using a combination of immunostaining, comigration with purified protein, subcellular fractionation, and amino-terminal protein sequencing, we identified a subset of the differentially expressed proteins. These identified proteins include the cytoskeletal proteins actin, tubulin, vimentin, and cytokeratins. The cell lines can be classified into four distinct groups based on their intermediate filament protein profile. We also identified heat shock proteins; hsp27, hsp60, and hsp70 varied in abundance and in some cases in the relative phosphorylation levels among the cell lines. Finally, we identified IMP dehydrogenase in each of the cell lines, and found the levels of this enzyme in the tumor cell lines elevated 2- to 20-fold relative to the levels in normal cells.

  19. Intermediate inputs and economic productivity.

    Science.gov (United States)

    Baptist, Simon; Hepburn, Cameron

    2013-03-13

    Many models of economic growth exclude materials, energy and other intermediate inputs from the production function. Growing environmental pressures and resource prices suggest that this may be increasingly inappropriate. This paper explores the relationship between intermediate input intensity, productivity and national accounts using a panel dataset of manufacturing subsectors in the USA over 47 years. The first contribution is to identify sectoral production functions that incorporate intermediate inputs, while allowing for heterogeneity in both technology and productivity. The second contribution is that the paper finds a negative correlation between intermediate input intensity and total factor productivity (TFP)--sectors that are less intensive in their use of intermediate inputs have higher productivity. This finding is replicated at the firm level. We propose tentative hypotheses to explain this association, but testing and further disaggregation of intermediate inputs is left for further work. Further work could also explore more directly the relationship between material inputs and economic growth--given the high proportion of materials in intermediate inputs, the results in this paper are suggestive of further work on material efficiency. Depending upon the nature of the mechanism linking a reduction in intermediate input intensity to an increase in TFP, the implications could be significant. A third contribution is to suggest that an empirical bias in productivity, as measured in national accounts, may arise due to the exclusion of intermediate inputs. Current conventions of measuring productivity in national accounts may overstate the productivity of resource-intensive sectors relative to other sectors.

  20. Electrostatics Control Actin Filament Nucleation and Elongation Kinetics*

    Science.gov (United States)

    Crevenna, Alvaro H.; Naredi-Rainer, Nikolaus; Schönichen, André; Dzubiella, Joachim; Barber, Diane L.; Lamb, Don C.; Wedlich-Söldner, Roland

    2013-01-01

    The actin cytoskeleton is a central mediator of cellular morphogenesis, and rapid actin reorganization drives essential processes such as cell migration and cell division. Whereas several actin-binding proteins are known to be regulated by changes in intracellular pH, detailed information regarding the effect of pH on the actin dynamics itself is still lacking. Here, we combine bulk assays, total internal reflection fluorescence microscopy, fluorescence fluctuation spectroscopy techniques, and theory to comprehensively characterize the effect of pH on actin polymerization. We show that both nucleation and elongation are strongly enhanced at acidic pH, with a maximum close to the pI of actin. Monomer association rates are similarly affected by pH at both ends, although dissociation rates are differentially affected. This indicates that electrostatics control the diffusional encounter but not the dissociation rate, which is critical for the establishment of actin filament asymmetry. A generic model of protein-protein interaction, including electrostatics, explains the observed pH sensitivity as a consequence of charge repulsion. The observed pH effect on actin in vitro agrees with measurements of Listeria propulsion in pH-controlled cells. pH regulation should therefore be considered as a modulator of actin dynamics in a cellular environment. PMID:23486468

  1. Electrostatics control actin filament nucleation and elongation kinetics.

    Science.gov (United States)

    Crevenna, Alvaro H; Naredi-Rainer, Nikolaus; Schönichen, André; Dzubiella, Joachim; Barber, Diane L; Lamb, Don C; Wedlich-Söldner, Roland

    2013-04-26

    The actin cytoskeleton is a central mediator of cellular morphogenesis, and rapid actin reorganization drives essential processes such as cell migration and cell division. Whereas several actin-binding proteins are known to be regulated by changes in intracellular pH, detailed information regarding the effect of pH on the actin dynamics itself is still lacking. Here, we combine bulk assays, total internal reflection fluorescence microscopy, fluorescence fluctuation spectroscopy techniques, and theory to comprehensively characterize the effect of pH on actin polymerization. We show that both nucleation and elongation are strongly enhanced at acidic pH, with a maximum close to the pI of actin. Monomer association rates are similarly affected by pH at both ends, although dissociation rates are differentially affected. This indicates that electrostatics control the diffusional encounter but not the dissociation rate, which is critical for the establishment of actin filament asymmetry. A generic model of protein-protein interaction, including electrostatics, explains the observed pH sensitivity as a consequence of charge repulsion. The observed pH effect on actin in vitro agrees with measurements of Listeria propulsion in pH-controlled cells. pH regulation should therefore be considered as a modulator of actin dynamics in a cellular environment.

  2. Gestalt-binding of tropomyosin on actin during thin filament activation.

    Science.gov (United States)

    Lehman, William; Orzechowski, Marek; Li, Xiaochuan Edward; Fischer, Stefan; Raunser, Stefan

    2013-08-01

    Our thesis is that thin filament function can only be fully understood and muscle regulation then elucidated if atomic structures of the thin filament are available to reveal the positions of tropomyosin on actin in all physiological states. After all, it is tropomyosin influenced by troponin that regulates myosin-crossbridge cycling on actin and therefore controls contraction in all muscles. In addition, we maintain that a complete appreciation of thin filament activation also requires that the mechanical properties of tropomyosin itself are recognized and then related to the effect of myosin-association on actin. Taking the Gestalt-binding of tropomyosin into account, coupled with our electron microscopy structures and computational chemistry, we propose a comprehensive mechanism for tropomyosin regulatory movement over the actin filament surface that explains the cooperative muscle activation process. In fact, well-known point mutations of critical amino acids on the actin-tropomyosin binding interface disrupt Gestalt-binding and are associated with a number of inherited myopathies. Moreover, dysregulation of tropomyosin may also be a factor that interferes with the gatekeeping operation of non-muscle tropomyosin in the controlling interactions of a wide variety of cellular actin-binding proteins. The clinical relevance of Gestalt-binding is discussed in articles by the Marston and the Gunning groups in this special journal issue devoted to the impact of tropomyosin on biological systems.

  3. Differential mapping of the free barbed and pointed ends of actin filaments in cells.

    Science.gov (United States)

    Ofer, Noa; Abu Shah, Enas; Keren, Kinneret

    2014-06-01

    The actin cytoskeleton plays a pivotal role in many cellular processes. Detailed analysis of the architecture of cellular actin networks provides valuable insight into the dynamic self-organization underlying these processes. In particular, since most of the actin turnover occurs at the tips of actin filaments, it is insightful to map the distribution of filament ends. Here we report a method for differentially labeling the pointed and the barbed ends of actin filaments in cellular networks by permeabilizing cells, following a brief fixation, and introducing labeled actin monomers in the presence or absence of capping protein, respectively. This method quantitatively maps the distributions of free barbed ends and free pointed ends in adherent cells, providing information on the polarity of cytoskeletal structures and mapping active sites available for actin assembly or disassembly. We demonstrate the use of this method by mapping the distribution of actin filament ends in motile fish epithelial keratocytes and in several mammalian cell lines, and show that free barbed ends are enriched near the tip of protruding lamellipodia while free pointed ends concentrate toward the rear. © 2014 Wiley Periodicals, Inc.

  4. β-Helical architecture of cytoskeletal bactofilin filaments revealed by solid-state NMR

    Science.gov (United States)

    Vasa, Suresh; Lin, Lin; Shi, Chaowei; Habenstein, Birgit; Riedel, Dietmar; Kühn, Juliane; Thanbichler, Martin; Lange, Adam

    2015-01-01

    Bactofilins are a widespread class of bacterial filament-forming proteins, which serve as cytoskeletal scaffolds in various cellular pathways. They are characterized by a conserved architecture, featuring a central conserved domain (DUF583) that is flanked by variable terminal regions. Here, we present a detailed investigation of bactofilin filaments from Caulobacter crescentus by high-resolution solid-state NMR spectroscopy. De novo sequential resonance assignments were obtained for residues Ala39 to Phe137, spanning the conserved DUF583 domain. Analysis of the secondary chemical shifts shows that this core region adopts predominantly β-sheet secondary structure. Mutational studies of conserved hydrophobic residues located in the identified β-strand segments suggest that bactofilin folding and polymerization is mediated by an extensive and redundant network of hydrophobic interactions, consistent with the high intrinsic stability of bactofilin polymers. Transmission electron microscopy revealed a propensity of bactofilin to form filament bundles as well as sheet-like, 2D crystalline assemblies, which may represent the supramolecular arrangement of bactofilin in the native context. Based on the diffraction pattern of these 2D crystalline assemblies, scanning transmission electron microscopy measurements of the mass per length of BacA filaments, and the distribution of β-strand segments identified by solid-state NMR, we propose that the DUF583 domain adopts a β-helical architecture, in which 18 β-strand segments are arranged in six consecutive windings of a β-helix. PMID:25550503

  5. Dysfunctional mitochondria modulate cAMP-PKA signaling and filamentous and invasive growth of Saccharomyces cerevisiae.

    Science.gov (United States)

    Aun, Anu; Tamm, Tiina; Sedman, Juhan

    2013-02-01

    Mitochondrial metabolism is targeted by conserved signaling pathways that mediate external information to the cell. However, less is known about whether mitochondrial dysfunction interferes with signaling and thereby modulates the cellular response to environmental changes. In this study, we analyzed defective filamentous and invasive growth of the yeast Saccharomyces cerevisiae strains that have a dysfunctional mitochondrial genome (rho mutants). We found that the morphogenetic defect of rho mutants was caused by specific downregulation of FLO11, the adhesin essential for invasive and filamentous growth, and did not result from general metabolic changes brought about by interorganellar retrograde signaling. Transcription of FLO11 is known to be regulated by several signaling pathways, including the filamentous-growth-specific MAPK and cAMP-activated protein kinase A (cAMP-PKA) pathways. Our analysis showed that the filamentous-growth-specific MAPK pathway retained functionality in respiratory-deficient yeast cells. In contrast, the cAMP-PKA pathway was downregulated, explaining also various phenotypic traits observed in rho mutants. Thus, our results indicate that dysfunctional mitochondria modulate the output of the conserved cAMP-PKA signaling pathway.

  6. Arabidopsis actin-depolymerizing factor7 severs actin filaments and regulates actin cable turnover to promote normal pollen tube growth.

    Science.gov (United States)

    Zheng, Yiyan; Xie, Yurong; Jiang, Yuxiang; Qu, Xiaolu; Huang, Shanjin

    2013-09-01

    Actin filaments are often arranged into higher-order structures, such as the longitudinal actin cables that generate the reverse fountain cytoplasmic streaming pattern present in pollen tubes. While several actin binding proteins have been implicated in the generation of these cables, the mechanisms that regulate their dynamic turnover remain largely unknown. Here, we show that Arabidopsis thaliana actin-depolymerizing factor7 (ADF7) is required for turnover of longitudinal actin cables. In vitro biochemical analyses revealed that ADF7 is a typical ADF that prefers ADP-G-actin over ATP-G-actin. ADF7 inhibits nucleotide exchange on actin and severs filaments, but its filament severing and depolymerizing activities are less potent than those of the vegetative ADF1. ADF7 primarily decorates longitudinal actin cables in the shanks of pollen tubes. Consistent with this localization pattern, the severing frequency and depolymerization rate of filaments significantly decreased, while their maximum lifetime significantly increased, in adf7 pollen tube shanks. Furthermore, an ADF7-enhanced green fluorescent protein fusion with defective severing activity but normal G-actin binding activity could not complement adf7, providing compelling evidence that the severing activity of ADF7 is vital for its in vivo functions. These observations suggest that ADF7 evolved to promote turnover of longitudinal actin cables by severing actin filaments in pollen tubes.

  7. Filament Eruptions, Jets, and Space Weather

    Science.gov (United States)

    Moore, Ronald; Sterling, Alphonse; Robe, Nick; Falconer, David; Cirtain, Jonathan

    2013-01-01

    Previously, from chromospheric H alpha and coronal X-ray movies of the Sun's polar coronal holes, it was found that nearly all coronal jets (greater than 90%) are one or the other of two roughly equally common different kinds, different in how they erupt: standard jets and blowout jets (Yamauchi et al 2004, Apl, 605, 5ll: Moore et all 2010, Apj, 720, 757). Here, from inspection of SDO/AIA He II 304 A movies of 54 polar x-ray jets observed in Hinode/XRT movies, we report, as Moore et al (2010) anticipated, that (1) most standard x-ray jets (greater than 80%) show no ejected plasma that is cool enough (T is less than or approximately 10(exp 5K) to be seen in the He II 304 A movies; (2) nearly all blownout X-ray jets (greater than 90%) show obvious ejection of such cool plasma; (3) whereas when cool plasma is ejected in standard X-ray jets, it shows no lateral expansion, the cool plasma ejected in blowout X-ray jets shows strong lateral expansion; and (4) in many blowout X-ray jets, the cool plasma ejection displays the erupting-magnetic-rope form of clasic filament eruptions and is thereby seen to be a miniature filament eruption. The XRT movies also showed most blowout X-ray jets to be larger and brighter, and hence to apparently have more energy, than most standard X-ray jets. These observations (1) confirm the dichotomy of coronal jets, (2) agree with the Shibata model for standard jets, and (3) support the conclusion of Moore et al (2010) that in blowout jets the magnetic-arch base of the jet erupts in the manner of the much larger magnetic arcades in which the core field, the field rooted along the arcade's polarity inversion line, is sheared and twisted (sigmoid), often carries a cool-plasma filament, and erupts to blowout the arcade, producing a CME. From Hinode/SOT Ca II movies of the polar limb, Sterling et al (2010, ApJ, 714, L1) found that chromospheric Type-II spicules show a dichotomy of eruption dynamics similar to that found here for the cool

  8. The THMIS-MTR observation of a active region filament

    Science.gov (United States)

    Zong, W. G.; Tang, Y. H.; Fang, C.

    We present some THMIS-MTR observations of a active region filament on September 4, 2002. The full stokes parameters of the filament were obtained in Hα, CaII 8542 and FeI 6302. By use of the data with high spatial resolution(0.44" per pixel), we probed the fine structure of the filament and gave out the parameters at the barbs' endpoints, including intensity, velocity and longitudinal magnetic field. Comparing the quiescent filament which we have discussed before, we find that: 1)The velocities of the barbs' endpoints are much bigger in the active region filament, the values are more than one thousand meters per second. 2)The barbs' endpoints terminate at the low logitudinal magnetic field in the active region filament, too.

  9. A filament supported by different magnetic field configurations

    Science.gov (United States)

    Guo, Y.; Schmieder, B.; Démoulin, P.; Wiegelmann, T.; Aulanier, G.; Török, T.; Bommier, V.

    2011-08-01

    A nonlinear force-free magnetic field extrapolation of vector magnetogram data obtained by THEMIS/MTR on 2005 May 27 suggests the simultaneous existence of different magnetic configurations within one active region filament: one part of the filament is supported by field line dips within a flux rope, while the other part is located in dips within an arcade structure. Although the axial field chirality (dextral) and the magnetic helicity (negative) are the same along the whole filament, the chiralities of the filament barbs at different sections are opposite, i.e., right-bearing in the flux rope part and left-bearing in the arcade part. This argues against past suggestions that different barb chiralities imply different signs of helicity of the underlying magnetic field. This new finding about the chirality of filaments will be useful to associate eruptive filaments and magnetic cloud using the helicity parameter in the Space Weather Science.

  10. Reduced filamentation in high power semiconductor lasers

    DEFF Research Database (Denmark)

    Skovgaard, Peter M. W.; McInerney, John; O'Brien, Peter

    1999-01-01

    High brightness semiconductor lasers have applications in fields ranging from material processing to medicine. The main difficulty associated with high brightness is that high optical power densities cause damage to the laser facet and thus require large apertures. This, in turn, results in spatio......-temporal instabilities such as filamentation which degrades spatial coherence and brightness. We first evaluate performance of existing designs with a “top-hat” shaped transverse current density profile. The unstable nature of highly excited semiconductor material results in a run-away process where small modulations...

  11. The elastic modulus of isolated polytetrafluoroethylene filaments

    Directory of Open Access Journals (Sweden)

    Patrick Drawe

    2014-09-01

    Full Text Available We report vibrational Raman spectra of small extended perfluoro-n-alkanes (CnF2n+2 with n = 6, 8–10, 12–14 isolated in supersonic jet expansions and use wavenumbers of longitudinal acoustic vibrations to extrapolate the elastic modulus of cold, isolated polytetrafluoroethylene filaments. The derived value E = 209(10 GPa defines an upper limit for the elastic modulus of the perfectly crystalline, noninteracting polymer at low temperatures and serves as a benchmark for quantum chemical predictions.

  12. Filament winding cylinders. I - Process model

    Science.gov (United States)

    Lee, Soo-Yong; Springer, George S.

    1990-01-01

    A model was developed which describes the filament winding process of composite cylinders. The model relates the significant process variables such as winding speed, fiber tension, and applied temperature to the thermal, chemical and mechanical behavior of the composite cylinder and the mandrel. Based on the model, a user friendly code was written which can be used to calculate (1) the temperature in the cylinder and the mandrel, (2) the degree of cure and viscosity in the cylinder, (3) the fiber tensions and fiber positions, (4) the stresses and strains in the cylinder and in the mandrel, and (5) the void diameters in the cylinder.

  13. Welding. Performance Objectives. Intermediate Course.

    Science.gov (United States)

    Vincent, Kenneth

    Several intermediate performance objectives and corresponding criterion measures are listed for each of nine terminal objectives for an intermediate welding course. The materials were developed for a 36-week (3 hours daily) course designed to prepare the student for employment in the field of welding. Electric welding and specialized (TIG & MIG)…

  14. Multiple filamentation generated by focusing femtosecond laser with axicon.

    Science.gov (United States)

    Sun, Xiaodong; Gao, Hui; Zeng, Bin; Xu, Shengqi; Liu, Weiwei; Cheng, Ya; Xu, Zhizhan; Mu, Guoguang

    2012-03-01

    Multiple filamentation has been observed when focusing a femtosecond laser pulse into a methanol solution with an axicon. It is found that multiple long filaments are located on the central spot and ring structures of the quasi-Bessel beam created by the axicon. Since the quasi-Bessel profile is determined by the axicon properties, the axicon has been suggested as a simple optics to control multiple filaments. © 2012 Optical Society of America

  15. Observations of the Growth of an Active Region Filament

    Science.gov (United States)

    Yang, Bo

    2017-04-01

    We present observations of the growth of an active region filament caused by magnetic interactions among the filament and its adjacent superpenumbral filament (SF) and dark thread-like structures (T). Multistep reconnections are identified during the whole growing process. Magnetic flux convergence and cancellation occurring at the positive footpoint region of the filament is the first step reconnection, which resulted in the filament bifurcating into two sets of intertwined threads. One set anchored in situ, while the other set moved toward and interacted with the SF and part of T. This indicates the second step reconnection, which gave rise to the disappearance of the SF and the formation of a long thread-like structure that connects the far ends of the filament and T. The long thread-like structure further interacted with the T and then separated into two parts, representing the third step reconnection. Finally, another similar long thread-like structure, which intertwined with the fixed filament threads, appeared. Hαobservations show that this twisted structure is a longer sinistral filament. Based on the observed photospheric vector magnetograms, we performed a non-linear force-free field extrapolation to reconstruct the magnetic fields above the photosphere and found that the coronal magnetic field lines associated with the filament consists of two twisted flux ropes winding around each other. These results suggest that magnetic interactions among filaments and their adjacent SFs and T could lead to the growth of the filaments, and the filament is probably supported in a flux rope.

  16. Native bare zone assemblage nucleates myosin filament assembly.

    Science.gov (United States)

    Niederman, R; Peters, L K

    1982-11-15

    Native myosin filaments from rabbit psoas muscle are always 1.5 micrometer long. The regulated assembly of these filaments is generally considered to occur by an initial antiparallel and subsequent parallel aggregation of identical myosin subunits. In this schema myosin filament length is controlled by either a self-assembly or a Vernier process. We present evidence which refines these ideas. Namely, that the intact myosin bare zone assemblage nucleates myosin filament assembly. This suggestion is based on the following experimental evidence. (1) A native bare zone assemblage about 0.3 micrometer long can be formed by dialysis of native myosin filaments to either a pH 8 or a 0.2 M-KCl solution. (2) Upon dialysis back to 0.1 M-KCl, bare zone assemblages and distal myosin molecules recombine to form 1.5 micrometer long bipolar filaments. (3) The bare zone assemblage can be separated from the distal myosin molecules by column chromatography in 0.2 M-KCl. Upon dialysis of the fractionated subsets back to 0.1 M-KCl, the bare zone assemblage retains its length of about 0.3 micrometer. However, the distal molecules reassemble to form filaments about 5 micrometers long. (4) Filaments are formed from mixes of the isolated subsets. The lengths of these filaments vary with the amount of distal myosin present. (5) When native filaments, isolated bare zone assemblages or distal myosin molecules are moved sequentially to 0.6 M-KCl and then to 0.1 M-KCl, the final filament lengths are all about 5 micrometers. The capacity of the bare zone assemblage to nucleate filament assembly may be due to the bare zone myosin molecules, the associated M band components or both.

  17. Dynamic Regulation of Sarcomeric Actin Filaments in Striated Muscle

    OpenAIRE

    Ono, Shoichiro

    2010-01-01

    In striated muscle, the actin cytoskeleton is differentiated into myofibrils. Actin and myosin filaments are organized in sarcomeres and specialized for producing contractile forces. Regular arrangement of actin filaments with uniform length and polarity is critical for the contractile function. However, the mechanisms of assembly and maintenance of sarcomeric actin filaments in striated muscle are not completely understood. Live imaging of actin in striated muscle has revealed that actin sub...

  18. Modeling Vertical Plasma Flows in Solar Filament Barbs

    Science.gov (United States)

    Litvinenko, Y.

    2003-12-01

    Speeds of observed flows in quiescent solar filaments are typically much less than the local Alfvén speed. This is why the flows in filament barbs can be modeled by perturbing a local magnetostatic solution describing the balance between the Lorentz force, gravity, and gas pressure in a barb. Similarly, large-scale filament flows can be treated as adiabatically slow deformations of a force-free magnetic equilibrium that describes the global structure of a filament. This approach reconciles current theoretical models with the puzzling observational result that some of the flows appear to be neither aligned with the magnetic field nor controlled by gravity.

  19. Filament shape versus coronal potential magnetic field structure

    Science.gov (United States)

    Filippov, B.

    2016-01-01

    Solar filament shape in projection on disc depends on the structure of the coronal magnetic field. We calculate the position of polarity inversion lines (PILs) of coronal potential magnetic field at different heights above the photosphere, which compose the magnetic neutral surface, and compare with them the distribution of the filament material in Hα chromospheric images. We found that the most of the filament material is enclosed between two PILs, one at a lower height close to the chromosphere and one at a higher level, which can be considered as a height of the filament spine. Observations of the same filament on the limb by the Solar Terrestrial Relations Observatory spacecraft confirm that the height of the spine is really very close to the value obtained from the PIL and filament border matching. Such matching can be used for filament height estimations in on-disc observations. Filament barbs are housed within protruding sections of the low-level PIL. On the base of simple model, we show that the similarity of the neutral surfaces in potential and non-potential fields with the same sub-photospheric sources is the reason for the found tendency for the filament material to gather near the potential-field neutral surface.

  20. Statistical Study of the Magnetic Field Orientation in Solar Filaments

    Science.gov (United States)

    Hanaoka, Yoichiro; Sakurai, Takashi

    2017-12-01

    We have carried out a statistical study of the average orientation of the magnetic field in solar filaments with respect to their axes for more than 400 samples, based on data taken with daily full-Sun, full-Stokes spectropolarimetric observations using the He I 1083.0 nm line. The major part of the samples are the filaments in the quiet areas, but those in the active areas are included as well. The average orientation of the magnetic field in filaments shows a systematic property depending on the hemisphere; the direction of the magnetic field in filaments in the northern (southern) hemisphere mostly deviates clockwise (counterclockwise) from their axes, which run along the magnetic polarity inversion line. The deviation angles of the magnetic field from the axes are concentrated between 10° and 30°. This hemispheric pattern is consistent with that revealed for chirality of filament barbs, filament channels, and for other solar features found to possess chirality. For some filaments, it was confirmed that their magnetic field direction is locally parallel to their structure seen in Hα images. Our results for the first time confirmed this hemispheric pattern with the direct observation of the magnetic field in filaments. Interestingly, the filaments which show the opposite magnetic field deviation to the hemispheric pattern, are in many cases found above the polarity inversion line whose ambient photospheric magnetic field has the polarity alignment being opposite to that of active regions following the Hale–Nicholson law.

  1. Spatial evolution of laser filaments in turbulent air

    Science.gov (United States)

    Zeng, Tao; Zhu, Shiping; Zhou, Shengling; He, Yan

    2018-04-01

    In this study, the spatial evolution properties of laser filament clusters in turbulent air were evaluated using numerical simulations. Various statistical parameters were calculated, such as the percolation probability, filling factor, and average cluster size. The results indicate that turbulence-induced multi-filamentation can be described as a new phase transition universality class. In addition, during this process, the relationship between the average cluster size and filling factor could be fit by a power function. Our results are valuable for applications involving filamentation that can be influenced by the geometrical features of multiple filaments.

  2. Numerical simulation of laser filamentation in underdense plasma

    International Nuclear Information System (INIS)

    Yu Lichun; Chen Zhihua; Tu Qinfen

    2000-01-01

    Developing process of filamentation and effect of characteristic parameters in underdense plasma have been studied using numerical simulation method. Production and development of two-dimensional cylinder filamentation instability were presented clearly. The results indicate incidence laser intensity and plasma background density are important factors affecting convergent intensity. At the same time, it was showed that different laser wavelength or different electron background density could affect filamentation process. The results are consistent with theory and experiments of alien reports. It can provide reference for restraining filamentation

  3. MATERIAL SUPPLY AND MAGNETIC CONFIGURATION OF AN ACTIVE REGION FILAMENT

    Energy Technology Data Exchange (ETDEWEB)

    Zou, P.; Fang, C.; Chen, P. F.; Yang, K.; Hao, Q. [School of Astronomy and Space Science, Nanjing University, Nanjing 210023 (China); Cao, Wenda, E-mail: fangc@nju.edu.cn [Big Bear Solar Observatory, New Jersey Institute of Technology, 40386 North Shore Lane, Big Bear City, CA 92314 (United States)

    2016-11-10

    It is important to study the fine structures of solar filaments with high-resolution observations, since it can help us understand the magnetic and thermal structures of the filaments and their dynamics. In this paper, we study a newly formed filament located inside the active region NOAA 11762, which was observed by the 1.6 m New Solar Telescope at Big Bear Solar Observatory from 16:40:19 UT to 17:07:58 UT on 2013 June 5. As revealed by the H α filtergrams, cool material is seen to be injected into the filament spine with a speed of 5–10 km s{sup -1}. At the source of the injection, brightenings are identified in the chromosphere, which are accompanied by magnetic cancellation in the photosphere, implying the importance of magnetic reconnection in replenishing the filament with plasmas from the lower atmosphere. Counter-streamings are detected near one endpoint of the filament, with the plane-of-the-sky speed being 7–9 km s{sup -1} in the H α red-wing filtergrams and 9–25 km s{sup -1} in the blue-wing filtergrams. The observations are indicative that this active region filament is supported by a sheared arcade without magnetic dips, and the counter-streamings are due to unidirectional flows with alternative directions, rather than due to the longitudinal oscillations of filament threads as in many other filaments.

  4. Human Rad51 filaments on double- and single-stranded DNA: correlating regular and irregular forms with recombination function.

    NARCIS (Netherlands)

    D. Ristic (Dejan); M. Modesti (Mauro); T. van der Heijden (Thijn); J. Noort (John); C. Dekker (Cees); R. Kanaar (Roland); C. Wyman (Claire)

    2005-01-01

    textabstractRecombinase proteins assembled into helical filaments on DNA are believed to be the catalytic core of homologous recombination. The assembly, disassembly and dynamic rearrangements of this structure must drive the DNA strand exchange reactions of homologous recombination. The sensitivity

  5. Assembly of intermediates for rapid membrane fusion.

    Science.gov (United States)

    Harner, Max; Wickner, William

    2018-01-26

    Membrane fusion is essential for intracellular protein sorting, cell growth, hormone secretion, and neurotransmission. Rapid membrane fusion requires tethering and Sec1-Munc18 (SM) function to catalyze R-, Qa-, Qb-, and Qc-SNARE complex assembly in trans , as well as SNARE engagement by the SNARE-binding chaperone Sec17/αSNAP. The hexameric vacuolar HOPS ( ho motypic fusion and vacuole p rotein s orting) complex in the yeast Saccharomyces cerevisiae tethers membranes through its affinities for the membrane Rab GTPase Ypt7. HOPS also has specific affinities for the vacuolar SNAREs and catalyzes SNARE complex assembly, but the order of their assembly into a 4-SNARE complex is unclear. We now report defined assembly intermediates on the path to membrane fusion. We found that a prefusion intermediate will assemble with HOPS and the R, Qa, and Qc SNAREs, and that this assembly undergoes rapid fusion upon addition of Qb and Sec17. HOPS-tethered membranes and all four vacuolar SNAREs formed a complex that underwent an even more dramatic burst of fusion upon Sec17p addition. These findings provide initial insights into an ordered fusion pathway consisting of the following intermediates and events: 1) Rab- and HOPS-tethered membranes, 2) a HOPS:R:Qa:Qc trans -complex, 3) a HOPS:4-SNARE trans -complex, 4) an engagement with Sec17, and 5) the rapid lipid rearrangements during fusion. In conclusion, our results indicate that the R:Qa:Qc complex forms in the context of membrane, Ypt7, HOPS, and trans -SNARE assembly and serves as a functional intermediate for rapid fusion after addition of the Qb-SNARE and Sec17 proteins. © 2018 by The American Society for Biochemistry and Molecular Biology, Inc.

  6. A Synchrotron-Based Hydroxyl Radical Footprinting Analysis of Amyloid Fibrils and Prefibrillar Intermediates with Residue-Specific Resolution

    Energy Technology Data Exchange (ETDEWEB)

    Klinger, Alexandra L. [Univ. of Pennsylvania, Philadelphia, PA (United States); Kiselar, Janna [Case Western Reserve Univ., Cleveland, OH (United States); Ilchenko, Serguei [Case Western Reserve Univ., Cleveland, OH (United States); Komatsu, Hiroaki [Univ. of Pennsylvania, Philadelphia, PA (United States); Chance, Mark R. [Case Western Reserve Univ., Cleveland, OH (United States); Axelsen, Paul H. [Univ. of Pennsylvania, Philadelphia, PA (United States)

    2014-11-09

    The structural models of the fibrils formed by the 40-residue amyloid-β (Aβ40) peptide in Alzheimer’s disease typically consist of linear polypeptide segments, oriented approximately perpendicular to the long axis of the fibril, and joined together as parallel in-register β-sheets to form filaments. However, various models differ in the number of filaments that run the length of a fibril, and in the topological arrangement of these filaments. In addition to questions about the structure of Aβ40 monomers in fibrils, there are important unanswered questions about their structure in prefibrillar intermediates, which are of interest because they may represent the most neurotoxic form of Aβ40. To assess different models of fibril structure and to gain insight into the structure of prefibrillar intermediates, the relative solvent accessibility of amino acid residue side chains in fibrillar and prefibrillar Aβ40 preparations was characterized in solution by hydroxyl radical footprinting and structural mass spectrometry. A key to the application of this technology was the development of hydroxyl radical reactivity measures for individual side chains of Aβ40. When we combined mass-per-length measurements performed by dark-field electron microscopy, we determined that the results of our study were consistent with the core filament structure represented by two- and three-filament solid state nuclear magnetic resonance-based models of the Aβ40 fibril (such as 2LMN, 2LMO, 2LMP, and 2LMQ), with minor refinements, but they are inconsistent with the more recently proposed 2M4J model. Our results also demonstrate that individual Aβ40 fibrils exhibit structural heterogeneity or polymorphism, where regions of two-filament structure alternate with regions of three-filament structure. The footprinting approach utilized in this study will be valuable for characterizing various fibrillar and nonfibrillar forms of the Aβ peptide.

  7. Homologous Transcription Factors DUX4 and DUX4c Associate with Cytoplasmic Proteins during Muscle Differentiation.

    Directory of Open Access Journals (Sweden)

    Eugénie Ansseau

    Full Text Available Hundreds of double homeobox (DUX genes map within 3.3-kb repeated elements dispersed in the human genome and encode DNA-binding proteins. Among these, we identified DUX4, a potent transcription factor that causes facioscapulohumeral muscular dystrophy (FSHD. In the present study, we performed yeast two-hybrid screens and protein co-purifications with HaloTag-DUX fusions or GST-DUX4 pull-down to identify protein partners of DUX4, DUX4c (which is identical to DUX4 except for the end of the carboxyl terminal domain and DUX1 (which is limited to the double homeodomain. Unexpectedly, we identified and validated (by co-immunoprecipitation, GST pull-down, co-immunofluorescence and in situ Proximal Ligation Assay the interaction of DUX4, DUX4c and DUX1 with type III intermediate filament protein desmin in the cytoplasm and at the nuclear periphery. Desmin filaments link adjacent sarcomere at the Z-discs, connect them to sarcolemma proteins and interact with mitochondria. These intermediate filament also contact the nuclear lamina and contribute to positioning of the nuclei. Another Z-disc protein, LMCD1 that contains a LIM domain was also validated as a DUX4 partner. The functionality of DUX4 or DUX4c interactions with cytoplasmic proteins is underscored by the cytoplasmic detection of DUX4/DUX4c upon myoblast fusion. In addition, we identified and validated (by co-immunoprecipitation, co-immunofluorescence and in situ Proximal Ligation Assay as DUX4/4c partners several RNA-binding proteins such as C1QBP, SRSF9, RBM3, FUS/TLS and SFPQ that are involved in mRNA splicing and translation. FUS and SFPQ are nuclear proteins, however their cytoplasmic translocation was reported in neuronal cells where they associated with ribonucleoparticles (RNPs. Several other validated or identified DUX4/DUX4c partners are also contained in mRNP granules, and the co-localizations with cytoplasmic DAPI-positive spots is in keeping with such an association. Large muscle RNPs

  8. Electromechanical vortex filaments during cardiac fibrillation

    Science.gov (United States)

    Christoph, J.; Chebbok, M.; Richter, C.; Schröder-Schetelig, J.; Bittihn, P.; Stein, S.; Uzelac, I.; Fenton, F. H.; Hasenfuß, G.; Gilmour, R. F., Jr.; Luther, S.

    2018-03-01

    The self-organized dynamics of vortex-like rotating waves, which are also known as scroll waves, are the basis of the formation of complex spatiotemporal patterns in many excitable chemical and biological systems. In the heart, filament-like phase singularities that are associated with three-dimensional scroll waves are considered to be the organizing centres of life-threatening cardiac arrhythmias. The mechanisms that underlie the onset, maintenance and control of electromechanical turbulence in the heart are inherently three-dimensional phenomena. However, it has not previously been possible to visualize the three-dimensional spatiotemporal dynamics of scroll waves inside cardiac tissues. Here we show that three-dimensional mechanical scroll waves and filament-like phase singularities can be observed deep inside the contracting heart wall using high-resolution four-dimensional ultrasound-based strain imaging. We found that mechanical phase singularities co-exist with electrical phase singularities during cardiac fibrillation. We investigated the dynamics of electrical and mechanical phase singularities by simultaneously measuring the membrane potential, intracellular calcium concentration and mechanical contractions of the heart. We show that cardiac fibrillation can be characterized using the three-dimensional spatiotemporal dynamics of mechanical phase singularities, which arise inside the fibrillating contracting ventricular wall. We demonstrate that electrical and mechanical phase singularities show complex interactions and we characterize their dynamics in terms of trajectories, topological charge and lifetime. We anticipate that our findings will provide novel perspectives for non-invasive diagnostic imaging and therapeutic applications.

  9. Morgellons disease: a filamentous borrelial dermatitis

    Directory of Open Access Journals (Sweden)

    Middelveen MJ

    2016-10-01

    Full Text Available Marianne J Middelveen, Raphael B Stricker International Lyme and Associated Diseases Society, Bethesda, MD, USA Abstract: Morgellons disease (MD is a dermopathy characterized by multicolored filaments that lie under, are embedded in, or project from skin. Although MD was initially considered to be a delusional disorder, recent studies have demonstrated that the dermopathy is associated with tickborne infection, that the filaments are composed of keratin and collagen, and that they result from proliferation of keratinocytes and fibroblasts in epithelial tissue. Culture, histopathological and molecular evidence of spirochetal infection associated with MD has been presented in several published studies using a variety of techniques. Spirochetes genetically identified as Borrelia burgdorferi sensu stricto predominate as the infective agent in most of the Morgellons skin specimens studied so far. Other species of Borrelia including Borrelia garinii, Borrelia miyamotoi, and Borrelia hermsii have also been detected in skin specimens taken from MD patients. The optimal treatment for MD remains to be determined. Keywords: Morgellons disease, dermatitis, Lyme disease, Borrelia burgdorferi, spirochetes

  10. A monoclonal antibody marker for the exclusion-zone filaments of Trypanosoma brucei.

    Science.gov (United States)

    Bonhivers, Mélanie; Landrein, Nicolas; Decossas, Marion; Robinson, Derrick R

    2008-07-10

    Trypanosoma brucei is a haemoflagellate pathogen of man, wild animals and domesticated livestock in central and southern Africa. In all life cycle stages this parasite has a single mitochondrion that contains a uniquely organised genome that is condensed into a flat disk-like structure called the kinetoplast. The kinetoplast is essential for insect form procyclic cells and therefore is a potential drug target. The kinetoplast is unique in nature because it consists of novel structural proteins and thousands of circular, interlocking, DNA molecules (kDNA). Secondly, kDNA replication is critically timed to coincide with nuclear S phase and new flagellum biogenesis. Thirdly, the kinetoplast is physically attached to the flagellum basal bodies via a structure called the tripartite attachment complex (TAC). The TAC consists of unilateral filaments (within the mitochondrion matrix), differentiated mitochondrial membranes and exclusion-zone filaments that extend from the distal end of the basal bodies. To date only one protein, p166, has been identified to be a component of the TAC. In the work presented here we provide data based on a novel EM technique developed to label and characterise cytoskeleton structures in permeabilised cells without extraction of mitochondrion membranes. We use this protocol to provide data on a new monoclonal antibody reagent (Mab 22) and illustrate the precise localisation of basal body-mitochondrial linker proteins. Mab 22 binds to these linker proteins (exclusion-zone filaments) and provides a new tool for the characterisation of cytoskeleton mediated kinetoplast segregation. The antigen(s) recognised by Mab 22 are cytoskeletal, insensitive to extraction by high concentrations of non-ionic detergent, extend from the proximal region of basal bodies and bind to the outer mitochondrial membrane. This protein(s) is the first component of the TAC exclusion-zone fibres to be identified. Mab 22 will therefore be important in characterising TAC

  11. A monoclonal antibody marker for the exclusion-zone filaments of Trypanosoma brucei

    Directory of Open Access Journals (Sweden)

    Decossas Marion

    2008-07-01

    Full Text Available Abstract Background Trypanosoma brucei is a haemoflagellate pathogen of man, wild animals and domesticated livestock in central and southern Africa. In all life cycle stages this parasite has a single mitochondrion that contains a uniquely organised genome that is condensed into a flat disk-like structure called the kinetoplast. The kinetoplast is essential for insect form procyclic cells and therefore is a potential drug target. The kinetoplast is unique in nature because it consists of novel structural proteins and thousands of circular, interlocking, DNA molecules (kDNA. Secondly, kDNA replication is critically timed to coincide with nuclear S phase and new flagellum biogenesis. Thirdly, the kinetoplast is physically attached to the flagellum basal bodies via a structure called the tripartite attachment complex (TAC. The TAC consists of unilateral filaments (within the mitochondrion matrix, differentiated mitochondrial membranes and exclusion-zone filaments that extend from the distal end of the basal bodies. To date only one protein, p166, has been identified to be a component of the TAC. Results In the work presented here we provide data based on a novel EM technique developed to label and characterise cytoskeleton structures in permeabilised cells without extraction of mitochondrion membranes. We use this protocol to provide data on a new monoclonal antibody reagent (Mab 22 and illustrate the precise localisation of basal body-mitochondrial linker proteins. Mab 22 binds to these linker proteins (exclusion-zone filaments and provides a new tool for the characterisation of cytoskeleton mediated kinetoplast segregation. Conclusion The antigen(s recognised by Mab 22 are cytoskeletal, insensitive to extraction by high concentrations of non-ionic detergent, extend from the proximal region of basal bodies and bind to the outer mitochondrial membrane. This protein(s is the first component of the TAC exclusion-zone fibres to be identified. Mab 22

  12. Hemidesmosomal linker proteins regulate cell motility, invasion and tumorigenicity in oral squamous cell carcinoma derived cells.

    Science.gov (United States)

    Chaudhari, Pratik Rajeev; Charles, Silvania Emlit; D'Souza, Zinia Charlotte; Vaidya, Milind Murlidhar

    2017-11-15

    BPAG1e and Plectin are hemidesmosomal linker proteins which anchor intermediate filament proteins to the cell surface through β4 integrin. Recent reports indicate that these proteins play a role in various cellular processes apart from their known anchoring function. However, the available literature is inconsistent. Further, the previous study from our laboratory suggested that Keratin8/18 pair promotes cell motility and tumor progression by deregulating β4 integrin signaling in oral squamous cell carcinoma (OSCC) derived cells. Based on these findings, we hypothesized that linker proteins may have a role in neoplastic progression of OSCC. Downregulation of hemidesmosomal linker proteins in OSCC derived cells resulted in reduced cell migration accompanied by alterations in actin organization. Further, decreased MMP9 activity led to reduced cell invasion in linker proteins knockdown cells. Moreover, loss of these proteins resulted in reduced tumorigenic potential. SWATH analysis demonstrated upregulation of N-Myc downstream regulated gene 1 (NDRG1) in linker proteins downregulated cells as compared to vector control cells. Further, the defects in phenotype upon linker proteins ablation were rescued upon loss of NDRG1 in linker proteins knockdown background. These data together indicate that hemidesmosomal linker proteins regulate cell motility, invasion and tumorigenicity possibly through NDRG1 in OSCC derived cells. Copyright © 2017 Elsevier Inc. All rights reserved.

  13. Malaria parasites form filamentous cell-to-cell connections during reproduction in the mosquito midgut.

    Science.gov (United States)

    Rupp, Ingrid; Sologub, Ludmilla; Williamson, Kim C; Scheuermayer, Matthias; Reininger, Luc; Doerig, Christian; Eksi, Saliha; Kombila, Davy U; Frank, Matthias; Pradel, Gabriele

    2011-04-01

    Physical contact is important for the interaction between animal cells, but it can represent a major challenge for protists like malaria parasites. Recently, novel filamentous cell-cell contacts have been identified in different types of eukaryotic cells and termed nanotubes due to their morphological appearance. Nanotubes represent small dynamic membranous extensions that consist of F-actin and are considered an ancient feature evolved by eukaryotic cells to establish contact for communication. We here describe similar tubular structures in the malaria pathogen Plasmodium falciparum, which emerge from the surfaces of the forming gametes upon gametocyte activation in the mosquito midgut. The filaments can exhibit a length of > 100 μm and contain the F-actin isoform actin 2. They actively form within a few minutes after gametocyte activation and persist until the zygote transforms into the ookinete. The filaments originate from the parasite plasma membrane, are close ended and express adhesion proteins on their surfaces that are typically found in gametes, like Pfs230, Pfs48/45 or Pfs25, but not the zygote surface protein Pfs28. We show that these tubular structures represent long-distance cell-to-cell connections between sexual stage parasites and demonstrate that they meet the characteristics of nanotubes. We propose that malaria parasites utilize these adhesive "nanotubes" in order to facilitate intercellular contact between gametes during reproduction in the mosquito midgut.

  14. Serological and genetic characterisation of bovine respiratory syncytial virus (BRSV) indicates that Danish isolates belong to the intermediate subgroup: no evidence of a selective effect on the variability of G protein nucleotide sequence by prior cell culture adaption and passages in cell culture

    DEFF Research Database (Denmark)

    Larsen, Lars Erik; Uttenthal, Åse; Arctander, P.

    1998-01-01

    , respectively. Phylogenetic analysis showed that the Danish isolates formed three lineages within a separate branch of the phylogenetic tree. Nevertheless, the Danish isolates were closely related to the 220-69Bel isolate, the prototype of the intermediate antigenic subgroup. The sequencing of the extracellular...... part of the G gene of additional 11 field BRSV viruses, processed directly from lung samples without prior adaption to cell culture growth. revealed sequence variabilities in the range obtained with the propagated virus. In addition, several passages in cell culture and in calves had no major impact...... on the nucleotide sequence of the G protein. These findings indicated that the previously established variabilities of the G protein of RS virus isolates were not attributable to mutations induced during the propagation of the virus. The reactivity of the Danish isolates with G protein-specific MAbs were similar...

  15. A catalytic oligomeric motor that walks along a filament track

    Science.gov (United States)

    Huang, Mu-Jie; Kapral, Raymond

    2015-06-01

    Most biological motors in the cell execute chemically powered conformational changes as they walk on biopolymer filaments in order to carry out directed transport functions. Synthetic motors that operate in a similar manner are being studied since they have the potential to perform similar tasks in a variety of applications. In this paper, a synthetic nanomotor that moves along a filament track, without invoking motor conformational changes, is constructed and its properties are studied in detail. The motor is an oligomer comprising three linked beads with specific binding properties. The filament track is a stiff polymer chain, also described by a linear chain of linked coarse-grained molecular groups modeled as beads. Reactions on the filament that are catalyzed by a motor bead and use fuel in the environment, in conjunction within the binding affinities of the motor beads to the filament beads, lead to directed motion. The system operates out of equilibrium due to the state of the filament and supply of fuel. The motor, filament, and surrounding medium are all described at microscopic level that permits a full analysis of the motor motion. A stochastic model that captures the main trends seen in the simulations is also presented. The results of this study point to some of the key features that could be used to construct nanomotors that undergo biased walks powered by chemical reactions on filaments.

  16. Application of digital holography to filament size analysis

    NARCIS (Netherlands)

    Semin, N.V.; Poelma, C.; Drost, S.; Westerweel, J.

    2010-01-01

    The potential of in-line digital holography to locate and measure the size and position of filaments, i.e. thin wire-like objects, distributed throughout a thick volume has been investigated. In this paper two approaches are introduced to study filaments of varying diameter. (1) It is shown

  17. Fully filamentized HTS coated conductor via striation and selective electroplating

    Energy Technology Data Exchange (ETDEWEB)

    Kesgin, Ibrahim; Majkic, Goran [Department of Mechanical Engineering and Texas Center for Superconductivity, University of Houston, Houston, TX 77204 (United States); Selvamanickam, Venkat, E-mail: selva@uh.edu [Department of Mechanical Engineering and Texas Center for Superconductivity, University of Houston, Houston, TX 77204 (United States)

    2013-03-15

    Highlights: ► Fully-filamentized coated conductor with 13-fold reduction in ac losses. ► Selective electroplating for filamentization of thick copper stabilizer. ► A twofold decrease in ac loss by filamentization of copper stabilizer. ► Absence of appreciable coupling loss contribution from electroplating. -- Abstract: A simple, cost-effective method involving top-down mechanical scribing, oxidation and bottom-up electroplating has been successfully developed to fabricate fully filamentized HTS coated conductors. The copper stabilizer layer is selectively electroplated on the superconducting filaments while the striations remain copper-free due to the formation of a resistive oxide layer in between filaments by oxidation of the striated grooves at elevated temperature in oxygen atmosphere. Magnetization AC loss measurements, performed in a frequency range of 45–500 Hz at 77 K, confirmed the expected N-fold reduction in AC loss of the filamentized tapes with no significant degradation in critical current beyond that due to the material removal from the striations (N – number of filaments). A considerable reduction in coupling AC loss was observed after high temperature annealing/oxidation of the striated tapes. Furthermore, a significant reduction in eddy current loss was achieved with selective copper electroplating, as evidenced by analyzing the field and frequency dependence of magnetization AC loss, as well as by comparing the AC loss performance of striated samples to that of non-striated samples after electroplating of copper stabilizer.

  18. Fully filamentized HTS coated conductor via striation and selective electroplating

    International Nuclear Information System (INIS)

    Kesgin, Ibrahim; Majkic, Goran; Selvamanickam, Venkat

    2013-01-01

    Highlights: ► Fully-filamentized coated conductor with 13-fold reduction in ac losses. ► Selective electroplating for filamentization of thick copper stabilizer. ► A twofold decrease in ac loss by filamentization of copper stabilizer. ► Absence of appreciable coupling loss contribution from electroplating. -- Abstract: A simple, cost-effective method involving top-down mechanical scribing, oxidation and bottom-up electroplating has been successfully developed to fabricate fully filamentized HTS coated conductors. The copper stabilizer layer is selectively electroplated on the superconducting filaments while the striations remain copper-free due to the formation of a resistive oxide layer in between filaments by oxidation of the striated grooves at elevated temperature in oxygen atmosphere. Magnetization AC loss measurements, performed in a frequency range of 45–500 Hz at 77 K, confirmed the expected N-fold reduction in AC loss of the filamentized tapes with no significant degradation in critical current beyond that due to the material removal from the striations (N – number of filaments). A considerable reduction in coupling AC loss was observed after high temperature annealing/oxidation of the striated tapes. Furthermore, a significant reduction in eddy current loss was achieved with selective copper electroplating, as evidenced by analyzing the field and frequency dependence of magnetization AC loss, as well as by comparing the AC loss performance of striated samples to that of non-striated samples after electroplating of copper stabilizer

  19. Fossil evidence for spin alignment of SDSS galaxies in filaments

    NARCIS (Netherlands)

    Jones, Bernard J.T.; Weygaert, Rien van de; Arag´on-Calvo, Miguel A.

    2010-01-01

    We search for and find fossil evidence that the distribution of the spin axes of galaxies in cosmic web filaments relative to their host filaments are not randomly distributed. This would indicate that the action of large scale tidal torques effected the alignments of galaxies located in cosmic

  20. Method for simultaneously coating a plurality of filaments

    Science.gov (United States)

    Miller, P.A.; Pochan, P.D.; Siegal, M.P.; Dominguez, F.

    1995-07-11

    Methods and apparatuses are disclosed for coating materials, and the products and compositions produced thereby. Substances, such as diamond or diamond-like carbon, are deposited onto materials, such as a filament or a plurality of filaments simultaneously, using one or more cylindrical, inductively coupled, resonator plasma reactors. 3 figs.

  1. Cellulase activity of filamentous fungi induced by rice husk | Oliveros ...

    African Journals Online (AJOL)

    Cellulase activity of filamentous fungi induced by rice husk. DF Oliveros, N Guarnizo, EM Perea, WM Arango. Abstract. The objective of this study was to determine the potential of different filamentous fungi to degrade cellulose in rice husk pre-treated with steam explosion or alkaline hydrolysis. A preliminary test performed ...

  2. Calibration and Temperature Profile of a Tungsten Filament Lamp

    Science.gov (United States)

    de Izarra, Charles; Gitton, Jean-Michel

    2010-01-01

    The goal of this work proposed for undergraduate students and teachers is the calibration of a tungsten filament lamp from electric measurements that are both simple and precise, allowing to determine the temperature of tungsten filament as a function of the current intensity. This calibration procedure was first applied to a conventional filament…

  3. Design and Optimization of Filament Wound Composite Pressure Vessels

    NARCIS (Netherlands)

    Zu, L.

    2012-01-01

    One of the most important issues for the design of filament-wound pressure vessels reflects on the determination of the most efficient meridian profiles and related fiber architectures, leading to optimal structural performance. To better understand the design and optimization of filament-wound

  4. THE APPARATUS FOR ALIGNMENT OF THE PHOTOMETRIC LAMP FILAMENT

    Directory of Open Access Journals (Sweden)

    V. A. Dlugunovich

    2015-01-01

    Full Text Available During photometric measurements involving the use of photometric lamps it is necessary that the filament of lamp takes a strictly predetermined position with respect to the photodetector and the optical axis of the photometric setup. The errors in positioning of alignment filament with respect to the optical axis of the measuring system lead to increase the uncertainty of measurement of the photometric characteristics of the light sources. A typical method for alignment of filament of photometric lamps is based on the use a diopter tubes (telescopes. Using this method, the mounting of filament to the required position is carried out by successive approximations, which requires special concentration and a lot of time. The aim of this work is to develop an apparatus for alignment which allows simultaneous alignment of the filament of lamps in two mutually perpendicular planes. The method and apparatus for alignment of the photometric lamp filament during measurements of the photometric characteristics of light sources based on two digital video cameras is described in this paper. The apparatus allows to simultaneously displaying the image of lamps filament on the computer screen in two mutually perpendicular planes. The apparatus eliminates a large number of functional units requiring elementwise alignment and reduces the time required to carry out the alignment. The apparatus also provides the imaging of lamps filament with opaque coated on the bulb. The apparatus is used at the National standard of light intensity and illuminance units of the Republic of Belarus. 

  5. The role of Bgl2p in the transition to filamentous cells during biofilm formation by Candida albicans.

    Science.gov (United States)

    Chen, Xinyue; Zhang, Ruoyu; Takada, Ayako; Iwatani, Shun; Oka, Chiemi; Kitamoto, Toshitaka; Kajiwara, Susumu

    2017-02-01

    The fungal pathogen Candida albicans undergoes a transition from yeast cells to filamentous cells that is related to its pathogenicity. The complex multicellular processes involved in biofilm formation by this fungus also include this transition. In this work, we investigated the morphological role of the Bgl2 protein (Bgl2p) in the transition to filamentous cells during biofilm formation by C. albicans. Bgl2p has been identified as a β-1, 3-glucosyltransferase, and transcription of the CaBGL2 gene is upregulated during biofilm formation. We used scanning electron microscopy to observe the microstructure of a bgl2 null mutant during biofilm formation and found a delay in the transition to filamentous cells in the premature phase (24 hours) of biofilm formation. Deletion of the CaBGL2 gene led to a decrease in the expression of CPH2 and TEC1, which encode transcription factors required for the transition to the filamentous form. These findings indicate that Bgl2p plays a role in the transition to filamentous cells during biofilm formation by C. albicans. © 2016 Blackwell Verlag GmbH.

  6. The filamentous growth MAPK Pathway Responds to Glucose Starvation Through the Mig1/2 transcriptional repressors in Saccharomyces cerevisiae.

    Science.gov (United States)

    Karunanithi, Sheelarani; Cullen, Paul J

    2012-11-01

    In the budding yeast S. cerevisiae, nutrient limitation induces a MAPK pathway that regulates filamentous growth and biofilm/mat formation. How nutrient levels feed into the regulation of the filamentous growth pathway is not entirely clear. We characterized a newly identified MAPK regulatory protein of the filamentous growth pathway, Opy2. A two-hybrid screen with the cytosolic domain of Opy2 uncovered new interacting partners including a transcriptional repressor that functions in the AMPK pathway, Mig1, and its close functional homolog, Mig2. Mig1 and Mig2 coregulated the filamentous growth pathway in response to glucose limitation, as did the AMP kinase Snf1. In addition to associating with Opy2, Mig1 and Mig2 interacted with other regulators of the filamentous growth pathway including the cytosolic domain of the signaling mucin Msb2, the MAP kinase kinase Ste7, and the MAP kinase Kss1. As for Opy2, Mig1 overproduction dampened the pheromone response pathway, which implicates Mig1 and Opy2 as potential regulators of pathway specificity. Taken together, our findings provide the first regulatory link in yeast between components of the AMPK pathway and a MAPK pathway that controls cellular differentiation.

  7. Structure of the acidianus filamentous virus 3 and comparative genomics of related archaeal lipothrixviruses

    DEFF Research Database (Denmark)

    Vestergaard, Gisle Alberg; Aramayo, Ricardo; Basta, Tamara

    2008-01-01

    Four novel filamentous viruses with double-stranded DNA genomes, namely, Acidianus filamentous virus 3 (AFV3), AFV6, AFV7, and AFV8, have been characterized from the hyperthermophilic archaeal genus Acidianus, and they are assigned to the Betalipothrixvirus genus of the family Lipothrixviridae....... The structures of the approximately 2-mum-long virions are similar, and one of them, AFV3, was studied in detail. It consists of a cylindrical envelope containing globular subunits arranged in a helical formation that is unique for any known double-stranded DNA virus. The envelope is 3.1 nm thick and encases...... structural proteins; (iii) multiple overlapping open reading frames, which may be indicative of gene recoding; (iv) putative 12-bp genetic elements; and (v) partial gene sequences corresponding closely to spacer sequences of chromosomal repeat clusters....

  8. Automated image analysis for quantification of filamentous bacteria

    DEFF Research Database (Denmark)

    Fredborg, M.; Rosenvinge, F. S.; Spillum, E.

    2015-01-01

    Background: Antibiotics of the beta-lactam group are able to alter the shape of the bacterial cell wall, e.g. filamentation or a spheroplast formation. Early determination of antimicrobial susceptibility may be complicated by filamentation of bacteria as this can be falsely interpreted as growth...... displaying different resistant profiles and differences in filamentation kinetics were used to study a novel image analysis algorithm to quantify length of bacteria and bacterial filamentation. A total of 12 beta-lactam antibiotics or beta-lactam-beta-lactamase inhibitor combinations were analyzed...... in systems relying on colorimetry or turbidometry (such as Vitek-2, Phoenix, MicroScan WalkAway). The objective was to examine an automated image analysis algorithm for quantification of filamentous bacteria using the 3D digital microscopy imaging system, oCelloScope. Results: Three E. coli strains...

  9. Bursting of filaments in the plasma focus

    International Nuclear Information System (INIS)

    Gratton, F.T.L.

    1976-01-01

    Photographs of the current sheath of (low energy) plasma focus show a disruption of the filaments. This phenomenon is interpreted as a vortex breakdown. Physical parameters which support this hypothesis are obtained from measurements, from the theoretical thickness of the current sheath given by Nardi and from some models of the plasma flow. The widening of a vortex due to axial velocity increase is analyzed by means of magnetohydrodynamic collinear models. The main results are: (1) the existence of a limit separating supercritical from subcritical regimes (their character changes with the ratio between kinetic and magnetic energy); (2) the existence of flow regimes where the vortex radius remains approximately constant for moderate increments of the external velocity; (3) the structure of the vortex may change substantially for a sufficiently large increment of the external velocity, even in subcritical states; (4) the possibility that a burst of the vortex may occur when the external velocity suffers a slowdown

  10. Methods for genetic transformation of filamentous fungi.

    Science.gov (United States)

    Li, Dandan; Tang, Yu; Lin, Jun; Cai, Weiwen

    2017-10-03

    Filamentous fungi have been of great interest because of their excellent ability as cell factories to manufacture useful products for human beings. The development of genetic transformation techniques is a precondition that enables scientists to target and modify genes efficiently and may reveal the function of target genes. The method to deliver foreign nucleic acid into cells is the sticking point for fungal genome modification. Up to date, there are some general methods of genetic transformation for fungi, including protoplast-mediated transformation, Agrobacterium-mediated transformation, electroporation, biolistic method and shock-wave-mediated transformation. This article reviews basic protocols and principles of these transformation methods, as well as their advantages and disadvantages.

  11. Filament wound data base development, revision 1

    Science.gov (United States)

    Sharp, R. Scott; Braddock, William F.

    1985-01-01

    The objective was to update the present Space Shuttle Solid Rocket Booster (SRB) baseline reentry aerodynamic data base and to develop a new reentry data base for the filament wound case SRB along with individual protuberance increments. Lockheed's procedures for performing these tasks are discussed. Free fall of the SRBs after separation from the Space Shuttle Launch Vehicle is completely uncontrolled. However, the SRBs must decelerate to a velocity and attitude that is suitable for parachute deployment. To determine the SRB reentry trajectory parameters, including the rate of deceleration and attitude history during free-fall, engineers at Marshall Space Flight Center are using a six-degree-of-freedom computer program to predict dynamic behavior. Static stability aerodynamic coefficients are part of the information required for input into this computer program. Lockheed analyzed the existing reentry aerodynamic data tape (Data Tape 5) for the current steel case SRB. This analysis resulted in the development of Data Tape 7.

  12. Validation of the filament winding process model

    Science.gov (United States)

    Calius, Emilo P.; Springer, George S.; Wilson, Brian A.; Hanson, R. Scott

    1987-01-01

    Tests were performed toward validating the WIND model developed previously for simulating the filament winding of composite cylinders. In these tests two 24 in. long, 8 in. diam and 0.285 in. thick cylinders, made of IM-6G fibers and HBRF-55 resin, were wound at + or - 45 deg angle on steel mandrels. The temperatures on the inner and outer surfaces and inside the composite cylinders were recorded during oven cure. The temperatures inside the cylinders were also calculated by the WIND model. The measured and calculated temperatures were then compared. In addition, the degree of cure and resin viscosity distributions inside the cylinders were calculated for the conditions which existed in the tests.

  13. Natural Fiber Filament Wound Composites: A Review

    Directory of Open Access Journals (Sweden)

    Mohamed Ansari Suriyati

    2017-01-01

    Full Text Available In recent development, natural fibers have attracted the interest of engineers, researchers, professionals and scientists all over the world as an alternative reinforcement for fiber reinforced polymer composites. This is due to its superior properties such as high specific strength, low weight, low cost, fairly good mechanical properties, non-abrasive, eco-friendly and bio-degradable characteristics. In this point of view, natural fiber-polymer composites (NFPCs are becoming increasingly utilized in a wide variety of applications because they represent an ecological and inexpensive alternative to conventional petroleum-derived materials. On the other hand, considerable amounts of organic waste and residue from the industrial and agricultural processes are still underutilized as low-value energy sources. This is a comprehensive review discussing about natural fiber reinforced composite produced by filament winding technique.

  14. HIERARCHICAL FRAGMENTATION OF THE ORION MOLECULAR FILAMENTS

    International Nuclear Information System (INIS)

    Takahashi, Satoko; Ho, Paul T. P.; Su, Yu-Nung; Teixeira, Paula S.; Zapata, Luis A.

    2013-01-01

    We present a high angular resolution map of the 850 μm continuum emission of the Orion Molecular Cloud-3 (OMC 3) obtained with the Submillimeter Array (SMA); the map is a mosaic of 85 pointings covering an approximate area of 6.'5 × 2.'0 (0.88 × 0.27 pc). We detect 12 spatially resolved continuum sources, each with an H 2 mass between 0.3-5.7 M ☉ and a projected source size between 1400-8200 AU. All the detected sources are on the filamentary main ridge (n H 2 ≥10 6 cm –3 ), and analysis based on the Jeans theorem suggests that they are most likely gravitationally unstable. Comparison of multi-wavelength data sets indicates that of the continuum sources, 6/12 (50%) are associated with molecular outflows, 8/12 (67%) are associated with infrared sources, and 3/12 (25%) are associated with ionized jets. The evolutionary status of these sources ranges from prestellar cores to protostar phase, confirming that OMC-3 is an active region with ongoing embedded star formation. We detect quasi-periodical separations between the OMC-3 sources of ≈17''/0.035 pc. This spatial distribution is part of a large hierarchical structure that also includes fragmentation scales of giant molecular cloud (≈35 pc), large-scale clumps (≈1.3 pc), and small-scale clumps (≈0.3 pc), suggesting that hierarchical fragmentation operates within the Orion A molecular cloud. The fragmentation spacings are roughly consistent with the thermal fragmentation length in large-scale clumps, while for small-scale cores it is smaller than the local fragmentation length. These smaller spacings observed with the SMA can be explained by either a helical magnetic field, cloud rotation, or/and global filament collapse. Finally, possible evidence for sequential fragmentation is suggested in the northern part of the OMC-3 filament.

  15. RADIATION SPECTRAL SYNTHESIS OF RELATIVISTIC FILAMENTATION

    International Nuclear Information System (INIS)

    Frederiksen, Jacob Trier; Haugboelle, Troels; Medvedev, Mikhail V.; Nordlund, Ake

    2010-01-01

    Radiation from many astrophysical sources, e.g., gamma-ray bursts and active galactic nuclei, is believed to arise from relativistically shocked collisionless plasmas. Such sources often exhibit highly transient spectra evolving rapidly compared with source lifetimes. Radiation emitted from these sources is typically associated with nonlinear plasma physics, complex field topologies, and non-thermal particle distributions. In such circumstances, a standard synchrotron paradigm may fail to produce accurate conclusions regarding the underlying physics. Simulating spectral emission and spectral evolution numerically in various relativistic shock scenarios is then the only viable method to determine the detailed physical origin of the emitted spectra. In this Letter, we present synthetic radiation spectra representing the early stage development of the filamentation (streaming) instability of an initially unmagnetized plasma, which is relevant for both collisionless shock formation and reconnection dynamics in relativistic astrophysical outflows as well as for laboratory astrophysics experiments. Results were obtained using a highly efficient in situ diagnostics method, based on detailed particle-in-cell modeling of collisionless plasmas. The synthetic spectra obtained here are compared with those predicted by a semi-analytical model for jitter radiation from the filamentation instability, the latter including self-consistent generated field topologies and particle distributions obtained from the simulations reported upon here. Spectra exhibit dependence on the presence-or the absence-of an inert plasma constituent, when comparing baryonic plasmas (i.e., containing protons) with pair plasmas. The results also illustrate that considerable care should be taken when using lower-dimensional models to obtain information about the astrophysical phenomena generating observed spectra.

  16. Bundling of elastic filaments induced by hydrodynamic interactions

    Science.gov (United States)

    Man, Yi; Page, William; Poole, Robert J.; Lauga, Eric

    2017-12-01

    Peritrichous bacteria swim in viscous fluids by rotating multiple helical flagellar filaments. As the bacterium swims forward, all its flagella rotate in synchrony behind the cell in a tight helical bundle. When the bacterium changes its direction, the flagellar filaments unbundle and randomly reorient the cell for a short period of time before returning to their bundled state and resuming swimming. This rapid bundling and unbundling is, at its heart, a mechanical process whereby hydrodynamic interactions balance with elasticity to determine the time-varying deformation of the filaments. Inspired by this biophysical problem, we present in this paper what is perhaps the simplest model of bundling whereby two or more straight elastic filaments immersed in a viscous fluid rotate about their centerline, inducing rotational flows which tend to bend the filaments around each other. We derive an integrodifferential equation governing the shape of the filaments resulting from mechanical balance in a viscous fluid at low Reynolds number. We show that such equation may be evaluated asymptotically analytically in the long-wavelength limit, leading to a local partial differential equation governed by a single dimensionless bundling number. A numerical study of the dynamics predicted by the model reveals the presence of two configuration instabilities with increasing bundling numbers: first to a crossing state where filaments touch at one point and then to a bundled state where filaments wrap along each other in a helical fashion. We also consider the case of multiple filaments and the unbundling dynamics. We next provide an intuitive physical model for the crossing instability and show that it may be used to predict analytically its threshold and adapted to address the transition to a bundling state. We then use a macroscale experimental implementation of the two-filament configuration in order to validate our theoretical predictions and obtain excellent agreement. This long

  17. Comparative Genome Analysis of Filamentous Fungi Reveals Gene Family Expansions Associated with Fungal Pathogenesis

    Science.gov (United States)

    Soanes, Darren M.; Alam, Intikhab; Cornell, Mike; Wong, Han Min; Hedeler, Cornelia; Paton, Norman W.; Rattray, Magnus; Hubbard, Simon J.; Oliver, Stephen G.; Talbot, Nicholas J.

    2008-01-01

    Fungi and oomycetes are the causal agents of many of the most serious diseases of plants. Here we report a detailed comparative analysis of the genome sequences of thirty-six species of fungi and oomycetes, including seven plant pathogenic species, that aims to explore the common genetic features associated with plant disease-causing species. The predicted translational products of each genome have been clustered into groups of potential orthologues using Markov Chain Clustering and the data integrated into the e-Fungi object-oriented data warehouse (http://www.e-fungi.org.uk/). Analysis of the species distribution of members of these clusters has identified proteins that are specific to filamentous fungal species and a group of proteins found only in plant pathogens. By comparing the gene inventories of filamentous, ascomycetous phytopathogenic and free-living species of fungi, we have identified a set of gene families that appear to have expanded during the evolution of phytopathogens and may therefore serve important roles in plant disease. We have also characterised the predicted set of secreted proteins encoded by each genome and identified a set of protein families which are significantly over-represented in the secretomes of plant pathogenic fungi, including putative effector proteins that might perturb host cell biology during plant infection. The results demonstrate the potential of comparative genome analysis for exploring the evolution of eukaryotic microbial pathogenesis. PMID:18523684

  18. Chaperone heat shock protein 90 mobilization and hydralazine cytoprotection against acrolein-induced carbonyl stress.

    Science.gov (United States)

    Burcham, Philip C; Raso, Albert; Kaminskas, Lisa M

    2012-11-01

    Toxic carbonyls such as acrolein participate in many degenerative diseases. Although the nucleophilic vasodilatory drug hydralazine readily traps such species under "test-tube" conditions, whether these reactions adequately explain its efficacy in animal models of carbonyl-mediated disease is uncertain. We have previously shown that hydralazine attacks carbonyl-adducted proteins in an "adduct-trapping" reaction that appears to take precedence over direct "carbonyl-sequestering" reactions, but how this reaction conferred cytoprotection was unclear. This study explored the possibility that by increasing the bulkiness of acrolein-adducted proteins, adduct-trapping might alter the redistribution of chaperones to damaged cytoskeletal proteins that are known targets for acrolein. Using A549 lung adenocarcinoma cells, the levels of chaperones heat shock protein (Hsp) 40, Hsp70, Hsp90, and Hsp110 were measured in intermediate filament extracts prepared after a 3-h exposure to acrolein. Exposure to acrolein alone modestly increased the levels of all four chaperones. Coexposure to hydralazine (10-100 μM) strongly suppressed cell ATP loss while producing strong adduct-trapping in intermediate filaments. Most strikingly, hydralazine selectively boosted the levels of cytoskeletal-associated Hsp90, including a high-mass species that was sensitive to the Hsp90 inhibitor 17-N-allylamino-17-demethoxygeldanamycin. Biochemical fractionation of acrolein- and hydralazine-treated cells revealed that hydralazine likely promoted Hsp90 migration from cytosol into other subcellular compartments. A role for Hsp90 mobilization in cytoprotection was confirmed by the finding that brief heat shock treatment suppressed acute acrolein toxicity in A549 cells. Taken together, these findings suggest that by increasing the steric bulk of carbonyl-adducted proteins, adduct-trapping drugs trigger the intracellular mobilization of the key molecular chaperone Hsp90.

  19. O-sulfonation of serine and threonine: mass spectrometric detection and characterization of a new posttranslational modification in diverse proteins throughout the eukaryotes.

    Science.gov (United States)

    Medzihradszky, K F; Darula, Z; Perlson, E; Fainzilber, M; Chalkley, R J; Ball, H; Greenbaum, D; Bogyo, M; Tyson, D R; Bradshaw, R A; Burlingame, A L

    2004-05-01

    Protein sulfonation on serine and threonine residues is described for the first time. This post-translational modification is shown to occur in proteins isolated from organisms representing a broad span of eukaryote evolution, including the invertebrate mollusk Lymnaea stagnalis, the unicellular malaria parasite Plasmodium falciparum, and humans. Detection and structural characterization of this novel post-translational modification was carried out using liquid chromatography coupled to electrospray tandem mass spectrometry on proteins including a neuronal intermediate filament and a myosin light chain from the snail, a cathepsin-C-like enzyme from the parasite, and the cytoplasmic domain of the human orphan receptor tyrosine kinase Ror-2. These findings suggest that sulfonation of serine and threonine may be involved in multiple functions including protein assembly and signal transduction.

  20. EST Table: FS851943 [KAIKOcDNA[Archive

    Lifescience Database Archive (English)

    Full Text Available ar to restin (Reed-Steinberg cell-expressed intermediate filament-associated protein) [Tribolium castaneum] ...o restin (Reed-Steinberg cell-expressed intermediate filament-associated protein) [Tribolium castaneum] FS906662 fner ...

  1. EST Table: FS921686 [KAIKOcDNA[Archive

    Lifescience Database Archive (English)

    Full Text Available ar to restin (Reed-Steinberg cell-expressed intermediate filament-associated protein) [Tribolium castaneum] ...o restin (Reed-Steinberg cell-expressed intermediate filament-associated protein) [Tribolium castaneum] FS906662 fwgP ...

  2. Top-down label-free LC-MALDI analysis of the peptidome during neural progenitor cell differentiation reveals complexity in cytoskeletal protein dynamics and identifies progenitor cell markers.

    Science.gov (United States)

    Maltman, Daniel J; Brand, Sven; Belau, Eckhard; Paape, Rainer; Suckau, Detlev; Przyborski, Stefan A

    2011-10-01

    In the field of stem cell research, there is a strong requirement for the discovery of new biomarkers that more accurately define stem and progenitor cell populations, as well as their differentiated derivatives. The very-low-molecular-weight (differential abundance between undifferentiated and differentiated cultures. These proteins included major cytoskeletal components such as nestin, vimentin, and glial fibrillary acidic protein, which are all associated with neural development. Other cytoskeletal proteins identified were dihydropyrimidinase-related protein 2, prothymosin (thymosin α-1), and thymosin β-10. These findings highlight novel stem cell/progenitor cell marker candidates and demonstrate proteomic complexity, which underlies the limitations of major intermediate filament proteins long established as neural markers. Copyright © 2011 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  3. Noncovalent Intermediate of Thymidylate Synthase: Fact or Fiction?

    Science.gov (United States)

    Kholodar, Svetlana A; Kohen, Amnon

    2016-07-06

    Thymidylate synthase is an attractive target for antibiotic and anticancer drugs due to its essential role in the de novo biosynthesis of the DNA nucleotide thymine. The enzymatic reaction is initiated by a nucleophilic activation of the substrate via formation of a covalent bond to an active site cysteine. The traditionally accepted mechanism is then followed by a series of covalently bound intermediates, where that bond is only cleaved upon product release. Recent computational and experimental studies suggest that the covalent bond between the protein and substrate is actually quite labile. Importantly, these findings predict the existence of a noncovalently bound bisubstrate intermediate, not previously anticipated, which could be the target of a novel class of drugs inhibiting DNA biosynthesis. Here we report the synthesis of the proposed intermediate and findings supporting its chemical and kinetic competence. These findings substantiate the predicted nontraditional mechanism and the potential of this intermediate as a new drug lead.

  4. Assembling filamentous phage occlude pIV channels.

    Science.gov (United States)

    Marciano, D K; Russel, M; Simon, S M

    2001-07-31

    Filamentous phage f1 is exported from its Escherichia coli host without killing the bacterial cell. Phage-encoded protein pIV, which is required for phage assembly and secretion, forms large highly conductive channels in the outer membrane of E. coli. It has been proposed that the phage are extruded across the bacterial outer membrane through pIV channels. To test this prediction, we developed an in vivo assay by using a mutant pIV that functions in phage export but whose channel opens in the absence of phage extrusion. In E. coli lacking its native maltooligosacharride transporter LamB, this pIV variant allowed oligosaccharide transport across the outer membrane. This entry of oligosaccharide was decreased by phage production and still further decreased by production of phage that cannot be released from the cell surface. Thus, exiting phage block the pIV-dependent entry of oligosaccharide, suggesting that phage occupy the lumen of pIV channels. This study provides the first evidence, to our knowledge, for viral exit through a large aqueous channel.

  5. Mechanisms of deterioration of intermediate moisture food systems

    Science.gov (United States)

    Labuza, T. P.

    1972-01-01

    A study of shelf stability in intermediate moisture foods was made. Major efforts were made to control lipid oxidation and nonenzymatic browning. In order to determine means of preventing these reactions, model systems were developed having the same water activity content relationship of intermediate moisture foods. Models were based on a cellulose-lipid and protein-lipid system with glycerol added as the humectant. Experiments with both systems indicate that lipid oxidation is promoted significantly in the intermediate moisture range. The effect appeared to be related to increased mobility of either reactants or catalysts, since when the amount of water in the system reached a level where capillary condensation occurred and thus free water was present, the rates of oxidation increased. With added glycerol, which is water soluble and thus increases the amount of mobile phase, the increase in oxidation rate occurs at a lower relative humidity. The rates of oxidation were maximized at 61% RH and decreased again at 75% RH probably due to dilution. No significant non-enzymatic browning occurred in the protein-lipid systems. Prevention of oxidation by the use of metal chelating agents was enhanced in the cellulose system, whereas, with protein present, the lipid soluble chain terminating antioxidants (such as BHA) worked equally as well. Preliminary studies of foods adjusted to the intermediate moisture range bear out the results of oxidation in model systems. It can be concluded that for most fat containing intermediate moisture foods, rancidity will be the reaction most limiting stability.

  6. Harmful impact of filamentous algae (Spirogyra sp. on juvenile crayfish

    Directory of Open Access Journals (Sweden)

    Ulikowski Dariusz

    2015-12-01

    Full Text Available The aim of this study was to determine the impact of filamentous algae on the growth and survival of juvenile narrow-clawed crayfish, Astacus leptodactylus (Esch., in rearing basins. Three stocking variants were used: A - basins with a layer of filamentous algae without imitation mineral substrate; B - basins with a layer of filamentous algae with imitation mineral substrate; C - basins without filamentous algae but with mineral substrate. The crayfish were reared from June 12 to October 10 under natural thermal conditions and fed a commercial feed. The results indicated that the presence of the filamentous algae did not have a statistically significant impact on the growth of the juvenile crayfish (P > 0.05. The presence of the filamentous algae had a strong negative impact on juvenile crayfish survival and stock biomass (P < 0.05. The layer of gravel and small stones that imitated the mineral substrate of natural aquatic basins somewhat neutralized the disadvantageous impact the filamentous algae had on the crayfish.

  7. Footpoint detection and mass-motion in chromospheric filaments

    Science.gov (United States)

    V, Aparna; Hardersen, P. S.; Martin, S. F.

    2013-07-01

    A quiescent region on the Sun containing three filaments is used to study the properties of mass motion. This study determines if the footpoints or end-points of the filaments are the locations from where mass gets injected into the filaments. Several hypotheses have been put forth in the past to determine how a filament acquires mass. Trapping of coronal mass in the filament channel due to condensation (Martin, 1996) and injection of mass into the filaments during magnetic reconnection (Priest, et al., 1995) are some of the speculations. This study looks for indications for injection of mass via chromospheric footpoints. The data consists of blue (Hα-0.5 Å) and red (Hα+0.5 Å) wing high resolution Hα images of the W29N37 region of the Sun taken on Oct 30, 2010, from 1200 - 1600 UT. The Dutch Open Telescope was used to obtain the data. The images are aligned and animated to see Doppler motion in the fibrils. Smaller fibrils merge to form longer ones; barbs appear and disappear in one of the long filaments and is seen moving along the length of the filament. A region with no typical filament-like absorption feature is observed to be continuously receiving mass. Fibrils appear to be converging from opposite sides along what appears to be a neutral line; mass motion is seen in these fibrils as well. An eruption occurs in a region of fibrils lumped together at the end of the first hour (1300 UT) followed by plage brightening at 1430 UT near one of the filament regions. Helioviewer (Panasenco, et al., 2011) is used for aligning the images; GIMP is used for precision alignment and animation. Each frame in the sequence is studied carefully to note changes in the filament regions. The footpoints of the filaments are determined by the changes observed in the position of the filament ‘legs’ in each frame. Variations in the magnetic polarity corresponding to changes observed in the chromosphere are analyzed using HMI magnetograms. Bright and dark points on the

  8. Microwave structure of quiescent solar filaments at high resolution

    International Nuclear Information System (INIS)

    Gary, D.E.

    1986-01-01

    High resolution very low altitude maps of a quiescent filament at three frequencies are presented. The spatial resolution (approx. 15'' at 1.45 GHz, approx. 6'' at 4.9 GHz, and approx. 2'' at 15 GHz) is several times better than previously attained. At each frequency, the filament appears as a depression in the quiet Sun background. The depression is measurably wider and longer in extent than the corresponding H alpha filament at 1.45 GHz and 4.9 GHz, indicating that the depression is due in large part to a deficit in coronal density associated with the filament channel. In contrast, the shape of the radio depression at 15 CHz closely matches that of the H alpha filament. In addition, the 15 GHz map shows enhanced emission along both sides of the radio depression. A similar enhancement is seen in an observation of a second filament 4 days later, which suggests that the enhancement is a general feature of filaments. Possible causes of the enhanced emission are explored

  9. A study of short wave instability on vortex filaments

    Energy Technology Data Exchange (ETDEWEB)

    Wang, Hong Yun [Univ. of California, Berkeley, CA (United States)

    1996-12-01

    The numerical stability and accuracy of the vortex method are studied. The effect of the ordinary differential equations (ODE) solver and of the time step on the numerical stability is analyzed. Various ODE solvers are compared and a best performer is chosen. A new constraint on the time step based on numerical stability is proposed and verified in numerical simulations. It is shown through numerical examples that empirical rules for selecting the spatial discretization obtained in simple test problems may not be extended to more general problems. The thin tube vortex filament method is applied to the problem of Widnall's instability on vortex rings. Numerical results different from previous calculations are presented and the source of the discrepancies is explained. The long time behavior of the unstable mode on thin vortex rings is simulated and analyzed. The short wave instability on vortex filaments is investigated both theoretically and numerically. It is shown that the short wave instability always occurs on co-rotating vortex filaments of fixed core structure. Furthermore when they are close to each other, vortex filaments produce short wave unstable modes which lead to wild stretching and folding. However, when the inter-filament distance is large in comparison with the core size of the filaments, unstable modes are bounded by a small fraction of the core size and the vortex filaments do not create hairpins nor wild stretching. These findings may explain the smooth behavior of the superfluid vortices. The formation of hairpin structures on numerical vortex filaments is investigated. It is shown that the formation of hairpin structures is independent of the ODE solver, of the time step and of other numerical parameters. The hairpin structures are primarily caused by short wave instability on co-rotating vortex filaments.

  10. Gravity with Intermediate Goods Trade

    Directory of Open Access Journals (Sweden)

    Sujin Jang

    2017-12-01

    Full Text Available This paper derives the gravity equation with intermediate goods trade. We extend a standard monopolistic competition model to incorporate intermediate goods trade, and show that the gravity equation with intermediates trade is identical to the one without it except in that gross output should be used as the output measure instead of value added. We also show that the output elasticity of trade is significantly underestimated when value added is used as the output measure. This implies that with the conventional gravity equation, the contribution of output growth can be substantially underestimated and the role of trade costs reduction can be exaggerated in explaining trade expansion, as we demonstrate for the case of Korea's trade growth between 1995 and 2007.

  11. Larval helminths in intermediate hosts

    DEFF Research Database (Denmark)

    Fredensborg, Brian Lund; Poulin, R

    2005-01-01

    Density-dependent effects on parasite fitness have been documented from adult helminths in their definitive hosts. There have, however, been no studies on the cost of sharing an intermediate host with other parasites in terms of reduced adult parasite fecundity. Even if larval parasites suffer...... transmission to their bird definitive host by predation. In experimental infections, we found an intensity-dependent establishment success, with a decrease in the success rate of cercariae developing into infective metacercariae with an increasing dose of cercariae applied to each amphipod. In natural...... the two species. Our results thus indicate that the infracommunity of larval helminths in their intermediate host is interactive and that any density-dependent effect in the intermediate host may have lasting effects on individual parasite fitness....

  12. HIERARCHICAL FRAGMENTATION OF THE ORION MOLECULAR FILAMENTS

    Energy Technology Data Exchange (ETDEWEB)

    Takahashi, Satoko; Ho, Paul T. P.; Su, Yu-Nung [Academia Sinica Institute of Astronomy and Astrophysics, P.O. Box 23-141, Taipei 10617, Taiwan (China); Teixeira, Paula S. [Institut fuer Astrophysik, Universitaet Wien, Tuerkenschanzstrasse 17, A-1180, Wien (Austria); Zapata, Luis A., E-mail: satoko_t@asiaa.sinica.edu.tw [Centro de Radioastronomia y Astrofisica, Universidad Nacional Autonoma de Mexico, Morelia, Michoacan 58090 (Mexico)

    2013-01-20

    We present a high angular resolution map of the 850 {mu}m continuum emission of the Orion Molecular Cloud-3 (OMC 3) obtained with the Submillimeter Array (SMA); the map is a mosaic of 85 pointings covering an approximate area of 6.'5 Multiplication-Sign 2.'0 (0.88 Multiplication-Sign 0.27 pc). We detect 12 spatially resolved continuum sources, each with an H{sub 2} mass between 0.3-5.7 M {sub Sun} and a projected source size between 1400-8200 AU. All the detected sources are on the filamentary main ridge (n{sub H{sub 2}}{>=}10{sup 6} cm{sup -3}), and analysis based on the Jeans theorem suggests that they are most likely gravitationally unstable. Comparison of multi-wavelength data sets indicates that of the continuum sources, 6/12 (50%) are associated with molecular outflows, 8/12 (67%) are associated with infrared sources, and 3/12 (25%) are associated with ionized jets. The evolutionary status of these sources ranges from prestellar cores to protostar phase, confirming that OMC-3 is an active region with ongoing embedded star formation. We detect quasi-periodical separations between the OMC-3 sources of Almost-Equal-To 17''/0.035 pc. This spatial distribution is part of a large hierarchical structure that also includes fragmentation scales of giant molecular cloud ( Almost-Equal-To 35 pc), large-scale clumps ( Almost-Equal-To 1.3 pc), and small-scale clumps ( Almost-Equal-To 0.3 pc), suggesting that hierarchical fragmentation operates within the Orion A molecular cloud. The fragmentation spacings are roughly consistent with the thermal fragmentation length in large-scale clumps, while for small-scale cores it is smaller than the local fragmentation length. These smaller spacings observed with the SMA can be explained by either a helical magnetic field, cloud rotation, or/and global filament collapse. Finally, possible evidence for sequential fragmentation is suggested in the northern part of the OMC-3 filament.

  13. Failure and nonfailure of fluid filaments in extension

    DEFF Research Database (Denmark)

    Hassager, Ole; Kolte, Mette Irene; Renardy, Michael

    1998-01-01

    The phenomenon of ductile failure of Newtonian and viscoelastic fluid filaments without surface tension is studied by a 2D finite element method and by ID non-linear analysis. The viscoelastic fluids are described by single integral constitutive equations. The main conclusions are: (1) Newtonian...... fluid filaments do not exhibit ductile failure without surface tension; (2) some viscoelastic fluids form stable filaments while other fluids exhibit ductile failure as a result of an elastic instability; (3) for large Deborah numbers, the Considere condition may be used to predict the Hencky strain...

  14. Dynamics of actin cables in polarized growth of the filamentous fungus Aspergillus nidulans

    Directory of Open Access Journals (Sweden)

    Anna eBergs

    2016-05-01

    Full Text Available Highly polarized growth of filamentous fungi requires a continuous supply of proteins and lipids to the hyphal tip. This transport is managed by vesicle trafficking via the actin and microtubule cytoskeletons and their associated motor proteins. Particularly, actin cables originating from the hyphal tip are essential for hyphal growth. Although specific marker proteins to visualize actin cables have been developed in filamentous fungi, the exact organization and dynamics of actin cables has remained elusive. Here we visualized actin cables using tropomyosin (TpmA and Lifeact fused to fluorescent proteins in Aspergillus nidulans and studied the dynamics and regulation. GFP tagged TpmA visualized dynamic actin cables formed from the hyphal tip with cycles of elongation and shrinkage. The elongation and shrinkage rates of actin cables were similar and approximately 0.6 μm/s. Comparison of actin markers revealed that high concentrations of Lifeact reduced actin dynamics. Simultaneous visualization of actin cables and microtubules suggests temporally and spatially coordinated polymerization and depolymerization between the two cytoskeletons. Our results provide new insights into the molecular mechanism of ordered polarized growth regulated by actin cables and microtubules.

  15. A core filamentation response network in Candida albicans is restricted to eight genes.

    Directory of Open Access Journals (Sweden)

    Ronny Martin

    Full Text Available Although morphological plasticity is a central virulence trait of Candida albicans, the number of filament-associated genes and the interplay of mechanisms regulating their expression remain unknown. By correlation-based network modeling of the transcriptional response to different defined external stimuli for morphogenesis we identified a set of eight genes with highly correlated expression patterns, forming a core filamentation response. This group of genes included ALS3, ECE1, HGT2, HWP1, IHD1 and RBT1 which are known or supposed to encode for cell- wall associated proteins as well as the Rac1 guanine nucleotide exchange factor encoding gene DCK1 and the unknown function open reading frame orf19.2457. The validity of network modeling was confirmed using a dataset of advanced complexity that describes the transcriptional response of C. albicans during epithelial invasion as well as comparing our results with other previously published transcriptome studies. Although the set of core filamentation response genes was quite small, several transcriptional regulators are involved in the control of their expression, depending on the environmental condition.

  16. A mathematical model of actin filament turnover for fitting FRAP data.

    Science.gov (United States)

    Halavatyi, Aliaksandr A; Nazarov, Petr V; Al Tanoury, Ziad; Apanasovich, Vladimir V; Yatskou, Mikalai; Friederich, Evelyne

    2010-03-01

    A novel mathematical model of the actin dynamics in living cells under steady-state conditions has been developed for fluorescence recovery after photobleaching (FRAP) experiments. As opposed to other FRAP fitting models, which use the average lifetime of actins in filaments and the actin turnover rate as fitting parameters, our model operates with unbiased actin association/dissociation rate constants and accounts for the filament length. The mathematical formalism is based on a system of stochastic differential equations. The derived equations were validated on synthetic theoretical data generated by a stochastic simulation algorithm adapted for the simulation of FRAP experiments. Consistent with experimental findings, the results of this work showed that (1) fluorescence recovery is a function of the average filament length, (2) the F-actin turnover and the FRAP are accelerated in the presence of actin nucleating proteins, (3) the FRAP curves may exhibit both a linear and non-linear behaviour depending on the parameters of actin polymerisation, and (4) our model resulted in more accurate parameter estimations of actin dynamics as compared with other FRAP fitting models. Additionally, we provide a computational tool that integrates the model and that can be used for interpretation of FRAP data on actin cytoskeleton.

  17. The Structure of HIV-1 Rev Filaments Suggests a Bilateral Model for Rev-RRE Assembly.

    Science.gov (United States)

    DiMattia, Michael A; Watts, Norman R; Cheng, Naiqian; Huang, Rick; Heymann, J Bernard; Grimes, Jonathan M; Wingfield, Paul T; Stuart, David I; Steven, Alasdair C

    2016-07-06

    HIV-1 Rev protein mediates the nuclear export of viral RNA genomes. To do so, Rev oligomerizes cooperatively onto an RNA motif, the Rev response element (RRE), forming a complex that engages with the host nuclear export machinery. To better understand Rev oligomerization, we determined four crystal structures of Rev N-terminal domain dimers, which show that they can pivot about their dyad axis, giving crossing angles of 90° to 140°. In parallel, we performed cryoelectron microscopy of helical Rev filaments. Filaments vary from 11 to 15 nm in width, reflecting variations in dimer crossing angle. These structures contain additional density, indicating that C-terminal domains become partially ordered in the context of filaments. This conformational variability may be exploited in the assembly of RRE/Rev complexes. Our data also revealed a third interface between Revs, which offers an explanation for how the arrangement of Rev subunits adapts to the "A"-shaped architecture of the RRE in export-active complexes. Published by Elsevier Ltd.

  18. Viscoelastic behavior of human lamin A proteins in the context of dilated cardiomyopathy.

    Directory of Open Access Journals (Sweden)

    Avinanda Banerjee

    Full Text Available Lamins are intermediate filament proteins of type V constituting a nuclear lamina or filamentous meshwork which lines the nucleoplasmic side of the inner nuclear membrane. This protein mesh provides a supporting scaffold for the nuclear envelope and tethers interphase chromosome to the nuclear periphery. Mutations of mainly A-type lamins are found to be causative for at least 11 human diseases collectively termed as laminopathies majority of which are characterised by aberrant nuclei with altered structural rigidity, deformability and poor mechanotransduction behaviour. But the investigation of viscoelastic behavior of lamin A continues to elude the field. In order to address this problem, we hereby present the very first report on viscoelastic properties of wild type human lamin A and some of its mutants linked with Dilated cardiomyopathy (DCM using quantitative rheological measurements. We observed a dramatic strain-softening effect on lamin A network as an outcome of the strain amplitude sweep measurements which could arise from the large compliance of the quasi-cross-links in the network or that of the lamin A rods. In addition, the drastic stiffening of the differential elastic moduli on superposition of rotational and oscillatory shear stress reflect the increase in the stiffness of the laterally associated lamin A rods. These findings present a preliminary insight into distinct biomechanical properties of wild type lamin A protein and its mutants which in turn revealed interesting differences.

  19. Biochemical responses of filamentous algae in different aquatic ecosystems in South East Turkey and associated water quality parameters.

    Science.gov (United States)

    Çelekli, Abuzer; Arslanargun, Hamdullah; Soysal, Çiğdem; Gültekin, Emine; Bozkurt, Hüseyin

    2016-11-01

    To the best of our knowledge, any study about biochemical response of filamentous algae in the complex freshwater ecosystems has not been found in the literature. This study was designed to explore biochemical response of filamentous algae in different water bodies from May 2013 to October 2014, using multivariate approach in the South East of Turkey. Environmental variables were measured in situ: water temperature, oxygen concentration, saturation, conductivity, salinity, pH, redox potential, and total dissolved solid. Chemical variables of aqueous samples and biochemical compounds of filamentous algae were also measured. It was found that geographic position and anthropogenic activities had strong effect on physico-chemical variables of water bodies. Variation in environmental conditions caused change in algal biomass composition due to the different response of filamentous species, also indicated by FTIR analysis. Biochemical responses not only changed from species to species, but also varied for the same species at different sampling time and sampling stations. Multivariate analyses showed that heavy metals, nutrients, and water hardness were found as the important variables governing the temporal and spatial succession and biochemical compounds. Nutrients, especially nitrate, could stimulate pigment and total protein production, whereas high metal content had adverse effects. Amount of malondialdehyde (MDA), H2O2, total thiol groups, total phenolic compounds, proline, total carbohydrate, and metal bioaccumulation by filamentous algae could be closely related with heavy metals in the ecosystems. Significant increase in MDA, H2O2, total thiol group, total phenolic compounds, and proline productions by filamentous algae and chlorosis phenomenon seemed to be an important strategy for alleviating environmental factors-induced oxidative stress as biomarkers. Copyright © 2016 Elsevier Inc. All rights reserved.

  20. Linear viscoelastic characterization from filament stretching rheometry

    DEFF Research Database (Denmark)

    Wingstrand, Sara Lindeblad; Alvarez, Nicolas J.; Hassager, Ole

    Traditionally, linear viscoelasticity is measured using small amplitude oscillatory shear flow. Due to experimental difficulties, shear flows are predominately confined to the linear and mildly nonlinear regime. On the other hand, extensional flows have proven more practical in measuring viscoela......Traditionally, linear viscoelasticity is measured using small amplitude oscillatory shear flow. Due to experimental difficulties, shear flows are predominately confined to the linear and mildly nonlinear regime. On the other hand, extensional flows have proven more practical in measuring...... viscoelasticity well into the nonlinear regime. Therefore at present, complete rheological characterization of a material requires two apparatuses: a shear and an extensional rheometer. This work is focused on developing a linear viscoelastic protocol for the filament stretching rheometer (FSR) in order...... to measure both linear and nonlinear dynamics on a single apparatus. With a software modification to the FSR motor control, we show that linear viscoelasticity can be measured via small amplitude squeeze flow (SASF). Squeeze flow is a combination of both shear and extensional flow applied by axially...

  1. One ring (or two) to hold them all – on the structure and function of protein nanotubes.

    Science.gov (United States)

    Angelucci, Francesco; Bellelli, Andrea; Ardini, Matteo; Ippoliti, Rodolfo; Saccoccia, Fulvio; Morea, Veronica

    2015-08-01

    Understanding the structural determinants relevant to the formation of supramolecular assemblies of homo-oligomeric proteins is a traditional and central scope of structural biology. The knowledge thus gained is crucial both to infer their physiological function and to exploit their architecture for bionanomaterials design. Protein nanotubes made by one-dimensional arrays of homo-oligomers can be generated by either a commutative mechanism, yielding an 'open' structure (e.g. actin), or a noncommutative mechanism, whereby the final structure is formed by hierarchical self-assembly of intermediate 'closed' structures. Examples of the latter process are poorly described and the rules by which they assemble have not been unequivocally defined. We have collected and investigated examples of homo-oligomeric circular arrangements that form one-dimensional filaments of stacked rings by the noncommutative mechanism in vivo and in vitro. Based on their quaternary structure, circular arrangements of protein subunits can be subdivided into two groups that we term Rings of Dimers (e.g. peroxiredoxin and stable protein 1) and Dimers of Rings (e.g. thermosome/rosettasome), depending on the sub-structures that can be identified within the assembly (and, in some cases, populated in solution under selected experimental conditions). Structural analysis allowed us to identify the determinants by which ring-like molecular chaperones form filamentous-like assemblies and to formulate a novel hypothesis by which nanotube assembly, molecular chaperone activity and macromolecular crowding may be interconnected. © 2015 FEBS.

  2. Abstracts of Workshop on Reactive Intermediates in Sulfur Chemistry

    International Nuclear Information System (INIS)

    1998-01-01

    There is a review of the studies on sulfur containing inorganic and organic compounds with special emphasis on biologically important compounds as amino acids and proteins. Photochemically and radiation induced reactive intermediates such as solvated electrons, ions, radicals and radical-ions have been broadly investigated. Their reaction mechanisms and kinetics have been studied and discussed as well

  3. Yield and organoleptic characteristics of Suya (an intermediate ...

    African Journals Online (AJOL)

    High cost of choice meat used in suya production makes the product an exclusive meat for the rich. In order to make suya (an intermediate moisture meat) available and affordable to the common man thereby increasing their animal protein intake, this experiment therefore become imperative. Meat from the ...

  4. Intermediate Infrastructure Analyst | IDRC - International ...

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

    The incumbent conducts research on technologies and tools that might enhance service delivery and where appropriate, makes recommendations to management. The Intermediate Infrastructure System Analyst provides leadership and direction to junior team members and functional direction to consultants and ...

  5. Screening of epoxy systems for high performance filament winding applications

    Science.gov (United States)

    Chiao, T. T.; Jessop, E. S.; Penn, L.

    1975-01-01

    Several promising epoxy systems for high performance filament winding applications are described. Viscosities, gel times, and cast resin tensile behavior are given, as well as heat deflection under load and water absorption measurements.

  6. Biofilms from a Brazilian water distribution system include filamentous fungi.

    Science.gov (United States)

    Siqueira, V M; Oliveira, H M B; Santos, C; Paterson, R R M; Gusmão, N B; Lima, N

    2013-03-01

    Filamentous fungi in drinking water can block water pipes, can cause organoleptic biodeterioration, and are a source of pathogens. There are increasing reports of the involvement of the organisms in biofilms. This present study describes a sampling device that can be inserted directly into pipes within water distribution systems, allowing biofilm formation in situ. Calcofluor White M2R staining and fluorescent in situ hybridization with morphological analyses using epifluorescent microscopy were used to analyse biofilms for filamentous fungi, permitting direct observation of the fungi. DAPI (4',6-diamidino-2-phenylindole) was applied to detect bacteria. Filamentous fungi were detected in biofilms after 6 months on coupons exposed to raw water, decanted water and at the entrance of the water distribution system. Algae, yeast, and bacteria were also observed. The role of filamentous fungi requires further investigations.

  7. Health Risks Associated with Exposure to Filamentous Fungi

    Science.gov (United States)

    Egbuta, Mary Augustina; Mwanza, Mulunda

    2017-01-01

    Filamentous fungi occur widely in the environment, contaminating soil, air, food and other substrates. Due to their wide distribution, they have medical and economic implications. Regardless of their use as a source of antibiotics, vitamins and raw materials for various industrially important chemicals, most fungi and filamentous fungi produce metabolites associated with a range of health risks, both in humans and in animals. The association of filamentous fungi and their metabolites to different negative health conditions in humans and animals, has contributed to the importance of investigating different health risks induced by this family of heterotrophs. This review aims to discuss health risks associated with commonly occurring filamentous fungal species which belong to genera Aspergillus, Penicillium and Fusarium, as well as evaluating their pathogenicity and mycotoxic properties. PMID:28677641

  8. Biological nitrogen and phosphorus removal by filamentous bacteria ...

    African Journals Online (AJOL)

    Keywords: activated sludge, denitrification, glycogen accumulating organisms, filamentous bacteria, phosphorus removal. Introduction. Biological nutrient removal (BNR) has gained attention over chemical nutrient removal because of the high cost of the chemi- cal process and the large sludge volumes produced.

  9. Method for preparing metallated filament-wound structures

    Science.gov (United States)

    Peterson, George R.

    1979-01-01

    Metallated graphite filament-wound structures are prepared by coating a continuous multi-filament carbon yarn with a metal carbide, impregnating the carbide coated yarn with a polymerizable carbon precursor, winding the resulting filament about a mandrel, partially curing the impregnation in air, subjecting the wound composite to heat and pressure to cure the carbon precursor, and thereafter heating the composite in a sizing die at a pressure loading of at least 1000 psi for graphitizing the carbonaceous material in the composite. The carbide in the composite coalesces into rod-like shapes which are disposed in an end-to-end relationship parallel with the filaments to provide resistance to erosion in abrasive laden atmospheres.

  10. Positrusion Filament Recycling System for ISS, Phase II

    Data.gov (United States)

    National Aeronautics and Space Administration — The Positrusion ISS Recycler enables recycling of scrap and waste plastics into high-quality filament for 3D printers to enable sustainable in-situ manufacturing on...

  11. UHECR acceleration in dark matter filaments of cosmological structure formation

    Science.gov (United States)

    Malkov, M. A.; Sagdeev, R. Z.; Diamond, P. H.

    2011-04-01

    A mechanism for proton acceleration to ~ 1021 eV is suggested. It may operate in accretion flows onto thin dark matter filaments of cosmic structure formation. The flow compresses the ambient magnetic field to strongly increase and align it with the filament. Particles begin the acceleration by E × B drift with the accretion flow. The energy gain in the drift regime is limited by the conservation of the adiabatic invariant p⊥2/B(r). Upon approaching the filament, the drift turns into the gyro-motion around the filament so that the particle moves parallel to the azimuthal electric field. In this `betatron' regime the acceleration speeds up to rapidly reach the electrodynamic limit cpmax = eBR for an accelerator with magnetic field B and the orbit radius R (Larmor radius). The periodic orbit becomes unstable and the particle slings out of the filament to the region of a weak (uncompressed) magnetic field, which terminates the acceleration. To escape the filament, accelerated particles must have gyro-radii comparable with the filament radius. Therefore, the mechanism requires pre-acceleration that is likely to occur in large scale shocks upstream or nearby the filament accretion flow. Previous studies identify such shocks as efficient proton accelerators, with a firm upper limit ~ 1019.5 eV placed by the catastrophic photo-pion losses. The present mechanism combines explosive energy gain in its final (betatron) phase with prompt particle release from the region of strong magnetic field. It is this combination that allows protons to overcome both the photo-pion and the synchrotron-Compton losses and therefore attain energy ~ 1021 eV. A customary requirement on accelerator power to reach a given Emax, which is placed by the accelerator energy dissipation proptoEmax2/Z0 due to the finite vacuum impedance Z0, is circumvented by the cyclic operation of the accelerator.

  12. Impact of Submesoscale Processes on Dynamics of Phytoplankton Filaments

    Science.gov (United States)

    2015-02-12

    RESPONSIBLE PERSON 19b. TELEPHONE NUMBER (Include area code) 29-04-2015 Journal Article Impact of submesoscale processes on dynamics of phytoplankton ...in contrast to the earlier summer time, when the ASC mixes phytoplankton much deeper to the area below of the euphotic depth, and chlorophyll a...filaments are 3 -4 times weaker. coastal processes; upwelling, submesoscale processes, phytoplankton filaments Unclassified Unclassified Unclassified UU 13 Igor Shulman (228) 688-5646 Reset

  13. The architecture and fine structure of gill filaments in the brown ...

    African Journals Online (AJOL)

    Special attention was paid to filament architecture, ennervation of filaments, number and type of cells populating filament epithelia and variations in epithelial cell morphology and cilia ultrastructure. Filament shape was maintained by thickened chi-tln and strategically placed smooth myocytes. The epithelium was populated ...

  14. Treadmilling of actin filaments via Brownian dynamics simulations

    Science.gov (United States)

    Guo, Kunkun; Shillcock, Julian; Lipowsky, Reinhard

    2010-10-01

    Actin polymerization is coupled to the hydrolysis of adenosine triphosphate (ATP) into adenosine diphosphate (ADP) and inorganic phosphate (Pi). Therefore, each protomer within an actin filament can attain three different nucleotide states corresponding to bound ATP, ADP/Pi, and ADP. These protomer states form spatial patterns on the growing (or shrinking) filaments. Using Brownian dynamics simulations, the growth behavior of long filaments is studied, together with the associated protomer patterns, as a function of ATP-actin monomer concentration, CT, within the surrounding solution. For concentrations close to the critical concentration CT=CT,cr, the filaments undergo treadmilling, i.e., they grow at the barbed and shrink at the pointed end, which leads to directed translational motion of the whole filament. The corresponding nonequilibrium states are characterized by several global fluxes and by spatial density and flux profiles along the filaments. We focus on a certain set of transition rates as deduced from in vitro experiments and find that the associated treadmilling (or turnover) rate is about 0.08 monomers per second.

  15. The Weak Lensing Masses of Filaments between Luminous Red Galaxies

    Science.gov (United States)

    Epps, Seth D.; Hudson, Michael J.

    2017-07-01

    In the standard model of non-linear structure formation, a cosmic web of dark-matter-dominated filaments connects dark matter haloes. In this paper, we stack the weak lensing signal of an ensemble of filaments between groups and clusters of galaxies. Specifically, we detect the weak lensing signal, using CFHTLenS galaxy ellipticities, from stacked filaments between Sloan Digital Sky Survey (SDSS)-III/Baryon Oscillation Spectroscopic Survey luminous red galaxies (LRGs). As a control, we compare the physical LRG pairs with projected LRG pairs that are more widely separated in redshift space. We detect the excess filament mass density in the projected pairs at the 5σ level, finding a mass of (1.6 ± 0.3) × 1013 M⊙ for a stacked filament region 7.1 h-1 Mpc long and 2.5 h-1 Mpc wide. This filament signal is compared with a model based on the three-point galaxy-galaxy-convergence correlation function, as developed in Clampitt et al., yielding reasonable agreement.

  16. Design and optimize of 3-axis filament winding machine

    Science.gov (United States)

    Quanjin, Ma; Rejab, M. R. M.; Idris, M. S.; Bachtiar, B.; Siregar, J. P.; Harith, M. N.

    2017-10-01

    Filament winding technique is developed as the primary process for composite cylindrical structures fabrication at low cost. Fibres are wound on a rotating mandrel by a filament winding machine where resin impregnated fibres pass through a pay-out eye. This paper aims to develop and optimize a 3-axis, lightweight, practical, efficient, portable filament winding machine to satisfy the customer demand, which can fabricate pipes and round shape cylinders with resins. There are 3 main units on the 3-axis filament winding machine, which are the rotary unit, the delivery unit and control system unit. Comparison with previous existing filament winding machines in the factory, it has 3 degrees of freedom and can fabricate more complex shape specimens based on the mandrel shape and particular control system. The machine has been designed and fabricated on 3 axes movements with control system. The x-axis is for movement of the carriage, the y-axis is the rotation of mandrel and the z-axis is the movement of the pay-out eye. Cylindrical specimens with different dimensions and winding angles were produced. 3-axis automated filament winding machine has been successfully designed with simple control system.

  17. The electric toothbrush: analysis of filaments under stereomicroscope.

    Science.gov (United States)

    Checchi, L; Farina, E; Felice, P; Montevecchi, M

    2004-08-01

    The use of manual and electric toothbrushes has a fundamental role in primary prevention in oral hygiene. However, aggressive use of the toothbrush, especially those with non-rounded filaments, can result in lesions in both soft and hard oral tissue. Without doubt, the electric toothbrush is a useful aid for the patient, and it is therefore interesting to evaluate not only its effectiveness in plaque removal, but also the relationship between morphology of filaments and incidence of muco-gingival pathologies. The aim of this research was to evaluate various forms of bristles of electric toothbrushes under a stereomicroscope vision. Brushes tested included two samples of toothbrushes from six different types. Tufts from the same position on the toothbrush head were removed and examined under stereomicroscope. In this study the percentage of rounded filaments that is considered acceptable and non-traumatic was evaluated according to the Silverstone and Featherstone classification. Morphological analysis of electric toothbrush filaments revealed a low percentage of rounded filaments. In only four of 12 electric toothbrushes tested there were more than 50% of the filaments rounded in appearance.

  18. Effect of Filament Fineness on Composite Yarn Residual Torque

    Directory of Open Access Journals (Sweden)

    Sarıoğlu Esin

    2018-03-01

    Full Text Available Yarn residual torque or twist liveliness occurs when the twist is imparted to spin the fibers during yarn formation. It causes yarn snarling, which is an undesirable property and can lead the problems for further processes such as weaving and knitting. It affects the spirality of knitted fabrics and skewness of woven fabrics. Generally, yarn residual torque depends on yarn twist, yarn linear density, and fiber properties used. Composite yarns are widely produced to exploit two yarns with different properties such on optimum way at the same time and these yarns can be produced by wrapping sheath fibers around filament core fiber with a certain twist. In this study, the effect of filament fineness used as core component of composite yarn on residual torque was analyzed. Thus, the false twist textured polyester filament yarns with different filament fineness were used to produce composite yarns with different yarn count. The variance analysis was performed to determine the significance of twist liveliness of filament yarns and yarn count on yarn twist liveliness. Results showed that there is a statistically significant differences at significance level of α=0.05 between filament fineness and yarn residual torque of composite yarns.

  19. Optical and electrical properties of a spiral LED filament

    Science.gov (United States)

    Wang, Liping; Zou, Jun; Yang, Bobo; Li, Wenbo; Li, Yang; Shi, Mingming; Zhu, Wei; Zhang, Canyun; Wang, Fengchao; Lin, Yujie

    2018-02-01

    This paper introduces a new type of spiral white light-emitting diodes (WLED) filament with high luminous efficiency and uniform optical performance. The optical and thermal properties of the flexible filament were investigated at different stretching heights, namely 0, 1, 2, and 3 cm. The results indicated that the filament showed the best optical characteristics at the stretching height of 2 cm, because of good heat dissipation. In addition, the radiation temperature of the filament was inversely proportional to the output luminous flux. The reliability of the filament at a stretching height of 2 cm was also evaluated after 1000 h of use. The result demonstrated that the luminous flux decay of the bulb was only 0.85%. The flexible spiral WLED filament exhibiting high luminous flux and good reliability could be adapted to promote industrial development in the near future. Project supported by the National Nature Science Foundation of China (No. 51302171), the Science and Technology Commission of Shanghai Municipality (CN) (No. 14500503300), the Shanghai Municipal Alliance Program (No. Lm201547), the Shanghai Cooperative Project (No. ShanghaiCXY-2013-61), and the Jiashan County Technology Program (No. 20141316).

  20. Control of the phosphorylation of the astrocyte marker glial fibrillary acidic protein (GFAP in the immature rat hippocampus by glutamate and calcium ions: possible key factor in astrocytic plasticity

    Directory of Open Access Journals (Sweden)

    Rodnight R.

    1997-01-01

    Full Text Available The present review describes recent research on the regulation by glutamate and Ca2+ of the phosphorylation state of the intermediate filament protein of the astrocytic cytoskeleton, glial fibrillary acidic protein (GFAP, in immature hippocampal slices. The results of this research are discussed against a background of modern knowledge of the functional importance of astrocytes in the brain and of the structure and dynamic properties of intermediate filament proteins. Astrocytes are now recognized as partners with neurons in many aspects of brain function with important roles in neural plasticity. Site-specific phosphorylation of intermediate filament proteins, including GFAP, has been shown to regulate the dynamic equilibrium between the polymerized and depolymerized state of the filaments and to play a fundamental role in mitosis. Glutamate was found to increase the phosphorylation state of GFAP in hippocampal slices from rats in the post-natal age range of 12-16 days in a reaction that was dependent on external Ca2+. The lack of external Ca2+ in the absence of glutamate also increased GFAP phosphorylation to the same extent. These effects of glutamate and Ca2+ were absent in adult hippocampal slices, where the phosphorylation of GFAP was completely Ca2+-dependent. Studies using specific agonists of glutamate receptors showed that the glutamate response was mediated by a G protein-linked group II metabotropic glutamate receptor (mGluR. Since group II mGluRs do not act by liberating Ca2+ from internal stores, it is proposed that activation of the receptor by glutamate inhibits Ca2+ entry into the astrocytes and consequently down-regulates a Ca2+-dependent dephosphorylation cascade regulating the phosphorylation state of GFAP. The functional significance of these results may be related to the narrow developmental window when the glutamate response is present. In the rat brain this window corresponds to the period of massive synaptogenesis during

  1. Experimental study of infrared filaments under different initial conditions

    Science.gov (United States)

    Mirell, Daniel Joseph

    In 1964, four years after the first working laser was constructed, long skinny damage tracks and fluorescence trails were seen inside of certain transparent media that were excited by intense light pulses [1]. What was so remarkable about these features was the narrowness of the spatial profile and their long propagation length in the beam in concert with the very high intensity of the light that would be necessary to produce them. A purely linear model of light propagation through such media was insufficient to explain the results of these experiments and hence a new area of nonlinear optics, latex coined filamentation (to describe the length, slimness, and intensity of the light field), was born. Filament studies begin with a medium that has a nonlinear index of refraction, n¯2, that interacts with an intense beam of light so as to cause it to self-focus. The n¯2 of liquid and solid transparent media is much higher than the n¯ 2 of gases and therefore a much higher intensity of laser source would need to be invented to begin the study of filaments in air. With the advent of the Ti-Sapphire Kerr-lens modelocked laser [2], working in combination with the development of the chirped pulse amplifier system in the mid-1990's, light intensities sufficient to produce filaments in air was realized. Since that time much experimental and theoretical work has been done to better understand some of the additional complexities that arise specifically in the filamentation of light in air using several different wavelengths (UV to IR) and pulsewidths (femto- to pico-seconds). Many theoretical models exist each with a different emphasis on the various physical mechanisms that may produce the features experimentally observed in filaments. The experimental work has sought to give the theoretician better data on some of the properties of filaments such as the: (a) spatial and temporal structure of the beam and of the produced plasma (that arises due to the high intensity light

  2. Filamented plasmas in laser ablation of solids

    Science.gov (United States)

    Davies, J. R.; Fajardo, M.; Kozlová, M.; Mocek, T.; Polan, J.; Rus, B.

    2009-03-01

    We report results from laser-solid experiments at PALS using an x-ray laser probe with a pulse length of 0.1 ns and a wavelength of 21.2 nm. A laser with a pulse length of 0.3 ns, a peak intensity of up to 5 × 1013 W cm-2 and a wavelength of 1.3 µm was focused to a 0.15 mm wide line on 3 mm long zinc and 1 mm long iron targets and the probe was passed along the length of the plasma formed. The results show plasma 'hairs', or filaments, appearing only below the critical density, 0.1 ns before the peak of the laser pulse. The plasma around the critical density was clearly imaged and remained uniform. Magneto-hydrodynamic modelling indicates that this is caused by a magnetic field that diffuses from the critical surface, where it is generated, leading to a magnetic pressure comparable to the plasma pressure below the critical density. A dispersion relation is derived for density perturbations perpendicular to a temperature gradient in the presence of an existing magnetic field, which shows that such perturbations always grow, with the growth rate being the greatest for small wavelength perturbations and at low densities. These results indicate that the hair-like structures should be a typical feature of laser ablated plasmas below the critical density following significant plasma expansion, in agreement with numerous experimental results. The implications for x-ray lasers and fast ignition inertial confinement fusion are discussed.

  3. Methodologies and perspectives of proteomics applied to filamentous fungi: from sample preparation to secretome analysis.

    Science.gov (United States)

    Bianco, Linda; Perrotta, Gaetano

    2015-03-12

    Filamentous fungi possess the extraordinary ability to digest complex biomasses and mineralize numerous xenobiotics, as consequence of their aptitude to sensing the environment and regulating their intra and extra cellular proteins, producing drastic changes in proteome and secretome composition. Recent advancement in proteomic technologies offers an exciting opportunity to reveal the fluctuations of fungal proteins and enzymes, responsible for their metabolic adaptation to a large variety of environmental conditions. Here, an overview of the most commonly used proteomic strategies will be provided; this paper will range from sample preparation to gel-free and gel-based proteomics, discussing pros and cons of each mentioned state-of-the-art technique. The main focus will be kept on filamentous fungi. Due to the biotechnological relevance of lignocellulose degrading fungi, special attention will be finally given to their extracellular proteome, or secretome. Secreted proteins and enzymes will be discussed in relation to their involvement in bio-based processes, such as biomass deconstruction and mycoremediation.

  4. Self-assembly of designed supramolecular magnetic filaments of different shapes

    Energy Technology Data Exchange (ETDEWEB)

    Novak, E.V. [Ural Federal University, Lenin Av. 51, Ekaterinburg (Russian Federation); Rozhkov, D.A., E-mail: d.a.rozhkov@gmail.com [Ural Federal University, Lenin Av. 51, Ekaterinburg (Russian Federation); Sanchez, P.A. [University of Vienna, Sensengasse 8, Vienna (Austria); Kantorovich, S.S. [Ural Federal University, Lenin Av. 51, Ekaterinburg (Russian Federation); University of Vienna, Sensengasse 8, Vienna (Austria)

    2017-06-01

    In the present work we study via molecular dynamics simulations filaments of ring and linear shape. Filaments are made of magnetic nanoparticles, possessing a point dipole in their centres. Particles in filaments are crosslinked in a particular way, so that the deviation of the neighbouring dipoles from the head-to-tail orientation is penalised by the bond. We show how the conformation of a single chain and ring filament changes on cooling for different lengths. We also study filament pairs, by fixing filaments at a certain distance and analysing the impact of inter-filament interaction on the equilibrium configurations. Our study opens a perspective to investigate the dispersions of filaments, both theoretically and numerically, by using effective potentials. - Highlights: • Single filament study. • Magnetic particles crosslinked in chains and rings. • Magnetic filament interactions.

  5. Giant quiescent solar filament observed with high-resolution spectroscopy

    Science.gov (United States)

    Kuckein, C.; Verma, M.; Denker, C.

    2016-05-01

    Aims: An extremely large filament was studied in various layers of the solar atmosphere. The inferred physical parameters and the morphological aspects are compared with smaller quiescent filaments. Methods: A giant quiet-Sun filament was observed with the high-resolution Echelle spectrograph at the Vacuum Tower Telescope at Observatorio del Teide, Tenerife, Spain, on 2011 November 15. A mosaic of spectra (ten maps of 100″ × 182″) was recorded simultaneously in the chromospheric absorption lines Hα and Na I D2. Physical parameters of the filament plasma were derived using cloud model (CM) inversions and line core fits. The spectra were complemented with full-disk filtergrams (He I λ10830 Å, Hα, and Ca II K) of the Chromospheric Telescope (ChroTel) and full-disk magnetograms of the Helioseismic and Magnetic Imager (HMI). Results: The filament had extremely large linear dimensions (~817 arcsec), which corresponds to about 658 Mm along a great circle on the solar surface. A total amount of 175119 Hα contrast profiles were inverted using the CM approach. The inferred mean line-of-sight (LOS) velocity, Doppler width, and source function were similar to previous works of smaller quiescent filaments. However, the derived optical thickness was higher. LOS velocity trends inferred from the Hα line core fits were in accord but weaker than those obtained with CM inversions. Signatures of counter-streaming flows were detected in the filament. The largest brightening conglomerates in the line core of Na I D2 coincided well with small-scale magnetic fields as seen by HMI. Mixed magnetic polarities were detected close to the ends of barbs. The computation of photospheric horizontal flows based on HMI magnetograms revealed flow kernels with a size of 5-8 Mm and velocities of 0.30-0.45 km s-1 at the ends of the filament. Conclusions: The physical properties of extremely large filaments are similar to their smaller counterparts, except for the optical thickness, which in

  6. Deinococcus radiodurans RecA nucleoprotein filaments characterized at the single-molecule level with optical tweezers

    Energy Technology Data Exchange (ETDEWEB)

    Pobegalov, Georgii, E-mail: george.pobegalov@nanobio.spbstu.ru [Peter the Great St.Petersburg Polytechnic University, Saint-Petersburg 195251 (Russian Federation); Cherevatenko, Galina; Alekseev, Aleksandr; Sabantsev, Anton; Kovaleva, Oksana; Vedyaykin, Alexey; Morozova, Natalia [Peter the Great St.Petersburg Polytechnic University, Saint-Petersburg 195251 (Russian Federation); Baitin, Dmitrii [Peter the Great St.Petersburg Polytechnic University, Saint-Petersburg 195251 (Russian Federation); Petersburg Nuclear Physics Institute, NRC Kurchatov Institute, Gatchina 188300 (Russian Federation); Khodorkovskii, Mikhail [Peter the Great St.Petersburg Polytechnic University, Saint-Petersburg 195251 (Russian Federation)

    2015-10-23

    Deinococcus radiodurans can survive extreme doses of ionizing radiation due to the very efficient DNA repair mechanisms that are able to cope even with hundreds of double-strand breaks. RecA, the critical protein of homologous recombination in bacteria, is one of the key components of the DNA-repair system. Repair of double-strand breaks requires RecA binding to DNA and assembly of the RecA nucleoprotein helical filaments. The Escherichia coli RecA protein (EcRecA) and its interactions with DNA have been extensively studied using various approaches including single-molecule techniques, while the D. radiodurans RecA (DrRecA) remains much less characterized. However, DrRecA shows some remarkable differences from E. coli homolog. Here we combine microfluidics and single-molecule DNA manipulation with optical tweezers to follow the binding of DrRecA to long double-stranded DNA molecules and probe the mechanical properties of DrRecA nucleoprotein filaments at physiological pH. Our data provide a direct comparison of DrRecA and EcRecA binding to double-stranded DNA under identical conditions. We report a significantly faster filaments assembly as well as lower values of persistence length and contour length for DrRecA nucleoprotein filaments compared to EcRecA. Our results support the existing model of DrRecA forming more frequent and less continuous filaments relative to those of EcRecA. - Highlights: • We investigate Deinococcus radiodurans RecA interactions with long double-stranded DNA at the single-molecule level. • At physiological pH D. radiodurans RecA forms nucleoprotein filaments significantly faster relative to Escherichia coli RecA. • D. radiodurans RecA-dsDNA nucleoprotein filaments are more flexible and slightly shorter compared to those of E. coli RecA.

  7. Electrostatic interactions between the Bni1p Formin FH2 domain and actin influence actin filament nucleation.

    Science.gov (United States)

    Baker, Joseph L; Courtemanche, Naomi; Parton, Daniel L; McCullagh, Martin; Pollard, Thomas D; Voth, Gregory A

    2015-01-06

    Formins catalyze nucleation and growth of actin filaments. Here, we study the structure and interactions of actin with the FH2 domain of budding yeast formin Bni1p. We built an all-atom model of the formin dimer on an Oda actin filament 7-mer and studied structural relaxation and interprotein interactions by molecular dynamics simulations. These simulations produced a refined model for the FH2 dimer associated with the barbed end of the filament and showed electrostatic interactions between the formin knob and actin target-binding cleft. Mutations of two formin residues contributing to these interactions (R1423N, K1467L, or both) reduced the interaction energies between the proteins, and in coarse-grained simulations, the formin lost more interprotein contacts with an actin dimer than with an actin 7-mer. Biochemical experiments confirmed a strong influence of these mutations on Bni1p-mediated actin filament nucleation, but not elongation, suggesting that different interactions contribute to these two functions of formins. Copyright © 2015 Elsevier Ltd. All rights reserved.

  8. Mechanistic Insights Into Filamentous Phage Integration In Vibrio cholerae

    Directory of Open Access Journals (Sweden)

    Bhabatosh eDas

    2014-11-01

    Full Text Available Vibrio cholerae, the etiological agent of acute diarrhoeal disease cholera, harbors large numbers of lysogenic filamentous phages, contribute significantly to the host pathogenesis and provide fitness factors to the pathogen that help the bacterium to survive in natural environment. Most of the vibriophage genomes are not equipped with integrase and thus exploit two host-encoded tyrosine recombinases, XerC and XerD, for lysogenic conversion. Integration is site-specific and it occurs at dimer resolution site (dif of either one or both chromosomes of V. cholerae. Each dif sequence contains two recombinase-binding sequences flanking a central region. The integration follows a sequential strand exchanges between dif and attP sites within a DNA-protein complex consisting of one pair of each recombinase and two DNA fragments. During entire process of recombination, both the DNA components and recombinases of the synaptic complex keep transiently interconnected. Within the context of synaptic complex, both of the actuated enzymes mediate cleavage of phosphodiester bonds. First cleavage generates a phosphotyrosyl-linked recombinase-DNA complex at the recombinase binding sequence and free 5’-hydroxyl end at the first base of the central region. Following the cleavage, the exposed bases with 5’-hydroxyl ends of the central region of dif and attP sites melt from their complementary strands and react with the recombinase-DNA phosphotyrosyl linkage of their recombining partner. Subsequent ligation between dif and attP strands requires complementary base pair interactions at the site of phosphodiester bond formation. Integration mechanism is mostly influenced by the compatibility of dif and attP sequences. dif sites are highly conserved across bacterial phyla. Different phage genomes have different attP sequences; therefore they rely on different mechanisms for integration. Here, I review our current understanding of integration mechanisms used by the

  9. Modified Filamentous Bacteriophage as a Scaffold for Carbon Nanofiber.

    Science.gov (United States)

    Szot-Karpińska, Katarzyna; Golec, Piotr; Leśniewski, Adam; Pałys, Barbara; Marken, Frank; Niedziółka-Jönsson, Joanna; Węgrzyn, Grzegorz; Łoś, Marcin

    2016-12-21

    With the advent of nanotechnology, carbon nanomaterials such as carbon nanofibers (CNF) have aroused substantial interest in various research fields, including energy storage and sensing. Further improvement of their properties might be achieved via the application of viral particles such as bacteriophages. In this report, we present a filamentous M13 bacteriophage with a point mutation in gene VII (pVII-mutant-M13) that selectively binds to the carbon nanofibers to form 3D structures. The phage-display technique was utilized for the selection of the pVII-mutant-M13 phage from the phage display peptide library. The properties of this phage make it a prospective candidate for a scaffold material for CNFs. The results for binding of CNF by mutant phage were compared with those for maternal bacteriophage (pVII-M13). The efficiency of binding between pVII-mutant-M13 and CNF is about 2 orders of magnitude higher compared to that of the pVII-M13. Binding affinity between pVII-mutant-M13 and CNF was also characterized using atomic force microscopy, scanning electron microscopy, and transmission electron microscopy, which confirmed the specificity of the interaction of the phage pVII-mutant-M13 and the CNF; the binding occurs via the phage's ending, where the mutated pVII protein is located. No similar behavior has been observed for other carbon nanomaterials such as graphite, reduced graphene oxide, single-walled carbon nanotubes, and multiwalled carbon nanotubes. Infrared spectra confirmed differences in the interaction with CNF between the pVII-mutant-M13 and the pVII-M13. Basing on conducted research, we hypothesize that the interactions are noncovalent in nature, with π-π interactions playing the dominant role. Herein, the new bioconjugate material is introduced.

  10. Large-amplitude Longitudinal Oscillations in a Solar Filament

    Science.gov (United States)

    Zhang, Q. M.; Li, T.; Zheng, R. S.; Su, Y. N.; Ji, H. S.

    2017-06-01

    In this paper, we report our multiwavelength observations of the large-amplitude longitudinal oscillations of a filament observed on 2015 May 3. Located next to active region 12335, the sigmoidal filament was observed by the ground-based Hα telescopes from the Global Oscillation Network Group and by the Atmospheric Imaging Assembly instrument on board the Solar Dynamics Observatory. The filament oscillations were most probably triggered by the magnetic reconnection in the filament channel, which is characterized by the bidirectional flows, brightenings in EUV and soft X-ray, and magnetic cancellation in the photosphere. The directions of oscillations have angles of 4°-36° with respect to the filament axis. The whole filament did not oscillate in phase as a rigid body. Meanwhile, the oscillation periods (3100-4400 s) have a spatial dependence, implying that the curvature radii (R) of the magnetic dips are different at different positions. The values of R are estimated to be 69.4-133.9 Mm, and the minimum transverse magnetic field of the dips is estimated to be 15 G. The amplitudes of S5-S8 grew with time, while the amplitudes of S9-S14 damped with time. The oscillation amplitudes range from a few to ten Mm, and the maximum velocity can reach 30 km s-1. Interestingly, the filament experienced mass drainage southward at a speed of ˜27 km s-1. The oscillations continued after the mass drainage and lasted for more than 11 hr. After the mass drainage, the oscillation phases did not change much. The periods of S5-S8 decreased, while the periods of S9-S14 increased. The amplitudes of S5-S8 damped with time, while the amplitudes of S9-S14 grew. Most of the damping (growing) ratios are between -9 and 14. We offer a schematic cartoon to explain the complex behaviors of oscillations by introducing thread-thread interaction.

  11. Closely spaced fine filament multifilamentary NbTi strands

    International Nuclear Information System (INIS)

    Gregory, E.; Liu, H.; Seuntjens, J.M.

    1994-01-01

    A series of papers showing the advantages of close spacing and matrix alloying for the development of high J c , fine filament, NbTi materials which have low electrical coupling have appeared in the last seven years. In order to achieve the highest J c 's, it has been shown that close spacing has many advantages. This, however, leads to proximity coupling which has to be overcome by the addition of alloying elements to the matrix between the filaments. Of the three alloying materials normally used for this purpose, Ni, Si, and Mn, the most effective is Mn, which operates by a spin flip scattering mechanism whereas Ni and Si produce decoupling by less effective resistive scattering. Ni and Si, however, harden the matrix more than does the small amount of Mn, [0.5wt%], which has been used in most of the past work on the reduction of proximity coupling. This hardening allows the filaments to be separated to a greater extent than is possible in the case of a pure copper matrix without a significant increase in filament sausaging and a resultant J c decrease. Silicon also has one additional advantage over the other alloying elements in that it reduces the formation of compounds on the surface of the filaments, thus it may obviate the necessity for a Nb barrier layer and thus allow an even greater increase in J c . In this paper the authors explore further some of the effects of the addition of manganese and/or silicon to the matrix between the filaments in an effort to optimize properties at the smaller filament sizes

  12. A general insert label for peptide display on chimeric filamentous bacteriophages.

    Science.gov (United States)

    Kaplan, Gilad; Gershoni, Jonathan M

    2012-01-01

    The foreign insert intended to be displayed via recombinant phage proteins can have a negative effect on protein expression and phage assembly. A typical example is the case of display of peptides longer than 6 amino acid residues on the major coat protein, protein VIII of the filamentous bacteriophages M13 and fd. A solution to this problem has been the use of "two-gene systems" generating chimeric phages that concomitantly express wild-type protein VIII along with recombinant protein VIII. Although the two-gene systems are much more permissive in regard to insert length and composition, some cases can still adversely affect phage assembly. Although these phages genotypically contain the desired DNA of the insert, they appear to be phenotypically wild type. To avoid false-negative results when using chimeric phages in binding studies, it is necessary to confirm that the observed lack of phage recognition is not due to faulty assembly and display of the intended insert. Here we describe a strategy for generating antibodies that specifically recognize recombinant protein VIII regardless of the nature of its foreign insert. These antibodies can be used as a general monitor of the display of recombinant protein VIII into phage particles. Copyright © 2011 Elsevier Inc. All rights reserved.

  13. Large-amplitude Longitudinal Oscillations Triggered by the Merging of Two Solar Filaments: Observations and Magnetic Field Analysis

    Science.gov (United States)

    Luna, M.; Su, Y.; Schmieder, B.; Chandra, R.; Kucera, T. A.

    2017-12-01

    We follow the eruption of two related intermediate filaments observed in Hα (from GONG) and EUV (from Solar Dynamics Observatory SDO/Atmospheric Imaging assembly AIA) and the resulting large-amplitude longitudinal oscillations of the plasma in the filament channels. The events occurred in and around the decayed active region AR12486 on 2016 January 26. Our detailed study of the oscillation reveals that the periods of the oscillations are about one hour. In Hα, the period decreases with time and exhibits strong damping. The analysis of 171 Å images shows that the oscillation has two phases: an initial long-period phase and a subsequent oscillation with a shorter period. In this wavelength, the damping appears weaker than in Hα. The velocity is the largest ever detected in a prominence oscillation, approximately 100 {km} {{{s}}}-1. Using SDO/HMI magnetograms, we reconstruct the magnetic field of the filaments, modeled as flux ropes by using a flux-rope insertion method. Applying seismological techniques, we determine that the radii of curvature of the field lines in which cool plasma is condensed are in the range 75-120 Mm, in agreement with the reconstructed field. In addition, we infer a field strength of ≥7 to 30 Gauss, depending on the electron density assumed, that is also in agreement with the values from the reconstruction (8-20 Gauss). The poloidal flux is zero and the axis flux is on the order of 1020 to 1021 Mx, confirming the high shear existing even in a non-active filament.

  14. The costa of trichomonads: A complex macromolecular cytoskeleton structure made of uncommon proteins.

    Science.gov (United States)

    de Andrade Rosa, Ivone; Caruso, Marjolly Brigido; de Oliveira Santos, Eidy; Gonzaga, Luiz; Zingali, Russolina Benedeta; de Vasconcelos, Ana Tereza R; de Souza, Wanderley; Benchimol, Marlene

    2017-06-01

    The costa is a prominent striated fibre that is found in protozoa of the Trichomonadidae family that present an undulating membrane. It is composed primarily of proteins that have not yet been explored. In this study, we used cell fractionation to obtain a highly enriched costa fraction whose structure and composition was further analysed by electron microscopy and mass spectrometry. Electron microscopy of negatively stained samples revealed that the costa, which is a periodic structure with alternating electron-dense and electron-lucent bands, displays three distinct regions, named the head, neck and body. Fourier transform analysis showed that the electron-lucent bands present sub-bands with a regular pattern. An analysis of the costa fraction via one- and two-dimensional electrophoresis and liquid chromatography coupled with tandem mass spectrometry (LC-MS/MS) allowed the identification of 54 hypothetical proteins. Fourteen of those proteins were considered to be major components of the fraction. The costa of T. foetus is a complex and organised cytoskeleton structure made of a large number of proteins which is assembled into filamentous structures. Some of these proteins exhibit uncharacterised domains and no function related according to gene ontology, suggesting that the costa structure may be formed by a new class of proteins that differ from those previously described in other organisms. Seven of these proteins contain prefoldin domains displaying coiled-coil regions. This propriety is shared with proteins of the striated fibres of other protozoan as well as in intermediate filaments. Our observations suggest the presence of a new class of the cytoskeleton filaments in T. foetus. We believe that our data could auxiliate in determining the specific locations of these proteins in the distinct regions that compose the costa, as well as to define the functional roles of each component. Therefore, our study will help in the better understanding of the

  15. Hydrophilins in the filamentous fungus Neosartorya fischeri (Aspergillus fischeri) have protective activity against several types of microbial water stress.

    Science.gov (United States)

    van Leeuwen, M R; Wyatt, T T; van Doorn, T M; Lugones, L G; Wösten, H A B; Dijksterhuis, J

    2016-02-01

    Hydrophilins are proteins that occur in all domains of life and protect cells and organisms against drought and other stresses. They include most of the late embryogenesis abundant (LEA) proteins and the heat shock protein (HSP) Hsp12. Here, the role of a predicted LEA-like protein (LeamA) and two Hsp12 proteins (Hsp12A and Hsp12B) of Neosartorya fischeri was studied. This filamentous fungus forms ascospores that belong to the most stress-resistant eukaryotic cells described to date. Heterologous expression of LeamA, Hsp12A and Hsp12B resulted in increased tolerance against salt and osmotic stress in Escherichia coli. These proteins were also shown to protect lactate dehydrogenase against dry heat and freeze-thaw cycles in vitro. Deletion of leamA caused diminished viability of sexual ascospores after drought and heat. This is the first report on functionality of Hsp12 and putative LeamA proteins derived from filamentous fungi, and their possible role in N. fischeri ascospore resistance against desiccation, high temperature and osmotic stress is discussed. © 2015 Society for Applied Microbiology and John Wiley & Sons Ltd.

  16. Magnetic Fields in the Massive Dense Cores of the DR21 Filament: Weakly Magnetized Cores in a Strongly Magnetized Filament

    Energy Technology Data Exchange (ETDEWEB)

    Ching, Tao-Chung; Lai, Shih-Ping [Institute of Astronomy and Department of Physics, National Tsing Hua University, Hsinchu 30013, Taiwan (China); Zhang, Qizhou; Girart, Josep M. [Harvard-Smithsonian Center for Astrophysics, 60 Garden Street, Cambridge MA 02138 (United States); Qiu, Keping [School of Astronomy and Space Science, Nanjing University, 163 Xianlin Avenue, Nanjing 210023 (China); Liu, Hauyu B., E-mail: chingtaochung@gmail.com [European Southern Observatory (ESO), Karl-Schwarzschild-Str. 2, D-85748 Garching (Germany)

    2017-04-01

    We present Submillimeter Array 880 μ m dust polarization observations of six massive dense cores in the DR21 filament. The dust polarization shows complex magnetic field structures in the massive dense cores with sizes of 0.1 pc, in contrast to the ordered magnetic fields of the parsec-scale filament. The major axes of the massive dense cores appear to be aligned either parallel or perpendicular to the magnetic fields of the filament, indicating that the parsec-scale magnetic fields play an important role in the formation of the massive dense cores. However, the correlation between the major axes of the cores and the magnetic fields of the cores is less significant, suggesting that during the core formation, the magnetic fields below 0.1 pc scales become less important than the magnetic fields above 0.1 pc scales in supporting a core against gravity. Our analysis of the angular dispersion functions of the observed polarization segments yields a plane-of-sky magnetic field strength of 0.4–1.7 mG for the massive dense cores. We estimate the kinematic, magnetic, and gravitational virial parameters of the filament and the cores. The virial parameters show that the gravitational energy in the filament dominates magnetic and kinematic energies, while the kinematic energy dominates in the cores. Our work suggests that although magnetic fields may play an important role in a collapsing filament, the kinematics arising from gravitational collapse must become more important than magnetic fields during the evolution from filaments to massive dense cores.

  17. A comparison study of a solar active-region eruptive filament and a neighboring non-eruptive filament

    International Nuclear Information System (INIS)

    Jiang, Chao-Wei; Feng, Xue-Shang; Wu, Shi-Tsan; Hu, Qiang

    2016-01-01

    Solar active region (AR) 11283 is a very magnetically complex region and it has produced many eruptions. However, there exists a non-eruptive filament in the plage region just next to an eruptive one in the AR, which gives us an opportunity to perform a comparison analysis of these two filaments. The coronal magnetic field extrapolated using our CESE–MHD–NLFFF code reveals that two magnetic flux ropes (MFRs) exist in the same extrapolation box supporting these two filaments, respectively. Analysis of the magnetic field shows that the eruptive MFR contains a bald-patch separatrix surface (BPSS) cospatial very well with a pre-eruptive EUV sigmoid, which is consistent with the BPSS model for coronal sigmoids. The magnetic dips of the non-eruptive MFRs match Hα observation of the non-eruptive filament strikingly well, which strongly supports the MFR-dip model for filaments. Compared with the non-eruptive MFR/filament (with a length of about 200 Mm), the eruptive MFR/filament is much smaller (with a length of about 20 Mm), but it contains most of the magnetic free energy in the extrapolation box and holds a much higher free energy density than the non-eruptive one. Both the MFRs are weakly twisted and cannot trigger kink instability. The AR eruptive MFR is unstable because its axis reaches above a critical height for torus instability, at which the overlying closed arcades can no longer confine the MFR stably. On the contrary, the quiescent MFR is very firmly held by its overlying field, as its axis apex is far below the torus-instability threshold height. Overall, this comparison investigation supports that an MFR can exist prior to eruption and the ideal MHD instability can trigger an MFR eruption. (paper)

  18. A comparison study of a solar active-region eruptive filament and a neighboring non-eruptive filament

    Science.gov (United States)

    Jiang, Chao-Wei; Wu, Shi-Tsan; Feng, Xue-Shang; Hu, Qiang

    2016-01-01

    Solar active region (AR) 11283 is a very magnetically complex region and it has produced many eruptions. However, there exists a non-eruptive filament in the plage region just next to an eruptive one in the AR, which gives us an opportunity to perform a comparison analysis of these two filaments. The coronal magnetic field extrapolated using our CESE-MHD-NLFFF code reveals that two magnetic flux ropes (MFRs) exist in the same extrapolation box supporting these two filaments, respectively. Analysis of the magnetic field shows that the eruptive MFR contains a bald-patch separatrix surface (BPSS) cospatial very well with a pre-eruptive EUV sigmoid, which is consistent with the BPSS model for coronal sigmoids. The magnetic dips of the non-eruptive MFRs match Hα observation of the non-eruptive filament strikingly well, which strongly supports the MFR-dip model for filaments. Compared with the non-eruptive MFR/filament (with a length of about 200 Mm), the eruptive MFR/filament is much smaller (with a length of about 20 Mm), but it contains most of the magnetic free energy in the extrapolation box and holds a much higher free energy density than the non-eruptive one. Both the MFRs are weakly twisted and cannot trigger kink instability. The AR eruptive MFR is unstable because its axis reaches above a critical height for torus instability, at which the overlying closed arcades can no longer confine the MFR stably. On the contrary, the quiescent MFR is very firmly held by its overlying field, as its axis apex is far below the torus-instability threshold height. Overall, this comparison investigation supports that an MFR can exist prior to eruption and the ideal MHD instability can trigger an MFR eruption.

  19. Rapid Formation and Disappearance of a Filament Barb

    Science.gov (United States)

    Joshi, Anand D.; Srivastava, Nandita; Mathew, Shibu K.; Martin, Sara F.

    2013-11-01

    We present observations of an activated quiescent filament obtained in Hα from the high-resolution Dutch Open Telescope (DOT) on 20 August 2010. The filament developed a barb in 10 min, which disappeared within the next 35 min. A data set from the DOT spanning 2 h was used to analyse this event. Line-of-sight velocity maps were constructed from the Doppler images, which reveal flows in filament spine during this period. Photospheric magnetograms were used from the Helioseismic and Magnetic Imager (HMI) on board the Solar Dynamics Observatory (SDO) to determine the changes in magnetic flux in the region surrounding the barb location. The analysis shows flows in the filament spine towards the barb location preceding its formation, and flows in the barb towards the spine during its disappearance. Magnetograms reveal patches of minority polarity flux close to the end of the barb at its greatest elongation. The flows in the spine and barbs are along numerous threads that compose these typical filament structures. The flows are consistent with field-aligned threads and demonstrate that the replacement time of the mass in barbs, and by inference, in the spine is very rapid.

  20. High-Resolution Observations of a Filament showing Activated Barb

    Science.gov (United States)

    Joshi, Anand; Martin, Sara F.; Mathew, Shibu; Srivastava, Nandita

    2012-07-01

    Analysis of a