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Sample records for nuclear envelope breakdown

  1. Nuclear envelope breakdown induced by herpes simplex virus type 1 involves the activity of viral fusion proteins.

    PubMed

    Maric, Martina; Haugo, Alison C; Dauer, William; Johnson, David; Roller, Richard J

    2014-07-01

    Herpesvirus infection reorganizes components of the nuclear lamina usually without loss of integrity of the nuclear membranes. We report that wild-type HSV infection can cause dissolution of the nuclear envelope in transformed mouse embryonic fibroblasts that do not express torsinA. Nuclear envelope breakdown is accompanied by an eight-fold inhibition of virus replication. Breakdown of the membrane is much more limited during infection with viruses that lack the gB and gH genes, suggesting that breakdown involves factors that promote fusion at the nuclear membrane. Nuclear envelope breakdown is also inhibited during infection with virus that does not express UL34, but is enhanced when the US3 gene is deleted, suggesting that envelope breakdown may be enhanced by nuclear lamina disruption. Nuclear envelope breakdown cannot compensate for deletion of the UL34 gene suggesting that mixing of nuclear and cytoplasmic contents is insufficient to bypass loss of the normal nuclear egress pathway.

  2. A novel mechanism of nuclear envelope break-down in a fungus: nuclear migration strips off the envelope

    PubMed Central

    Straube, Anne; Weber, Isabella; Steinberg, Gero

    2005-01-01

    In animals, the nuclear envelope disassembles in mitosis, while budding and fission yeast form an intranuclear spindle. Ultrastructural data indicate that basidiomycetes, such as the pathogen Ustilago maydis, undergo an ‘open mitosis'. Here we describe the mechanism of nuclear envelope break-down in U. maydis. In interphase, the nucleus resides in the mother cell and the spindle pole body is inactive. Prior to mitosis, it becomes activated and nucleates microtubules that reach into the daughter cell. Dynein appears at microtubule tips and exerts force on the spindle pole body, which leads to the formation of a long nuclear extension that reaches into the bud. Chromosomes migrate through this extension and together with the spindle pole bodies leave the old envelope, which remains in the mother cell until late telophase. Inhibition of nuclear migration or deletion of a Tem1p-like GTPase leads to a ‘closed' mitosis, indicating that spindle pole bodies have to reach into the bud where MEN signalling participates in envelope removal. Our data indicate that dynein-mediated premitotic nuclear migration is essential for envelope removal in U. maydis. PMID:15861140

  3. Nuclear envelope breakdown induced by herpes simplex virus type 1 involves the activity of viral fusion proteins

    SciTech Connect

    Maric, Martina; Haugo, Alison C.; Dauer, William; Johnson, David; Roller, Richard J.

    2014-07-15

    Herpesvirus infection reorganizes components of the nuclear lamina usually without loss of integrity of the nuclear membranes. We report that wild-type HSV infection can cause dissolution of the nuclear envelope in transformed mouse embryonic fibroblasts that do not express torsinA. Nuclear envelope breakdown is accompanied by an eight-fold inhibition of virus replication. Breakdown of the membrane is much more limited during infection with viruses that lack the gB and gH genes, suggesting that breakdown involves factors that promote fusion at the nuclear membrane. Nuclear envelope breakdown is also inhibited during infection with virus that does not express UL34, but is enhanced when the US3 gene is deleted, suggesting that envelope breakdown may be enhanced by nuclear lamina disruption. Nuclear envelope breakdown cannot compensate for deletion of the UL34 gene suggesting that mixing of nuclear and cytoplasmic contents is insufficient to bypass loss of the normal nuclear egress pathway. - Highlights: • We show that wild-type HSV can induce breakdown of the nuclear envelope in a specific cell system. • The viral fusion proteins gB and gH are required for induction of nuclear envelope breakdown. • Nuclear envelope breakdown cannot compensate for deletion of the HSV UL34 gene.

  4. Parvoviruses Cause Nuclear Envelope Breakdown by Activating Key Enzymes of Mitosis

    PubMed Central

    Porwal, Manvi; Cohen, Sarah; Snoussi, Kenza; Popa-Wagner, Ruth; Anderson, Fenja; Dugot-Senant, Nathalie; Wodrich, Harald; Dinsart, Christiane; Kleinschmidt, Jürgen A.; Panté, Nelly; Kann, Michael

    2013-01-01

    Disassembly of the nuclear lamina is essential in mitosis and apoptosis requiring multiple coordinated enzymatic activities in nucleus and cytoplasm. Activation and coordination of the different activities is poorly understood and moreover complicated as some factors translocate between cytoplasm and nucleus in preparatory phases. Here we used the ability of parvoviruses to induce nuclear membrane breakdown to understand the triggers of key mitotic enzymes. Nuclear envelope disintegration was shown upon infection, microinjection but also upon their application to permeabilized cells. The latter technique also showed that nuclear envelope disintegration was independent upon soluble cytoplasmic factors. Using time-lapse microscopy, we observed that nuclear disassembly exhibited mitosis-like kinetics and occurred suddenly, implying a catastrophic event irrespective of cell- or type of parvovirus used. Analyzing the order of the processes allowed us to propose a model starting with direct binding of parvoviruses to distinct proteins of the nuclear pore causing structural rearrangement of the parvoviruses. The resulting exposure of domains comprising amphipathic helices was required for nuclear envelope disintegration, which comprised disruption of inner and outer nuclear membrane as shown by electron microscopy. Consistent with Ca++ efflux from the lumen between inner and outer nuclear membrane we found that Ca++ was essential for nuclear disassembly by activating PKC. PKC activation then triggered activation of cdk-2, which became further activated by caspase-3. Collectively our study shows a unique interaction of a virus with the nuclear envelope, provides evidence that a nuclear pool of executing enzymes is sufficient for nuclear disassembly in quiescent cells, and demonstrates that nuclear disassembly can be uncoupled from initial phases of mitosis. PMID:24204256

  5. Virtual Nuclear Envelope Breakdown and Its Regulators in Fission Yeast Meiosis

    PubMed Central

    Asakawa, Haruhiko; Yang, Hui-Ju; Hiraoka, Yasushi; Haraguchi, Tokuko

    2016-01-01

    Ran, a small GTPase, is required for the spindle formation and nuclear envelope (NE) formation. After NE breakdown (NEBD) during mitosis in metazoan cells, the Ran-GTP gradient across the NE is lost and Ran-GTP becomes concentrated around chromatin, thus affecting the stability of microtubules and promoting the assembly of spindle microtubules and segregation of chromosomes. Mitosis in which chromosomes are segregated subsequent to NEBD is called “open mitosis.” In contrast, many fungi undergo a process termed “closed mitosis” in which chromosome segregation and spindle formation occur without NEBD. Although the fission yeast Schizosaccharomyces pombe undergoes a closed mitosis, it exhibits a short period during meiosis (anaphase of the second meiosis; called “anaphase II”) when nuclear and cytoplasmic proteins are mixed in the presence of intact NE and nuclear pore complexes (NPC). This “virtual” nuclear envelope breakdown (vNEBD) involves changes in the localization of RanGAP1, an activator of Ran-GTP hydrolysis. Recently, Nup132, a component of the structural core Nup107-160 subcomplex of the NPC, has been shown to be involved in the maintenance of the nuclear cytoplasmic barrier in yeast meiosis. In this review, we highlight the possible roles of RanGAP1 and Nup132 in vNEBD and discuss the biological significance of vNEBD in S. pombe meiosis. PMID:26870731

  6. Virtual Nuclear Envelope Breakdown and Its Regulators in Fission Yeast Meiosis.

    PubMed

    Asakawa, Haruhiko; Yang, Hui-Ju; Hiraoka, Yasushi; Haraguchi, Tokuko

    2016-01-01

    Ran, a small GTPase, is required for the spindle formation and nuclear envelope (NE) formation. After NE breakdown (NEBD) during mitosis in metazoan cells, the Ran-GTP gradient across the NE is lost and Ran-GTP becomes concentrated around chromatin, thus affecting the stability of microtubules and promoting the assembly of spindle microtubules and segregation of chromosomes. Mitosis in which chromosomes are segregated subsequent to NEBD is called "open mitosis." In contrast, many fungi undergo a process termed "closed mitosis" in which chromosome segregation and spindle formation occur without NEBD. Although the fission yeast Schizosaccharomyces pombe undergoes a closed mitosis, it exhibits a short period during meiosis (anaphase of the second meiosis; called "anaphase II") when nuclear and cytoplasmic proteins are mixed in the presence of intact NE and nuclear pore complexes (NPC). This "virtual" nuclear envelope breakdown (vNEBD) involves changes in the localization of RanGAP1, an activator of Ran-GTP hydrolysis. Recently, Nup132, a component of the structural core Nup107-160 subcomplex of the NPC, has been shown to be involved in the maintenance of the nuclear cytoplasmic barrier in yeast meiosis. In this review, we highlight the possible roles of RanGAP1 and Nup132 in vNEBD and discuss the biological significance of vNEBD in S. pombe meiosis. PMID:26870731

  7. Virtual Nuclear Envelope Breakdown and Its Regulators in Fission Yeast Meiosis.

    PubMed

    Asakawa, Haruhiko; Yang, Hui-Ju; Hiraoka, Yasushi; Haraguchi, Tokuko

    2016-01-01

    Ran, a small GTPase, is required for the spindle formation and nuclear envelope (NE) formation. After NE breakdown (NEBD) during mitosis in metazoan cells, the Ran-GTP gradient across the NE is lost and Ran-GTP becomes concentrated around chromatin, thus affecting the stability of microtubules and promoting the assembly of spindle microtubules and segregation of chromosomes. Mitosis in which chromosomes are segregated subsequent to NEBD is called "open mitosis." In contrast, many fungi undergo a process termed "closed mitosis" in which chromosome segregation and spindle formation occur without NEBD. Although the fission yeast Schizosaccharomyces pombe undergoes a closed mitosis, it exhibits a short period during meiosis (anaphase of the second meiosis; called "anaphase II") when nuclear and cytoplasmic proteins are mixed in the presence of intact NE and nuclear pore complexes (NPC). This "virtual" nuclear envelope breakdown (vNEBD) involves changes in the localization of RanGAP1, an activator of Ran-GTP hydrolysis. Recently, Nup132, a component of the structural core Nup107-160 subcomplex of the NPC, has been shown to be involved in the maintenance of the nuclear cytoplasmic barrier in yeast meiosis. In this review, we highlight the possible roles of RanGAP1 and Nup132 in vNEBD and discuss the biological significance of vNEBD in S. pombe meiosis.

  8. Role of delayed nuclear envelope breakdown and mitosis in Wolbachia-induced cytoplasmic incompatibility.

    PubMed

    Tram, Uyen; Sullivan, William

    2002-05-10

    The bacterium Wolbachia manipulates reproduction in millions of insects worldwide; the most common effect is cytoplasmic incompatibility (CI). We found that CI resulted from delayed nuclear envelope breakdown of the male pronucleus in Nasonia vitripennis. This caused asynchrony between the male and female pronuclei and, ultimately, loss of paternal chromosomes at the first mitosis. When Wolbachia were present in the egg, synchrony was restored, which explains suppression of CI in these crosses. These results suggest that Wolbachia target cell cycle regulatory proteins. A striking consequence of CI is that it alters the normal pattern of reciprocal centrosome inheritance in Nasonia.

  9. SUN proteins facilitate the removal of membranes from chromatin during nuclear envelope breakdown

    PubMed Central

    Turgay, Yagmur; Champion, Lysie; Balazs, Csaba; Held, Michael; Toso, Alberto; Gerlich, Daniel W.; Meraldi, Patrick

    2014-01-01

    SUN proteins reside in the inner nuclear membrane and form complexes with KASH proteins of the outer nuclear membrane that connect the nuclear envelope (NE) to the cytoskeleton. These complexes have well-established functions in nuclear anchorage and migration in interphase, but little is known about their involvement in mitotic processes. Our analysis demonstrates that simultaneous depletion of human SUN1 and SUN2 delayed removal of membranes from chromatin during NE breakdown (NEBD) and impaired the formation of prophase NE invaginations (PNEIs), similar to microtubule depolymerization or down-regulation of the dynein cofactors NudE/EL. In addition, overexpression of dominant-negative SUN and KASH constructs reduced the occurrence of PNEI, indicating a requirement for functional SUN–KASH complexes in NE remodeling. Codepletion of SUN1/2 slowed cell proliferation and resulted in an accumulation of morphologically defective and disoriented mitotic spindles. Quantification of mitotic timing revealed a delay between NEBD and chromatin separation, indicating a role of SUN proteins in bipolar spindle assembly and mitotic progression. PMID:24662567

  10. Multifunctional Ca2+/calmodulin-dependent protein kinase is necessary for nuclear envelope breakdown

    PubMed Central

    1990-01-01

    The role of multifunctional Ca2+/calmodulin-dependent protein kinase (CaM kinase) in nuclear envelope breakdown (NEB) was investigated in sea urchin eggs. The eggs contain a 56-kD polypeptide which appears to be a homologue of neuronal CaM kinase. For example, it undergoes Ca2+/calmodulin-dependent autophosphorylation that converts it to a Ca2(+)-independent species, a hallmark of multifunctional CaM kinase. It is homologous to the alpha subunit of rat brain CaM kinase. Autophosphorylation and substrate phosphorylation by the sea urchin egg kinase are inhibited in vitro by CaMK(273-302), a synthetic peptide corresponding to the autoinhibitory domain of the neuronal CaM kinase. This peptide inhibited NEB when microinjected into sea urchin eggs. Only one mAb to the neuronal enzyme immunoprecipitated the 56-kD polypeptide. Only this antibody blocked or significantly delayed NEB when microinjected into sea urchin eggs. These results suggest that sea urchin eggs contain multifunctional CaM kinase, and that this enzyme is involved in the control of NEB during mitotic division. PMID:2229172

  11. Meiosis, egg activation, and nuclear envelope breakdown are differentially reliant on Ca2+, whereas germinal vesicle breakdown is Ca2+ independent in the mouse oocyte

    NASA Technical Reports Server (NTRS)

    Tombes, R. M.; Simerly, C.; Borisy, G. G.; Schatten, G.

    1992-01-01

    During early development, intracellular Ca2+ mobilization is not only essential for fertilization, but has also been implicated during other meiotic and mitotic events, such as germinal vesicle breakdown (GVBD) and nuclear envelope breakdown (NEBD). In this study, the roles of intracellular and extracellular Ca2+ were examined during meiotic maturation and reinitiation at parthenogenetic activation and during first mitosis in a single species using the same methodologies. Cumulus-free metaphase II mouse oocytes immediately resumed anaphase upon the induction of a large, transient Ca2+ elevation. This resumption of meiosis and associated events, such as cortical granule discharge, were not sensitive to extracellular Ca2+ removal, but were blocked by intracellular Ca2+ chelators. In contrast, meiosis I was dependent on external Ca2+; in its absence, the formation and function of the first meiotic spindle was delayed, the first polar body did not form and an interphase-like state was induced. GVBD was not dependent on external Ca2+ and showed no associated Ca2+ changes. NEBD at first mitosis in fertilized eggs, on the other hand, was frequently, but not always associated with a brief Ca2+ transient and was dependent on Ca2+ mobilization. We conclude that GVBD is Ca2+ independent, but that the dependence of NEBD on Ca2+ suggests regulation by more than one pathway. As cells develop from Ca(2+)-independent germinal vesicle oocytes to internal Ca(2+)-dependent pronuclear eggs, internal Ca2+ pools increase by approximately fourfold.

  12. Nuclear envelope breakdown may deliver an inhibitor of protein phosphatase 1 which triggers cyclin B translation in starfish oocytes.

    PubMed

    Lapasset, Laure; Pradet-Balade, Bérengère; Lozano, Jean-Claude; Peaucellier, Gérard; Picard, André

    2005-09-01

    In vertebrates, enhanced translation of mRNAs in oocytes and early embryos entering M-phase is thought to occur through polyadenylation, involving binding, hyperphosphorylation and proteolytic degradation of Aurora-activated CPEB. In starfish, an unknown component of the oocyte nucleus is required for cyclin B synthesis following the release of G2/prophase block by hormonal stimulation. We have found that CPEB cannot be hyperphosphorylated following hormonal stimulation in starfish oocytes from which the nucleus has been removed. Activation of Aurora kinase, known to interact with protein phosphatase 1 and its specific inhibitor Inh-2, is also prevented. The microinjection of Inh-2 restores Aurora activation, CPEB hyperphosphorylation and cyclin B translation in enucleated oocytes. Nevertheless, we provide evidence that CPEB is in fact hyperphosphorylated by cdc2, without apparent involvement of Aurora or MAP kinase, and that cyclin B synthesis can be stimulated without previous degradation of phosphorylated CPEB. Thus, the regulation of cyclin B synthesis necessary for progression through meiosis can be explained by an equilibrium between CPEB phosphorylation and dephosphorylation, and both aspects of this control may rely on the sole activation of Cdc2 and subsequent nuclear breakdown.

  13. Targeting Nuclear Envelope Repair.

    PubMed

    2016-06-01

    Migrating cancer cells undergo repeated rupture of the protective nuclear envelope as they squeeze through small spaces in the surrounding tissue, compromising genomic integrity. Inhibiting both general DNA repair and the mechanism that seals these tears may enhance cell death and curb metastasis. PMID:27130435

  14. Cell cycle regulation of human immunodeficiency virus type 1 integration in T cells: antagonistic effects of nuclear envelope breakdown and chromatin condensation

    SciTech Connect

    Mannioui, Abdelkrim . E-mail: karim.mannioui@chu-stlouis.fr; Schiffer, Cecile . E-mail: cecile.schiffer@voila.fr; Felix, Nathalie . E-mail: nathalie.felix@chu-stlouis.fr

    2004-11-10

    We examined the influence of mitosis on the kinetics of human immunodeficiency virus type 1 integration in T cells. Single-round infection of cells arrested in G1b or allowed to synchronously proceed through division showed that mitosis delays virus integration until 18-24 h postinfection, whereas integration reaches maximum levels by 15 h in G1b-arrested cells. Subcellular fractionation of metaphase-arrested cells indicated that, while nuclear envelope disassembly facilitates docking of viral DNA to chromatin, chromosome condensation directly antagonizes and therefore delays integration. As a result of the balance between the two effects, virus integration efficiency is eventually up to threefold greater in dividing cells. At the single-cell level, using a green fluorescent protein-expressing reporter virus, we found that passage through mitosis leads to prominent asymmetric segregation of the viral genome in daughter cells without interfering with provirus expression.

  15. The Arabidopsis Nuclear Pore and Nuclear Envelope

    PubMed Central

    Meier, Iris; Brkljacic, Jelena

    2010-01-01

    The nuclear envelope is a double membrane structure that separates the eukaryotic cytoplasm from the nucleoplasm. The nuclear pores embedded in the nuclear envelope are the sole gateways for macromolecular trafficking in and out of the nucleus. The nuclear pore complexes assembled at the nuclear pores are large protein conglomerates composed of multiple units of about 30 different nucleoporins. Proteins and RNAs traffic through the nuclear pore complexes, enabled by the interacting activities of nuclear transport receptors, nucleoporins, and elements of the Ran GTPase cycle. In addition to directional and possibly selective protein and RNA nuclear import and export, the nuclear pore gains increasing prominence as a spatial organizer of cellular processes, such as sumoylation and desumoylation. Individual nucleoporins and whole nuclear pore subcomplexes traffic to specific mitotic locations and have mitotic functions, for example at the kinetochores, in spindle assembly, and in conjunction with the checkpoints. Mutants of nucleoporin genes and genes of nuclear transport components lead to a wide array of defects from human diseases to compromised plant defense responses. The nuclear envelope acts as a repository of calcium, and its inner membrane is populated by functionally unique proteins connected to both chromatin and—through the nuclear envelope lumen—the cytoplasmic cytoskeleton. Plant nuclear pore and nuclear envelope research—predominantly focusing on Arabidopsis as a model—is discovering both similarities and surprisingly unique aspects compared to the more mature model systems. This chapter gives an overview of our current knowledge in the field and of exciting areas awaiting further exploration. PMID:22303264

  16. The nuclear envelope as a chromatin organizer

    PubMed Central

    Zuleger, Nikolaj; Robson, Michael I

    2011-01-01

    In the past 15 years our perception of nuclear envelope function has evolved perhaps nearly as much as the nuclear envelope itself evolved in the last 3 billion years. Historically viewed as little more than a diffusion barrier between the cytoplasm and the nucleoplasm, the nuclear envelope is now known to have roles in the cell cycle, cytoskeletal stability and cell migration, genome architecture, epigenetics, regulation of transcription, splicing and DNA replication. Here we will review both what is known and what is speculated about the role of the nuclear envelope in genome organization, particularly with respect to the positioning and repositioning of genes and chromosomes within the nucleus during differentiation. PMID:21970986

  17. [NESPRINS--nuclear envelope proteins ensuring integrity].

    PubMed

    Pershina, E G; Morozova, K N; Kiseleva, E V

    2014-01-01

    This review describes the nesprins (nuclear envelope spectrin-repeat proteins), which are recently discovered family of nuclear envelope proteins. These proteins play an important role in maintaining the cellular architecture and establish the link between the nucleus and other sub-cellular compartments. Many tissue-specific diseases including lipodystrophies, hearing loss, cardiac and skeletal myopathies are associated with nesprins mutations. These proteins comprise of multiple tissue specific isoforms which contain spectrin repeats providing interaction of nesprins with other nuclear membrane proteins, cytoskeleton and intranuclear matrix. We summarize recent findings and suggestions about nesprins structural organization and function inside the cell. Human diseases caused by abnormal nesprins expression are also described.

  18. LINCing complex functions at the nuclear envelope

    PubMed Central

    Rothballer, Andrea; Schwartz, Thomas U.; Kutay, Ulrike

    2013-01-01

    Linker of nucleoskeleton and cytoskeleton (LINC) complexes span the double membrane of the nuclear envelope (NE) and physically connect nuclear structures to cytoskeletal elements. LINC complexes are envisioned as force transducers in the NE, which facilitate processes like nuclear anchorage and migration, or chromosome movements. The complexes are built from members of two evolutionary conserved families of transmembrane (TM) proteins, the SUN (Sad1/UNC-84) domain proteins in the inner nuclear membrane (INM) and the KASH (Klarsicht/ANC-1/SYNE homology) domain proteins in the outer nuclear membrane (ONM). In the lumen of the NE, the SUN and KASH domains engage in an intimate assembly to jointly form a NE bridge. Detailed insights into the molecular architecture and atomic structure of LINC complexes have recently revealed the molecular basis of nucleo-cytoskeletal coupling. They bear important implications for LINC complex function and suggest new potential and as yet unexplored roles, which the complexes may play in the cell. PMID:23324460

  19. Regulatory roles of the nuclear envelope.

    PubMed

    Laskey, R A; Görlich, D; Madine, M A; Makkerh, J P; Romanowski, P

    1996-12-15

    Roles of the nuclear envelope are considered in the regulation of nuclear protein import, ribonucleoprotein export, and coupling of DNA replication to the cell cycle. First, evidence is discussed that indicates that neutral and acidic amino acids can be important in nuclear localization signals as well as the widely acknowledged basic amino acids. Second, the recognition of nuclear localization signals by their receptor "importin" is discussed, focusing on the different roles of the two subunits of importin. Third, a role for the alpha subunit of importin in RNP export is considered together with the question of how the direction of traffic through nuclear pores is determined. The final part of this article considers evidence that the nuclear membrane prevents reinitiation of DNA replication in Xenopus eggs, by excluding a "licensing factor" that is essential for DNA replication. Replication licensing in Xenopus appears to involve several proteins including the MCM (minichromosome maintenance) complex and ORC, the origin recognition complex, which must bind before the MCM complex can bind to chromatin. PMID:8986599

  20. Tissue specificity in the nuclear envelope supports its functional complexity

    PubMed Central

    de las Heras, Jose I; Meinke, Peter; Batrakou, Dzmitry G; Srsen, Vlastimil; Zuleger, Nikolaj; Kerr, Alastair RW; Schirmer, Eric C

    2013-01-01

    Nuclear envelope links to inherited disease gave the conundrum of how mutations in near-ubiquitous proteins can yield many distinct pathologies, each focused in different tissues. One conundrum-resolving hypothesis is that tissue-specific partner proteins mediate these pathologies. Such partner proteins may have now been identified with recent proteome studies determining nuclear envelope composition in different tissues. These studies revealed that the majority of the total nuclear envelope proteins are tissue restricted in their expression. Moreover, functions have been found for a number these tissue-restricted nuclear envelope proteins that fit with mechanisms proposed to explain how the nuclear envelope could mediate disease, including defects in mechanical stability, cell cycle regulation, signaling, genome organization, gene expression, nucleocytoplasmic transport, and differentiation. The wide range of functions to which these proteins contribute is consistent with not only their involvement in tissue-specific nuclear envelope disease pathologies, but also tissue evolution. PMID:24213376

  1. Variability of mammalian liver nuclear-envelope preparations.

    PubMed

    Agutter, P S; Gleed, C D

    1980-10-15

    The composition, density and enzymic activities of sheep liver nuclear-envelope preparations were found to vary markedly according to the concentrations of nuclei during the lysis stage. The effect of nuclear concentration on the properties of the purified envelopes could not be attributed to bound Mg2+ or to other ions, and appeared to result from some component of the nucleus which was not eluted during lysis. The implications of these findings for studies on the nuclear envelope are discussed.

  2. Nuclear Envelopes Properties and Physical Interactions with Nucleoplasm

    NASA Astrophysics Data System (ADS)

    Discher, Dennis; Dahl, Kris; Wilson, Kathy

    2004-03-01

    Given the stresses imposed on a cell and its organelles and the nuclear envelope's important role as a barrier between cytoplasm and nucleoplasm, we sought to measure and model mechanical properties of isolated nuclear envelopes. Xenopus laevis oocyte (XO) nuclei are primarily used since they have been widely studied in many fields as model systems for nuclear structure and function. We manipulate the nuclear envelope by both osmotic swelling and micromanipulation to determine an effective elastic modulus. We show the envelope properties are independent of the effects of the nucleoplasm. Micropipette aspiration of XO nuclei gives an effective elastic modulus of the nuclear envelope of 250 mN/m with similar results obtained from isotropic swelling of XO nuclear envelopes. The results suggest that these nuclear envelopes have relatively homogeneous properties and are highly elastic, sustaining strains of 50-100Square-net simulations and comparisons to polymer network models suggests that XO nuclear envelope physical properties are dominated by the lamin network. If applicable to nuclei in other cells, a "pre-compressed" state envisioned here would allow for significant shear flexibility, especially important for motile cells whose nuclei need to rapidly deform.

  3. Variability of mammalian liver nuclear-envelope preparations.

    PubMed Central

    Agutter, P S; Gleed, C D

    1980-01-01

    The composition, density and enzymic activities of sheep liver nuclear-envelope preparations were found to vary markedly according to the concentrations of nuclei during the lysis stage. The effect of nuclear concentration on the properties of the purified envelopes could not be attributed to bound Mg2+ or to other ions, and appeared to result from some component of the nucleus which was not eluted during lysis. The implications of these findings for studies on the nuclear envelope are discussed. Images Fig. 3. PMID:7305915

  4. The nuclear envelope proteome differs notably between tissues

    PubMed Central

    Korfali, Nadia; Wilkie, Gavin S.; Swanson, Selene K.; Srsen, Vlastimil; de las Heras, Jose; Batrakou, Dzmitry G.; Malik, Poonam; Zuleger, Nikolaj; Kerr, Alastair R.W.; Florens, Laurence; Schirmer, Eric C.

    2012-01-01

    One hypothesis to explain how mutations in the same nuclear envelope proteins yield pathologies focused in distinct tissues is that as yet unidentified tissue-specific partners mediate the disease pathologies. The nuclear envelope proteome was recently determined from leukocytes and muscle. Here the same methodology is applied to liver and a direct comparison of the liver, muscle and leukocyte data sets is presented. At least 74 novel transmembrane proteins identified in these studies have been directly confirmed at the nuclear envelope. Within this set, RT-PCR, western blot and staining of tissue cryosections confirms that the protein complement of the nuclear envelope is clearly distinct from one tissue to another. Bioinformatics reveals similar divergence between tissues across the larger data sets. For proteins acting in complexes according to interactome data, the whole complex often exhibited the same tissue-specificity. Other tissue-specific nuclear envelope proteins identified were known proteins with functions in signaling and gene regulation. The high tissue specificity in the nuclear envelope likely underlies the complex disease pathologies and argues that all organelle proteomes warrant re-examination in multiple tissues. PMID:22990521

  5. Exploring the Protein Composition of the Plant Nuclear Envelope.

    PubMed

    Zhou, Xiao; Tamura, Kentaro; Graumann, Katja; Meier, Iris

    2016-01-01

    Due to rather limited sequence similarity, targeted identification of plant nuclear envelope and nuclear pore complex proteins has mainly followed two routes: (1) advanced computational identification followed by experimental verification and (2) immunoaffinity purification of complexes followed by mass spectrometry. Following candidate identification, fluorescence recovery after photobleaching (FRAP) and fluorescence resonance energy transfer (FRET) provide powerful tools to verify protein-protein interactions in situ at the NE. Here, we describe these methods for the example of Arabidopsis thaliana nuclear pore and nuclear envelope protein identification.

  6. Jumping the nuclear envelop barrier: Improving polyplex-mediated gene transfection efficiency by a selective CDK1 inhibitor RO-3306.

    PubMed

    Zhou, Xuefei; Liu, Xiangrui; Zhao, Bingxiang; Liu, Xin; Zhu, Dingcheng; Qiu, Nasha; Zhou, Quan; Piao, Ying; Zhou, Zhuxian; Tang, Jianbin; Shen, Youqing

    2016-07-28

    Successful transfection of plasmid DNA (pDNA) requires intranuclear internalization of pDNA effectively and the nuclear envelope appears to be one of the critical intracellular barriers for polymer mediated pDNA delivery. Polyethylenimine (PEI), as the classic cationic polymer, compact the negatively charged pDNA tightly and make up stable polyplexes. The polyplexes are too large to enter the nuclear through nuclear pores and it is believed that the nuclear envelope breakdown in mitosis could facilitate the nuclear entry of polyplexes. To jump the nuclear envelope barrier, we used a selective and reversible CDK1 inhibitor RO-3306 to control the G2/M transition of the cell cycle and increased the proportion of mitotic cells which have disappeared nuclear envelope during transfection. Herein, we show that RO-3306 remarkably increases the transfection efficiency of PEI polyplexes through enhanced nuclear localization of PEI and pDNA. However, RO-3306 is less effective to the charge-reversal polymer poly[(2-acryloyl)ethyl(p-boronic acid benzyl)diethylammonium bromide] (B-PDEAEA) which responses to cellular stimuli and releases free pDNA in cytoplasm. Our findings not only offer new opportunities for improving non-viral based gene delivery but also provide theoretical support for the rational design of novel functional polymers for gene delivery. We also report current data showing that RO-3306 synergizes TRAIL gene induced apoptosis in cancer cells.

  7. Efflux of RNA from resealed nuclear envelope ghosts.

    PubMed

    Prochnow, D; Thomson, M; Schröder, H C; Müller, W E; Agutter, P S

    1994-08-01

    mRNA translocation across the nuclear envelope and the appropriate signal-receptor interactions have been studied using resealed rat liver nuclear envelope ghosts (RNEG). We compared export kinetics of nonadenylated (tRNAs, histone-2 poly(A)- mRNA), and adenylated RNAs (poly(A)+ tRNAs, synthetic histone-2 poly(A) +mRNA, albumin mRNA, beta-globin poly(A) +mRNA and a total poly(A) + mRNA extract from rat liver cells). ATP-dependent export of mRNAs and of total poly(A)+ RNA was prevented by inhibitors of a nuclear envelope NTPase. All adenylated RNA species competed with each other for export, but nonadenylated RNAs did not. This indicates the existence of different translocation mechanisms for different RNA species with their appropriate nuclear envelope associated RNA receptors involved in export. The attachment of a poly(A)250 sequence at the 3'-end of tRNA or histone messenger masks the intrinsic RNA export signal of nonadenylated RNAs and results in efflux comparable to that of beta-globin poly(A)+ mRNA. The attachment on oligo(A)5 does not have any comparable effect of nonadenylated RNA translocation. Export of all polyadenylated RNAs from RNEGs is blocked by a monoclonal antibody, which is directed against an intranuclear envelope poly(A) binding protein. The results suggest that the pore complexes do not select RNAs for export to the cytoplasm and are therefore not responsible for nuclear restriction of mRNA precursors.

  8. The nuclear envelope lamina is reversibly depolymerized during mitosis.

    PubMed

    Gerace, L; Blobel, G

    1980-01-01

    The nuclear envelope lamina is a supramolecular protein assembly associated with the nucleoplasmic surface of the inner nuclear membrane, which contains three predominant polypeptide components in mammalian cells (lamins A, B and C). We previously demonstrated by immunofluorescence microscopy that the lamina is reversibly disassembled during cell division, coincident with the disassembly and reconstruction of the mitotic nuclear envelope architecture. In this paper, these immunocytochemical observations are extended with cell fractionation and immunoprecipitation studies performed on synchronized populations of tissue culture cells. With these techniques, we have established that during mitosis, lamina A and C occur in a soluble and nonmembrane-associated state. In contrast, the mitotic lamin B may be associated with membrane fragments derived from the disassembled interphase nuclear envelope. From sedimentation analysis on sucrose gradients, we have determined that all three lamins are monomeric at periods of mitotic lamina disassembly. These results, together with quantitative immunoprecipitation studies, demonstrate that the lamina is reversibly depolymerized during cell division. Attendant with the depolymerized state of the lamina, the mitotic lamins (which are phosphoproteins) have a distinctly more acidic isoelectric point and a substantially higher level of phosphorylation compared to their interphase counterparts. This indicates that reversible enzymatic phosphorylations of the lamins may be involved in modulating the state of polymerization of the lamina and its reversible mitotic disassembly. PMID:7357605

  9. Fluctuations in nuclear envelope's potential mediate synchronization of early neural activity

    SciTech Connect

    Yamashita, Masayuki

    2011-03-04

    Research highlights: {yields} Nuclear envelope's potential changes with a release of Ca{sup 2+}. {yields} Changes in nuclear envelope's potential underlie synchronous burst discharges. {yields} Nuclear envelope's potential generates periodic bursts of fluctuations. {yields} Fluctuations in nuclear envelope's potential function as a current noise generator. -- Abstract: Neural progenitor cells and developing neurons show periodic, synchronous Ca{sup 2+} rises even before synapse formation, and the origin of the synchronous activity remains unknown. Here, fluorescence measurement revealed that the membrane potential of the nuclear envelope, which forms an intracellular Ca{sup 2+} store, changed with a release of Ca{sup 2+} and generated spontaneous, periodic bursts of fluctuations in potential. Furthermore, changes in the nuclear envelope's potential underlay spike burst generations. These results support the model that voltage fluctuations of the nuclear envelope synchronize Ca{sup 2+} release between cells and also function as a current noise generator to cause synchronous burst discharges.

  10. Ribonucleic acid stimulation of mammalian liver nuclear-envelope nucleoside triphosphatase. A possible enzymic marker for the nuclear envelope.

    PubMed Central

    Agutter, P S; Harris, J R; Stevenson, I

    1977-01-01

    1. The specific activity of rat and pig liver nuclear-envelope nucleoside triphosphatase (EC 3.6.1.3) decreases when the system is depleted of RNA. The activity can be restored by adding high concentrations of yeast RNA to the assay medium. 2. Exogenous RNA also increases the activity of the enzyme in control envelopes (not RNA-depleted). The effect appears to be largely specific for poly(A) and poly(G); it is not stimulated by rRNA or tRNA preparations, ribonuclease-hydrolysed RNA, AMP, or double- or single-stranded DNA. 3. Inhibitors of the enzyme, in concentrations at which half-maximal inhibition of the enzyme is achieved, do not affect the percentage stimulation of the enzyme by yeast RNA. 4. The simulation is abolished by the inclusion of 150 mM-KCl or -NaCl in the assay medium, but not by increasing the assay pH to 8.5. 5. The results are discussed in the light of the possible role of the nucleoside triphosphatase in vivo in nucleo-cytoplasmic ribonucleoprotein translocation. 6. It is proposed that poly(G)-stimulated Mg2+-activated adenosine triphosphatase activity should be adopted as an enzymic marker for the nuclear envelope. Images PLATE 1 PMID:141276

  11. Ribonucleic acid stimulation of mammalian liver nuclear-envelope nucleoside triphosphatase. A possible enzymic marker for the nuclear envelope.

    PubMed

    Agutter, P S; Harris, J R; Stevenson, I

    1977-03-15

    1. The specific activity of rat and pig liver nuclear-envelope nucleoside triphosphatase (EC 3.6.1.3) decreases when the system is depleted of RNA. The activity can be restored by adding high concentrations of yeast RNA to the assay medium. 2. Exogenous RNA also increases the activity of the enzyme in control envelopes (not RNA-depleted). The effect appears to be largely specific for poly(A) and poly(G); it is not stimulated by rRNA or tRNA preparations, ribonuclease-hydrolysed RNA, AMP, or double- or single-stranded DNA. 3. Inhibitors of the enzyme, in concentrations at which half-maximal inhibition of the enzyme is achieved, do not affect the percentage stimulation of the enzyme by yeast RNA. 4. The simulation is abolished by the inclusion of 150 mM-KCl or -NaCl in the assay medium, but not by increasing the assay pH to 8.5. 5. The results are discussed in the light of the possible role of the nucleoside triphosphatase in vivo in nucleo-cytoplasmic ribonucleoprotein translocation. 6. It is proposed that poly(G)-stimulated Mg2+-activated adenosine triphosphatase activity should be adopted as an enzymic marker for the nuclear envelope.

  12. Nuclear envelope rupture and repair during cancer cell migration

    PubMed Central

    Denais, Celine M.; Gilbert, Rachel M.; Isermann, Philipp; McGregor, Alexandra L.; te Lindert, Mariska; Weigelin, Bettina; Davidson, Patricia M.; Friedl, Peter; Wolf, Katarina; Lammerding, Jan

    2016-01-01

    During cancer metastasis, tumor cells penetrate tissues through tight interstitial spaces, requiring extensive deformation of the cell and its nucleus. Here, we investigated tumor cell migration in confining microenvironments in vitro and in vivo. Nuclear deformation caused localized loss of nuclear envelope (NE) integrity, which led to the uncontrolled exchange of nucleo-cytoplasmic content, herniation of chromatin across the NE, and DNA damage. The incidence of NE rupture increased with cell confinement and with depletion of nuclear lamins, NE proteins that structurally support the nucleus. Cells restored NE integrity using components of the endosomal sorting complexes required for transport-III (ESCRT-III) machinery. Our findings indicate that cell migration incurs substantial physical stress on the NE and its content, requiring efficient NE and DNA damage repair for survival. PMID:27013428

  13. LINC'ing form and function at the nuclear envelope.

    PubMed

    Meinke, Peter; Schirmer, Eric C

    2015-09-14

    The nuclear envelope is an amazing piece of engineering. On one hand it is built like a mediaeval fortress with filament systems reinforcing its membrane walls and its double membrane structure forming a lumen like a castle moat. On the other hand its structure can adapt while maintaining its integrity like a reed bending in a river. Like a fortress it has guarded drawbridges in the nuclear pore complexes, but also has other mechanical means of communication. All this is enabled largely because of the LINC complex, a multi-protein structure that connects the intermediate filament nucleoskeleton across the lumen of the double membrane nuclear envelope to multiple cytoplasmic filament systems that themselves could act simultaneously both like mediaeval buttresses and like lines on a suspension bridge. Although many details of the greater LINC structure remain to be discerned, a number of recent findings are giving clues as to how its structural organization can yield such striking dynamic yet stable properties. Combining double- and triple-helical coiled-coils, intrinsic disorder and order, tissue-specific components, and intermediate filaments enables these unique properties. PMID:26096784

  14. Several Novel Nuclear Envelope Transmembrane Proteins Identified in Skeletal Muscle Have Cytoskeletal Associations*

    PubMed Central

    Wilkie, Gavin S.; Korfali, Nadia; Swanson, Selene K.; Malik, Poonam; Srsen, Vlastimil; Batrakou, Dzmitry G.; de las Heras, Jose; Zuleger, Nikolaj; Kerr, Alastair R. W.; Florens, Laurence; Schirmer, Eric C.

    2011-01-01

    Nuclear envelopes from liver and a neuroblastoma cell line have previously been analyzed by proteomics; however, most diseases associated with the nuclear envelope affect muscle. To determine whether muscle has unique nuclear envelope proteins, rat skeletal muscle nuclear envelopes were prepared and analyzed by multidimensional protein identification technology. Many novel muscle-specific proteins were identified that did not appear in previous nuclear envelope data sets. Nuclear envelope residence was confirmed for 11 of these by expression of fusion proteins and by antibody staining of muscle tissue cryosections. Moreover, transcript levels for several of the newly identified nuclear envelope transmembrane proteins increased during muscle differentiation using mouse and human in vitro model systems. Some of these proteins tracked with microtubules at the nuclear surface in interphase cells and accumulated at the base of the microtubule spindle in mitotic cells, suggesting they may associate with complexes that connect the nucleus to the cytoskeleton. The finding of tissue-specific proteins in the skeletal muscle nuclear envelope proteome argues the importance of analyzing nuclear envelopes from all tissues linked to disease and suggests that general investigation of tissue differences in organellar proteomes might yield critical insights. PMID:20876400

  15. Several novel nuclear envelope transmembrane proteins identified in skeletal muscle have cytoskeletal associations.

    PubMed

    Wilkie, Gavin S; Korfali, Nadia; Swanson, Selene K; Malik, Poonam; Srsen, Vlastimil; Batrakou, Dzmitry G; de las Heras, Jose; Zuleger, Nikolaj; Kerr, Alastair R W; Florens, Laurence; Schirmer, Eric C

    2011-01-01

    Nuclear envelopes from liver and a neuroblastoma cell line have previously been analyzed by proteomics; however, most diseases associated with the nuclear envelope affect muscle. To determine whether muscle has unique nuclear envelope proteins, rat skeletal muscle nuclear envelopes were prepared and analyzed by multidimensional protein identification technology. Many novel muscle-specific proteins were identified that did not appear in previous nuclear envelope data sets. Nuclear envelope residence was confirmed for 11 of these by expression of fusion proteins and by antibody staining of muscle tissue cryosections. Moreover, transcript levels for several of the newly identified nuclear envelope transmembrane proteins increased during muscle differentiation using mouse and human in vitro model systems. Some of these proteins tracked with microtubules at the nuclear surface in interphase cells and accumulated at the base of the microtubule spindle in mitotic cells, suggesting they may associate with complexes that connect the nucleus to the cytoskeleton. The finding of tissue-specific proteins in the skeletal muscle nuclear envelope proteome argues the importance of analyzing nuclear envelopes from all tissues linked to disease and suggests that general investigation of tissue differences in organellar proteomes might yield critical insights. PMID:20876400

  16. Properties of mammalian nuclear-envelope nucleoside triphosphatase.

    PubMed Central

    Agutter, P S; Cockrill, J B; Lavine, J E; McCaldin, B; Sim, R B

    1979-01-01

    The nucleoside triphosphatase activities of the nuclear envelopes from rat liver, pig liver and simian-virus-40-transformed mouse-embryo 3T3 cells were shown to exhibit similar parperties. All three preparations hydrolyse ATP, 2'-dATP, 3'-dATP, GTP, CTP and UTP in the presence of Mg2+, Ca2+, Mn2+ and Co2+ with a pH optimum of 8.0, are sensitive to inhibition by mercurials, arsenicals, quercetin, proflavin and adenosine 5'-[gamma-thio]triphosphate and are partially inactivated by exposure to high ionic strength. The kinetic behaviour is similar for all substrates irrespective of the source of material. The typical Eadie-Hofstee plot, which is concave upwards at pH 8.0 when the ionic strength is 20mM, becomes linear when the pH is increased to 8.5 or the ionic strength to 160mM. The overall evidence, particularly the labelling of only one polypeptide by [gamma-32P]ATP, suggests that under the conditions of preparation and assay used only one class of nucleoside triphosphatase active sites is detectable in nuclear envelopes. The importance of these results for an understanding of the role of the enzyme in vivo is discussed. PMID:229821

  17. Properties of mammalian nuclear-envelope nucleoside triphosphatase.

    PubMed

    Agutter, P S; Cockrill, J B; Lavine, J E; McCaldin, B; Sim, R B

    1979-09-01

    The nucleoside triphosphatase activities of the nuclear envelopes from rat liver, pig liver and simian-virus-40-transformed mouse-embryo 3T3 cells were shown to exhibit similar parperties. All three preparations hydrolyse ATP, 2'-dATP, 3'-dATP, GTP, CTP and UTP in the presence of Mg2+, Ca2+, Mn2+ and Co2+ with a pH optimum of 8.0, are sensitive to inhibition by mercurials, arsenicals, quercetin, proflavin and adenosine 5'-[gamma-thio]triphosphate and are partially inactivated by exposure to high ionic strength. The kinetic behaviour is similar for all substrates irrespective of the source of material. The typical Eadie-Hofstee plot, which is concave upwards at pH 8.0 when the ionic strength is 20mM, becomes linear when the pH is increased to 8.5 or the ionic strength to 160mM. The overall evidence, particularly the labelling of only one polypeptide by [gamma-32P]ATP, suggests that under the conditions of preparation and assay used only one class of nucleoside triphosphatase active sites is detectable in nuclear envelopes. The importance of these results for an understanding of the role of the enzyme in vivo is discussed.

  18. Cytosol-dependent membrane fusion in ER, nuclear envelope and nuclear pore assembly: biological implications.

    PubMed

    Rafikova, Elvira R; Melikov, Kamran; Chernomordik, Leonid V

    2010-01-01

    Endoplasmic reticulum and nuclear envelope rearrangements after mitosis are often studied in the reconstitution system based on Xenopus egg extract. In our recent work we partially replaced the membrane vesicles in the reconstitution mix with protein-free liposomes to explore the relative contributions of cytosolic and transmembrane proteins. Here we discuss our finding that cytosolic proteins mediate fusion between membranes lacking functional transmembrane proteins and the role of membrane fusion in endoplasmic reticulum and nuclear envelope reorganization. Cytosol-dependent liposome fusion has allowed us to restore, without adding transmembrane nucleoporins, functionality of nuclear pores, their spatial distribution and chromatin decondensation in nuclei formed at insufficient amounts of membrane material and characterized by only partial decondensation of chromatin and lack of nuclear transport. Both the mechanisms and the biological implications of the discovered coupling between spatial distribution of nuclear pores, chromatin decondensation and nuclear transport are discussed.

  19. NET23/STING Promotes Chromatin Compaction from the Nuclear Envelope

    PubMed Central

    de las Heras, Jose I.; Saiz-Ros, Natalia; Makarov, Alexandr A.; Lazou, Vassiliki; Meinke, Peter; Waterfall, Martin; Kelly, David A.; Schirmer, Eric C.

    2014-01-01

    Changes in the peripheral distribution and amount of condensed chromatin are observed in a number of diseases linked to mutations in the lamin A protein of the nuclear envelope. We postulated that lamin A interactions with nuclear envelope transmembrane proteins (NETs) that affect chromatin structure might be altered in these diseases and so screened thirty-one NETs for those that promote chromatin compaction as determined by an increase in the number of chromatin clusters of high pixel intensity. One of these, NET23 (also called STING, MITA, MPYS, ERIS, Tmem173), strongly promoted chromatin compaction. A correlation between chromatin compaction and endogenous levels of NET23/STING was observed for a number of human cell lines, suggesting that NET23/STING may contribute generally to chromatin condensation. NET23/STING has separately been found to be involved in innate immune response signaling. Upon infection cells make a choice to either apoptose or to alter chromatin architecture to support focused expression of interferon genes and other response factors. We postulate that the chromatin compaction induced by NET23/STING may contribute to this choice because the cells expressing NET23/STING eventually apoptose, but the chromatin compaction effect is separate from this as the condensation was still observed when cells were treated with Z-VAD to block apoptosis. NET23/STING-induced compacted chromatin revealed changes in epigenetic marks including changes in histone methylation and acetylation. This indicates a previously uncharacterized nuclear role for NET23/STING potentially in both innate immune signaling and general chromatin architecture. PMID:25386906

  20. NET23/STING promotes chromatin compaction from the nuclear envelope.

    PubMed

    Malik, Poonam; Zuleger, Nikolaj; de las Heras, Jose I; Saiz-Ros, Natalia; Makarov, Alexandr A; Lazou, Vassiliki; Meinke, Peter; Waterfall, Martin; Kelly, David A; Schirmer, Eric C

    2014-01-01

    Changes in the peripheral distribution and amount of condensed chromatin are observed in a number of diseases linked to mutations in the lamin A protein of the nuclear envelope. We postulated that lamin A interactions with nuclear envelope transmembrane proteins (NETs) that affect chromatin structure might be altered in these diseases and so screened thirty-one NETs for those that promote chromatin compaction as determined by an increase in the number of chromatin clusters of high pixel intensity. One of these, NET23 (also called STING, MITA, MPYS, ERIS, Tmem173), strongly promoted chromatin compaction. A correlation between chromatin compaction and endogenous levels of NET23/STING was observed for a number of human cell lines, suggesting that NET23/STING may contribute generally to chromatin condensation. NET23/STING has separately been found to be involved in innate immune response signaling. Upon infection cells make a choice to either apoptose or to alter chromatin architecture to support focused expression of interferon genes and other response factors. We postulate that the chromatin compaction induced by NET23/STING may contribute to this choice because the cells expressing NET23/STING eventually apoptose, but the chromatin compaction effect is separate from this as the condensation was still observed when cells were treated with Z-VAD to block apoptosis. NET23/STING-induced compacted chromatin revealed changes in epigenetic marks including changes in histone methylation and acetylation. This indicates a previously uncharacterized nuclear role for NET23/STING potentially in both innate immune signaling and general chromatin architecture. PMID:25386906

  1. NET23/STING promotes chromatin compaction from the nuclear envelope.

    PubMed

    Malik, Poonam; Zuleger, Nikolaj; de las Heras, Jose I; Saiz-Ros, Natalia; Makarov, Alexandr A; Lazou, Vassiliki; Meinke, Peter; Waterfall, Martin; Kelly, David A; Schirmer, Eric C

    2014-01-01

    Changes in the peripheral distribution and amount of condensed chromatin are observed in a number of diseases linked to mutations in the lamin A protein of the nuclear envelope. We postulated that lamin A interactions with nuclear envelope transmembrane proteins (NETs) that affect chromatin structure might be altered in these diseases and so screened thirty-one NETs for those that promote chromatin compaction as determined by an increase in the number of chromatin clusters of high pixel intensity. One of these, NET23 (also called STING, MITA, MPYS, ERIS, Tmem173), strongly promoted chromatin compaction. A correlation between chromatin compaction and endogenous levels of NET23/STING was observed for a number of human cell lines, suggesting that NET23/STING may contribute generally to chromatin condensation. NET23/STING has separately been found to be involved in innate immune response signaling. Upon infection cells make a choice to either apoptose or to alter chromatin architecture to support focused expression of interferon genes and other response factors. We postulate that the chromatin compaction induced by NET23/STING may contribute to this choice because the cells expressing NET23/STING eventually apoptose, but the chromatin compaction effect is separate from this as the condensation was still observed when cells were treated with Z-VAD to block apoptosis. NET23/STING-induced compacted chromatin revealed changes in epigenetic marks including changes in histone methylation and acetylation. This indicates a previously uncharacterized nuclear role for NET23/STING potentially in both innate immune signaling and general chromatin architecture.

  2. Lipid partitioning at the nuclear envelope controls membrane biogenesis

    PubMed Central

    Barbosa, Antonio Daniel; Sembongi, Hiroshi; Su, Wen-Min; Abreu, Susana; Reggiori, Fulvio; Carman, George M.; Siniossoglou, Symeon

    2015-01-01

    Partitioning of lipid precursors between membranes and storage is crucial for cell growth, and its disruption underlies pathologies such as cancer, obesity, and type 2 diabetes. However, the mechanisms and signals that regulate this process are largely unknown. In yeast, lipid precursors are mainly used for phospholipid synthesis in nutrient-rich conditions in order to sustain rapid proliferation but are redirected to triacylglycerol (TAG) stored in lipid droplets during starvation. Here we investigate how cells reprogram lipid metabolism in the endoplasmic reticulum. We show that the conserved phosphatidate (PA) phosphatase Pah1, which generates diacylglycerol from PA, targets a nuclear membrane subdomain that is in contact with growing lipid droplets and mediates TAG synthesis. We find that cytosol acidification activates the master regulator of Pah1, the Nem1-Spo7 complex, thus linking Pah1 activity to cellular metabolic status. In the absence of TAG storage capacity, Pah1 still binds the nuclear membrane, but lipid precursors are redirected toward phospholipids, resulting in nuclear deformation and a proliferation of endoplasmic reticulum membrane. We propose that, in response to growth signals, activation of Pah1 at the nuclear envelope acts as a switch to control the balance between membrane biogenesis and lipid storage. PMID:26269581

  3. A trypsin-sensitive receptor on membrane vesicles is required for nuclear envelope formation in vitro

    PubMed Central

    1988-01-01

    The reformation of functioning organelles at the end of mitosis presents a problem in vesicle targeting. Using extracts made from Xenopus laevis frog eggs, we have studied in vitro the vesicles that reform the nuclear envelope. In the in vitro assay, nuclear envelope growth is linear with time. Furthermore, the final surface area of the nuclear envelopes formed is directly dependent upon the amount of membrane vesicles added to the assay. Egg membrane vesicles could be fractionated into two populations, only one of which was competent for nuclear envelope assembly. We found that vesicles active in nuclear envelope assembly contained markers (BiP and alpha-glucosidase II) characteristic of the endoplasmic reticulum (ER), but that the majority of ER-derived vesicles do not contribute to nuclear envelope size. This functional distinction between nuclear vesicles and ER-derived vesicles implies that nuclear vesicles are unique and possess at least one factor required for envelope assembly that is lacking in other vesicles. Consistent with this, treatment of vesicles with trypsin destroyed their ability to form a nuclear envelope; electron microscopic studies indicate that the trypsin-sensitive proteins is required for vesicles to bind to chromatin. However, the protease- sensitive component(s) is resistant to treatments that disrupt protein- protein interactions, such as high salt, EDTA, or low ionic strength solutions. We propose that an integral membrane protein, or protein tightly associated with the membrane, is critical for nuclear vesicle targeting or function. PMID:3392106

  4. Nuclear envelope and genome interactions in cell fate

    PubMed Central

    Talamas, Jessica A.; Capelson, Maya

    2015-01-01

    The eukaryotic cell nucleus houses an organism’s genome and is the location within the cell where all signaling induced and development-driven gene expression programs are ultimately specified. The genome is enclosed and separated from the cytoplasm by the nuclear envelope (NE), a double-lipid membrane bilayer, which contains a large variety of trans-membrane and associated protein complexes. In recent years, research regarding multiple aspects of the cell nucleus points to a highly dynamic and coordinated concert of efforts between chromatin and the NE in regulation of gene expression. Details of how this concert is orchestrated and how it directs cell differentiation and disease are coming to light at a rapid pace. Here we review existing and emerging concepts of how interactions between the genome and the NE may contribute to tissue specific gene expression programs to determine cell fate. PMID:25852741

  5. GTP-binding proteins in rat liver nuclear envelopes.

    PubMed Central

    Rubins, J B; Benditt, J O; Dickey, B F; Riedel, N

    1990-01-01

    Nuclear transport as well as reassembly of the nuclear envelope (NE) after completion of mitosis are processes that have been shown to require GTP and ATP. To study the presence and localization of GTP-binding proteins in the NE, we have combined complementary techniques of [alpha-32P]GTP binding to Western-blotted proteins and UV crosslinking of [alpha-32P]GTP with well-established procedures for NE subfractionation. GTP binding to blotted NE proteins revealed five low molecular mass GTP-binding proteins of 26, 25, 24.5, 24, and 23 kDa, and [alpha-32P]GTP photoaffinity labeling revealed major proteins with apparent molecular masses of 140, 53, 47, 33, and 31 kDa. All GTP-binding proteins appear to localize preferentially to the inner nuclear membrane, possibly to the interface between inner nuclear membrane and lamina. Despite the evolutionary conservation between the NE and the rough endoplasmic reticulum, the GTP-binding proteins identified differed between these two compartments. Most notably, the 68- and 30-kDa GTP-binding subunits of the signal recognition particle receptor, which photolabeled with [alpha-32P]GTP in the rough endoplasmic reticulum fraction, were totally excluded from the NE fraction. Conversely, a major 53-kDa photolabeled protein in the NE was absent from rough endoplasmic reticulum. Whereas Western-blotted NE proteins bound GTP specifically, all [alpha-32P]GTP photolabeled proteins could be blocked by competition with ATP, although with a competition profile that differed from that obtained with GTP. In comparative crosslinking studies with [alpha-32P]ATP, we have identified three specific ATP-binding proteins with molecular masses of 160, 78, and 74 kDa. The localization of GTP- and ATP-binding proteins within the NE appears appropriate for their involvement in nuclear transport and in the GTP-dependent fusion of nuclear membrane vesicles required for reassembly of the nucleus after mitosis. Images PMID:2119502

  6. Cytomegalovirus Primary Envelopment Occurs at Large Infoldings of the Inner Nuclear Membrane▿

    PubMed Central

    Buser, Christopher; Walther, Paul; Mertens, Thomas; Michel, Detlef

    2007-01-01

    We have investigated the morphogenesis of human and murine cytomegalovirus by transmission electron microscopy after high-pressure freezing, freeze substitution, and plastic embedding. We observed large tubular infoldings of the inner nuclear membrane that were free of lamina and active in primary envelopment and subsequent transport of capsids to the nuclear periphery. Semiquantitative determinations of the enlarged inner nuclear membrane area and the location of the primary envelopment of nucleocapsids demonstrated that this structure represents a virus-induced specialized membrane domain at which the particles are preferentially enveloped. This is a previously undescribed structural element relevant in cytomegalovirus morphogenesis. PMID:17192309

  7. [Glucose-6-phosphatase from nuclear envelope in rat liver].

    PubMed

    González-Mujica, Freddy

    2008-06-01

    Nuclear envelope (NE) and microsomal glucosa-6-phosphatase (G-6-Pase) activities were compared. Intact microsomes were unable to hydrolyze mannose-6-phosphate (M-6-P), on the other hand, intact NE hydrolyzes this substrate. Galactose-6-phosphate showed to be a good substrate for both NE and microsomal enzymes, with similar latency to that obtained with M-6-P using microsomes. In consequence, this substrate was used to measure the NE integrity. The kinetic parameters (Kii and Kis) of the intact NE G-6-Pase for the phlorizin inhibition using glucose-6-phosphate (G-6-P) and M-6-P as substrates, were very similar. The NE T1 transporter was more sensitive to amiloride than the microsomal T1. The microsomal system was more sensitive to N-ethylmalemide (NEM) than the NE and the latter was insensitive to anion transport inhibitors DIDS and SITS, which strongly affect the microsomal enzyme. The above results allowed to postulate the presence of a hexose-6-phosphate transporter in the NE which is able to carry G-6-P and M-6-P, and perhaps other hexose-6-phosphate which could be different from that present in microsomes or, if it is the same, its activity could by modified by the membrane system where it is included. The higher PPi hydrolysis activity of the intact NE G-6-Pase in comparison to the intact microsomal, suggests differences between the Pi/PPi transport (T2) of both systems. The lower sensitivity of the NE G-6-Pase to NEM suggests that the catalytic subunit of this system has some differences with the microsomal isoform. PMID:18717264

  8. MCLIP Detection of Novel Protein-Protein Interactions at the Nuclear Envelope.

    PubMed

    Jafferali, Mohammed Hakim; Figueroa, Ricardo A; Hallberg, Einar

    2016-01-01

    The organization and function of the nuclear envelope (NE) involves hundreds of nuclear membrane proteins and myriad protein-protein interactions, most of which are still uncharacterized. Many NE proteins interact stably or dynamically with the nuclear lamina or chromosomes. This can make them difficult to extract under nondenaturing conditions, and greatly limits our ability to explore and identify functional protein interactions at the NE. This knowledge is needed to understand nuclear envelope structure and the mechanisms of human laminopathy diseases. This chapter provides detailed protocols for MCLIP (membrane cross-linking immunoprecipitation) identification of novel protein-protein interactions in mammalian cells.

  9. Sizing up the nucleus: nuclear shape, size and nuclear-envelope assembly

    PubMed Central

    Webster, Micah; Witkin, Keren L.; Cohen-Fix, Orna

    2009-01-01

    Summary The nucleus is one of the most prominent cellular organelles, yet surprisingly little is known about how it is formed, what determines its shape and what defines its size. As the nuclear envelope (NE) disassembles in each and every cell cycle in metazoans, the process of rebuilding the nucleus is crucial for proper development and cell proliferation. In this Commentary, we summarize what is known about the regulation of nuclear shape and size, and highlight recent findings that shed light on the process of building a nucleus, including new discoveries related to NE assembly and the relationship between the NE and the endoplasmic reticulum (ER). Throughout our discussion, we note interesting aspects of nuclear structure that have yet to be resolved. Finally, we present an idea – which we refer to as `the limited flat membrane hypothesis' – to explain the formation of a single nucleus that encompasses of all of the cell's chromosomes following mitosis. PMID:19420234

  10. Pre-assembled Nuclear Pores Insert into the Nuclear Envelope during Early Development.

    PubMed

    Hampoelz, Bernhard; Mackmull, Marie-Therese; Machado, Pedro; Ronchi, Paolo; Bui, Khanh Huy; Schieber, Nicole; Santarella-Mellwig, Rachel; Necakov, Aleksandar; Andrés-Pons, Amparo; Philippe, Jean Marc; Lecuit, Thomas; Schwab, Yannick; Beck, Martin

    2016-07-28

    Nuclear pore complexes (NPCs) span the nuclear envelope (NE) and mediate nucleocytoplasmic transport. In metazoan oocytes and early embryos, NPCs reside not only within the NE, but also at some endoplasmic reticulum (ER) membrane sheets, termed annulate lamellae (AL). Although a role for AL as NPC storage pools has been discussed, it remains controversial whether and how they contribute to the NPC density at the NE. Here, we show that AL insert into the NE as the ER feeds rapid nuclear expansion in Drosophila blastoderm embryos. We demonstrate that NPCs within AL resemble pore scaffolds that mature only upon insertion into the NE. We delineate a topological model in which NE openings are critical for AL uptake that nevertheless occurs without compromising the permeability barrier of the NE. We finally show that this unanticipated mode of pore insertion is developmentally regulated and operates prior to gastrulation. PMID:27397507

  11. Isolation, Proteomic Analysis, and Microscopy Confirmation of the Liver Nuclear Envelope Proteome.

    PubMed

    Korfali, Nadia; Florens, Laurence; Schirmer, Eric C

    2016-01-01

    Nuclei can be relatively easily extracted from homogenized liver due to the softness of the tissue and crudely separated from other cellular organelles by low-speed centrifugation due to the comparatively large size of nuclei. However, further purification is complicated by nuclear envelope continuity with the endoplasmic reticulum, invaginations containing mitochondria, and connections to the cytoskeleton. Subsequent purification to nuclear envelopes is additionally confounded by connections of inner nuclear membrane proteins to chromatin. For these reasons, it is necessary to confirm proteomic identification of nuclear envelope proteins by testing targeting of individual proteins. The proteomic identification of nuclear envelope fractions is affected by the tendencies of transmembrane proteins to have extreme isoelectric points, strongly hydrophobic peptides, posttranslational modifications, and a propensity to aggregate, thus making proteolysis inefficient. To circumvent these problems, we have developed a MudPIT approach that uses multiple extractions and sequential proteolysis to increase identifications. Here we describe methods for isolating nuclear envelopes, determining their proteome by MudPIT, and confirming their targeting to the nuclear periphery by microscopy.

  12. Structural and functional adaptations of the mammalian nuclear envelope to meet the meiotic requirements

    PubMed Central

    Link, Jana; Jahn, Daniel; Alsheimer, Manfred

    2015-01-01

    Numerous studies in the past years provided definite evidence that the nuclear envelope is much more than just a simple barrier. It rather constitutes a multifunctional platform combining structural and dynamic features to fulfill many fundamental functions such as chromatin organization, regulation of transcription, signaling, but also structural duties like maintaining general nuclear architecture and shape. One additional and, without doubt, highly impressive aspect is the recently identified key function of selected nuclear envelope components in driving meiotic chromosome dynamics, which in turn is essential for accurate recombination and segregation of the homologous chromosomes. Here, we summarize the recent work identifying new key players in meiotic telomere attachment and movement and discuss the latest advances in our understanding of the actual function of the meiotic nuclear envelope. PMID:25674669

  13. The isolation of nuclear envelopes. Effects of thiol-group oxidation and of calcium ions.

    PubMed

    Comerford, S A; McLuckie, I F; Gorman, M; Scott, K A; Agutter, P S

    1985-02-15

    The effects of (a) oxidative cross-linking of protein thiol groups and (b) the presence or absence of Ca2+ ions on rat liver nuclear-envelope isolation were studied. Two envelope-isolation procedures were compared: a well characterized low-ionic-strength method and a recently developed high-ionic-strength method. The latter method seems preferable to the former in respect of lower intranuclear contamination of the envelopes, suppression of endogenous serine proteinase, and maintenance of high specific activities of envelope-associated enzymes. In both procedures, however, the presence of Ca2+ gave rise to a rapid, apparently irreversible, contamination of the envelopes by intranuclear material. This effect was half-maximal at 20 microM-Ca2+. In addition, the envelopes became contaminated with intranuclear material by a Ca2+-independent mechanism, apparently resulting from N-ethylmaleimide-sensitive intermolecular disulphide-bond formation. This oxidative process seemed to have two major kinetic components (half-life, t1/2, approx. 2 min and 10 min). In view of these findings, it is recommended that (i) for most purposes, nuclear envelopes be isolated by the newly developed high-ionic-strength procedure, (ii) irrespective of the method used, Ca2+-chelators be included in all the buffers, (iii) thiol-group oxidation be prevented or reversed during the procedure.

  14. The isolation of nuclear envelopes. Effects of thiol-group oxidation and of calcium ions.

    PubMed Central

    Comerford, S A; McLuckie, I F; Gorman, M; Scott, K A; Agutter, P S

    1985-01-01

    The effects of (a) oxidative cross-linking of protein thiol groups and (b) the presence or absence of Ca2+ ions on rat liver nuclear-envelope isolation were studied. Two envelope-isolation procedures were compared: a well characterized low-ionic-strength method and a recently developed high-ionic-strength method. The latter method seems preferable to the former in respect of lower intranuclear contamination of the envelopes, suppression of endogenous serine proteinase, and maintenance of high specific activities of envelope-associated enzymes. In both procedures, however, the presence of Ca2+ gave rise to a rapid, apparently irreversible, contamination of the envelopes by intranuclear material. This effect was half-maximal at 20 microM-Ca2+. In addition, the envelopes became contaminated with intranuclear material by a Ca2+-independent mechanism, apparently resulting from N-ethylmaleimide-sensitive intermolecular disulphide-bond formation. This oxidative process seemed to have two major kinetic components (half-life, t1/2, approx. 2 min and 10 min). In view of these findings, it is recommended that (i) for most purposes, nuclear envelopes be isolated by the newly developed high-ionic-strength procedure, (ii) irrespective of the method used, Ca2+-chelators be included in all the buffers, (iii) thiol-group oxidation be prevented or reversed during the procedure. PMID:2983687

  15. Differential localization of 5- and 15-lipoxygenases to the nuclear envelope in RAW macrophages.

    PubMed

    Christmas, P; Fox, J W; Ursino, S R; Soberman, R J

    1999-09-01

    Leukotriene formation is initiated in myeloid cells by an increase in intracellular calcium and translocation of 5-lipoxygenase from the cytoplasm to the nuclear envelope where it can utilize arachidonic acid. Monocyte- macrophages and eosinophils also express 15-lipoxygenase, which converts arachidonic acid to 15(S)-hydroxyeicosatetraenoic acid. Enhanced green fluorescent 5-lipoxygenase (5-LO) and 15-lipoxygenase (15-LO) fusion proteins were expressed in the cytoplasm of RAW 264.7 macrophages. Only 5-lipoxygenase translocated to the nuclear envelope after cell stimulation, suggesting that differential subcellular compartmentalization can regulate the generation of leukotrienes versus 15(S)-hydroxyeicosatetraenoic acid in cells that possess both lipoxygenases. A series of truncation mutants of 5-LO were created to identify putative targeting domains; none of these mutants localized to the nuclear envelope. The lack of targeting of 15-LO was then exploited to search for specific targeting motifs in 5-LO, by creating 5-LO/15-LO chimeric molecules. The only chimera that could sustain nuclear envelope translocation was one which involved replacement of the N-terminal 237 amino acids with the corresponding segment of 15-LO. Significantly, no discrete targeting domain could be identified in 5-LO, suggesting that sequences throughout the molecule are required for nuclear envelope localization.

  16. Nuclear pore assembly proceeds by an inside-out extrusion of the nuclear envelope

    PubMed Central

    Otsuka, Shotaro; Bui, Khanh Huy; Schorb, Martin; Hossain, M Julius; Politi, Antonio Z; Koch, Birgit; Eltsov, Mikhail; Beck, Martin; Ellenberg, Jan

    2016-01-01

    The nuclear pore complex (NPC) mediates nucleocytoplasmic transport through the nuclear envelope. How the NPC assembles into this double membrane boundary has remained enigmatic. Here, we captured temporally staged assembly intermediates by correlating live cell imaging with high-resolution electron tomography and super-resolution microscopy. Intermediates were dome-shaped evaginations of the inner nuclear membrane (INM), that grew in diameter and depth until they fused with the flat outer nuclear membrane. Live and super-resolved fluorescence microscopy revealed the molecular maturation of the intermediates, which initially contained the nuclear and cytoplasmic ring component Nup107, and only later the cytoplasmic filament component Nup358. EM particle averaging showed that the evagination base was surrounded by an 8-fold rotationally symmetric ring structure from the beginning and that a growing mushroom-shaped density was continuously associated with the deforming membrane. Quantitative structural analysis revealed that interphase NPC assembly proceeds by an asymmetric inside-out extrusion of the INM. DOI: http://dx.doi.org/10.7554/eLife.19071.001 PMID:27630123

  17. Remodeling of the Nuclear Envelope and Lamina during Bovine Preimplantation Development and Its Functional Implications.

    PubMed

    Popken, Jens; Graf, Alexander; Krebs, Stefan; Blum, Helmut; Schmid, Volker J; Strauss, Axel; Guengoer, Tuna; Zakhartchenko, Valeri; Wolf, Eckhard; Cremer, Thomas

    2015-01-01

    The present study demonstrates a major remodeling of the nuclear envelope and its underlying lamina during bovine preimplantation development. Up to the onset of major embryonic genome activation (MGA) at the 8-cell stage nuclei showed a non-uniform distribution of nuclear pore complexes (NPCs). NPCs were exclusively present at sites where DNA contacted the nuclear lamina. Extended regions of the lamina, which were not contacted by DNA, lacked NPCs. In post-MGA nuclei the whole lamina was contacted rather uniformly by DNA. Accordingly, NPCs became uniformly distributed throughout the entire nuclear envelope. These findings shed new light on the conditions which control the integration of NPCs into the nuclear envelope. The switch from maternal to embryonic production of mRNAs was accompanied by multiple invaginations covered with NPCs, which may serve the increased demands of mRNA export and protein import. Other invaginations, as well as interior nuclear segments and vesicles without contact to the nuclear envelope, were exclusively positive for lamin B. Since the abundance of these invaginations and vesicles increased in concert with a massive nuclear volume reduction, we suggest that they reflect a mechanism for fitting the nuclear envelope and its lamina to a shrinking nuclear size during bovine preimplantation development. In addition, a deposit of extranuclear clusters of NUP153 (a marker for NPCs) without associated lamin B was frequently observed from the zygote stage up to MGA. Corresponding RNA-Seq data revealed deposits of spliced, maternally provided NUP153 mRNA and little unspliced, newly synthesized RNA prior to MGA, which increased strongly at the initiation of embryonic expression of NUP153 at MGA.

  18. Mechanical and molecular basis for the symmetrical division of the fission yeast nuclear envelope.

    PubMed

    Castagnetti, Stefania; Božič, Bojan; Svetina, Saša

    2015-06-28

    In fission yeast Schizosaccharomyces pombe, the nuclear envelope remains intact throughout mitosis and undergoes a series of symmetrical morphological changes when the spindle pole bodies (SPBs), embedded in the nuclear envelope, are pushed apart by elongating spindle microtubules. These symmetrical membrane shape transformations do not correspond to the shape behavior of an analogous system based on lipid vesicles. Here we report that the symmetry of the dividing fission yeast nucleus is ensured by SPB-chromosome attachments, as loss of kinetochore clustering in the vicinity of SPBs results in the formation of abnormal asymmetric shapes with long membrane tethers. We integrated these findings in a biophysical model, which explains the symmetry of the nuclear shapes on the basis of forces exerted by chromosomes clustered at SPBs on the extending nuclear envelope. Based on this analysis we conclude that the fission yeast nuclear envelope exhibits the same mechanical properties as simple lipid vesicles, but interactions with other cellular components, such as chromosomes, influence the nuclear shape during mitosis, allowing the formation of otherwise energetically unfavorable symmetrical dumbbell structures upon spindle elongation. The model allows us to explain the appearance of abnormal asymmetric shapes in fission yeast mutants with mis-segregated chromosomes as well as with altered nuclear membrane composition.

  19. Nuclear envelope-associated endosomes deliver surface proteins to the nucleus.

    PubMed

    Chaumet, Alexandre; Wright, Graham D; Seet, Sze Hwee; Tham, Keit Min; Gounko, Natalia V; Bard, Frederic

    2015-01-01

    Endocytosis directs molecular cargo along three main routes: recycling to the cell surface, transport to the Golgi apparatus or degradation in endolysosomes. Pseudomonas exotoxin A (PE) is a bacterial protein that typically traffics to the Golgi and then the endoplasmic reticulum before translocating to the cytosol. Here we show that a substantial fraction of internalized PE is also located in nuclear envelope-associated endosomes (NAE), which display limited mobility, exhibit a propensity to undergo fusion and readily discharge their contents into the nuclear envelope. Electron microscopy and protein trapping in the nucleus indicate that NAE mediate PE transfer into the nucleoplasm. RNAi screening further revealed that NAE-mediated transfer depends on the nuclear envelope proteins SUN1 and SUN2, as well as the Sec61 translocon complex. These data reveal a novel endosomal route from the cell surface to the nucleoplasm that facilitates the accumulation of extracellular and cell surface proteins in the nucleus. PMID:26356418

  20. Poly(A) binding proteins located at the inner surface of resealed nuclear envelopes.

    PubMed

    Prochnow, D; Riedel, N; Agutter, P S; Fasold, H

    1990-04-25

    We have used a photoreactive cross-linking reagent, poly(A/8-N3-A) (a poly(A) of average molecular mass of 100 kDa in which 5-10% of the A residues are replaced by 8-N3-A), to label poly(A) binding proteins of rat liver nuclear envelopes. This reagent was prepared by polymerizing a mixture of ADP and 8-N3-ADP with polynucleotide phosphorylase. The purified poly(A) was labeled in the 5'-position with a 32P group. In nuclear envelopes prepared by a low salt DNase I procedure, the poly(A/8-N3-A) labeled a protein-nucleic acid complex of approximately 270 kDa, which on degradation with RNase U2 or NaOH at pH 10 yielded two polypeptides of approximately 50 and 30 kDa. These photoreaction products were markedly decreased when resealed nuclear envelopes or non-nuclear envelope proteins were irradiated in the presence of poly(A/8-N3-A). The affinity labeling was intensified when resealed vesicles were made leaky by freezing or ultrasonication, suggesting that the poly(A) binding proteins are accessible from the nucleoplasmic but not the cytoplasmic face of the envelope. Moreover binding was specific for poly(A). Alternative reagents, random poly(A/8-N3-A,C,G,U) of about 100 kDa and poly(dA) (molecular mass between 350 and 515 kDa), showed a very low affinity for poly(A) recognition proteins in the low salt DNase I-treated nuclear envelopes; the 270-kDa band was labeled only weakly. The binding site was not protected by poly(A,C,G,U), weakly by poly(dA), and distinctly by poly(A).

  1. Prm3p is a pheromone-induced peripheral nuclear envelope protein required for yeast nuclear fusion.

    PubMed

    Shen, Shu; Tobery, Cynthia E; Rose, Mark D

    2009-05-01

    Nuclear membrane fusion is the last step in the mating pathway of the yeast Saccharomyces cerevisiae. We adapted a bioinformatics approach to identify putative pheromone-induced membrane proteins potentially required for nuclear membrane fusion. One protein, Prm3p, was found to be required for nuclear membrane fusion; disruption of PRM3 caused a strong bilateral defect, in which nuclear congression was completed but fusion did not occur. Prm3p was localized to the nuclear envelope in pheromone-responding cells, with significant colocalization with the spindle pole body in zygotes. A previous report, using a truncated protein, claimed that Prm3p is localized to the inner nuclear envelope. Based on biochemistry, immunoelectron microscopy and live cell microscopy, we find that functional Prm3p is a peripheral membrane protein exposed on the cytoplasmic face of the outer nuclear envelope. In support of this, mutations in a putative nuclear localization sequence had no effect on full-length protein function or localization. In contrast, point mutations and deletions in the highly conserved hydrophobic carboxy-terminal domain disrupted both protein function and localization. Genetic analysis, colocalization, and biochemical experiments indicate that Prm3p interacts directly with Kar5p, suggesting that nuclear membrane fusion is mediated by a protein complex.

  2. TMEM120A and B: Nuclear Envelope Transmembrane Proteins Important for Adipocyte Differentiation

    PubMed Central

    Batrakou, Dzmitry G.; de las Heras, Jose I.; Czapiewski, Rafal; Mouras, Rabah; Schirmer, Eric C.

    2015-01-01

    Recent work indicates that the nuclear envelope is a major signaling node for the cell that can influence tissue differentiation processes. Here we present two nuclear envelope trans-membrane proteins TMEM120A and TMEM120B that are paralogs encoded by the Tmem120A and Tmem120B genes. The TMEM120 proteins are expressed preferentially in fat and both are induced during 3T3-L1 adipocyte differentiation. Knockdown of one or the other protein altered expression of several genes required for adipocyte differentiation, Gata3, Fasn, Glut4, while knockdown of both together additionally affected Pparg and Adipoq. The double knockdown also increased the strength of effects, reducing for example Glut4 levels by 95% compared to control 3T3-L1 cells upon pharmacologically induced differentiation. Accordingly, TMEM120A and B knockdown individually and together impacted on adipocyte differentiation/metabolism as measured by lipid accumulation through binding of Oil Red O and coherent anti-Stokes Raman scattering microscopy (CARS). The nuclear envelope is linked to several lipodystrophies through mutations in lamin A; however, lamin A is widely expressed. Thus it is possible that the TMEM120A and B fat-specific nuclear envelope transmembrane proteins may play a contributory role in the tissue-specific pathology of this disorder or in the wider problem of obesity. PMID:26024229

  3. Organellar proteomics: the prizes and pitfalls of opening the nuclear envelope

    PubMed Central

    Schirmer, Eric C; Gerace, Larry

    2002-01-01

    Proteomic studies have the potential to comprehensively define the composition of organelles but are limited by the organellar cross-contamination that arises during subcellular fractionation. Comparative proteomics of organellar subfractions can mitigate these problems, as demonstrated by a recent study involving the nuclear envelope. PMID:11983061

  4. Brca2-Pds5 complexes mobilize persistent meiotic recombination sites to the nuclear envelope.

    PubMed

    Kusch, Thomas

    2015-02-15

    Homologous recombination is required for reciprocal exchange between homologous chromosome arms during meiosis. Only select meiotic recombination events become chromosomal crossovers; the majority of recombination outcomes are noncrossovers. Growing evidence suggests that crossovers are repaired after noncrossovers. Here, I report that persisting recombination sites are mobilized to the nuclear envelope of Drosophila pro-oocytes during mid-pachytene. Their number correlates with the average crossover rate per meiosis. Proteomic and interaction studies reveal that the recombination mediator Brca2 associates with lamin and the cohesion factor Pds5 to secure persistent recombination sites at the nuclear envelope. In Rad51(-/-) females, all persistent DNA breaks are directed to the nuclear envelope. By contrast, a reduction of Pds5 or Brca2 levels abolishes the movement and has a negative impact on crossover rates. The data suggest that persistent meiotic DNA double-strand breaks might correspond to crossovers, which are mobilized to the nuclear envelope for their repair. The identification of Brca2-Pds5 complexes as key mediators of this process provides a first mechanistic explanation for the contribution of lamins and cohesins to meiotic recombination.

  5. Regulation of Phosphatidylinositol-specific Phospholipase C at the Nuclear Envelope in Cardiac Myocytes

    PubMed Central

    Smrcka, Alan V.

    2014-01-01

    Phosphatidylinositol 4,5 bisphosphate hydrolysis at the plasma membrane by phospholipase C is one of the major hormone regulated intracellular signaling systems. The system generates the diffusible second messenger IP3 and the membrane bound messenger diacylglycerol. Spatial regulation of this system has been thought to be through specific subcellular distributions of the IP3 receptor or PKC. As is becoming increasingly apparent, receptor-stimulated signaling systems are also found at intracellular membranes. As discussed in this issue, GPCRs have been identified at the nuclear envelope implying intracellular localization of the signaling systems that respond to GPCRs. Here we discuss the evidence for the existence of PLC signals that regulate nuclear processes, as well as the evidence for nuclear and nuclear envelope localization of PLC signaling components, and their implications for cardiac physiology and disease. PMID:25658460

  6. Localization of P-glycoprotein at the nuclear envelope of rat brain cells

    SciTech Connect

    Babakhanian, Karlo; Bendayan, Moise; Bendayan, Reina . E-mail: r.bendayan@utoronto.ca

    2007-09-21

    P-Glycoprotein is a plasma membrane drug efflux protein implicated in extrusion of cytotoxic compounds out of a cell. There is now evidence that suggests expression of this transporter at several subcellular sites, including the nucleus, mitochondria, and Golgi apparatus. This study investigated the localization and expression of P-glycoprotein at the nuclear membrane of rat brain microvessel endothelial (RBE4) and microglial (MLS-9) cell lines. Immunocytochemistry at the light and electron microscope levels using P-glycoprotein monoclonals antibodies demonstrated the localization of the protein at the nuclear envelope of RBE4 and MLS-9 cells. Western blot analysis revealed a single band of 170-kDa in purified nuclear membranes prepared from isolated nuclei of RBE4 and MLS-9 cells. These findings indicate that P-glycoprotein is expressed at the nuclear envelope of rat brain cells and suggest a role in multidrug resistance at this subcellular site.

  7. Dynamic assembly of brambleberry mediates nuclear envelope fusion during early development.

    PubMed

    Abrams, Elliott W; Zhang, Hong; Marlow, Florence L; Kapp, Lee; Lu, Sumei; Mullins, Mary C

    2012-08-01

    To accommodate the large cells following zygote formation, early blastomeres employ modified cell divisions. Karyomeres are one such modification, mitotic intermediates wherein individual chromatin masses are surrounded by nuclear envelope; the karyomeres then fuse to form a single mononucleus. We identified brambleberry, a maternal-effect zebrafish mutant that disrupts karyomere fusion, resulting in formation of multiple micronuclei. As karyomeres form, Brambleberry protein localizes to the nuclear envelope, with prominent puncta evident near karyomere-karyomere interfaces corresponding to membrane fusion sites. brambleberry corresponds to an unannotated gene with similarity to Kar5p, a protein that participates in nuclear fusion in yeast. We also demonstrate that Brambleberry is required for pronuclear fusion following fertilization in zebrafish. Our studies provide insight into the machinery required for karyomere fusion and suggest that specialized proteins are necessary for proper nuclear division in large dividing blastomeres.

  8. Specific nuclear envelope transmembrane proteins can promote the location of chromosomes to and from the nuclear periphery

    PubMed Central

    2013-01-01

    Background Different cell types have distinctive patterns of chromosome positioning in the nucleus. Although ectopic affinity-tethering of specific loci can be used to relocate chromosomes to the nuclear periphery, endogenous nuclear envelope proteins that control such a mechanism in mammalian cells have yet to be widely identified. Results To search for such proteins, 23 nuclear envelope transmembrane proteins were screened for their ability to promote peripheral localization of human chromosomes in HT1080 fibroblasts. Five of these proteins had strong effects on chromosome 5, but individual proteins affected different subsets of chromosomes. The repositioning effects were reversible and the proteins with effects all exhibited highly tissue-restricted patterns of expression. Depletion of two nuclear envelope transmembrane proteins that were preferentially expressed in liver each reduced the normal peripheral positioning of chromosome 5 in liver cells. Conclusions The discovery of nuclear envelope transmembrane proteins that can modulate chromosome position and have restricted patterns of expression may enable dissection of the functional relevance of tissue-specific patterns of radial chromosome positioning. PMID:23414781

  9. Visualizing the Spatial Relationship of the Genome with the Nuclear Envelope Using Fluorescence In Situ Hybridization.

    PubMed

    Clements, Craig S; Bikkul, Ural; Ahmed, Mai Hassan; Foster, Helen A; Godwin, Lauren S; Bridger, Joanna M

    2016-01-01

    The genome has a special relationship with the nuclear envelope in cells. Much of the genome is anchored at the nuclear periphery, tethered by chromatin binding proteins such nuclear lamins and other integral membrane proteins. Even though there are global assays such as DAM-ID or ChIP to assess what parts of the genome are associated with the nuclear envelope, it is also essential to be able to visualize regions of the genome in order to reveal their individual relationships with nuclear structures in single cells. This is executed by fluorescence in situ hybridization (FISH) in 2-dimensional flattened nuclei (2D-FISH) or 3-dimensionally preserved cells (3D-FISH) in combination with indirect immunofluorescence to reveal structural proteins. This chapter explains the protocols for 2D- and 3D-FISH in combination with indirect immunofluorescence and discusses options for image capture and analysis. Due to the nuclear envelope proteins being part of the non-extractable nucleoskeleton, we also describe how to prepare DNA halos through salt extraction and how they can be used to study genome behavior and association when combined with 2D-FISH. PMID:27147055

  10. Visualizing the Spatial Relationship of the Genome with the Nuclear Envelope Using Fluorescence In Situ Hybridization.

    PubMed

    Clements, Craig S; Bikkul, Ural; Ahmed, Mai Hassan; Foster, Helen A; Godwin, Lauren S; Bridger, Joanna M

    2016-01-01

    The genome has a special relationship with the nuclear envelope in cells. Much of the genome is anchored at the nuclear periphery, tethered by chromatin binding proteins such nuclear lamins and other integral membrane proteins. Even though there are global assays such as DAM-ID or ChIP to assess what parts of the genome are associated with the nuclear envelope, it is also essential to be able to visualize regions of the genome in order to reveal their individual relationships with nuclear structures in single cells. This is executed by fluorescence in situ hybridization (FISH) in 2-dimensional flattened nuclei (2D-FISH) or 3-dimensionally preserved cells (3D-FISH) in combination with indirect immunofluorescence to reveal structural proteins. This chapter explains the protocols for 2D- and 3D-FISH in combination with indirect immunofluorescence and discusses options for image capture and analysis. Due to the nuclear envelope proteins being part of the non-extractable nucleoskeleton, we also describe how to prepare DNA halos through salt extraction and how they can be used to study genome behavior and association when combined with 2D-FISH.

  11. Nuclear matrix, nuclear envelope and premature aging syndromes in a translational research perspective.

    PubMed

    Cau, Pierre; Navarro, Claire; Harhouri, Karim; Roll, Patrice; Sigaudy, Sabine; Kaspi, Elise; Perrin, Sophie; De Sandre-Giovannoli, Annachiara; Lévy, Nicolas

    2014-05-01

    Lamin A-related progeroid syndromes are genetically determined, extremely rare and severe. In the past ten years, our knowledge and perspectives for these diseases has widely progressed, through the progressive dissection of their pathophysiological mechanisms leading to precocious and accelerated aging, from the genes mutations discovery until therapeutic trials in affected children. A-type lamins are major actors in several structural and functional activities at the nuclear periphery, as they are major components of the nuclear lamina. However, while this is usually poorly considered, they also play a key role within the rest of the nucleoplasm, whose defects are related to cell senescence. Although nuclear shape and nuclear envelope deformities are obvious and visible events, nuclear matrix disorganization and abnormal composition certainly represent the most important causes of cell defects with dramatic pathological consequences. Therefore, lamin-associated diseases should be better referred as laminopathies instead of envelopathies, this later being too restrictive, considering neither the key structural and functional roles of soluble lamins in the entire nucleoplasm, nor the nuclear matrix contribution to the pathophysiology of lamin-associated disorders and in particular in defective lamin A processing-associated aging diseases. Based on both our understanding of pathophysiological mechanisms and the biological and clinical consequences of progeria and related diseases, therapeutic trials have been conducted in patients and were terminated less than 10 years after the gene discovery, a quite fast issue for a genetic disease. Pharmacological drugs have been repurposed and used to decrease the toxicity of the accumulated, unprocessed and truncated prelaminA in progeria. To date, none of them may be considered as a cure for progeria and these clinical strategies were essentially designed toward reducing a subset of the most dramatic and morbid features

  12. Construction of Nuclear Envelope Shape by a High-Genus Vesicle with Pore-Size Constraint

    NASA Astrophysics Data System (ADS)

    Noguchi, Hiroshi

    2016-08-01

    Nuclear pores have an approximately uniform distribution in the nuclear envelope of most living cells. Hence, the morphology of the nuclear envelope is a spherical stomatocyte with a high genus. We have investigated the morphology of high-genus vesicles under pore-size constraint using dynamically triangulated membrane simulations. Bending-energy minimization without volume or other constraints produces a circular-cage stomatocyte, where the pores are aligned in a circular line on an oblate bud. As the pore radius is reduced, the circular-pore alignment is more stabilized than a random pore distribution on a spherical bud. However, we have clarified the conditions for the formation of a spherical stomatocyte: a small perinuclear volume, osmotic pressure within nucleoplasm, and repulsion between the pores. When area-difference elasticity is taken into account, the formation of cylindrical or budded tubules from the stomatocyte and discoidal stomatocyte is found.

  13. Construction of Nuclear Envelope Shape by a High-Genus Vesicle with Pore-Size Constraint.

    PubMed

    Noguchi, Hiroshi

    2016-08-23

    Nuclear pores have an approximately uniform distribution in the nuclear envelope of most living cells. Hence, the morphology of the nuclear envelope is a spherical stomatocyte with a high genus. We have investigated the morphology of high-genus vesicles under pore-size constraint using dynamically triangulated membrane simulations. Bending-energy minimization without volume or other constraints produces a circular-cage stomatocyte, where the pores are aligned in a circular line on an oblate bud. As the pore radius is reduced, the circular-pore alignment is more stabilized than a random pore distribution on a spherical bud. However, we have clarified the conditions for the formation of a spherical stomatocyte: a small perinuclear volume, osmotic pressure within nucleoplasm, and repulsion between the pores. When area-difference elasticity is taken into account, the formation of cylindrical or budded tubules from the stomatocyte and discoidal stomatocyte is found. PMID:27558725

  14. Inner nuclear envelope protein SUN1 plays a prominent role in mammalian mRNA export

    PubMed Central

    Li, Ping; Noegel, Angelika A.

    2015-01-01

    Nuclear export of messenger ribonucleoproteins (mRNPs) through the nuclear pore complex (NPC) can be roughly classified into two forms: bulk and specific export, involving an nuclear RNA export factor 1 (NXF1)-dependent pathway and chromosome region maintenance 1 (CRM1)-dependent pathway, respectively. SUN proteins constitute the inner nuclear envelope component of the linker of nucleoskeleton and cytoskeleton (LINC) complex. Here, we show that mammalian cells require SUN1 for efficient nuclear mRNP export. The results indicate that both SUN1 and SUN2 interact with heterogeneous nuclear ribonucleoprotein (hnRNP) F/H and hnRNP K/J. SUN1 depletion inhibits the mRNP export, with accumulations of both hnRNPs and poly(A)+RNA in the nucleus. Leptomycin B treatment indicates that SUN1 functions in mammalian mRNA export involving the NXF1-dependent pathway. SUN1 mediates mRNA export through its association with mRNP complexes via a direct interaction with NXF1. Additionally, SUN1 associates with the NPC through a direct interaction with Nup153, a nuclear pore component involved in mRNA export. Taken together, our results reveal that the inner nuclear envelope protein SUN1 has additional functions aside from being a central component of the LINC complex and that it is an integral component of the mammalian mRNA export pathway suggesting a model whereby SUN1 recruits NXF1-containing mRNP onto the nuclear envelope and hands it over to Nup153. PMID:26476453

  15. Effect of colchicine on mammalian liver nuclear envelope and on nucleo-cytoplasmic RNA transport.

    PubMed

    Agutter, P S; Suckling, K E

    1982-09-27

    The binding of colchicine to nuclear envelopes was studied in order to elucidate the mechanism whereby this compound inhibits nucleocytoplasmic RNA transport. The results suggest that a single class of colchicine-binding site (dissociation constant=approx. 0.7 mM, concentration=approx. 330 nmol colchicine/mg protein) is localised in the nuclear periphery (pore-lamina) and that binding to these sites effects a constriction of the pore-complexes with concomitant inhibition of RNA egress and disordering of the nuclear membrane phospholipid bilayers.

  16. Till disassembly do us part: a happy marriage of nuclear envelope and chromatin.

    PubMed

    Tsuchiya, Yuichi

    2008-02-01

    A characteristic feature of eukaryotic cells is the presence of nuclear envelope (NE) which separates genomic DNA from cytoplasm. NE is composed of inner nuclear membrane (INM), which interacts with chromatin, and outer nuclear membrane, which is connected to endoplasmic reticulum. Nuclear pore complexes are inserted into NE to form transport channels between nucleus and cytoplasm. In metazoan cells, an intermediate filament-based meshwork called as nuclear lamina exists between INM and chromatin. Sophisticated collaboration of these molecular machineries is necessary for the structure and functions of NE. Recent research advances have revealed that NE dynamically communicates with chromatin and cytoskeleton to control multiple nuclear functions. In this mini review, I briefly summarize the basic concepts and current topics of functional relationships between NE and chromatin.

  17. Nuclear Envelope Protein SUN2 Promotes Cyclophilin-A-Dependent Steps of HIV Replication

    PubMed Central

    Lahaye, Xavier; Satoh, Takeshi; Gentili, Matteo; Cerboni, Silvia; Silvin, Aymeric; Conrad, Cécile; Ahmed-Belkacem, Abdelhakim; Rodriguez, Elisa C.; Guichou, Jean-François; Bosquet, Nathalie; Piel, Matthieu; Le Grand, Roger; King, Megan C.; Pawlotsky, Jean-Michel; Manel, Nicolas

    2016-01-01

    Summary During the early phase of replication, HIV reverse transcribes its RNA and crosses the nuclear envelope while escaping host antiviral defenses. The host factor Cyclophilin A (CypA) is essential for these steps and binds the HIV capsid; however, the mechanism underlying this effect remains elusive. Here, we identify related capsid mutants in HIV-1, HIV-2, and SIVmac that are restricted by CypA. This antiviral restriction of mutated viruses is conserved across species and prevents nuclear import of the viral cDNA. Importantly, the inner nuclear envelope protein SUN2 is required for the antiviral activity of CypA. We show that wild-type HIV exploits SUN2 in primary CD4+ T cells as an essential host factor that is required for the positive effects of CypA on reverse transcription and infection. Altogether, these results establish essential CypA-dependent functions of SUN2 in HIV infection at the nuclear envelope. PMID:27149839

  18. Mutational analyses of fs(1)Ya, an essential, developmentally regulated, nuclear envelope protein in Drosophila

    SciTech Connect

    Liu, Jun; Song, Kiwon; Wolfner, M.F.

    1995-12-01

    The fs(1)Ya protein (YA) is an essential, maternally encoded, nuclear lamina protein that is under both developmental and cell cycle control. A strong Ya mutation results in early arrest of embryos. To define the function of YA in the nuclear envelope during early embryonic development, we characterized the phenotypes of four Ya mutant alleles and determined their molecular lesions. Ya mutant embryos arrest with abnormal nuclear envelopes prior to the first mitotic division; a proportion of embryos from two leaky Ya mutants proceed beyond this but arrest after several abnormal divisions. Ya unfertilized eggs contain nuclei of different sizes and condensation states, apparently due to abnormal fusion of the meiotic products immediately after meiosis. Lamin is localized at the periphery of the uncondensed nuclei in these eggs. These results suggest that Ya function is required during and after egg maturation to facilitate proper chromatin condensation, rather than to allow a lamin-containing nuclear envelope to form. Two leaky Ya alleles that partially complement have lesions at opposite ends of the YA protein, suggesting that the N- and C-termini are important for YA function might interact with itself either directly or indirectly. 27 refs., 6 figs.

  19. Reduction of a 4q35-encoded nuclear envelope protein in muscle differentiation

    SciTech Connect

    Ostlund, Cecilia; Guan, Tinglu; Figlewicz, Denise A.; Hays, Arthur P.; Worman, Howard J.; Gerace, Larry; Schirmer, Eric C.

    2009-11-13

    Muscular dystrophy and peripheral neuropathy have been linked to mutations in genes encoding nuclear envelope proteins; however, the molecular mechanisms underlying these disorders remain unresolved. Nuclear envelope protein p19A is a protein of unknown function encoded by a gene at chromosome 4q35. p19A levels are significantly reduced in human muscle as cells differentiate from myoblasts to myotubes; however, its levels are not similarly reduced in all differentiation systems tested. Because 4q35 has been linked to facioscapulohumeral muscular dystrophy (FSHD) and some adjacent genes are reportedly misregulated in the disorder, levels of p19A were analyzed in muscle samples from patients with FSHD. Although p19A was increased in most cases, an absolute correlation was not observed. Nonetheless, p19A downregulation in normal muscle differentiation suggests that in the cases where its gene is inappropriately re-activated it could affect muscle differentiation and contribute to disease pathology.

  20. AIM2 inflammasome is activated by pharmacological disruption of nuclear envelope integrity

    PubMed Central

    Di Micco, Antonia; Frera, Gianluca; Lugrin, Jérôme; Jamilloux, Yvan; Hsu, Erh-Ting; Tardivel, Aubry; De Gassart, Aude; Zaffalon, Léa; Bujisic, Bojan; Siegert, Stefanie; Quadroni, Manfredo; Broz, Petr; Henry, Thomas; Hrycyna, Christine A.

    2016-01-01

    Inflammasomes are critical sensors that convey cellular stress and pathogen presence to the immune system by activating inflammatory caspases and cytokines such as IL-1β. The nature of endogenous stress signals that activate inflammasomes remains unclear. Here we show that an inhibitor of the HIV aspartyl protease, Nelfinavir, triggers inflammasome formation and elicits an IL-1R–dependent inflammation in mice. We found that Nelfinavir impaired the maturation of lamin A, a structural component of the nuclear envelope, thereby promoting the release of DNA in the cytosol. Moreover, deficiency of the cytosolic DNA-sensor AIM2 impaired Nelfinavir-mediated inflammasome activation. These findings identify a pharmacologic activator of inflammasome and demonstrate the role of AIM2 in detecting endogenous DNA release upon perturbation of nuclear envelope integrity. PMID:27462105

  1. [The role of chromosomal regions anchored to the nuclear envelope in the functional organization of chromosomes].

    PubMed

    Shabarina, A N; Shostak, N G; Glazkov, M V

    2010-09-01

    The functional characteristics of the DNA fragments responsible for chromosome attachment to the nuclear envelope during the interphase (neDNAs) have been studied. The neDNAs flanking the transgene have been found to promote a steadily high rate of its expression, irrespective of the site of its insertion into the host chromosomes. At the same time, neDNAs themselves have no transcription regulatory functions. PMID:21061611

  2. A Network of Nuclear Envelope Membrane Proteins Linking Centromeres to Microtubules

    PubMed Central

    King, Megan C.; Drivas, Theodore G.; Blobel, Günter

    2008-01-01

    Summary In the fission yeast S. pombe, nuclei are actively positioned at the cell center by microtubules. Here we show that cytoplasmic microtubules are mechanically coupled to the nuclear heterochromatin through proteins embedded in the nuclear envelope. This includes an integral outer nuclear membrane protein of the KASH family (Kms2) and two integral inner nuclear membrane proteins: the SUN-domain protein Sad1 and the novel, conserved protein Ima1. Ima1 specifically binds to heterochromatic regions and promotes the tethering of centromeric DNA to the SUN-KASH complex. In the absence of Ima1, or in cells harboring mutations in the centromeric Ndc80 complex, inefficient coupling of centromeric heterochromatin to Sad1 leads to striking defects in the ability of the nucleus to tolerate microtubule-dependent forces, leading to changes in nuclear shape, loss of spindle pole body components from the nuclear envelope, and partial dissociation of SUN-KASH complexes. This work highlights a novel means of communication between cytoplasmic microtubules and chromatin. PMID:18692466

  3. This bud's for you: mechanisms of cellular nucleocytoplasmic trafficking via nuclear envelope budding.

    PubMed

    Fradkin, Lee G; Budnik, Vivian

    2016-08-01

    The nuclear envelope (NE) physically separates the cytoplasmic and nuclear compartments. While this barrier provides advantages, it also presents a challenge for the nuclear export of large ribonucleoprotein (RNP) complexes. Decades-old dogma holds that all such border-crossing is via the nuclear pore complex (NPC). However, the diameter of the NPC central channel limits the passage of large cargos. Here, we review evidence that such large RNPs employ an endogenous NE-budding pathway, previously thought to be exclusive to the nuclear egress of Herpes viruses. We discuss this and other models proposed, the likelihood that this pathway is conserved, and the consequences of disrupting NE-budding for synapse development, localized translation of synaptic mRNAs, and laminopathies inducing accelerated aging. PMID:27236823

  4. Modeling meiotic chromosome pairing: nuclear envelope attachment, telomere-led active random motion, and anomalous diffusion

    NASA Astrophysics Data System (ADS)

    Marshall, Wallace F.; Fung, Jennifer C.

    2016-04-01

    The recognition and pairing of homologous chromosomes during meiosis is a complex physical and molecular process involving a combination of polymer dynamics and molecular recognition events. Two highly conserved features of meiotic chromosome behavior are the attachment of telomeres to the nuclear envelope and the active random motion of telomeres driven by their interaction with cytoskeletal motor proteins. Both of these features have been proposed to facilitate the process of homolog pairing, but exactly what role these features play in meiosis remains poorly understood. Here we investigate the roles of active motion and nuclear envelope tethering using a Brownian dynamics simulation in which meiotic chromosomes are represented by a Rouse polymer model subjected to tethering and active forces at the telomeres. We find that tethering telomeres to the nuclear envelope slows down pairing relative to the rates achieved by unattached chromosomes, but that randomly directed active forces applied to the telomeres speed up pairing dramatically in a manner that depends on the statistical properties of the telomere force fluctuations. The increased rate of initial pairing cannot be explained by stretching out of the chromosome conformation but instead seems to correlate with anomalous diffusion of sub-telomeric regions.

  5. Stage-dependent remodeling of the nuclear envelope and lamina during rabbit early embryonic development

    PubMed Central

    POPKEN, Jens; SCHMID, Volker J.; STRAUSS, Axel; GUENGOER, Tuna; WOLF, Eckhard; ZAKHARTCHENKO, Valeri

    2015-01-01

    Utilizing 3D structured illumination microscopy, we investigated the quality and quantity of nuclear invaginations and the distribution of nuclear pores during rabbit early embryonic development and identified the exact time point of nucleoporin 153 (NUP153) association with chromatin during mitosis. Contrary to bovine early embryonic nuclei, featuring almost exclusively nuclear invaginations containing a small volume of cytoplasm, nuclei in rabbit early embryonic stages show additionally numerous invaginations containing a large volume of cytoplasm. Small-volume invaginations frequently emanated from large-volume nuclear invaginations but not vice versa, indicating a different underlying mechanism. Large- and small-volume nuclear envelope invaginations required the presence of chromatin, as they were restricted to chromatin-positive areas. The chromatin-free contact areas between nucleolar precursor bodies (NPBs) and large-volume invaginations were free of nuclear pores. Small-volume invaginations were not in contact with NPBs. The number of invaginations and isolated intranuclear vesicles per nucleus peaked at the 4-cell stage. At this stage, the nuclear surface showed highly concentrated clusters of nuclear pores surrounded by areas free of nuclear pores. Isolated intranuclear lamina vesicles were usually NUP153 negative. Cytoplasmic, randomly distributed NUP153-positive clusters were highly abundant at the zygote stage and decreased in number until they were almost absent at the 8-cell stage and later. These large NUP153 clusters may represent a maternally provided NUP153 deposit, but they were not visible as clusters during mitosis. Major genome activation at the 8- to 16-cell stage may mark the switch from a necessity for a deposit to on-demand production. NUP153 association with chromatin is initiated during metaphase before the initiation of the regeneration of the lamina. To our knowledge, the present study demonstrates for the first time major remodeling

  6. Relationship between RNA synthesis and the Ca2+-filled state of the nuclear envelope store.

    PubMed

    Benech, Juan Claudio; Escande, Carlos; Sotelo, José Roberto

    2005-08-01

    RNA synthesis and ATP-dependent (45)Ca(2+) uptake were measured simultaneously in isolated nuclear fraction of rat liver nuclei. Maximal level of RNA synthesis was obtained under ATP-dependent (45)Ca(2+)-uptake conditions (1 microM free [Ca(2+)] and 1 mM ATP in the bathing solution). This experimental condition was defined as "stimulated nuclei" condition. ATP-dependent (45)Ca(2+) uptake was inhibited using different strategies including: (a) eliminating Ca(2+) (1 mM EGTA); (b) lowering the ATP concentration; (c) modifying nuclear envelope membranes Ca(2+) permeability (Ca(2+) ionophores); or (d) inhibiting the nuclear Ca(2+) pump (thapsigargin and 3',3'',5',5''-tetraiodophenolsulfonephthalein). Under all the above conditions, RNA synthesis was lower than in "stimulated nuclei" condition. In the presence of ionomycin, RNA synthesis was significantly higher at 500 nM free [Ca(2+)], as compared with RNA synthesis in a Ca(2+)-free medium or at 1muM free [Ca(2+)]. However, even in such condition (500 nM free [Ca(2+)]), RNA synthesis was lower than RNA synthesis obtained in "stimulated nuclei" condition. We suggest two components for the effect of Ca(2+) on RNA synthesis: (A) a direct effect of nucleoplasmic [Ca(2+)]; and (B) an effect dependent on the accumulation of Ca(2+) in the nuclear envelope store mediated by the SERCA nuclear Ca(2+) pump.

  7. Cdk1 Activates Pre-mitotic Nuclear Envelope Dynein Recruitment and Apical Nuclear Migration in Neural Stem Cells.

    PubMed

    Baffet, Alexandre D; Hu, Daniel J; Vallee, Richard B

    2015-06-22

    Dynein recruitment to the nuclear envelope is required for pre-mitotic nucleus-centrosome interactions in nonneuronal cells and for apical nuclear migration in neural stem cells. In each case, dynein is recruited to the nuclear envelope (NE) specifically during G2 via two nuclear pore-mediated mechanisms involving RanBP2-BicD2 and Nup133-CENP-F. The mechanisms responsible for cell-cycle control of this behavior are unknown. We now find that Cdk1 serves as a direct master controller for NE dynein recruitment in neural stem cells and HeLa cells. Cdk1 phosphorylates conserved sites within RanBP2 and activates BicD2 binding and early dynein recruitment. Late recruitment is triggered by a Cdk1-induced export of CENP-F from the nucleus. Forced NE targeting of BicD2 overrides Cdk1 inhibition, fully rescuing dynein recruitment and nuclear migration in neural stem cells. These results reveal how NE dynein recruitment is cell-cycle regulated and identify the trigger mechanism for apical nuclear migration in the brain.

  8. Isolation of chromatin DNA tightly bound to the nuclear envelope of HeLa cells.

    PubMed

    Kuvichkin, Vasily Vladimirovich

    2012-11-01

    Recent discovery of the role of nuclear pores in transcription, predicted by our early DNA-membrane complex (DMC) model, makes membrane-bound DNA (MBD) isolation from the cell nucleus and analysis of the MBD actual. The method of MBD isolation proposed by us retains DMC integrity during isolation. We used HeLa cells for DMC extraction. Changing the ionic composition of the isolation medium and replacing DNase I, used commonly for chromatin destruction, with a set of restriction enzymes allowed us to isolate the MBD. Treatment of a nuclear membrane with proteinase K and ultrasound has been used to increase the yield of MBD. Electron microscopic analysis of the purified fraction of isolated DMC supports our previous model of nuclear envelope lipid-chromatin interaction in the nuclear pore assembly.

  9. In Situ Detection of Interactions Between Nuclear Envelope Proteins and Partners.

    PubMed

    Barateau, Alice; Buendia, Brigitte

    2016-01-01

    Proximity ligation assay (PLA) appears as a quick and easy technique to visualize within fixed cells the occurrence and in situ distribution of protein complexes. PLA has been validated to detect protein-protein interactions within the nuclear compartment. Here, we describe a protocol which allows the detection of interactions between A-type nuclear lamins and either LEM-domain proteins (such as emerin, integrated within the inner nuclear membrane, and LAP2α which accumulates within the nucleoplasm) or gene regulatory factors (e.g., the transcription factor SREBP1). The distinct amounts and patterns of PLA signals obtained for various complexes highlight the pertinence of using PLA to reveal in situ where and to which extent nuclear envelope proteins bind specific partners. PMID:27147040

  10. Aurora B–mediated localized delays in nuclear envelope formation facilitate inclusion of late-segregating chromosome fragments

    PubMed Central

    Karg, Travis; Warecki, Brandt; Sullivan, William

    2015-01-01

    To determine how chromosome segregation is coordinated with nuclear envelope formation (NEF), we examined the dynamics of NEF in the presence of lagging acentric chromosomes in Drosophila neuroblasts. Acentric chromosomes often exhibit delayed but ultimately successful segregation and incorporation into daughter nuclei. However, it is unknown whether these late-segregating acentric fragments influence NEF to ensure their inclusion in daughter nuclei. Through live analysis, we show that acentric chromosomes induce highly localized delays in the reassembly of the nuclear envelope. These delays result in a gap in the nuclear envelope that facilitates the inclusion of lagging acentrics into telophase daughter nuclei. Localized delays of nuclear envelope reassembly require Aurora B kinase activity. In cells with reduced Aurora B activity, there is a decrease in the frequency of local nuclear envelope reassembly delays, resulting in an increase in the frequency of acentric-bearing, lamin-coated micronuclei. These studies reveal a novel role of Aurora B in maintaining genomic integrity by promoting the formation of a passageway in the nuclear envelope through which late-segregating acentric chromosomes enter the telophase daughter nucleus. PMID:25877868

  11. The nuclear envelope and cancer: a diagnostic perspective and historical overview.

    PubMed

    de Las Heras, Jose I; Schirmer, Eric C

    2014-01-01

    Cancer has been diagnosed for millennia, but its cellular nature only began to be understood in the mid-nineteenth century when advances in microscopy allowed detailed specimen observations. It was soon noted that cancer cells often possessed nuclei that were altered in size and/or shape. This became an important criterion for cancer diagnosis that continues to be used today. The mechanisms linking nuclear abnormalities and cancer only started to be understood in the second half of the twentieth century, with the discovery of nuclear lamina composition differences in cancer cells compared to normal cells. The nuclear envelope, rather than providing a mere physical barrier between the genetic material in the nucleus and the cytoplasm, is a very important functional hub for many cellular processes. In this review we give an overview of the links between cancer biology and nuclear envelope, from the early days of microscopy until the present day's understanding of some of the molecular mechanisms behind those links. PMID:24563341

  12. Untethering the Nuclear Envelope and Cytoskeleton: Biologically Distinct Dystonias Arising from a Common Cellular Dysfunction

    PubMed Central

    Atai, Nadia A.; Ryan, Scott D.; Kothary, Rashmi; Breakefield, Xandra O.; Nery, Flávia C.

    2012-01-01

    Most cases of early onset DYT1 dystonia in humans are caused by a GAG deletion in the TOR1A gene leading to loss of a glutamic acid (ΔE) in the torsinA protein, which underlies a movement disorder associated with neuronal dysfunction without apparent neurodegeneration. Mutation/deletion of the gene (Dst) encoding dystonin in mice results in a dystonic movement disorder termed dystonia musculorum, which resembles aspects of dystonia in humans. While torsinA and dystonin proteins do not share modular domain architecture, they participate in a similar function by modulating a structural link between the nuclear envelope and the cytoskeleton in neuronal cells. We suggest that through a shared interaction with the nuclear envelope protein nesprin-3α, torsinA and the neuronal dystonin-a2 isoform comprise a bridge complex between the outer nuclear membrane and the cytoskeleton, which is critical for some aspects of neuronal development and function. Elucidation of the overlapping roles of torsinA and dystonin-a2 in nuclear/endoplasmic reticulum dynamics should provide insights into the cellular mechanisms underlying the dystonic phenotype. PMID:22611399

  13. A unique mechanism of nuclear division in Giardia lamblia involves components of the ventral disk and the nuclear envelope.

    PubMed

    Solari, Alberto J; Rahn, Monica I; Saura, Alicia; Lujan, Hugo D

    2003-12-01

    The fine structure of the binucleate, parasitic protist Giardia lamblia during interphase and divisional stages was studied by serial thin sectioning and three-dimensional reconstructions. The earlier sign of nuclear division is the development of a few peripheral areas of densely packed chromatin directly attached to the inner nuclear envelope. An intracytoplasmic sheet of ventral disk components grows from the cell periphery towards one of the nuclei, apparently constricting this nucleus, which becomes located at a ventral bulge. After the basal bodies become duplicated, a full nuclear division occurs in trophozoites, giving two pairs of parent-daughter nuclei. This full division occurs in a dorsal-ventral direction, with the resulting nuclear pairs located at the sides of the two sets of basal bodies. A new ventral disk is formed from the disk-derived sheets in the cell harboring the four nuclei. Cytokinesis is polymorphic, but at early stages is dorsal-to-dorsal. Encysting trophozoites show the development of Golgi cisternae stacks and dense, specific secretory granules. 3-D reconstructions show that cysts contain a single pair of incompletely strangled nuclei. The dividing Giardia lacks a typical, microtubular spindle either inside or outside the nuclei. The nuclear envelope seems to be the only structure involved in the final division of the parent-daughter nuclei.

  14. Nuclear envelope expansion is crucial for proper chromosomal segregation during a closed mitosis

    PubMed Central

    Takemoto, Ai; Kawashima, Shigehiro A.; Li, Juan-Juan; Jeffery, Linda; Yamatsugu, Kenzo; Elemento, Olivier; Nurse, Paul

    2016-01-01

    ABSTRACT Here, we screened a 10,371 library of diverse molecules using a drug-sensitive fission yeast strain to identify compounds which cause defects in chromosome segregation during mitosis. We identified a phosphorium-ylide-based compound Cutin-1 which inhibits nuclear envelope expansion and nuclear elongation during the closed mitosis of fission yeast, and showed that its target is the β-subunit of fatty acid synthase. A point mutation in the dehydratase domain of Fas1 conferred in vivo and in vitro resistance to Cutin-1. Time-lapse photomicrography showed that the bulk of the chromosomes were only transiently separated during mitosis, and nucleoli separation was defective. Subsequently sister chromatids re-associated leading to chromosomal mis-segregation. These segregation defects were reduced when the nuclear volume was increased and were increased when the nuclear volume was reduced. We propose that there needs to be sufficient nuclear volume to allow the nuclear elongation necessary during a closed mitosis to take place for proper chromosome segregation, and that inhibition of fatty acid synthase compromises nuclear elongation and leads to defects in chromosomal segregation. PMID:26869222

  15. Nuclear envelope expansion is crucial for proper chromosomal segregation during a closed mitosis.

    PubMed

    Takemoto, Ai; Kawashima, Shigehiro A; Li, Juan-Juan; Jeffery, Linda; Yamatsugu, Kenzo; Elemento, Olivier; Nurse, Paul

    2016-03-15

    Here, we screened a 10,371 library of diverse molecules using a drug-sensitive fission yeast strain to identify compounds which cause defects in chromosome segregation during mitosis. We identified a phosphorium-ylide-based compound Cutin-1 which inhibits nuclear envelope expansion and nuclear elongation during the closed mitosis of fission yeast, and showed that its target is the β-subunit of fatty acid synthase. A point mutation in the dehydratase domain of Fas1 conferred in vivo and in vitro resistance to Cutin-1. Time-lapse photomicrography showed that the bulk of the chromosomes were only transiently separated during mitosis, and nucleoli separation was defective. Subsequently sister chromatids re-associated leading to chromosomal mis-segregation. These segregation defects were reduced when the nuclear volume was increased and were increased when the nuclear volume was reduced. We propose that there needs to be sufficient nuclear volume to allow the nuclear elongation necessary during a closed mitosis to take place for proper chromosome segregation, and that inhibition of fatty acid synthase compromises nuclear elongation and leads to defects in chromosomal segregation.

  16. Loss of the integral nuclear envelope protein SUN1 induces alteration of nucleoli.

    PubMed

    Matsumoto, Ayaka; Sakamoto, Chiyomi; Matsumori, Haruka; Katahira, Jun; Yasuda, Yoko; Yoshidome, Katsuhide; Tsujimoto, Masahiko; Goldberg, Ilya G; Matsuura, Nariaki; Nakao, Mitsuyoshi; Saitoh, Noriko; Hieda, Miki

    2016-01-01

    A supervised machine learning algorithm, which is qualified for image classification and analyzing similarities, is based on multiple discriminative morphological features that are automatically assembled during the learning processes. The algorithm is suitable for population-based analysis of images of biological materials that are generally complex and heterogeneous. Here we used the algorithm wndchrm to quantify the effects on nucleolar morphology of the loss of the components of nuclear envelope in a human mammary epithelial cell line. The linker of nucleoskeleton and cytoskeleton (LINC) complex, an assembly of nuclear envelope proteins comprising mainly members of the SUN and nesprin families, connects the nuclear lamina and cytoskeletal filaments. The components of the LINC complex are markedly deficient in breast cancer tissues. We found that a reduction in the levels of SUN1, SUN2, and lamin A/C led to significant changes in morphologies that were computationally classified using wndchrm with approximately 100% accuracy. In particular, depletion of SUN1 caused nucleolar hypertrophy and reduced rRNA synthesis. Further, wndchrm revealed a consistent negative correlation between SUN1 expression and the size of nucleoli in human breast cancer tissues. Our unbiased morphological quantitation strategies using wndchrm revealed an unexpected link between the components of the LINC complex and the morphologies of nucleoli that serves as an indicator of the malignant phenotype of breast cancer cells. PMID:26962703

  17. Kar5p is required for multiple functions in both inner and outer nuclear envelope fusion in Saccharomyces cerevisiae.

    PubMed

    Rogers, Jason V; Rose, Mark D

    2014-12-02

    During mating in the budding yeast Saccharomyces cerevisiae, two haploid nuclei fuse via two sequential membrane fusion steps. SNAREs (i.e., soluble N-ethylmaleimide-sensitive factor attachment protein receptors) and Prm3p mediate outer nuclear membrane fusion, but the inner membrane fusogen remains unknown. Kar5p is a highly conserved transmembrane protein that localizes adjacent to the spindle pole body (SPB), mediates nuclear envelope fusion, and recruits Prm3p adjacent to the SPB. To separate Kar5p's functions, we tested localization, Prm3p recruitment, and nuclear fusion efficiency in various kar5 mutants. All domains and the conserved cysteine residues were essential for nuclear fusion. Several kar5 mutant proteins localized properly but did not mediate Prm3p recruitment; other kar5 mutant proteins localized and recruited Prm3p but were nevertheless defective for nuclear fusion, demonstrating additional functions beyond Prm3p recruitment. We identified one Kar5p domain required for SPB localization, which is dependent on the half-bridge protein Mps3p. Electron microscopy revealed a kar5 mutant that arrests with expanded nuclear envelope bridges, suggesting that Kar5p is required after outer nuclear envelope fusion. Finally, a split-GFP assay demonstrated that Kar5p localizes to both the inner and outer nuclear envelope. These insights suggest a mechanism by which Kar5p mediates inner nuclear membrane fusion.

  18. Nesprin-1{alpha} contributes to the targeting of mAKAP to the cardiac myocyte nuclear envelope

    SciTech Connect

    Pare, Genevieve C.; Easlick, Juliet L.; Mislow, John M.; McNally, Elizabeth M.; Kapiloff, Michael S. . E-mail: kapiloff@ohsu.edu

    2005-02-15

    Muscle A-kinase anchoring protein (mAKAP) is a scaffold protein found principally at the nuclear envelope of striated myocytes. mAKAP maintains a complex consisting of multiple signal transduction molecules including the cAMP-dependent protein kinase A, the ryanodine receptor calcium release channel, phosphodiesterase type 4D3, and protein phosphatase 2A. By an unknown mechanism, a domain containing spectrin repeats is responsible for targeting mAKAP to the nuclear envelope. We now demonstrate that the integral membrane protein nesprin-1{alpha} serves as a receptor for mAKAP on the nuclear envelope in cardiac myocytes. Nesprin-1{alpha} is inserted into the nuclear envelope by a conserved, C-terminal, klarsicht-related transmembrane domain and forms homodimers by the binding of an amino-terminal spectrin repeat domain. Through the direct binding of the nesprin-1{alpha} amino-terminal dimerization domain to the third mAKAP spectrin repeat, nesprin-1{alpha} targets mAKAP to the nuclear envelope. In turn, overexpression of these spectrin repeat domains in myocytes can displace mAKAP from nesprin-1{alpha}.

  19. Accumulation of annexin A5 at the nuclear envelope is a biomarker of cellular aging.

    PubMed

    Klement, Karolin; Melle, Christian; Murzik, Ulrike; Diekmann, Stephan; Norgauer, Johannes; Hemmerich, Peter

    2012-07-01

    Cellular senescence is a permanent cell cycle arrest induced by short telomeres or oncogenic stress in vitro and in vivo. Because no single of the established biomarkers can reliably identify senescent cells, the application of new ones may aid the diagnosis of aged cells. Here we show that annexin A5 accumulates at the nuclear envelope during replicative and drug-induced cellular senescence in primary human fibroblasts. This new cellular aging phenotype that we have termed SA-ANX5 (senescence-associated accumulation at the nuclear envelope of annexin A5) is as efficient and quantitative as the well-established senescence-associated β-galactosidase activity assay and p21 immunoreactivity. SA-ANX5 is also observed in aged human skin where is exclusively detected in DNA damage foci-positive/Ki-67-negative cells. We also observed that depletion of annexin A5 by siRNA in human fibroblasts accelerates premature senescence through the p38MAP kinase pathway. These observations establish SA-ANX5 as a new biomarker for cellular aging and implicate a functional role for annexin A5 in cellular senescence.

  20. Transmembrane protein TMEM170A is a newly discovered regulator of ER and nuclear envelope morphogenesis in human cells

    PubMed Central

    Christodoulou, Andri; Santarella-Mellwig, Rachel; Santama, Niovi

    2016-01-01

    ABSTRACT The mechanism of endoplasmic reticulum (ER) morphogenesis is incompletely understood. ER tubules are shaped by the reticulons (RTNs) and DP1/Yop1p family members, but the mechanism of ER sheet formation is much less clear. Here, we characterize TMEM170A, a human transmembrane protein, which localizes in ER and nuclear envelope membranes. Silencing or overexpressing TMEM170A in HeLa K cells alters ER shape and morphology. Ultrastructural analysis reveals that downregulation of TMEM170A specifically induces tubular ER formation, whereas overexpression of TMEM170A induces ER sheet formation, indicating that TMEM170A is a newly discovered ER-sheet-promoting protein. Additionally, downregulation of TMEM170A alters nuclear shape and size, decreases the density of nuclear pore complexes (NPCs) in the nuclear envelope and causes either a reduction in inner nuclear membrane (INM) proteins or their relocalization to the ER. TMEM170A interacts with RTN4, a member of the reticulon family; simultaneous co-silencing of TMEM170A and RTN4 rescues ER, NPC and nuclear-envelope-related phenotypes, implying that the two proteins have antagonistic effects on ER membrane organization, and nuclear envelope and NPC formation. PMID:26906412

  1. Transmembrane protein TMEM170A is a newly discovered regulator of ER and nuclear envelope morphogenesis in human cells.

    PubMed

    Christodoulou, Andri; Santarella-Mellwig, Rachel; Santama, Niovi; Mattaj, Iain W

    2016-04-15

    The mechanism of endoplasmic reticulum (ER) morphogenesis is incompletely understood. ER tubules are shaped by the reticulons (RTNs) and DP1/Yop1p family members, but the mechanism of ER sheet formation is much less clear. Here, we characterize TMEM170A, a human transmembrane protein, which localizes in ER and nuclear envelope membranes. Silencing or overexpressing TMEM170A in HeLa K cells alters ER shape and morphology. Ultrastructural analysis reveals that downregulation of TMEM170A specifically induces tubular ER formation, whereas overexpression of TMEM170A induces ER sheet formation, indicating that TMEM170A is a newly discovered ER-sheet-promoting protein. Additionally, downregulation of TMEM170A alters nuclear shape and size, decreases the density of nuclear pore complexes (NPCs) in the nuclear envelope and causes either a reduction in inner nuclear membrane (INM) proteins or their relocalization to the ER. TMEM170A interacts with RTN4, a member of the reticulon family; simultaneous co-silencing of TMEM170A and RTN4 rescues ER, NPC and nuclear-envelope-related phenotypes, implying that the two proteins have antagonistic effects on ER membrane organization, and nuclear envelope and NPC formation.

  2. Exploring laser-induced breakdown spectroscopy for nuclear materials analysis and in-situ applications

    NASA Astrophysics Data System (ADS)

    Martin, Madhavi Z.; Allman, Steve; Brice, Deanne J.; Martin, Rodger C.; Andre, Nicolas O.

    2012-08-01

    Laser-induced breakdown spectroscopy (LIBS) has been used to determine the limits of detection of strontium (Sr) and cesium (Cs), common nuclear fission products. Additionally, detection limits were determined for cerium (Ce), often used as a surrogate for radioactive plutonium in laboratory studies. Results were obtained using a laboratory instrument with a Nd:YAG laser at fundamental wavelength of 1064 nm, frequency doubled to 532 nm with energy of 50 mJ/pulse. The data was compared for different concentrations of Sr and Ce dispersed in a CaCO3 (white) and carbon (black) matrix. We have addressed the sampling errors, limits of detection, reproducibility, and accuracy of measurements as they relate to multivariate analysis in pellets that were doped with the different elements at various concentrations. These results demonstrate that LIBS technique is inherently well suited for in situ analysis of nuclear materials in hot cells. Three key advantages are evident: (1) small samples (mg) can be evaluated; (2) nuclear materials can be analyzed with minimal sample preparation; and (3) samples can be remotely analyzed very rapidly (ms-seconds). Our studies also show that the methods can be made quantitative. Very robust multivariate models have been used to provide quantitative measurement and statistical evaluation of complex materials derived from our previous research on wood and soil samples.

  3. Exploring laser-induced breakdown spectroscopy for nuclear materials analysis and in-situ applications

    SciTech Connect

    Martin, Madhavi Z; Allman, Steve L; Brice, Deanne Jane; Martin, Rodger Carl; Andre, Nicolas O

    2012-01-01

    Laser-induced breakdown spectroscopy (LIBS) has been used to determine the limits of detection of strontium (Sr) and cesium (Cs), common nuclear fission products. Additionally, detection limits were determined for cerium (Ce), often used as a surrogate for radioactive plutonium in laboratory studies. Results were obtained using a laboratory instrument with a Nd:YAG laser at fundamental wavelength of 1064 nm, frequency doubled to 532 nm with energy of 50 mJ/pulse. The data was compared for different concentrations of Sr and Ce dispersed in a CaCO3 (white) and carbon (black) matrix. We have addressed the sampling errors, limits of detection, reproducibility, and accuracy of measurements as they relate to multivariate analysis in pellets that were doped with the different elements at various concentrations. These results demonstrate that LIBS technique is inherently well suited for in situ analysis of nuclear materials in hot cells. Three key advantages are evident: (1) small samples (mg) can be evaluated; (2) nuclear materials can be analyzed with minimal sample preparation; and (3) samples can be remotely analyzed very rapidly (ms-seconds). Our studies also show that the methods can be made quantitative. Very robust multivariate models have been used to provide quantitative measurement and statistical evaluation of complex materials derived from our previous research on wood and soil samples.

  4. Nuclear Envelope Retention of LINC Complexes Is Promoted by SUN-1 Oligomerization in the Caenorhabditis elegans Germ Line.

    PubMed

    Daryabeigi, Anahita; Woglar, Alexander; Baudrimont, Antoine; Silva, Nicola; Paouneskou, Dimitra; Vesely, Cornelia; Rauter, Manuel; Penkner, Alexandra; Jantsch, Michael; Jantsch, Verena

    2016-06-01

    SUN (Sad1 and UNC-84) and KASH (Klarsicht, ANC-1, and Syne homology) proteins are constituents of the inner and outer nuclear membranes. They interact in the perinuclear space via C-terminal SUN-KASH domains to form the linker of nucleoskeleton and cytoskeleton (LINC) complex thereby bridging the nuclear envelope. LINC complexes mediate numerous biological processes by connecting chromatin with the cytoplasmic force-generating machinery. Here we show that the coiled-coil domains of SUN-1 are required for oligomerization and retention of the protein in the nuclear envelope, especially at later stages of female gametogenesis. Consistently, deletion of the coiled-coil domain makes SUN-1 sensitive to unilateral force exposure across the nuclear membrane. Premature loss of SUN-1 from the nuclear envelope leads to embryonic death due to loss of centrosome-nuclear envelope attachment. However, in contrast to previous notions we can show that the coiled-coil domain is dispensable for functional LINC complex formation, exemplified by successful chromosome sorting and synapsis in meiotic prophase I in its absence. PMID:27098914

  5. Expression of Leukemia-Associated Nup98 Fusion Proteins Generates an Aberrant Nuclear Envelope Phenotype

    PubMed Central

    Fahrenkrog, Birthe; Martinelli, Valérie; Nilles, Nadine; Fruhmann, Gernot; Chatel, Guillaume; Juge, Sabine; Sauder, Ursula; Di Giacomo, Danika; Mecucci, Cristina; Schwaller, Jürg

    2016-01-01

    Chromosomal translocations involving the nucleoporin NUP98 have been described in several hematopoietic malignancies, in particular acute myeloid leukemia (AML). In the resulting chimeric proteins, Nup98's N-terminal region is fused to the C-terminal region of about 30 different partners, including homeodomain (HD) transcription factors. While transcriptional targets of distinct Nup98 chimeras related to immortalization are relatively well described, little is known about other potential cellular effects of these fusion proteins. By comparing the sub-nuclear localization of a large number of Nup98 fusions with HD and non-HD partners throughout the cell cycle we found that while all Nup98 chimeras were nuclear during interphase, only Nup98-HD fusion proteins exhibited a characteristic speckled appearance. During mitosis, only Nup98-HD fusions were concentrated on chromosomes. Despite the difference in localization, all tested Nup98 chimera provoked morphological alterations in the nuclear envelope (NE), in particular affecting the nuclear lamina and the lamina-associated polypeptide 2α (LAP2α). Importantly, such aberrations were not only observed in transiently transfected HeLa cells but also in mouse bone marrow cells immortalized by Nup98 fusions and in cells derived from leukemia patients harboring Nup98 fusions. Our findings unravel Nup98 fusion-associated NE alterations that may contribute to leukemogenesis. PMID:27031510

  6. Expression of Leukemia-Associated Nup98 Fusion Proteins Generates an Aberrant Nuclear Envelope Phenotype.

    PubMed

    Fahrenkrog, Birthe; Martinelli, Valérie; Nilles, Nadine; Fruhmann, Gernot; Chatel, Guillaume; Juge, Sabine; Sauder, Ursula; Di Giacomo, Danika; Mecucci, Cristina; Schwaller, Jürg

    2016-01-01

    Chromosomal translocations involving the nucleoporin NUP98 have been described in several hematopoietic malignancies, in particular acute myeloid leukemia (AML). In the resulting chimeric proteins, Nup98's N-terminal region is fused to the C-terminal region of about 30 different partners, including homeodomain (HD) transcription factors. While transcriptional targets of distinct Nup98 chimeras related to immortalization are relatively well described, little is known about other potential cellular effects of these fusion proteins. By comparing the sub-nuclear localization of a large number of Nup98 fusions with HD and non-HD partners throughout the cell cycle we found that while all Nup98 chimeras were nuclear during interphase, only Nup98-HD fusion proteins exhibited a characteristic speckled appearance. During mitosis, only Nup98-HD fusions were concentrated on chromosomes. Despite the difference in localization, all tested Nup98 chimera provoked morphological alterations in the nuclear envelope (NE), in particular affecting the nuclear lamina and the lamina-associated polypeptide 2α (LAP2α). Importantly, such aberrations were not only observed in transiently transfected HeLa cells but also in mouse bone marrow cells immortalized by Nup98 fusions and in cells derived from leukemia patients harboring Nup98 fusions. Our findings unravel Nup98 fusion-associated NE alterations that may contribute to leukemogenesis.

  7. Nuclear envelope remodeling during mouse spermiogenesis: Postmeiotic expression and redistribution of germline lamin B3

    SciTech Connect

    Schuetz, Wolfgang; Alsheimer, Manfred; Oellinger, Rupert; Benavente, Ricardo . E-mail: benavente@biozentrum.uni-wuerzburg.de

    2005-07-15

    Lamins are members of a multigene family of structural nuclear envelope (NE) proteins. Differentiated mammalian somatic cells express lamins A, C, B1, and B2. The composition and organization of the nuclear lamina of mammalian spermatogenic cells differ significantly from that of somatic cells as they express lamin B1 as well as two short germ line-specific isoforms, namely lamins B3 and C2. Here we describe in detail the expression pattern and localization of lamin B3 during mouse spermatogenesis. By combining RT-PCR, immunoblotting, and immunofluorescence microscopy, we show that lamin B3 is selectively expressed during spermiogenesis (i.e., postmeiotic stages of spermatogenesis). In round spermatids, lamin B3 is distributed in the nuclear periphery and, notably, also in the nucleoplasm. In the course of spermiogenesis, lamin B3 becomes redistributed as it concentrates progressively to the posterior pole of spermatid nuclei. Our results show that during mammalian spermiogenesis the nuclear lamina is composed of B-type isoforms only, namely the ubiquitous lamin B1 and the germline-specific lamin B3. Lamin B3 is the first example of a mammalian lamin that is selectively expressed during postmeiotic stages of spermatogenesis.

  8. The tethering of chromatin to the nuclear envelope supports nuclear mechanics

    PubMed Central

    Schreiner, Sarah M.; Koo, Peter K.; Zhao, Yao; Mochrie, Simon G. J.; King, Megan C.

    2015-01-01

    The nuclear lamina is thought to be the primary mechanical defence of the nucleus. However, the lamina is integrated within a network of lipids, proteins and chromatin; the interdependence of this network poses a challenge to defining the individual mechanical contributions of these components. Here, we isolate the role of chromatin in nuclear mechanics by using a system lacking lamins. Using novel imaging analyses, we observe that untethering chromatin from the inner nuclear membrane results in highly deformable nuclei in vivo, particularly in response to cytoskeletal forces. Using optical tweezers, we find that isolated nuclei lacking inner nuclear membrane tethers are less stiff than wild-type nuclei and exhibit increased chromatin flow, particularly in frequency ranges that recapitulate the kinetics of cytoskeletal dynamics. We suggest that modulating chromatin flow can define both transient and long-lived changes in nuclear shape that are biologically important and may be altered in disease. PMID:26074052

  9. The Use of Polyacrylamide Hydrogels to Study the Effects of Matrix Stiffness on Nuclear Envelope Properties.

    PubMed

    Minaisah, Rose-Marie; Cox, Susan; Warren, Derek T

    2016-01-01

    Matrix-derived mechanical cues influence cell proliferation, motility, and differentiation. Recent findings clearly demonstrate that the nuclear envelope (NE) adapts and remodels in response to mechanical signals, including matrix stiffness, yet a plethora of studies have been performed on tissue culture plastic or glass that have a similar stiffness to cortical bone. Using methods that allow modulation of matrix stiffness will provide further insight into the role of the NE in physiological conditions and the impact of changes in stiffness observed during ageing and disease on cellular function. In this chapter, we describe the polyacrylamide hydrogel system, which allows fabrication of hydrogels with variable stiffness to better mimic the environment experienced by cells in most tissues of the body.

  10. Importance of mammalian nuclear-envelope nucleoside triphosphatase in nucleo-cytoplasmic transport of ribonucleoproteins.

    PubMed Central

    Agutter, P S; McCaldin, B; McArdle, H J

    1979-01-01

    The nucleoside triphosphate-stimulated efflux of RNA from isolated nuclei was studied under a range of conditions, and the effects of these conditions on the process were compared with the properties of the nucleoside triphosphatase located in the pore complex. A marked similarity between the rate of efflux and the rate of nucleoside triphosphate hydrolysis was apparent, in terms of substrate specificity, sensitivity to treatment with insolubilized trypsin, kinetics and the effects of increased ionic strength and of many inhibitors. These results are taken, in view of earlier evidence, to suggest that the activity of the nucleoside triphosphatase is a prerequisite for nucleo-cytoplasmic RNA transport in vivo. There are some indications that the nuclear-envelope lipid is also involved in regulating the efflux process. PMID:229828

  11. Importance of mammalian nuclear-envelope nucleoside triphosphatase in nucleo-cytoplasmic transport of ribonucleoproteins.

    PubMed

    Agutter, P S; McCaldin, B; McArdle, H J

    1979-09-15

    The nucleoside triphosphate-stimulated efflux of RNA from isolated nuclei was studied under a range of conditions, and the effects of these conditions on the process were compared with the properties of the nucleoside triphosphatase located in the pore complex. A marked similarity between the rate of efflux and the rate of nucleoside triphosphate hydrolysis was apparent, in terms of substrate specificity, sensitivity to treatment with insolubilized trypsin, kinetics and the effects of increased ionic strength and of many inhibitors. These results are taken, in view of earlier evidence, to suggest that the activity of the nucleoside triphosphatase is a prerequisite for nucleo-cytoplasmic RNA transport in vivo. There are some indications that the nuclear-envelope lipid is also involved in regulating the efflux process.

  12. The nuclear pore complex protein Tpr is a common autoantigen in sera that demonstrate nuclear envelope staining by indirect immunofluorescence.

    PubMed

    Ou, Y; Enarson, P; Rattner, J B; Barr, S G; Fritzler, M J

    2004-05-01

    We studied the autoantigen targets of 75 human sera that had antibodies to the nuclear envelope (NE) as identified by indirect immunofluorescence (IIF) on HEp-2 cells. Several different IIF staining patterns could be identified when antibodies to different components of the nuclear membrane (NM) and nuclear pore complexes (NuPC) were identified: a smooth membrane pattern characteristic of antibodies to nuclear lamins, a punctate pattern typical of antibodies to the nuclear pore complex and more complex patterns that included antibodies to nuclear and cytoplasmic organelles. Western immunoblotting of isolated nuclear and NE proteins and immunoprecipitation of radiolabelled recombinant proteins prepared by using the full-length cDNAs of the Translocated promoter region (Tpr), gp210 and p62 were used to identify specific autoantibody targets. Fifty-two of the 75 (70%) sera bound to Tpr, 25 (33%) bound to lamins A, B or C, 15 (20%) reacted with gp210 and none reacted with p62. Sixteen (21%) did not react with any of the NE components tested in our assays. The clinical features of 37 patients with anti-NE showed that there were 34 females and three males with an age range of 16-88 years (mean 59 years). The most frequent clinical diagnosis (9/37 = 24%) was autoimmune liver disease (ALD; two with primary biliary cirrhosis), followed by seven (19%) with systemic lupus erythematosus (SLE), four (11%) with a motor and/or sensory neuropathy, three (8%) with anti-phospholipid syndrome (APS), two with systemic sclerosis (SSc), two with Sjögren's syndrome (SjS), and others with a variety of diagnoses. This report indicates that Tpr, a component of the NuPC, is a common target of human autoantibodies that react with the NE.

  13. PLCgamma is enriched on poly-phosphoinositide-rich vesicles to control nuclear envelope assembly.

    PubMed

    Byrne, Richard D; Garnier-Lhomme, Marie; Han, Kevin; Dowicki, Michael; Michael, Nick; Totty, Nick; Zhendre, Vanessa; Cho, Aeri; Pettitt, Trevor R; Wakelam, Michael J; Poccia, Dominic L; Larijani, Banafshé

    2007-05-01

    Nuclear envelope assembly is an essential event in each cell cycle but the proteins and lipids involved in its regulation remain mostly unknown. Assembly involves membrane fusions but neither specific SNAREs nor Rab GTPases have been identified in its control. We report that a precursor membrane population (MV1) required for NE assembly has a unique lipid composition consisting prominently of poly-phosphatidylinositides. The lipid composition was determined by adapting HPLC electrospray ionisation tandem mass spectrometry to phosphoinositide analysis, revealing the capacity of this technique to document dynamic lipid transitions of functional importance in natural membrane populations. MV1 is >100-fold enriched in endogenous PLCgamma and >25-fold enriched in the PLC substrate phosphatidylinositol bisphosphate (PtdInsP2) compared to the second membrane population, derived largely from endoplasmic reticulum (ER), that contributes most of the NE. During NE formation PLCgamma becomes transiently phosphorylated at the tyrosine 783 site indicative of its activation. In addition specific inhibition of PLCgamma blocks nuclear envelope formation. In vivo, PLCgamma is concentrated on vesicles of similar size to purified MV1. These associate with nuclei during the period of NE formation and are distinct from ER membranes. The unprecedented concentration of PLCgamma and its substrate PtdInsP2 in a subset of membranes that binds to only two regions of the nucleus, and activation of PLCgamma by GTP during initial stages of NE formation provide a mechanism for temporal control of NE assembly and offer an explanation for how such a process of membrane fusion can be spatially regulated. PMID:17184973

  14. The Malleable Nature of the Budding Yeast Nuclear Envelope: Flares, Fusion, and Fenestrations.

    PubMed

    Meseroll, Rebecca A; Cohen-Fix, Orna

    2016-11-01

    In eukaryotes, the nuclear envelope (NE) physically separates nuclear components and activities from rest of the cell. The NE also provides rigidity to the nucleus and contributes to chromosome organization. At the same time, the NE is highly dynamic; it must change shape and rearrange its components during development and throughout the cell cycle, and its morphology can be altered in response to mutation and disease. Here we focus on the NE of budding yeast, Saccharomyces cerevisiae, which has several unique features: it remains intact throughout the cell cycle, expands symmetrically during interphase, elongates during mitosis and, expands asymmetrically during mitotic delay. Moreover, its NE is safely breached during mating and when large structures, such as nuclear pore complexes and the spindle pole body, are embedded into its double membrane. The budding yeast NE lacks lamins and yet the nucleus is capable of maintaining a spherical shape throughout interphase. Despite these eccentricities, studies of the budding yeast NE have uncovered interesting, and likely conserved, processes that contribute to NE dynamics. In particular, we discuss the processes that drive and enable NE expansion and the dramatic changes in the NE that lead to extensions and fenestrations. J. Cell. Physiol. 231: 2353-2360, 2016. Published 2016. This article is a U.S. Government work and is in the public domain in the USA.

  15. Intra-nuclear localization of two envelope proteins, gB and gD, of herpes simplex virus.

    PubMed

    Stannard, L M; Himmelhoch, S; Wynchank, S

    1996-01-01

    The envelopes of herpes simplex virus (HSV) particles are acquired from the inner nuclear membrane (INM) of the infected cell and virus-coded glycoproteins are present in the envelope of mature virions. Our ultrastructural study examined the process of virus envelopment and the targeting of two major viral glycoproteins, gB and gD, to the INM in HSV-infected human embryonic fibroblasts. It was shown that envelopment and transport of virus particles from the nucleus is facilitated by the formation of a dynamic tubulo-reticulum arising from the INM. Capsids were assembled in the nucleus and collected within INM tubules which protruded into the perinuclear space and thence into the cisternae of the endoplasmic reticulum (ER). Envelopment occurred by constriction and fusion of the tubular channel walls, releasing enveloped virions into the ER. Transport to the cell surface took place in membrane-bound compartments and probably followed the normal secretory pathway through the Golgi apparatus. Immunogold probes, tagged with specific monoclonal antibodies, were used to localize gB and gD during the process of virus maturation. Cytoplasmic membranes were not labelled, but probes bound inside the nucleus, mainly at sites of virus assembly. Labelling occurred on the nucleoplasmic side of the INM which surrounded capsids in the process of envelopment, but not on the outside of that membrane, although characteristic gB glycoprotein spikes were labelled on the envelopes of extracellular virus particles and on virions in trans-Golgi transport vesicles just prior to their release from the infected cell. gB was not detected on the surface of enveloped virions in the perinuclear space, or the cisternae of the ER or cis-Golgi, which suggests that the specific epitope was masked during that stage of intracellular processing. gD probes bound to virion envelopes and also to the tegument region of some particles found in both perinuclear and extracellular sites. We postulate the

  16. The budding yeast nuclear envelope adjacent to the nucleolus serves as a membrane sink during mitotic delay.

    PubMed

    Witkin, Keren L; Chong, Yolanda; Shao, Sichen; Webster, Micah T; Lahiri, Sujoy; Walters, Alison D; Lee, Brandon; Koh, Judice L Y; Prinz, William A; Andrews, Brenda J; Cohen-Fix, Orna

    2012-06-19

    The mechanisms that dictate nuclear shape are largely unknown. Here we screened the budding yeast deletion collection for mutants with abnormal nuclear shape. A common phenotype was the appearance of a nuclear extension, particularly in mutants in DNA repair and chromosome segregation genes. Our data suggest that these mutations led to the abnormal nuclear morphology indirectly, by causing a checkpoint-induced cell-cycle delay. Indeed, delaying cells in mitosis by other means also led to the appearance of nuclear extensions, whereas inactivating the DNA damage checkpoint pathway in a DNA repair mutant reduced the fraction of cells with nuclear extensions. Formation of a nuclear extension was specific to a mitotic delay, because cells arrested in S or G2 had round nuclei. Moreover, the nuclear extension always coincided with the nucleolus, while the morphology of the DNA mass remained largely unchanged. Finally, we found that phospholipid synthesis continued unperturbed when cells delayed in mitosis, and inhibiting phospholipid synthesis abolished the formation of nuclear extensions. Our data suggest a mechanism that promotes nuclear envelope expansion during mitosis. When mitotic progression is delayed, cells sequester the added membrane to the nuclear envelope associated with the nucleolus, possibly to avoid disruption of intranuclear organization.

  17. Human Cytomegalovirus Nuclear Egress Proteins Ectopically Expressed in the Heterologous Environment of Plant Cells are Strictly Targeted to the Nuclear Envelope

    PubMed Central

    Lamm, Christian E.; Link, Katrin; Wagner, Sabrina; Milbradt, Jens; Marschall, Manfred; Sonnewald, Uwe

    2016-01-01

    In all eukaryotic cells, the nucleus forms a prominent cellular compartment containing the cell’s nuclear genome. Although structurally similar, animal and plant nuclei differ substantially in details of their architecture. One example is the nuclear lamina, a layer of tightly interconnected filament proteins (lamins) underlying the nuclear envelope of metazoans. So far no orthologous lamin genes could be detected in plant genomes and putative lamin-like proteins are only poorly described in plants. To probe for potentially conserved features of metazoan and plant nuclear envelopes, we ectopically expressed the core nuclear egress proteins of human cytomegalovirus pUL50 and pUL53 in plant cells. pUL50 localizes to the inner envelope of metazoan nuclei and recruits the nuclear localized pUL53 to it, forming heterodimers. Upon expression in plant cells, a very similar localization pattern of both proteins could be determined. Notably, pUL50 is specifically targeted to the plant nuclear envelope in a rim-like fashion, a location to which coexpressed pUL53 becomes strictly corecruited from its initial nucleoplasmic distribution. Using pUL50 as bait in a yeast two-hybrid screening, the cytoplasmic re-initiation supporting protein RISP could be identified. Interaction of pUL50 and RISP could be confirmed by coexpression and coimmunoprecipitation in mammalian cells and by confocal laser scanning microscopy in plant cells, demonstrating partial pUL50-RISP colocalization in areas of the nuclear rim and other intracellular compartments. Thus, our study provides strong evidence for conserved structural features of plant and metazoan nuclear envelops and identifies RISP as a potential pUL50-interacting plant protein. PMID:26978388

  18. Human Cytomegalovirus Nuclear Egress Proteins Ectopically Expressed in the Heterologous Environment of Plant Cells are Strictly Targeted to the Nuclear Envelope.

    PubMed

    Lamm, Christian E; Link, Katrin; Wagner, Sabrina; Milbradt, Jens; Marschall, Manfred; Sonnewald, Uwe

    2016-03-10

    In all eukaryotic cells, the nucleus forms a prominent cellular compartment containing the cell's nuclear genome. Although structurally similar, animal and plant nuclei differ substantially in details of their architecture. One example is the nuclear lamina, a layer of tightly interconnected filament proteins (lamins) underlying the nuclear envelope of metazoans. So far no orthologous lamin genes could be detected in plant genomes and putative lamin-like proteins are only poorly described in plants. To probe for potentially conserved features of metazoan and plant nuclear envelopes, we ectopically expressed the core nuclear egress proteins of human cytomegalovirus pUL50 and pUL53 in plant cells. pUL50 localizes to the inner envelope of metazoan nuclei and recruits the nuclear localized pUL53 to it, forming heterodimers. Upon expression in plant cells, a very similar localization pattern of both proteins could be determined. Notably, pUL50 is specifically targeted to the plant nuclear envelope in a rim-like fashion, a location to which coexpressed pUL53 becomes strictly corecruited from its initial nucleoplasmic distribution. Using pUL50 as bait in a yeast two-hybrid screening, the cytoplasmic re-initiation supporting protein RISP could be identified. Interaction of pUL50 and RISP could be confirmed by coexpression and coimmunoprecipitation in mammalian cells and by confocal laser scanning microscopy in plant cells, demonstrating partial pUL50-RISP colocalization in areas of the nuclear rim and other intracellular compartments. Thus, our study provides strong evidence for conserved structural features of plant and metazoan nuclear envelops and identifies RISP as a potential pUL50-interacting plant protein.

  19. Structural insights into SUN-KASH complexes across the nuclear envelope

    PubMed Central

    Wang, Wenjia; Shi, Zhubing; Jiao, Shi; Chen, Cuicui; Wang, Huizhen; Liu, Guoguang; Wang, Qiang; Zhao, Yun; Greene, Mark I; Zhou, Zhaocai

    2012-01-01

    Linker of the nucleoskeleton and the cytoskeleton (LINC) complexes are composed of SUN and KASH domain-containing proteins and bridge the inner and outer membranes of the nuclear envelope. LINC complexes play critical roles in nuclear positioning, cell polarization and cellular stiffness. Previously, we reported the homotrimeric structure of human SUN2. We have now determined the crystal structure of the human SUN2-KASH complex. In the complex structure, the SUN domain homotrimer binds to three independent “hook”-like KASH peptides. The overall conformation of the SUN domain in the complex closely resembles the SUN domain in its apo state. A major conformational change involves the AA'-loop of KASH-bound SUN domain, which rearranges to form a mini β-sheet that interacts with the KASH peptide. The PPPT motif of the KASH domain fits tightly into a hydrophobic pocket on the homotrimeric interface of the SUN domain, which we termed the BI-pocket. Moreover, two adjacent protomers of the SUN domain homotrimer sandwich the KASH domain by hydrophobic interaction and hydrogen bonding. Mutations of these binding sites disrupt or reduce the association between the SUN and KASH domains in vitro. In addition, transfection of wild-type, but not mutant, SUN2 promotes cell migration in Ovcar-3 cells. These results provide a structural model of the LINC complex, which is essential for additional study of the physical and functional coupling between the cytoplasm and the nucleoplasm. PMID:22945352

  20. Vault mobility depends in part on microtubules and vaults can be recruited to the nuclear envelope

    SciTech Connect

    Zon, Arend van; Mossink, Marieke H.; Houtsmuller, Adriaan B.; Schoester, Martijn; Scheffer, George L.; Scheper, Rik J.; Sonneveld, Pieter; Wiemer, Erik A.C. . E-mail: e.wiemer@erasmusmc.nl

    2006-02-01

    Vaults are ribonucleoproteins that may function in intracellular transport processes. We investigated the intracellular distribution and dynamics of vaults in non-small cell lung cancer cells in which vaults are labeled with the green fluorescent protein. Immunofluorescence experiments showed that vaults are dispersed throughout the cytoplasm; a small fraction is found in close proximity to microtubules. Immunoprecipitation experiments corroborated these results showing co-precipitation of MVP and {beta}-tubulin. Using quantitative fluorescence-recovery after photobleaching (FRAP), we demonstrated that vault mobility over longer distances in part depends on intact microtubules; vaults moving slower when microtubules are depolymerized by nocodazole. Biochemical fractionation indicated a small fraction of MVP associated with the nucleus, however, no GFP-tagged vaults could be observed inside the nucleus. We observed an accumulation of vaults at the nuclear envelope upon treatment of cells with the protein synthesis inhibitor cycloheximide. Analysis of nucleo-cytoplasmic transport using a fluorescent substrate containing a classical NLS and NES expressed in MVP {sup +/+} and MVP {sup -/-} mouse embryonic fibroblasts indicated no differences in nuclear import/export kinetics, suggesting no role for vaults in these processes. We hypothesize that a subset of vaults moves directionally via microtubules, possibly towards the nucleus.

  1. Structural insights into SUN-KASH complexes across the nuclear envelope.

    PubMed

    Wang, Wenjia; Shi, Zhubing; Jiao, Shi; Chen, Cuicui; Wang, Huizhen; Liu, Guoguang; Wang, Qiang; Zhao, Yun; Greene, Mark I; Zhou, Zhaocai

    2012-10-01

    Linker of the nucleoskeleton and the cytoskeleton (LINC) complexes are composed of SUN and KASH domain-containing proteins and bridge the inner and outer membranes of the nuclear envelope. LINC complexes play critical roles in nuclear positioning, cell polarization and cellular stiffness. Previously, we reported the homotrimeric structure of human SUN2. We have now determined the crystal structure of the human SUN2-KASH complex. In the complex structure, the SUN domain homotrimer binds to three independent "hook"-like KASH peptides. The overall conformation of the SUN domain in the complex closely resembles the SUN domain in its apo state. A major conformational change involves the AA'-loop of KASH-bound SUN domain, which rearranges to form a mini β-sheet that interacts with the KASH peptide. The PPPT motif of the KASH domain fits tightly into a hydrophobic pocket on the homotrimeric interface of the SUN domain, which we termed the BI-pocket. Moreover, two adjacent protomers of the SUN domain homotrimer sandwich the KASH domain by hydrophobic interaction and hydrogen bonding. Mutations of these binding sites disrupt or reduce the association between the SUN and KASH domains in vitro. In addition, transfection of wild-type, but not mutant, SUN2 promotes cell migration in Ovcar-3 cells. These results provide a structural model of the LINC complex, which is essential for additional study of the physical and functional coupling between the cytoplasm and the nucleoplasm.

  2. Repo-Man Coordinates Chromosomal Reorganization with Nuclear Envelope Reassembly during Mitotic Exit

    PubMed Central

    Vagnarelli, Paola; Ribeiro, Susana; Sennels, Lau; Sanchez-Pulido, Luis; de Lima Alves, Flavia; Verheyen, Toon; Kelly, David A.; Ponting, Chris P.; Rappsilber, Juri; Earnshaw, William C.

    2011-01-01

    Summary Repo-Man targets protein phosphatase 1 γ (PP1γ) to chromatin at anaphase onset and regulates chromosome structure during mitotic exit. Here, we show that a Repo-Man:PP1 complex forms in anaphase following dephosphorylation of Repo-Man. Upon activation, the complex localizes to chromosomes and causes the dephosphorylation of histone H3 (Thr3, Ser10, and Ser28). In anaphase, Repo-Man has both catalytic and structural functions that are mediated by two separate domains. A C-terminal domain localizes Repo-Man to bulk chromatin in early anaphase. There, it targets PP1 for the dephosphorylation of histone H3 and possibly other chromosomal substrates. An N-terminal domain localizes Repo-Man to the chromosome periphery later in anaphase. There, it is responsible for the recruitment of nuclear components such as Importin β and Nup153 in a PP1-independent manner. These observations identify Repo-Man as a key factor that coordinates chromatin remodeling and early events of nuclear envelope reformation during mitotic exit. PMID:21820363

  3. Differential nuclear envelope assembly at the end of mitosis in suspension-cultured Apium graveolens cells.

    PubMed

    Kimura, Yuta; Kuroda, Chie; Masuda, Kiyoshi

    2010-04-01

    NMCP1 is a plant protein that has a long coiled-coil domain within the molecule. Newly identified NMCP2 of Daucus carota and Apium graveolens showed similar peripheral localization in the interphase nucleus, and the sequence spanning the coiled-coil domain exhibited significant similarity with the corresponding region of NMCP1. To better understand disassembly and assembly of the nuclear envelope (NE) during mitosis, subcellular distribution of NMCP1 and NMCP2 was examined using A. graveolens cells. AgNMCP1 (NMCP1 in Apium) disassembled at prometaphase, dispersed mainly within the spindle, and accumulated on segregating chromosomes, while AgNMCP2 (NMCP2 in Apium), following disassembly at prometaphase with timing similar to that of AgNMCP1, dispersed throughout the mitotic cytoplasm at metaphase and anaphase. The protein accumulated at the periphery of reforming nuclei at telophase. A probe for the endomembrane indicated that the nuclear membrane (NM) disappears at prometaphase and begins to reappear at early telophase. Growth of the NM continued after mitosis was completed. NMCP2 in the mitotic cytoplasm localized in vesicular structures that could be distinguished from the bulk endomembrane system. These results suggest that NMCP1 and NMCP2 are recruited for NE assembly in different pathways in mitosis and that NMCP2 associates with NM-derived vesicles in the mitotic cytoplasm.

  4. Muscular Dystrophy Mutations Impair the Nuclear Envelope Emerin Self-assembly Properties.

    PubMed

    Herrada, Isaline; Samson, Camille; Velours, Christophe; Renault, Louis; Östlund, Cecilia; Chervy, Pierre; Puchkov, Dmytro; Worman, Howard J; Buendia, Brigitte; Zinn-Justin, Sophie

    2015-12-18

    More than 100 genetic mutations causing X-linked Emery-Dreifuss muscular dystrophy have been identified in the gene encoding the integral inner nuclear membrane protein emerin. Most mutations are nonsense or frameshift mutations that lead to the absence of emerin in cells. Only very few cases are due to missense or short in-frame deletions. Molecular mechanisms explaining the corresponding emerin variants' loss of function are particularly difficult to identify because of the mostly intrinsically disordered state of the emerin nucleoplasmic region. We now demonstrate that this EmN region can be produced as a disordered monomer, as revealed by nuclear magnetic resonance, but rapidly self-assembles in vitro. Increases in concentration and temperature favor the formation of long curvilinear filaments with diameters of approximately 10 nm, as observed by electron microscopy. Assembly of these filaments can be followed by fluorescence through Thioflavin-T binding and by Fourier-transform Infrared spectrometry through formation of β-structures. Analysis of the assembly properties of five EmN variants reveals that del95-99 and Q133H impact filament assembly capacities. In cells, these variants are located at the nuclear envelope, but the corresponding quantities of emerin-emerin and emerin-lamin proximities are decreased compared to wild-type protein. Furthermore, variant P183H favors EmN aggregation in vitro, and variant P183T provokes emerin accumulation in cytoplasmic foci in cells. Substitution of residue Pro183 might systematically favor oligomerization, leading to emerin aggregation and mislocalization in cells. Our results suggest that emerin self-assembly is necessary for its proper function and that a loss of either the protein itself or its ability to self-assemble causes muscular dystrophy.

  5. Dystonin/Bpag1 is a necessary endoplasmic reticulum/nuclear envelope protein in sensory neurons

    SciTech Connect

    Young, Kevin G.; Kothary, Rashmi

    2008-09-10

    Dystonin/Bpag1 proteins are cytoskeletal linkers whose loss of function in mice results in a hereditary sensory neuropathy with a progressive loss of limb coordination starting in the second week of life. These mice, named dystonia musculorum (dt), succumb to the disease and die of unknown causes prior to sexual maturity. Previous evidence indicated that cytoskeletal defects in the axon are a primary cause of dt neurodegeneration. However, more recent data suggests that other factors may be equally important contributors to the disease process. In the present study, we demonstrate perikaryal defects in dorsal root ganglion (DRG) neurons at stages preceding the onset of loss of limb coordination in dt mice. Abnormalities include alterations in endoplasmic reticulum (ER) chaperone protein expression, indicative of an ER stress response. Dystonin in sensory neurons localized in association with the ER and nuclear envelope (NE). A fusion protein ofthe dystonin-a2 isoform, which harbors an N-terminal transmembrane domain, associated with and reorganized the ER in cell culture. This isoform also interacts with the NE protein nesprin-3{alpha}, but not nesprin-3{beta}. Defects in dt mice, as demonstrated here, may ultimately result in pathogenesis involving ER dysfunction and contribute significantly to the dt phenotype.

  6. Identifying Protein-Protein Associations at the Nuclear Envelope with BioID.

    PubMed

    Kim, Dae In; Jensen, Samuel C; Roux, Kyle J

    2016-01-01

    The nuclear envelope (NE) is a critical cellular structure whose constituents and roles in a myriad of cellular processes seem ever expanding. To determine the underlying mechanisms by which the NE constituents participate in various cellular events, it is necessary to understand the nature of their protein-protein associations. BioID (proximity-dependent biotin identification) is a recently established method to generate a history of protein-protein associations as they occur over time in living cells. BioID is based on fusion of a bait protein to a promiscuous biotin ligase. Expression of the BioID fusion protein in a relevant cellular environment enables biotinylation of vicinal and interacting proteins of the bait protein, permitting isolation and identification by conventional biotin-affinity capture and mass-spec analysis. In this way, BioID provides unique capabilities to identify protein-protein associations at the NE. In this chapter we provide a detailed protocol for the application of BioID to the study of NE proteins.

  7. Effects of 2-acetylaminofluorene, dietary fats and antioxidants on nuclear envelope cytochrome P-450

    SciTech Connect

    Carubelli, R.; Graham, S.A.; Griffin, M.J.; McCay, P.B.

    1986-05-01

    The authors reported a marked loss of cytochrome P-450 in hepatic nuclear envelope (NE) but not in microsomes of male Sprague-Dawley rats fed a semipurified diet containing 0.05% w/w 2-acetylaminofluorene (AAF) for 3 weeks. This may reflect loss of NE capacity to detoxify AAF metabolites generated by microsomal P-450. They are now investigating if dietary effects such as progressive decrease in the incidence of AAF-induced tumors in rats fed high polyunsaturated fat diet (HPUF) vs. high saturated fat diet (HSF) vs. low fat diet (LF), and the anticarcinogenic activity of butylated hydroxytoluene (BHT; 0.3% w/w) correlate with preservation of NE P-450. Rats fed AAF HSF (25.6% w/w corn oil) showed marked loss of NE P-450 after 3 weeks; BHT protected against this loss. Rats fed AAF in HSF (25.6% w/w; 18 parts beef tallow + 2 parts corn oil), on the other hand, experienced a marked drop in NE P-450 after 9 weeks; BHT protected against this loss. Comparison of NE P-450 levels in control rats fed HPUF or HSF for 3 weeks with those of rats fed a semipurified diet with 10% fat or Purina chow (ca. 5% fat), support the prediction of an inverse correlation between the levels of dietary fat and the NE P-450 content. Studies on AAF and BHT effects using LF (2% w/w corn oil) are in progress.

  8. Lamin Mutations Accelerate Aging via Defective Export of Mitochondrial mRNAs through Nuclear Envelope Budding.

    PubMed

    Li, Yihang; Hassinger, Linda; Thomson, Travis; Ding, Baojin; Ashley, James; Hassinger, William; Budnik, Vivian

    2016-08-01

    Defective RNA metabolism and transport are implicated in aging and degeneration [1, 2], but the underlying mechanisms remain poorly understood. A prevalent feature of aging is mitochondrial deterioration [3]. Here, we link a novel mechanism for RNA export through nuclear envelope (NE) budding [4, 5] that requires A-type lamin, an inner nuclear membrane-associated protein, to accelerated aging observed in Drosophila LaminC (LamC) mutations. These LamC mutations were modeled after A-lamin (LMNA) mutations causing progeroid syndromes (PSs) in humans. We identified mitochondrial assembly regulatory factor (Marf), a mitochondrial fusion factor (mitofusin), as well as other transcripts required for mitochondrial integrity and function, in a screen for RNAs that exit the nucleus through NE budding. PS-modeled LamC mutations induced premature aging in adult flight muscles, including decreased levels of specific mitochondrial protein transcripts (RNA) and progressive mitochondrial degradation. PS-modeled LamC mutations also induced the accelerated appearance of other phenotypes associated with aging, including a progressive accumulation of polyubiquitin aggregates [6, 7] and myofibril disorganization [8, 9]. Consistent with these observations, the mutants had progressive jumping and flight defects. Downregulating marf alone induced the above aging defects. Nevertheless, restoring marf was insufficient for rescuing the aging phenotypes in PS-modeled LamC mutations, as other mitochondrial RNAs are affected by inhibition of NE budding. Analysis of NE budding in dominant and recessive PS-modeled LamC mutations suggests a mechanism by which abnormal lamina organization prevents the egress of these RNAs via NE budding. These studies connect defects in RNA export through NE budding to progressive loss of mitochondrial integrity and premature aging. PMID:27451905

  9. Preparation and mechanism insight of nuclear envelope-like polymer vesicles for facile loading of biomacromolecules and enhanced biocatalytic activity.

    PubMed

    Zhu, Yunqing; Wang, Fangyingkai; Zhang, Cong; Du, Jianzhong

    2014-07-22

    The facile loading of sensitive and fragile biomacromolecules, such as glucose oxidase, hemoglobin, and ribonucleic acid (RNA), via synthetic vehicles directly in pure aqueous media is an important technical challenge. Inspired by the nucleus pore complex that connects the cell nucleus and the cytoplasm across the nuclear envelope, here we describe the development of a kind of polymeric nuclear envelope-like vesicle (NEV) to address this problem. The NEV is tailored to form the polymer pore complex (70 nm, similar to a nucleus pore complex) within the vesicle membrane based on nanophase segregation, which is confirmed via fluorescence spectrometry and dynamic light scattering (DLS) during self-assembly. This pH-triggered polymer pore complex can mediate the transportation of biomacromolecules across the vesicle membrane. Moreover, the NEVs facilitate the natural consecutive enzyme-catalyzed reactions via the H(+) sponge effect. This simple strategy might also be extended for mimicking other synthetic cell organelles.

  10. The fluidity of the nuclear envelope lipid does not affect the rate of nucleocytoplasmic RNA transport in mammalian liver.

    PubMed

    Agutter, P S; Suckling, K E

    1982-03-29

    The effects of in vitro and in vivo modifications of nuclear envelope lipid on DNa leakage and on ATP-stimulated RNA release from isolated rat liver nuclei were investigated. The modifications included corn-oil feeding of the animals to alter the fatty acid composition of the lipids, phospholipase treatment of the isolated nuclei, and extraction of the total lipid with Triton X-100. Significant changes in lipid composition and approximate order parameter values of the spin-label 5-doxylstearate resulted, but there was no significant effect on RNA transport rate. It was concluded that the nuclear envelope lipid does not play any important part in nucleocytoplasmic RNA transport in mammalian liver.

  11. Proteomics on the rims; insights into the biology of the nuclear envelope and flagellar pocket of trypanosomes

    PubMed Central

    Field, Mark C.; Adung’a, Vincent; Obado, Samson; Chait, Brian T.; Rout, Michael P.

    2014-01-01

    SUMMERY Trypanosomatids represent the causative agents of major diseases in humans, livestock and plants, with inevitable suffering and economic hardship as a result. They are also evolutionarily highly divergent organisms, and the many unique aspects of trypanosome biology provide opportunities in terms of identification of drug targets, the challenge of exploiting these putative targets, and at the same time significant scope for exploration of novel and divergent cell biology. We can estimate from genome sequences that the degree of divergence of trypanosomes from animals and fungi is extreme, with perhaps one third to one half of predicted trypanosome proteins having no known function based on homology or recognizable protein domains/architecture. Two highly important aspects of trypanosome biology are the flagellar pocket and the nuclear envelope, where in silico analysis clearly suggests great potential divergence in the proteome. The flagellar pocket is the sole site of endo- and exocytosis in trypanosomes and plays important roles in immune evasion via variant surface glycoprotein (VSG) trafficking and providing a location for sequestration of various invariant receptors. The trypanosome nuclear envelope has been largely unexplored, but by analogy with higher eukaryotes, roles in the regulation of chromatin and most significantly, in controlling VSG gene expression are expected. Here we discuss recent successful proteomics-based approaches towards characterization of the nuclear envelope and the endocytic apparatus, the identification of conserved and novel trypanosomatid-specific features, and the implications of these findings. PMID:22309600

  12. Effects of cholesterol on the properties of the membranes of isolated sheep liver nuclei and nuclear envelopes.

    PubMed

    Agutter, P S; Suckling, K E

    1981-04-22

    The exchangeability of cholesterol between sheep liver nuclear membranes and liposomes, and the effect of cholesterol on the fluidity of the membrane lipid were studied. In intact nuclei, the cholesterol/phospholipid ratio increased from 0.102 to 0.145 mol/mol on incubation with cholesterol-rich liposomes, with a time for half-maximal uptake of 4.2 h. In isolated envelopes under the same conditions, the ratio increased from 0.110 to 0.266 mol/mol with a time for half-maximal uptake of about 1.9 h. Moreover, the approximate order parameter of the spin label 5-(N-oxyl-4',4'-dimethyloxazolidino)-stearic acid was 0.677 in intact nuclei and 0.723 in isolated envelopes prior to exchange; after exchange, these values increased to 0.717 and 0.756, respectively. These differences between the preparations could not be attributed to differences in the capacity for cholesterol uptake between the two nuclear membranes, or to a slow rate of exchange between them; the presence of an intact nuclear matrix appeared both to disorder the lipid partially and to inhibit cholesterol uptake. The differences indicate that conclusions based on physical studies of the membrane lipid in isolated envelopes are not necessarily applicable to the intact nucleus.

  13. Functional dissection of nuclear envelope mRNA translocation system: effects of phorbol ester and a monoclonal antibody recognizing cytoskeletal structures.

    PubMed

    Schröder, H C; Diehl-Seifert, B; Rottmann, M; Messer, R; Bryson, B A; Agutter, P S; Müller, W E

    1988-03-01

    Unidirectional transport of poly(A)-containing mRNA [poly(A)+ mRNA] through the nuclear envelope pore complex is thought to be an energy (ATP or GTP)-dependent process which involves a nuclear envelope nucleoside triphosphatase (NTPase). In the intact envelope, this enzyme is regulatable by poly(A) binding and by poly(A)-dependent phosphorylation/dephosphorylation of other components of the mRNA translocation system, which are as yet unidentified. Monoclonal antibodies (mAbs) were elicited against the poly(A) binding nuclear envelope fraction isolated from rat liver. The mAbs were screened for their modulatory effects on mRNA transport in vitro. One stable clone decreased the efflux of rapidly labeled RNA and of one specific mRNA (ovalbumin) from isolated nuclei. It increased the binding of poly(A) to the envelope and increased the maximal catalytic rate of the NTPase, but it did not alter the apparent Km of the enzyme or the extent of its stimulation by poly(A). The nuclear envelope-associated protein kinase that down-regulates the NTPase was inhibited by the antibody, while other protein kinases were not affected. Because both the NTPase and mRNA efflux were inhibited by the tumor promoter, 12-O-tetradecanoylphorbol 13-acetate, the sensitive kinase is probably protein kinase C. Protein kinase C was found to be associated with the isolated nuclear envelope. The antibody reacted with both a Mr 83,000 and a Mr 65,000 nuclear envelope polypeptide from rat liver and other tissues. By immunofluorescence microscopy in CV-1 cells, the antibody localized to the nuclear envelope and, in addition, to cytoplasmic filaments which show some superposition with the microfilament network.

  14. Photoaffinity labeling of the major nucleosidetriphosphatase of rat liver nuclear envelope.

    PubMed

    Clawson, G A; Woo, C H; Button, J; Smuckler, E A

    1984-07-17

    We employed the photoaffinity probe 8-azido-adenosine 5'-triphosphate (aATP) to identify the nuclear envelope (NE) nucleosidetriphosphatase activity (NTPase) implicated in control of RNA transport. The photoprobe was hydrolyzed at rates comparable to those for ATP, with a Michaelis constant of 0.225 mM. Photolabeling was dependent upon UV irradiation (300-nm max) and was not affected by quercetin. Unlabeled ATP or GTP competed with [32P]aATP in photolabeling experiments, and UTP was a less effective competitor, paralleling the substrate specificity of the NTPase. Incubation of NE with aATP led to a UV, time, and concentration dependent irreversible inactivation of NTPase. The inactivation could be blocked by ATP or GTP. Polyacrylamide gel electrophoresis and autoradiography of photolabeled NE showed selective, UV-dependent labeling of a 46-kDa protein with both [gamma-32P]aATP and [alpha-32P]aATP. This band was not labeled with [gamma-32P]ATP. Since the NE NTPase implicated in RNA transport is modulated by RNA, we examined the effects of RNA on the labeling process. Removal of RNA from the NE preparations (by RNase/DNase digestion) reduced NTPase by 30-40% and eliminated photolabeling of the 46-kDa band. Addition of yeast RNA to such preparations increased NTPase activity to control levels and selectively reinstated photolabeling of the 46-kDa band. These results suggest that the 46-kDa protein represents the major NTPase implicated in RNA transport. PMID:6087895

  15. Nuclear envelope proteins modulate proliferation of vascular smooth muscle cells during cyclic stretch application.

    PubMed

    Qi, Ying-Xin; Yao, Qing-Ping; Huang, Kai; Shi, Qian; Zhang, Ping; Wang, Guo-Liang; Han, Yue; Bao, Han; Wang, Lu; Li, Hai-Peng; Shen, Bao-Rong; Wang, Yingxiao; Chien, Shu; Jiang, Zong-Lai

    2016-05-10

    Cyclic stretch is an important inducer of vascular smooth muscle cell (VSMC) proliferation, which is crucial in vascular remodeling during hypertension. However, the molecular mechanism remains unclear. We studied the effects of emerin and lamin A/C, two important nuclear envelope proteins, on VSMC proliferation in hypertension and the underlying mechano-mechanisms. In common carotid artery of hypertensive rats in vivo and in cultured cells subjected to high (15%) cyclic stretch in vitro, VSMC proliferation was increased significantly, and the expression of emerin and lamin A/C was repressed compared with normotensive or normal (5%) cyclic stretch controls. Using targeted siRNA to mimic the repressed expression of emerin or lamin A/C induced by 15% stretch, we found that VSMC proliferation was enhanced under static and 5%-stretch conditions. Overexpression of emerin or lamin A/C reversed VSMC proliferation induced by 15% stretch. Hence, emerin and lamin A/C play critical roles in suppressing VSMC hyperproliferation induced by hyperstretch. ChIP-on-chip and MOTIF analyses showed that the DNAs binding with emerin contain three transcription factor motifs: CCNGGA, CCMGCC, and ABTTCCG; DNAs binding with lamin A/C contain the motifs CVGGAA, GCCGCYGC, and DAAGAAA. Protein/DNA array proved that altered emerin or lamin A/C expression modulated the activation of various transcription factors. Furthermore, accelerating local expression of emerin or lamin A/C reversed cell proliferation in the carotid artery of hypertensive rats in vivo. Our findings establish the pathogenetic role of emerin and lamin A/C repression in stretch-induced VSMC proliferation and suggest mechanobiological mechanism underlying this process that involves the sequence-specific binding of emerin and lamin A/C to specific transcription factor motifs.

  16. Nuclear envelope proteins modulate proliferation of vascular smooth muscle cells during cyclic stretch application

    PubMed Central

    Qi, Ying-Xin; Yao, Qing-Ping; Huang, Kai; Shi, Qian; Zhang, Ping; Wang, Guo-Liang; Han, Yue; Bao, Han; Wang, Lu; Li, Hai-Peng; Shen, Bao-Rong; Wang, Yingxiao; Chien, Shu; Jiang, Zong-Lai

    2016-01-01

    Cyclic stretch is an important inducer of vascular smooth muscle cell (VSMC) proliferation, which is crucial in vascular remodeling during hypertension. However, the molecular mechanism remains unclear. We studied the effects of emerin and lamin A/C, two important nuclear envelope proteins, on VSMC proliferation in hypertension and the underlying mechano-mechanisms. In common carotid artery of hypertensive rats in vivo and in cultured cells subjected to high (15%) cyclic stretch in vitro, VSMC proliferation was increased significantly, and the expression of emerin and lamin A/C was repressed compared with normotensive or normal (5%) cyclic stretch controls. Using targeted siRNA to mimic the repressed expression of emerin or lamin A/C induced by 15% stretch, we found that VSMC proliferation was enhanced under static and 5%-stretch conditions. Overexpression of emerin or lamin A/C reversed VSMC proliferation induced by 15% stretch. Hence, emerin and lamin A/C play critical roles in suppressing VSMC hyperproliferation induced by hyperstretch. ChIP-on-chip and MOTIF analyses showed that the DNAs binding with emerin contain three transcription factor motifs: CCNGGA, CCMGCC, and ABTTCCG; DNAs binding with lamin A/C contain the motifs CVGGAA, GCCGCYGC, and DAAGAAA. Protein/DNA array proved that altered emerin or lamin A/C expression modulated the activation of various transcription factors. Furthermore, accelerating local expression of emerin or lamin A/C reversed cell proliferation in the carotid artery of hypertensive rats in vivo. Our findings establish the pathogenetic role of emerin and lamin A/C repression in stretch-induced VSMC proliferation and suggest mechanobiological mechanism underlying this process that involves the sequence-specific binding of emerin and lamin A/C to specific transcription factor motifs. PMID:27114541

  17. Quantified effects of chromosome-nuclear envelope attachments on 3D organization of chromosomes.

    PubMed

    Kinney, Nicholas Allen; Onufriev, Alexey V; Sharakhov, Igor V

    2015-01-01

    We use a combined experimental and computational approach to study the effects of chromosome-nuclear envelope (Chr-NE) attachments on the 3D genome organization of Drosophila melanogaster (fruit fly) salivary gland nuclei. We consider 3 distinct models: a Null model - without specific Chr-NE attachments, a 15-attachment model - with 15 previously known Chr-NE attachments, and a 48-attachment model - with 15 original and 33 recently identified Chr-NE attachments. The radial densities of chromosomes in the models are compared to the densities observed in 100 experimental images of optically sectioned salivary gland nuclei forming "z-stacks." Most of the experimental z-stacks support the Chr-NE 48-attachment model suggesting that as many as 48 chromosome loci with appreciable affinity for the NE are necessary to reproduce the experimentally observed distribution of chromosome density in fruit fly nuclei. Next, we investigate if and how the presence and the number of Chr-NE attachments affect several key characteristics of 3D genome organization: chromosome territories and gene-gene contacts. This analysis leads to novel insight about the possible role of Chr-NE attachments in regulating the genome architecture. Specifically, we find that model nuclei with more numerous Chr-NE attachments form more distinct chromosome territories and their chromosomes intertwine less frequently. Intra-chromosome and intra-arm contacts are more common in model nuclei with Chr-NE attachments compared to the Null model (no specific attachments), while inter-chromosome and inter-arm contacts are less common in nuclei with Chr-NE attachments. We demonstrate that Chr-NE attachments increase the specificity of long-range inter-chromosome and inter-arm contacts. The predicted effects of Chr-NE attachments are rationalized by intuitive volume vs. surface accessibility arguments.

  18. NUP145 encodes a novel yeast glycine-leucine-phenylalanine-glycine (GLFG) nucleoporin required for nuclear envelope structure

    PubMed Central

    1994-01-01

    We have isolated and characterized the gene encoding a fourth yeast glycine-leucine-phenylalanine-glycine (GLFG) repeat nucleoporin with a calculated molecular mass of 145.3 kD, and therefore termed NUP145. The amino-terminal half of Nup145p is similar to two previously identified GLFG nucleoporins, Nup116p and Nup100p (Wente, S. R., M. P. Rout, and G. Blobel. 1992. J. Cell Biol. 119:705-723). A deletion/disruption in the amino-terminal half of NUP145 (nup145 delta N) had only a slight effect on cell growth at temperatures between 17 and 37 degrees C. However, immunofluorescence microscopy of nup145 delta N cells with antinucleoporin antibodies showed that the characteristic punctate nuclear staining normally seen in wild-type yeast cells was reduced, with the majority of the signal located in one or two intense spots at the nuclear periphery. Thin section electron microscopy analysis revealed the presence of what appeared to be successive herniations of the nuclear envelope forming grape-like structures at primarily one site on the nup145 delta N nuclei. These successive herniations contained numerous NPC-like structures, correlating to the limited bright patches of anti-nucleoporin immunofluorescence signal. In some cases the successive herniations were small. Occasionally, however, multi-lobulated nuclei were seen. We suggest that the ultrastructural phenotype of nup145 delta N cells is due to a defective interaction of nup145 delta N NPCs with the surrounding pore membrane domain of the nuclear envelope. We have also analyzed the synthetic lethal phenotypes among GLFG nucleoporin mutant alleles, and found that strains harboring nup116 and either nup100 or nup145 mutations were not viable. This, in combination with the morphological analysis, may reflect overlapping yet distinct roles for these three GLFG nucleoporins in NPC-nuclear envelope interactions. PMID:8195299

  19. B-type nuclear lamin and the nuclear pore complex Nup107-160 influences maintenance of the spindle envelope required for cytokinesis in Drosophila male meiosis

    PubMed Central

    Hayashi, Daisuke; Tanabe, Karin; Katsube, Hiroka

    2016-01-01

    ABSTRACT In higher eukaryotes, nuclear envelope (NE) disassembly allows chromatin to condense and spindle microtubules to access kinetochores. The nuclear lamina, which strengthens the NE, is composed of a polymer meshwork made of A- and B-type lamins. We found that the B-type lamin (Lam) is not fully disassembled and continues to localize along the spindle envelope structure during Drosophila male meiosis I, while the A-type lamin (LamC) is completely dispersed throughout the cytoplasm. Among the nuclear pore complex proteins, Nup107 co-localized with Lam during this meiotic division. Surprisingly, Lam depletion resulted in a higher frequency of cytokinesis failure in male meiosis. We also observed the similar meiotic phenotype in Nup107-depleted cells. Abnormal localization of Lam was found in the Nup-depleted cells at premeiotic and meiotic stages. The central spindle microtubules became abnormal and recruitment of a contractile ring component to the cleavage sites was disrupted in Lam-depleted cells and Nup107-depleted cells. Therefore, we speculate that both proteins are required for a reinforcement of the spindle envelope, which supports the formation of central spindle microtubules essential for cytokinesis in Drosophila male meiosis. PMID:27402967

  20. B-type nuclear lamin and the nuclear pore complex Nup107-160 influences maintenance of the spindle envelope required for cytokinesis in Drosophila male meiosis.

    PubMed

    Hayashi, Daisuke; Tanabe, Karin; Katsube, Hiroka; Inoue, Yoshihiro H

    2016-08-15

    In higher eukaryotes, nuclear envelope (NE) disassembly allows chromatin to condense and spindle microtubules to access kinetochores. The nuclear lamina, which strengthens the NE, is composed of a polymer meshwork made of A- and B-type lamins. We found that the B-type lamin (Lam) is not fully disassembled and continues to localize along the spindle envelope structure during Drosophila male meiosis I, while the A-type lamin (LamC) is completely dispersed throughout the cytoplasm. Among the nuclear pore complex proteins, Nup107 co-localized with Lam during this meiotic division. Surprisingly, Lam depletion resulted in a higher frequency of cytokinesis failure in male meiosis. We also observed the similar meiotic phenotype in Nup107-depleted cells. Abnormal localization of Lam was found in the Nup-depleted cells at premeiotic and meiotic stages. The central spindle microtubules became abnormal and recruitment of a contractile ring component to the cleavage sites was disrupted in Lam-depleted cells and Nup107-depleted cells. Therefore, we speculate that both proteins are required for a reinforcement of the spindle envelope, which supports the formation of central spindle microtubules essential for cytokinesis in Drosophila male meiosis.

  1. GIP Contributions to the Regulation of Centromere at the Interface Between the Nuclear Envelope and the Nucleoplasm.

    PubMed

    Chabouté, Marie-Edith; Berr, Alexandre

    2016-01-01

    Centromeres are known as specific chromatin domains without which eukaryotic cells cannot divide properly during mitosis. Despite the considerable efforts to understand the centromere/kinetochore assembly during mitosis, until recently, comparatively few studies have dealt with the regulation of centromere during interphase. Here, we briefly review and discuss past and recent advances about the architecture of centromeres and their regulation during the cell cycle. Furthermore, we highlight and discuss new findings and hypotheses regarding the specific regulation of centromeres in both plant and animal nuclei, especially with GIP proteins at the interface between the nuclear envelope and the nucleoplasm. PMID:26904080

  2. GIP Contributions to the Regulation of Centromere at the Interface Between the Nuclear Envelope and the Nucleoplasm

    PubMed Central

    Chabouté, Marie-Edith; Berr, Alexandre

    2016-01-01

    Centromeres are known as specific chromatin domains without which eukaryotic cells cannot divide properly during mitosis. Despite the considerable efforts to understand the centromere/kinetochore assembly during mitosis, until recently, comparatively few studies have dealt with the regulation of centromere during interphase. Here, we briefly review and discuss past and recent advances about the architecture of centromeres and their regulation during the cell cycle. Furthermore, we highlight and discuss new findings and hypotheses regarding the specific regulation of centromeres in both plant and animal nuclei, especially with GIP proteins at the interface between the nuclear envelope and the nucleoplasm. PMID:26904080

  3. Herpes simplex virus glycoproteins gB and gH function in fusion between the virion envelope and the outer nuclear membrane.

    PubMed

    Farnsworth, Aaron; Wisner, Todd W; Webb, Michael; Roller, Richard; Cohen, Gary; Eisenberg, Roselyn; Johnson, David C

    2007-06-12

    Herpesviruses must traverse the nuclear envelope to gain access to the cytoplasm and, ultimately, to exit cells. It is believed that herpesvirus nucleocapsids enter the perinuclear space by budding through the inner nuclear membrane (NM). To reach the cytoplasm these enveloped particles must fuse with the outer NM and the unenveloped capsids then acquire a second envelope in the trans-Golgi network. Little is known about the process by which herpesviruses virions fuse with the outer NM. Here we show that a herpes simplex virus (HSV) mutant lacking both the two putative fusion glycoproteins gB and gH failed to cross the nuclear envelope. Enveloped virions accumulated in the perinuclear space or in membrane vesicles that bulged into the nucleoplasm (herniations). By contrast, mutants lacking just gB or gH showed only minor or no defects in nuclear egress. We concluded that either HSV gB or gH can promote fusion between the virion envelope and the outer NM. It is noteworthy that fusion associated with HSV entry requires the cooperative action of both gB and gH, suggesting that the two types of fusion (egress versus entry) are dissimilar processes.

  4. The BRCA1-binding protein BRAP2 can act as a cytoplasmic retention factor for nuclear and nuclear envelope-localizing testicular proteins.

    PubMed

    Davies, Rebecca G; Wagstaff, Kylie M; McLaughlin, Eileen A; Loveland, Kate L; Jans, David A

    2013-12-01

    Regulation of nuclear protein import is central to many cellular processes such as development, with a key mechanism being factors that retain cargoes in the cytoplasm that normally localize in the nucleus. The breast cancer antigen BRCA1-binding protein BRAP2 has been reported as a novel negative regulator of nuclear import of various nuclear localization signal (NLS)-containing viral and cellular proteins, but although implicated in differentiation pathways and highly expressed in tissues including testis, the gamut of targets for BRAP2 action in a developmental context is unknown. As a first step towards defining the BRAP2 interactome, we performed a yeast-2-hybrid screen to identify binding partners of BRAP2 in human testis. Here we report characterization for the first time of three of these: the high mobility group (HMG)-box-domain-containing chromatin component HMG20A, nuclear mitotic apparatus protein NuMA1 and synaptic nuclear envelope protein SYNE2. Co-immunoprecipitation experiments indicate association of BRAP2 with HMG20A, NuMA1, and SYNE2 in testis, underlining the physiological relevance of the interactions, with immunohistochemistry showing that where BRAP2 is co-expressed with HMG20A and NuMA1, both are present in the cytoplasm, in contrast to their nuclear localization in other testicular cell types. Importantly, quantitative confocal microscopic analysis of cultured cells indicates that ectopic expression of BRAP2 inhibits nuclear localization of HMG20A and NuMA1, and prevents nuclear envelope accumulation of SYNE2, the first report of BRAP2 altering localization of a non-nuclear protein. These results imply for the first time that BRAP2 may have an important role in modulating subcellular localization during testicular development. PMID:23707952

  5. Nuclear envelope dystrophies show a transcriptional fingerprint suggesting disruption of Rb-MyoD pathways in muscle regeneration.

    PubMed

    Bakay, Marina; Wang, Zuyi; Melcon, Gisela; Schiltz, Louis; Xuan, Jianhua; Zhao, Po; Sartorelli, Vittorio; Seo, Jinwook; Pegoraro, Elena; Angelini, Corrado; Shneiderman, Ben; Escolar, Diana; Chen, Yi-Wen; Winokur, Sara T; Pachman, Lauren M; Fan, Chenguang; Mandler, Raul; Nevo, Yoram; Gordon, Erynn; Zhu, Yitan; Dong, Yibin; Wang, Yue; Hoffman, Eric P

    2006-04-01

    Mutations of lamin A/C (LMNA) cause a wide range of human disorders, including progeria, lipodystrophy, neuropathies and autosomal dominant Emery-Dreifuss muscular dystrophy (EDMD). EDMD is also caused by X-linked recessive loss-of-function mutations of emerin, another component of the inner nuclear lamina that directly interacts with LMNA. One model for disease pathogenesis of LMNA and emerin mutations is cell-specific perturbations of the mRNA transcriptome in terminally differentiated cells. To test this model, we studied 125 human muscle biopsies from 13 diagnostic groups (125 U133A, 125 U133B microarrays), including EDMD patients with LMNA and emerin mutations. A Visual and Statistical Data Analyzer (VISDA) algorithm was used to statistically model cluster hierarchy, resulting in a tree of phenotypic classifications. Validations of the diagnostic tree included permutations of U133A and U133B arrays, and use of two probe set algorithms (MAS5.0 and MBEI). This showed that the two nuclear envelope defects (EDMD LMNA, EDMD emerin) were highly related disorders and were also related to fascioscapulohumeral muscular dystrophy (FSHD). FSHD has recently been hypothesized to involve abnormal interactions of chromatin with the nuclear envelope. To identify disease-specific transcripts for EDMD, we applied a leave-one-out (LOO) cross-validation approach using LMNA patient muscle as a test data set, with reverse transcription-polymerase chain reaction (RT-PCR) validations in both LMNA and emerin patient muscle. A high proportion of top-ranked and validated transcripts were components of the same transcriptional regulatory pathway involving Rb1 and MyoD during muscle regeneration (CRI-1, CREBBP, Nap1L1, ECREBBP/p300), where each was specifically upregulated in EDMD. Using a muscle regeneration time series (27 time points) we develop a transcriptional model for downstream consequences of LMNA and emerin mutations. We propose that key interactions between the nuclear

  6. The fission yeast NIMA kinase Fin1p is required for spindle function and nuclear envelope integrity

    PubMed Central

    Krien, Michael J.E.; West, Robert R.; John, Ulrik P.; Koniaras, Kalli; McIntosh, J.Richard; O’Connell, Matthew J.

    2002-01-01

    NIMA kinases appear to be the least functionally conserved mitotic regulators, being implicated in chromosome condensation in fungi and in spindle function in metazoans. We demonstrate here that the fission yeast NIMA homologue, Fin1p, can induce profound chromosome condensation in the absence of the condensin and topoisomerase II, indicating that Fin1p-induced condensation differs from mitotic condensation. Fin1p expression is transcriptionally and post-translationally cell cycle-regulated, with Fin1p kinase activity maximal from the metaphase–anaphase transition to G1. Fin1p is localized to the spindle pole body and fin1Δ cells are hypersensitive to anti-microtubule drugs, synthetically lethal with a number of spindle mutants and require the spindle checkpoint for viability. Moreover, fin1Δ cells show unusual and extensive elaborations of the nuclear envelope. These data support a role for Fin1p in spindle function and nuclear envelope transactions at or after the metaphase– anaphase transition that may be generally applicable to other NIMA-family members. PMID:11927555

  7. Growth of the nuclear envelope in the vegetative phase of the green alga Acetabularia. Evidence for assembly from membrane components synthesized in the cytoplasm

    PubMed Central

    1975-01-01

    The primary nucleus of the green alga Acetabularia grows about 25,000- fold in volume while it is separated from the endoplasmic reticulum and the whole cytoplasm by a special paranuclear cisterna of a vacuolar labyrinthum system which shows only very few (two to six per square micrometer) and small (ca. 40-120 nm in diamter) fenestrations. The nuclear envelope does not bear polyribosomes, nor do they occur in the entire zone intermediate between the nuclear envelope and the paranuclear cisterna. It is suggested that this special form of nuclear envelope growth takes place by assembly from cytoplasmically synthesized proteins that are translocated across the paranuclear cisterna in a nonmembrane-structured form. PMID:1158977

  8. Growth of the nuclear envelope in the vegetative phase of the green alga Acetabularia. Evidence for assembly from membrane components synthesized in the cytoplasm.

    PubMed

    Franke, W W; Spring, H; Scheer, U; Zerban, H

    1975-09-01

    The primary nucleus of the green alga Acetabularia grows about 25,000-fold in volume while it is separated from the endoplasmic reticulum and the whole cytoplasm by a special paranuclear cisterna of a vacuolar labyrinthum system which shows only very few (two to six per square micrometer) and small (ca. 40-120 nm in diamter) fenestrations. The nuclear envelope does not bear polyribosomes, nor do they occur in the entire zone intermediate between the nuclear envelope and the paranuclear cisterna. It is suggested that this special form of nuclear envelope growth takes place by assembly from cytoplasmically synthesized proteins that are translocated across the paranuclear cisterna in a nonmembrane-structured form.

  9. Structural protein 4.1R is integrally involved in nuclear envelope protein localization, centrosome–nucleus association and transcriptional signaling

    PubMed Central

    Meyer, Adam J.; Almendrala, Donna K.; Go, Minjoung M.; Krauss, Sharon Wald

    2011-01-01

    The multifunctional structural protein 4.1R is required for assembly and maintenance of functional nuclei but its nuclear roles are unidentified. 4.1R localizes within nuclei, at the nuclear envelope, and in cytoplasm. Here we show that 4.1R, the nuclear envelope protein emerin and the intermediate filament protein lamin A/C co-immunoprecipitate, and that 4.1R-specific depletion in human cells by RNA interference produces nuclear dysmorphology and selective mislocalization of proteins from several nuclear subcompartments. Such 4.1R-deficiency causes emerin to partially redistribute into the cytoplasm, whereas lamin A/C is disorganized at nuclear rims and displaced from nucleoplasmic foci. The nuclear envelope protein MAN1, nuclear pore proteins Tpr and Nup62, and nucleoplasmic proteins NuMA and LAP2α also have aberrant distributions, but lamin B and LAP2β have normal localizations. 4.1R-deficient mouse embryonic fibroblasts show a similar phenotype. We determined the functional effects of 4.1R-deficiency that reflect disruption of the association of 4.1R with emerin and A-type lamin: increased nucleus–centrosome distances, increased β-catenin signaling, and relocalization of β-catenin from the plasma membrane to the nucleus. Furthermore, emerin- and lamin-A/C-null cells have decreased nuclear 4.1R. Our data provide evidence that 4.1R has important functional interactions with emerin and A-type lamin that impact upon nuclear architecture, centrosome–nuclear envelope association and the regulation of β-catenin transcriptional co-activator activity that is dependent on β-catenin nuclear export. PMID:21486941

  10. A flow cytometry-based screen of nuclear envelope transmembrane proteins identifies NET4/Tmem53 as involved in stress-dependent cell cycle withdrawal.

    PubMed

    Korfali, Nadia; Srsen, Vlastimil; Waterfall, Martin; Batrakou, Dzmitry G; Pekovic, Vanja; Hutchison, Christopher J; Schirmer, Eric C

    2011-01-01

    Disruption of cell cycle regulation is one mechanism proposed for how nuclear envelope protein mutation can cause disease. Thus far only a few nuclear envelope proteins have been tested/found to affect cell cycle progression: to identify others, 39 novel nuclear envelope transmembrane proteins were screened for their ability to alter flow cytometry cell cycle/DNA content profiles when exogenously expressed. Eight had notable effects with seven increasing and one decreasing the 4N:2N ratio. We subsequently focused on NET4/Tmem53 that lost its effects in p53(-/-) cells and retinoblastoma protein-deficient cells. NET4/TMEM53 knockdown by siRNA altered flow cytometry cell cycle/DNA content profiles in a similar way as overexpression. NET4/TMEM53 knockdown did not affect total retinoblastoma protein levels, unlike nuclear envelope-associated proteins Lamin A and LAP2α. However, a decrease in phosphorylated retinoblastoma protein was observed along with a doubling of p53 levels and a 7-fold increase in p21. Consequently cells withdrew from the cell cycle, which was confirmed in MRC5 cells by a drop in the percentage of cells expressing Ki-67 antigen and an increase in the number of cells stained for ß-galactosidase. The ß-galactosidase upregulation suggests that cells become prematurely senescent. Finally, the changes in retinoblastoma protein, p53, and p21 resulting from loss of NET4/Tmem53 were dependent upon active p38 MAP kinase. The finding that roughly a fifth of nuclear envelope transmembrane proteins screened yielded alterations in flow cytometry cell cycle/DNA content profiles suggests a much greater influence of the nuclear envelope on the cell cycle than is widely held.

  11. Investigation of the Chromosome Regions with Significant Affinity for the Nuclear Envelope in Fruit Fly – A Model Based Approach

    PubMed Central

    Kinney, Nicholas Allen; Sharakhov, Igor V.; Onufriev, Alexey V.

    2014-01-01

    Three dimensional nuclear architecture is important for genome function, but is still poorly understood. In particular, little is known about the role of the “boundary conditions” – points of attachment between chromosomes and the nuclear envelope. We describe a method for modeling the 3D organization of the interphase nucleus, and its application to analysis of chromosome-nuclear envelope (Chr-NE) attachments of polytene (giant) chromosomes in Drosophila melanogaster salivary glands. The model represents chromosomes as self-avoiding polymer chains confined within the nucleus; parameters of the model are taken directly from experiment, no fitting parameters are introduced. Methods are developed to objectively quantify chromosome territories and intertwining, which are discussed in the context of corresponding experimental observations. In particular, a mathematically rigorous definition of a territory based on convex hull is proposed. The self-avoiding polymer model is used to re-analyze previous experimental data; the analysis suggests 33 additional Chr-NE attachments in addition to the 15 already explored Chr-NE attachments. Most of these new Chr-NE attachments correspond to intercalary heterochromatin – gene poor, dark staining, late replicating regions of the genome; however, three correspond to euchromatin – gene rich, light staining, early replicating regions of the genome. The analysis also suggests 5 regions of anti-contact, characterized by aversion for the NE, only two of these correspond to euchromatin. This composition of chromatin suggests that heterochromatin may not be necessary or sufficient for the formation of a Chr-NE attachment. To the extent that the proposed model represents reality, the confinement of the polytene chromosomes in a spherical nucleus alone does not favor the positioning of specific chromosome regions at the NE as seen in experiment; consequently, the 15 experimentally known Chr-NE attachment positions do not appear to

  12. Human Cytomegalovirus nuclear egress and secondary envelopment are negatively affected in the absence of cellular p53.

    PubMed

    Kuan, Man I; O'Dowd, John M; Chughtai, Kamila; Hayman, Ian; Brown, Celeste J; Fortunato, Elizabeth A

    2016-10-01

    Human Cytomegalovirus (HCMV) infection is compromised in cells lacking p53, a transcription factor that mediates cellular stress responses. In this study we have investigated compromised functional virion production in cells with p53 knocked out (p53KOs). Infectious center assays found most p53KOs released functional virions. Analysis of electron micrographs revealed modestly decreased capsid production in infected p53KOs compared to wt. Substantially fewer p53KOs displayed HCMV-induced infoldings of the inner nuclear membrane (IINMs). In p53KOs, fewer capsids were found in IINMs and in the cytoplasm. The deficit in virus-induced membrane remodeling within the nucleus of p53KOs was mirrored in the cytoplasm, with a disproportionately smaller number of capsids re-enveloped. Reintroduction of p53 substantially recovered these deficits. Overall, the absence of p53 contributed to inhibition of the formation and function of IINMs and re-envelopment of the reduced number of capsids able to reach the cytoplasm.

  13. Herpesvirus gB-induced fusion between the virion envelope and outer nuclear membrane during virus egress is regulated by the viral US3 kinase.

    PubMed

    Wisner, Todd W; Wright, Catherine C; Kato, Akihisa; Kawaguchi, Yasushi; Mou, Fan; Baines, Joel D; Roller, Richard J; Johnson, David C

    2009-04-01

    Herpesvirus capsids collect along the inner surface of the nuclear envelope and bud into the perinuclear space. Enveloped virions then fuse with the outer nuclear membrane (NM). We previously showed that herpes simplex virus (HSV) glycoproteins gB and gH act in a redundant fashion to promote fusion between the virion envelope and the outer NM. HSV mutants lacking both gB and gH accumulate enveloped virions in herniations, vesicles that bulge into the nucleoplasm. Earlier studies had shown that HSV mutants lacking the viral serine/threonine kinase US3 also accumulate herniations. Here, we demonstrate that HSV gB is phosphorylated in a US3-dependent manner in HSV-infected cells, especially in a crude nuclear fraction. Moreover, US3 directly phosphorylated the gB cytoplasmic (CT) domain in in vitro assays. Deletion of gB in the context of a US3-null virus did not add substantially to defects in nuclear egress. The majority of the US3-dependent phosphorylation of gB involved the CT domain and amino acid T887, a residue present in a motif similar to that recognized by US3 in other proteins. HSV recombinants lacking gH and expressing either gB substitution mutation T887A or a gB truncated at residue 886 displayed substantial defects in nuclear egress. We concluded that phosphorylation of the gB CT domain is important for gB-mediated fusion with the outer NM. This suggested a model in which the US3 kinase is incorporated into the tegument layer (between the capsid and envelope) in HSV virions present in the perinuclear space. By this packaging, US3 might be brought close to the gB CT tail, leading to phosphorylation and triggering fusion between the virion envelope and the outer NM.

  14. [Comparative Analysis of DNA Sequences of Regions of X-Chromosome Attachment to the Nuclear Envelope of Nurse Cells Anopheles messeae Fall].

    PubMed

    Artemov, G N; Vasil'eva, O Yu; Stegniy, V N

    2015-07-01

    Polytene chromosomes of ovarian nurse cells of Anopheles mosquitoes form strong contacts with the nuclear envelope. The presence of contacts, their position at nurse cell chromosomes, and their morphological features are species-specific in malaria mosquitoes. It is important to determine the nature of these interspecies differences in the nuclear architecture, both to understand the function of the nucleus and to assess the role of the spatial organization of chromosomes in evolution. Using dot-blot hybridization, we compared DNA sequences of the clone library from the X-chromosome attachment region to the nuclear envelope of ovarian nurse cells of Anopheles messeae with DNA-probes: (1) of the X-chromosome attachment region of An. atroparvus, (2) of the 3R chromosome attachment region ofAn. messeae, and (3) of the chromosome 2 pericentromeric region of An. messeae, without expressed contacts with the nuclear envelope. It has been shown that the chromosome attachment regions have a significantly higher number of homologous DNA sequences as compared with the pericentromeric region of chromosome 2. Sequences that are common for attachment regions are largely potentially able to participate in the formation of chromatin loop domains and to interact with some nucleus frameworks, according to the analysis in the ChrClass program. The obtained results support the important role of DNA in the formation of strong chromosomal attachments to the nuclear envelope in nurse cells of Anopheles mosquitoes.

  15. Meiotic chromosomes move by linkage to dynamic actin cables with transduction of force through the nuclear envelope.

    PubMed

    Koszul, R; Kim, K P; Prentiss, M; Kleckner, N; Kameoka, S

    2008-06-27

    Chromosome movement is prominent during meiosis. Here, using a combination of in vitro and in vivo approaches, we elucidate the basis for dynamic mid-prophase telomere-led chromosome motion in budding yeast. Diverse findings reveal a process in which, at the pachytene stage, individual telomere/nuclear envelope (NE) ensembles attach passively to, and then move in concert with, nucleus-hugging actin cables that are continuous with the global cytoskeletal actin network. Other chromosomes move in concert with lead chromosome(s). The same process, in modulated form, explains the zygotene "bouquet" configuration in which, immediately preceding pachytene, chromosome ends colocalize dynamically in a restricted region of the NE. Mechanical properties of the system and biological roles of mid-prophase movement for meiosis, including recombination, are discussed. PMID:18585353

  16. Light up Live Cell Nuclear Envelope in Real-Time Using a Two-Photon Absorption and AIE Chromophore.

    PubMed

    Tian, Xiaohe; Wang, Hui; Guan, Lijuan; Zhang, Qiong; Zhou, Hongping; Li, Chunxia; Huang, Bei; Wu, Jieying; Tian, Yupeng

    2016-01-01

    In this letter, aggregation from two-photon absorption (2PA) molecules in living cells were firstly observed and the related aggregation induced emission (AIE) properties were investigated as a cell tracer for L ((Z)-3-(4-(Bis(4-ethoxyphenyl) amino)phenyl)-2-(4-amino-phenyl)- acrylonitrile cyano-substituted ) based on triphenylamine with D-π-A model. L was further used as a two-photon absorption (2PA, λex = 900, λem = 550 nm δ = 156 GM) live-cell marker for real-time, long-term cell growth and proliferation monitoring, with rapidly adhering whole intracellular membrane-rich system. Remarkably, different from existing organic AIE chromophores and other commercially available probes, L exhibited intense intracellular-AIE property with stable nuclear envelope (NE) staining under two-photon excited microscopy (TPEM) through detailed in cellulo studies.

  17. Inhibition of the host translation shutoff response by herpes simplex virus 1 triggers nuclear envelope-derived autophagy.

    PubMed

    Radtke, Kerstin; English, Luc; Rondeau, Christiane; Leib, David; Lippé, Roger; Desjardins, Michel

    2013-04-01

    Macroautophagy is a cellular pathway that degrades intracellular pathogens and contributes to antigen presentation. Herpes simplex virus 1 (HSV-1) infection triggers both macroautophagy and an additional form of autophagy that uses the nuclear envelope as a source of membrane. The present study constitutes the first in-depth analysis of nuclear envelope-derived autophagy (NEDA). We established LC3a as a marker that allowed us to distinguish between NEDA and macroautophagy in both immunofluorescence and flow cytometry. NEDA was observed in many different cell types, indicating that it is a general response to HSV-1 infection. This autophagic pathway is known to depend on the viral protein γ34.5, which can inhibit macroautophagy via binding to beclin-1. Using mutant viruses, we were able to show that binding of beclin-1 by γ34.5 had no effect on NEDA, demonstrating that NEDA is regulated differently than macroautophagy. Instead, NEDA was triggered in response to γ34.5 binding to protein phosphatase 1α, an interaction used by the virus to prevent host cells from shutting off protein translation. NEDA was not triggered when late viral protein production was inhibited with acyclovir or hippuristanol, indicating that the accumulation of these proteins might stress infected cells. Interestingly, expression of the late viral protein gH was sufficient to rescue NEDA in the context of infection with a virus that otherwise does not support strong late viral protein expression. We argue that NEDA is a cellular stress response triggered late during HSV-1 infection and might compensate for the viral alteration of the macroautophagic response.

  18. Do nuclear envelope and intranuclear proteins reorganize during mitosis to form an elastic, hydrogel-like spindle matrix?

    PubMed

    Johansen, Kristen M; Forer, Arthur; Yao, Changfu; Girton, Jack; Johansen, Jørgen

    2011-04-01

    The idea of a spindle matrix has long been proposed in order to account for poorly understood features of mitosis. However, its molecular nature and structural composition have remained elusive. Here, we propose that the spindle matrix may be constituted by mainly nuclear-derived proteins that reorganize during the cell cycle to form an elastic gel-like matrix. We discuss this hypothesis in the context of recent observations from phylogenetically diverse organisms that nuclear envelope and intranuclear proteins form a highly dynamic and malleable structure that contributes to mitotic spindle function. We suggest that the viscoelastic properties of such a matrix may constrain spindle length while at the same time facilitating microtubule growth and dynamics as well as chromosome movement. A corollary to this hypothesis is that a key determinant of spindle size may be the amount of nuclear proteins available to form the spindle matrix. Such a matrix could also serve as a spatial regulator of spindle assembly checkpoint proteins during open and semi-open mitosis. PMID:21274615

  19. Nuclear Envelope Composition Determines the Ability of Neutrophil-type Cells to Passage through Micron-scale Constrictions*

    PubMed Central

    Rowat, Amy C.; Jaalouk, Diana E.; Zwerger, Monika; Ung, W. Lloyd; Eydelnant, Irwin A.; Olins, Don E.; Olins, Ada L.; Herrmann, Harald; Weitz, David A.; Lammerding, Jan

    2013-01-01

    Neutrophils are characterized by their distinct nuclear shape, which is thought to facilitate the transit of these cells through pore spaces less than one-fifth of their diameter. We used human promyelocytic leukemia (HL-60) cells as a model system to investigate the effect of nuclear shape in whole cell deformability. We probed neutrophil-differentiated HL-60 cells lacking expression of lamin B receptor, which fail to develop lobulated nuclei during granulopoiesis and present an in vitro model for Pelger-Huët anomaly; despite the circular morphology of their nuclei, the cells passed through micron-scale constrictions on similar timescales as scrambled controls. We then investigated the unique nuclear envelope composition of neutrophil-differentiated HL-60 cells, which may also impact their deformability; although lamin A is typically down-regulated during granulopoiesis, we genetically modified HL-60 cells to generate a subpopulation of cells with well defined levels of ectopic lamin A. The lamin A-overexpressing neutrophil-type cells showed similar functional characteristics as the mock controls, but they had an impaired ability to pass through micron-scale constrictions. Our results suggest that levels of lamin A have a marked effect on the ability of neutrophils to passage through micron-scale constrictions, whereas the unusual multilobed shape of the neutrophil nucleus is less essential. PMID:23355469

  20. NSF- and SNARE-mediated membrane fusion is required for nuclear envelope formation and completion of nuclear pore complex assembly in Xenopus laevis egg extracts.

    PubMed

    Baur, Tina; Ramadan, Kristijan; Schlundt, Andreas; Kartenbeck, Jürgen; Meyer, Hemmo H

    2007-08-15

    Despite the progress in understanding nuclear envelope (NE) reformation after mitosis, it has remained unclear what drives the required membrane fusion and how exactly this is coordinated with nuclear pore complex (NPC) assembly. Here, we show that, like other intracellular fusion reactions, NE fusion in Xenopus laevis egg extracts is mediated by SNARE proteins that require activation by NSF. Antibodies against Xenopus NSF, depletion of NSF or the dominant-negative NSF(E329Q) variant specifically inhibited NE formation. Staging experiments further revealed that NSF was required until sealing of the envelope was completed. Moreover, excess exogenous alpha-SNAP that blocks SNARE function prevented membrane fusion and caused accumulation of non-flattened vesicles on the chromatin surface. Under these conditions, the nucleoporins Nup107 and gp210 were fully recruited, whereas assembly of FxFG-repeat-containing nucleoporins was blocked. Together, we define NSF- and SNARE-mediated membrane fusion events as essential steps during NE formation downstream of Nup107 recruitment, and upstream of membrane flattening and completion of NPC assembly.

  1. SAFEGUARDS ENVELOPE

    SciTech Connect

    Duc Cao; Richard Metcalf

    2010-07-01

    The Safeguards Envelope is a strategy to determine a set of specific operating parameters within which nuclear facilities may operate to maximize safeguards effectiveness without sacrificing safety or plant efficiency. This paper details advanced statistical techniques that will be applied to real plant process monitoring (PM) data from the Idaho Chemical Processing Plant (ICPP). In a simulation based on this data, multi-tank and multi-attribute correlations were tested against synthetic diversion scenarios. Kernel regression smoothing was used to fit a curve to the historical data, and multivariable, residual analysis and cumulative sum techniques set parameters for operating conditions. Diversion scenarios were created and tested, showing improved results when compared with a previous study utilizing only one-variable Z-testing. A brief analysis of the impact of the safeguards optimization on the rest of plant efficiency, criticality concerns, and overall requirements is presented.

  2. TorsinA binds the KASH domain of nesprins and participates in linkage between nuclear envelope and cytoskeleton.

    PubMed

    Nery, Flávia C; Zeng, Juan; Niland, Brian P; Hewett, Jeffrey; Farley, Jonathan; Irimia, Daniel; Li, Yuqing; Wiche, Gerhard; Sonnenberg, Arnoud; Breakefield, Xandra O

    2008-10-15

    A specific mutation (DeltaE) in torsinA underlies most cases of the dominantly inherited movement disorder, early-onset torsion dystonia (DYT1). TorsinA, a member of the AAA+ ATPase superfamily, is located within the lumen of the nuclear envelope (NE) and endoplasmic reticulum (ER). We investigated an association between torsinA and nesprin-3, which spans the outer nuclear membrane (ONM) of the NE and links it to vimentin via plectin in fibroblasts. Mouse nesprin-3alpha co-immunoprecipitated with torsinA and this involved the C-terminal region of torsinA and the KASH domain of nesprin-3alpha. This association with human nesprin-3 appeared to be stronger for torsinADeltaE than for torsinA. TorsinA also associated with the KASH domains of nesprin-1 and -2 (SYNE1 and 2), which link to actin. In the absence of torsinA, in knockout mouse embryonic fibroblasts (MEFs), nesprin-3alpha was localized predominantly in the ER. Enrichment of yellow fluorescent protein (YFP)-nesprin-3 in the ER was also seen in the fibroblasts of DYT1 patients, with formation of YFP-positive globular structures enriched in torsinA, vimentin and actin. TorsinA-null MEFs had normal NE structure, but nuclear polarization and cell migration were delayed in a wound-healing assay, as compared with wild-type MEFs. These studies support a role for torsinA in dynamic interactions between the KASH domains of nesprins and their protein partners in the lumen of the NE, with torsinA influencing the localization of nesprins and associated cytoskeletal elements and affecting their role in nuclear and cell movement.

  3. The SUN protein UNC-84 is required only in force-bearing cells to maintain nuclear envelope architecture.

    PubMed

    Cain, Natalie E; Tapley, Erin C; McDonald, Kent L; Cain, Benjamin M; Starr, Daniel A

    2014-07-21

    The nuclear envelope (NE) consists of two evenly spaced bilayers, the inner and outer nuclear membranes. The Sad1p and UNC-84 (SUN) proteins and Klarsicht, ANC-1, and Syne homology (KASH) proteins that interact to form LINC (linker of nucleoskeleton and cytoskeleton) complexes connecting the nucleoskeleton to the cytoskeleton have been implicated in maintaining NE spacing. Surprisingly, the NE morphology of most Caenorhabditis elegans nuclei was normal in the absence of functional SUN proteins. Distortions of the perinuclear space observed in unc-84 mutant muscle nuclei resembled those previously observed in HeLa cells, suggesting that SUN proteins are required to maintain NE architecture in cells under high mechanical strain. The UNC-84 protein with large deletions in its luminal domain was able to form functional NE bridges but had no observable effect on NE architecture. Therefore, SUN-KASH bridges are only required to maintain NE spacing in cells subjected to increased mechanical forces. Furthermore, SUN proteins do not dictate the width of the NE.

  4. MCLIP, an effective method to detect interactions of transmembrane proteins of the nuclear envelope in live cells.

    PubMed

    Jafferali, Mohammed Hakim; Vijayaraghavan, Balaje; Figueroa, Ricardo A; Crafoord, Ellinor; Gudise, Santhosh; Larsson, Veronica J; Hallberg, Einar

    2014-10-01

    Investigating interactions of proteins in the nuclear envelope (NE) using co-immunoprecipitation (Co-IP) has previously been difficult or even impossible due to their inherent resistance to extraction. We have developed a novel method, MCLIP (Membrane protein Cross-Link ImmunoPrecipitation), which takes advantage of a cell permeable crosslinker to enable effective detection and analysis of specific interactions of NE proteins in live cells using Western blot. Using MCLIP we show that, in U2OS cells, the integral inner nuclear membrane protein Samp1 interacts with Lamin B1, the LINC (Linker of nucleoskeleton and cytoskeleton) complex protein, Sun1 and the soluble small GTPase Ran. The results show that the previously detected in vitro interaction between Samp1 and Emerin also takes place in live cells. In vitro pull down experiments show, that the nucleoplasmic domains of Samp1 and Emerin can bind directly to each other. We also, show that MCLIP is suitable to coprecipitate protein interactions in different stages of the cell cycle.

  5. HA95 and LAP2 beta mediate a novel chromatin-nuclear envelope interaction implicated in initiation of DNA replication.

    PubMed

    Martins, Sandra; Eikvar, Sissel; Furukawa, Kazuhiro; Collas, Philippe

    2003-01-20

    HA95 is a chromatin-associated protein that interfaces the nuclear envelope (NE) and chromatin. We report an interaction between HA95 and the inner nuclear membrane protein lamina-associated polypeptide (LAP) 2 beta, and a role of this association in initiation of DNA replication. Precipitation of GST-LAP2 beta fusion proteins and overlays of immobilized HA95 indicate that a first HA95-binding region lies within amino acids 137-242 of LAP2 beta. A second domain sufficient to bind HA95 colocalizes with the lamin B-binding domain of LAP2beta at residues 299-373. HA95-LAP2 beta interaction is not required for NE formation. However, disruption of the association of HA95 with the NH2-terminal HA95-binding domain of LAP2 beta abolishes the initiation, but not elongation, of DNA replication in purified G1 phase nuclei incubated in S-phase extract. Inhibition of replication initiation correlates with proteasome-mediated proteolysis of Cdc6, a component of the prereplication complex. Rescue of Cdc6 degradation with proteasome inhibitors restores replication. We propose that an interaction of LAP2beta, or LAP2 proteins, with HA95 is involved in the control of initiation of DNA replication. PMID:12538639

  6. Nuclear envelope alterations generate an aging-like epigenetic pattern in mice deficient in Zmpste24 metalloprotease.

    PubMed

    Osorio, Fernando G; Varela, Ignacio; Lara, Ester; Puente, Xose S; Espada, Jesús; Santoro, Raffaella; Freije, José M P; Fraga, Mario F; López-Otín, Carlos

    2010-12-01

    Mutations in the nuclear envelope protein lamin A or in its processing protease ZMPSTE24 cause human accelerated aging syndromes, including Hutchinson-Gilford progeria syndrome. Similarly, Zmpste24-deficient mice accumulate unprocessed prelamin A and develop multiple progeroid symptoms, thus representing a valuable animal model for the study of these syndromes. Zmpste24-deficient mice also show marked transcriptional alterations associated with chromatin disorganization, but the molecular links between both processes are unknown. We report herein that Zmpste24-deficient mice show a hypermethylation of rDNA that reduces the transcription of ribosomal genes, being this reduction reversible upon treatment with DNA methyltransferase inhibitors. This alteration has been previously described during physiological aging in rodents, suggesting its potential role in the development of the progeroid phenotypes. We also show that Zmpste24-deficient mice present global hypoacetylation of histones H2B and H4. By using a combination of RNA sequencing and chromatin immunoprecipitation assays, we demonstrate that these histone modifications are associated with changes in the expression of several genes involved in the control of cell proliferation and metabolic processes, which may contribute to the plethora of progeroid symptoms exhibited by Zmpste24-deficient mice. The identification of these altered genes may help to clarify the molecular mechanisms underlying aging and progeroid syndromes as well as to define new targets for the treatment of these dramatic diseases.

  7. Essential role of the Cdk2 activator RingoA in meiotic telomere tethering to the nuclear envelope

    PubMed Central

    Mikolcevic, Petra; Isoda, Michitaka; Shibuya, Hiroki; del Barco Barrantes, Ivan; Igea, Ana; Suja, José A.; Shackleton, Sue; Watanabe, Yoshinori; Nebreda, Angel R.

    2016-01-01

    Cyclin-dependent kinases (CDKs) play key roles in cell cycle regulation. Genetic analysis in mice has revealed an essential role for Cdk2 in meiosis, which renders Cdk2 knockout (KO) mice sterile. Here we show that mice deficient in RingoA, an atypical activator of Cdk1 and Cdk2 that has no amino acid sequence homology to cyclins, are sterile and display meiotic defects virtually identical to those observed in Cdk2 KO mice including non-homologous chromosome pairing, unrepaired double-strand breaks, undetectable sex-body and pachytene arrest. Interestingly, RingoA is required for Cdk2 targeting to telomeres and RingoA KO spermatocytes display severely affected telomere tethering as well as impaired distribution of Sun1, a protein essential for the attachment of telomeres to the nuclear envelope. Our results identify RingoA as an important activator of Cdk2 at meiotic telomeres, and provide genetic evidence for a physiological function of mammalian Cdk2 that is not dependent on cyclins. PMID:27025256

  8. Kar1 binding to Sfi1 C-terminal regions anchors the SPB bridge to the nuclear envelope

    PubMed Central

    Seybold, Christian; Elserafy, Menattallah; Rüthnick, Diana; Ozboyaci, Musa; Neuner, Annett; Flottmann, Benjamin; Heilemann, Mike; Wade, Rebecca C.

    2015-01-01

    The yeast spindle pole body (SPB) is the functional equivalent of the mammalian centrosome. The half bridge is a SPB substructure on the nuclear envelope (NE), playing a key role in SPB duplication. Its cytoplasmic components are the membrane-anchored Kar1, the yeast centrin Cdc31, and the Cdc31-binding protein Sfi1. In G1, the half bridge expands into the bridge through Sfi1 C-terminal (Sfi1-CT) dimerization, the licensing step for SPB duplication. We exploited photo-activated localization microscopy (PALM) to show that Kar1 localizes in the bridge center. Binding assays revealed direct interaction between Kar1 and C-terminal Sfi1 fragments. kar1Δ cells whose viability was maintained by the dominant CDC31-16 showed an arched bridge, indicating Kar1’s function in tethering Sfi1 to the NE. Cdc31-16 enhanced Cdc31–Cdc31 interactions between Sfi1–Cdc31 layers, as suggested by binding free energy calculations. In our model, Kar1 binding is restricted to Sfi1-CT and Sfi1 C-terminal centrin-binding repeats, and centrin and Kar1 provide cross-links, while Sfi1-CT stabilizes the bridge and ensures timely SPB separation. PMID:26076691

  9. Saccharomyces cerevisiae MPS2 Encodes a Membrane Protein Localized at the Spindle Pole Body and the Nuclear Envelope

    PubMed Central

    Muñoz-Centeno, María de la Cruz; McBratney, Susan; Monterrosa, Antonio; Byers, Breck; Mann, Carl; Winey, Mark

    1999-01-01

    The MPS2 (monopolar spindle two) gene is one of several genes required for the proper execution of spindle pole body (SPB) duplication in the budding yeast Saccharomyces cerevisiae (Winey et al., 1991). We report here that the MPS2 gene encodes an essential 44-kDa protein with two putative coiled-coil regions and a hydrophobic sequence. Although MPS2 is required for normal mitotic growth, some null strains can survive; these survivors exhibit slow growth and abnormal ploidy. The MPS2 protein was tagged with nine copies of the myc epitope, and biochemical fractionation experiments show that it is an integral membrane protein. Visualization of a green fluorescent protein (GFP) Mps2p fusion protein in living cells and indirect immunofluorescence microscopy of 9xmyc-Mps2p revealed a perinuclear localization with one or two brighter foci of staining corresponding to the SPB. Additionally, immunoelectron microscopy shows that GFP-Mps2p localizes to the SPB. Our analysis suggests that Mps2p is required as a component of the SPB for insertion of the nascent SPB into the nuclear envelope. PMID:10397772

  10. SUN anchors pollen WIP–WIT complexes at the vegetative nuclear envelope and is necessary for pollen tube targeting and fertility

    PubMed Central

    Zhou, Xiao; Groves, Norman Reid; Meier, Iris

    2015-01-01

    LINC (linker of nucleoskeleton and cytoskeleton) complexes play an essential role in nuclear migration by connecting the nucleus to the cytoskeleton and/or motor proteins. Plant LINC complexes have recently been identified in Arabidopsis thaliana, with the inner nuclear membrane SUN and outer nuclear membrane WIP proteins comprising the first identified complex. A recent study identified a nuclear movement defect in Arabidopsis pollen vegetative nuclei linked to the outer nuclear envelope WIP and WIT proteins. However, the role that SUN proteins may play in pollen nuclear migration has yet to be addressed. To explore this question, a SUN2 lumenal domain that was targeted to the ER specifically in pollen was over-expressed. It is shown that the ER-targeted SUN2 lumenal domain was able to displace WIP and WIT proteins from the pollen vegetative nuclear envelope. Expression of this dominant-negative transgene led to impaired VN mobility, impaired pollen tube guidance, and defective pollen tube reception. The observed pollen defects are similar to phenotypes observed in a wip1-1 wip2-1 wip3-1 wit1-1 wit2-1 mutant. It is also shown that these defects were dependent on the KASH-binding function of the SUN2 lumenal domain. These data support a model where LINC complexes formed by SUN, WIP, and WIT at the VNE are responsible for VN migration and suggest an important function of SUN, WIP, and WIT in pollen tube guidance and reception. PMID:26409047

  11. SUN anchors pollen WIP-WIT complexes at the vegetative nuclear envelope and is necessary for pollen tube targeting and fertility.

    PubMed

    Zhou, Xiao; Groves, Norman Reid; Meier, Iris

    2015-12-01

    LINC (linker of nucleoskeleton and cytoskeleton) complexes play an essential role in nuclear migration by connecting the nucleus to the cytoskeleton and/or motor proteins. Plant LINC complexes have recently been identified in Arabidopsis thaliana, with the inner nuclear membrane SUN and outer nuclear membrane WIP proteins comprising the first identified complex. A recent study identified a nuclear movement defect in Arabidopsis pollen vegetative nuclei linked to the outer nuclear envelope WIP and WIT proteins. However, the role that SUN proteins may play in pollen nuclear migration has yet to be addressed. To explore this question, a SUN2 lumenal domain that was targeted to the ER specifically in pollen was over-expressed. It is shown that the ER-targeted SUN2 lumenal domain was able to displace WIP and WIT proteins from the pollen vegetative nuclear envelope. Expression of this dominant-negative transgene led to impaired VN mobility, impaired pollen tube guidance, and defective pollen tube reception. The observed pollen defects are similar to phenotypes observed in a wip1-1 wip2-1 wip3-1 wit1-1 wit2-1 mutant. It is also shown that these defects were dependent on the KASH-binding function of the SUN2 lumenal domain. These data support a model where LINC complexes formed by SUN, WIP, and WIT at the VNE are responsible for VN migration and suggest an important function of SUN, WIP, and WIT in pollen tube guidance and reception.

  12. Sigma-1 receptor mediates cocaine-induced transcriptional regulation by recruiting chromatin-remodeling factors at the nuclear envelope.

    PubMed

    Tsai, Shang-Yi A; Chuang, Jian-Ying; Tsai, Meng-Shan; Wang, Xiao-Fei; Xi, Zheng-Xiong; Hung, Jan-Jong; Chang, Wen-Chang; Bonci, Antonello; Su, Tsung-Ping

    2015-11-24

    The sigma-1 receptor (Sig-1R) chaperone at the endoplasmic reticulum (ER) plays important roles in cellular regulation. Here we found a new function of Sig-1R, in that it translocates from the ER to the nuclear envelope (NE) to recruit chromatin-remodeling molecules and regulate the gene transcription thereof. Sig-1Rs mainly reside at the ER-mitochondrion interface. However, on stimulation by agonists such as cocaine, Sig-1Rs translocate from ER to the NE, where Sig-1Rs bind NE protein emerin and recruit chromatin-remodeling molecules, including lamin A/C, barrier-to-autointegration factor (BAF), and histone deacetylase (HDAC), to form a complex with the gene repressor specific protein 3 (Sp3). Knockdown of Sig-1Rs attenuates the complex formation. Cocaine was found to suppress the gene expression of monoamine oxidase B (MAOB) in the brain of wild-type but not Sig-1R knockout mouse. A single dose of cocaine (20 mg/kg) in rats suppresses the level of MAOB at nuclear accumbens without affecting the level of dopamine transporter. Daily injections of cocaine in rats caused behavioral sensitization. Withdrawal from cocaine in cocaine-sensitized rats induced an apparent time-dependent rebound of the MAOB protein level to about 200% over control on day 14 after withdrawal. Treatment of cocaine-withdrawn rats with the MAOB inhibitor deprenyl completely alleviated the behavioral sensitization to cocaine. Our results demonstrate a role of Sig-1R in transcriptional regulation and suggest cocaine may work through this newly discovered genomic action to achieve its addictive action. Results also suggest the MAOB inhibitor deprenyl as a therapeutic agent to block certain actions of cocaine during withdrawal. PMID:26554014

  13. Sigma-1 receptor mediates cocaine-induced transcriptional regulation by recruiting chromatin-remodeling factors at the nuclear envelope.

    PubMed

    Tsai, Shang-Yi A; Chuang, Jian-Ying; Tsai, Meng-Shan; Wang, Xiao-Fei; Xi, Zheng-Xiong; Hung, Jan-Jong; Chang, Wen-Chang; Bonci, Antonello; Su, Tsung-Ping

    2015-11-24

    The sigma-1 receptor (Sig-1R) chaperone at the endoplasmic reticulum (ER) plays important roles in cellular regulation. Here we found a new function of Sig-1R, in that it translocates from the ER to the nuclear envelope (NE) to recruit chromatin-remodeling molecules and regulate the gene transcription thereof. Sig-1Rs mainly reside at the ER-mitochondrion interface. However, on stimulation by agonists such as cocaine, Sig-1Rs translocate from ER to the NE, where Sig-1Rs bind NE protein emerin and recruit chromatin-remodeling molecules, including lamin A/C, barrier-to-autointegration factor (BAF), and histone deacetylase (HDAC), to form a complex with the gene repressor specific protein 3 (Sp3). Knockdown of Sig-1Rs attenuates the complex formation. Cocaine was found to suppress the gene expression of monoamine oxidase B (MAOB) in the brain of wild-type but not Sig-1R knockout mouse. A single dose of cocaine (20 mg/kg) in rats suppresses the level of MAOB at nuclear accumbens without affecting the level of dopamine transporter. Daily injections of cocaine in rats caused behavioral sensitization. Withdrawal from cocaine in cocaine-sensitized rats induced an apparent time-dependent rebound of the MAOB protein level to about 200% over control on day 14 after withdrawal. Treatment of cocaine-withdrawn rats with the MAOB inhibitor deprenyl completely alleviated the behavioral sensitization to cocaine. Our results demonstrate a role of Sig-1R in transcriptional regulation and suggest cocaine may work through this newly discovered genomic action to achieve its addictive action. Results also suggest the MAOB inhibitor deprenyl as a therapeutic agent to block certain actions of cocaine during withdrawal.

  14. Sigma-1 receptor mediates cocaine-induced transcriptional regulation by recruiting chromatin-remodeling factors at the nuclear envelope

    PubMed Central

    Tsai, Shang-Yi A.; Chuang, Jian-Ying; Tsai, Meng-Shan; Wang, Xiao-fei; Hung, Jan-Jong; Chang, Wen-Chang; Bonci, Antonello; Su, Tsung-Ping

    2015-01-01

    The sigma-1 receptor (Sig-1R) chaperone at the endoplasmic reticulum (ER) plays important roles in cellular regulation. Here we found a new function of Sig-1R, in that it translocates from the ER to the nuclear envelope (NE) to recruit chromatin-remodeling molecules and regulate the gene transcription thereof. Sig-1Rs mainly reside at the ER–mitochondrion interface. However, on stimulation by agonists such as cocaine, Sig-1Rs translocate from ER to the NE, where Sig-1Rs bind NE protein emerin and recruit chromatin-remodeling molecules, including lamin A/C, barrier-to-autointegration factor (BAF), and histone deacetylase (HDAC), to form a complex with the gene repressor specific protein 3 (Sp3). Knockdown of Sig-1Rs attenuates the complex formation. Cocaine was found to suppress the gene expression of monoamine oxidase B (MAOB) in the brain of wild-type but not Sig-1R knockout mouse. A single dose of cocaine (20 mg/kg) in rats suppresses the level of MAOB at nuclear accumbens without affecting the level of dopamine transporter. Daily injections of cocaine in rats caused behavioral sensitization. Withdrawal from cocaine in cocaine-sensitized rats induced an apparent time-dependent rebound of the MAOB protein level to about 200% over control on day 14 after withdrawal. Treatment of cocaine-withdrawn rats with the MAOB inhibitor deprenyl completely alleviated the behavioral sensitization to cocaine. Our results demonstrate a role of Sig-1R in transcriptional regulation and suggest cocaine may work through this newly discovered genomic action to achieve its addictive action. Results also suggest the MAOB inhibitor deprenyl as a therapeutic agent to block certain actions of cocaine during withdrawal. PMID:26554014

  15. Advanced Paramagnetic Resonance Spectroscopies of Iron-Sulfur Proteins: Electron Nuclear Double Resonance (ENDOR) and Electron Spin Echo Envelope Modulation (ESEEM)

    PubMed Central

    Cutsail, George E.; Telser, Joshua; Hoffman, Brian M.

    2015-01-01

    The advanced electron paramagnetic resonance (EPR) techniques, electron nuclear double resonance (ENDOR) and electron spin echo envelope modulation (ESEEM) spectroscopies, provide unique insights into the structure, coordination chemistry, and biochemical mechanism of Nature’s widely distributed iron-sulfur cluster (FeS) proteins. This review describes the ENDOR and ESEEM techniques and then provides a series of case studies on their application to a wide variety of FeS proteins including ferredoxins, nitrogenase, and radical SAM enzymes. PMID:25686535

  16. Novel localization of formin mDia2: importin β-mediated delivery to and retention at the cytoplasmic side of the nuclear envelope

    PubMed Central

    Shao, Xiaowei; Kawauchi, Keiko; Shivashankar, G. V.; Bershadsky, Alexander D.

    2015-01-01

    ABSTRACT The formin family proteins are important regulators of actin polymerization that are involved in many cellular processes. However, little is known about their specific cellular localizations. Here, we show that Diaphanous-related formin-3 (mDia2) localizes to the cytoplasmic side of the nuclear envelope. This localization of mDia2 to the nuclear rim required the presence of a nuclear localization signal (NLS) sequence at the mDia2 N-terminal. Consistent with this result, super-resolution images demonstrated that at the nuclear rim, mDia2 co-localized with the nuclear pore complexes and a nuclear transport receptor, importin β. Furthermore, an interaction between mDia2 and importin β was detected by immunoprecipitation, and silencing of importin β was shown to attenuate accumulation of mDia2 to the nuclear rim. We have shown previously that Ca2+ entry leads to the assembly of perinuclear actin rim in an inverted formin 2 (INF2) dependent manner. mDia2, however, was not involved in this process since abolishing its localization at the nuclear rim by silencing of importin β had no effect on actin assembly at the nuclear rim triggered by Ca2+ stimulation. PMID:26519515

  17. SEPT12/SPAG4/LAMINB1 Complexes Are Required for Maintaining the Integrity of the Nuclear Envelope in Postmeiotic Male Germ Cells

    PubMed Central

    Yeh, Chung-Hsin; Kuo, Pao-Lin; Wang, Ya-Yun; Wu, Ying-Yu; Chen, Mei-Feng; Lin, Ding-Yen; Lai, Tsung-Hsuan; Chiang, Han-Sun; Lin, Ying-Hung

    2015-01-01

    Male infertility affects approximately 50% of all infertile couples. The male-related causes of intracytoplasmic sperm injection failure include the absence of sperm, immotile or immature sperm, and sperm with structural defects such as those caused by premature chromosomal condensation and DNA damage. Our previous studies based on a knockout mice model indicated that SEPT12 proteins are critical for the terminal morphological formation of sperm. SEPT12 mutations in men result in teratozospermia and oligozospermia. In addition, the spermatozoa exhibit morphological defects of the head and tail, premature chromosomal condensation, and nuclear damage. However, the molecular functions of SEPT12 during spermatogenesis remain unclear. To determine the molecular functions of SEPT12, we applied a yeast 2-hybrid system to identify SEPT12 interactors. Seven proteins that interact with SEPT12 were identified: SEPT family proteins (SEPT4 and SEPT6), nuclear or nuclear membrane proteins (protamine 2, sperm-associated antigen 4, and NDC1 transmembrane nucleoproine), and sperm-related structural proteins (pericentriolar material 1 and obscurin-like 1). Sperm-associated antigen 4 (SPAG4; also known as SUN4) belongs to the SUN family of proteins and acts as a linker protein between nucleoskeleton and cytoskeleton proteins and localizes in the nuclear membrane. We determined that SEPT12 interacts with SPAG4 in a male germ cell line through coimmunoprecipitation. During human spermiogenesis, SEPT12 is colocalized with SPAG4 near the nuclear periphery in round spermatids and in the centrosome region in elongating spermatids. Furthermore, we observed that SEPT12/SPAG4/LAMINB1 formed complexes and were coexpressed in the nuclear periphery of round spermatids. In addition, mutated SEPT12, which was screened from an infertile man, affected the integration of these nuclear envelope complexes through coimmunoprecipitation. This was the first study that suggested that SEPT proteins link to

  18. The nuclear pore complex protein ALADIN is anchored via NDC1 but not via POM121 and GP210 in the nuclear envelope

    SciTech Connect

    Kind, Barbara; Koehler, Katrin; Lorenz, Mike; Huebner, Angela

    2009-12-11

    The nuclear pore complex (NPC) consists of {approx}30 different proteins and provides the only sites for macromolecular transport between cytoplasm and nucleus. ALADIN was discovered as a new member of the NPC. Mutations in ALADIN are known to cause triple A syndrome, a rare autosomal recessive disorder characterized by adrenal insufficiency, alacrima, and achalasia. The function and exact location of the nucleoporin ALADIN within the NPC multiprotein complex is still unclear. Using a siRNA-based approach we downregulated the three known membrane integrated nucleoporins NDC1, GP210, and POM121 in stably expressing GFP-ALADIN HeLa cells. We identified NDC1 but not GP210 and POM121 as the main anchor of ALADIN within the NPC. Solely the depletion of NDC1 caused mislocalization of ALADIN. Vice versa, the depletion of ALADIN led also to disappearance of NDC1 at the NPC. However, the downregulation of two further membrane-integral nucleoporins GP210 and POM121 had no effect on ALADIN localization. Furthermore, we could show a direct association of NDC1 and ALADIN in NPCs by fluorescence resonance energy transfer (FRET) measurements. Based on our findings we conclude that ALADIN is anchored in the nuclear envelope via NDC1 and that this interaction gets lost, if ALADIN is mutated. The loss of integration of ALADIN in the NPC is a main pathogenetic aspect for the development of the triple A syndrome and suggests that the interaction between ALADIN and NDC1 may be involved in the pathogenesis of the disease.

  19. Task breakdown

    NASA Technical Reports Server (NTRS)

    Pavlich, Jane

    1990-01-01

    The topics concerning the Center for Space Construction (CSC) space construction breakdown structure are presented in viewgraph form. It is concluded that four components describe a task -- effecting, information gathering, analysis, and regulation; uncertainties effect the relative amount of information gathering and analysis that occurs; and that task timing requirements drive the 'location in time' of cognition.

  20. VAP-B binds to Rab3GAP1 at the ER: its implication in nuclear envelope formation through the ER-Golgi intermediate compartment.

    PubMed

    Hantan, Degejirihu; Yamamoto, Yasunori; Sakisaka, Toshiaki

    2014-01-01

    The vesicle-associated membrane protein-associated protein B (VAP-B) is a tail-anchored protein in the endoplasmic reticulum (ER). VAP-B functions as an adaptor protein to recruit target proteins to the ER and execute various cellular functions, lipid transport, membrane traffic, ER stress etc. Recently, VAP-B has been shown to regulate the nuclear envelope protein transport through the ER-Golgi intermediate compartment (ERGIC). We showed here that VAP-B directly binds to Rab3 GTPase activating protein 1 (Rab3GAP1), the catalytic subunit of Rab3GAP, through the two phenylalanines (FF) in an acidic tract (FFAT)-like motif of Rab3GAP1. Rab3GAP consists of two subunits, the catalytic subunit Rab3GAP1 and the non-catalytic subunit Rab3GAP2. VAP-B binds to Rab3GAP1 even in the Rab3GAP1/2 heterodimer complex. A single amino acid substitution of the FFAT-like motif reduces the binding activity of Rab3GAP1 to VAP-B. On the other hand, the FFAT-like motif mutation increases the binding activity of Rab3GAP1 to ERGIC-53, the ERGIC marker protein. Overexpression of Rab3GAP1 affects nuclear envelope formation more potently than that of Rab3GAP1 FFAT-like motif mutant. These results suggest that the binding of VAP-B to Rab3GAP1 is implicated in the regulation of nuclear envelope formation through ERGIC.

  1. Nuclear envelope-localized EGF family protein amphiregulin activates breast cancer cell migration in an EGF-like domain independent manner

    SciTech Connect

    Tanaka, Hisae; Nishioka, Yu; Yokoyama, Yuhki; Higashiyama, Shigeki; Matsuura, Nariaki; Matsuura, Shuji; Hieda, Miki

    2012-04-20

    Highlights: Black-Right-Pointing-Pointer Nuclear envelope-localized proAREG activates cancer cell migration via its cytoplasmic domain. Black-Right-Pointing-Pointer The induction of cell migration does not require the EGF-like domain or EGR function. Black-Right-Pointing-Pointer Nuclear envelope-localized proAREG suppresses breast cancer cell growth without EGFR function. Black-Right-Pointing-Pointer This study revealed a novel function mediated by the intracellular domain of proAREG. -- Abstract: Amphiregulin (AREG), an EGF family protein, is synthesized as a type I transmembrane precursor (proAREG) and expressed on the cell surface with an extracellular EGF-like domain and an intracellular short cytoplasmic tail. The ectodomain shedding yields a soluble EGF receptor ligand (soluble AREG) which binds to EGF receptor (EGFR) and concomitantly induces migration of unshed proAREG from the plasma membrane to the nuclear envelope (NE). AREG is known to play a potential role in breast cancer and has been intensively investigated as an EGF receptor ligand, while the function of the NE-localized proAREG remains unknown. In this study we used a truncated mutant that mimics NE-localized proAREG without shedding stimuli to discriminate between the functions of NE-localized and plasma membrane-localized proAREG and demonstrate that NE-localized proAREG activates breast cancer cell migration, but suppresses cell growth. Moreover, the present study shows that induction of cell migration by NE-localized proAREG does not require the extracellular growth factor domain or EGF receptor function. Collectively these data demonstrate a novel function mediated by the intracellular domain of proAREG and suggest a significant role for NE-localized proAREG in driving human breast cancer progression.

  2. Proteins from rat liver cytosol which stimulate mRNA transport. Purification and interactions with the nuclear envelope mRNA translocation system.

    PubMed

    Schröder, H C; Rottmann, M; Bachmann, M; Müller, W E; McDonald, A R; Agutter, P S

    1986-08-15

    Two polysome-associated proteins with particular affinities for poly(A) have been purified from rat liver. These proteins stimulate the efflux of mRNA from isolated nuclei in conditions under which such efflux closely stimulates mRNA transport in vivo, and they are therefore considered as mRNA-transport-stimulatory proteins. Their interaction with the mRNA-translocation system in isolated nuclear envelopes has been studied. The results are generally consistent with the most recently proposed kinetic model of mRNA translocation. One protein, P58, has not been described previously. It inhibits the protein kinase that down-regulates the NTPase, it enhances the NTPase activity in both the presence and the absence of poly(A) and it seems to increase poly(A) binding in unphosphorylated, but not in phosphorylated, envelopes. The other protein, P31, which probably corresponds to the 35,000-Mr factor described by Webb and his colleagues, enhances the binding of poly(A) to the mRNA-binding site in the envelope, thus stimulating the phosphoprotein phosphatase and, in consequence, the NTPase. The possible physiological significance of these two proteins is discussed.

  3. Advanced paramagnetic resonance spectroscopies of iron-sulfur proteins: Electron nuclear double resonance (ENDOR) and electron spin echo envelope modulation (ESEEM).

    PubMed

    Cutsail, George E; Telser, Joshua; Hoffman, Brian M

    2015-06-01

    The advanced electron paramagnetic resonance (EPR) techniques, electron nuclear double resonance (ENDOR) and electron spin echo envelope modulation (ESEEM) spectroscopies, provide unique insights into the structure, coordination chemistry, and biochemical mechanism of nature's widely distributed iron-sulfur cluster (FeS) proteins. This review describes the ENDOR and ESEEM techniques and then provides a series of case studies on their application to a wide variety of FeS proteins including ferredoxins, nitrogenase, and radical SAM enzymes. This article is part of a Special Issue entitled: Fe/S proteins: Analysis, structure, function, biogenesis and diseases.

  4. Immunological evidence for the localization of a 110 kDa poly(A) binding protein from rat liver in nuclear envelopes and its phosphorylation by protein kinase C.

    PubMed

    Schäfer, P; Aitken, S J; Bachmann, M; Agutter, P S; Müller, W E; Prochnow, D

    1993-11-01

    We have purified a 110 kDa poly(A) binding protein (P110) from rat liver which is thought to be involved in mRNA translocation through the nuclear pores and have demonstrated its localisation in the nuclear envelope using polyclonal antibodies and confocal laser scanning microscopy. Although P110 was prepared from highly purified nuclear envelopes, the polyclonal antibodies raised against them bind to nucleo- and cytoplasmic structures to a minor extent, but not to nucleolar structures. P110 decays spontaneously into several fragments which are also recognized by the polyclonal antibodies. The 110 kDa polypeptide and its fragments were phosphorylated by a nuclear envelope kinase and this phosphorylation was inhibited by a monoclonal antibody against protein kinase C and by a specific protein kinase C inhibitor obtained from bovine brain. Scatchard analysis was used to determine the influence of protein kinase C activators and inhibitors on nuclear envelope protein phosphorylation and RNA binding. The data indicate a close association between the RNA translocation machinery (the 110 kDa protein) and protein kinase C within the nuclear envelope. We suggest that the fragmentation of P110 is triggered before or during mRNA export and is not due to nonspecific proteolysis.

  5. A-type and B-type lamins initiate layer assembly at distinct areas of the nuclear envelope in living cells

    SciTech Connect

    Furukawa, Kazuhiro; Ishida, Kazuya; Tsunoyama, Taka-aki; Toda, Suguru; Osoda, Shinichi; Horigome, Tsuneyoshi; Fisher, Paul A.; Sugiyama, Shin

    2009-04-15

    To investigate nuclear lamina re-assembly in vivo, Drosophila A-type and B-type lamins were artificially expressed in Drosophila lamin Dm{sub 0}null mutant brain cells. Both exogenous lamin C (A-type) and Dm{sub 0} (B-type) formed sub-layers at the nuclear periphery, and efficiently reverted the abnormal clustering of the NPC. Lamin C initially appeared where NPCs were clustered, and subsequently extended along the nuclear periphery accompanied by the recovery of the regular distribution of NPCs. In contrast, lamin Dm{sub 0} did not show association with the clustered NPCs during lamina formation and NPC spacing recovered only after completion of a closed lamin Dm{sub 0} layer. Further, when lamin Dm{sub 0} and C were both expressed, they did not co-polymerize, initiating layer formation in separate regions. Thus, A and B-type lamins reveal differing properties during lamina assembly, with A-type having the primary role in organizing NPC distribution. This previously unknown complexity in the assembly of the nuclear lamina could be the basis for intricate nuclear envelope functions.

  6. Electrical Breakdown in Solids

    NASA Astrophysics Data System (ADS)

    Hjalmarson, Harold; Zutavern, Fred; Kambour, Kenneth; Moore, Chris; Mar, Alan

    During electron breakdown of a solid subjected to a large electric field, impact ionization causes growth of an electron-hole plasma. This growth process is opposed by Auger recombination of the electron-hole pairs. In our work, such breakdown is investigated by obtaining steady-state solutions to the Boltzmann equation. In these calculations, the carriers are heated by the electric field and cooled by phonon emission. Our results imply that breakdown may lead to high carrier-density current filaments. Conductive filaments have been observed in optically-triggered, high-power photoconductive semiconductor switch (PCSS) devices being developed at Sandia Labs. The relationship between the steady-state computed solutions to the observed filaments will be discussed in the presentation. This work was supported by the Laboratory Directed Research and Development program at Sandia National Laboratories. Sandia National Laboratories is a multi-program laboratory managed and operated by Sandia Corporation, a wholly owned subsidiary of Lockheed Martin Corporation, for the U.S. Department of Energy's National Nuclear Security Administration under Contract DE-AC04-94AL85000.

  7. The Use of Two-Photon FRET-FLIM to Study Protein Interactions During Nuclear Envelope Fusion In Vivo and In Vitro.

    PubMed

    Byrne, Richard D; Larijani, Banafshé; Poccia, Dominic L

    2016-01-01

    FRET-FLIM techniques have wide application in the study of protein and protein-lipid interactions in cells. We have pioneered an imaging platform for accurate detection of functional states of proteins and their interactions in fixed cells. This platform, two-site-amplified Förster resonance energy transfer (a-FRET), allows greater signal generation while retaining minimal noise thus enabling application of fluorescence lifetime imaging microscopy (FLIM) to be routinely deployed in different types of cells and tissue. We have used the method described here, time-resolved FRET monitored by two-photon FLIM, to demonstrate the direct interaction of Phospholipase Cγ (PLCγ) by Src Family Kinase 1 (SFK1) during nuclear envelope formation and during male and female pronuclear membrane fusion in fertilized sea urchin eggs. We describe here a generic method that can be applied to monitor any proteins of interest.

  8. The Use of Two-Photon FRET-FLIM to Study Protein Interactions During Nuclear Envelope Fusion In Vivo and In Vitro.

    PubMed

    Byrne, Richard D; Larijani, Banafshé; Poccia, Dominic L

    2016-01-01

    FRET-FLIM techniques have wide application in the study of protein and protein-lipid interactions in cells. We have pioneered an imaging platform for accurate detection of functional states of proteins and their interactions in fixed cells. This platform, two-site-amplified Förster resonance energy transfer (a-FRET), allows greater signal generation while retaining minimal noise thus enabling application of fluorescence lifetime imaging microscopy (FLIM) to be routinely deployed in different types of cells and tissue. We have used the method described here, time-resolved FRET monitored by two-photon FLIM, to demonstrate the direct interaction of Phospholipase Cγ (PLCγ) by Src Family Kinase 1 (SFK1) during nuclear envelope formation and during male and female pronuclear membrane fusion in fertilized sea urchin eggs. We describe here a generic method that can be applied to monitor any proteins of interest. PMID:27147038

  9. Lamin B1 Polymorphism Influences Morphology of the Nuclear Envelope, Cell Cycle Progression, and Risk of Neural Tube Defects in Mice

    PubMed Central

    De Castro, Sandra C. P.; Malhas, Ashraf; Leung, Kit-Yi; Gustavsson, Peter; Vaux, David J.; Copp, Andrew J.; Greene, Nicholas D. E.

    2012-01-01

    integrity of the nuclear envelope and ensuring normal cell cycle progression. PMID:23166514

  10. Evaluation of nitrogen nuclear hyperfine and quadrupole coupling parameters for the proximal imidazole in myoglobin-azide, -cyanide, and -mercaptoethanol complexes by electron spin echo envelope modulation spectroscopy.

    PubMed

    Magliozzo, R S; Peisach, J

    1993-08-24

    Electron spin echo envelope modulation (ESEEM) spectroscopy and computer simulation of spectra has been used to evaluate the nitrogen nuclear hyperfine and quadrupole coupling parameters for the proximal imidazole nitrogen directly coordinated to iron in three low-spin heme complexes, myoglobin-azide, -cyanide, and -mercaptoethanol (MbN3, MbCN, and MbRS). The variability in the weak electron-nuclear coupling parameters reveals the electronic flexibility within the heme group that depends on properties of the exogenous ligands. For example, the isotropic component of the nitrogen nuclear hyperfine coupling ranges from 4.4 MHz for MbN3 to 2.2 MHz for both MbCN and MbRS. The weaker coupling in MbCN and MbRS is taken as evidence for delocalization of unpaired electron spin from iron into the exogenous anionic ligands. The value of e2Qq, the nuclear quadrupole coupling constant for the axial imidazole nitrogen in MbCN and MbRS, was 2.5 MHz but was significantly larger, 3.2 MHz, in MbN3. This large value is considered evidence for a weakened sigma bond between the proximal imidazole and ferric iron in this form, and for a feature contributing to the origin of the high spin-low spin equilibrium exhibited by MbN3 [Beetlestone, J., & George, P. (1964) Biochemistry 5, 707-714]. The ESEEM results have allowed a correlation to be made between the orientation of the g tensor axes, the orientation of the p-pi orbital of the proximal imidazole nitrogen, and sigma- and pi-bonding features of the axial ligands. Furthermore, the proximal imidazole is suggested to act as a pi-acceptor in low-spin heme complexes in order to support strong sigma electron donation from the lone pair orbital to iron. An evaluation of the nitrogen nuclear hyperfine coupling parameters for the porphyrin pyrrole sites in MbRS reveals a large inequivalence in isotropic components consistent with an orientation of rhombic axes (and g tensor axes) that eclipses the Fe-Npyrrole vector directions. PMID:8395204

  11. DC Breakdown Experiments

    SciTech Connect

    Calatroni, S.; Descoeudres, A.; Levinsen, Y.; Taborelli, M.; Wuensch, W.

    2009-01-22

    In the context of the CLIC (Compact Linear Collider) project investigations of DC breakdown in ultra high vacuum are carried out in parallel with high power RF tests. From the point of view of saturation breakdown field the best material tested so far is stainless steel, followed by titanium. Copper shows a four times weaker breakdown field than stainless steel. The results indicate clearly that the breakdown events are initiated by field emission current and that the breakdown field is limited by the cathode. In analogy to RF, the breakdown probability has been measured in DC and the data show similar behaviour as a function of electric field.

  12. Nuclear envelope precursor vesicle targeting to chromatin is stimulated by protein phosphatase 1 in Xenopus egg extracts

    SciTech Connect

    Ito, Hiromi; Koyama, Yuhei; Takano, Makoto; Ishii, Kohei; Maeno, Mitsugu; Furukawa, Kazuhiro; Horigome, Tsuneyoshi . E-mail: thori@chem.sc.niigata-u.ac.jp

    2007-05-15

    The mechanism underlying targeting of the nuclear membrane to chromatin at the end of mitosis was studied using an in vitro cell-free system comprising Xenopus egg membrane and cytosol fractions, and sperm chromatin. The mitotic phase membrane, which was separated from a mitotic phase extract of Xenopus eggs and could not bind to chromatin, became able to bind to chromatin on pretreatment with a synthetic phase cytosol fraction of Xenopus eggs. When the cytosol fraction was depleted of protein phosphatase 1 (PP1) with anti-Xenopus PP1{gamma}1 antibodies, this ability was lost. The addition of recombinant xPP1{gamma}1 to the PP1-depleted cytosol fraction restored the ability. These and other results suggested that dephosphorylation of mitotic phosphorylation sites on membranes by PP1 in the synthetic phase cytosol fraction promoted targeting of the membranes to chromatin. On the other hand, a fragment containing the chromatin-binding domain of lamin B receptor (LBR) but not emerin inhibited targeting of membrane vesicles. It was also shown that PP1 dephosphorylates a phosphate group(s) responsible for regulation of the binding of LBR to chromatin. A possible mechanism involving PP1 and LBR for the regulation of nuclear membrane targeting to chromatin was discussed.

  13. Nuclear envelope proteins Nesprin2 and LaminA regulate proliferation and apoptosis of vascular endothelial cells in response to shear stress.

    PubMed

    Han, Yue; Wang, Lu; Yao, Qing-Ping; Zhang, Ping; Liu, Bo; Wang, Guo-Liang; Shen, Bao-Rong; Cheng, Binbin; Wang, Yingxiao; Jiang, Zong-Lai; Qi, Ying-Xin

    2015-05-01

    The dysfunction of vascular endothelial cells (ECs) influenced by flow shear stress is crucial for vascular remodeling. However, the roles of nuclear envelope (NE) proteins in shear stress-induced EC dysfunction are still unknown. Our results indicated that, compared with normal shear stress (NSS), low shear stress (LowSS) suppressed the expression of two types of NE proteins, Nesprin2 and LaminA, and increased the proliferation and apoptosis of ECs. Targeted small interfering RNA (siRNA) and gene overexpression plasmid transfection revealed that Nesprin2 and LaminA participate in the regulation of EC proliferation and apoptosis. A protein/DNA array was further used to detect the activation of transcription factors in ECs following transfection with target siRNAs and overexpression plasmids. The regulation of AP-2 and TFIID mediated by Nesprin2 and the activation of Stat-1, Stat-3, Stat-5 and Stat-6 by LaminA were verified under shear stress. Furthermore, using Ingenuity Pathway Analysis software and real-time RT-PCR, the effects of Nesprin2 or LaminA on the downstream target genes of AP-2, TFIID, and Stat-1, Stat-3, Stat-5 and Stat-6, respectively, were investigated under LowSS. Our study has revealed that NE proteins are novel mechano-sensitive molecules in ECs. LowSS suppresses the expression of Nesprin2 and LaminA, which may subsequently modulate the activation of important transcription factors and eventually lead to EC dysfunction.

  14. Effects of globularifolin on cell survival, nuclear factor-κB activity, neopterin production, tryptophan breakdown and free radicals in vitro.

    PubMed

    Sipahi, Hande; Becker, Kathrin; Gostner, Johanna M; Charehsaz, Mohammad; Kirmizibekmez, Hasan; Schennach, Harald; Aydin, Ahmet; Fuchs, Dietmar

    2014-01-01

    The potential effects of globularifolin, an acylated iridoid glucoside, on cell survival, inflammation markers and free radicals scavenging were investigated. Viability assay on human myelomomonocytic cell line THP-1 and human peripheral blood mononuclear cells (PBMC) using the Cell-Titer Blue assay proved that globularifolin had no toxic effect at the tested concentrations. Conversely, it is proportional to the dose globularifolin increased growth of THP-1 cells (p <0.01). On human PBMC, globularifolin at 6.25 and 12.5 μM concentrations showed a stimulatory effect, while at 12.5-200 μM it suppressed response of PBMC to stimulation with phytohemagglutinin (PHA). Globularifolin (50-200 μM) enhanced neopterin formation dose-dependently, whereas tryptophan breakdown was not influenced. At 50-200 μM in unstimulated PBMC in THP-1 cells, globularifolin induced a significant expression of nuclear factor-κB (NF-κB) as was quantified by Quanti-Blue assay. By contrast, in lipopolysaccharide (LPS)-stimulated cells, the higher concentrations of globularifolin suppressed NF-κB expression dose-dependently and a significant decrease was observed at 200 μM concentration. A positive correlation was found between increased neopterin and NF-κB activity (p <0.01). Similarly, a positive correlation was observed between neopterin levels in mitogen-induced cells and NF-κB activity in LPS-stimulated cells after treatment with globularifolin (p=0.001). The free radical scavenging capacity of globularifolin evaluated by Oxygen Radical Absorbance Capacity (ORAC) assay showed relative ORAC values of 0.36±0.05 μmol Trolox equivalent/μmol. All together, results show that natural antioxidant globularifolin might represent a potential immunomodulatory as well as proliferative agent, which deserves further in vitro and in vivo studies. PMID:24185011

  15. Electrical breakdown of nanowires.

    PubMed

    Zhao, Jiong; Sun, Hongyu; Dai, Sheng; Wang, Yan; Zhu, Jing

    2011-11-01

    Instantaneous electrical breakdown measurements of GaN and Ag nanowires are performed by an in situ transmission electron microscopy method. Our results directly reveal the mechanism that typical thermally heated semiconductor nanowires break at the midpoint, while metallic nanowires breakdown near the two ends due to the stress induced by electromigration. The different breakdown mechanisms for the nanowires are caused by the different thermal and electrical properties of the materials.

  16. Spectrin repeat containing nuclear envelope 1 and forkhead box protein E1 are promising markers for the detection of colorectal cancer in blood.

    PubMed

    Melotte, Veerle; Yi, Joo Mi; Lentjes, Marjolein H F M; Smits, Kim M; Van Neste, Leander; Niessen, Hanneke E C; Wouters, Kim A D; Louwagie, Joost; Schuebel, Kornel E; Herman, James G; Baylin, Stephen B; van Criekinge, Wim; Meijer, Gerrit A; Ahuja, Nita; van Engeland, Manon

    2015-02-01

    Identifying biomarkers in body fluids may improve the noninvasive detection of colorectal cancer. Previously, we identified N-Myc downstream-regulated gene 4 (NDRG4) and GATA binding protein 5 (GATA5) methylation as promising biomarkers for colorectal cancer in stool DNA. Here, we examined the utility of NDRG4, GATA5, and two additional markers [Forkhead box protein E1 (FOXE1) and spectrin repeat containing nuclear envelope 1 (SYNE1)] promoter methylation as biomarkers in plasma DNA. Quantitative methylation-specific PCR was performed on plasma DNA from 220 patients with colorectal cancer and 684 noncancer controls, divided in a training set and a test set. Receiver operating characteristic analysis was performed to measure the area under the curve of GATA5, NDRG4, SYNE1, and FOXE1 methylation. Functional assays were performed in SYNE1 and FOXE1 stably transfected cell lines. The sensitivity of NDRG4, GATA5, FOXE1, and SYNE1 methylation in all stages of colorectal cancer (154 cases, 444 controls) was 27% [95% confidence interval (CI), 20%-34%), 18% (95% CI, 12%-24%), 46% (95% CI, 38%-54%), and 47% (95% CI, 39%-55%), with a specificity of 95% (95% CI, 93%-97%), 99% (95% CI, 98%-100%), 93% (95% CI, 91%-95%), and 96% (95% CI, 94%-98%), respectively. Combining SYNE1 and FOXE1, increased the sensitivity to 56% (95% CI, 48%-64%), while the specificity decreased to 90% (95% CI, 87%-93%) in the training set and to 58% sensitivity (95% CI, 46%-70%) and 91% specificity (95% CI, 80%-100%) in a test set (66 cases, 240 controls). SYNE1 overexpression showed no major differences in cell proliferation, migration, and invasion compared with controls. Overexpression of FOXE1 significantly decreased the number of colonies in SW480 and HCT116 cell lines. Overall, our data suggest that SYNE1 and FOXE1 are promising markers for colorectal cancer detection. PMID:25538088

  17. Vortex breakdown simulation

    NASA Technical Reports Server (NTRS)

    Nakamura, Y.; Leonard, A.; Spalart, P. R.

    1985-01-01

    A vortex breakdown was simulated by the vortex filament method, and detailed figures are presented based on the results. Deformations of the vortex filaments showed clear and large swelling at a particular axial station which implied the presence of a recirculation bubble at that station. The tendency for two breakdowns to occur experimentally was confirmed by the simulation, and the jet flow inside the bubble was well simulated. The particle paths spiralled with expansion, and the streamlines took spiral forms at the breakdown with expansion.

  18. Dynamics of Sun5 Localization during Spermatogenesis in Wild Type and Dpy19l2 Knock-Out Mice Indicates That Sun5 Is Not Involved in Acrosome Attachment to the Nuclear Envelope

    PubMed Central

    Yassine, Sandra; Escoffier, Jessica; Nahed, Roland Abi; Pierre, Virginie; Karaouzene, Thomas; Ray, Pierre F.; Arnoult, Christophe

    2015-01-01

    The acrosome is an organelle that is central to sperm physiology and a defective acrosome biogenesis leads to globozoospermia, a severe male infertility. The identification of the actors involved in acrosome biogenesis is therefore particularly important to decipher the molecular pathogeny of globozoospermia. We recently showed that a defect in the DPY19L2 gene is present in more than 70% of globozoospermic men and demonstrated that Dpy19l2, located in the inner nuclear membrane, is the first protein involved in the attachment of the acrosome to the nuclear envelope (NE). SUN proteins serve to link the nuclear envelope to the cytoskeleton and are therefore good candidates to participate in acrosome-nucleus attachment, potentially by interacting with DPY19L2. In order to characterize new actors of acrosomal attachment, we focused on Sun5 (also called Spag4l), which is highly expressed in male germ cells, and investigated its localization during spermatogenesis. Using immunohistochemistry and Western blot experiments in mice, we showed that Sun5 transits through different cellular compartments during meiosis. In pachytene spermatocytes, it is located in a membranous compartment different to the reticulum. In round spermatids, it progresses to the Golgi and the NE before to be located to the tail/head junction in epididymal sperm. Interestingly, we demonstrate that Sun5 is not, as initially reported, facing the acrosome but is in fact excluded from this zone. Moreover, we show that in Dpy19l2 KO spermatids, upon the detachment of the acrosome, Sun5 relocalizes to the totality of the NE suggesting that the acrosome attachment excludes Sun5 from the NE facing the acrosome. Finally, Western-blot experiments demonstrate that Sun5 is glycosylated. Overall, our work, associated with other publications, strongly suggests that the attachment of the acrosome to the nucleus does not likely depend on the formation of SUN complexes. PMID:25775128

  19. The Bacterial Cell Envelope

    PubMed Central

    Silhavy, Thomas J.; Kahne, Daniel; Walker, Suzanne

    2010-01-01

    The bacteria cell envelope is a complex multilayered structure that serves to protect these organisms from their unpredictable and often hostile environment. The cell envelopes of most bacteria fall into one of two major groups. Gram-negative bacteria are surrounded by a thin peptidoglycan cell wall, which itself is surrounded by an outer membrane containing lipopolysaccharide. Gram-positive bacteria lack an outer membrane but are surrounded by layers of peptidoglycan many times thicker than is found in the Gram-negatives. Threading through these layers of peptidoglycan are long anionic polymers, called teichoic acids. The composition and organization of these envelope layers and recent insights into the mechanisms of cell envelope assembly are discussed. PMID:20452953

  20. Mechanism of dissolution of envelopes of the extreme halophile Halobacterium cutirubrum.

    PubMed

    Onishi, H; Kushner, D J

    1966-02-01

    Onishi, H. (National Research Council, Ottawa, Ontario, Canada), and D. J. Kushner. Mechanism of dissolution of envelopes of the extreme halophile Halobacterium cutirubrum. J. Bacteriol. 91:646-652. 1966.-Envelopes of Halobacterium cutirubrum dissolved rapidly in media of low ionic strength. Heating partially inhibited breakdown, probably because of nonspecific protein coagulation rather than inactivation of a lytic enzyme(s). Dissolution of envelopes in water did not involve splitting of peptide bonds or protein-lipid bonds, or any extensive breakdown of carbohydrate polymers. Dissolution was increased by alcohols and urea, even at high salt concentrations, but was not affected by metabolic inhibitors. Thus, no evidence was found for a dilution-activated lytic enzyme that contributes to envelope breakdown. Cells of H. cutirubrum were stable in 2 m NaCl, but lysis occurred in 2 m KCl or NH(4)Cl. This lysis did not involve an extensive breakdown of the envelope. No evidence for different sites of Na(+), K(+), and NH(4) (+) action was obtained from the pattern of release of envelope constituents in different concentrations of these salts. Ultracentrifugation studies showed that adding salts to envelopes that had been dissolved in water led to a nonspecific reaggregation of envelope material. No difference was seen between the effects of KCl and NaCl, except at 3 to 4 m concentrations where KCl caused more aggregation. The preferential effect of Na(+) on intact cells is probably due to its ability specifically to prevent leakage rather than to an overall effect on envelope integrity.

  1. Beauty in the Breakdown

    ERIC Educational Resources Information Center

    Brisco, Nicole

    2008-01-01

    Most human beings look at erosion as the destruction of a surface, but artists can see that erosion often creates indefinable beauty. Where do you see beauty in the breakdown? In this article, the author presents an innovative lesson that would allow students to observe both human and physical nature. In this activity students will create a work…

  2. Measuring Breakdown Voltage.

    ERIC Educational Resources Information Center

    Auer, Herbert J.

    1978-01-01

    The article discusses an aspect of conductivity, one of the electrical properties subdivisions, and describes a tester that can be shop-built. Breakdown voltage of an insulation material is specifically examined. Test procedures, parts lists, diagrams, and test data form are included. (MF)

  3. SAFEGUARDS ENVELOPE: PREVIOUS WORK AND EXAMPLES

    SciTech Connect

    Richard Metcalf; Aaron Bevill; William Charlton; Robert Bean

    2008-07-01

    The future expansion of nuclear power will require not just electricity production but fuel cycle facilities such as fuel fabrication and reprocessing plants. As large reprocessing facilities are built in various states, they must be built and operated in a manner to minimize the risk of nuclear proliferation. Process monitoring has returned to the spotlight as an added measure that can increase confidence in the safeguards of special nuclear material (SNM). Process monitoring can be demonstrated to lengthen the allowable inventory period by reducing accountancy requirements, and to reduce the false positive indications. The next logical step is the creation of a Safeguards Envelope, a set of operational parameters and models to maximize anomaly detection and inventory period by process monitoring while minimizing operator impact and false positive rates. A brief example of a rudimentary Safeguards Envelope is presented, and shown to detect synthetic diversions overlaying a measured processing plant data set. This demonstration Safeguards Envelope is shown to increase the confidence that no SNM has been diverted with minimal operator impact, even though it is based on an information sparse environment. While the foundation on which a full Safeguards Envelope can be built has been presented in historical demonstrations of process monitoring, several requirements remain yet unfulfilled. Future work will require reprocessing plant transient models, inclusion of “non-traditional” operating data, and exploration of new methods of identifying subtle events in transient processes.

  4. Effects of Deletion and Overexpression of the Autographa californica Nuclear Polyhedrosis Virus FP25K Gene on Synthesis of Two Occlusion-Derived Virus Envelope Proteins and Their Transport into Virus-Induced Intranuclear Membranes

    PubMed Central

    Rosas-Acosta, Germán; Braunagel, Sharon C.; Summers, Max D.

    2001-01-01

    Partial deletions within Autographa californica open reading frame 61 (FP25K) alter the expression and accumulation profile of several viral proteins and the transport of occlusion-derived virus (ODV)-E66 to intranuclear membranes during infection (S. C. Braunagel et al., J. Virol. 73:8559–8570, 1999). Here we show the effects of a full deletion and overexpression of FP25K on the transport and expression of two ODV envelope proteins, ODV-E66 (E66) and ODV-E25 (E25). Deletion and overexpression of FP25K substantially altered the levels of expression of E66 during infection. Compared with cells infected with wild-type (wt) virus, the levels of E66 were reduced fivefold in cells infected with a viral mutant lacking FP25K (ΔFP25K) and were slightly increased in cells infected with a viral mutant overexpressing FP25K (FP25Kpolh). In contrast, no significant changes were observed in the levels of E25 among wt-, ΔFP25K-, and FP25Kpolh-infected cells. The changes observed in the levels of E66 among the different viral mutants were not accompanied by changes in either the time of synthesis, membrane association, protein turnover, or steady-state transcript abundance. Deletion of FP25K also substantially altered the transport and localization of E66 during infection. In cells infected with the ΔFP25K mutant virus, E66 accumulated in localized regions at the nuclear periphery and the outer nuclear membrane and did not traffic to intranuclear membranes. In contrast, in cells infected with the FP25Kpolh mutant virus E66 trafficked to intranuclear membranes. For comparison, E25 was normally transported to intranuclear membranes in both ΔFP25K- and FP25Kpolh-infected cells. Altogether these studies suggest that FP25K affects the synthesis of E66 at a posttranscriptional level, probably by altering the translation of E66; additionally, the block in transport of E66 at the nuclear envelope in ΔFP25K-infected cells suggests that the pathway of E66 trafficking to the inner

  5. Drug design from the cryptic inhibitor envelope

    PubMed Central

    Lee, Chul-Jin; Liang, Xiaofei; Wu, Qinglin; Najeeb, Javaria; Zhao, Jinshi; Gopalaswamy, Ramesh; Titecat, Marie; Sebbane, Florent; Lemaitre, Nadine; Toone, Eric J.; Zhou, Pei

    2016-01-01

    Conformational dynamics plays an important role in enzyme catalysis, allosteric regulation of protein functions and assembly of macromolecular complexes. Despite these well-established roles, such information has yet to be exploited for drug design. Here we show by nuclear magnetic resonance spectroscopy that inhibitors of LpxC—an essential enzyme of the lipid A biosynthetic pathway in Gram-negative bacteria and a validated novel antibiotic target—access alternative, minor population states in solution in addition to the ligand conformation observed in crystal structures. These conformations collectively delineate an inhibitor envelope that is invisible to crystallography, but is dynamically accessible by small molecules in solution. Drug design exploiting such a hidden inhibitor envelope has led to the development of potent antibiotics with inhibition constants in the single-digit picomolar range. The principle of the cryptic inhibitor envelope approach may be broadly applicable to other lead optimization campaigns to yield improved therapeutics. PMID:26912110

  6. FRACTIONAL CRYSTALLIZATION FEED ENVELOPE

    SciTech Connect

    HERTING DL

    2008-03-19

    Laboratory work was completed on a set of evaporation tests designed to establish a feed envelope for the fractional crystallization process. The feed envelope defines chemical concentration limits within which the process can be operated successfully. All 38 runs in the half-factorial design matrix were completed successfully, based on the qualitative definition of success. There is no feed composition likely to be derived from saltcake dissolution that would cause the fractional crystallization process to not meet acceptable performance requirements. However, some compositions clearly would provide more successful operation than other compositions.

  7. Safeguards Envelope Progress FY10

    SciTech Connect

    Richard Metcalf

    2010-10-01

    The Safeguards Envelope is a strategy to determine a set of specific operating parameters within which nuclear facilities may operate to maximize safeguards effectiveness without sacrificing safety or plant efficiency. This paper details the additions to the advanced operating techniques that will be applied to real plant process monitoring (PM) data from the Idaho Chemical Processing Plant (ICPP). Research this year focused on combining disparate pieces of data together to maximize operating time with minimal downtime due to safeguards. A Chi-Square and Croiser's cumulative sum were both included as part of the new analysis. Because of a major issue with the original data, the implementation of the two new tests did not add to the existing set of tests, though limited one-variable optimization made a small increase in detection probability. Additional analysis was performed to determine if prior analysis would have caused a major security or safety operating envelope issue. It was determined that a safety issue would have resulted from the prior research, but that the security may have been increased under certain conditions.

  8. Space Charge Modulated Electrical Breakdown

    PubMed Central

    Li, Shengtao; Zhu, Yuanwei; Min, Daomin; Chen, George

    2016-01-01

    Electrical breakdown is one of the most important physical phenomena in electrical and electronic engineering. Since the early 20th century, many theories and models of electrical breakdown have been proposed, but the origin of one key issue, that the explanation for dc breakdown strength being twice or higher than ac breakdown strength in insulating materials, remains unclear. Here, by employing a bipolar charge transport model, we investigate the space charge dynamics in both dc and ac breakdown processes. We demonstrate the differences in charge accumulations under both dc and ac stresses and estimate the breakdown strength, which is modulated by the electric field distortion induced by space charge. It is concluded that dc breakdown initializes in the bulk whereas ac breakdown initializes in the vicinity of the sample-electrode interface. Compared with dc breakdown, the lower breakdown strength under ac stress and the decreasing breakdown strength with an increase in applied frequency, are both attributed to the electric field distortion induced by space charges located in the vicinity of the electrodes. PMID:27599577

  9. Space Charge Modulated Electrical Breakdown

    NASA Astrophysics Data System (ADS)

    Li, Shengtao; Zhu, Yuanwei; Min, Daomin; Chen, George

    2016-09-01

    Electrical breakdown is one of the most important physical phenomena in electrical and electronic engineering. Since the early 20th century, many theories and models of electrical breakdown have been proposed, but the origin of one key issue, that the explanation for dc breakdown strength being twice or higher than ac breakdown strength in insulating materials, remains unclear. Here, by employing a bipolar charge transport model, we investigate the space charge dynamics in both dc and ac breakdown processes. We demonstrate the differences in charge accumulations under both dc and ac stresses and estimate the breakdown strength, which is modulated by the electric field distortion induced by space charge. It is concluded that dc breakdown initializes in the bulk whereas ac breakdown initializes in the vicinity of the sample-electrode interface. Compared with dc breakdown, the lower breakdown strength under ac stress and the decreasing breakdown strength with an increase in applied frequency, are both attributed to the electric field distortion induced by space charges located in the vicinity of the electrodes.

  10. Space Charge Modulated Electrical Breakdown.

    PubMed

    Li, Shengtao; Zhu, Yuanwei; Min, Daomin; Chen, George

    2016-01-01

    Electrical breakdown is one of the most important physical phenomena in electrical and electronic engineering. Since the early 20(th) century, many theories and models of electrical breakdown have been proposed, but the origin of one key issue, that the explanation for dc breakdown strength being twice or higher than ac breakdown strength in insulating materials, remains unclear. Here, by employing a bipolar charge transport model, we investigate the space charge dynamics in both dc and ac breakdown processes. We demonstrate the differences in charge accumulations under both dc and ac stresses and estimate the breakdown strength, which is modulated by the electric field distortion induced by space charge. It is concluded that dc breakdown initializes in the bulk whereas ac breakdown initializes in the vicinity of the sample-electrode interface. Compared with dc breakdown, the lower breakdown strength under ac stress and the decreasing breakdown strength with an increase in applied frequency, are both attributed to the electric field distortion induced by space charges located in the vicinity of the electrodes. PMID:27599577

  11. Proteolysis of Xenopus laevis egg envelope ZPA triggers envelope hardening.

    PubMed

    Lindsay, Leann L; Hedrick, Jerry L

    2004-11-12

    The egg envelope of most animal eggs is modified following fertilization, resulting in the prevention of polyspermy and hardening of the egg envelope. In frogs and mammals a prominent feature of envelope modification is N-terminal proteolysis of the envelope glycoprotein ZPA. We have purified the ZPA protease from Xenopus laevis eggs and characterized it as a zinc metalloprotease. Proteolysis of isolated egg envelopes by the isolated protease resulted in envelope hardening. The N-terminal peptide fragment of ZPA remained disulfide bond linked to the ZPA glycoprotein moiety following proteolysis. We propose a mechanism for egg envelope hardening involving ZPA proteolysis by an egg metalloprotease as a triggering event followed by induction of global conformational changes in egg envelope glycoproteins. PMID:15474476

  12. Jacketed lamp bulb envelope

    DOEpatents

    MacLennan, Donald A.; Turner, Brian P.; Gitsevich, Aleksandr; Bass, Gary K.; Dolan, James T.; Kipling, Kent; Kirkpatrick, Douglas A.; Leng, Yongzhang; Levin, Izrail; Roy, Robert J.; Shanks, Bruce; Smith, Malcolm; Trimble, William C.; Tsai, Peter

    2001-01-01

    A jacketed lamp bulb envelope includes a ceramic cup having an open end and a partially closed end, the partially closed end defining an aperture, a lamp bulb positioned inside the ceramic cup abutting the aperture, and a reflective ceramic material at least partially covering a portion of the bulb not abutting the aperture. The reflective ceramic material may substantially fill an interior volume of the ceramic cup not occupied by the bulb. The ceramic cup may include a structural feature for aiding in alignment of the jacketed lamp bulb envelope in a lamp. The ceramic cup may include an external flange about a periphery thereof. One example of a jacketed lamp bulb envelope includes a ceramic cup having an open end and a closed end, a ceramic washer covering the open end of the ceramic cup, the washer defining an aperture therethrough, a lamp bulb positioned inside the ceramic cup abutting the aperture, and a reflective ceramic material filling an interior volume of the ceramic cup not occupied by the bulb. A method of packing a jacketed lamp bulb envelope of the type comprising a ceramic cup with a lamp bulb disposed therein includes the steps of filling the ceramic cup with a flowable slurry of reflective material, and applying centrifugal force to the cup to pack the reflective material therein.

  13. Pushing the endogenous envelope

    PubMed Central

    Henzy, Jamie E.; Johnson, Welkin E.

    2013-01-01

    The majority of retroviral envelope glycoproteins characterized to date are typical of type I viral fusion proteins, having a receptor binding subunit associated with a fusion subunit. The fusion subunits of lentiviruses and alpha-, beta-, delta- and gammaretroviruses have a very conserved domain organization and conserved features of secondary structure, making them suitable for phylogenetic analyses. Such analyses, along with sequence comparisons, reveal evidence of numerous recombination events in which retroviruses have acquired envelope glycoproteins from heterologous sequences. Thus, the envelope gene (env) can have a history separate from that of the polymerase gene (pol), which is the most commonly used gene in phylogenetic analyses of retroviruses. Focusing on the fusion subunits of the genera listed above, we describe three distinct types of retroviral envelope glycoproteins, which we refer to as gamma-type, avian gamma-type and beta-type. By tracing these types within the ‘fossil record’ provided by endogenous retroviruses, we show that they have surprisingly distinct evolutionary histories and dynamics, with important implications for cross-species transmissions and the generation of novel lineages. These findings validate the utility of env sequences in contributing phylogenetic signal that enlarges our understanding of retrovirus evolution. PMID:23938755

  14. COMMON ENVELOPE: ENTHALPY CONSIDERATION

    SciTech Connect

    Ivanova, N.; Chaichenets, S.

    2011-04-20

    In this Letter, we discuss a modification to the criterion for the common envelope (CE) event to result in envelope dispersion. We emphasize that the current energy criterion for the CE phase is not sufficient for an instability of the CE, nor for an ejection. However, in some cases, stellar envelopes undergo stationary mass outflows, which are likely to occur during the slow spiral-in stage of the CE event. We propose the condition for such outflows, in a manner similar to the currently standard {alpha}{sub CE}{lambda}-prescription but with an addition of P/{rho} term in the energy balance equation, accounting therefore for the enthalpy of the envelope rather than merely the gas internal energy. This produces a significant correction, which might help to dispense with an unphysically high value of energy efficiency parameter during the CE phase, currently required in the binary population synthesis studies to make the production of low-mass X-ray binaries with a black hole companion to match the observations.

  15. The nuclear envelopathies and human diseases

    PubMed Central

    2009-01-01

    The nuclear envelope (NE) consists of two membrane layers that segregate the nuclear from the cytoplasmic contents. Recent progress in our understanding of nuclear-lamina associated diseases has revealed intriguing connections between the envelope components and nuclear processes. Here, we review the functions of the nuclear envelope in chromosome organization, gene expression, DNA repair and cell cycle progression, and correlate deficiencies in envelope function with human pathologies. PMID:19849840

  16. Exclusion of mRNPs and ribosomal particles from a thin zone beneath the nuclear envelope revealed upon inhibition of transport

    SciTech Connect

    Kylberg, Karin; Bjoerk, Petra; Fomproix, Nathalie; Ivarsson, Birgitta; Wieslander, Lars; Daneholt, Bertil

    2010-04-01

    We have studied the nucleocytoplasmic transport of a specific messenger RNP (mRNP) particle, named Balbiani ring (BR) granule, and ribosomal RNP (rRNP) particles in the salivary glands of the dipteran Chironomus tentans. The passage of the RNPs through the nuclear pore complex (NPC) was inhibited with the nucleoporin-binding wheat germ agglutinin, and the effects were examined by electron microscopy. BR mRNPs bound to the nuclear basket increased in number, while BR mRNPs translocating through the central channel decreased, suggesting that the initiation of translocation proper had been inhibited. The rRNPs accumulated heavily in nucleoplasm, while no or very few rRNPs were recorded within nuclear baskets. Thus, the transport of rRNPs had been blocked prior to the entry into the baskets. Remarkably, the rRNPs had been excluded both from baskets and the space in between the baskets. We propose that normally basket fibrils move freely and repel RNPs from the exclusion zone unless the particles have affinity for and bind to nucleoporins within the baskets.

  17. Exclusion of mRNPs and ribosomal particles from a thin zone beneath the nuclear envelope revealed upon inhibition of transport.

    PubMed

    Kylberg, Karin; Björk, Petra; Fomproix, Nathalie; Ivarsson, Birgitta; Wieslander, Lars; Daneholt, Bertil

    2010-04-01

    We have studied the nucleocytoplasmic transport of a specific messenger RNP (mRNP) particle, named Balbiani ring (BR) granule, and ribosomal RNP (rRNP) particles in the salivary glands of the dipteran Chironomus tentans. The passage of the RNPs through the nuclear pore complex (NPC) was inhibited with the nucleoporin-binding wheat germ agglutinin, and the effects were examined by electron microscopy. BR mRNPs bound to the nuclear basket increased in number, while BR mRNPs translocating through the central channel decreased, suggesting that the initiation of translocation proper had been inhibited. The rRNPs accumulated heavily in nucleoplasm, while no or very few rRNPs were recorded within nuclear baskets. Thus, the transport of rRNPs had been blocked prior to the entry into the baskets. Remarkably, the rRNPs had been excluded both from baskets and the space in between the baskets. We propose that normally basket fibrils move freely and repel RNPs from the exclusion zone unless the particles have affinity for and bind to nucleoporins within the baskets.

  18. Work breakdown structure guide

    SciTech Connect

    Not Available

    1987-02-06

    Utilization of the work breakdown structure (WBS) technique is an effective aid in managing Department of Energy (DOE) programs and projects. The technique provides a framework for project management by focusing on the products that are being developed or constructed to solve technical problems. It assists both DOE and contractors in fulfilling their management responsibilities. This document provides guidance for use of the WBS technique for product oriented work identification and definition. It is one in a series of policy and guidance documents supporting DOE's project manaagement system.

  19. Comparative genomics in Chlamydomonas and Plasmodium identifies an ancient nuclear envelope protein family essential for sexual reproduction in protists, fungi, plants, and vertebrates

    PubMed Central

    Ning, Jue; Otto, Thomas D.; Pfander, Claudia; Schwach, Frank; Brochet, Mathieu; Bushell, Ellen; Goulding, David; Sanders, Mandy; Lefebvre, Paul A.; Pei, Jimin; Grishin, Nick V.; Vanderlaan, Gary; Billker, Oliver; Snell, William J.

    2013-01-01

    Fertilization is a crucial yet poorly characterized event in eukaryotes. Our previous discovery that the broadly conserved protein HAP2 (GCS1) functioned in gamete membrane fusion in the unicellular green alga Chlamydomonas and the malaria pathogen Plasmodium led us to exploit the rare biological phenomenon of isogamy in Chlamydomonas in a comparative transcriptomics strategy to uncover additional conserved sexual reproduction genes. All previously identified Chlamydomonas fertilization-essential genes fell into related clusters based on their expression patterns. Out of several conserved genes in a minus gamete cluster, we focused on Cre06.g280600, an ortholog of the fertilization-related Arabidopsis GEX1. Gene disruption, cell biological, and immunolocalization studies show that CrGEX1 functions in nuclear fusion in Chlamydomonas. Moreover, CrGEX1 and its Plasmodium ortholog, PBANKA_113980, are essential for production of viable meiotic progeny in both organisms and thus for mosquito transmission of malaria. Remarkably, we discovered that the genes are members of a large, previously unrecognized family whose first-characterized member, KAR5, is essential for nuclear fusion during yeast sexual reproduction. Our comparative transcriptomics approach provides a new resource for studying sexual development and demonstrates that exploiting the data can lead to the discovery of novel biology that is conserved across distant taxa. PMID:23699412

  20. Comparative genomics in Chlamydomonas and Plasmodium identifies an ancient nuclear envelope protein family essential for sexual reproduction in protists, fungi, plants, and vertebrates.

    PubMed

    Ning, Jue; Otto, Thomas D; Pfander, Claudia; Schwach, Frank; Brochet, Mathieu; Bushell, Ellen; Goulding, David; Sanders, Mandy; Lefebvre, Paul A; Pei, Jimin; Grishin, Nick V; Vanderlaan, Gary; Billker, Oliver; Snell, William J

    2013-05-15

    Fertilization is a crucial yet poorly characterized event in eukaryotes. Our previous discovery that the broadly conserved protein HAP2 (GCS1) functioned in gamete membrane fusion in the unicellular green alga Chlamydomonas and the malaria pathogen Plasmodium led us to exploit the rare biological phenomenon of isogamy in Chlamydomonas in a comparative transcriptomics strategy to uncover additional conserved sexual reproduction genes. All previously identified Chlamydomonas fertilization-essential genes fell into related clusters based on their expression patterns. Out of several conserved genes in a minus gamete cluster, we focused on Cre06.g280600, an ortholog of the fertilization-related Arabidopsis GEX1. Gene disruption, cell biological, and immunolocalization studies show that CrGEX1 functions in nuclear fusion in Chlamydomonas. Moreover, CrGEX1 and its Plasmodium ortholog, PBANKA_113980, are essential for production of viable meiotic progeny in both organisms and thus for mosquito transmission of malaria. Remarkably, we discovered that the genes are members of a large, previously unrecognized family whose first-characterized member, KAR5, is essential for nuclear fusion during yeast sexual reproduction. Our comparative transcriptomics approach provides a new resource for studying sexual development and demonstrates that exploiting the data can lead to the discovery of novel biology that is conserved across distant taxa.

  1. On Preliminary Breakdown

    NASA Astrophysics Data System (ADS)

    Beasley, W. H.; Petersen, D.

    2013-12-01

    The preliminary breakdown phase of a negative cloud-to-ground lightning flash was observed in detail. Observations were made with a Photron SA1.1 high-speed video camera operating at 9,000 frames per second, fast optical sensors, a flat-plate electric field antenna covering the SLF to MF band, and VHF and UHF radio receivers with bandwidths of 20 MHz. Bright stepwise extensions of a negative leader were observed at an altitude of 8 km during the first few milliseconds of the flash, and were coincident with bipolar electric field pulses called 'characteristic pulses'. The 2-D step lengths of the preliminary processes were in excess of 100 meters, with some 2-D step lengths in excess of 200 meters. Smaller and shorter unipolar electric field pulses were superposed onto the bipolar electric field pulses, and were coincident with VHF and UHF radio pulses. After a few milliseconds, the emerging negative stepped leader system showed a marked decrease in luminosity, step length, and propagation velocity. Details of these events will be discussed, including the possibility that the preliminary breakdown phase consists not of a single developing lightning leader system, but of multiple smaller lightning leader systems that eventually join together into a single system.

  2. Safeguards Envelope Progress FY09

    SciTech Connect

    Richard Metcalf; Robert Bean

    2009-09-01

    The Safeguards Envelope is a strategy to determine a set of specific operating parameters which nuclear facilities may operate within to maximize safeguards effectiveness without sacrificing safety or plant efficiency. This paper details advanced statistical techniques will be applied to real plant process monitoring (PM) data from the Idaho Chemical Processing Plant (ICPP). As a result of the U.S. having no operating nuclear chemical reprocessing plants, there has been a strong interest in obtaining process monitoring data from the ICPP. The ICPP was shut down in 1996 and a recent effort has been made to retrieve the PM data from storage in a data mining effort. In a simulation based on this data, multi-tank and multi-attribute correlations were tested against synthetic diversion scenarios. Kernel regression smoothing was used to fit a curve to the historical data, and multivariable, residual analysis and cumulative sum techniques set parameters for operating conditions. Diversion scenarios were created and tested, showing improved results when compared with a previous study utilizing only one-variable Z- testing7.

  3. Breakdown of organic insulators

    NASA Technical Reports Server (NTRS)

    Cuddihy, E. F.

    1983-01-01

    Solar cells and their associated electrical interconnects and leads were encapsulated in transparent elastomeric materials. Their purpose in a photovoltaic module, one of the most important for these elastomeric encapsulation materials, is to function as electrical insulation. This includes internal insulation between adjacent solar cells, between other encapsulated electrical parts, and between the total internal electrical circuitry and external metal frames, grounded areas, and module surfaces. Catastrophic electrical breakdown of the encapsulant insulation materials or electrical current through these materials or module edges to external locations can lead to module failure and can create hazards to humans. Electrical insulation stability, advanced elastomeric encapsulation materials are developed which are intended to be intrinsically free of in-situ ionic impurities, have ultralow water absorption, be weather-stable (UV, oxygen), and have high mechanical flexibility. Efforts to develop a method of assessing the life potential of organic insulation materials in photovoltaic modules are described.

  4. Refrigerated cryogenic envelope

    DOEpatents

    Loudon, John D.

    1976-11-16

    An elongated cryogenic envelope including an outer tube and an inner tube coaxially spaced within said inner tube so that the space therebetween forms a vacuum chamber for holding a vacuum. The inner and outer tubes are provided with means for expanding or contracting during thermal changes. A shield is located in the vacuum chamber intermediate the inner and outer tubes; and, a refrigeration tube for directing refrigeration to the shield is coiled about at least a portion of the inner tube within the vacuum chamber to permit the refrigeration tube to expand or contract along its length during thermal changes within said vacuum chamber.

  5. Blueprint for Breakdown: Three Mile Island and the Media before the Accident.

    ERIC Educational Resources Information Center

    Friedman, Sharon M.

    1981-01-01

    Discusses media coverage of the Three Mile Island nuclear power plant before and during the disaster. Concludes that there was a communication breakdown prior to the accident. Outlines the causes and offers suggestions for avoiding similar breakdowns in the future. (JMF)

  6. High gradient RF breakdown studies

    NASA Astrophysics Data System (ADS)

    Laurent, Lisa Leanne

    Higher accelerating gradients are required by future demands for TeV electron linear colliders. With higher energy comes the challenge of handling stronger electromagnetic fields in the accelerator structures and in the microwave sources that supply the power. A limit on the maximum field gradient is imposed by rf electrical breakdown. Investigating methods to achieve higher gradients and to better understand the mechanisms involved in the rf breakdown process has been the focal point of this study. A systematic series of rf breakdown experiments have been conducted at Stanford Linear Accelerator Center utilizing a transmission cavity operating in the TM020 mode. A procedure was developed to examine the high gradient section of the cavity in an electron microscope. The results have revealed that breakdown asymmetry exists between opposing high gradient surfaces. During breakdown, a plasma formation is detected localized near the surface with no visible evidence of an arc traversing the gap. These findings support the theory that high frequency rf breakdown is a single surface phenomenon. Other results from this study have shown that breakdown can occur at relatively low voltages when surface irregularities exist and along grain boundaries. A series of steps have been developed through this study that have significantly reduced the number of breakdowns that occur along grain boundaries. Testing under various vacuum conditions (10-11--10 -5 Torr) have revealed that while the breakdown threshold remained the same, the field emitted current density increased by almost two orders of magnitude. This suggests that the total field emitted current density is not the critical parameter in the initiation of high frequency vacuum breakdown. In the course of this study, microparticles were carefully tracked before and after rf processing. The outcome of this research suggests that expensive cleanroom facilities may not offer any advantage over practicing good cleaning and

  7. Delaying vortex breakdown by waves

    NASA Astrophysics Data System (ADS)

    Yao, M. F.; Jiang, L. B.; Wu, J. Z.; Ma, H. Y.; Pan, J. Y.

    1989-03-01

    The effect of spiral waves on delaying vortex breakdown in a tube is studied experimentally and theoretically. When a harmonic oscillation was imposed on one of guiding vanes in the tube, the breakdown was observed to be postponed appreciately. According to the generalized Lagrangian mean theory, proper forcing spiral waves may produce an additional streaming momentum, of which the effect is favorable and similar to an axial suction at downstream end. The delayed breakdown position is further predicted by using nonlinear wave theory. Qualitative agreement between theory and experiment is obtained, and experimental comparison of the effects due to forcing spiral wave and axial suction is made.

  8. Breakdown in the pretext tokamak

    SciTech Connect

    Benesch, J.F.

    1981-06-01

    Data are presented on the application of ion cyclotron resonance RF power to preionization in tokamaks. We applied 0.3-3 kW at 12 MHz to hydrogen and obtained a visible discharge, but found no scaling of breakdown voltage with any parameter we were able to vary. A possible explanation for this, which implies that higher RF power would have been much more effective, is discussed. Finally, we present our investigation of the dV/dt dependence of breakdown voltage in PRETEXT, a phenomenon also seen in JFT-2. The breakdown is discussed in terms of the physics of Townsend discharges.

  9. RF breakdown experiments at SLAC

    NASA Astrophysics Data System (ADS)

    Laurent, L.; Scheitrum, G.; Vlieks, A.; Pearson, C.; Caryotakis, G.; Luhmann, N. C.

    1999-05-01

    RF breakdown is a critical issue in the conditioning of klystrons, accelerator sections, and rf components for the next linear collider (NLC), as well as other high gradient accelerators and high power microwave sources. SLAC is conducting a series of experiments using an X-band traveling wave ring to characterize the processes and trigger mechanisms associated with rf breakdown. The goal of the research is to identify materials, processes, and manufacturing methods that will increase the breakdown threshold and minimize the time required for conditioning.

  10. Nonlinear Theory and Breakdown

    NASA Technical Reports Server (NTRS)

    Smith, Frank

    2007-01-01

    The main points of recent theoretical and computational studies on boundary-layer transition and turbulence are to be highlighted. The work is based on high Reynolds numbers and attention is drawn to nonlinear interactions, breakdowns and scales. The research focuses in particular on truly nonlinear theories, i.e. those for which the mean-flow profile is completely altered from its original state. There appear to be three such theories dealing with unsteady nonlinear pressure-displacement interactions (I), with vortex/wave interactions (II), and with Euler-scale flows (III). Specific recent findings noted for these three, and in quantitative agreement with experiments, are the following. Nonlinear finite-time break-ups occur in I, leading to sublayer eruption and vortex formation; here the theory agrees with experiments (Nishioka) regarding the first spike. II gives rise to finite-distance blowup of displacement thickness, then interaction and break-up as above; this theory agrees with experiments (Klebanoff, Nishioka) on the formation of three-dimensional streets. III leads to the prediction of turbulent boundary-layer micro-scale, displacement-and stress-sublayer-thicknesses.

  11. Numerical simulation of vortex breakdown

    NASA Technical Reports Server (NTRS)

    Shi, X.

    1985-01-01

    The breakdown of an isolated axisymmetric vortex embedded in an unbounded uniform flow is examined by numerical integration of the complete Navier-Stokes equations for unsteady axisymmetric flow. Results show that if the vortex strength is small, the solution approaches a steady flow and the vortex is stable. If the strength is large enough, the solution remains unsteady and a recirculating zone will appear near the axis, its form and internal structure resembling those of the axisymmetric breakdown bubbles with multi-cells observed by Faler and Leibovich (1978). For apppropriate combinations of flow parameters, the flow reveals quasi-periodicity. Parallel calculations with the quasi-cylindrical approximation indicate that so far as predicting of breakdown is concerned, its results coincide quite well with the results mentioned above. Both show that the vortex breakdown has little concern with the Reynolds number or with the critical classification of the upstream flow, at least for the lower range of Reynolds numbers.

  12. Anisotropic charged core envelope star

    NASA Astrophysics Data System (ADS)

    Mafa Takisa, P.; Maharaj, S. D.

    2016-08-01

    We study a charged compact object with anisotropic pressures in a core envelope setting. The equation of state is quadratic in the core and linear in the envelope. There is smooth matching between the three regions: the core, envelope and the Reissner-Nordström exterior. We show that the presence of the electric field affects the masses, radii and compactification factors of stellar objects with values which are in agreement with previous studies. We investigate in particular the effect of electric field on the physical features of the pulsar PSR J1614-2230 in the core envelope model. The gravitational potentials and the matter variables are well behaved within the stellar object. We demonstrate that the radius of the core and the envelope can vary by changing the parameters in the speed of sound.

  13. Multifamily Envelope Leakage Model

    SciTech Connect

    Faakye, Omari; Griffiths, Dianne

    2015-05-08

    “The cost for blower testing is high, because it is labor intensive, and it may disrupt occupants in multiple units. This high cost and disruption deter program participants, and dissuade them from pursuing energy improvements that would trigger air leakage testing, such as improvements to the building envelope.” This statement found in a 2012 report by Heschong Mahone Group for several California interests emphasizes the importance of reducing the cost and complexity of blower testing in multifamily buildings. Energy efficiency opportunities are being bypassed. The cost of single blower testing is on the order of $300. The cost for guarded blower door testing—the more appropriate test for assessing energy savings opportunities—could easily be six times that, and that’s only if you have the equipment and simultaneous access to multiple apartments. Thus, the proper test is simply not performed. This research seeks to provide an algorithm for predicting the guarded blower door test result based upon a single, total blower door test.

  14. Dielectric breakdown of cell membranes.

    PubMed

    Zimmermann, U; Pilwat, G; Riemann, F

    1974-11-01

    With human and bovine red blood cells and Escherichia coli B, dielectric breakdown of cell membranes could be demonstrated using a Coulter Counter (AEG-Telefunken, Ulm, West Germany) with a hydrodynamic focusing orifice. In making measurements of the size distributions of red blood cells and bacteria versus increasing electric field strength and plotting the pulse heights versus the electric field strength, a sharp bend in the otherwise linear curve is observed due to the dielectric breakdown of the membranes. Solution of Laplace's equation for the electric field generated yields a value of about 1.6 V for the membrane potential at which dielectric breakdown occurs with modal volumes of red blood cells and bacteria. The same value is also calculated for red blood cells by applying the capacitor spring model of Crowley (1973. Biophys. J. 13:711). The corresponding electric field strength generated in the membrane at breakdown is of the order of 4 . 10(6) V/cm and, therefore, comparable with the breakdown voltages for bilayers of most oils. The critical detector voltage for breakdown depends on the volume of the cells. The volume-dependence predicted by Laplace theory with the assumption that the potential generated across the membrane is independent of volume, could be verified experimentally. Due to dielectric breakdown the red blood cells lose hemoglobin completely. This phenomenon was used to study dielectric breakdown of red blood cells in a homogeneous electric field between two flat platinum electrodes. The electric field was applied by discharging a high voltage storage capacitor via a spark gap. The calculated value of the membrane potential generated to produce dielectric breakdown in the homogeneous field is of the same order as found by means of the Coulter Counter. This indicates that mechanical rupture of the red blood cells by the hydrodynamic forces in the orifice of the Coulter Counter could also be excluded as a hemolysing mechanism. The detector

  15. Electrical Breakdown in Water Vapor

    SciTech Connect

    Skoro, N.; Maric, D.; Malovic, G.; Petrovic, Z. Lj.; Graham, W. G.

    2011-11-15

    In this paper investigations of the voltage required to break down water vapor are reported for the region around the Paschen minimum and to the left of it. In spite of numerous applications of discharges in biomedicine, and recent studies of discharges in water and vapor bubbles and discharges with liquid water electrodes, studies of the basic parameters of breakdown are lacking. Paschen curves have been measured by recording voltages and currents in the low-current Townsend regime and extrapolating them to zero current. The minimum electrical breakdown voltage for water vapor was found to be 480 V at a pressure times electrode distance (pd) value of around 0.6 Torr cm ({approx}0.8 Pa m). The present measurements are also interpreted using (and add additional insight into) the developing understanding of relevant atomic and particularly surface processes associated with electrical breakdown.

  16. Electrical breakdown in tissue electroporation.

    PubMed

    Guenther, Enric; Klein, Nina; Mikus, Paul; Stehling, Michael K; Rubinsky, Boris

    2015-11-27

    Electroporation, the permeabilization of the cell membrane by brief, high electric fields, has become an important technology in medicine for diverse application ranging from gene transfection to tissue ablation. There is ample anecdotal evidence that the clinical application of electroporation is often associated with loud sounds and extremely high currents that exceed the devices design limit after which the devices cease to function. The goal of this paper is to elucidate and quantify the biophysical and biochemical basis for this phenomenon. Using an experimental design that includes clinical data, a tissue phantom, sound, optical, ultrasound and MRI measurements, we show that the phenomenon is caused by electrical breakdown across ionized electrolysis produced gases near the electrodes. The breakdown occurs primarily near the cathode. Electrical breakdown during electroporation is a biophysical phenomenon of substantial importance to the outcome of clinical applications. It was ignored, until now.

  17. Measurement of breakdown current in dielectric materials

    NASA Astrophysics Data System (ADS)

    Pakhotin, V. A.; Zakrevskii, V. A.; Sudar', N. T.

    2015-08-01

    A new method to determine the resistance of the breakdown channel, current, and characteristic time is based on the measurements of the breakdown current pulse in a wide range of parameters of the measurement circuit. A problem with time-dependent resistance of the breakdown channel is numerically solved. An experimental variation in the resistance of the breakdown channel can be used to estimate the breakdown time. The method is tested with the aid of computer experiments and employed in the analysis of oscillograms of breakdown current in experiments with a dielectric polymer.

  18. Threshold criteria for undervoltage breakdown

    NASA Astrophysics Data System (ADS)

    Cooley, James E.; Choueiri, Edgar Y.

    2008-05-01

    The conditions under which an externally supplied pulse of electrons will induce breakdown in an undervoltaged, low-gain discharge gap are experimentally and theoretically explored. The minimum number of injected electrons required to achieve breakdown in a parallel-plate gap is measured in argon at pd values of 3-10 Torr m using ultraviolet laser pulses to photoelectrically release electrons from the cathode. This value was found to scale inversely with voltage at constant pd and with pressure within the parameter range explored. A dimensionless theoretical description of the phenomenon is formulated and numerically solved. It is found that a significant fraction of the charge on the plates must be injected for breakdown to be achieved at low gain. It is also found that fewer electrons are required as the gain due to electron-impact ionization (α process) is increased, or as the sensitivity of the α process to electric field is enhanced by increasing the gas pressure. A predicted insensitivity to ion mobility implies that the breakdown is determined during the first electron avalanche when space-charge distortion is greatest.

  19. The structure of vortex breakdown

    NASA Technical Reports Server (NTRS)

    Leibovich, S.

    1978-01-01

    The term 'vortex breakdown', as used in the reported investigation, refers to a disturbance characterized by the formation of an internal stagnation point on the vortex axis, followed by reversed flow in a region of limited axial extent. Two forms of vortex breakdown, which predominate, are shown in photographs. One form is called 'near-axisymmetric' (sometimes 'axisymmetric'), and the other is called 'spiral'. A survey is presented of work published since the 1972 review by Hall. Most experimental data taken since Hall's review have been in tubes, and the survey deals primarily with such cases. It is found that the assumption of axial-symmetry has produced useful results. The classification of flows as supercritical or subcritical, a step that assumes symmetry, has proved universally useful. Experiments show that vortex breakdown is always preceded by an upstream supercritical flow and followed by a subcritical wake. However, a comparison between experiments and attempts at prediction is less than encouraging. For a satisfactory understanding of the structure of vortex breakdown it is apparently necessary to take into account also aspects of asymmetry.

  20. Personnel occupied woven envelope robot

    NASA Technical Reports Server (NTRS)

    Wessling, F. C.

    1986-01-01

    The use of nonmetallic or fabric structures for space application is considered. The following structures are suggested: (1) unpressurized space hangars; (2) extendable tunnels for soft docking; and (3) manned habitat for space stations, storage facilities, and work structures. The uses of the tunnel as a passageway: for personnel and equipment, eliminating extravehicular activity, for access to a control cabin on a space crane and between free flyers and the space station are outlined. The personnal occupied woven envelope robot (POWER) device is shown. The woven envelope (tunnel) acts as part of the boom of a crane. Potential applications of POWER are outlined. Several possible deflection mechanisms and design criteria are determined.

  1. Carbon chemistry of circumstellar envelopes

    NASA Technical Reports Server (NTRS)

    Bieging, John H.

    1990-01-01

    The chemical composition of envelopes surrounding cool evolved stars, as determined from microwave spectroscopic observations, is reviewed. Emphasis is placed on recent observations with the new large mm-wavelength telescopes and interferometer arrays, and on new theoretical work, especially concerning ion-molecule chemistry of carbon-bearing in these envelopes. Thermal (as opposed to maser) emission lines are discussed. Much progress has been made in the past few years in the theoretical understanding of these objects. It is already clear, however, that observations with the new generation of mm-telescopes will require substantial improvements in the theoretical models to achieve a thorough understanding of the data now becoming available.

  2. Dielectric breakdown model for composite materials.

    PubMed

    Peruani, F; Solovey, G; Irurzun, I M; Mola, E E; Marzocca, A; Vicente, J L

    2003-06-01

    This paper addresses the problem of dielectric breakdown in composite materials. The dielectric breakdown model was generalized to describe dielectric breakdown patterns in conductor-loaded composites. Conducting particles are distributed at random in the insulating matrix, and the dielectric breakdown propagates according to new rules to take into account electrical properties and particle size. Dielectric breakdown patterns are characterized by their fractal dimension D and the parameters of the Weibull distribution. Studies are carried out as a function of the fraction of conducting inhomogeneities, p. The fractal dimension D of electrical trees approaches the fractal dimension of a percolation cluster when the fraction of conducting particles approximates the percolation limit. PMID:16241318

  3. Initiation of breakdown in slender compressible vortices

    NASA Technical Reports Server (NTRS)

    Krause, E.; Menne, S.; Liu, C. H.

    1986-01-01

    The initiation of the breakdown process for axially symmetric compressible flows is investigated using a numerical solution of the conservation equations for mass, momentum, and energy. The vortex is isolated, with its axis parallel to the direction of the main stream, and the core radius is small compared to the breakdown length. Computations for several flowfields indicate that the breakdown of the solution is shifted further downstream with increasing Mach number until breakdown is no longer observed. In the subsonic case, the influence of the initial temperature distribution on the breakdown length of the solution is more pronounced than in the supersonic case, with heating of the core enhancing breakdown, and cooling delaying it. The breakdown of the solution is seen to always occur for nonvanishing axial velocity components.

  4. Proposed RF Breakdown Studies at the AWA

    SciTech Connect

    Antipov, S.; Conde, M.; Gai, W.; Power, J.G.; Spentzouris, L.; Yusof, Z.; Dolgashev, V.; /SLAC

    2007-03-21

    A study of breakdown mechanism has been initiated at the Argonne Wakefield Accelerator (AWA). Breakdown may include several factors such as local field enhancement, explosive electron emission, Ohmic heating, tensile stress produced by electric field, and others. The AWA is building a dedicated facility to test various models for breakdown mechanisms and to determine the roles of different factors in the breakdown. We plan to trigger breakdown events with a high-powered laser at various wavelengths (IR to UV) to determine the role of explosive electron emission in the breakdown process. Another experimental idea follows from the recent work on a Schottky-enabled photoemission in an RF photoinjector [1] that allows us to determine in situ the field enhancement factor on a cathode surface. Monitoring the field enhancement factor before and after the breakdown can shed some light on a number of observations such as the crater formation process.

  5. Reconceptualization of the Budget Envelope.

    ERIC Educational Resources Information Center

    Jefferson, Anne L.

    This paper reconceptualizes the purposes of education's budget envelope. Citing numerous examples of how policymakers consider resource allocations apart from the main concerns of individual programs, the people reallocations affect, and education's most important programs, it suggests that policymakers and finance officers reemphasize program and…

  6. Misdelivery at the Nuclear Pore Complex—Stopping a Virus Dead in Its Tracks

    PubMed Central

    Flatt, Justin W.; Greber, Urs F.

    2015-01-01

    Many viruses deliver their genomes into the host cell’s nucleus before they replicate. While onco-retroviruses and papillomaviruses tether their genomes to host chromatin upon mitotic breakdown of the nuclear envelope, lentiviruses, such as human immunodeficiency virus, adenoviruses, herpesviruses, parvoviruses, influenza viruses, hepatitis B virus, polyomaviruses, and baculoviruses deliver their genomes into the nucleus of post-mitotic cells. This poses the significant challenge of slipping a DNA or RNA genome past the nuclear pore complex (NPC) embedded in the nuclear envelope. Quantitative fluorescence imaging is shedding new light on this process, with recent data implicating misdelivery of viral genomes at nuclear pores as a bottleneck to virus replication. Here, we infer NPC functions for nuclear import of viral genomes from cell biology experiments and explore potential causes of misdelivery, including improper virus docking at NPCs, incomplete translocation, virus-induced stress and innate immunity reactions. We conclude by discussing consequences of viral genome misdelivery for viruses and host cells, and lay out future questions to enhance our understanding of this phenomenon. Further studies into viral genome misdelivery may reveal unexpected aspects about NPC structure and function, as well as aid in developing strategies for controlling viral infections to improve human health. PMID:26226003

  7. Envelope dynamics of iron-core supernova models

    NASA Technical Reports Server (NTRS)

    Barkat, Z.; Wheeler, J. C.; Buchler, J.-R.; Rakavy, G.

    1974-01-01

    Wilson (1971) has found that the neutrino transport mechanism is unable to generate a supernova explosion in stars with collapsing iron cores. The present work uses Wilson's analysis to investigate the behavior of the overlying potentially explosive layers which Wilson omitted. The outer boundary of the core of Wilson's models moves in such a manner as to deliver a shock to the base of the envelope. We have numerically followed the progress of such shocks into the envelope of a realistic model obtained from evolutionary calculations. We find that only shocks so strong as to be inconsistent with our treatment are capable of ejecting material. For reasonable shocks the nuclear burning does not proceed rapidly at densities below 1,000 kg/cu cm, and the nuclear energy released is less than the shock energy in all models that come near to ejecting matter.

  8. Preparation of envelope membrane fractions from Arabidopsis chloroplasts for proteomic analysis and other studies.

    PubMed

    Salvi, Daniel; Moyet, Lucas; Seigneurin-Berny, Daphné; Ferro, Myriam; Joyard, Jacques; Rolland, Norbert

    2011-01-01

    Plastids are semiautonomous organelles restricted to plants and protists. These plastids are surrounded by a double membrane system, or envelope. These envelope membranes contain machineries to import nuclear-encoded proteins, and transporters for ions or metabolites, but are also essential for a range of plastid-specific metabolisms. Targeted semiquantitative proteomic investigations have revealed specific cross-contaminations by other cell or plastid compartments that may occur during chloroplast envelope purification. This article describes procedures developed to recover highly purified envelope fractions starting from Percoll-purified Arabidopsis chloroplasts, gives an overview of possible cross-contaminations, provides some tricks to limit these cross-contaminations, and lists immunological markers and methods that can be used to assess the purity of the envelope fractions.

  9. Time dependent breakdown in silicon dioxide films

    NASA Technical Reports Server (NTRS)

    Svensson, C.; Shumka, A.

    1975-01-01

    An investigation was conducted regarding the possible existence of a time-dependent breakdown mechanism in thermal oxides of the type used as gate oxide in MOS circuits. Questions of device fabrication are discussed along with details concerning breakdown measurements and the determination of C-V characteristics. A relatively large prebreakdown current observed in one of the cases is related to the time-dependent breakdown.

  10. Work Breakdown Structure (WBS) Handbook

    NASA Technical Reports Server (NTRS)

    2010-01-01

    The purpose of this document is to provide program/project teams necessary instruction and guidance in the best practices for Work Breakdown Structure (WBS) and WBS dictionary development and use for project implementation and management control. This handbook can be used for all types of NASA projects and work activities including research, development, construction, test and evaluation, and operations. The products of these work efforts may be hardware, software, data, or service elements (alone or in combination). The aim of this document is to assist project teams in the development of effective work breakdown structures that provide a framework of common reference for all project elements. The WBS and WBS dictionary are effective management processes for planning, organizing, and administering NASA programs and projects. The guidance contained in this document is applicable to both in-house, NASA-led effort and contracted effort. It assists management teams from both entities in fulfilling necessary responsibilities for successful accomplishment of project cost, schedule, and technical goals. Benefits resulting from the use of an effective WBS include, but are not limited to: providing a basis for assigned project responsibilities, providing a basis for project schedule development, simplifying a project by dividing the total work scope into manageable units, and providing a common reference for all project communication.

  11. Relativistic breakdown in planetary atmospheres

    SciTech Connect

    Dwyer, J. R.

    2007-04-15

    In 2003, a new electrical breakdown mechanism involving the production of runaway avalanches by positive feedback from runaway positrons and energetic photons was introduced. This mechanism, which shall be referred to as 'relativistic feedback', allows runaway discharges in gases to become self-sustaining, dramatically increasing the flux of runaway electrons, the accompanying high-energy radiation, and resulting ionization. Using detailed Monte Carlo calculations, properties of relativistic feedback are investigated. It is found that once relativistic feedback fully commences, electrical breakdown will occur and the ambient electric field, extending over cubic kilometers, will be discharged in as little as 2x10{sup -5} s. Furthermore, it is found that the flux of energetic electrons and x rays generated by this mechanism can exceed the flux generated by the standard relativistic runaway electron model by a factor of 10{sup 13}, making relativistic feedback a good candidate for explaining terrestrial gamma-ray flashes and other high-energy phenomena observed in the Earth's atmosphere.

  12. Safeguards Envelope Progress FY08

    SciTech Connect

    Robert Bean; Richard Metcalf; Aaron Bevill

    2008-09-01

    The Safeguards Envelope Project met its milestones by creating a rudimentary safeguards envelope, proving the value of the approach on a small scale, and determining the most appropriate path forward. The Idaho Chemical Processing Plant’s large cache of reprocessing process monitoring data, dubbed UBER Data, was recovered and used in the analysis. A probabilistic Z test was used on a Markov Monte Carlo simulation of expected diversion data when compared with normal operating data. The data regarding a fully transient event in a tank was used to create a simple requirement, representative of a safeguards envelope, whose impact was a decrease in operating efficiency by 1.3% but an increase in material balance period of 26%. This approach is operator, state, and international safeguards friendly and should be applied to future reprocessing plants. Future requirements include tank-to-tank correlations in reprocessing facilities, detailed operations impact studies, simulation inclusion, automated optimization, advanced statistics analysis, and multi-attribute utility analysis.

  13. Transparent ceramic lamp envelope materials

    NASA Astrophysics Data System (ADS)

    Wei, G. C.

    2005-09-01

    Transparent ceramic materials with optical qualities comparable to single crystals of similar compositions have been developed in recent years, as a result of the improved understanding of powder-processing-fabrication- sintering-property inter-relationships. These high-temperature materials with a range of thermal and mechanical properties are candidate envelopes for focused-beam, short-arc lamps containing various fills operating at temperatures higher than quartz. This paper reviews the composition, structure and properties of transparent ceramic lamp envelope materials including sapphire, small-grained polycrystalline alumina, aluminium oxynitride, yttrium aluminate garnet, magnesium aluminate spinel and yttria-lanthana. A satisfactory thermal shock resistance is required for the ceramic tube to withstand the rapid heating and cooling cycles encountered in lamps. Thermophysical properties, along with the geometry, size and thickness of a transparent ceramic tube, are important parameters in the assessment of its resistance to fracture arising from thermal stresses in lamps during service. The corrosive nature of lamp-fill liquid and vapour at high temperatures requires that all lamp components be carefully chosen to meet the target life. The wide range of new transparent ceramics represents flexibility in pushing the limit of envelope materials for improved beamer lamps.

  14. Excitation of gravity waves in common envelopes

    NASA Technical Reports Server (NTRS)

    Soker, Noam

    1992-01-01

    We study the excitation of gravity waves by a low-mass companion orbiting inside the envelope of a giant star, concentrating on brown dwarfs inside the envelope of asymptotic giant branch stars. Efficient g-wave excitations occur only after the brown dwarf has spiraled-in to the radiative zone, well inside the envelope, of the asymptotic giant branch star. The brown dwarf excites g-waves when its orbital radius is about 3-10 solar radii. At this stage of the evolution the envelope mass is below 0.1 solar mass. The g-waves propagate inward from the secondary orbit, carrying angular momentum and energy. We find that the angular momentum transport leads to an efficient spin-up of the inner envelopes. The differential rotation between the envelope and core and nonlinear wave effects, can cause a mixing of heavy elements from the core to the envelope.

  15. Thermal dielectric breakdown with cylindrical electrodes

    NASA Technical Reports Server (NTRS)

    Beers, B. L.; Odwyer, J. J.

    1983-01-01

    Solutions to the equations of thermal breakdown are computed for cylindrial electrodes with different boundary conditions. The development of the electric field and the temperature distributions are followed as functions of time, consequent on the application of a constant interelectrode voltage. The implications for possible breakdown by modes other than thermal are briefly discussed.

  16. Fundamental studies on passivity and passivity breakdown

    SciTech Connect

    Macdonald, D.D.; Urquidi-Macdonald, M.

    1993-06-01

    Using photoelectrochemical impedance and admittance spectroscopies, a fundamental and quantitative understanding of the mechanisms for the growth and breakdown of passive films on metal and alloy surfaces in contact with aqueous environments is being developed. A point defect model has been extended to explain the breakdown of passive films, leading to pitting and crack growth and thus development of damage due to localized corrosion.

  17. Breakdown of interdependent directed networks.

    PubMed

    Liu, Xueming; Stanley, H Eugene; Gao, Jianxi

    2016-02-01

    Increasing evidence shows that real-world systems interact with one another via dependency connectivities. Failing connectivities are the mechanism behind the breakdown of interacting complex systems, e.g., blackouts caused by the interdependence of power grids and communication networks. Previous research analyzing the robustness of interdependent networks has been limited to undirected networks. However, most real-world networks are directed, their in-degrees and out-degrees may be correlated, and they are often coupled to one another as interdependent directed networks. To understand the breakdown and robustness of interdependent directed networks, we develop a theoretical framework based on generating functions and percolation theory. We find that for interdependent Erdős-Rényi networks the directionality within each network increases their vulnerability and exhibits hybrid phase transitions. We also find that the percolation behavior of interdependent directed scale-free networks with and without degree correlations is so complex that two criteria are needed to quantify and compare their robustness: the percolation threshold and the integrated size of the giant component during an entire attack process. Interestingly, we find that the in-degree and out-degree correlations in each network layer increase the robustness of interdependent degree heterogeneous networks that most real networks are, but decrease the robustness of interdependent networks with homogeneous degree distribution and with strong coupling strengths. Moreover, by applying our theoretical analysis to real interdependent international trade networks, we find that the robustness of these real-world systems increases with the in-degree and out-degree correlations, confirming our theoretical analysis. PMID:26787907

  18. Breakdown of interdependent directed networks.

    PubMed

    Liu, Xueming; Stanley, H Eugene; Gao, Jianxi

    2016-02-01

    Increasing evidence shows that real-world systems interact with one another via dependency connectivities. Failing connectivities are the mechanism behind the breakdown of interacting complex systems, e.g., blackouts caused by the interdependence of power grids and communication networks. Previous research analyzing the robustness of interdependent networks has been limited to undirected networks. However, most real-world networks are directed, their in-degrees and out-degrees may be correlated, and they are often coupled to one another as interdependent directed networks. To understand the breakdown and robustness of interdependent directed networks, we develop a theoretical framework based on generating functions and percolation theory. We find that for interdependent Erdős-Rényi networks the directionality within each network increases their vulnerability and exhibits hybrid phase transitions. We also find that the percolation behavior of interdependent directed scale-free networks with and without degree correlations is so complex that two criteria are needed to quantify and compare their robustness: the percolation threshold and the integrated size of the giant component during an entire attack process. Interestingly, we find that the in-degree and out-degree correlations in each network layer increase the robustness of interdependent degree heterogeneous networks that most real networks are, but decrease the robustness of interdependent networks with homogeneous degree distribution and with strong coupling strengths. Moreover, by applying our theoretical analysis to real interdependent international trade networks, we find that the robustness of these real-world systems increases with the in-degree and out-degree correlations, confirming our theoretical analysis.

  19. Breakdown properties of irradiated MOS capacitors

    SciTech Connect

    Paccagnella, A.; Candelori, A. |; Milani, A.; Formigoni, E.; Ghidini, G.; Drera, D.; Pellizzer, F. |; Fuochi, P.G.; Lavale, M.

    1996-12-01

    The authors have studied the effects of ionizing and non-ionizing radiation on the breakdown properties of different types of MOS capacitors, with thick (200 nm) and thin (down to 8 nm) oxides. In general, no large variations of the average breakdown field, time-to-breakdown at constant voltage, or charge-to-breakdown at constant voltage, or charge-to-breakdown values have been observed after high dose irradiation (20 Mrad(Si) 9 MeV electrons on thin and thick oxides, 17(Si) Mrad Co{sup 60} gamma and 10{sup 14} neutrons/cm{sup 2} only on thick oxides). However, some modifications of the cumulative failure distributions have been observed in few of the oxides tested.

  20. Cell entry of enveloped viruses.

    PubMed

    Cosset, François-Loic; Lavillette, Dimitri

    2011-01-01

    Enveloped viruses penetrate their cell targets following the merging of their membrane with that of the cell. This fusion process is catalyzed by one or several viral glycoproteins incorporated on the membrane of the virus. These envelope glycoproteins (EnvGP) evolved in order to combine two features. First, they acquired a domain to bind to a specific cellular protein, named "receptor." Second, they developed, with the help of cellular proteins, a function of finely controlled fusion to optimize the replication and preserve the integrity of the cell, specific to the genus of the virus. Following the activation of the EnvGP either by binding to their receptors and/or sometimes the acid pH of the endosomes, many changes of conformation permit ultimately the action of a specific hydrophobic domain, the fusion peptide, which destabilizes the cell membrane and leads to the opening of the lipidic membrane. The comprehension of these mechanisms is essential to develop medicines of the therapeutic class of entry inhibitor like enfuvirtide (Fuzeon) against human immunodeficiency virus (HIV). In this chapter, we will summarize the different envelope glycoprotein structures that viruses develop to achieve membrane fusion and the entry of the virus. We will describe the different entry pathways and cellular proteins that viruses have subverted to allow infection of the cell and the receptors that are used. Finally, we will illustrate more precisely the recent discoveries that have been made within the field of the entry process, with a focus on the use of pseudoparticles. These pseudoparticles are suitable for high-throughput screenings that help in the development of natural or artificial inhibitors as new therapeutics of the class of entry inhibitors.

  1. Flexible Envelope Request Notation (FERN)

    NASA Technical Reports Server (NTRS)

    Zoch, David R.; Lavallee, David; Weinstein, Stuart

    1991-01-01

    The following topics are presented in view graph form and include the following: scheduling application; the motivation for the Flexible Envelope Request Notation (FERN); characteristics of FERN; types of information needed in requests; where information is stored in requests; FERN structures; generic requests; resource availability for pooled resources; expressive notation; temporal constraints; time formats; changes to FERN; sample FERN requests; the temporal relationship between two steps; maximum activity length to limit step delays; alternative requests; the temporal relationship between two activities; and idle resource usage between steps.

  2. Laser-Induced Breakdown Spectroscopy (LIBS): specific applications

    NASA Astrophysics Data System (ADS)

    Trtica, M. S.; Savovic, J.; Stoiljkovic, M.; Kuzmanovic, M.; Momcilovic, M.; Ciganovic, J.; Zivkovic, S.

    2015-12-01

    A short overview of Laser Induced Breakdown Spectroscopy (LIBS) with emphasis on the new trends is presented. Nowadays, due to unique features of this technique, LIBS has found applications in a great variety of fields. Achievements in the application of LIBS in nuclear area, for hazardous materials detection and in geology were considered. Also, some results recently obtained at VINCA Institute, with LIBS system based on transversely excited atmospheric (TEA) CO2 laser, are presented. Future investigations of LIBS will be oriented toward further improvement of the analytical performance of this technique, as well as on finding new application fields.

  3. Structural basis for membrane anchoring of HIV-1 envelope spike.

    PubMed

    Dev, Jyoti; Park, Donghyun; Fu, Qingshan; Chen, Jia; Ha, Heather Jiwon; Ghantous, Fadi; Herrmann, Tobias; Chang, Weiting; Liu, Zhijun; Frey, Gary; Seaman, Michael S; Chen, Bing; Chou, James J

    2016-07-01

    HIV-1 envelope spike (Env) is a type I membrane protein that mediates viral entry. We used nuclear magnetic resonance to determine an atomic structure of the transmembrane (TM) domain of HIV-1 Env reconstituted in bicelles that mimic a lipid bilayer. The TM forms a well-ordered trimer that protects a conserved membrane-embedded arginine. An amino-terminal coiled-coil and a carboxyl-terminal hydrophilic core stabilize the trimer. Individual mutations of conserved residues did not disrupt the TM trimer and minimally affected membrane fusion and infectivity. Major changes in the hydrophilic core, however, altered the antibody sensitivity of Env. These results show how a TM domain anchors, stabilizes, and modulates a viral envelope spike and suggest that its influence on Env conformation is an important consideration for HIV-1 immunogen design. PMID:27338706

  4. Circumplanetary disc or circumplanetary envelope?

    NASA Astrophysics Data System (ADS)

    Szulágyi, J.; Masset, F.; Lega, E.; Crida, A.; Morbidelli, A.; Guillot, T.

    2016-08-01

    We present three-dimensional simulations with nested meshes of the dynamics of the gas around a Jupiter mass planet with the JUPITER and FARGOCA codes. We implemented a radiative transfer module into the JUPITER code to account for realistic heating and cooling of the gas. We focus on the circumplanetary gas flow, determining its characteristics at very high resolution (80 per cent of Jupiter's diameter). In our nominal simulation where the temperature evolves freely by the radiative module and reaches 13000 K at the planet, a circumplanetary envelope was formed filling the entire Roche lobe. Because of our equation of state is simplified and probably overestimates the temperature, we also performed simulations with limited maximal temperatures in the planet region (1000, 1500, and 2000 K). In these fixed temperature cases circumplanetary discs (CPDs) were formed. This suggests that the capability to form a CPD is not simply linked to the mass of the planet and its ability to open a gap. Instead, the gas temperature at the planet's location, which depends on its accretion history, plays also fundamental role. The CPDs in the simulations are hot and cooling very slowly, they have very steep temperature and density profiles, and are strongly sub-Keplerian. Moreover, the CPDs are fed by a strong vertical influx, which shocks on the CPD surfaces creating a hot and luminous shock-front. In contrast, the pressure supported circumplanetary envelope is characterized by internal convection and almost stalled rotation.

  5. Phospholipid turnover and ultrastructural correlates during spontaneous germinal vesicle breakdown of the bovine oocyte: Effects of a cyclic AMP phosphodiesterase inhibitor

    SciTech Connect

    Homa, S.T.; Webster, S.D.; Russell, R.K. )

    1991-08-01

    The turnover of (32P)orthophosphate in bovine oocyte phospholipids was studied during the early stages of spontaneous meiotic maturation, and during inhibition of this process by the cAMP phosphodiesterase inhibitor 3-isobutyl-1-methyl-xanthine (IBMX). Radioactive lipids were separated by TLC and the meiotic stage was determined cytogenetically. Ultrastructure of the nuclear membrane was examined using transmission EM. During the commitment period to meiotic resumption, which precedes germinal vesicle breakdown (GVBD), small localized convolutions appeared in the intact nuclear membrane. This was accompanied by a decrease in (32P)phosphatidic acid (PA) and an increase in (32P)-phosphatidylcholine (PC). This was followed by extensive convolutions, and subsequent dissociation, of the nuclear membrane, concomitant with a tremendous surge in (32P)PC and (32P)phosphatidylethanolamine (PE). The cAMP-mediated maintenance of meiotic arrest involved retention of entire nuclear envelope integrity and total inhibition of the surge in (32P)PC and (32P)PE which accompanied GVBD. The increase in (32P)phosphatidylinositol (PI) associated with all stages of early meiotic resumption was unaffected by IBMX. Microinjection of heparin inhibited GVBD, and injection of inositol 1,4,5-trisphosphate (IP3) overrode IBMX-maintained meiotic arrest in almost 40% of the oocytes. The results suggest that there may be several functions for phospholipid turnover in the regulation of spontaneous meiotic resumption in the bovine oocyte. The first precedes the commitment period, and involves IP3 generation to serve as the primary signal for meiotic resumption. The second occurs concomitant with the commitment period, is unaffected by the level of intracellular cAMP, and is associated with the general turnover of phospholipid.

  6. Scintillation Breakdowns in Chip Tantalum Capacitors

    NASA Technical Reports Server (NTRS)

    Teverovsky, Alexander

    2008-01-01

    Scintillations in solid tantalum capacitors are momentarily local breakdowns terminated by a self-healing or conversion to a high-resistive state of the manganese oxide cathode. This conversion effectively caps the defective area of the tantalum pentoxide dielectric and prevents short-circuit failures. Typically, this type of breakdown has no immediate catastrophic consequences and is often considered as nuisance rather than a failure. Scintillation breakdowns likely do not affect failures of parts under surge current conditions, and so-called "proofing" of tantalum chip capacitors, which is a controllable exposure of the part after soldering to voltages slightly higher than the operating voltage to verify that possible scintillations are self-healed, has been shown to improve the quality of the parts. However, no in-depth studies of the effect of scintillations on reliability of tantalum capacitors have been performed so far. KEMET is using scintillation breakdown testing as a tool for assessing process improvements and to compare quality of different manufacturing lots. Nevertheless, the relationship between failures and scintillation breakdowns is not clear, and this test is not considered as suitable for lot acceptance testing. In this work, scintillation breakdowns in different military-graded and commercial tantalum capacitors were characterized and related to the rated voltages and to life test failures. A model for assessment of times to failure, based on distributions of breakdown voltages, and accelerating factors of life testing are discussed.

  7. Breakdown characteristics of xenon HID Lamps

    NASA Astrophysics Data System (ADS)

    Babaeva, Natalia; Sato, Ayumu; Brates, Nanu; Noro, Koji; Kushner, Mark

    2009-10-01

    The breakdown characteristics of mercury free xenon high intensity discharge (HID) lamps exhibit a large statistical time lag often having a large scatter in breakdown voltages. In this paper, we report on results from a computational investigation of the processes which determine the ignition voltages for positive and negative pulses in commercial HID lamps having fill pressures of up to 20 atm. Steep voltage rise results in higher avalanche electron densities and earlier breakdown times. Circuit characteristics also play a role. Large ballast resistors may limit current to the degree that breakdown is quenched. The breakdown voltage critically depends on cathode charge injection by electric field emission (or other mechanisms) which in large part controls the statistical time lag for breakdown. For symmetric lamps, ionization waves (IWs) simultaneously develop from the bottom and top electrodes. Breakdown typically occurs when the top and bottom IWs converge. Condensed salt layers having small conductivities on the inner walls of HID lamps and on the electrodes can influence the ignition behavior. With these layers, IWs tend to propagate along the inner wall and exhibit a different structure depending on the polarity.

  8. Nonstationary envelope process and first excursion probability.

    NASA Technical Reports Server (NTRS)

    Yang, J.-N.

    1972-01-01

    The definition of stationary random envelope proposed by Cramer and Leadbetter, is extended to the envelope of nonstationary random process possessing evolutionary power spectral densities. The density function, the joint density function, the moment function, and the crossing rate of a level of the nonstationary envelope process are derived. Based on the envelope statistics, approximate solutions to the first excursion probability of nonstationary random processes are obtained. In particular, applications of the first excursion probability to the earthquake engineering problems are demonstrated in detail.

  9. Fundamental studies of passivity and passivity breakdown

    SciTech Connect

    Macdonald, D.D.; Urquidi-Macdonald, M.; Song, H.; Biaggio-Rocha, S.; Searson, P.

    1991-11-01

    This report summarizes the findings of our fundamental research program on passivity and passivity breakdown. During the past three and one half years in this program (including the three year incrementally-funded grant prior to the present grant), we developed and experimentally tested various physical models for the growth and breakdown of passive films on metal surfaces. These models belong to a general class termed point defects models'' (PDMs), in which the growth and breakdown of passive films are described in terms of the movement of anion and cation vacancies.

  10. Electrical Breakdown of Submillimeter Water Gaps

    NASA Astrophysics Data System (ADS)

    Schoenbach, K. H.; Cooper, J.; Garner, A.; Goan, B.; Joshi, R. P.; Kolb, J.; Katsuki, S.; Kono, S.; Laroussi, M.; Leipold, F.; Lu, X.; Mallot, C.; Quian, J.; Xiao, S.

    2002-12-01

    Electrical breakdown and recovery processes in water have been studied using electrical and nanosecond optical diagnostics. The breakdown electric field in submillimeter gaps with 200 ns voltage pulses applied has been measured as 1 MV/cm, the rate of current rise during breakdown reaches 4ṡ1011 A/s. The switch recovery time is determined by expansion and decay of a vapor bubble. The experimental results, together with the results of a model with a percolative approach, provide design criteria for compact, high power, high repetition rate, liquid-switch pulse generators.

  11. Microwave gas breakdown in elliptical waveguides

    SciTech Connect

    Koufogiannis, I. D.; Sorolla, E. Mattes, M.

    2014-01-15

    This paper analyzes the microwave gas discharge within elliptical waveguides excited by the fundamental mode. The Rayleigh-Ritz method has been applied to solve the continuity equation. The eigenvalue problem defined by the breakdown condition has been solved and the effective diffusion length of the elliptical waveguide has been calculated, what is used to find the corona threshold. This paper extends the microwave breakdown model developed for circular waveguides and shows the better corona withstanding capabilities of elliptical waveguides. The corona breakdown electric field threshold obtained with the variational method has been compared with the one calculated with the Finite Elements Method, showing excellent agreement.

  12. [Fundamental studies of passivity and passivity breakdown

    SciTech Connect

    Macdonald, D.D.

    1993-07-01

    We developed and experimentally tested physical models for growth and breakdown of passive films on metal surfaces. These models are ``point defect models,`` in which the growth and breakdown are described in terms of movement of anion and cation vacancies. The work during the past 5 years resulted in: theory of growth and breakdown of passive films, theory of corrosion-resistant alloys, electronic structure of passive films, and estimation of damage functions for energy systems. Proposals are give for the five ongoing tasks. 10 figs.

  13. Early Site Permit Demonstration Program: Plant parameters envelope report. Volume 1

    SciTech Connect

    Not Available

    1993-03-01

    The Early Site Permit (ESP) Demonstration Program is the nuclear industry`s initiative for piloting the early resolution of siting-related issues before the detailed design proceedings of the combined operating license review. The ESP Demonstration Program consists of three phases. The plant parameters envelopes task is part of Phase 1, which addresses the generic review of applicable federal regulations and develops criteria for safety and environmental assessment of potential sites. The plant parameters envelopes identify parameters that characterize the interface between an ALWR design and a potential site, and quantify the interface through values selected from the Utility Requirements Documents, vendor design information, or engineering assessments. When augmented with site-specific information, the plant parameters envelopes provide sufficient information to allow ESPs to be granted based on individual ALWR design information or enveloping design information for the evolutionary, passive, or generic ALWR plants. This document is expected to become a living document when used by future applicants.

  14. Microwave air breakdown enhanced with metallic initiators

    SciTech Connect

    Herring, G. C.; Popovic, S.

    2008-03-31

    We have determined X-band (9.4 GHz) electric field strengths required to obtain air breakdown at atmospheric pressure in the presence of metallic initiators, which are irradiated with repetitive (30 pulses/s) microwave pulses of 3 {mu}s duration and 200 kW peak power. Using a half-wavelength initiator, a factor of 40 reduction (compared to no initiator) was observed in the electric field required to achieve breakdown. The present measurements are compared to a previously published model for air breakdown, which was originally validated with S-band (3 GHz) frequencies and single 40 {mu}s pulses. We find good agreement between this previous model and our present measurements of breakdown with X-band frequencies and repetitive 3 {mu}s pulses.

  15. Contributions to theory of vortex breakdown

    NASA Astrophysics Data System (ADS)

    Shivamoggi, B. K.; Uberoi, M. S.

    A study is made of vortex breakdown in stratified flows, and it is found that a positive stratification in the vortex where the density is increasing away from the axis, postpones the vortex breakdown and vice versa. This is apparent due to the density increasing in a direction opposite to that of an effective gravity which would correspond to a topheavy arrangement under gravity. It is also shown that a wavemotion promotes the possibility of axisymmetric flow downstream of the transaction.

  16. Initiation of breakdown in slender compressible vortices

    NASA Technical Reports Server (NTRS)

    Krause, E.; Menne, S.; Liu, C. H.

    1986-01-01

    The onset of vortex breakdown in compressible flows is investigated analytically for the case in which the flow is axially symmetric, the vortex is isolated, its axis is parallel to the main flow, and the vortex radius is small compared to the breakdown length. The conservation equations for mass, momentum, and energy are formulated and solved numerically using a finite-difference scheme, as described by Krause (1985); numerical results are presented in graphs and briefly characterized.

  17. Spark gap with low breakdown voltage jitter

    DOEpatents

    Rohwein, G.J.; Roose, L.D.

    1996-04-23

    Novel spark gap devices and electrodes are disclosed. The novel spark gap devices and electrodes are suitable for use in a variety of spark gap device applications. The shape of the electrodes gives rise to local field enhancements and reduces breakdown voltage jitter. Breakdown voltage jitter of approximately 5% has been measured in spark gaps according the invention. Novel electrode geometries and materials are disclosed. 13 figs.

  18. Spark gap with low breakdown voltage jitter

    DOEpatents

    Rohwein, Gerald J.; Roose, Lars D.

    1996-01-01

    Novel spark gap devices and electrodes are disclosed. The novel spark gap devices and electrodes are suitable for use in a variety of spark gap device applications. The shape of the electrodes gives rise to local field enhancements and reduces breakdown voltage jitter. Breakdown voltage jitter of approximately 5% has been measured in spark gaps according the invention. Novel electrode geometries and materials are disclosed.

  19. Surface breakdown igniter for mercury arc devices

    DOEpatents

    Bayless, John R.

    1977-01-01

    Surface breakdown igniter comprises a semiconductor of medium resistivity which has the arc device cathode as one electrode and has an igniter anode electrode so that when voltage is applied between the electrodes a spark is generated when electrical breakdown occurs over the surface of the semiconductor. The geometry of the igniter anode and cathode electrodes causes the igniter discharge to be forced away from the semiconductor surface.

  20. Electromechanics and Electrical Breakdown of Particulate Layers

    NASA Astrophysics Data System (ADS)

    Moslehi, Bizhan G. R.

    A comprehensive theory of the electromechanics and electrical breakdown of a current-carrying particulate layer is developed, which takes into account its inhomogeneous nature and mode of compaction. The theory treates the general case of combined surface and volume conduction and takes account of self-compression of the layer due to electrical forces. The electromechanical theory predicts the existence of a remarkably large electrical cohesive stress in the layer due to a strong field enhancement in and around the contact regions. Furthermore, it shows a decrease in the apparent resistivity of the layer with increasing electric field as a result of self-compression. The analysis of electrical breakdown of current -carrying particulate layer predicts the onset of breakdown of the layer in the form of intermittent microsparks in the gap between the contacting particles when the electric field at the contact or in the surrounding gap exceeds the threshold breakdown value. An analysis of the behavior of the layer after breakdown in terms of a simplified equivalent lumped circuit predicts increases of sparking frequency and average current as the applied average field exceeds the threshold average field for the onset of breakdown. The results of measurements on layers of glass beads and fly-ash in a standard resistivity cell are in good agreement with the theoretical predictions for the field-dependent resistivity characteristics. The work has particular significance for electrostatic precipitation and addresses the phenomenon of backdischarge and the questions of the retention, rapping, and reentrainment of precipitation ash layers.

  1. Humidity effects on wire insulation breakdown strength.

    SciTech Connect

    Appelhans, Leah

    2013-08-01

    Methods for the testing of the dielectric breakdown strength of insulation on metal wires under variable humidity conditions were developed. Two methods, an ASTM method and the twisted pair method, were compared to determine if the twisted pair method could be used for determination of breakdown strength under variable humidity conditions. It was concluded that, although there were small differences in outcomes between the two testing methods, the non-standard method (twisted pair) would be appropriate to use for further testing of the effects of humidity on breakdown performance. The dielectric breakdown strength of 34G copper wire insulated with double layer Poly-Thermaleze/Polyamide-imide insulation was measured using the twisted pair method under a variety of relative humidity (RH) conditions and exposure times. Humidity at 50% RH and below was not found to affect the dielectric breakdown strength. At 80% RH the dielectric breakdown strength was significantly diminished. No effect for exposure time up to 140 hours was observed at 50 or 80%RH.

  2. A computational study of the topology of vortex breakdown

    NASA Technical Reports Server (NTRS)

    Spall, Robert E.; Gatski, Thomas B.

    1991-01-01

    A fully three-dimensional numerical simulation of vortex breakdown using the unsteady, incompressible Navier-Stokes equations has been performed. Solutions to four distinct types of breakdown are identified and compared with experimental results. The computed solutions include weak helical, double helix, spiral, and bubble-type breakdowns. The topological structure of the various breakdowns as well as their interrelationship are studied. The data reveal that the asymmetric modes of breakdown may be subject to additional breakdowns as the vortex core evolves in the streamwise direction. The solutions also show that the freestream axial velocity distribution has a significant effect on the position and type of vortex breakdown.

  3. Dielectric breakdown in mineral oil ITO 100 based magnetic fluid

    NASA Astrophysics Data System (ADS)

    Kudelcik, J.; Bury, P.; Kopcansky, P.; Timko, M.

    The development of dielectric breakdown and the DC dielectric breakdown voltage of magnetic fluids based on inhibited transformer oil ITO 100 were investigated in parallel orientations of external magnetic field. It was shown that the breakdown voltage is strongly influenced by the magnetic nanoparticles. The magnetic fluids with the volume concentration 1and 0.2% had better dielectric properties than pure transformer oil. The increase of breakdown voltage was interpreted on the base of the bubble theory of breakdown.

  4. The Metabolite Transporters of the Plastid Envelope: An Update

    PubMed Central

    Facchinelli, Fabio; Weber, Andreas P. M.

    2011-01-01

    The engulfment of a photoautotrophic cyanobacterium by a primitive mitochondria-bearing eukaryote traces back to more than 1.2 billion years ago. This single endosymbiotic event not only provided the early petroalgae with the metabolic capacity to perform oxygenic photosynthesis, but also introduced a plethora of other metabolic routes ranging from fatty acids and amino acids biosynthesis, nitrogen and sulfur assimilation to secondary compounds synthesis. This implicated the integration and coordination of the newly acquired metabolic entity with the host metabolism. The interface between the host cytosol and the plastidic stroma became of crucial importance in sorting precursors and products between the plastid and other cellular compartments. The plastid envelope membranes fulfill different tasks: they perform important metabolic functions, as they are involved in the synthesis of carotenoids, chlorophylls, and galactolipids. In addition, since most genes of cyanobacterial origin have been transferred to the nucleus, plastidial proteins encoded by nuclear genes are post-translationally transported across the envelopes through the TIC–TOC import machinery. Most importantly, chloroplasts supply the photoautotrophic cell with photosynthates in form of reduced carbon. The innermost bilayer of the plastidic envelope represents the permeability barrier for the metabolites involved in the carbon cycle and is literally stuffed with transporter proteins facilitating their transfer. The intracellular metabolite transporters consist of polytopic proteins containing membrane spans usually in the number of four or more α-helices. Phylogenetic analyses revealed that connecting the plastid with the host metabolism was mainly a process driven by the host cell. In Arabidopsis, 58% of the metabolite transporters are of host origin, whereas only 12% are attributable to the cyanobacterial endosymbiont. This review focuses on the metabolite transporters of the inner envelope

  5. Personnel occupied woven envelope robot

    NASA Technical Reports Server (NTRS)

    Wessling, Francis; Teoh, William; Ziemke, M. Carl

    1988-01-01

    The Personnel Occupied Woven Envelope Robot (POWER) provides an alternative to extravehicular activity (EVA) of space suited astronauts and/or use of long slender manipulator arms such as are used in the Shuttle Remote Manipulator System. POWER provides the capability for a shirt sleeved astronaut to perform such work by entering a control pod through air locks at both ends of an inflated flexible bellows (access tunnel). The exoskeleton of the tunnel is a series of six degrees of freedom (Six-DOF) articulated links compressible to 1/6 of their fully extended length. The operator can maneuver the control pod to almost any location within about 50 m of the base attachment to the space station. POWER can be envisioned as a series of hollow Six-DOF manipulator segments or arms wherein each arm grasps the shoulder of the next arm. Inside the hollow arms ia a bellow-type access tunnel. The control pod is the fist of the series of linked hollow arms. The fingers of the fist are conventional manipulator arms under direct visual control of the nearby operator in the pod. The applications and progress to date of the POWER system is given.

  6. On the common envelope efficiency

    NASA Astrophysics Data System (ADS)

    Zuo, Zhao-Yu; Li, Xiang-Dong

    2014-08-01

    In this work, we try to use the apparent luminosity versus displacement (i.e. LX versus R) correlation of high-mass X-ray binaries (HMXBs) to constrain the common envelope (CE) efficiency αCE, which is a key parameter affecting the evolution of the binary orbit during the CE phase. The major updates that are crucial for the CE evolution include a variable λ parameter and a new CE criterion for Hertzsprung gap donor stars, both of which are recently developed. We find that, within the framework of the standard energy formula for CE and core definition at mass X = 10 per cent, a high value of αCE, i.e. around 0.8-1.0, is more preferable, while αCE < ˜0.4 likely can not reconstruct the observed LX versus R distribution. However, due to an ambiguous definition for the core boundary in the literature, the used λ here still carries almost two order of magnitude uncertainty, which may translate directly to the expected value of αCE. We present the detailed components of current HMXBs and their spatial offsets from star clusters, which may be further testified by future observations of HMXB populations in nearby star-forming galaxies.

  7. Resource envelope concepts for mission planning

    NASA Technical Reports Server (NTRS)

    Ibrahim, K. Y.; Weiler, J. D.; Tokaz, J. C.

    1991-01-01

    Seven proposed methods for creating resource envelopes for Space Station Freedom mission planning are detailed. Four reference science activity models are used to illustrate the effect of adding operational flexibility to mission timelines. For each method, a brief explanation is given along with graphs to illustrate the application of the envelopes to the power and crew resources. The benefits and costs of each method are analyzed in terms of resource utilization. In addition to the effect on individual activities, resource envelopes are analyzed at the experiment level.

  8. Obstacle-induced spiral vortex breakdown

    NASA Astrophysics Data System (ADS)

    Pasche, Simon; Gallaire, François; Dreyer, Matthieu; Farhat, Mohamed

    2014-08-01

    An experimental investigation on vortex breakdown dynamics is performed. An adverse pressure gradient is created along the axis of a wing-tip vortex by introducing a sphere downstream of an elliptical hydrofoil. The instrumentation involves high-speed visualizations with air bubbles used as tracers and 2D Laser Doppler Velocimeter (LDV). Two key parameters are identified and varied to control the onset of vortex breakdown: the swirl number, defined as the maximum azimuthal velocity divided by the free-stream velocity, and the adverse pressure gradient. They were controlled through the incidence angle of the elliptical hydrofoil, the free-stream velocity and the sphere diameter. A single helical breakdown of the vortex was systematically observed over a wide range of experimental parameters. The helical breakdown coiled around the sphere in the direction opposite to the vortex but rotated along the vortex direction. We have observed that the location of vortex breakdown moved upstream as the swirl number or the sphere diameter was increased. LDV measurements were corrected using a reconstruction procedure taking into account the so-called vortex wandering and the size of the LDV measurement volume. This allows us to investigate the spatio-temporal linear stability properties of the flow and demonstrate that the flow transition from columnar to single helical shape is due to a transition from convective to absolute instability.

  9. Electrical breakdown of water in microgaps

    NASA Astrophysics Data System (ADS)

    Schoenbach, Karl; Kolb, Juergen; Xiao, Shu; Katsuki, Sunao; Minamitani, Yasushi; Joshi, Ravindra

    2008-05-01

    Experimental and modeling studies on electrical breakdown in water in submillimeter gaps between pin and plane electrodes have been performed. Prebreakdown, breakdown and recovery of the water gaps were studied experimentally by using optical and electrical diagnostics with a temporal resolution on the order of one nanosecond. By using Mach-Zehnder interferometry, the electric field distribution in the prebreakdown phase was determined by means of the Kerr effect. Electric fields values in excess of the computed electric fields, which reach >4 MV cm-1 for applied electrical pulses of 20 ns duration, were recorded at the tip of the pin electrode, an effect which can be explained by a reduced permittivity of water at high electric fields. Breakdown of the gaps, streamer-to-arc transition, was recorded by means of high-speed electrical diagnostics, and through high-speed photography. It was shown, through simulations, that breakdown is initiated by field emission at the interface of preexisting microbubbles. Impact ionization within the micro-bubble's gas then contributes to plasma development. Experiments using pulse-probe methods and Schlieren diagnostics allowed us to follow the development of the disturbance caused by the breakdown over a time of more than milliseconds and to determine the recovery time of a water switch. In order to trigger water switches a trigger electrode with a triple point has been utilized. The results of this research have found application in the construction of compact pulse power generators for bioelectric applications.

  10. Intense microwave pulse propagation through gas breakdown plasmas in a waveguide

    SciTech Connect

    Byrne, D.P.

    1986-10-08

    High-power microwave pulse-compression techniques are used to generate 2.856 GHz pulses which are propagated in a TE/sub 10/ mode through a gas filled section of waveguide, where the pulses interact with self-generated gas-breakdown plasmas. Pulse envelopes transmitted through the plasmas, with duration varying from 2 ns to greater than 1 ..mu..s, and peak powers of a few kW to nearly 100 MW, are measured as a function of incident pulse and gas pressure for air, nitrogen, and helium. In addition, the spatial and temporal development of the optical radiation emitted by the breakdown plasmas are measured. For transmitted pulse durations greater than or equal to 100 ns, good agreement is found with both theory and existing measurements. For transmitted pulse duration as short as 2 ns (less than 10 rf cycles), a two-dimensional model is used in which the electrons in the plasma are treated as a fluid whose interactions with the microwave pulse are governed by a self-consistent set of fluid equations and Maxwell's equations for the electromagnetic field. The predictions of this model for air are compared with the experimental results over a pressure range of 0.8 torr to 300 torr. Good agreement is obtained above about 1 torr pressure, demonstrating that microwave pulse propagation above the breakdown threshold can be accurately modeled on this time scale. 63 refs., 44 figs., 2 tabs.

  11. Solar envelope concepts: moderate density building applications

    NASA Astrophysics Data System (ADS)

    Knowles, R. L.; Berry, R. D.

    1980-04-01

    The public policy mechanism for guaranteeing solar access is conceptualized as a solar zoning envelope that allows the largest possible building bulk on a land parcel without shadowing neighboring properties during specified times. Step-by-step methods for generating solar envelopes are described with extensive drawings, showing a variety of urban platting and lot configurations. Development and design possibilities are examined on a selected set of Los Angeles sites with typically diverse urban characteristics. Envelope attributes suitable for encouraging moderate-density commercial and residential building are examined in the context of two hypothetical but realistic development programs: one for speculative office buildings and one for condominium housing. Numerous illustrations of envelope forms and prototypical building designs are provided.

  12. Personnel occupied woven envelope robot power

    NASA Technical Reports Server (NTRS)

    Wessling, F. C.

    1988-01-01

    The Personnel Occupied Woven Envelope Robot (POWER) concept has evolved over the course of the study. The goal of the project was the development of methods and algorithms for solid modeling for the flexible robot arm.

  13. Survival of an Enveloped Virus on Toys.

    PubMed

    Bearden, Richard L; Casanova, Lisa M

    2016-08-01

    Children's toys may carry respiratory viruses. Inactivation of a lipid-enveloped bacteriophage, Φ6, was measured on a nonporous toy at indoor temperature and relative humidity (RH). Inactivation was approximately 2log10 after 24 hours at 60% RH and 6.8log10 at 10 hours at 40% RH. Enveloped viruses can potentially survive on toys long enough to result in exposures. PMID:27144972

  14. Control load envelope shaping by live twist

    NASA Technical Reports Server (NTRS)

    Tarzanin, F. J., Jr.; Mirick, P. H.

    1974-01-01

    Rotor control systems experience a rapid load growth resulting from retreating blade stall during flight conditions of high blade loading or airspeeds. An investigation was undertaken to determine the effect of changing blade torsional properties over the rotor flight envelope. The results of this study show that reducing the blade stiffness to introduce more blade live twist significantly reduces the large retreating blade control loads, while expanding the flight envelope and reducing retreating blade stall loads.

  15. Creating a Lunar EVA Work Envelope

    NASA Technical Reports Server (NTRS)

    Griffin, Brand N.; Howard, Robert; Rajulu, Sudhakar; Smitherman, David

    2009-01-01

    A work envelope has been defined for weightless Extravehicular Activity (EVA) based on the Space Shuttle Extravehicular Mobility Unit (EMU), but there is no equivalent for planetary operations. The weightless work envelope is essential for planning all EVA tasks because it determines the location of removable parts, making sure they are within reach and visibility of the suited crew member. In addition, using the envelope positions the structural hard points for foot restraints that allow placing both hands on the job and provides a load path for reacting forces. EVA operations are always constrained by time. Tasks are carefully planned to ensure the crew has enough breathing oxygen, cooling water, and battery power. Planning first involves computers using a virtual work envelope to model tasks, next suited crew members in a simulated environment refine the tasks. For weightless operations, this process is well developed, but planetary EVA is different and no work envelope has been defined. The primary difference between weightless and planetary work envelopes is gravity. It influences anthropometry, horizontal and vertical mobility, and reaction load paths and introduces effort into doing "overhead" work. Additionally, the use of spacesuits other than the EMU, and their impacts on range of motion, must be taken into account. This paper presents the analysis leading to a concept for a planetary EVA work envelope with emphasis on lunar operations. There is some urgency in creating this concept because NASA has begun building and testing development hardware for the lunar surface, including rovers, habitats and cargo off-loading equipment. Just as with microgravity operations, a lunar EVA work envelope is needed to guide designers in the formative stages of the program with the objective of avoiding difficult and costly rework.

  16. Spectrometers for RF breakdown studies for CLIC

    NASA Astrophysics Data System (ADS)

    Jacewicz, M.; Ziemann, V.; Ekelöf, T.; Dubrovskiy, A.; Ruber, R.

    2016-08-01

    An e+e- collider of several TeV energy will be needed for the precision studies of any new physics discovered at the LHC collider at CERN. One promising candidate is CLIC, a linear collider which is based on a two-beam acceleration scheme that efficiently solves the problem of power distribution to the acceleration structures. The phenomenon that currently prevents achieving high accelerating gradients in high energy accelerators such as the CLIC is the electrical breakdown at very high electrical field. The ongoing experimental work within the CLIC collaboration is trying to benchmark the theoretical models focusing on the physics of vacuum breakdown which is responsible for the discharges. In order to validate the feasibility of accelerating structures and observe the characteristics of the vacuum discharges and their eroding effects on the structure two dedicated spectrometers are now commissioned at the high-power test-stands at CERN. First, the so called Flashbox has opened up a possibility for non-invasive studies of the emitted breakdown currents during two-beam acceleration experiments. It gives a unique possibility to measure the energy of electrons and ions in combination with the arrival time spectra and to put that in context with accelerated beam, which is not possible at any of the other existing test-stands. The second instrument, a spectrometer for detection of the dark and breakdown currents, is operated at one of the 12 GHz stand-alone test-stands at CERN. Built for high repetition rate operation it can measure the spatial and energy distributions of the electrons emitted from the acceleration structure during a single RF pulse. Two new analysis tools: discharge impedance tracking and tomographic image reconstruction, applied to the data from the spectrometer make possible for the first time to obtain the location of the breakdown inside the structure both in the transversal and longitudinal direction thus giving a more complete picture of the

  17. Cooling of neutron stars with diffusive envelopes

    NASA Astrophysics Data System (ADS)

    Beznogov, M. V.; Fortin, M.; Haensel, P.; Yakovlev, D. G.; Zdunik, J. L.

    2016-08-01

    We study the effects of heat blanketing envelopes of neutron stars on their cooling. To this aim, we perform cooling simulations using newly constructed models of the envelopes composed of binary ion mixtures (H-He, He-C, C-Fe) varying the mass of lighter ions (H, He or C) in the envelope. The results are compared with those calculated using the standard models of the envelopes which contain the layers of lighter (accreted) elements (H, He and C) on top of the Fe layer, varying the mass of accreted elements. The main effect is that the chemical composition of the envelopes influences their thermal conductivity and, hence, thermal insulation of the star. For illustration, we apply these results to estimate the internal temperature of the Vela pulsar and to study the cooling of neutron stars of ages of 105 - 106 yr at the photon cooling stage. The uncertainties of the cooling models associated with our poor knowledge of chemical composition of the heat insulating envelopes strongly complicate theoretical reconstruction of the internal structure of cooling neutron stars from observations of their thermal surface emission.

  18. Genetic diversity of koala retroviral envelopes.

    PubMed

    Xu, Wenqin; Gorman, Kristen; Santiago, Jan Clement; Kluska, Kristen; Eiden, Maribeth V

    2015-03-01

    Genetic diversity, attributable to the low fidelity of reverse transcription, recombination and mutation, is an important feature of infectious retroviruses. Under selective pressure, such as that imposed by superinfection interference, gammaretroviruses commonly adapt their envelope proteins to use alternative receptors to overcome this entry block. The first characterized koala retroviruses KoRV subgroup A (KoRV-A) were remarkable in their absence of envelope genetic variability. Once it was determined that KoRV-A was present in all koalas in US zoos, regardless of their disease status, we sought to isolate a KoRV variant whose presence correlated with neoplastic malignancies. More than a decade after the identification of KoRV-A, we isolated a second subgroup of KoRV, KoRV-B from koalas with lymphomas. The envelope proteins of KoRV-A and KoRV-B are sufficiently divergent to confer the ability to bind and employ distinct receptors for infection. We have now obtained a number of additional KoRV envelope variants. In the present studies we report these variants, and show that they differ from KoRV-A and KoRV-B envelopes in their host range and superinfection interference properties. Thus, there appears to be considerable variation among KoRVs envelope genes suggesting genetic diversity is a factor following the KoRV-A infection process.

  19. Genetic diversity of koala retroviral envelopes.

    PubMed

    Xu, Wenqin; Gorman, Kristen; Santiago, Jan Clement; Kluska, Kristen; Eiden, Maribeth V

    2015-03-01

    Genetic diversity, attributable to the low fidelity of reverse transcription, recombination and mutation, is an important feature of infectious retroviruses. Under selective pressure, such as that imposed by superinfection interference, gammaretroviruses commonly adapt their envelope proteins to use alternative receptors to overcome this entry block. The first characterized koala retroviruses KoRV subgroup A (KoRV-A) were remarkable in their absence of envelope genetic variability. Once it was determined that KoRV-A was present in all koalas in US zoos, regardless of their disease status, we sought to isolate a KoRV variant whose presence correlated with neoplastic malignancies. More than a decade after the identification of KoRV-A, we isolated a second subgroup of KoRV, KoRV-B from koalas with lymphomas. The envelope proteins of KoRV-A and KoRV-B are sufficiently divergent to confer the ability to bind and employ distinct receptors for infection. We have now obtained a number of additional KoRV envelope variants. In the present studies we report these variants, and show that they differ from KoRV-A and KoRV-B envelopes in their host range and superinfection interference properties. Thus, there appears to be considerable variation among KoRVs envelope genes suggesting genetic diversity is a factor following the KoRV-A infection process. PMID:25789509

  20. Breakdowns in Coordination Between Air Traffic Controllers

    NASA Technical Reports Server (NTRS)

    Bearman, Chris; Orasanu, Judith; Miller, Ronald C.

    2011-01-01

    This talk outlines the complexity of coordination in air traffic control, introduces the NextGen technologies, identifies common causes for coordination breakdowns in air traffic control and examines whether these causes are likely to be reduced with the introduction of NextGen technologies. While some of the common causes of breakdowns will be reduced in a NextGen environment this conclusion should be drawn carefully given the current stage of development of the technologies and the observation that new technologies often shift problems rather than reduce them.

  1. Nuclear removal during terminal lens fiber cell differentiation requires CDK1 activity: appropriating mitosis-related nuclear disassembly

    PubMed Central

    Chaffee, Blake R.; Shang, Fu; Chang, Min-Lee; Clement, Tracy M.; Eddy, Edward M.; Wagner, Brad D.; Nakahara, Masaki; Nagata, Shigekazu; Robinson, Michael L.; Taylor, Allen

    2014-01-01

    Lens epithelial cells and early lens fiber cells contain the typical complement of intracellular organelles. However, as lens fiber cells mature they must destroy their organelles, including nuclei, in a process that has remained enigmatic for over a century, but which is crucial for the formation of the organelle-free zone in the center of the lens that assures clarity and function to transmit light. Nuclear degradation in lens fiber cells requires the nuclease DNase IIβ (DLAD) but the mechanism by which DLAD gains access to nuclear DNA remains unknown. In eukaryotic cells, cyclin-dependent kinase 1 (CDK1), in combination with either activator cyclins A or B, stimulates mitotic entry, in part, by phosphorylating the nuclear lamin proteins leading to the disassembly of the nuclear lamina and subsequent nuclear envelope breakdown. Although most post-mitotic cells lack CDK1 and cyclins, lens fiber cells maintain these proteins. Here, we show that loss of CDK1 from the lens inhibited the phosphorylation of nuclear lamins A and C, prevented the entry of DLAD into the nucleus, and resulted in abnormal retention of nuclei. In the presence of CDK1, a single focus of the phosphonuclear mitotic apparatus is observed, but it is not focused in CDK1-deficient lenses. CDK1 deficiency inhibited mitosis, but did not prevent DNA replication, resulting in an overall reduction of lens epithelial cells, with the remaining cells possessing an abnormally large nucleus. These observations suggest that CDK1-dependent phosphorylations required for the initiation of nuclear membrane disassembly during mitosis are adapted for removal of nuclei during fiber cell differentiation. PMID:25139855

  2. Comment on the invariant envelope solution in rf photoinjectors.

    SciTech Connect

    Wang, C.-x.; Accelerator Systems Division

    2006-02-01

    The beam envelope equation has been used to address the beam dynamics in rf photoinjectors. A special solution of the envelope equation, known as the invariant envelope, plays a critical role in the theory of emittance compensation. In this comment, I will present a different view of the invariant envelope solution that better delineates its properties and simplifies the picture of beam dynamics.

  3. Simulating Convection in Stellar Envelopes

    NASA Astrophysics Data System (ADS)

    Tanner, Joel

    Understanding convection in stellar envelopes, and providing a mathematical description of it, would represent a substantial advance in stellar astrophysics. As one of the largest sources of uncertainty in stellar models, existing treatments of convection fail to account for many of the dynamical effects of convection, such as turbulent pressure and asymmetry in the velocity field. To better understand stellar convection, we must be able to study and examine it in detail, and one of the best tools for doing so is numerical simulation. Near the stellar surface, both convective and radiative process play a critical role in determining the structure and gas dynamics. By following these processes from first principles, convection can be simulated self-consistently and accurately, even in regions of inefficient energy transport where existing descriptions of convection fail. Our simulation code includes two radiative transfer solvers that are based on different assumptions and approximations. By comparing simulations that differ only in their respective radiative transfer methods, we are able to isolate the effect that radiative efficiency has on the structure of the superadiabatic layer. We find the simulations to be in good general agreement, but they show distinct differences in the thermal structure in the superadiabatic layer and atmosphere. Using the code to construct a grid of three-dimensional radiation hydrodynamic simulations, we investigate the link between convection and various chemical compositions. The stellar parameters correspond to main-sequence stars at several surface gravities, and span a range in effective temperatures (4500 < Teff < 6400). Different chemical compositions include four metallicities (Z = 0.040, 0.020, 0.010, 0.001), three helium abundances (Y = 0.1, 0.2, 0.3) and several levels of alpha-element enhancement. Our grid of simulations shows that various convective properties, such as velocity and the degree of superadiabaticity, are

  4. RF Breakdown of Metallic Surfaces in Hydrogen

    SciTech Connect

    BastaniNejad, M.; Elmustafa, A.A.; Yonehara, K.; Chung, M.; Jansson, A.; Hu, M.; Moretti, A.; Popovic, M.; Alsharo'a, M.; Neubauer, M.; Sah, R.; /Muons Inc., Batavia

    2009-05-01

    In earlier reports, microscopic images of the surfaces of metallic electrodes used in high-pressure gas-filled 805 MHz RF cavity experiments were used to investigate the mechanism of RF breakdown of tungsten, molybdenum, and beryllium electrode surfaces. Plots of remnants were consistent with the breakdown events being due to field emission, due to the quantum mechanical tunnelling of electrons through a barrier as described by Fowler and Nordheim. In the work described here, these studies have been extended to include tin, aluminium, and copper. Contamination of the surfaces, discovered after the experiments concluded, have cast some doubt on the proper qualities to assign to the metallic surfaces. However, two significant results are noted. First, the maximum stable RF gradient of contaminated copper electrodes is higher than for a clean surface. Second, the addition of as little as 0.01% of SF6 to the hydrogen gas increased the maximum stable gradient, which implies that models of RF breakdown in hydrogen gas will be important to the study of metallic breakdown.

  5. Fear of breakdown and the unlived life.

    PubMed

    Ogden, Thomas H

    2014-04-01

    Winnicott's Fear of breakdown is an unfinished work that requires that the reader be not only a reader, but also a writer of this work which often gestures toward meaning as opposed to presenting fully developed ideas. The author's understanding of the often confusing, sometimes opaque, argument of Winnicott's paper is as follows. In infancy there occurs a breakdown in the mother-infant tie that forces the infant to take on, by himself, emotional events that he is unable to manage. He short-circuits his experience of primitive agony by generating defense organizations that are psychotic in nature, i.e., they substitute self-created inner reality for external reality, thus foreclosing his actually experiencing critical life events. By not experiencing the breakdown of the mother-infant tie when it occurred in infancy, the individual creates a psychological state in which he lives in fear of a breakdown that has already happened, but which he did not experience. The author extends Winnicott's thinking by suggesting that the driving force of the patient's need to find the source of his fear is his feeling that parts of himself are missing and that he must find them if he is to become whole. What remains of his life feels to him like a life that is mostly an unlived life. PMID:24620827

  6. Heme content and breakdown in developing chloroplasts

    SciTech Connect

    Thomas, J.; Weinstein, J.D. )

    1990-05-01

    Heme regulates tetrapyrrole biosynthesis in plants by inhibition of {delta}-aminolevulinic acid (ALA) synthesis, product inhibition of heme synthesis, and possibly other mechanisms. Plastid heme levels may be modulated by heme synthesis, breakdown and/or efflux. Heme breakdown may be catalyzed by a chloroplast localized heme oxygenase. Chloroplasts isolated from greening cucumber cotyledons were incubated in the presence or absence of various components thought to modulate heme breakdown. Following the incubations, the chloroplasts were broken (freeze-thaw) and then supplemented with horseradish peroxidase apoenzyme. The reconstituted peroxidase activity was used to determine the amount of free heme remaining (Thomas Weinstein (1989) Plant Physiol. 89S: 74). Chloroplasts, freshly isolated from seedlings greened for 16 hours, contained approximately 37 pmol heme/mg protein. When chloroplasts were incubated with 5 mM NADPH for 30 min, the endogenous heme dropped to unmeasurable levels. Exogenous heme was also broken down when NADPH was included in the incubation. Heme levels could be increased by the inclusion of 50 {mu}M ALA and/or p-hydroxymercuribenzoate. The increase due to exogenous ALA was blocked by levulinic acid, an inhibitor of ALA utilization. NADPH-dependent heme breakdown acid was inhibited by p-hydroxymercuribenzoate.

  7. Numerical Borehole Breakdown Investigations using XFEM

    NASA Astrophysics Data System (ADS)

    Beckhuis, Sven; Leonhart, Dirk; Meschke, Günther

    2016-04-01

    During pressurization of a wellbore a typical downhole pressure record shows the following regimes: first the applied wellbore pressure balances the reservoir pressure, then after the compressive circumferential hole stresses are overcome, tensile stresses are induced on the inside surface of the hole. When the magnitude of these stresses reach the tensile failure stress of the surrounding rock medium, a fracture is initiated and propagates into the reservoir. [1] In standard theories this pressure, the so called breakdown pressure, is the peak pressure in the down-hole pressure record. However experimental investigations [2] show that the breakdown did not occur even if a fracture was initiated at the borehole wall. Drilling muds had the tendency to seal and stabilize fractures and prevent fracture propagation. Also fracture mechanics analysis of breakdown process in mini-frac or leak off tests [3] show that the breakdown pressure could be either equal or larger than the fracture initiation pressure. In order to gain a deeper understanding of the breakdown process in reservoir rock, numerical investigations using the extended finite element method (XFEM) for hydraulic fracturing of porous materials [4] are discussed. The reservoir rock is assumed to be pre-fractured. During pressurization of the borehole, the injection pressure, the pressure distribution and the position of the highest flux along the fracture for different fracturing fluid viscosities are recorded and the influence of the aforementioned values on the stability of fracture propagation is discussed. [1] YEW, C. H. (1997), "Mechanics of Hydraulic Fracturing", Gulf Publishing Company [2] MORITA, N.; BLACK, A. D.; FUH, G.-F. (1996), "Borehole Breakdown Pressure with Drilling Fluids". International Journal of Rock Mechanics and Mining Sciences 33, pp. 39-51 [3] DETOURNAY, E.; CARBONELL, R. (1996), "Fracture Mechanics Analysis of the Breakdown Process in Minifrac or Leakoff Test", Society of Petroleum

  8. Safeguards Envelope: The First Steps

    SciTech Connect

    Richard Metcalf; Jean Ragusa; Robert Bean

    2008-03-01

    The possibility exists for real time accountancy and assay of nuclear materials as they move through a reprocessing facility. This project aims to establish working parameters and local figures of merit to identify possible diversion in real time with minimal operational impact. Factors such as pH, NOX gas concentration, flow speeds and radiation fields are rarely taken into account in safeguards methodologies and will be included to increase the confidence of location and assay of nuclear materials. An adaptable, real data model is being created of the contactors of the Advanced Fuel Cycle Facility and will be analyzed using the appropriate modeling codes. This model will then be subjected to three, diversion scenarios and a figure of merit methodology will be utilized to create the operational parameters under which these diversion scenarios would be detected. This analysis for figure of merit methodology will include statistical fluctuations, operator error, and a rudimentary analysis of transient conditions. The long term goal of the project includes expansion universally over the plant, methods of detection without requiring access to proprietary information, and an evaluation of the requirements for future figure of merit methodologies.

  9. The Latitude Dependence of Dielectric Breakdown on the Moon

    NASA Astrophysics Data System (ADS)

    Jordan, A. P.; Stubbs, T. J.; Wilson, J. K.; Hayne, P. O.; Schwadron, N. A.; Spence, H. E.; Izenberg, N. R.

    2016-11-01

    Solar energetic particles may cause dielectric breakdown on the nightside of the Moon. We predict that breakdown weathering may have melted or vaporized about 4-11 wt% of impact gardened regolith on the Moon.

  10. Electrical Breakdown of Plasma-Polymerized Styrene Thin Films

    NASA Astrophysics Data System (ADS)

    Hikita, Masayuki; Matsuda, Akinori; Nagao, Masayuki; Sawa, Goro; Ieda, Masayuki

    1982-03-01

    The electrical breakdown of plasma-polymerized styrene thin film (PPS) was studied by taking advantage of self-healing. The electric strength FB was almost independent of temperature from -196 to 200°C, and strongly depended on the rate of voltage increase even at a slow rate of increase. The breakdown characteristics were influenced by the electrode metal and the ambient atmosphere, but not by X-ray irradiation or photoillumination. The experimental results are used to discuss the breakdown mechanism of PPS through existing breakdown theories. As a result, no single breakdown process was considered as a possible breakdown mechanism, and we thus obtained important conditions for presenting a new breakdown model; the breakdown of PPS will be determined by a thermal criterion, and it will be closely related to a temperature-independent injection process.

  11. Morphologically complex protostellar envelopes : structure and kinematics

    NASA Astrophysics Data System (ADS)

    Tobin, John J.

    I present an in-depth study of protostars and their surrounding envelopes of dense gas and dust, using a multitude of observational methods to reveal new details of the star formation process. I use mid-infrared imaging from the Spitzer Space Telescope, combined with photometry spanning the near-infrared to millimeter wavelengths, to construct a model of the L1527 protostellar system. I modeled both the spectral energy distribution and resolved scattered light images to determine physical properties of the protostellar system. The nature of the apparent central point source in the Spitzer images was uncertain until high-resolution L-band imaging from the Gemini observatory resolved the point source into a disk in scattered light, having a radius of 200 AU. Protostellar envelopes are also often found to cast shadows against the 8 micron Galactic background in Spitzer imaging, enabling direct probes of envelope structure. The shadow images show that the dense envelopes around twenty-two Class 0 protostars are generally morphologically complex from 0.1 pc scales down to ˜1000 AU; they are often filamentary, and frequently non-axisymmetric. The observed envelope structure indicates a likely origin in turbulent cloud structure rather than a quasi-static/equilibrium formation. The complex envelope structure also may indicate an increased likelihood of fragmentation during collapse, forming close binaries. To further characterize these envelopes, I have observed them in the dense molecular gas tracers nthp and nht, both of which closely follow the 8 micron extinction morphology. The magnitude of the velocity gradients and envelope complexity on ˜10000 AU scales indicates that the velocity structure may reflect large-scale infall in addition to the often assumed rotation. Comparisons with three-dimensional filamentary and symmetric rotating collapse models reinforce the interpretation of velocities reflecting large-scale infall, showing that the structure of the envelope

  12. A computational study of the taxonomy of vortex breakdown

    NASA Technical Reports Server (NTRS)

    Spall, Robert E.; Gatski, Thomas B.

    1990-01-01

    The results of a fully three-dimensional numerical simulation of vortex breakdown using the unsteady, incompressible Navier-Stokes equations are presented. The solutions show that the freestream axial velocity distribution has a significant effect on the position and type of vortex breakdown. Common features between bubble-type and spiral-type breakdown are identified and the role of flow stagnation and the critical state are discussed as complimentary ideas describing the initiation of breakdown.

  13. Featured Image: Orbiting Stars Share an Envelope

    NASA Astrophysics Data System (ADS)

    Kohler, Susanna

    2016-03-01

    This beautiful series of snapshots from a simulation (click for a better look!) shows what happens when two stars in a binary system become enclosed in the same stellar envelope. In this binary system, one of the stars has exhausted its hydrogen fuel and become a red giant, complete with an expanding stellar envelope composed of hydrogen and helium. Eventually, the envelope expands so much that the companion star falls into it, where it releases gravitational potential energy into the common envelope. A team led by Sebastian Ohlmann (Heidelberg Institute for Theoretical Studies and University of Wrzburg) recently performed hydrodynamic simulations of this process. Ohlmann and collaborators discovered that the energy release eventually triggers large-scale flow instabilities, which leads to turbulence within the envelope. This process has important consequences for how these systems next evolve (for instance, determining whether or not a supernova occurs!). You can check out the authors video of their simulated stellar inspiral below, or see their paper for more images and results from their study.CitationSebastian T. Ohlmann et al 2016 ApJ 816 L9. doi:10.3847/2041-8205/816/1/L9

  14. The cell envelope proteome of Aggregatibacter actinomycetemcomitans

    PubMed Central

    Smith, Kenneth P.; Fields, Julia G.; Voogt, Richard D.; Deng, Bin; Lam, Ying-Wai; Mintz, Keith P.

    2014-01-01

    Summary The cell envelope of Gram-negative bacteria serves a critical role in maintenance of cellular homeostasis, resistance to external stress, and host-pathogen interactions. Envelope protein composition is influenced by the physiological and environmental demands placed on the bacterium. In this study, we report a comprehensive compilation of cell envelope proteins from the periodontal and systemic pathogen Aggregatibacter actinomycetemcomitans VT1169, an afimbriated serotype b strain. The urea-extracted membrane proteins were identified by mass spectrometry-based shotgun proteomics. The membrane proteome, isolated from actively growing bacteria under normal laboratory conditions, included 648 proteins representing 28% of the predicted ORFs in the genome. Bioinformatic analyses were used to annotate and predict the cellular location and function of the proteins. Surface adhesins, porins, lipoproteins, numerous influx and efflux pumps, multiple sugar, amino acid and iron transporters, and components of the type I, II and V secretion systems were identified. Periplasmic space and cytoplasmic proteins with chaperone function were also identified. 107 proteins with unknown function were associated with the cell envelope. Orthologs of a subset of these uncharacterized proteins are present in other bacterial genomes, while others are found exclusively in A. actinomycetemcomitans. This knowledge will contribute to elucidating the role of cell envelope proteins in bacterial growth and survival in the oral cavity. PMID:25055881

  15. Envelope Solitons in Acoustically Dispersive Vitreous Silica

    NASA Technical Reports Server (NTRS)

    Cantrell, John H.; Yost, William T.

    2012-01-01

    Acoustic radiation-induced static strains, displacements, and stresses are manifested as rectified or dc waveforms linked to the energy density of an acoustic wave or vibrational mode via the mode nonlinearity parameter of the material. An analytical model is developed for acoustically dispersive media that predicts the evolution of the energy density of an initial waveform into a series of energy solitons that generates a corresponding series of radiation-induced static strains (envelope solitons). The evolutionary characteristics of the envelope solitons are confirmed experimentally in Suprasil W1 vitreous silica. The value (-11.9 plus or minus 1.43) for the nonlinearity parameter, determined from displacement measurements of the envelope solitons via a capacitive transducer, is in good agreement with the value (-11.6 plus or minus 1.16) obtained independently from acoustic harmonic generation measurements. The agreement provides strong, quantitative evidence for the validity of the model.

  16. Prevention of breakdown behind railgun projectiles

    DOEpatents

    Hawke, Ronald S.

    1992-01-01

    An electromagnetic railgun accelerator system, for accelerating projectiles (14, 15, 114, 214, 314, 444) by a plasma arc (3), introduces a breakdown inhibiting gas into the railgun chamber (26) behind the accelerating projectile (14). The breakdown inhibiting gas, which absorbs electrons, is a halide or a halide compound such as fluorine or SF.sub.6. The gas is introduced between the railgun rails (12) after the projectile (14) has passed through inlets (16) in the rails (12) or the projectile (114); by coating the rails (12) or the projectile (15) with a material (28) which releases the gas after the projectile (14 ) passes over it; by fabricating the rails (12) or the projectile (15) or insulators out of a material which releases the gas into the portions of the chamber (26) through which the projectile has travelled. The projectile (214, 314, 414) may have a cavity (232, 332, 432) at its rear to control the release of ablation products (4).

  17. Prevention of breakdown behind railgun projectiles

    DOEpatents

    Hawke, Ronald S.

    1992-01-01

    An electromagnetic railgun accelerator system, for accelerating projectiles (14, 15, 114, 214, 314, 414) by a plasma arc (3), introduces a breakdown inhibiting gas into the railgun chamber (26) behind the accelerating projectile (14). The breakdown inhibiting gas, which absorbs electrons, is a halide or a halide compound such as fluorine or SF.sub.6. The gas is introduced between the railgun rails (12) after the projectile (14) has passed through inlets (16) in the rails (12) or the projectile (114); by coating the rails (12) or the projectile (15) with a material (28) which releases the gas after the projectile (14) passes over it; by fabricating the rails (12) or the projectile (15) or insulators out of a material which releases the gas into the portions of the chamber (26) through which the projectile has travelled. The projectile (214, 314, 414) may have a cavity (232, 332, 432) at its rear to control the release of ablation products (4).

  18. Prevention of breakdown behind railgun projectiles

    DOEpatents

    Hawke, R.S.

    1992-09-01

    An electromagnetic railgun accelerator system, for accelerating projectiles by a plasma arc, introduces a breakdown inhibiting gas into the railgun chamber behind the accelerating projectile. The breakdown inhibiting gas, which absorbs electrons, is a halide or a halide compound such as fluorine or SF[sub 6]. The gas is introduced between the railgun rails after the projectile has passed through inlets in the rails or the projectile; by coating the rails or the projectile with a material which releases the gas after the projectile passes over it; by fabricating the rails or the projectile or insulators out of a material which releases the gas into the portions of the chamber through which the projectile has travelled. The projectile may have a cavity at its rear to control the release of ablation products. 12 figs.

  19. Prevention of breakdown behind railgun projectiles

    DOEpatents

    Hawke, R.S.

    1992-10-13

    An electromagnetic railgun accelerator system, for accelerating projectiles by a plasma arc, introduces a breakdown inhibiting gas into the railgun chamber behind the accelerating projectile. The breakdown inhibiting gas, which absorbs electrons, is a halide or a halide compound such as fluorine or SF[sub 6]. The gas is introduced between the railgun rails after the projectile has passed through inlets in the rails or the projectile; by coating the rails or the projectile with a material which releases the gas after the projectile passes over it; by fabricating the rails or the projectile or insulators out of a material which releases the gas into the portions of the chamber through which the projectile has travelled. The projectile may have a cavity at its rear to control the release of ablation products. 12 figs.

  20. Electrical breakdown studies with Mycalex insulators

    SciTech Connect

    Waldron, W.; Greenway, W.; Eylon, S.; Henestroza, E.; Yu, S.

    2003-05-01

    Insulating materials such as alumina and glass-bonded mica (Mycalex) are used in accelerator systems for high voltage feedthroughs, structural supports, and barriers between high voltage insulating oil and the vacuum beam pipe in induction accelerator cells. Electric fields in the triple points should be minimized to prevent voltage breakdown. Mechanical stress can compromise seals and result in oil contamination of the insulator surface. We have tested various insulator cleaning procedures including ultrasonic cleaning with a variety of aqueous-based detergents, and manual scrubbing with various detergents. Water sheeting tests were used to determine the initial results of the cleaning methods. Ultimately, voltage breakdown tests will be used to quantify the benefits of these cleaning procedures.

  1. Breakdown and partial discharges in magnetic liquids.

    PubMed

    Herchl, F; Marton, K; Tomčo, L; Kopčanský, P; Timko, M; Koneracká, M; Kolcunová, I

    2008-05-21

    The dielectric properties (permittivity, loss factor, dielectric breakdown strength) of magnetic liquids were investigated. The magnetic liquids were composed of magnetite particles coated with oleic acid as surfactant and dispersed in transformer oil. To determine their dielectric properties they were subjected to a uniform magnetic field at high alternating electric fields up to 14 MV m(-1). Nearly constant permittivity of magnetic liquid with particle volume concentration Φ = 0.0019 as a function of electric field was observed. Magnetic liquids with concentrations Φ = 0.019 and 0.032 showed significant changes of permittivity and loss factor dependent on electric and magnetic fields. The best concentration of magnetic fluid was found at which partial current impulse magnitudes were the lowest. The breakdown strength distribution of the magnetic liquid with Φ = 0.0025 was fitted with the Duxbury-Leath, Weibull and Gauss distribution functions. PMID:21694240

  2. Dynamic Remodeling of the Plastid Envelope Membranes – A Tool for Chloroplast Envelope in vivo Localizations

    PubMed Central

    Breuers, Frederique K. H.; Bräutigam, Andrea; Geimer, Stefan; Welzel, Ulla Y.; Stefano, Giovanni; Renna, Luciana; Brandizzi, Federica; Weber, Andreas P. M.

    2012-01-01

    Two envelope membranes delimit plastids, the defining organelles of plant cells. The inner and outer envelope membranes are unique in their protein and lipid composition. Several studies have attempted to establish the proteome of these two membranes; however, differentiating between them is difficult due to their close proximity. Here, we describe a novel approach to distinguish the localization of proteins between the two membranes using a straightforward approach based on live cell imaging coupled with transient expression. We base our approach on analyses of the distribution of GFP-fusions, which were aimed to verify outer envelope membrane proteomics data. To distinguish between outer envelope and inner envelope protein localization, we used AtTOC64–GFP and AtTIC40–GFP, as respective controls. During our analyses, we observed membrane proliferations and loss of chloroplast shape in conditions of protein over-expression. The morphology of the proliferations varied in correlation with the suborganellar distribution of the over-expressed proteins. In particular, while layers of membranes built up in the inner envelope membrane, the outer envelope formed long extensions into the cytosol. Using electron microscopy, we showed that these extensions were stromules, a dynamic feature of plastids. Since the behavior of the membranes is different and is related to the protein localization, we propose that in vivo studies based on the analysis of morphological differences of the membranes can be used to distinguish between inner and outer envelope localizations of proteins. To demonstrate the applicability of this approach, we demonstrated the localization of AtLACS9 to the outer envelope membrane. We also discuss protein impact on membrane behavior and regulation of protein insertion into membranes, and provide new hypotheses on the formation of stromules. PMID:22645566

  3. Live Cell Dynamics of Promyelocytic Leukemia Nuclear Bodies upon Entry into and Exit from Mitosis

    PubMed Central

    Chen, Yi-Chun M.; Kappel, Constantin; Beaudouin, Joel; Eils, Roland

    2008-01-01

    Promyelocytic leukemia nuclear bodies (PML NBs) have been proposed to be involved in tumor suppression, viral defense, DNA repair, and/or transcriptional regulation. To study the dynamics of PML NBs during mitosis, we developed several U2OS cell lines stably coexpressing PML-enhanced cyan fluorescent protein with other individual marker proteins. Using three-dimensional time-lapse live cell imaging and four-dimensional particle tracking, we quantitatively demonstrated that PML NBs exhibit a high percentage of directed movement when cells progressed from prophase to prometaphase. The timing of this increased dynamic movement occurred just before or upon nuclear entry of cyclin B1, but before nuclear envelope breakdown. Our data suggest that entry into prophase leads to a loss of tethering between regions of chromatin and PML NBs, resulting in their increased dynamics. On exit from mitosis, Sp100 and Fas death domain-associated protein (Daxx) entered the daughter nuclei after a functional nuclear membrane was reformed. However, the recruitment of these proteins to PML NBs was delayed and correlated with the timing of de novo PML NB formation. Together, these results provide insight into the dynamic changes associated with PML NBs during mitosis. PMID:18480407

  4. Interchangeability of vortex-breakdown types

    NASA Astrophysics Data System (ADS)

    Kurosaka, M.; Kikuchi, M.; Hirano, K.; Yuge, T.; Inoue, H.

    In order to investigate the connection between the bubble and the spiral form of vortex breakdown, experiments were conducted: an external disturbance in the form of an azimuthally spinning waveform was imposed in a pipe. The azimuthal wave number was varied by adjusting the phase difference among four oscillating pistons mounted circumferentially on the pipe. By imposing a disturbance of zero azimuthal wave number, a spiral was transformed into a bubble, and this occurred only for selective piston frequencies; the vortex breakdown which altered from the spiral to the bubble moved upstream, where it remained as a bubble as long as the external disturbance remained. Once the disturbance was removed, the bubble returned to a spiral. By imposing a disturbance of azimuthal wave number +1 (the first circumferential mode rotating in the same direction as the mean swirl), a bubble was transformed into a spiral for selective piston frequencies, and the spiral moved downstream. These preferred frequencies were found to be the same as the unexcited frequencies observed in the spiral in its natural state. As long as the external disturbance was imposed, the breakdown altered from the bubble to the spiral remained as a spiral; once the disturbance was removed, the spiral reverted to a bubble. By imposing a disturbance with azimuthal wave number -1 (the first circumferential mode rotating in the opposite direction to the mean swirl), no change was detected in either a bubble or a spiral. By imposing a disturbance with azimuthal wave number 2 (the second circumferential mode), for selective piston frequencies a bubble was transformed into what appears to be the so-called two-tailed type. Thus, it appears that hydrodynamic instability plays a role in interchanging vortex breakdown types, and a comparison with available stability theories is discussed.

  5. Locating Initial Breakdown Pulses of Lightning Flashes

    NASA Astrophysics Data System (ADS)

    Karunarathne, S.; Marshall, T.; Stolzenburg, M.; Betz, H.; Wieczorek, G.

    2010-12-01

    Lightning flashes often begin with a series of bipolar pulses, 1-5 us in width, called initial breakdown pulses or characteristic pulses. In this presentation we show electric field change data of initial breakdown pulses collected with a network of 5 flat-plate antennas with a bandwidth of 0 - 5 MHz. These pulses were obtained at the NASA/Kennedy Space Center (KSC) during the summer of 2010. The (x, y, z, t) positions of these pulses have been determined using a time of arrival technique [Koshak and Solakiewicz, JGR, 1996] for several lightning flashes. In addition, we also collected magnetic field change data with a LINET system [e.g., Betz et al., GRL, 2004], which consisted of 7 crossed-loop sensors having a bandwidth of 5 - 200 kHz; the pulse locations detected by this system were also determined by time of arrival. The locations of the initial breakdown pulses from both systems will be compared to locations of VHF lightning sources made with the KSC LDAR2 system (with a center frequency of 63 MHz and a bandwidth of 6 MHz). Possible implications of the pulse locations derived from the three different sets of sensors on lightning initiation and propagation will be discussed.

  6. Kinetic Simulations of Dense Plasma Focus Breakdown

    NASA Astrophysics Data System (ADS)

    Schmidt, A.; Higginson, D. P.; Jiang, S.; Link, A.; Povilus, A.; Sears, J.; Bennett, N.; Rose, D. V.; Welch, D. R.

    2015-11-01

    A dense plasma focus (DPF) device is a type of plasma gun that drives current through a set of coaxial electrodes to assemble gas inside the device and then implode that gas on axis to form a Z-pinch. This implosion drives hydrodynamic and kinetic instabilities that generate strong electric fields, which produces a short intense pulse of x-rays, high-energy (>100 keV) electrons and ions, and (in deuterium gas) neutrons. A strong factor in pinch performance is the initial breakdown and ionization of the gas along the insulator surface separating the two electrodes. The smoothness and isotropy of this ionized sheath are imprinted on the current sheath that travels along the electrodes, thus making it an important portion of the DPF to both understand and optimize. Here we use kinetic simulations in the Particle-in-cell code LSP to model the breakdown. Simulations are initiated with neutral gas and the breakdown modeled self-consistently as driven by a charged capacitor system. We also investigate novel geometries for the insulator and electrodes to attempt to control the electric field profile. The initial ionization fraction of gas is explored computationally to gauge possible advantages of pre-ionization which could be created experimentally via lasers or a glow-discharge. Prepared by LLNL under Contract DE-AC52-07NA27344.

  7. Runaway breakdown and electrical discharges in thunderstorms

    NASA Astrophysics Data System (ADS)

    Milikh, Gennady; Roussel-Dupré, Robert

    2010-12-01

    This review considers the precise role played by runaway breakdown (RB) in the initiation and development of lightning discharges. RB remains a fundamental research topic under intense investigation. The question of how lightning is initiated and subsequently evolves in the thunderstorm environment rests in part on a fundamental understanding of RB and cosmic rays and the potential coupling to thermal runaway (as a seed to RB) and conventional breakdown (as a source of thermal runaways). In this paper, we describe the basic mechanism of RB and the conditions required to initiate an observable avalanche. Feedback processes that fundamentally enhance RB are discussed, as are both conventional breakdown and thermal runaway. Observations that provide clear evidence for the presence of energetic particles in thunderstorms/lightning include γ-ray and X-ray flux intensifications over thunderstorms, γ-ray and X-ray bursts in conjunction with stepped leaders, terrestrial γ-ray flashes, and neutron production by lightning. Intense radio impulses termed narrow bipolar pulses (or NBPs) provide indirect evidence for RB particularly when measured in association with cosmic ray showers. Our present understanding of these phenomena and their enduring enigmatic character are touched upon briefly.

  8. Breakdown analysis of multilayer amorphous silicon photoreceptors

    NASA Astrophysics Data System (ADS)

    Hu, Jian

    1993-06-01

    The breakdown mechanism of hydrogenated amorphous silicon (a-Si:H) has been investigated. It has been shown that the acceptance of the surface potential of an a-Si:H photoreceptor is very sensitive to the micro-roughness of the substrate surface. This is because the junction between the metal substrate (usually aluminum) and the blocking layer (p+ or n+ a-Si:H) is strongly affected by the micro-roughness of the substrate surface. A model is proposed to expound this phenomenon, which indicates that the existence of micro- defects on the substrate surface results in the bending of the metal-semiconductor junction at these defect positions; that is, the original parallel plane junction changes into a spherical abrupt junction. Compared to the former, the curved junction has a lower breakdown voltage, therefore, it will more easily break down at these defect positions during charging. An a-Si:H photoreceptor was prepared on the drum substrate half covered with a thin aluminum film to confirm the model. The experiment result was qualitatively in agreement with the analysis mentioned above. In addition, the effects of PVD-like deposition processes (e.g., high power or high argon diluted silane deposition) on the microstructure and breakdown of a-Si:H photoreceptors are reviewed.

  9. Dielectric breakdown induced by picosecond laser pulses

    NASA Technical Reports Server (NTRS)

    Smith, W. L.; Bechtel, J. H.; Bloembergen, N.

    1976-01-01

    The damage thresholds of transparent optical materials were investigated. Single picosecond pulses at 1.06 microns, 0.53 microns and 0.35 microns were obtained from a mode locked Nd-YAG oscillator-amplifier-frequency multiplier system. The pulses were Gaussian in space and time and permitted the determination of breakdown thresholds with a reproducibility of 15%. It was shown that the breakdown thresholds are characteristic of the bulk material, which included nine alkali halides, five different laser host materials, KDP, quartz, sapphire and calcium fluoride. The extension of the damage data to the ultraviolet is significant, because some indication was obtained that two- and three-photon absorption processes begin to play a role in determining the threshold. Throughout the visible region of the spectrum the threshold is still an increasing function of frequency, indicating that avalanche ionization is the dominant factor in determining the breakdown threshold. This was confirmed by a detailed study of the damage morphology with a high resolution microscope just above the threshold. The influence of self focusing is discussed, and evidence for beam distortion below the power threshold for complete self focusing is presented, confirming the theory of Marburger.

  10. Shock/vortex interaction and vortex-breakdown modes

    NASA Technical Reports Server (NTRS)

    Kandil, Osama A.; Kandil, H. A.; Liu, C. H.

    1992-01-01

    Computational simulation and study of shock/vortex interaction and vortex-breakdown modes are considered for bound (internal) and unbound (external) flow domains. The problem is formulated using the unsteady, compressible, full Navier-Stokes (NS) equations which are solved using an implicit, flux-difference splitting, finite-volume scheme. For the bound flow domain, a supersonic swirling flow is considered in a configured circular duct and the problem is solved for quasi-axisymmetric and three-dimensional flows. For the unbound domain, a supersonic swirling flow issued from a nozzle into a uniform supersonic flow of lower Mach number is considered for quasi-axisymmetric and three-dimensional flows. The results show several modes of breakdown; e.g., no-breakdown, transient single-bubble breakdown, transient multi-bubble breakdown, periodic multi-bubble multi-frequency breakdown and helical breakdown.

  11. Discriminating Dysarthria Type from Envelope Modulation Spectra

    ERIC Educational Resources Information Center

    Liss, Julie M.; LeGendre, Sue; Lotto, Andrew J.

    2010-01-01

    Purpose: Previous research demonstrated the ability of temporally based rhythm metrics to distinguish among dysarthrias with different prosodic deficit profiles (J. M. Liss et al., 2009). The authors examined whether comparable results could be obtained by an automated analysis of speech envelope modulation spectra (EMS), which quantifies the…

  12. Diffusive heat blanketing envelopes of neutron stars

    NASA Astrophysics Data System (ADS)

    Beznogov, M. V.; Potekhin, A. Y.; Yakovlev, D. G.

    2016-06-01

    We construct new models of outer heat blanketing envelopes of neutron stars composed of binary ion mixtures (H-He, He-C, C-Fe) in and out of diffusive equilibrium. To this aim, we generalize our previous work on diffusion of ions in isothermal gaseous or Coulomb liquid plasmas to handle non-isothermal systems. We calculate the relations between the effective surface temperature Ts and the temperature Tb at the bottom of heat blanketing envelopes (at a density ρb ˜ 108 - 1010 g cm-3) for diffusively equilibrated and non-equilibrated distributions of ion species at different masses ΔM of lighter ions in the envelope. Our principal result is that the Ts-Tb relations are fairly insensitive to detailed distribution of ion fractions over the envelope (diffusively equilibrated or not) and depend almost solely on ΔM. The obtained relations are approximated by analytic expressions which are convenient for modelling the evolution of neutron stars.

  13. Ozone Reductions Using Residential Building Envelopes

    SciTech Connect

    Walker, Iain S.; Sherman, Max; Nazaroff, William W.

    2009-02-01

    Ozone is an air pollutant with that can have significant health effects and a significant source of ozone in some regions of California is outdoor air. Because people spend the vast majority of their time indoors, reduction in indoor levels of ozone could lead to improved health for many California residents. Ozone is removed from indoor air by surface reactions and can also be filtered by building envelopes. The magnitude of the envelope impact depends on the specific building materials that the air flows over and the geometry of the air flow paths through the envelope that can be changes by mechanical ventilation operation. The 2008 Residential Building Standards in California include minimum requirements for mechanical ventilation by referencing ASHRAE Standard 62.2. This study examines the changes in indoor ozone depending on the mechanical ventilation system selected to meet these requirements. This study used detailed simulations of ventilation in a house to examine the impacts of different ventilation systems on indoor ozone concentrations. The simulation results showed that staying indoors reduces exposure to ozone by 80percent to 90percent, that exhaust ventilation systems lead to lower indoor ozone concentrations, that opening of windows should be avoided at times of high outdoor ozone, and that changing the time at which mechanical ventilation occurs has the ability to halve exposure to ozone. Future work should focus on the products of ozone reactions in the building envelope and the fate of these products with respect to indoor exposures.

  14. Ultraviolet Opacity and Fluorescence in Supernova Envelopes

    NASA Technical Reports Server (NTRS)

    Li, Hongwei; McCray, Richard

    1996-01-01

    By the time the expanding envelope of a Type 2 supernova becomes transparent in the optical continuum, most of the gamma-ray luminosity produced by radioactive Fe/Co/Ni clumps propagates into the hydrogen/helium envelope and is deposited there, if at all. The resulting fast electrons excite He 1 and H 1, the two- photon continua of which are the dominant internal sources of ultraviolet radiation. The UV radiation is blocked by scattering in thousands of resonance lines of metals and converted by fluorescence into optical and infrared emission lines that escape freely. We describe results of Monte Carlo calculations that simulate non-LTE scattering and fluorescence in more than five million allowed lines of Ca, Sc, Ti, V, Cr, Mn, Fe, Co, and Ni. For a model approximating conditions in the envelope of SN 1987A, the calculated emergent spectrum resembles the observed one. For the first 2 yr after explosion, the ultraviolet radiation (lambda less than or approximately equals 3000) is largely blocked and converted into a quasi continuum of many thousands of weak optical and infrared emission lines and some prominent emission features, such as the Ca 2 lambdalambda8600 triplet. Later, as the envelope cools and expands, it becomes more transparent, and an increasing fraction of the luminosity emerges in the UV band.

  15. The Story of the Red Envelopes.

    ERIC Educational Resources Information Center

    Lew, Gordon

    This is one of a series of elementary readers written in Cantonese and English and designed to familiarize children with the traditional major Chinese festivals celebrated by the Chinese in America. This booklet describes in narrative form the meaning of the red envelopes given with money gifts at Chinese New Year and other festivities. A page of…

  16. Tegument Assembly and Secondary Envelopment of Alphaherpesviruses.

    PubMed

    Owen, Danielle J; Crump, Colin M; Graham, Stephen C

    2015-09-01

    Alphaherpesviruses like herpes simplex virus are large DNA viruses characterized by their ability to establish lifelong latent infection in neurons. As for all herpesviruses, alphaherpesvirus virions contain a protein-rich layer called "tegument" that links the DNA-containing capsid to the glycoprotein-studded membrane envelope. Tegument proteins mediate a diverse range of functions during the virus lifecycle, including modulation of the host-cell environment immediately after entry, transport of virus capsids to the nucleus during infection, and wrapping of cytoplasmic capsids with membranes (secondary envelopment) during virion assembly. Eleven tegument proteins that are conserved across alphaherpesviruses have been implicated in the formation of the tegument layer or in secondary envelopment. Tegument is assembled via a dense network of interactions between tegument proteins, with the redundancy of these interactions making it challenging to determine the precise function of any specific tegument protein. However, recent studies have made great headway in defining the interactions between tegument proteins, conserved across alphaherpesviruses, which facilitate tegument assembly and secondary envelopment. We summarize these recent advances and review what remains to be learned about the molecular interactions required to assemble mature alphaherpesvirus virions following the release of capsids from infected cell nuclei. PMID:26393641

  17. Tegument Assembly and Secondary Envelopment of Alphaherpesviruses

    PubMed Central

    Owen, Danielle J.; Crump, Colin M.; Graham, Stephen C.

    2015-01-01

    Alphaherpesviruses like herpes simplex virus are large DNA viruses characterized by their ability to establish lifelong latent infection in neurons. As for all herpesviruses, alphaherpesvirus virions contain a protein-rich layer called “tegument” that links the DNA-containing capsid to the glycoprotein-studded membrane envelope. Tegument proteins mediate a diverse range of functions during the virus lifecycle, including modulation of the host-cell environment immediately after entry, transport of virus capsids to the nucleus during infection, and wrapping of cytoplasmic capsids with membranes (secondary envelopment) during virion assembly. Eleven tegument proteins that are conserved across alphaherpesviruses have been implicated in the formation of the tegument layer or in secondary envelopment. Tegument is assembled via a dense network of interactions between tegument proteins, with the redundancy of these interactions making it challenging to determine the precise function of any specific tegument protein. However, recent studies have made great headway in defining the interactions between tegument proteins, conserved across alphaherpesviruses, which facilitate tegument assembly and secondary envelopment. We summarize these recent advances and review what remains to be learned about the molecular interactions required to assemble mature alphaherpesvirus virions following the release of capsids from infected cell nuclei. PMID:26393641

  18. Validating predictions from climate envelope models

    USGS Publications Warehouse

    Watling, J.; Bucklin, D.; Speroterra, C.; Brandt, L.; Cabal, C.; Romañach, Stephanie S.; Mazzotti, Frank J.

    2013-01-01

    Climate envelope models are a potentially important conservation tool, but their ability to accurately forecast species’ distributional shifts using independent survey data has not been fully evaluated. We created climate envelope models for 12 species of North American breeding birds previously shown to have experienced poleward range shifts. For each species, we evaluated three different approaches to climate envelope modeling that differed in the way they treated climate-induced range expansion and contraction, using random forests and maximum entropy modeling algorithms. All models were calibrated using occurrence data from 1967–1971 (t1) and evaluated using occurrence data from 1998–2002 (t2). Model sensitivity (the ability to correctly classify species presences) was greater using the maximum entropy algorithm than the random forest algorithm. Although sensitivity did not differ significantly among approaches, for many species, sensitivity was maximized using a hybrid approach that assumed range expansion, but not contraction, in t2. Species for which the hybrid approach resulted in the greatest improvement in sensitivity have been reported from more land cover types than species for which there was little difference in sensitivity between hybrid and dynamic approaches, suggesting that habitat generalists may be buffered somewhat against climate-induced range contractions. Specificity (the ability to correctly classify species absences) was maximized using the random forest algorithm and was lowest using the hybrid approach. Overall, our results suggest cautious optimism for the use of climate envelope models to forecast range shifts, but also underscore the importance of considering non-climate drivers of species range limits. The use of alternative climate envelope models that make different assumptions about range expansion and contraction is a new and potentially useful way to help inform our understanding of climate change effects on species.

  19. Validating Predictions from Climate Envelope Models

    PubMed Central

    Watling, James I.; Bucklin, David N.; Speroterra, Carolina; Brandt, Laura A.; Mazzotti, Frank J.; Romañach, Stephanie S.

    2013-01-01

    Climate envelope models are a potentially important conservation tool, but their ability to accurately forecast species’ distributional shifts using independent survey data has not been fully evaluated. We created climate envelope models for 12 species of North American breeding birds previously shown to have experienced poleward range shifts. For each species, we evaluated three different approaches to climate envelope modeling that differed in the way they treated climate-induced range expansion and contraction, using random forests and maximum entropy modeling algorithms. All models were calibrated using occurrence data from 1967–1971 (t1) and evaluated using occurrence data from 1998–2002 (t2). Model sensitivity (the ability to correctly classify species presences) was greater using the maximum entropy algorithm than the random forest algorithm. Although sensitivity did not differ significantly among approaches, for many species, sensitivity was maximized using a hybrid approach that assumed range expansion, but not contraction, in t2. Species for which the hybrid approach resulted in the greatest improvement in sensitivity have been reported from more land cover types than species for which there was little difference in sensitivity between hybrid and dynamic approaches, suggesting that habitat generalists may be buffered somewhat against climate-induced range contractions. Specificity (the ability to correctly classify species absences) was maximized using the random forest algorithm and was lowest using the hybrid approach. Overall, our results suggest cautious optimism for the use of climate envelope models to forecast range shifts, but also underscore the importance of considering non-climate drivers of species range limits. The use of alternative climate envelope models that make different assumptions about range expansion and contraction is a new and potentially useful way to help inform our understanding of climate change effects on species. PMID

  20. The envelope-based cyclic periodogram

    NASA Astrophysics Data System (ADS)

    Borghesani, P.

    2015-06-01

    Cyclostationary analysis has proven effective in identifying signal components for diagnostic purposes. A key descriptor in this framework is the cyclic power spectrum, traditionally estimated by the averaged cyclic periodogram and the smoothed cyclic periodogram. A lengthy debate about the best estimator finally found a solution in a cornerstone work by Antoni, who proposed a unified form for the two families, thus allowing a detailed statistical study of their properties. Since then, the focus of cyclostationary research has shifted towards algorithms, in terms of computational efficiency and simplicity of implementation. Traditional algorithms have proven computationally inefficient and the sophisticated "cyclostationary" definition of these estimators slowed their spread in the industry. The only attempt to increase the computational efficiency of cyclostationary estimators is represented by the cyclic modulation spectrum. This indicator exploits the relationship between cyclostationarity and envelope analysis. The link with envelope analysis allows a leap in computational efficiency and provides a "way in" for the understanding by industrial engineers. However, the new estimator lies outside the unified form described above and an unbiased version of the indicator has not been proposed. This paper will therefore extend the analysis of envelope-based estimators of the cyclic spectrum, proposing a new approach to include them in the unified form of cyclostationary estimators. This will enable the definition of a new envelope-based algorithm and the detailed analysis of the properties of the cyclic modulation spectrum. The computational efficiency of envelope-based algorithms will be also discussed quantitatively for the first time in comparison with the averaged cyclic periodogram. Finally, the algorithms will be validated with numerical and experimental examples.

  1. RF BREAKDOWN STUDIES USING PRESSURIZED CAVITIES

    SciTech Connect

    Johnson, Rolland

    2014-09-21

    Many present and future particle accelerators are limited by the maximum electric gradient and peak surface fields that can be realized in RF cavities. Despite considerable effort, a comprehensive theory of RF breakdown has not been achieved and mitigation techniques to improve practical maximum accelerating gradients have had only limited success. Part of the problem is that RF breakdown in an evacuated cavity involves a complex mixture of effects, which include the geometry, metallurgy, and surface preparation of the accelerating structures and the make-up and pressure of the residual gas in which plasmas form. Studies showed that high gradients can be achieved quickly in 805 MHz RF cavities pressurized with dense hydrogen gas, as needed for muon cooling channels, without the need for long conditioning times, even in the presence of strong external magnetic fields. This positive result was expected because the dense gas can practically eliminate dark currents and multipacting. In this project we used this high pressure technique to suppress effects of residual vacuum and geometry that are found in evacuated cavities in order to isolate and study the role of the metallic surfaces in RF cavity breakdown as a function of magnetic field, frequency, and surface preparation. One of the interesting and useful outcomes of this project was the unanticipated collaborations with LANL and Fermilab that led to new insights as to the operation of evacuated normal-conducting RF cavities in high external magnetic fields. Other accomplishments included: (1) RF breakdown experiments to test the effects of SF6 dopant in H2 and He gases with Sn, Al, and Cu electrodes were carried out in an 805 MHz cavity and compared to calculations and computer simulations. The heavy corrosion caused by the SF6 components led to the suggestion that a small admixture of oxygen, instead of SF6, to the hydrogen would allow the same advantages without the corrosion in a practical muon beam line. (2) A

  2. The Psp system of Mycobacterium tuberculosis integrates envelope stress sensing and envelope preserving functions

    PubMed Central

    Datta, Pratik; Ravi, Janani; Guerrini, Valentina; Chauhan, Rinki; Neiditch, Matthew B.; Shell, Scarlet S.; Fortune, Sarah M.; Hancioglu, Baris; Igoshin, Oleg; Gennaro, Maria Laura

    2015-01-01

    The bacterial envelope integrates essential stress-sensing and adaptive functions; thus, envelope-preserving functions are important for survival. In Gram-negative bacteria, envelope integrity during stress is maintained by the multi-gene Psp response. Mycobacterium tuberculosis was thought to lack the Psp system, since it encodes only pspA and no other psp ortholog. Intriguingly, pspA maps downstream from clgR, which encodes a transcription factor regulated by the MprAB-σE envelope-stress-signaling system. clgR inactivation lowered ATP concentration during stress and protonophore treatment-induced clgR-pspA expression, suggesting that these genes express Psp-like functions. We identified a four-gene set -- clgR, pspA (rv2744c), rv2743c, rv2742c – that is regulated by clgR and in turn regulates ClgR activity. Regulatory and protein-protein interactions within the set and a requirement of the four genes for functions associated with envelope integrity and surface-stress tolerance indicate that a Psp-like system has evolved in mycobacteria. Among Actinobacteria, the four-gene module occurred only in tuberculous mycobacteria and was required for intra-macrophage growth, suggesting links between its function and mycobacterial virulence. Additionally, the four-gene module was required for MprAB-σE stress-signaling activity. The positive feedback between envelope-stress-sensing and envelope-preserving functions allows sustained responses to multiple, envelope-perturbing signals during chronic infection, making the system uniquely suited to tuberculosis pathogenesis. PMID:25899163

  3. Phytochrome regulates GTP-binding protein activity in the envelope of pea nuclei

    NASA Technical Reports Server (NTRS)

    Clark, G. B.; Memon, A. R.; Thompson, G. A. Jr; Roux, S. J.

    1993-01-01

    Three GTP-binding proteins with apparent molecular masses of 27, 28 and 30 kDa have been detected in isolated nuclei of etiolated pea plumules. After LDS-PAGE and transfer to nitrocellulose these proteins bind [32P]GTP in the presence of excess ATP, suggesting that they are monomeric G proteins. When nuclei are disrupted, three proteins co-purify with the nuclear envelope fraction and are highly enriched in this fraction. The level of [32P]GTP-binding for all three protein bands is significantly increased when harvested pea plumules are irradiated by red light, and this effect is reversed by far-red light. The results indicate that GTP-binding activity associated with the nuclear envelope of plant cells is photoreversibly regulated by the pigment phytochrome.

  4. Solar Effective Envelope Design Advisor (SEEDA)

    NASA Astrophysics Data System (ADS)

    Mahaek, Ekkachai

    The lack of effort by mainstream architects in integrating energy-efficient strategies in architectural designing is due to the complexity in a building's energy conscious concepts and theories, the difficulties to visualize and quantify energy consumption, and the late implementing of energy consumption analysis in the conventional design process. This task would be accomplishing by a building system's engineer where results might be determined only after the basic architectural design has been completed. An effective simple tool and method should then be available to assist architects in building's energy-efficient designing at the beginning of the design. The building's energy consumption is directly and mainly influenced by the relationship of the sun, site, and its building configuration. The solar radiations will first impact on the building's envelope, which will have a direct effect on the amount of energy a building will consume. If an architect can define or map the intensity of solar energy on the site's buildable volume, and use this information to determine the levels of solar insolation, a more energy efficient building form can be proposed. This research hypothesis has shared the fundamental techniques of the Solar Envelope projection by Professor Ralph Knowles [Knowles, 1981] of the University of Southern California. However a different approach is taken by including the influence of regional restrictions and the surrounding buildings' shadows when projecting of solar volumes and solar envelope. The research methodology will discuss the development of a computer-based approach to develop a three-dimensional architectural form based on an insolation map related to the design site. The prototype computer program is referred as the Solar Effective Envelope Design Advisor (SEEDA). The solar insolation volume of the site is determined by integrating three types of computer-generated models include the Buildable Volume model based on design constraints

  5. Analysis of Building Envelope Construction in 2003 CBECS

    SciTech Connect

    Winiarski, David W.; Halverson, Mark A.; Jiang, Wei

    2007-06-01

    The purpose of this analysis is to determine "typical" building envelope characteristics for buildings built after 1980. We address three envelope components in this paper - roofs, walls, and window area. These typical building envelope characteristics were used in the development of DOE’s Reference Buildings .

  6. Human skeletal muscle protein breakdown during spaceflight

    NASA Technical Reports Server (NTRS)

    Stein, T. P.; Schluter, M. D.

    1997-01-01

    Human spaceflight is associated with a loss of body protein. Excretion of 3-methylhistidine (3-MH) in the urine is a useful measurement of myofibrillar protein breakdown. Bed rest, particularly with 6 degrees head-down tilt, is an accepted ground-based model for human spaceflight. The objectives of this report were to compare 3-MH excretion from two Life Sciences shuttle missions (duration 9.5 and 15 days, n = 9) and from 17 days of bed rest (n = 7) with 6 degrees head-down tilt. The bed rest study was designed to mimic an actual Life Sciences spaceflight and so incorporated an extensive battery of physiological tests focused on the musculoskeletal system. Results showed that nitrogen retention, based on excretion of nitrogen in the urine, was reduced during both bed rest [from 22 +/- 1 to 1 +/- 5 mg N x kg(-1) x day(-1) (n = 7; P < 0.05)] and spaceflight [from 57 +/- 9 to 19 +/- 3 mg N x kg(-1) x day(-1) (n = 9; P < 0.05)]. 3-MH excretion was unchanged with either bed rest [pre-bed rest 5.30 +/- 0.29 vs. bed rest 5.71 +/- 0.30 micromol 3-MH x kg(-1) x day(-1), n = 7; P = not significant (NS)] or spaceflight [preflight 4.98 +/- 0.37 vs. 4.59 +/- 0.39 micromol 3-MH x kg(-1) x day(-1) in-flight, n = 9; P = NS]. We conclude that 1) 3-MH excretion was unaffected by spaceflight on the shuttle or with bed rest plus exercise, and 2) because protein breakdown (elevated 3-MH) was increased on Skylab but not on the shuttle, it follows that muscle protein breakdown is not an inevitable consequence of spaceflight.

  7. Nanolaminates: increasing dielectric breakdown strength of composites.

    PubMed

    Fillery, Scott P; Koerner, Hilmar; Drummy, Lawrence; Dunkerley, Erik; Durstock, Michael F; Schmidt, Daniel F; Vaia, Richard A

    2012-03-01

    Processable, low-cost, high-performance hybrid dielectrics are enablers for a vast array of green technologies, including high-temperature electrical insulation and pulsed power capacitors for all-electric transportation vehicles. Maximizing the dielectric breakdown field (E(BD)), in conjunction with minimization of leakage current, directly impacts system performance because of the field's quadratic relationship with electrostatic energy storage density. On the basis of the extreme internal interfacial area and ultrafine morphology, polymer-inorganic nanocomposites (PNCs) have demonstrated modest increases in E(BD) at very low inorganic loadings, but because of insufficient control of the hierarchal morphology of the blend, have yielded a precipitous decline in E(BD) at intermediate and high inorganic volume fractions. Here in, we demonstrate that E(BD) can be increased up to these intermediate inorganic volume fractions by creating uniform one-dimensional nanocomposites (nanolaminates) rather than blends of spherical inorganic nanoparticles and polymers. Free standing nanolaminates of highly aligned and dispersed montmorillonite in polyvinyl butyral exhibited enhancements in E(BD) up to 30 vol % inorganic (70 wt % organically modified montmorillonite). These relative enhancements extend up to five times the inorganic fraction observed for random nanoparticle dispersions, and are anywhere from two to four times greater than observed at comparable volume fraction of nanoparticles. The breakdown characteristics of this model system suggested a trade-off between increased path tortuosity and polymer-deficient structural defects. This implies that an idealized PNC morphology to retard the breakdown cascade perpendicular to the electrodes will occur at intermediate volume fractions and resemble a discotic nematic phase where highly aligned, high-aspect ratio nanometer thick plates are uniformly surrounded by nanoscopic regions of polymer.

  8. Theoretical and experimental investigation of vortex breakdown

    NASA Technical Reports Server (NTRS)

    Krause, E.

    1986-01-01

    The slender-vortex approximation was analyzed for incompressible and compressible flow. First the equations of motion were reduced in an order of magnitude analysis. Then compatibility conditions were formulated for the inflow conditions. Thereafter finite-difference-solutions were constructed for incompressible and compressible flow. Finally it was shown that these solutions can be used to describe the flow in slender vortices. The analysis of the breakdown process must, however, be excluded, since its upstream influence cannot be predicted with the slender vortex approximation. The investigaton of this problem is left for future work.

  9. Breakdown criteria for nonvacuum Einstein equations

    NASA Astrophysics Data System (ADS)

    Shao, Arick

    We generalize a recent "breakdown criterion" result of S. Klainerman and I. Rodnianski, which states roughly that an Einstein vacuum spacetime, given as a CMC foliation, can be extended if the second fundamental form and the derivative of the lapse of the foliation are uniformly bounded. We adapt this theorem and its proof to Einstein-scalar and Einstein-Maxwell spacetimes. In particular, we deal with additional issues resulting from nontrivial Ricci curvature and the coupling between the Einstein and the field equations. The results we prove can be directly extended to Einstein-Klein-Gordon and Einstein-Yang-Mills spacetimes.

  10. Laser-induced breakdown spectroscopy in Asia

    NASA Astrophysics Data System (ADS)

    Wang, Zhen-Zhen; Deguchi, Yoshihiro; Zhang, Zhen-Zhen; Wang, Zhe; Zeng, Xiao-Yan; Yan, Jun-Jie

    2016-12-01

    Laser-induced breakdown spectroscopy (LIBS) is an analytical detection technique based on atomic emission spectroscopy to measure the elemental composition. LIBS has been extensively studied and developed due to the non-contact, fast response, high sensitivity, real-time and multi-elemental detection features. The development and applications of LIBS technique in Asia are summarized and discussed in this review paper. The researchers in Asia work on different aspects of the LIBS study in fundamentals, data processing and modeling, applications and instrumentations. According to the current research status, the challenges, opportunities and further development of LIBS technique in Asia are also evaluated to promote LIBS research and its applications.

  11. Kinetic theory of runaway air-breakdown

    SciTech Connect

    Roussel-Dupre, R.A.; Gurevich, A.V.; Tunnell, T.; Milikh, G.M.

    1993-09-01

    The kinetic theory for a new air breakdown mechanism advanced in a previous paper is developed. The relevant form of the Boltzmann equation is derived and the particle orbits in both velocity space and configuration space are computed. A numerical solution of the Boltzmann equation, assuming a spatially uniform electric field, is obtained and the temporal evolution of the electron velocity distribution function is described. The results of our analysis are used to estimate the magnitude of potential x-ray emissions from discharges in thunderstorms.

  12. Kinetic theory of runaway air breakdown

    SciTech Connect

    Roussel-Dupre, R.A. ); Gurevich, A.V. ); Tunnell, T. ); Milikh, G.M. )

    1994-03-01

    The kinetic theory for an air breakdown mechanism advanced in a previous paper [Phys. Lett. A 165, 463 (1992)] is developed. The relevant form of the Boltzmann equation is derived and the particle orbits in both velocity space and configuration space are computed. A numerical solution of the Boltzmann equation, assuming a spatially uniform electric field, is obtained and the temporal evolution of the electron velocity distribution function is described. The results of our analysis are used to estimate the magnitude of potential x-ray emissions from discharges in thunderstorms.

  13. Transient Wave Envelope Elements for Wave Problems

    NASA Astrophysics Data System (ADS)

    Astley, R. J.

    1996-04-01

    A novel family of infinite wave envelope elements is described which can be used in conjunction with conventional finite elements to model the transient wave equation in unbounded regions. The elements are obtained by applying an inverse Fourier transformation to a mapped wave envelope formulation in the frequency domain. The discrete transient equations obtained in this way can be applied to two-dimensional and three-dimensional problems without restriction, being valid over a full range of excitation frequencies. The effectiveness and accuracy of the method is demonstrated in application to simple test cases which involve the calculation of transient sound fields generated by pulsating spheres and cylinders excited from rest in an unbounded region. Test solutions are compared to analytic solutions and to finite element solutions obtained by using large computational grids which extend beyond the region influenced by the transient disturbance.

  14. Development of High Specific Strength Envelope Materials

    NASA Astrophysics Data System (ADS)

    Komatsu, Keiji; Sano, Masa-Aki; Kakuta, Yoshiaki

    Progress in materials technology has produced a much more durable synthetic fabric envelope for the non-rigid airship. Flexible materials are required to form airship envelopes, ballonets, load curtains, gas bags and covering rigid structures. Polybenzoxazole fiber (Zylon) and polyalirate fiber (Vectran) show high specific tensile strength, so that we developed membrane using these high specific tensile strength fibers as a load carrier. The main material developed is a Zylon or Vectran load carrier sealed internally with a polyurethane bonded inner gas retention film (EVOH). The external surface provides weather protecting with, for instance, a titanium oxide integrated polyurethane or Tedlar film. The mechanical test results show that tensile strength 1,000 N/cm is attained with weight less than 230g/m2. In addition to the mechanical properties, temperature dependence of the joint strength and solar absorptivity and emissivity of the surface are measured. 

  15. Discontinuous envelope function in semiconductor heterostructures

    NASA Astrophysics Data System (ADS)

    Drouhin, Henri-Jean; Bottegoni, Federico; Nguyen, T. L. Hoai; Wegrowe, Jean-Eric; Fishman, Guy

    2013-09-01

    Based on a proper definition of the current operators for non-quadratic Hamiltonians, we derive the expression for the transport current which involves the derivative of the imaginary part of the free-electron current, highlighting peculiarities of the extra terms. The expression of the probability current, when Spin-Orbit Interaction (SOI) is taken into account, requires a reformulation of the boudary conditions. This is especially important for tunnel heterojunctions made of non-centrosymmetric semiconductors. Therefore, we consider a model case: tunneling of conduction electrons through a [110]-oriented GaAs barrier. The new boundary conditions are reduced to two set of equations: the first one expresses the discontinuity of the envelope function at the interface while the other one expresses the discontinuity of the derivative of the envelope function.

  16. Uses and misuses of bioclimatic envelope modeling.

    PubMed

    Araújo, Miguel B; Peterson, A Townsend

    2012-07-01

    Bioclimatic envelope models use associations between aspects of climate and species' occurrences to estimate the conditions that are suitable to maintain viable populations. Once bioclimatic envelopes are characterized, they can be applied to a variety of questions in ecology, evolution, and conservation. However, some have questioned the usefulness of these models, because they may be based on implausible assumptions or may be contradicted by empirical evidence. We review these areas of contention, and suggest that criticism has often been misplaced, resulting from confusion between what the models actually deliver and what users wish that they would express. Although improvements in data and methods will have some effect, the usefulness of these models is contingent on their appropriate use, and they will improve mainly via better awareness of their conceptual basis, strengths, and limitations.

  17. Development of laser induced breakdown spectroscopy instrumentatin for safeguards applications

    SciTech Connect

    Barefield Il, James E; Clegg, Samuel M; Le, Loan A; Lopez, Leon N

    2010-01-01

    In September 2006, a Technical Meeting on Application of Laser Spectrometry Techniques in IAEA Safeguards was held at IAEA headquarters (HQ). One of the principal recommendations from this meeting was the need to 'pursue the development of novel complementary access instrumentation based on laser induced breakdown spectroscopy (LIBS) for the detection of gaseous and solid signatures and indicators of nuclear fuel cycle processes and associated materials.' Pursuant to this recommendation the Department of Safeguards (SG) under the Division of Technical Support (SGTS) convened the Experts and Users Advisory Meeting on Laser Induced Breakdown Spectroscopy (LIBS) for Safeguards Applications. This meeting was held at IAEA HQ from July 7-11,2008 and hosted by the Novel Technologies Unit (NTU). The meeting was attended by 12 LIBS experts from the Czech Republic, the European Commission, France, the Republic of Korea, the United States of America, Germany, the United Kingdom of Great Britain, Canada, and Northern Ireland. After a presentation of the needs of the IAEA inspectors, the LIBS experts were in agreement that needs as presented could be partially or fully fulfilled using LIBS instrumentation. The needs of the IAEA inspectors were grouped in the following broad categories: (1) Improvements to in-field measurements/environmental sampling; (2) Monitoring status of activity in a Hot Cell; (3) Verifying status of activity at a declared facility via process monitoring; and (4) Need for pre-screening of environmental samples before analysis. Under the Department of Energy/National Nuclear Security Administration (DOE/NNSA) Next Generation Safeguards Initiative (NGSI) Los Alamos National Laboratory is exploring three potential applications of LIBS for international safeguards. As part of this work, we are developing: (1) a user-friendly man-portable LIBS system to characterize samples across a wide range of elements in the periodic table from hydrogen up to heavy elements

  18. Digital image envelope: method and evaluation

    NASA Astrophysics Data System (ADS)

    Huang, H. K.; Cao, Fei; Zhou, Michael Z.; Mogel, Greg T.; Liu, Brent J.; Zhou, Xiaoqiang

    2003-05-01

    Health data security, characterized in terms of data privacy, authenticity, and integrity, is a vital issue when digital images and other patient information are transmitted through public networks in telehealth applications such as teleradiology. Mandates for ensuring health data security have been extensively discussed (for example The Health Insurance Portability and Accountability Act, HIPAA) and health informatics guidelines (such as the DICOM standard) are beginning to focus on issues of data continue to be published by organizing bodies in healthcare; however, there has not been a systematic method developed to ensure data security in medical imaging Because data privacy and authenticity are often managed primarily with firewall and password protection, we have focused our research and development on data integrity. We have developed a systematic method of ensuring medical image data integrity across public networks using the concept of the digital envelope. When a medical image is generated regardless of the modality, three processes are performed: the image signature is obtained, the DICOM image header is encrypted, and a digital envelope is formed by combining the signature and the encrypted header. The envelope is encrypted and embedded in the original image. This assures the security of both the image and the patient ID. The embedded image is encrypted again and transmitted across the network. The reverse process is performed at the receiving site. The result is two digital signatures, one from the original image before transmission, and second from the image after transmission. If the signatures are identical, there has been no alteration of the image. This paper concentrates in the method and evaluation of the digital image envelope.

  19. Time-dependent MOS breakdown. [of Na contaminated capacitors

    NASA Technical Reports Server (NTRS)

    Li, S. P.; Bates, E. T.; Maserjian, J.

    1976-01-01

    A general model for time-dependent breakdown in metal-oxide-silicon (MOS) structures is developed and related to experimental measurements on samples deliberately contaminated with Na. A statistical method is used for measuring the breakdown probability as a function of log time and applied field. It is shown that three time regions of breakdown can be explained respectively in terms of silicon surface defects, ion emission from the metal interface, and lateral ion diffusion at the silicon interface.

  20. The cell envelope glycoconjugates of Mycobacterium tuberculosis

    PubMed Central

    Angala, Shiva Kumar; Belardinelli, Juan Manuel; Huc-Claustre, Emilie; Wheat, William H.; Jackson, Mary

    2015-01-01

    Tuberculosis (TB) remains the second most common cause of death due to a single infectious agent. The cell envelope of Mycobacterium tuberculosis (Mtb), the causative agent of the disease in humans, is a source of unique glycoconjugates and the most distinctive feature of the biology of this organism. It is the basis of much of Mtb pathogenesis and one of the major causes of its intrinsic resistance to chemotherapeutic agents. At the same time, the unique structures of Mtb cell envelope glycoconjugates, their antigenicity and essentiality for mycobacterial growth provide opportunities for drug, vaccine, diagnostic and biomarker development, as clearly illustrated by recent advances in all of these translational aspects. This review focuses on our current understanding of the structure and biogenesis of Mtb glycoconjugates with particular emphasis on one of most intriguing and least understood aspect of the physiology of mycobacteria: the translocation of these complex macromolecules across the different layers of the cell envelope. It further reviews the rather impressive progress made in the last ten years in the discovery and development of novel inhibitors targeting their biogenesis. PMID:24915502

  1. Chimeric Measles Viruses with a Foreign Envelope

    PubMed Central

    Spielhofer, Pius; Bächi, Thomas; Fehr, Thomas; Christiansen, Gudrun; Cattaneo, Roberto; Kaelin, Karin; Billeter, Martin A.; Naim, Hussein Y.

    1998-01-01

    Measles virus (MV) and vesicular stomatitis virus (VSV) are both members of the Mononegavirales but are only distantly related. We generated two genetically stable chimeric viruses. In MGV, the reading frames of the MV envelope glycoproteins H and F were substituted by a single reading frame encoding the VSV G glycoprotein; MG/FV is similar but encodes a G/F hybrid in which the VSV G cytoplasmic tail was replaced by that of MV F. In contrast to MG/FV, MGV virions do not contain the MV matrix (M) protein. This demonstrates that virus assembly is possible in the absence of M; conversely, the cytoplasmic domain of F allows incorporation of M and enhances assembly. The formation of chimeric viruses was substantially delayed and the titers obtained were reduced about 50-fold in comparison to standard MV. In the novel chimeras, transcription and replication are mediated by the MV ribonucleoproteins but the envelope glycoproteins dictate the host range. Mice immunized with the chimeric viruses were protected against lethal doses of wild-type VSV. These findings suggest that it is feasible to construct MV variants bearing a variety of different envelopes for use as vaccines or for gene therapeutic purposes. PMID:9499071

  2. Fusion of Enveloped Viruses in Endosomes.

    PubMed

    White, Judith M; Whittaker, Gary R

    2016-06-01

    Ari Helenius launched the field of enveloped virus fusion in endosomes with a seminal paper in the Journal of Cell Biology in 1980. In the intervening years, a great deal has been learned about the structures and mechanisms of viral membrane fusion proteins as well as about the endosomes in which different enveloped viruses fuse and the endosomal cues that trigger fusion. We now recognize three classes of viral membrane fusion proteins based on structural criteria and four mechanisms of fusion triggering. After reviewing general features of viral membrane fusion proteins and viral fusion in endosomes, we delve into three characterized mechanisms for viral fusion triggering in endosomes: by low pH, by receptor binding plus low pH and by receptor binding plus the action of a protease. We end with a discussion of viruses that may employ novel endosomal fusion-triggering mechanisms. A key take-home message is that enveloped viruses that enter cells by fusing in endosomes traverse the endocytic pathway until they reach an endosome that has all of the environmental conditions (pH, proteases, ions, intracellular receptors and lipid composition) to (if needed) prime and (in all cases) trigger the fusion protein and to support membrane fusion.

  3. Effect of droplet-induced breakdown on CARS temperature measurements

    SciTech Connect

    Dunn-Rankin, D. ); Switzer, G.L. ); Obringer, C.A.; Jackson, T. )

    1990-07-20

    This research examines the potential for coherent anti-Stokes Raman scattering (CARS) to rovide reliable gas temperature measurements in the presence of liquid droplets. The droplets cause dielectric breakdown by focusing the CARS laser beams. This breakdown produces a plasma that can disrupt or obscure the CARS signal. Specifically, we examine the influence of laser induced breakdown on the CARS signal, and we determine the importance of droplet position relative to the CARS focal volume and droplet concentration on the reliability of CARS temperature measurements in droplet-laden flows. In addition, we propose a reliable data reduction procedure to minimize the disruptive influence of laser induced breakdown on CARS temperature.

  4. Laser radiation attenuation by sparks of optical breakdown

    NASA Astrophysics Data System (ADS)

    Budnik, A. P.; Semenov, L. P.; Skripkin, A. M.; Volkovitskii, O. A.

    1989-06-01

    A breakdown generated by laser radiation in a gas contaminated by aerosol particles is known to occur at much lower radiation intensities than in case of pure gases. Laser radiation is heavily attenuated by sparks of plasma formed at breakdowns. Energy loss estimation is important at radiation propagation in the atmosphere and in laser resonators. The breakdown phenomenon may be used in diagnostics of the atmospheric aerosol contamination events. The report presents experimental data on the influence of aerosol size distribution and concentration on optical breakdown generation and other results.

  5. Laser Radiation Attenuation By Sparks Of Optical Breakdown

    NASA Astrophysics Data System (ADS)

    Budnik, A. P.; Semenov, L. P.; Skripkin, A. M.; Volkovitsky, O. A.

    1990-01-01

    A breakdown generated by laser radiation in a gas contaminated by aerosol particles is known to occur at much lower radiation intensities than in case of pure gases, Laser radiation is heavily attenuated by sparks of plasma formed at breakdowns. Energy loss estimation is important at radiation propagation in the atmosphere and in laser resonators. The breakdown phenomenon may be used in diagnostics of the atmospheric aerosol contamination events. The report presents experimental data on the influence of aerosol size distribution and concentration on optical breakdown generation and other results.

  6. Simulations of avalanche breakdown statistics: probability and timing

    NASA Astrophysics Data System (ADS)

    Ng, Jo Shien; Tan, Chee Hing; David, John P. R.

    2010-04-01

    Important avalanche breakdown statistics for Single Photon Avalanche Diodes (SPADs), such as avalanche breakdown probability, dark count rate, and the distribution of time taken to reach breakdown (providing mean time to breakdown and jitter), were simulated. These simulations enable unambiguous studies on effects of avalanche region width, ionization coefficient ratio and carrier dead space on the avalanche statistics, which are the fundamental limits of the SPADs. The effects of quenching resistor/circuit have been ignored. Due to competing effects between dead spaces, which are significant in modern SPADs with narrow avalanche regions, and converging ionization coefficients, the breakdown probability versus overbias characteristics from different avalanche region widths are fairly close to each other. Concerning avalanche breakdown timing at given value of breakdown probability, using avalanche material with similar ionization coefficients yields fast avalanche breakdowns with small timing jitter (albeit higher operating field), compared to material with dissimilar ionization coefficients. This is the opposite requirement for abrupt breakdown probability versus overbias characteristics. In addition, by taking band-to-band tunneling current (dark carriers) into account, minimum avalanche region width for practical SPADs was found to be 0.3 and 0.2 μm, for InP and InAlAs, respectively.

  7. Pre-breakdown evaluation of gas discharge mechanisms in microgaps

    SciTech Connect

    Semnani, Abbas; Peroulis, Dimitrios; Venkattraman, Ayyaswamy; Alexeenko, Alina A.

    2013-04-29

    The individual contributions of various gas discharge mechanisms to total pre-breakdown current in microgaps are quantified numerically. The variation of contributions of field emission and secondary electron emission with increasing electric field shows contrasting behavior even for a given gap size. The total current near breakdown decreases rapidly with gap size indicating that microscale discharges operate in a high-current, low-voltage regime. This study provides the first such analysis of breakdown mechanisms and aids in the formulation of physics-based theories for microscale breakdown.

  8. Locating Preliminary Breakdown Pulses in Lightning Flashes

    NASA Astrophysics Data System (ADS)

    Karunarathne, S.; Marshall, T.; Stolzenburg, M.

    2011-12-01

    Lightning flashes often begin with a series of bipolar pulses, 1-5 us in width, called initial breakdown pulses or characteristic pulses. In 2010 we showed that these pulses can be located (find x, y, z, t) using Time of arrival method (TOA) [Koshak and Solakiewicz, JGR, 1996]. Electric field change data was obtained at the NASA/Kennedy Space Center (KSC) during the summer of 2010 at 5 stations with a band width of 0-0.5MHz and time accuracy of 1us. We concluded that in order to increase the accuracy positions; time accuracy, band width and number of stations should be increased. In summer of 2011, we placed electric field change meters with band width of 0-5Mhz and time accuracy of 0.1us at the KSC. We have doubled the number of stations (10 stations). We use TOA technique with different algorithm to locate positions of beginning pulses with greater accuracy. The locations will be compared to locations of VHF lightning sources made with the KSC LDAR2 system (which has a center frequency of 63 MHz and a bandwidth of 6 MHz). A monte-carlo method will be used to calculate the error of the locations. A statistical comparison between our TOA positions and LDAR2 positions will be presented along with possible physical connections between the preliminary breakdown pulses, the LDAD2 sources, and the developing lightning leader.

  9. Functions of Carotenoid Metabolites and Breakdown Products

    NASA Astrophysics Data System (ADS)

    Britton, George

    It is not only intact carotenoids but also fragments of carotenoid molecules that have important natural functions and actions. The electron-rich polyene chain of the carotenoids is very susceptible to oxidative breakdown, which may be enzymic or non-enzymic. Central cleavage gives C20 compounds, retinoids, as described in Chapter 16. Cleavage at other positions gives smaller fragments, notably C10, C13 and C15 compounds that retain the carotenoid end group. The formation of these is described in Chapter 17 and in Volume 3, Chapter 4. Oxidative breakdown can also take place during storage, processing and curing of plant material, and the products contribute to the desired aroma/flavour properties of, for example, tea, wine and tobacco. The importance of vitamin A (C20) in animals is well known. Vitamin A deficiency is still a major concern in many parts of the world. It can lead to blindness and serious ill-health or death, especially in young children. Volatile smaller carotenoid fragments (`norisoprenoids') are widespread scent/flavour compounds in plants.

  10. Laser Induced Breakdown Spectroscopy of Metals

    NASA Astrophysics Data System (ADS)

    Palmer, Andria; Lawhead, Carlos; Ujj, Laszlo

    2015-03-01

    Laser Induced Breakdown Spectroscopy (LIBS) is a very practical spectroscopy to determine the chemical composition of materials. Recent technical developments resulted in equipment used on the MARS Rover by NASA. It is capable of measuring the emission spectra of laser induced plasma created by energetic laser pulses focused on the sample (rocks, metals, etc.). We have develop a Laser Induced Breakdown Spectroscopy setup and investigated the necessary experimental and methodological challenges needed to make such material identification measurements. 355 and 532 nm laser pulses with 5 ns temporal duration was used to generate micro-plasma from which compositions can be determined based on known elemental and molecular emission intensities and wavelengths. The performance of LIBS depends on several parameters including laser wavelength, pulse energy, pulse duration, time interval of observation, geometrical configuration of collecting optics, and the properties of ambient medium. Spectra recorded from alloys (e.g. US penny coin) and pure metals will be presented. Special thanks for the financial support of the Office of Undergraduate Research of UWF.

  11. Preparation and Characterization of Envelope Membranes from Nongreen Plastids

    PubMed Central

    Alban, Claude; Joyard, Jacques; Douce, Roland

    1988-01-01

    We have developed a reliable procedure for the purification of envelope membranes from cauliflower (Brassica oleracea L.) bud plastids and sycamore (Acer pseudoplatanus L.) cell amyloplasts. After disruption of purified intact plastids, separation of envelope membranes was achieved by centrifugation on a linear sucrose gradient. A membrane fraction, having a density of 1.122 grams per cubic centimeter and containing carotenoids, was identified as the plastid envelope by the presence of monogalactosyldiacylglycerol synthase. Using antibodies raised against spinach chloroplast envelope polypeptides E24 and E30, we have demonstrated that both the outer and the inner envelope membranes were present in this envelope fraction. The major polypeptide in the envelope fractions from sycamore and cauliflower plastids was identified immunologically as the phosphate translocator. In the envelope membranes from cauliflower and sycamore plastids, the major glycerolipids were monogalactosyldiacylglycerol, digalactosyldiacylglycerol, and phosphatidylcholine. Purified envelope membranes from cauliflower bud plastids and sycamore amyloplasts also contained a galactolipid:galactolipid galactosyltransferase, enzymes for phosphatidic acid and diacylglycerol biosynthesis, acyl-coenzyme A thioesterase, and acyl-coenzyme A synthetase. These results demonstrate that envelope membranes from nongreen plastids present a high level of homology with chloroplasts envelope membranes. Images Fig. 4 Fig. 5 Fig. 6 Fig. 7 PMID:16666372

  12. Antiviral Activity of Graphene–Silver Nanocomposites against Non-Enveloped and Enveloped Viruses

    PubMed Central

    Chen, Yi-Ning; Hsueh, Yi-Huang; Hsieh, Chien-Te; Tzou, Dong-Ying; Chang, Pai-Ling

    2016-01-01

    The discovery of novel antiviral materials is important because many infectious diseases are caused by viruses. Silver nanoparticles have demonstrated strong antiviral activity, and graphene is a potential antimicrobial material due to its large surface area, high carrier mobility, and biocompatibility. No studies on the antiviral activity of nanomaterials on non-enveloped viruses have been reported. To investigate the antiviral activity of graphene oxide (GO) sheets and GO sheets with silver particles (GO-Ag) against enveloped and non-enveloped viruses, feline coronavirus (FCoV) with an envelope and infectious bursal disease virus (IBDV) without an envelope were chosen. The morphology and sizes of GO and GO-Ag were characterized by transmission, scanning electron microscopy, and X-ray diffraction. A virus inhibition assay was used to identify the antiviral activity of GO and GO-Ag. Go-Ag inhibited 25% of infection by FCoV and 23% by IBDV, whereas GO only inhibited 16% of infection by FCoV but showed no antiviral activity against the infection by IBDV. Further application of GO and GO-Ag can be considered for personal protection equipment to decrease the transmission of viruses. PMID:27104546

  13. Antiviral Activity of Graphene-Silver Nanocomposites against Non-Enveloped and Enveloped Viruses.

    PubMed

    Chen, Yi-Ning; Hsueh, Yi-Huang; Hsieh, Chien-Te; Tzou, Dong-Ying; Chang, Pai-Ling

    2016-04-01

    The discovery of novel antiviral materials is important because many infectious diseases are caused by viruses. Silver nanoparticles have demonstrated strong antiviral activity, and graphene is a potential antimicrobial material due to its large surface area, high carrier mobility, and biocompatibility. No studies on the antiviral activity of nanomaterials on non-enveloped viruses have been reported. To investigate the antiviral activity of graphene oxide (GO) sheets and GO sheets with silver particles (GO-Ag) against enveloped and non-enveloped viruses, feline coronavirus (FCoV) with an envelope and infectious bursal disease virus (IBDV) without an envelope were chosen. The morphology and sizes of GO and GO-Ag were characterized by transmission, scanning electron microscopy, and X-ray diffraction. A virus inhibition assay was used to identify the antiviral activity of GO and GO-Ag. Go-Ag inhibited 25% of infection by FCoV and 23% by IBDV, whereas GO only inhibited 16% of infection by FCoV but showed no antiviral activity against the infection by IBDV. Further application of GO and GO-Ag can be considered for personal protection equipment to decrease the transmission of viruses. PMID:27104546

  14. The Prolyl Isomerase Pin1 Promotes the Herpesvirus-Induced Phosphorylation-Dependent Disassembly of the Nuclear Lamina Required for Nucleocytoplasmic Egress

    PubMed Central

    Milbradt, Jens; Hutterer, Corina; Bahsi, Hanife; Wagner, Sabrina; Sonntag, Eric; Kaufer, Benedikt B.; Mori, Yasuko; Sticht, Heinrich; Fossen, Torgils; Marschall, Manfred

    2016-01-01

    The nuclear lamina lines the inner nuclear membrane providing a structural framework for the nucleus. Cellular processes, such as nuclear envelope breakdown during mitosis or nuclear export of large ribonucleoprotein complexes, are functionally linked to the disassembly of the nuclear lamina. In general, lamina disassembly is mediated by phosphorylation, but the precise molecular mechanism is still not completely understood. Recently, we suggested a novel mechanism for lamina disassembly during the nuclear egress of herpesviral capsids which involves the cellular isomerase Pin1. In this study, we focused on mechanistic details of herpesviral nuclear replication to demonstrate the general importance of Pin1 for lamina disassembly. In particular, Ser22-specific lamin phosphorylation consistently generates a Pin1-binding motif in cells infected with human and animal alpha-, beta-, and gammaherpesviruses. Using nuclear magnetic resonance spectroscopy, we showed that binding of Pin1 to a synthetic lamin peptide induces its cis/trans isomerization in vitro. A detailed bioinformatic evaluation strongly suggests that this structural conversion induces large-scale secondary structural changes in the lamin N-terminus. Thus, we concluded that a Pin1-induced conformational change of lamins may represent the molecular trigger responsible for lamina disassembly. Consistent with this concept, pharmacological inhibition of Pin1 activity blocked lamina disassembly in herpesvirus-infected fibroblasts and consequently impaired virus replication. In addition, a phospho-mimetic Ser22Glu lamin mutant was still able to form a regular lamina structure and overexpression of a Ser22-phosphorylating kinase did not induce lamina disassembly in Pin1 knockout cells. Intriguingly, this was observed in absence of herpesvirus infection proposing a broader importance of Pin1 for lamina constitution. Thus, our results suggest a functional model of similar events leading to disassembly of the nuclear

  15. The Prolyl Isomerase Pin1 Promotes the Herpesvirus-Induced Phosphorylation-Dependent Disassembly of the Nuclear Lamina Required for Nucleocytoplasmic Egress.

    PubMed

    Milbradt, Jens; Hutterer, Corina; Bahsi, Hanife; Wagner, Sabrina; Sonntag, Eric; Horn, Anselm H C; Kaufer, Benedikt B; Mori, Yasuko; Sticht, Heinrich; Fossen, Torgils; Marschall, Manfred

    2016-08-01

    The nuclear lamina lines the inner nuclear membrane providing a structural framework for the nucleus. Cellular processes, such as nuclear envelope breakdown during mitosis or nuclear export of large ribonucleoprotein complexes, are functionally linked to the disassembly of the nuclear lamina. In general, lamina disassembly is mediated by phosphorylation, but the precise molecular mechanism is still not completely understood. Recently, we suggested a novel mechanism for lamina disassembly during the nuclear egress of herpesviral capsids which involves the cellular isomerase Pin1. In this study, we focused on mechanistic details of herpesviral nuclear replication to demonstrate the general importance of Pin1 for lamina disassembly. In particular, Ser22-specific lamin phosphorylation consistently generates a Pin1-binding motif in cells infected with human and animal alpha-, beta-, and gammaherpesviruses. Using nuclear magnetic resonance spectroscopy, we showed that binding of Pin1 to a synthetic lamin peptide induces its cis/trans isomerization in vitro. A detailed bioinformatic evaluation strongly suggests that this structural conversion induces large-scale secondary structural changes in the lamin N-terminus. Thus, we concluded that a Pin1-induced conformational change of lamins may represent the molecular trigger responsible for lamina disassembly. Consistent with this concept, pharmacological inhibition of Pin1 activity blocked lamina disassembly in herpesvirus-infected fibroblasts and consequently impaired virus replication. In addition, a phospho-mimetic Ser22Glu lamin mutant was still able to form a regular lamina structure and overexpression of a Ser22-phosphorylating kinase did not induce lamina disassembly in Pin1 knockout cells. Intriguingly, this was observed in absence of herpesvirus infection proposing a broader importance of Pin1 for lamina constitution. Thus, our results suggest a functional model of similar events leading to disassembly of the nuclear

  16. The Prolyl Isomerase Pin1 Promotes the Herpesvirus-Induced Phosphorylation-Dependent Disassembly of the Nuclear Lamina Required for Nucleocytoplasmic Egress.

    PubMed

    Milbradt, Jens; Hutterer, Corina; Bahsi, Hanife; Wagner, Sabrina; Sonntag, Eric; Horn, Anselm H C; Kaufer, Benedikt B; Mori, Yasuko; Sticht, Heinrich; Fossen, Torgils; Marschall, Manfred

    2016-08-01

    The nuclear lamina lines the inner nuclear membrane providing a structural framework for the nucleus. Cellular processes, such as nuclear envelope breakdown during mitosis or nuclear export of large ribonucleoprotein complexes, are functionally linked to the disassembly of the nuclear lamina. In general, lamina disassembly is mediated by phosphorylation, but the precise molecular mechanism is still not completely understood. Recently, we suggested a novel mechanism for lamina disassembly during the nuclear egress of herpesviral capsids which involves the cellular isomerase Pin1. In this study, we focused on mechanistic details of herpesviral nuclear replication to demonstrate the general importance of Pin1 for lamina disassembly. In particular, Ser22-specific lamin phosphorylation consistently generates a Pin1-binding motif in cells infected with human and animal alpha-, beta-, and gammaherpesviruses. Using nuclear magnetic resonance spectroscopy, we showed that binding of Pin1 to a synthetic lamin peptide induces its cis/trans isomerization in vitro. A detailed bioinformatic evaluation strongly suggests that this structural conversion induces large-scale secondary structural changes in the lamin N-terminus. Thus, we concluded that a Pin1-induced conformational change of lamins may represent the molecular trigger responsible for lamina disassembly. Consistent with this concept, pharmacological inhibition of Pin1 activity blocked lamina disassembly in herpesvirus-infected fibroblasts and consequently impaired virus replication. In addition, a phospho-mimetic Ser22Glu lamin mutant was still able to form a regular lamina structure and overexpression of a Ser22-phosphorylating kinase did not induce lamina disassembly in Pin1 knockout cells. Intriguingly, this was observed in absence of herpesvirus infection proposing a broader importance of Pin1 for lamina constitution. Thus, our results suggest a functional model of similar events leading to disassembly of the nuclear

  17. Thermodynamics of nuclear transport

    NASA Astrophysics Data System (ADS)

    Wang, Ching-Hao; Mehta, Pankaj; Elbaum, Michael

    Molecular transport across the nuclear envelope is important for eukaryotes for gene expression and signaling. Experimental studies have revealed that nuclear transport is inherently a nonequilibrium process and actively consumes energy. In this work we present a thermodynamics theory of nuclear transport for a major class of nuclear transporters that are mediated by the small GTPase Ran. We identify the molecular elements responsible for powering nuclear transport, which we term the ``Ran battery'' and find that the efficiency of transport, measured by the cargo nuclear localization ratio, is limited by competition between cargo molecules and RanGTP to bind transport receptors, as well as the amount of NTF2 (i.e. RanGDP carrier) available to circulate the energy flow. This picture complements our current understanding of nuclear transport by providing a comprehensive thermodynamics framework to decipher the underlying biochemical machinery. Pm and CHW were supported by a Simons Investigator in the Mathematical Modeling in Living Systems grant (to PM).

  18. Three-dimensional visualization of the Autographa californica multiple nucleopolyhedrovirus occlusion-derived virion envelopment process gives new clues as to its mechanism

    SciTech Connect

    Shi, Yang; Li, Kunpeng; Tang, Peiping; Li, Yinyin; Zhou, Qiang; Yang, Kai; Zhang, Qinfen

    2015-02-15

    Baculoviruses produce two virion phenotypes, occlusion-derived virion (ODV) and budded virion (BV). ODV envelopment occurs in the nucleus. Morphogenesis of the ODV has been studied extensively; however, the mechanisms underlying microvesicle formation and ODV envelopment in nuclei remain unclear. In this study, we used electron tomography (ET) together with the conventional electron microscopy to study the envelopment of Autographa californica multiple nucleopolyhedrovirus (AcMNPV) ODV. Our results demonstrate that not only the inner but also the outer nuclear membrane can invaginate and vesiculate into microvesicles and that intranuclear microvesicles are the direct source of the ODV membrane. Five main events in the ODV envelopment process are summarized, from which we propose a model to explain this process. - Highlights: • Both the inner and outer nuclear membranes could invaginate. • Both the inner and outer nuclear membranes could vesiculate into microvesicles. • Five main events in the ODV envelopment process are summarized. • A model is proposed to explain this ODV envelopment.

  19. Analysis Code for High Gradient Dielectric Insulator Surface Breakdown

    SciTech Connect

    Ives, Robert Lawrence; Verboncoeur, John; Aldan, Manuel

    2010-05-30

    High voltage (HV) insulators are critical components in high-energy, accelerator and pulsed power systems that drive diverse applications in the national security, nuclear weapons science, defense and industrial arenas. In these systems, the insulator may separate vacuum/non-vacuum regions or conductors with high electrical field gradients. These insulators will often fail at electric fields over an order of magnitude lower than their intrinsic dielectric strength due to flashover at the dielectric interface. Decades of studies have produced a wealth of information on fundamental processes and mechanisms important for flashover initiation, but only for relatively simple insulator configurations in controlled environments. Accelerator and pulsed power system designers are faced with applying the fundamental knowledge to complex, operational devices with escalating HV requirements. Designers are forced to rely on “best practices” and expensive prototype testing, providing boundaries for successful operation. However, the safety margin is difficult to estimate, and system design must be very conservative for situations where testing is not practicable, or replacement of failed parts is disruptive or expensive. The Phase I program demonstrated the feasibility of developing an advanced code for modeling insulator breakdown. Such a code would be of great interest for a number of applications, including high energy physics, microwave source development, fusion sciences, and other research and industrial applications using high voltage devices.

  20. Confusion and its dynamics during device comprehension with breakdown scenarios.

    PubMed

    D'Mello, Sidney; Graesser, Art

    2014-09-01

    The incidence and dynamics of confusion during complex learning and problem solving were investigated in an experiment where participants first read illustrated texts on everyday devices (e.g., an electric bell) followed by breakdown scenarios reflecting device malfunctions (e.g., "When a person rang the bell there was a short ding and then no sound was heard"). The breakdown scenarios were expected to trigger impasses and put participants in a state of cognitive disequilibrium where they would experience confusion and engage in effortful confusion resolution activities in order to restore equilibrium. The results confirmed that participants reported more confusion when presented with the breakdown scenarios compared to control scenarios that involved focusing on important device components in the absence of malfunctions. A second-by-second analysis of the dynamics of confusion yielded two characteristic trajectories that distinguished participants who partially resolved their confusion from those who remained confused. Participants who were successful in partial confusion resolution while processing the breakdowns outperformed their counterparts on knowledge assessments after controlling for scholastic aptitude, engagement, and frustration. This effect was amplified for those who were highly confused by the breakdowns. There was no direct breakdown vs. control effect on learning, but being actively engaged and partially resolving confusion during breakdown processing were positive predictors of increased learning with the breakdown compared to control scenarios. Implications of our findings for theories that highlight the role of impasses, cognitive disequilibrium, and confusion to learning are discussed.

  1. Experimental Study on Electrical Breakdown for Devices with Micrometer Gaps

    NASA Astrophysics Data System (ADS)

    Meng, Guodong; Cheng, Yonghong; Dong, Chengye; Wu, Kai

    2014-12-01

    The understanding of electrical breakdown in atmospheric air across micrometer gaps is critically important for the insulation design of micro & nano electronic devices. In this paper, planar aluminum electrodes with gaps ranging from 2 μm to 40 μm were fabricated by microelectromechanical system technology. The influence factors including gap width and surface dielectric states were experimentally investigated using the home-built test and measurement system. Results showed that for SiO2 layers the current sustained at 2-3 nA during most of the pre-breakdown period, and then rose rapidly to 10-30 nA just before breakdown due to field electron emission, followed by the breakdown. The breakdown voltage curves demonstrated three stages: (1) a constantly decreasing region (the gap width d < 5 μm), where the field emission effect played an important role just near breakdown, supplying enough initial electrons for the breakdown process; (2) a plateau region with a near constant breakdown potential (5 μm < d < 10 μm) (3) a region for large gaps that adhered to Paschen's curve (d > 10 μm). And the surface dielectric states including the surface resistivity and secondary electron yield were verified to be related to the propagation of discharge due to the interaction between initial electrons and dielectrics.

  2. Modeling optical breakdown in dielectrics during ultrafast laser processing.

    PubMed

    Fan, C H; Longtin, J P

    2001-06-20

    Laser ablation is widely used in micromachining, manufacturing, thin-film formation, and bioengineering applications. During laser ablation the removal of material and quality of the features depend strongly on the optical breakdown region induced by the laser irradiance. The recent advent of amplified ultrafast lasers with pulse durations of less than 1 ps has generated considerable interest because of the ability of the lasers to process virtually all materials with high precision and minimal thermal damage. With ultrashort pulse widths, however, traditional breakdown models no longer accurately capture the laser-material interaction that leads to breakdown. A femtosecond breakdown model for dielectric solids and liquids is presented that characterizes the pulse behavior and predicts the time- and position-dependent breakdown region. The model includes the pulse propagation and small spatial extent of ultrashort laser pulses. Model results are presented and compared with classical breakdown models for 1-ns, 1-ps, and 150-fs pulses. The results show that the revised model is able to model breakdown accurately in the focal region for pulse durations of less than 10 ps. The model can also be of use in estimating the time- and position-resolved electron density in the interaction volume, the breakdown threshold of the material, shielding effects, and temperature distributions during ultrafast processing. PMID:18357333

  3. 7 CFR 51.1009 - Stylar end breakdown.

    Code of Federal Regulations, 2011 CFR

    2011-01-01

    ... 7 Agriculture 2 2011-01-01 2011-01-01 false Stylar end breakdown. 51.1009 Section 51.1009... STANDARDS) United States Standards for Persian (Tahiti) Limes Definitions § 51.1009 Stylar end breakdown... affected area becomes darker and usually sinks below the healthy surface, but the area remains firm...

  4. 19 CFR 141.87 - Breakdown on component materials.

    Code of Federal Regulations, 2012 CFR

    2012-04-01

    ... 19 Customs Duties 2 2012-04-01 2012-04-01 false Breakdown on component materials. 141.87 Section 141.87 Customs Duties U.S. CUSTOMS AND BORDER PROTECTION, DEPARTMENT OF HOMELAND SECURITY; DEPARTMENT OF THE TREASURY (CONTINUED) ENTRY OF MERCHANDISE Invoices § 141.87 Breakdown on component...

  5. 7 CFR 51.1009 - Stylar end breakdown.

    Code of Federal Regulations, 2010 CFR

    2010-01-01

    ... 7 Agriculture 2 2010-01-01 2010-01-01 false Stylar end breakdown. 51.1009 Section 51.1009... STANDARDS) United States Standards for Persian (Tahiti) Limes Definitions § 51.1009 Stylar end breakdown... affected area becomes darker and usually sinks below the healthy surface, but the area remains firm...

  6. Vortex breakdown in a truncated conical bioreactor

    NASA Astrophysics Data System (ADS)

    Balci, Adnan; Brøns, Morten; Herrada, Miguel A.; Shtern, Vladimir N.

    2015-12-01

    This numerical study explains the eddy formation and disappearance in a slow steady axisymmetric air-water flow in a vertical truncated conical container, driven by the rotating top disk. Numerous topological metamorphoses occur as the water height, Hw, and the bottom-sidewall angle, α, vary. It is found that the sidewall convergence (divergence) from the top to the bottom stimulates (suppresses) the development of vortex breakdown (VB) in both water and air. At α = 60°, the flow topology changes eighteen times as Hw varies. The changes are due to (a) competing effects of AMF (the air meridional flow) and swirl, which drive meridional motions of opposite directions in water, and (b) feedback of water flow on AMF. For small Hw, the AMF effect dominates. As Hw increases, the swirl effect dominates and causes VB. The water flow feedback produces and modifies air eddies. The results are of fundamental interest and can be relevant for aerial bioreactors.

  7. Laser-induced breakdown spectroscopy in China

    NASA Astrophysics Data System (ADS)

    Wang, Zhe; Yuan, Ting-Bi; Hou, Zong-Yu; Zhou, Wei-Dong; Lu, Ji-Dong; Ding, Hong-Bin; Zeng, Xiao-Yan

    2014-08-01

    Laser-induced breakdown spectroscopy (LIBS) has been regarded as a future superstar for chemical analysis for years due to its unique features such as little or no sample preparation, remote sensing, and fast and multi-element analysis. Chinese LIBS community is one of the most dynamically developing communities in the World. The aim of the work is to inspect what have been done in China for LIBS development and, based on the understanding of the overall status, to identify the challenges and opportunities for the future development. In this paper, the scientific contributions from Chinese LIBS community are reviewed for the following four aspects: fundamentals, instrumentation, data processing and modeling, and applications; and the driving force of LIBS development in China is analyzed, the critical issues for successful LIBS application are discussed, and in our opinion, the potential direction to improve the technology and to realize large scale commercialization in China is proposed.

  8. State Regulation, Family Breakdown, and Lone Motherhood

    PubMed Central

    Meek, Jeff

    2014-01-01

    Using a range of parish records, records from the Registrar General of Scotland, charity organizations, and media reports, this article contributes to the historiography which evaluates the effects of World War I in Britain as well as the history of lone mothers and their children. It highlights how during the war, women, especially lone mothers, made significant gains through the welfare system, changing approaches to illegitimacy and the plentiful nature of women’s work but also how in doing so this brought them under greater surveillance by the state, local parishes, and charity organizations. Moreover, as this article will demonstrate, many of the gains made by women were short-lived and in fact the war contributed to high levels of family breakdown and gendered and intergenerational poverty endured by lone mothers and their children. PMID:26538794

  9. Runaway breakdown and hydrometeors in lightning initiation.

    PubMed

    Gurevich, A V; Karashtin, A N

    2013-05-01

    The particular electric pulse discharges are observed in thunderclouds during the initiation stage of negative cloud-to-ground lightning. The discharges are quite different from conventional streamers or leaders. A detailed analysis reveals that the shape of the pulses is determined by the runaway breakdown of air in the thundercloud electric field initiated by extensive atmospheric showers (RB-EAS). The high amplitude of the pulse electric current is due to the multiple microdischarges at hydrometeors stimulated and synchronized by the low-energy electrons generated in the RB-EAS process. The series of specific pulse discharges leads to charge reset from hydrometeors to the free ions and creates numerous stretched ion clusters, both positive and negative. As a result, a wide region in the thundercloud with a sufficiently high fractal ion conductivity is formed. The charge transport by ions plays a decisive role in the lightning leader preconditioning. PMID:23683210

  10. Vortex Breakdown-Aircraft Tail Interaction

    NASA Astrophysics Data System (ADS)

    Kim, Younjong; Rockwell, Donald

    2003-11-01

    The interaction of vortex breakdown with the tail of an aircraft can lead to severe unsteady loading and vibration. A technique of high-image-density particle image velocimetry is employed to characterize the instantaneous and averaged structure of a broken-down vortex with a generic tail configuration. Interaction of the primary (incident) vortex with the tail results in formation of a relatively large-scale cluster of secondary vorticity. The coexistence of these primary and secondary vortical structures is intimately associated with the unsteadiness of the vortex system, and thereby the near-surface fluctuations associated with buffet loading. Instantaneous and averaged representations of the vortex-tail interaction provide insight into the complex physics. Furthermore, a low order POD model is employed to characterize the most energetic modes of the vortex-tail interaction.

  11. Turbulent plumes in stellar convective envelopes.

    NASA Astrophysics Data System (ADS)

    Rieutord, M.; Zahn, J.-P.

    1995-04-01

    Recent numerical simulations of compressible convection in a stratified medium suggest that strong downwards directed flows may play an important role in stellar convective envelopes, both in the dynamics and in the energy transport. We transpose this idea to stellar convective envelopes by assuming that these plumes are turbulent plumes which may be described by Taylor's entrainment hypothesis, whose validity is well established in various geophysical conditions. We consider first the ideal case of turbulent plumes occurring in an isentropic atmosphere, and ignore all types of feedback. Thereafter we include the effect of the backflow generated by the plumes, and take into account the contribution of the radiative flux. The main result is that plumes originating from the upper layers of a star are able to reach the base of its convective envelope. Their number is necessarily limited because of their conical shape; the backflow further reduces their number to a maximum of about 1000. In these plumes the flux of kinetic energy is directed downwards, but it is less than the upwards directed enthalpy flux, so that the plumes always carry a net energy flux towards the surface. Our plume model is not applicable near the surface, where the departures from adiabaticity become important due to radiative leaking; therefore it cannot predict the depth of the convection zone, which is determined mainly by the transition from the radiative regime above to the nearly adiabatic conditions below. Neither does it permit to evaluate the extent of penetration, which strongly depends on the (unknown) number of plumes. We conclude that, to be complete, a phenomenological model of stellar convection must have a dual character: it should include both the advective transport through diving plumes, which is outlined in this paper, and the turbulent diffusion achieved by the interstitial medium. Only the latter process is apprehended by the familiar mixing-length treatment.

  12. Quasiparticle breakdown in a quantum spin liquid.

    PubMed

    Stone, Matthew B; Zaliznyak, Igor A; Hong, Tao; Broholm, Collin L; Reich, Daniel H

    2006-03-01

    Much of modern condensed matter physics is understood in terms of elementary excitations, or quasiparticles--fundamental quanta of energy and momentum. Various strongly interacting atomic systems are successfully treated as a collection of quasiparticles with weak or no interactions. However, there are interesting limitations to this description: in some systems the very existence of quasiparticles cannot be taken for granted. Like unstable elementary particles, quasiparticles cannot survive beyond a threshold where certain decay channels become allowed by conservation laws; their spectrum terminates at this threshold. Such quasiparticle breakdown was first predicted for an exotic state of matter--super-fluid 4He at temperatures close to absolute zero, a quantum Bose liquid where zero-point atomic motion precludes crystallization. Here we show, using neutron scattering, that quasiparticle breakdown can also occur in a quantum magnet and, by implication, in other systems with Bose quasiparticles. We have measured spin excitations in a two-dimensional quantum magnet, piperazinium hexachlorodicuprate (PHCC), in which spin-1/2 copper ions form a non-magnetic quantum spin liquid, and find remarkable similarities with excitations in superfluid 4He. We observe a threshold momentum beyond which the quasiparticle peak merges with the two-quasiparticle continuum. It then acquires a finite energy width and becomes indistinguishable from a leading-edge singularity, so that excited states are no longer quasiparticles but occupy a wide band of energy. Our findings have important ramifications for understanding excitations with gapped spectra in many condensed matter systems, ranging from band insulators to high-transition-temperature superconductors.

  13. Quasiparticle breakdown in a quantum spin liquid.

    PubMed

    Stone, Matthew B; Zaliznyak, Igor A; Hong, Tao; Broholm, Collin L; Reich, Daniel H

    2006-03-01

    Much of modern condensed matter physics is understood in terms of elementary excitations, or quasiparticles--fundamental quanta of energy and momentum. Various strongly interacting atomic systems are successfully treated as a collection of quasiparticles with weak or no interactions. However, there are interesting limitations to this description: in some systems the very existence of quasiparticles cannot be taken for granted. Like unstable elementary particles, quasiparticles cannot survive beyond a threshold where certain decay channels become allowed by conservation laws; their spectrum terminates at this threshold. Such quasiparticle breakdown was first predicted for an exotic state of matter--super-fluid 4He at temperatures close to absolute zero, a quantum Bose liquid where zero-point atomic motion precludes crystallization. Here we show, using neutron scattering, that quasiparticle breakdown can also occur in a quantum magnet and, by implication, in other systems with Bose quasiparticles. We have measured spin excitations in a two-dimensional quantum magnet, piperazinium hexachlorodicuprate (PHCC), in which spin-1/2 copper ions form a non-magnetic quantum spin liquid, and find remarkable similarities with excitations in superfluid 4He. We observe a threshold momentum beyond which the quasiparticle peak merges with the two-quasiparticle continuum. It then acquires a finite energy width and becomes indistinguishable from a leading-edge singularity, so that excited states are no longer quasiparticles but occupy a wide band of energy. Our findings have important ramifications for understanding excitations with gapped spectra in many condensed matter systems, ranging from band insulators to high-transition-temperature superconductors. PMID:16525467

  14. Rotation prevents finite-time breakdown

    NASA Astrophysics Data System (ADS)

    Liu, Hailiang; Tadmor, Eitan

    2004-02-01

    We consider a two-dimensional (2D) convection model augmented with the rotational Coriolis forcing, Ut+ U·∇ xU=2 kU⊥, with a fixed 2 k being the inverse Rossby number. We ask whether the action of dispersive rotational forcing alone, U⊥, prevents the generic finite-time breakdown of the free nonlinear convection. The answer provided in this work is a conditional yes. Namely, we show that the rotating Euler equations admit global smooth solutions for a subset of generic initial configurations. With other configurations, however, finite-time breakdown of solutions may and actually does occur. Thus, global regularity depends on whether the initial configuration crosses an intrinsic, O(1) critical threshold (CT), which is quantified in terms of the initial vorticity, ω0=∇× U0, and the initial spectral gap associated with the 2×2 initial velocity gradient, η0≔ λ2(0)- λ1(0), λj(0)= λj(∇ U0). Specifically, global regularity of the rotational Euler equation is ensured if and only if 4kω 0(α)+η 20(α)<4k 2,∀α∈ R2. We also prove that the velocity field remains smooth if and only if it is periodic. An equivalent Lagrangian formulation reconfirms the CT and shows a global periodicity of velocity field as well as the associated particle orbits. Moreover, we observe yet another remarkable periodic behavior exhibited by the gradient of the velocity field. The spectral dynamics of the Eulerian formulation [SIAM J. Math. Anal. 33 (2001) 930] reveals that the vorticity and the divergence of the flow evolve with their own path-dependent period. We conclude with a kinetic formulation of the rotating Euler equation.

  15. Surface area coefficients for airship envelopes

    NASA Technical Reports Server (NTRS)

    Diehl, W S

    1922-01-01

    In naval architecture, it is customary to determine the wetted surface of a ship by means of some formula which involves the principal dimensions of the design and suitable constants. These formulas of naval architecture may be extended and applied to the calculation of the surface area of airship envelopes by the use of new values of the constants determined for this purpose. Surface area coefficients were calculated from the actual dimensions, surfaces, and volumes of 52 streamline bodies, which form a series covering the entire range of shapes used in the present aeronautical practice.

  16. Low heat-leak cryogenic envelope

    DOEpatents

    DeHaan, James R.

    1976-10-19

    A plurality of cryogenic envelope sections are joined together to form a power transmission line. Each of the sections is comprised of inner and outer tubes having multilayer metalized plastic spirally wrapped within a vacuum chamber formed between the inner and outer tubes. A refrigeration tube traverses the vacuum chamber, but exits one section and enters another through thermal standoffs for reducing heat-leak from the outer tube to the refrigeration tube. The refrigeration tube passes through a spirally wrapped shield within each section's vacuum chamber in a manner so that the refrigeration tube is in close thermal contact with the shield, but is nevertheless slideable with respect thereto.

  17. Snell Envelope with Small Probability Criteria

    SciTech Connect

    Del Moral, Pierre Hu, Peng; Oudjane, Nadia

    2012-12-15

    We present a new algorithm to compute the Snell envelope in the specific case where the criteria to optimize is associated with a small probability or a rare event. This new approach combines the Stochastic Mesh approach of Broadie and Glasserman with a particle approximation scheme based on a specific change of measure designed to concentrate the computational effort in regions pointed out by the criteria. The theoretical analysis of this new algorithm provides non asymptotic convergence estimates. Finally, the numerical tests confirm the practical interest of this approach.

  18. Detection and modeling of rough component envelopes

    NASA Astrophysics Data System (ADS)

    Peacock, Matthew; Dluzniak, Richard; Thompson, William

    1995-08-01

    This paper describes an imaging technique for the determination of rough component envelopes of cast and forged components. The paper includes several image acquisition methods currently used in this area but concentrates in detail on the method known as the light stripe method. Results presented show the advantages of the light stripe method to obtain a fast and accurate 3D description of the cast and forged components. The research is part of a larger project on intelligent manufacturing systems and is being conducted at the CIM Centre, Swinburne University of Technology, Hawthorn, Victoria, Australia.

  19. On the electric breakdown in liquid argon at centimeter scale

    NASA Astrophysics Data System (ADS)

    Auger, M.; Blatter, A.; Ereditato, A.; Goeldi, D.; Janos, S.; Kreslo, I.; Luethi, M.; von Rohr, C. Rudolf; Strauss, T.; Weber, M. S.

    2016-03-01

    We present a study on the dependence of electric breakdown discharge properties on electrode geometry and the breakdown field in liquid argon near its boiling point. The measurements were performed with a spherical cathode and a planar anode at distances ranging from 0.1 mm to 10.0 mm. A detailed study of the time evolution of the breakdown volt-ampere characteristics was performed for the first time. It revealed a slow streamer development phase in the discharge. The results of a spectroscopic study of the visible light emission of the breakdowns complement the measurements. The light emission from the initial phase of the discharge is attributed to electro-luminescence of liquid argon following a current of drifting electrons. These results contribute to set benchmarks for breakdown-safe design of ionization detectors, such as Liquid Argon Time Projection Chambers (LAr TPC).

  20. Breakdown voltage of metal-oxide resistors in liquid argon

    SciTech Connect

    Bagby, L. F.; Gollapinni, S.; James, C. C.; Jones, B. J.P.; Jostlein, H.; Lockwitz, S.; Naples, D.; Raaf, J. L.; Rameika, R.; Schukraft, A.; Strauss, T.; Weber, M. S.; Wolbers, S. A.

    2014-11-07

    We characterized a sample of metal-oxide resistors and measured their breakdown voltage in liquid argon by applying high voltage (HV) pulses over a 3 second period. This test mimics the situation in a HV-divider chain when a breakdown occurs and the voltage across resistors rapidly rise from the static value to much higher values. All resistors had higher breakdown voltages in liquid argon than their vendor ratings in air at room temperature. Failure modes range from full destruction to coating damage. In cases where breakdown was not catastrophic, subsequent breakdown voltages were lower in subsequent measuring runs. One resistor type withstands 131 kV pulses, the limit of the test setup.

  1. Development of Hybrid Product Breakdown Structure for NASA Ground Systems

    NASA Technical Reports Server (NTRS)

    Monaghan, Mark W.; Henry, Robert J.

    2013-01-01

    The Product Breakdown Structure is traditionally a method of identification of the products of a project in a tree structure. It is a tool used to assess, plan, document, and display the equipment requirements for a project. It is part of a product based planning technique, and attempts to break down all components of a project in as much detail as possible, so that nothing is overlooked. The PBS for ground systems at the Kennedy Space Center is being developed to encompass the traditional requirements including the alignment of facility, systems, and components to the organizational hierarchy. The Ground Operations Product Breakdown Structure is a hybrid in nature in that some aspects of a work breakdown structure will be incorporated and merged with the Architecture Concept of Operations, Master Subsystem List, customer interface, and assigned management responsibility. The Ground Operations Product Breakdown Structure needs to be able to identify the flexibility of support differing customers (internal and external) usage of ground support equipment within the Kennedy Space Center launch and processing complex. The development of the Product Breakdown Structure is an iterative activity Initially documenting the organization hierarchy structure and relationships. The Product Breakdown Structure identifies the linkage between the customer program requirements, allocation of system resources, development of design goals, and identification logistics products. As the Product Breakdown Structure progresses the incorporation of the results of requirement planning for the customer occurs identifying facility needs and systems. The mature Product Breakdown Structure is baselined with a hierarchical drawing, the Product Breakdown Structure database, and an associated document identifying the verification of the data through the life cycle of the program/product line. This paper will document, demonstrate, and identify key aspects of the life cycle of a Hybrid Product

  2. Nuclear disintegration in relativistic heavy-ion collisions

    SciTech Connect

    Gutbrod, H.H.; Warwick, A.I.; Wieman, H.

    1982-04-01

    The breakdown of the participant spectator model for central relativistic nuclear collisions is discussed and a different picture of a hot spot followed by a target explosion is suggested to be more consistent with the data.

  3. Effects of lyophilization on the infectivity of enveloped and non-enveloped viruses in bone tissue.

    PubMed

    Uhlenhaut, Christine; Dörner, Thomas; Pauli, Georg; Pruss, Axel

    2005-11-01

    Recently reported qualitative experiments proved that retroviral infectivity is not destroyed by lyophilization performed on systemically infected bone and tendon. The now accomplished quantitative determination of residual infectivity for enveloped and non-enveloped viruses allows a validation of the production process regarding viral safety in freeze-dried bone transplants. The lyophilization effect on the infectivity of two non-enveloped viruses (Maus Elberfeld virus, MEV; Porcine parvovirus, PPV) and one enveloped virus (Vesicular Stomatitis virus, VSV) was examined for virus-spiked bone material in comparison to lyophilized viruses, original virus stock, and air-dried viruses. All experiments were carried out with both cell-free and cell-associated virus. Significant differences were observed regarding the reduction of virus titers (TCID50). Infectivity of VSV was reduced by about 3-4 log10 using lyophilization in presence of bone matrix and of MEV by 6-7 log10, while no substantial reduction in virus titers was observed for PPV. Lyophilization of cell-free or cell-associated virus is not sufficient to inactivate viruses completely. However, lyophilization could have an additive effect in line with other production steps used in the manufacturing process.

  4. Envelope as Climate Negotiator: Evaluating adaptive building envelope's capacity to moderate indoor climate and energy

    NASA Astrophysics Data System (ADS)

    Erickson, James

    Through manipulation of adaptable opportunities available within a given environment, individuals become active participants in managing personal comfort requirements, by exercising control over their comfort without the assistance of mechanical heating and cooling systems. Similarly, continuous manipulation of a building skin's form, insulation, porosity, and transmissivity qualities exerts control over the energy exchanged between indoor and outdoor environments. This research uses four adaptive response variables in a modified software algorithm to explore an adaptive building skin's potential in reacting to environmental stimuli with the purpose of minimizing energy use without sacrificing occupant comfort. Results illustrate that significant energy savings can be realized with adaptive envelopes over static building envelopes even under extreme summer and winter climate conditions; that the magnitude of these savings are dependent on climate and orientation; and that occupant thermal comfort can be improved consistently over comfort levels achieved by optimized static building envelopes. The resulting adaptive envelope's unique climate-specific behavior could inform designers in creating an intelligent kinetic aesthetic that helps facilitate adaptability and resiliency in architecture.

  5. On-Line Safe Flight Envelope Determination for Impaired Aircraft

    NASA Technical Reports Server (NTRS)

    Lombaerts, Thomas; Schuet, Stefan; Acosta, Diana; Kaneshige, John

    2015-01-01

    The design and simulation of an on-line algorithm which estimates the safe maneuvering envelope of aircraft is discussed in this paper. The trim envelope is estimated using probabilistic methods and efficient high-fidelity model based computations of attainable equilibrium sets. From this trim envelope, a robust reachability analysis provides the maneuverability limitations of the aircraft through an optimal control formulation. Both envelope limits are presented to the flight crew on the primary flight display. In the results section, scenarios are considered where this adaptive algorithm is capable of computing online changes to the maneuvering envelope due to impairment. Furthermore, corresponding updates to display features on the primary flight display are provided to potentially inform the flight crew of safety critical envelope alterations caused by the impairment.

  6. Groupwise Dimension Reduction via Envelope Method

    PubMed Central

    Guo, Zifang; Li, Lexin; Lu, Wenbin; Li, Bing

    2016-01-01

    The family of sufficient dimension reduction (SDR) methods that produce informative combinations of predictors, or indices, are particularly useful for high dimensional regression analysis. In many such analyses, it becomes increasingly common that there is available a priori subject knowledge of the predictors; e.g., they belong to different groups. While many recent SDR proposals have greatly expanded the scope of the methods’ applicability, how to effectively incorporate the prior predictor structure information remains a challenge. In this article, we aim at dimension reduction that recovers full regression information while preserving the predictor group structure. Built upon a new concept of the direct sum envelope, we introduce a systematic way to incorporate the group information in most existing SDR estimators. As a result, the reduction outcomes are much easier to interpret. Moreover, the envelope method provides a principled way to build a variety of prior structures into dimension reduction analysis. Both simulations and real data analysis demonstrate the competent numerical performance of the new method. PMID:26973362

  7. Precision envelope detector and linear rectifier circuitry

    DOEpatents

    Davis, Thomas J.

    1980-01-01

    Disclosed is a method and apparatus for the precise linear rectification and envelope detection of oscillatory signals. The signal is applied to a voltage-to-current converter which supplies current to a constant current sink. The connection between the converter and the sink is also applied through a diode and an output load resistor to a ground connection. The connection is also connected to ground through a second diode of opposite polarity from the diode in series with the load resistor. Very small amplitude voltage signals applied to the converter will cause a small change in the output current of the converter, and the difference between the output current and the constant current sink will be applied either directly to ground through the single diode, or across the output load resistor, dependent upon the polarity. Disclosed also is a full-wave rectifier utilizing constant current sinks and voltage-to-current converters. Additionally, disclosed is a combination of the voltage-to-current converters with differential integrated circuit preamplifiers to boost the initial signal amplitude, and with low pass filtering applied so as to obtain a video or signal envelope output.

  8. Functional organization of the HIV lipid envelope

    PubMed Central

    Huarte, Nerea; Carravilla, Pablo; Cruz, Antonio; Lorizate, Maier; Nieto-Garai, Jon A.; Kräusslich, Hans-Georg; Pérez-Gil, Jesús; Requejo-Isidro, Jose; Nieva, José L.

    2016-01-01

    The chemical composition of the human immunodeficiency virus type 1 (HIV-1) membrane is critical for fusion and entry into target cells, suggesting that preservation of a functional lipid bilayer organization may be required for efficient infection. HIV-1 acquires its envelope from the host cell plasma membrane at sites enriched in raft-type lipids. Furthermore, infectious particles display aminophospholipids on their surface, indicative of dissipation of the inter-leaflet lipid asymmetry metabolically generated at cellular membranes. By combining two-photon excited Laurdan fluorescence imaging and atomic force microscopy, we have obtained unprecedented insights into the phase state of membranes reconstituted from viral lipids (i.e., extracted from infectious HIV-1 particles), established the role played by the different specimens in the mixtures, and characterized the effects of membrane-active virucidal agents on membrane organization. In determining the molecular basis underlying lipid packing and lateral heterogeneity of the HIV-1 membrane, our results may help develop compounds with antiviral activity acting by perturbing the functional organization of the lipid envelope. PMID:27678107

  9. Transparent Helium in Stripped Envelope Supernovae

    NASA Astrophysics Data System (ADS)

    Piro, Anthony L.; Morozova, Viktoriya S.

    2014-09-01

    Using simple arguments based on photometric light curves and velocity evolution, we propose that some stripped envelope supernovae (SNe) show signs that a significant fraction of their helium is effectively transparent. The main pieces of evidence are the relatively low velocities with little velocity evolution, as are expected deep inside an exploding star, along with temperatures that are too low to ionize helium. This means that the helium should not contribute to the shaping of the main SN light curve, and thus the total helium mass may be difficult to measure from simple light curve modeling. Conversely, such modeling may be more useful for constraining the mass of the carbon/oxygen core of the SN progenitor. Other stripped envelope SNe show higher velocities and larger velocity gradients, which require an additional opacity source (perhaps the mixing of heavier elements or radioactive nickel) to prevent the helium from being transparent. We discuss ways in which similar analysis can provide insights into the differences and similarities between SNe Ib and Ic, which will lead to a better understanding of their respective formation mechanisms.

  10. TRANSPARENT HELIUM IN STRIPPED ENVELOPE SUPERNOVAE

    SciTech Connect

    Piro, Anthony L.; Morozova, Viktoriya S.

    2014-09-01

    Using simple arguments based on photometric light curves and velocity evolution, we propose that some stripped envelope supernovae (SNe) show signs that a significant fraction of their helium is effectively transparent. The main pieces of evidence are the relatively low velocities with little velocity evolution, as are expected deep inside an exploding star, along with temperatures that are too low to ionize helium. This means that the helium should not contribute to the shaping of the main SN light curve, and thus the total helium mass may be difficult to measure from simple light curve modeling. Conversely, such modeling may be more useful for constraining the mass of the carbon/oxygen core of the SN progenitor. Other stripped envelope SNe show higher velocities and larger velocity gradients, which require an additional opacity source (perhaps the mixing of heavier elements or radioactive nickel) to prevent the helium from being transparent. We discuss ways in which similar analysis can provide insights into the differences and similarities between SNe Ib and Ic, which will lead to a better understanding of their respective formation mechanisms.

  11. Solution of K-V envelope equations

    SciTech Connect

    Anderson, O.A.

    1995-04-01

    The envelope equations for a KV beam with space charge have been analyzed systematically by an e expansion followed by integrations. The focusing profile as a function of axial length is assumed to be symmetric but otherwise arbitrary. Given the bean current, emittance, and peak focusing field, we find the envelopes a(s) and b(s) and obtain , a{sub max}, {sigma}, and {sigma}{sub 0}. Explicit results are presented for various truncations of the expansion. The zeroth order results correspond to those from the well-known smooth approximation; the same convenient format is retained for the higher order cases. The first order results, involving single correction terms, give 3--10 times better accuracy and are good to {approximately}1% at {sigma}{sub 0} = 70{degree}. Third order gives a factor of 10--30 improvement over the smooth approximation and derived quantities accurate to {approximately}1% at {sigma}{sub 0} = 112 {degree}. The first order expressions are convenient design tools. They lend themselves to variable energy problems and have been applied to the design, construction, and testing of ESQ accelerators at LBL.

  12. From Circumstellar Envelopes to the Interstellar Medium

    NASA Astrophysics Data System (ADS)

    Ueta, T.

    2011-09-01

    IRAS and ISO observations have indicated that the extended dust shells of AGB stars could be explained by a simple model of a constant past mass outflow piling up at the interface with the interstellar medium (ISM). Recent Spitzer observations have shown that even outflows from AGB stars can induce shocks at the ISM-AGB wind interface, while a recent AKARI survey of the circumstellar envelopes of evolved stars have revealed far-IR structures resembling to the interface regions between the ISM and AGB winds in many objects. There have been observations made in other wavelengths (especially in the UV with GALEX) that corroborate these findings in the far-IR. New Herschel observations are expected to provide detailed views of these interface regions. Therefore, the extended dust shells of AGB stars should not only allow us to prove the mass-loss history of the parent AGB stars but also permit us to glimpse how the ejecta eventually merge with the ISM. In this review, I will summarize recent research developments made by observations with Spitzer, AKARI, and Herschel at the interface region between the circumstellar envelopes of AGB stars and the ISM.

  13. Glycolate transporter of the pea chloroplast envelope

    SciTech Connect

    Howitz, K.T.

    1985-01-01

    The discovery of a glycolate transporter in the pea (Pisum sativum) chloroplast envelope is described. Several novel silicone oil centrifugation methods were developed to resolve the initial rate kinetics of (/sup 14/C)glycolate transport by isolated, intact pea chloroplasts. Chloroplast glycolate transport was found to be carrier mediated. Transport rates saturated with increasing glycolate concentration. N-Ethylmaleimide (NEM) pretreatment of chloroplasts inhibited transport, an inhibition prevented by glycolate. Glycolate distributed across the envelope in a way which equalized stromal and medium glycolic acid concentrations, limiting possible transport mechanisms to facilitated glycolic acid diffusion, proton symport or hydroxyl antiport. The effects of stomal and medium pH's on the K/sub m/ and V/sub max/ fit the predictions of mobile carrier kinetic models of hydroxyl antiport or proton symport (H/sup +/ binds first). The carrier mediated transport was fast enough to be consistent with in vivo rates of photorespiration. The 2-hydroxymonocarboxylates, glycerate, lactate and glyoxylate are competitive inhibitors of chloroplast glycolate uptake. Glyoxylate, D-lactate and D-glycerate cause glycolate counterflow, indicating that they are also substrates of the glycolate carrier. This finding was confirmed for D-glycerate by studies on glycolate effects on (1-/sup 14/C)D-glycerate transport.

  14. Sensitivity to changes in amplitude envelope

    NASA Astrophysics Data System (ADS)

    Gallun, Erick; Hafter, Ervin R.; Bonnel, Anne-Marie

    2002-05-01

    Detection of a brief increment in a tonal pedestal is less well predicted by energy-detection (e.g., Macmillan, 1973; Bonnel and Hafter, 1997) than by sensitivity to changes in the stimulus envelope. As this implies a mechanism similar to an envelope extractor (Viemeister, 1979), sinusoidal amplitude modulation was used to mask a single ramped increment (10, 45, or 70 ms) added to a 1000-ms pedestal with carrier frequency (cf)=477 Hz. As in informational masking (Neff, 1994) and ``modulation-detection interference'' (Yost and Sheft, 1989), interference occurred with masker cfs of 477 and 2013 Hz. While slight masking was found with modulation frequencies (mfs) from 16 to 96 Hz, masking grew inversely with still lower mfs, being greatest for mf=4 Hz. This division is reminiscent of that said to separate sensations of ``roughness'' and ``beats,'' respectively (Terhardt, 1974), with the latter also being related to durations associated with auditory groupings in music and speech. Importantly, this result held for all of the signal durations and onset-offset ramps tested, suggesting that an increment on a pedestal is treated as a single auditory object whose detection is most difficult in the presence of other objects (in this case, ``beats'').

  15. Implications of dielectric breakdown weathering for the lunar regolith

    NASA Astrophysics Data System (ADS)

    Jordan, A.; Stubbs, T. J.; Wilson, J. K.; Schwadron, N.; Spence, H. E.; Izenberg, N.

    2015-12-01

    Solar energetic particles (SEPs) penetrate the lunar regolith to depths of ~1 mm and cause deep dielectric charging. The greatest charging occurs in permanently shadowed regions (PSRs), where temperatures make the electrical conductivity extremely low, which inhibits dissipating the charge buildup. Charging by very large SEP events may create subsurface electric fields that are strong enough to cause dielectric breakdown, or sparking, in the upper ~1 mm. Previous work has shown that, in PSRs, this breakdown weathering may have affected 10-25% of the meteoritically gardened regolith in PSRs and may thus be comparable to meteoritic weathering. But even regolith at lower latitudes can reach temperatures <100 K at night, causing it to dissipate charging in a few days--still enough to allow significant charging during large SEP events. Consequently, regolith at lower latitudes may also be susceptible to breakdown. We show how up to a few percent of gardened regolith at lower latitudes may have experienced breakdown. We also estimate the percentage of regolith that experienced breakdown during the two events detected in January and March 2012 by the Cosmic Ray Telescope for the Effects of Radiation (CRaTER) aboard the Lunar Reconnaissance Orbiter (LRO). Finally, we discuss what this more global view of breakdown weathering implies about the possibility of Apollo and Luna soil samples containing material that may have experienced breakdown.

  16. Observations of fast VHF-bright positive breakdown

    NASA Astrophysics Data System (ADS)

    Stock, M.; Krehbiel, P. R.; Rison, W.; Lapierre, J. L.; Edens, H. E.

    2014-12-01

    Positive breakdown during lightning discharges is generally considered to be weak and slowly propagating, as high speed video observations show it to be optically weak, and studies of the development of cloud-to-ground (CG) and intracloud (IC) flashes show development in the negative charge region to be slow. With the proper instrumentation, however, fast positive breakdown is a relatively common feature of both CG and IC flashes. The breakdown is bright at VHF, but is smoothly continuous so that time-of-arrival VHF mapping systems such as the Lightning Mapping Array are usually unable to detect or locate its occurrence. However, the breakdown is easily locatable using interferometric mapping techniques. Such an interferometer was developed at NM Tech in the 1980s and used in the CaPE studies at Kennedy Space Center in 1991, where it observed fast (1-6 × 107 m/s), VHF-bright positive leaders propagating away from the source region of negative CG return strokes (Shao et al., 1995). Here we report new observations of fast positive breakdown, obtained with Langmuir Laboratory's flash-continuous broadband VHF interferometer, that confirm and substantially expand our understanding of the phenomena. Numerous examples have been observed following return strokes of negative CG flashes, including bolt-from-blue discharges, and during K-processes of both IC and CG flashes. The breakdown typically propagates a few kilometers at speeds on the order of 107 m/s and frequently produces some of the brightest radiation of the flash. A particularly interesting feature of the breakdown is that it propagates into regions of previously un-ionized air. Then following the breakdown, frequently no further VHF emission is seen along or beyond its channel, indicating that the channel formed is not conducting. But on occasion, especially during cloud-to-ground flashes, the end of the fast positive breakdown turns into a normal, slowly propagating positive leader.

  17. Fundamental studies of passivity and passivity breakdown. Final report

    SciTech Connect

    Macdonald, D.D.; Urquidi-Macdonald, M.; Song, H.; Biaggio-Rocha, S.; Searson, P.

    1991-11-01

    This report summarizes the findings of our fundamental research program on passivity and passivity breakdown. During the past three and one half years in this program (including the three year incrementally-funded grant prior to the present grant), we developed and experimentally tested various physical models for the growth and breakdown of passive films on metal surfaces. These models belong to a general class termed ``point defects models`` (PDMs), in which the growth and breakdown of passive films are described in terms of the movement of anion and cation vacancies.

  18. Defining the Core Proteome of the Chloroplast Envelope Membranes

    PubMed Central

    Simm, Stefan; Papasotiriou, Dimitrios G.; Ibrahim, Mohamed; Leisegang, Matthias S.; Müller, Bernd; Schorge, Tobias; Karas, Michael; Mirus, Oliver; Sommer, Maik S.; Schleiff, Enrico

    2013-01-01

    High-throughput protein localization studies require multiple strategies. Mass spectrometric analysis of defined cellular fractions is one of the complementary approaches to a diverse array of cell biological methods. In recent years, the protein content of different cellular (sub-)compartments was approached. Despite of all the efforts made, the analysis of membrane fractions remains difficult, in that the dissection of the proteomes of the envelope membranes of chloroplasts or mitochondria is often not reliable because sample purity is not always warranted. Moreover, proteomic studies are often restricted to single (model) species, and therefore limited in respect to differential individual evolution. In this study we analyzed the chloroplast envelope proteomes of different plant species, namely, the individual proteomes of inner and outer envelope (OE) membrane of Pisum sativum and the mixed envelope proteomes of Arabidopsis thaliana and Medicago sativa. The analysis of all three species yielded 341 identified proteins in total, 247 of them being unique. 39 proteins were genuine envelope proteins found in at least two species. Based on this and previous envelope studies we defined the core envelope proteome of chloroplasts. Comparing the general overlap of the available six independent studies (including ours) revealed only a number of 27 envelope proteins. Depending on the stringency of applied selection criteria we found 231 envelope proteins, while less stringent criteria increases this number to 649 putative envelope proteins. Based on the latter we provide a map of the outer and inner envelope core proteome, which includes many yet uncharacterized proteins predicted to be involved in transport, signaling, and response. Furthermore, a foundation for the functional characterization of yet unidentified functions of the inner and OE for further analyses is provided. PMID:23390424

  19. NUCLEAR MEMBRANES FROM MAMMALIAN LIVER

    PubMed Central

    Franke, Werner W.; Deumling, Barbara; Ermen, Baerbel; Jarasch, Ernst-Dieter; Kleinig, Hans

    1970-01-01

    Nuclear membranes were isolated from rat and pig liver by sonication of highly purified nuclear fractions and subsequent removal of adhering nucleoproteins in a high salt medium. The fractions were examined in the electron microscope by both negative staining and thin sectioning techniques and were found to consist of nuclear envelope fragments of widely varying sizes. Nuclear pore complex constituents still could frequently be recognized. The chemical composition of the nuclear membrane fractions was determined and compared with those of microsomal fractions prepared in parallel. For total nuclei as well as for nuclear membranes and microsomes, various enzyme activities were studied. The results indicate that a similarity exists between both fractions of cytomembranes, nuclear envelope, and endoplasmic reticulum, with respect to their RNA:protein ratio and their content of polar and nonpolar lipids. Both membranous fractions had many proteins in common including some membrane-bound enzymes. Activities in Mg-ATPase and the two examined cytochrome reductases were of the same order of magnitude. The content of cytochrome b5 as well as of P-450 was markedly lower in the nuclear membranes. The nuclear membranes were found to have a higher buoyant density and to be richer in protein. The glucose-6-phosphatase and Na-K-ATPase activities in the nuclear membrane fraction were very low. In the gel electrophoresis, in addition to many common protein bands, some characteristic ones for either microsomal or nuclear membranous material were detected. Significant small amounts of DNA and RNA were found to remain closely associated with the nuclear envelope fragments. Our findings indicate that nuclear and endoplasmic reticulum membranes which are known to be in morphological continuity have, besides a far-reaching similarity, some characteristic differences. PMID:4317731

  20. Electrical Breakdown in a Martian Gas Mixture

    NASA Technical Reports Server (NTRS)

    Buhler, C. R.; Calle, C. I.; Nelson, E.

    2003-01-01

    The high probability for dust interactions during Martian dust storms and dust devils combined with the cold, dry climate of Mars most likely result in airborne dust that is highly charged. On Earth, potential gradients up to 5 kV/m have been recorded and in some cases resulted in lightning. Although the Martian atmosphere is not conducive to lightning generation, it is widely believed that electrical discharge in the form of a corona occurs. In order to understand the breakdown of gases, Paschen measurements are taken which relate the minimum potential required to spark across a gap between two electrodes. The minimum potential is plotted versus the pressure-distance value for electrodes of a given geometry. For most gases, the potential decreases as the pressure decreases. For CO2, the minimum in the curve happens to be at Mars atmospheric pressures (5-7 mm Hg) for many distances and geometries. However, a very small amount (<0.1%) of mixing gases radically changes the curve, as noted by Leach. Here, we present the first experimental results of a Paschen curve for a Mars gas mixture compared with 100% pure CO2.

  1. Rupture models with dynamically determined breakdown displacement

    USGS Publications Warehouse

    Andrews, D.J.

    2004-01-01

    The critical breakdown displacement, Dc, in which friction drops to its sliding value, can be made dependent on event size by specifying friction to be a function of variables other than slip. Two such friction laws are examined here. The first is designed to achieve accuracy and smoothness in discrete numerical calculations. Consistent resolution throughout an evolving rupture is achieved by specifying friction as a function of elapsed time after peak stress is reached. Such a time-weakening model produces Dc and fracture energy proportional to the square root of distance rupture has propagated in the case of uniform stress drop. The second friction law is more physically motivated. Energy loss in a damage zone outside the slip zone has the effect of increasing Dc and limiting peak slip velocity (Andrews, 1976). This article demonstrates a converse effect, that artificially limiting slip velocity on a fault in an elastic medium has a toughening effect, increasing fracture energy and Dc proportionally to rupture propagation distance in the case of uniform stress drop. Both the time-weakening and the velocity-toughening models can be used in calculations with heterogeneous stress drop.

  2. Magnetic breakdown in double quantum wells

    SciTech Connect

    Harff, N.E. |; Simmons, J.A.; Klem, J.F.; Boebinger, G.S.; Pfeiffer, L.N.; West, K.W.

    1996-08-01

    The authors find that a sufficiently large perpendicular magnetic field (B{sub {perpendicular}}) causes magnetic breakdown (MB) in coupled double quantum wells (QWs) that are subject to an in-plane magnetic field (B{sub {parallel}}). B{sub {parallel}} shifts one QW dispersion curve with respect to that of the other QW, resulting in an anticrossing and an energy gap. When the gap is below the Fermi level the resulting Fermi surface (FS) consists of two components, a lens-shaped inner orbit and an hour-glass shaped outer orbit. B{sub {perpendicular}} causes Landau level formation and Shubnikov-de Haas (SdH) oscillations for each component of the FS. MB occurs when the magnetic forces from B{sub {perpendicular}} become dominant and the electrons move on free-electron circular orbits rather than on the lens and hour-glass orbits. MB is observed by identifying the peaks present in the Fourier power spectrum of the longitudinal resistance vs. 1/B{sub {perpendicular}} at constant B{sub {parallel}}, an arrangement achieved with an in-situ tilting sample holder. Results are presented for two strongly coupled GaAs/AlGaAs DQW samples.

  3. Nanopore fabrication by controlled dielectric breakdown.

    PubMed

    Kwok, Harold; Briggs, Kyle; Tabard-Cossa, Vincent

    2014-01-01

    Nanofabrication techniques for achieving dimensional control at the nanometer scale are generally equipment-intensive and time-consuming. The use of energetic beams of electrons or ions has placed the fabrication of nanopores in thin solid-state membranes within reach of some academic laboratories, yet these tools are not accessible to many researchers and are poorly suited for mass-production. Here we describe a fast and simple approach for fabricating a single nanopore down to 2-nm in size with sub-nm precision, directly in solution, by controlling dielectric breakdown at the nanoscale. The method relies on applying a voltage across an insulating membrane to generate a high electric field, while monitoring the induced leakage current. We show that nanopores fabricated by this method produce clear electrical signals from translocating DNA molecules. Considering the tremendous reduction in complexity and cost, we envision this fabrication strategy would not only benefit researchers from the physical and life sciences interested in gaining reliable access to solid-state nanopores, but may provide a path towards manufacturing of nanopore-based biotechnologies.

  4. Medical Applications of Laser Induced Breakdown Spectroscopy

    NASA Astrophysics Data System (ADS)

    Pathak, A. K.; Rai, N. K.; Singh, Ankita; Rai, A. K.; Rai, Pradeep K.; Rai, Pramod K.

    2014-11-01

    Sedentary lifestyle of human beings has resulted in various diseases and in turn we require a potential tool that can be used to address various issues related to human health. Laser Induced Breakdown Spectroscopy (LIBS) is one such potential optical analytical tool that has become quite popular because of its distinctive features that include applicability to any type/phase of samples with almost no sample preparation. Several reports are available that discusses the capabilities of LIBS, suitable for various applications in different branches of science which cannot be addressed by traditional analytical methods but only few reports are available for the medical applications of LIBS. In the present work, LIBS has been implemented to understand the role of various elements in the formation of gallstones (formed under the empyema and mucocele state of gallbladder) samples along with patient history that were collected from Purvancal region of Uttar Pradesh, India. The occurrence statistics of gallstones under the present study reveal higher occurrence of gallstones in female patients. The gallstone occurrence was found more prevalent for those male patients who were having the habit of either tobacco chewing, smoking or drinking alcohols. This work further reports in-situ LIBS study of deciduous tooth and in-vivo LIBS study of human nail.

  5. Novel laser breakdown spectrometer for environmental monitoring

    NASA Astrophysics Data System (ADS)

    Mirov, Sergey B.; Pitt, Robert E.; Dergachev, Alex Y.; Lee, Wonwoo; Martyshkin, Dmitri V.; Mirov, Olga D.; Randolph, Jeremy J.; DeLucas, Lawrence J.; Brouillette, Christie G.; Basiev, Tasoltan T.; Orlovskii, Yurii V.; Alimov, Olimkhon K.; Vorob'ev, Ivan N.

    1999-11-01

    A novel experimental set-up using laser-induced breakdown spectroscopy (LIBS) for environmental analyses of heavy metals is described in this paper. It is based on state-of-the-art spectroscopic equipment, advanced detectors, and laser atomizers: a 0.75 m spectrometer ARC-750, intensified TE- cooled 256 X 1024 CCD camera, probe with fiber optic guide for signal transportation, and Nd:YAG laser plasma atomizers with two different methods for sample delivery. In the first method the liquid solution containing the atoms to be investigated is drawn into the chamber of the nebulizer. The mixture passes through the nozzle, accompanied by argon gas along with formed aerosol, and enters the plasma plume, which is generated by the laser spark in argon. The second method is based on direct generating of the plasma in the water jet of a continuously circulating sample. LIBS testing of samples containing Al, Cd, Cu, Fe, Pb, Zn, and Cr ions was compared with results using atomic absorption spectrophotometry. Initial indications showed good agreement between these two methods. Detection levels of less than 100 ppb were observed for copper and chromium. The described spectroscopic system exhibits high sensitivity, accumulation of luminescence spectrum in real time; and high dynamic range for concentrations detection from 100 ppb to 1000 ppm.

  6. Nuclear translocation of phospholipase C-zeta, an egg-activating factor, during early embryonic development

    SciTech Connect

    Sone, Yoshie; Ito, Masahiko; Shirakawa, Hideki; Shikano, Tomohide; Takeuchi, Hiroyuki; Kinoshita, Katsuyuki; Miyazaki, Shunichi . E-mail: shunm@research.twmu.ac.jp

    2005-05-13

    Phospholipase C-zeta (PLC{zeta}), a strong candidate of the egg-activating sperm factor, causes intracellular Ca{sup 2+} oscillations and egg activation, and is subsequently accumulated into the pronucleus (PN), when expressed in mouse eggs by injection of RNA encoding PLC{zeta}. Changes in the localization of expressed PLC{zeta} were investigated by tagging with a fluorescent protein. PLC{zeta} began to translocate into the PN formed at 5-6 h after RNA injection and increased there. Observation in the same embryo revealed that PLC{zeta} in the PN dispersed to the cytoplasm upon nuclear envelope breakdown and translocated again into the nucleus after cleavage. The dynamics was found in the second mitosis as well. When RNA was injected into fertilization-originated 1-cell embryos or blastomere(s) of 2-8-cell embryos, the nuclear localization of expressed PLC{zeta} was recognized in every embryo up to blastocyst. Thus, PLC{zeta} exhibited alternative cytoplasm/nucleus localization during development. This supports the view that the sperm factor could control cell cycle-dependent generation of Ca{sup 2+} oscillations in early embryogenesis.

  7. Fullerenes and fulleranes in circumstellar envelopes

    NASA Astrophysics Data System (ADS)

    Zhang, Yong; Kwok, Sun; Sadjadi, SeyedAbdolreza

    2016-07-01

    Three decades of search have recently led to convincing discoveries of cosmic fullerenes. The presence of C60 and C+ 60 in both circumstellar and interstellar environments suggests that these molecules and their derivatives can be efficiently formed in circumstellar envelopes and survive in harsh conditions. Detailed analysis of the infrared bands from fullerenes and their connections with the local properties can provide valuable information on the physical conditions and chemical processes that occurred in the late stages of stellar evolution. The identification of C+ 60 as the carrier of four diffuse interstellar bands (DIBs) suggests that fullerene- related compounds are abundant in interstellar space and are essential for resolving the DIB mystery. Experiments have revealed a high hydrogenation rate when C60 is exposed to atomic hydrogen, motivating the attempt to search for cosmic fulleranes. In this paper, we present a short review of current knowledge of cosmic fullerenes and fulleranes and briefly discuss the implications on circumstellar chemistry.

  8. Antireflection Pyrex envelopes for parabolic solar collectors

    NASA Astrophysics Data System (ADS)

    McCollister, H. L.; Pettit, R. B.

    1983-11-01

    Antireflective (AR) coatings, applied to the glass envelopes used in parabolic trough solar collectors around the receiver tube in order to reduce thermal losses, can increase solar transmittance by 7 percent. An AR surface has been formed on Pyrex by first heat treating the glass to cause a compositional phase separation, removing a surface layer after heat treatment through the use of a preetching solution, and finally etching in a solution that contains hydrofluorosilic and ammonium bifluoride acids. AR-coated samples with solar transmittance values of more than 0.97, by comparison to an untreated sample value of 0.91, have been obtained for the 560-630 C range of heat treatment temperatures. Optimum values have also been determined for the other processing parameters.

  9. A sensitive line search in circumstellar envelopes

    NASA Astrophysics Data System (ADS)

    Nguyen-Q-Rieu; Deguchi, S.; Izumiura, H.; Kaifu, N.; Ohishi, M.; Suzuki, H.; Ukita, N.

    A molecular line search in the range between 85 and 89 GHz has been performed in the circumstellar envelopes of 11 evolved stars. Emissions of 29SiO J=2-1,28SiO J=2-1, HCN J=1-0, H13CN J=1-0, HC5 N J=33-32, HCO+ J=1-0 transitions and other transitions of C2 H, C4 H, and C3 N have been observed in 11 stars. We have detected the ground state 29SiO J=2-1 maser in several stars. We have also detected HCN emission in VY CMa. A narrow H13CN spike feature near the central velocity has been found in the spectrum of CRL 2688.

  10. Beam-envelope theory of ionization cooling

    NASA Astrophysics Data System (ADS)

    Wang, Chun-xi; Kim, Kwang-Je

    2004-10-01

    Linear beam-envelope theory of ionization cooling in 6D phase space has been systematically established in the past few years. In this paper, we briefly review the general formalism as well as the specific theories for a quadrupole channel and a bent-solenoidal channel with symmetric focusing. These channels play important roles in the design of cooling channels for the envisioned neutrino factories and muon colliders. The analytical solutions of these channels are relatively simple yet provide good understanding of cooling and heating mechanisms in both transverse and longitudinal phase spaces. Furthermore, the resulting formulae can be used to evaluate cooling channel designs the same way as the radiation integrals are used in storage ring designs.

  11. Diversity in the fertilization envelopes of echinoderms.

    PubMed

    Oulhen, Nathalie; Reich, Adrian; Wong, Julian L; Ramos, Isabela; Wessel, Gary M

    2013-01-01

    Cell surface changes in an egg at fertilization are essential to begin development and for protecting the zygote. Most fertilized eggs construct a barrier around themselves by modifying their original extracellular matrix. This construction usually results from calcium-induced exocytosis of cortical granules, the contents of which in sea urchins function to form the fertilization envelope (FE), an extracellular matrix of cortical granule contents built upon a vitelline layer scaffold. Here, we examined the molecular mechanism of this process in sea stars, a close relative of the sea urchins, and analyze the evolutionary changes that likely occurred in the functionality of this structure between these two organisms. We find that the FE of sea stars is more permeable than in sea urchins, allowing diffusion of molecules in excess of 2 megadaltons. Through a proteomic and transcriptomic approach, we find that most, but not all, of the proteins present in the sea urchin envelope are present in sea stars, including SFE9, proteoliaisin, and rendezvin. The mRNAs encoding these FE proteins accumulated most densely in early oocytes, and then beginning with vitellogenesis, these mRNAs decreased in abundance to levels nearly undetectable in eggs. Antibodies to the SFE9 protein of sea stars showed that the cortical granules in sea star also accumulated most significantly in early oocytes, but different from sea urchins, they translocated to the cortex of the oocytes well before meiotic initiation. These results suggest that the preparation for cell surface changes in sea urchins has been shifted to later in oogenesis, and perhaps reflects the meiotic differences among the species-sea star oocytes are stored in prophase of meiosis and fertilized during the meiotic divisions, as in most animals, whereas sea urchins are one of the few taxons in which eggs have completed meiosis prior to fertilization.

  12. ASYMMETRIC ACCRETION FLOWS WITHIN A COMMON ENVELOPE

    SciTech Connect

    MacLeod, Morgan; Ramirez-Ruiz, Enrico

    2015-04-10

    This paper examines flows in the immediate vicinity of stars and compact objects dynamically inspiralling within a common envelope (CE). Flow in the vicinity of the embedded object is gravitationally focused, leading to drag and potentially to gas accretion. This process has been studied numerically and analytically in the context of Hoyle–Lyttleton accretion (HLA). Yet, within a CE, accretion structures may span a large fraction of the envelope radius, and in so doing sweep across a substantial radial gradient of density. We quantify these gradients using detailed stellar evolution models for a range of CE encounters. We provide estimates of typical scales in CE encounters that involve main sequence stars, white dwarfs, neutron stars, and black holes with giant-branch companions of a wide range of masses. We apply these typical scales to hydrodynamic simulations of three-dimensional HLA with an upstream density gradient. This density gradient breaks the symmetry that defines HLA flow, and imposes an angular momentum barrier to accretion. Material that is focused into the vicinity of the embedded object thus may not be able to accrete. As a result, accretion rates drop dramatically, by one to two orders of magnitude, while drag rates are only mildly affected. We provide fitting formulae to the numerically derived rates of drag and accretion as a function of the density gradient. The reduced ratio of accretion to drag suggests that objects that can efficiently gain mass during CE evolution, such as black holes and neutron stars, may grow less than implied by the HLA formalism.

  13. 10 CFR 434.516 - Building exterior envelope.

    Code of Federal Regulations, 2012 CFR

    2012-01-01

    ... 10 Energy 3 2012-01-01 2012-01-01 false Building exterior envelope. 434.516 Section 434.516 Energy DEPARTMENT OF ENERGY ENERGY CONSERVATION ENERGY CODE FOR NEW FEDERAL COMMERCIAL AND MULTI-FAMILY HIGH RISE RESIDENTIAL BUILDINGS Building Energy Cost Compliance Alternative § 434.516 Building exterior envelope....

  14. 10 CFR 434.516 - Building exterior envelope.

    Code of Federal Regulations, 2013 CFR

    2013-01-01

    ... 10 Energy 3 2013-01-01 2013-01-01 false Building exterior envelope. 434.516 Section 434.516 Energy DEPARTMENT OF ENERGY ENERGY CONSERVATION ENERGY CODE FOR NEW FEDERAL COMMERCIAL AND MULTI-FAMILY HIGH RISE RESIDENTIAL BUILDINGS Building Energy Cost Compliance Alternative § 434.516 Building exterior envelope....

  15. 10 CFR 434.516 - Building exterior envelope.

    Code of Federal Regulations, 2014 CFR

    2014-01-01

    ... 10 Energy 3 2014-01-01 2014-01-01 false Building exterior envelope. 434.516 Section 434.516 Energy DEPARTMENT OF ENERGY ENERGY CONSERVATION ENERGY CODE FOR NEW FEDERAL COMMERCIAL AND MULTI-FAMILY HIGH RISE RESIDENTIAL BUILDINGS Building Energy Cost Compliance Alternative § 434.516 Building exterior envelope....

  16. 14 CFR 29.1517 - Limiting height-speed envelope.

    Code of Federal Regulations, 2011 CFR

    2011-01-01

    ... 14 Aeronautics and Space 1 2011-01-01 2011-01-01 false Limiting height-speed envelope. 29.1517... Operating Limitations § 29.1517 Limiting height-speed envelope. For Category A rotorcraft, if a range of heights exists at any speed, including zero, within which it is not possible to make a safe...

  17. 14 CFR 27.87 - Height-speed envelope.

    Code of Federal Regulations, 2011 CFR

    2011-01-01

    ... 14 Aeronautics and Space 1 2011-01-01 2011-01-01 false Height-speed envelope. 27.87 Section 27.87... STANDARDS: NORMAL CATEGORY ROTORCRAFT Flight Performance § 27.87 Height-speed envelope. (a) If there is any combination of height and forward speed (including hover) under which a safe landing cannot be made under...

  18. 14 CFR 27.87 - Height-speed envelope.

    Code of Federal Regulations, 2013 CFR

    2013-01-01

    ... 14 Aeronautics and Space 1 2013-01-01 2013-01-01 false Height-speed envelope. 27.87 Section 27.87... STANDARDS: NORMAL CATEGORY ROTORCRAFT Flight Performance § 27.87 Height-speed envelope. (a) If there is any combination of height and forward speed (including hover) under which a safe landing cannot be made under...

  19. 14 CFR 29.1517 - Limiting height-speed envelope.

    Code of Federal Regulations, 2012 CFR

    2012-01-01

    ... 14 Aeronautics and Space 1 2012-01-01 2012-01-01 false Limiting height-speed envelope. 29.1517... Operating Limitations § 29.1517 Limiting height-speed envelope. For Category A rotorcraft, if a range of heights exists at any speed, including zero, within which it is not possible to make a safe...

  20. 14 CFR 29.1517 - Limiting height-speed envelope.

    Code of Federal Regulations, 2010 CFR

    2010-01-01

    ... 14 Aeronautics and Space 1 2010-01-01 2010-01-01 false Limiting height-speed envelope. 29.1517... Operating Limitations § 29.1517 Limiting height-speed envelope. For Category A rotorcraft, if a range of heights exists at any speed, including zero, within which it is not possible to make a safe...

  1. 14 CFR 29.1517 - Limiting height-speed envelope.

    Code of Federal Regulations, 2013 CFR

    2013-01-01

    ... 14 Aeronautics and Space 1 2013-01-01 2013-01-01 false Limiting height-speed envelope. 29.1517... Operating Limitations § 29.1517 Limiting height-speed envelope. For Category A rotorcraft, if a range of heights exists at any speed, including zero, within which it is not possible to make a safe...

  2. 14 CFR 27.87 - Height-speed envelope.

    Code of Federal Regulations, 2010 CFR

    2010-01-01

    ... 14 Aeronautics and Space 1 2010-01-01 2010-01-01 false Height-speed envelope. 27.87 Section 27.87... STANDARDS: NORMAL CATEGORY ROTORCRAFT Flight Performance § 27.87 Height-speed envelope. (a) If there is any combination of height and forward speed (including hover) under which a safe landing cannot be made under...

  3. 14 CFR 29.1517 - Limiting height-speed envelope.

    Code of Federal Regulations, 2014 CFR

    2014-01-01

    ... 14 Aeronautics and Space 1 2014-01-01 2014-01-01 false Limiting height-speed envelope. 29.1517... Operating Limitations § 29.1517 Limiting height-speed envelope. For Category A rotorcraft, if a range of heights exists at any speed, including zero, within which it is not possible to make a safe...

  4. 14 CFR 27.87 - Height-speed envelope.

    Code of Federal Regulations, 2014 CFR

    2014-01-01

    ... 14 Aeronautics and Space 1 2014-01-01 2014-01-01 false Height-speed envelope. 27.87 Section 27.87... STANDARDS: NORMAL CATEGORY ROTORCRAFT Flight Performance § 27.87 Height-speed envelope. (a) If there is any combination of height and forward speed (including hover) under which a safe landing cannot be made under...

  5. 14 CFR 27.87 - Height-speed envelope.

    Code of Federal Regulations, 2012 CFR

    2012-01-01

    ... 14 Aeronautics and Space 1 2012-01-01 2012-01-01 false Height-speed envelope. 27.87 Section 27.87... STANDARDS: NORMAL CATEGORY ROTORCRAFT Flight Performance § 27.87 Height-speed envelope. (a) If there is any combination of height and forward speed (including hover) under which a safe landing cannot be made under...

  6. 14 CFR 25.333 - Flight maneuvering envelope.

    Code of Federal Regulations, 2014 CFR

    2014-01-01

    ... 14 Aeronautics and Space 1 2014-01-01 2014-01-01 false Flight maneuvering envelope. 25.333 Section... AIRWORTHINESS STANDARDS: TRANSPORT CATEGORY AIRPLANES Structure Flight Maneuver and Gust Conditions § 25.333 Flight maneuvering envelope. (a) General. The strength requirements must be met at each combination...

  7. 14 CFR 29.87 - Height-velocity envelope.

    Code of Federal Regulations, 2014 CFR

    2014-01-01

    ... 14 Aeronautics and Space 1 2014-01-01 2014-01-01 false Height-velocity envelope. 29.87 Section 29... AIRWORTHINESS STANDARDS: TRANSPORT CATEGORY ROTORCRAFT Flight Performance § 29.87 Height-velocity envelope. (a) If there is any combination of height and forward velocity (including hover) under which a...

  8. 14 CFR 29.87 - Height-velocity envelope.

    Code of Federal Regulations, 2011 CFR

    2011-01-01

    ... 14 Aeronautics and Space 1 2011-01-01 2011-01-01 false Height-velocity envelope. 29.87 Section 29... AIRWORTHINESS STANDARDS: TRANSPORT CATEGORY ROTORCRAFT Flight Performance § 29.87 Height-velocity envelope. (a) If there is any combination of height and forward velocity (including hover) under which a...

  9. 14 CFR 29.87 - Height-velocity envelope.

    Code of Federal Regulations, 2013 CFR

    2013-01-01

    ... 14 Aeronautics and Space 1 2013-01-01 2013-01-01 false Height-velocity envelope. 29.87 Section 29... AIRWORTHINESS STANDARDS: TRANSPORT CATEGORY ROTORCRAFT Flight Performance § 29.87 Height-velocity envelope. (a) If there is any combination of height and forward velocity (including hover) under which a...

  10. 14 CFR 29.87 - Height-velocity envelope.

    Code of Federal Regulations, 2010 CFR

    2010-01-01

    ... 14 Aeronautics and Space 1 2010-01-01 2010-01-01 false Height-velocity envelope. 29.87 Section 29... AIRWORTHINESS STANDARDS: TRANSPORT CATEGORY ROTORCRAFT Flight Performance § 29.87 Height-velocity envelope. (a) If there is any combination of height and forward velocity (including hover) under which a...

  11. 14 CFR 29.87 - Height-velocity envelope.

    Code of Federal Regulations, 2012 CFR

    2012-01-01

    ... 14 Aeronautics and Space 1 2012-01-01 2012-01-01 false Height-velocity envelope. 29.87 Section 29... AIRWORTHINESS STANDARDS: TRANSPORT CATEGORY ROTORCRAFT Flight Performance § 29.87 Height-velocity envelope. (a) If there is any combination of height and forward velocity (including hover) under which a...

  12. Data Envelopment Analysis: Measurement of Educational Efficiency in Texas

    ERIC Educational Resources Information Center

    Carter, Lacy

    2012-01-01

    The purpose of this study was to examine the efficiency of Texas public school districts through Data Envelopment Analysis. The Data Envelopment Analysis estimation method calculated and assigned efficiency scores to each of the 931 school districts considered in the study. The efficiency scores were utilized in two phases. First, the school…

  13. When Lamins Go Bad: Nuclear Structure and Disease

    PubMed Central

    Schreiber, Katherine H.; Kennedy, Brian K.

    2013-01-01

    When mutations in nuclear lamins were first identified in skeletal and cardiac muscle diseases, the molecular events underlying pathogenesis were mere points of speculation. As more and more unrelated diseases were linked to lamins and other nuclear envelope proteins, nuclear structure and disease became an increasingly prominent research focus. Today, the disease mechanisms remain unresolved, but incredible progress has occurred. Nuclear envelope dysfunction is not only associated with altered nuclear activity, but also impaired structural dynamics and aberrant cell signaling. Building on these findings, small molecules are being discovered in animal models that may become effective therapeutic agents. PMID:23498943

  14. Detection of uranium using laser-induced breakdown spectroscopy.

    PubMed

    Chinni, Rosemarie C; Cremers, David A; Radziemski, Leon J; Bostian, Melissa; Navarro-Northrup, Claudia

    2009-11-01

    The goal of this work is a detailed study of uranium detection by laser-induced breakdown spectroscopy (LIBS) for application to activities associated with environmental surveillance and detecting weapons of mass destruction (WMD). The study was used to assist development of LIBS instruments for standoff detection of bulk radiological and nuclear materials and these materials distributed as contaminants on surfaces. Uranium spectra were analyzed under a variety of different conditions at room pressure, reduced pressures, and in an argon atmosphere. All spectra displayed a high apparent background due to the high density of uranium lines. Time decay curves of selected uranium lines were monitored and compared to other elements in an attempt to maximize detection capabilities for each species in the complicated uranium spectrum. A survey of the LIBS uranium spectra was conducted and relative emission line strengths were determined over the range of 260 to 800 nm. These spectra provide a guide for selection of the strongest LIBS analytical lines for uranium detection in different spectral regions. A detection limit for uranium in soil of 0.26% w/w was obtained at close range and 0.5% w/w was achieved at a distance of 30 m. Surface detection limits were substrate dependent and ranged from 13 to 150 microg/cm2. Double-pulse experiments (both collinear and orthogonal arrangements) were shown to enhance the uranium signal in some cases. Based on the results of this work, a short critique is given of the applicability of LIBS for the detection of uranium residues on surfaces for environmental monitoring and WMD surveillance. PMID:19891832

  15. Detection of uranium using laser-induced breakdown spectroscopy.

    PubMed

    Chinni, Rosemarie C; Cremers, David A; Radziemski, Leon J; Bostian, Melissa; Navarro-Northrup, Claudia

    2009-11-01

    The goal of this work is a detailed study of uranium detection by laser-induced breakdown spectroscopy (LIBS) for application to activities associated with environmental surveillance and detecting weapons of mass destruction (WMD). The study was used to assist development of LIBS instruments for standoff detection of bulk radiological and nuclear materials and these materials distributed as contaminants on surfaces. Uranium spectra were analyzed under a variety of different conditions at room pressure, reduced pressures, and in an argon atmosphere. All spectra displayed a high apparent background due to the high density of uranium lines. Time decay curves of selected uranium lines were monitored and compared to other elements in an attempt to maximize detection capabilities for each species in the complicated uranium spectrum. A survey of the LIBS uranium spectra was conducted and relative emission line strengths were determined over the range of 260 to 800 nm. These spectra provide a guide for selection of the strongest LIBS analytical lines for uranium detection in different spectral regions. A detection limit for uranium in soil of 0.26% w/w was obtained at close range and 0.5% w/w was achieved at a distance of 30 m. Surface detection limits were substrate dependent and ranged from 13 to 150 microg/cm2. Double-pulse experiments (both collinear and orthogonal arrangements) were shown to enhance the uranium signal in some cases. Based on the results of this work, a short critique is given of the applicability of LIBS for the detection of uranium residues on surfaces for environmental monitoring and WMD surveillance.

  16. D STAND DELIVERY END OF #44 TANDEM BREAKDOWN MILL WITH ...

    Library of Congress Historic Buildings Survey, Historic Engineering Record, Historic Landscapes Survey

    D STAND DELIVERY END OF #44 TANDEM BREAKDOWN MILL WITH UPCOILER. BACKUP ROLLS, 40 TONS. WORK ROLLS, 20 TONS., C. 1900. OPERATING SPEED, 600'/MINUTE. AUTOMATIC GAUGE CONTROL. - American Brass Foundry, 70 Sayre Street, Buffalo, Erie County, NY

  17. Nanosecond-gated laser induced breakdown spectroscopy in hydrocarbon mixtures

    NASA Astrophysics Data System (ADS)

    Kobayashi, Kazunobu; Bak, Moon Soo; Tanaka, Hiroki; Do, Hyungrok

    2015-09-01

    Nanosecond-gated laser induced breakdown spectroscopy have been carried out in four different hydrocarbon gas mixtures (CH4/CO2/O2/N2, C2H4/O2/N2, C3H8/CO2/O2/N2 and C4H10/CO2/O2/N2) to investigate the effect of gas species on the laser induced breakdown kinetics and resulting the plasma emission. For this purpose, each mixture that consists of different species has the same atom composition. It is found that the temporal emission spectra and the decay rates of atomic line-intensities are almost identical for the breakdowns in the four different mixtures. This finding may indicate that the breakdown plasmas of these mixtures reach a similar thermodynamic and physiochemical state after its formation, resulting in a similar trend of quenching of excited species.

  18. Suppressing the cellular breakdown in silicon supersaturated with titanium

    NASA Astrophysics Data System (ADS)

    Liu, Fang; Prucnal, S.; Hübner, R.; Yuan, Ye; Skorupa, W.; Helm, M.; Zhou, Shengqiang

    2016-06-01

    Hyper doping Si with up to 6 at.% Ti in solid solution was performed by ion implantation followed by pulsed laser annealing and flash lamp annealing. In both cases, the implanted Si layer can be well recrystallized by liquid phase epitaxy and solid phase epitaxy, respectively. Cross-sectional transmission electron microscopy of Ti-implanted Si after liquid phase epitaxy shows the so-called growth interface breakdown or cellular breakdown owing to the occurrence of constitutional supercooling in the melt. The appearance of cellular breakdown prevents further recrystallization. However, the out-diffusion and cellular breakdown can be effectively suppressed by solid phase epitaxy during flash lamp annealing due to the high velocity of amorphous-crystalline interface and the low diffusion velocity for Ti in the solid phase.

  19. Vortex breakdown in closed containers with polygonal cross sections

    SciTech Connect

    Naumov, I. V. Dvoynishnikov, S. V.; Kabardin, I. K.; Tsoy, M. A.

    2015-12-15

    The vortex breakdown bubble in the confined flow generated by a rotating lid in closed containers with polygonal cross sections was analysed both experimentally and numerically for the height/radius aspect ratio equal to 2. The stagnation point locations of the breakdown bubble emergence and the corresponding Reynolds number were determined experimentally and in addition computed numerically by STAR-CCM+ CFD software for square, pentagonal, hexagonal, and octagonal cross section configurations. The flow pattern and the velocity were observed and measured by combining the seeding particle visualization and the temporal accuracy of laser Doppler anemometry. The vortex breakdown size and position on the container axis were determined for Reynolds numbers, ranging from 1450 to 2400. The obtained results were compared with the flow structure in the closed container of cubical and cylindrical configurations. It is shown that the measured evolution of steady vortex breakdown is in close agreement with the numerical results.

  20. Vortex breakdown in closed containers with polygonal cross sections

    NASA Astrophysics Data System (ADS)

    Naumov, I. V.; Dvoynishnikov, S. V.; Kabardin, I. K.; Tsoy, M. A.

    2015-12-01

    The vortex breakdown bubble in the confined flow generated by a rotating lid in closed containers with polygonal cross sections was analysed both experimentally and numerically for the height/radius aspect ratio equal to 2. The stagnation point locations of the breakdown bubble emergence and the corresponding Reynolds number were determined experimentally and in addition computed numerically by STAR-CCM+ CFD software for square, pentagonal, hexagonal, and octagonal cross section configurations. The flow pattern and the velocity were observed and measured by combining the seeding particle visualization and the temporal accuracy of laser Doppler anemometry. The vortex breakdown size and position on the container axis were determined for Reynolds numbers, ranging from 1450 to 2400. The obtained results were compared with the flow structure in the closed container of cubical and cylindrical configurations. It is shown that the measured evolution of steady vortex breakdown is in close agreement with the numerical results.

  1. 19 CFR 141.87 - Breakdown on component materials.

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ... OF THE TREASURY (CONTINUED) ENTRY OF MERCHANDISE Invoices § 141.87 Breakdown on component materials. Whenever the classification or appraisement of merchandise depends on the component materials, the...

  2. 19 CFR 141.87 - Breakdown on component materials.

    Code of Federal Regulations, 2013 CFR

    2013-04-01

    ... OF THE TREASURY (CONTINUED) ENTRY OF MERCHANDISE Invoices § 141.87 Breakdown on component materials. Whenever the classification or appraisement of merchandise depends on the component materials, the...

  3. 19 CFR 141.87 - Breakdown on component materials.

    Code of Federal Regulations, 2011 CFR

    2011-04-01

    ... OF THE TREASURY (CONTINUED) ENTRY OF MERCHANDISE Invoices § 141.87 Breakdown on component materials. Whenever the classification or appraisement of merchandise depends on the component materials, the...

  4. 19 CFR 141.87 - Breakdown on component materials.

    Code of Federal Regulations, 2014 CFR

    2014-04-01

    ... OF THE TREASURY (CONTINUED) ENTRY OF MERCHANDISE Invoices § 141.87 Breakdown on component materials. Whenever the classification or appraisement of merchandise depends on the component materials, the...

  5. A Spectral Algorithm for Envelope Reduction of Sparse Matrices

    NASA Technical Reports Server (NTRS)

    Barnard, Stephen T.; Pothen, Alex; Simon, Horst D.

    1993-01-01

    The problem of reordering a sparse symmetric matrix to reduce its envelope size is considered. A new spectral algorithm for computing an envelope-reducing reordering is obtained by associating a Laplacian matrix with the given matrix and then sorting the components of a specified eigenvector of the Laplacian. This Laplacian eigenvector solves a continuous relaxation of a discrete problem related to envelope minimization called the minimum 2-sum problem. The permutation vector computed by the spectral algorithm is a closest permutation vector to the specified Laplacian eigenvector. Numerical results show that the new reordering algorithm usually computes smaller envelope sizes than those obtained from the current standard algorithms such as Gibbs-Poole-Stockmeyer (GPS) or SPARSPAK reverse Cuthill-McKee (RCM), in some cases reducing the envelope by more than a factor of two.

  6. Interferometric Imaging of Molecular Envelopes with and without YSOs<

    NASA Astrophysics Data System (ADS)

    Ohashi, Nagayoshi

    1999-10-01

    Molecular envelopes are sites of star formation, and their geometrical and kinematical properties are very important to understand star formation. Particularly, their velocity structures, such as infall or rotation, need to be studied in detail to understand processes essential for star-formation. In order to investigate the physical properties of molecular envelopes in very detail, we need fine angular and velocity resolutions, which resolve both geometrical and velocity structures of molecular envelopes. A millimeter & submillimeter-wave interferometer is a very powerful tool providing high angular and velocity resolutions. Interferometric observations have realized direct imaging of infalling motions in molecular envelopes. In my talk, I will review what we learned about the physical properties of molecular envelopes with and without young stellar objects (YSOs) through interferometric observations. I will also discuss what we may learn about star-formation using a large millimeter & submillimeter array.

  7. Is the Breakdown of the Born-Oppenheimer Approximation Responsible for Internal Conversion in Large Molecules?*

    PubMed Central

    Burland, D. M.; Robinson, G. W.

    1970-01-01

    Vibronic radiationless transitions in large polyatomic molecules can be thought of as a process whereby the molecule, initially prepared in a discrete quasistationary state, makes a transition to an adjoining vibronic continuum belonging to a lower electronic state of the same multiplicity. In many instances the transition is analogous to penetration through a barrier between two „nested” potential sheets far away from an actual intersection of the sheets. Simultaneous distortions of vibrational and electronic parts of the wavefunction are required for such a tunneling process. The vibrational distortion manifests itself in the familiar Franck-Condon effect. The electronic distortion can be caused by nontotally symmetric vibrations of the molecule either because of the displaced nuclear configuration, to which the electrons instantly respond (Herzberg-Teller effect), or because of the protracted response of the electrons to the kinetic energy of nuclear motion (nonadiabatic effect or the breakdown of the Born-Oppenheimer approximation). It is found here that Herzberg-Teller coupling is more effective in causing radiationless transitions than the breakdown of the Born-Oppenheimer approximation when the potential surfaces involved are not close to an intersection. PMID:16591837

  8. Nanopore formation by controlled electrical breakdown: Efficient molecular-sensors

    NASA Astrophysics Data System (ADS)

    Abdalla, S.; Al-Marzouki, F. M.; Abdel-Daiem, A. M.

    2016-08-01

    A controlled electrical breakdown is used to produce efficient nanopore (NP) sensors. This phenomenon can be used to precisely fabricate these nanopore (NP) sensors through the membranes of the polydimethylsiloxane microarrays. This can be carried out, when localizing the electrical potential through a suitable microfluidic channel. Organic molecules, and other different protein-molecules, can be easily and precisely detected using this procedure referred to as controlled electrical breakdown technique.

  9. Ionizing potential waves and high-voltage breakdown streamers.

    NASA Technical Reports Server (NTRS)

    Albright, N. W.; Tidman, D. A.

    1972-01-01

    The structure of ionizing potential waves driven by a strong electric field in a dense gas is discussed. Negative breakdown waves are found to propagate with a velocity proportional to the electric field normal to the wavefront. This causes a curved ionizing potential wavefront to focus down into a filamentary structure, and may provide the reason why breakdown in dense gases propagates in the form of a narrow leader streamer instead of a broad wavefront.

  10. Non-Nuclear Testing of Space Nuclear Systems at NASA MSFC

    NASA Technical Reports Server (NTRS)

    Houts, Michael G.; Pearson, Boise J.; Aschenbrenner, Kenneth C.; Bradley, David E.; Dickens, Ricky; Emrich, William J.; Garber, Anne; Godfroy, Thomas J.; Harper, Roger T.; Martin, Jim J.; Polzin, Kurt; Schoenfeld, Michael P.; Webster, Kenneth L.

    2010-01-01

    Highly realistic non-nuclear testing can be used to investigate and resolve potential issues with space nuclear power and propulsion systems. Non-nuclear testing is particularly useful for systems designed with fuels and materials operating within their demonstrated nuclear performance envelope. Non-nuclear testing allows thermal hydraulic, heat transfer, structural, integration, safety, operational, performance, and other potential issues to be investigated and resolved with a greater degree of flexibility and at reduced cost and schedule compared to nuclear testing. The primary limit of non-nuclear testing is that nuclear characteristics and potential nuclear issues cannot be directly investigated. However, non-nuclear testing can be used to augment the potential benefit from any nuclear testing that may be required for space nuclear system design and development. This paper describes previous and ongoing non-nuclear testing related to space nuclear systems at NASA's Marshall Space Flight Center (MSFC).

  11. Acoustics of laminar boundary layers breakdown

    NASA Astrophysics Data System (ADS)

    Wang, Meng

    1994-12-01

    Boundary layer flow transition has long been suggested as a potential noise source in both marine (sonar-dome self noise) and aeronautical (aircraft cabin noise) applications, owing to the highly transient nature of process. The design of effective noise control strategies relies upon a clear understanding of the source mechanisms associated with the unsteady flow dynamics during transition. Due to formidable mathematical difficulties, theoretical predictions either are limited to early linear and weakly nonlinear stages of transition, or employ acoustic analogy theories based on approximate source field data, often in the form of empirical correlation. In the present work, an approach which combines direct numerical simulation of the source field with the Lighthill acoustic analogy is utilized. This approach takes advantage of the recent advancement in computational capabilities to obtain detailed information about the flow-induced acoustic sources. The transitional boundary layer flow is computed by solving the incompressible Navier-Stokes equations without model assumptions, thus allowing a direct evaluation of the pseudosound as well as source functions, including the Lighthill stress tensor and the wall shear stress. The latter are used for calculating the radiated pressure field based on the Curle-Powell solution of the Lighthill equation. This procedure allows a quantitative assessment of noise source mechanisms and the associated radiation characteristics during transition from primary instability up to the laminar breakdown stage. In particular, one is interested in comparing the roles played by the fluctuating volume Reynolds stress and the wall-shear-stresses, and in identifying specific flow processes and structures that are effective noise generators.

  12. Acoustics of laminar boundary layers breakdown

    NASA Technical Reports Server (NTRS)

    Wang, Meng

    1994-01-01

    Boundary layer flow transition has long been suggested as a potential noise source in both marine (sonar-dome self noise) and aeronautical (aircraft cabin noise) applications, owing to the highly transient nature of process. The design of effective noise control strategies relies upon a clear understanding of the source mechanisms associated with the unsteady flow dynamics during transition. Due to formidable mathematical difficulties, theoretical predictions either are limited to early linear and weakly nonlinear stages of transition, or employ acoustic analogy theories based on approximate source field data, often in the form of empirical correlation. In the present work, an approach which combines direct numerical simulation of the source field with the Lighthill acoustic analogy is utilized. This approach takes advantage of the recent advancement in computational capabilities to obtain detailed information about the flow-induced acoustic sources. The transitional boundary layer flow is computed by solving the incompressible Navier-Stokes equations without model assumptions, thus allowing a direct evaluation of the pseudosound as well as source functions, including the Lighthill stress tensor and the wall shear stress. The latter are used for calculating the radiated pressure field based on the Curle-Powell solution of the Lighthill equation. This procedure allows a quantitative assessment of noise source mechanisms and the associated radiation characteristics during transition from primary instability up to the laminar breakdown stage. In particular, one is interested in comparing the roles played by the fluctuating volume Reynolds stress and the wall-shear-stresses, and in identifying specific flow processes and structures that are effective noise generators.

  13. Breakdown of atmospheric pressure microgaps at high excitation frequencies

    NASA Astrophysics Data System (ADS)

    Levko, Dmitry; Raja, Laxminarayan

    2015-09-01

    Microwave breakdown of atmospheric pressure microgaps was studied by a one-dimensional Particle-in-Cell Monte Carlo Collisions numerical model. The effect of both field electron emission and secondary electron emission (due to electron impact, ion impact, and primary electron reflection) from surfaces on the breakdown process is considered. For conditions where field emission is the dominant electron emission mechanism from the electrode surfaces, it is found that the breakdown voltage of mw microdischarge coincides with the breakdown voltage of direct-current microdischarge. When microdischarge properties are controlled by both field and secondary electron emission, breakdown voltage of mw microdischarge exceeds that of dc microdischarge. When microdischarge is controlled only by secondary electron emission, breakdown voltage of mw microdischarge is smaller than that of dc microdischarge. It is shown that if the interelectrode gap exceeds some critical value, mw microdischarge can be ignited only by electrons initially seeded within the gap volume. In addition, the influence of electron reflection and secondary emission due to electron impact is studied. This work was supported by the Air Force Office of Scientific Research.

  14. Breakdown of atmospheric pressure microgaps at high excitation frequencies

    NASA Astrophysics Data System (ADS)

    Levko, Dmitry; Raja, Laxminarayan L.

    2015-05-01

    Microwave (mw) breakdown of atmospheric pressure microgaps is studied by a one-dimensional Particle-in-Cell Monte Carlo Collisions numerical model. The effect of both field electron emission and secondary electron emission (due to electron impact, ion impact, and primary electron reflection) from surfaces on the breakdown process is considered. For conditions where field emission is the dominant electron emission mechanism from the electrode surfaces, it is found that the breakdown voltage of mw microdischarge coincides with the breakdown voltage of direct-current (dc) microdischarge. When microdischarge properties are controlled by both field and secondary electron emission, breakdown voltage of mw microdischarge exceeds that of dc microdischarge. When microdischarge is controlled only by secondary electron emission, breakdown voltage of mw microdischarge is smaller than that of dc microdischarge. It is shown that if the interelectrode gap exceeds some critical value, mw microdischarge can be ignited only by electrons initially seeded within the gap volume. In addition, the influence of electron reflection and secondary emission due to electron impact is studied.

  15. AVLIS Production Plant work breakdown structure and Dictionary

    SciTech Connect

    Not Available

    1984-11-15

    The work breakdown structure has been prepared for the AVLIS Production Plant to define, organize, and identify the work efforts and is summarized in Fig. 1-1 for the top three project levels. The work breakdown structure itself is intended to be the primary organizational tool of the AVLIS Production Plant and is consistent with the overall AVLIS Program Work Breakdown Structure. It is designed to provide a framework for definition and accounting of all of the elements that are required for the eventual design, procurement, and construction of the AVLIS Production Plant. During the present phase of the AVLIS Project, the conceptual engineering phase, the work breakdown structure is intended to be the master structure and project organizer of documents, designs, and cost estimates. As the master project organizer, the key role of the work breakdown structure is to provide the mechanism for developing completeness in AVLIS cost estimates and design development of all hardware and systems. The work breakdown structure provides the framework for tracking, on a one-to-one basis, the component design criteria, systems requirements, design concepts, design drawings, performance projections, and conceptual cost estimates. It also serves as a vehicle for contract reporting. 12 figures, 2 tables.

  16. Breakdown of atmospheric pressure microgaps at high excitation frequencies

    SciTech Connect

    Levko, Dmitry; Raja, Laxminarayan L.

    2015-05-07

    Microwave (mw) breakdown of atmospheric pressure microgaps is studied by a one-dimensional Particle-in-Cell Monte Carlo Collisions numerical model. The effect of both field electron emission and secondary electron emission (due to electron impact, ion impact, and primary electron reflection) from surfaces on the breakdown process is considered. For conditions where field emission is the dominant electron emission mechanism from the electrode surfaces, it is found that the breakdown voltage of mw microdischarge coincides with the breakdown voltage of direct-current (dc) microdischarge. When microdischarge properties are controlled by both field and secondary electron emission, breakdown voltage of mw microdischarge exceeds that of dc microdischarge. When microdischarge is controlled only by secondary electron emission, breakdown voltage of mw microdischarge is smaller than that of dc microdischarge. It is shown that if the interelectrode gap exceeds some critical value, mw microdischarge can be ignited only by electrons initially seeded within the gap volume. In addition, the influence of electron reflection and secondary emission due to electron impact is studied.

  17. Decision Envelopment Analysis of space and terrestrially-based large scale commercial power systems for earth

    NASA Astrophysics Data System (ADS)

    Criswell, David R.; Thompson, Russell G.

    1992-08-01

    Decision Envelopment Analysis (DEA), the detailed quantitative comparison of alternative economic systems, is used to compare the technical efficiency of the large-scale power systems needed to meet the growing energy needs of terrestrial society. The Lunar Power System (LPS) captures sunlight on the moon, converts it to microwaves and beams the power to receivers on earth that output electricity. In terms of benefits versus costs, normalized to the range of 0 to 1, DEA reveals that LPS is at least ten times more efficient than conventional terrestrial solar-thermal and -photovoltaic, fossil, and nuclear systems. LPS is also environmentally benign compared to the conventional systems.

  18. Microbiological implications of electric field effects. II. Inactivation of yeast cells and repair of their cell envelope.

    PubMed

    Jacob, H E; Förster, W; Berg, H

    1981-01-01

    The inactivation of yeast cells in different growth phases by an electric field pulse was investigated. Cells of Saccharomyces cerevisiae in the logarithmic growth phase were found to be much more sensitive with respect to an electric discharge than those in the stationary phase. The influence of the electric field pulse characteristics on the inactivation as well as possible secondary effects were studied. The polyene antibiotic perhydrohexafungin (PHF) is used as a tool to sense defects in the yeast cell envelope brought about by electric field action. The repair kinetics of these defects was followed after the impulse. At least two repair stages can be distinguished, a fast one in the second range and a slower one which takes place after plating the cells on a nutrient medium. The obtained results are discussed in connection with current theories of reversible dielectric breakdown in biological membrane systems. PMID:7023081

  19. CLOSE STELLAR BINARY SYSTEMS BY GRAZING ENVELOPE EVOLUTION

    SciTech Connect

    Soker, Noam

    2015-02-20

    I suggest a spiral-in process in which a stellar companion grazes the envelope of a giant star while both the orbital separation and the giant radius shrink simultaneously, forming a close binary system. The binary system might be viewed as evolving in a constant state of 'just entering a common envelope (CE) phase.' In cases where this process takes place, it can be an alternative to CE evolution where the secondary star is immersed in the giant's envelope. Grazing envelope evolution (GEE) is made possible only if the companion manages to accrete mass at a high rate and launches jets that remove the outskirts of the giant envelope, hence preventing the formation of a CE. The high accretion rate is made possible by the accretion disk launching jets which efficiently carry the excess angular momentum and energy from the accreted mass. The orbital decay itself is caused by the gravitational interaction of the secondary star with the envelope inward of its orbit, i.e., dynamical friction (gravitational tide). Mass loss through the second Lagrangian point can carry additional angular momentum and envelope mass. The GEE lasts for tens to hundreds of years. The high accretion rate, with peaks lasting from months to years, might lead to a bright object referred to as the intermediate luminosity optical transient (Red Novae; Red Transients). A bipolar nebula and/or equatorial ring are formed around the binary remnant.

  20. Vitelline envelope, chorion, and micropyle of Fundulus heteroclitus eggs

    SciTech Connect

    Dumont, J.N.; Brummet, A.R.

    1980-01-01

    The architecture and transformation of the vitelline envelope of the developing oocyte into the chorion of the mature egg of Fundulus heteroclitus have been examined by scanning and transmission electron microscopy. The mature vitelline envelope is structurally complex and consists of about nine strata. The envelope is penetrated by pore canals that contain microvilli arising from the oocyte and macrovilli from follicle cells. During the envelope's transformation into the chorion, the pore canals are lost and the envelope becomes more fibrous and compact and its stratified nature less apparent. The micropyle, or pore, through which the sperm gains access to the enclosed egg is located at the bottom of a small funnel-shaped depression in the envelope. Internally, the micropyle opens on the apex of a cone-like elevation of the chorion. During the development of the envelope, structured chorionic fibrils, the components of which are presumed to be synthesized by the follicle cells, become attached to its surface. These chorionic fibrils are thought to aid in the attachment of the egg to the substratum and perhaps to help prevent water loss during low tides when the egg may be exposed.

  1. Neural coding of echo-envelope disparities in echolocating bats.

    PubMed

    Borina, Frank; Firzlaff, Uwe; Wiegrebe, Lutz

    2011-05-01

    The effective use of echolocation requires not only measuring the delay between the emitted call and returning echo to estimate the distance of an ensonified object. To locate an object in azimuth and elevation, the bat's auditory system must analyze the returning echoes in terms of their binaural properties, i.e., the echoes' interaural intensity and time differences (IIDs and ITDs). The effectiveness of IIDs for echolocation is undisputed, but when bats ensonify complex objects, the temporal structure of echoes may facilitate the analysis of the echo envelope in terms of envelope ITDs. Using extracellular recordings from the auditory midbrain of the bat, Phyllostomus discolor, we found a population of neurons that are sensitive to envelope ITDs of echoes of their sonar calls. Moreover, the envelope-ITD sensitivity improved with increasing temporal fluctuations in the echo envelopes, a sonar parameter related to the spatial statistics of complex natural reflectors like vegetation. The data show that in bats envelope ITDs may be used not only to locate external, prey-generated rustling sounds but also in the context of echolocation. Specifically, the temporal fluctuations in the echo envelope, which are created when the sonar emission is reflected from a complex natural target, support ITD-mediated echolocation.

  2. Aeroelastic Model Structure Computation for Envelope Expansion

    NASA Technical Reports Server (NTRS)

    Kukreja, Sunil L.

    2007-01-01

    Structure detection is a procedure for selecting a subset of candidate terms, from a full model description, that best describes the observed output. This is a necessary procedure to compute an efficient system description which may afford greater insight into the functionality of the system or a simpler controller design. Structure computation as a tool for black-box modeling may be of critical importance in the development of robust, parsimonious models for the flight-test community. Moreover, this approach may lead to efficient strategies for rapid envelope expansion that may save significant development time and costs. In this study, a least absolute shrinkage and selection operator (LASSO) technique is investigated for computing efficient model descriptions of non-linear aeroelastic systems. The LASSO minimises the residual sum of squares with the addition of an l(Sub 1) penalty term on the parameter vector of the traditional l(sub 2) minimisation problem. Its use for structure detection is a natural extension of this constrained minimisation approach to pseudo-linear regression problems which produces some model parameters that are exactly zero and, therefore, yields a parsimonious system description. Applicability of this technique for model structure computation for the F/A-18 (McDonnell Douglas, now The Boeing Company, Chicago, Illinois) Active Aeroelastic Wing project using flight test data is shown for several flight conditions (Mach numbers) by identifying a parsimonious system description with a high percent fit for cross-validated data.

  3. Aeroelastic Model Structure Computation for Envelope Expansion

    NASA Technical Reports Server (NTRS)

    Kukreja, Sunil L.

    2007-01-01

    Structure detection is a procedure for selecting a subset of candidate terms, from a full model description, that best describes the observed output. This is a necessary procedure to compute an efficient system description which may afford greater insight into the functionality of the system or a simpler controller design. Structure computation as a tool for black-box modelling may be of critical importance in the development of robust, parsimonious models for the flight-test community. Moreover, this approach may lead to efficient strategies for rapid envelope expansion which may save significant development time and costs. In this study, a least absolute shrinkage and selection operator (LASSO) technique is investigated for computing efficient model descriptions of nonlinear aeroelastic systems. The LASSO minimises the residual sum of squares by the addition of an l(sub 1) penalty term on the parameter vector of the traditional 2 minimisation problem. Its use for structure detection is a natural extension of this constrained minimisation approach to pseudolinear regression problems which produces some model parameters that are exactly zero and, therefore, yields a parsimonious system description. Applicability of this technique for model structure computation for the F/A-18 Active Aeroelastic Wing using flight test data is shown for several flight conditions (Mach numbers) by identifying a parsimonious system description with a high percent fit for cross-validated data.

  4. Real-Time Flight Envelope Monitoring System

    NASA Technical Reports Server (NTRS)

    Kerho, Michael; Bragg, Michael B.; Ansell, Phillip J.

    2012-01-01

    The objective of this effort was to show that real-time aircraft control-surface hinge-moment information could be used to provide a robust and reliable prediction of vehicle performance and control authority degradation. For a given airfoil section with a control surface -- be it a wing with an aileron, rudder, or elevator -- the control-surface hinge moment is sensitive to the aerodynamic characteristics of the section. As a result, changes in the aerodynamics of the section due to angle-of-attack or environmental effects such as icing, heavy rain, surface contaminants, bird strikes, or battle damage will affect the control surface hinge moment. These changes include both the magnitude of the hinge moment and its sign in a time-averaged sense, and the variation of the hinge moment with time. The current program attempts to take the real-time hinge moment information from the aircraft control surfaces and develop a system to predict aircraft envelope boundaries across a range of conditions, alerting the flight crew to reductions in aircraft controllability and flight boundaries.

  5. Critical point analysis of phase envelope diagram

    SciTech Connect

    Soetikno, Darmadi; Siagian, Ucok W. R.; Kusdiantara, Rudy Puspita, Dila Sidarto, Kuntjoro A. Soewono, Edy; Gunawan, Agus Y.

    2014-03-24

    Phase diagram or phase envelope is a relation between temperature and pressure that shows the condition of equilibria between the different phases of chemical compounds, mixture of compounds, and solutions. Phase diagram is an important issue in chemical thermodynamics and hydrocarbon reservoir. It is very useful for process simulation, hydrocarbon reactor design, and petroleum engineering studies. It is constructed from the bubble line, dew line, and critical point. Bubble line and dew line are composed of bubble points and dew points, respectively. Bubble point is the first point at which the gas is formed when a liquid is heated. Meanwhile, dew point is the first point where the liquid is formed when the gas is cooled. Critical point is the point where all of the properties of gases and liquids are equal, such as temperature, pressure, amount of substance, and others. Critical point is very useful in fuel processing and dissolution of certain chemicals. Here in this paper, we will show the critical point analytically. Then, it will be compared with numerical calculations of Peng-Robinson equation by using Newton-Raphson method. As case studies, several hydrocarbon mixtures are simulated using by Matlab.

  6. Critical point analysis of phase envelope diagram

    NASA Astrophysics Data System (ADS)

    Soetikno, Darmadi; Kusdiantara, Rudy; Puspita, Dila; Sidarto, Kuntjoro A.; Siagian, Ucok W. R.; Soewono, Edy; Gunawan, Agus Y.

    2014-03-01

    Phase diagram or phase envelope is a relation between temperature and pressure that shows the condition of equilibria between the different phases of chemical compounds, mixture of compounds, and solutions. Phase diagram is an important issue in chemical thermodynamics and hydrocarbon reservoir. It is very useful for process simulation, hydrocarbon reactor design, and petroleum engineering studies. It is constructed from the bubble line, dew line, and critical point. Bubble line and dew line are composed of bubble points and dew points, respectively. Bubble point is the first point at which the gas is formed when a liquid is heated. Meanwhile, dew point is the first point where the liquid is formed when the gas is cooled. Critical point is the point where all of the properties of gases and liquids are equal, such as temperature, pressure, amount of substance, and others. Critical point is very useful in fuel processing and dissolution of certain chemicals. Here in this paper, we will show the critical point analytically. Then, it will be compared with numerical calculations of Peng-Robinson equation by using Newton-Raphson method. As case studies, several hydrocarbon mixtures are simulated using by Matlab.

  7. Discriminating Dysarthria Type From Envelope Modulation Spectra

    PubMed Central

    Liss, Julie M.; LeGendre, Sue; Lotto, Andrew J.

    2013-01-01

    Purpose Previous research demonstrated the ability of temporally based rhythm metrics to distinguish among dysarthrias with different prosodic deficit profiles (J. M. Liss et al., 2009). The authors examined whether comparable results could be obtained by an automated analysis of speech envelope modulation spectra (EMS), which quantifies the rhythmicity of speech within specified frequency bands. Method EMS was conducted on sentences produced by 43 speakers with 1 of 4 types of dysarthria and healthy controls. The EMS consisted of the spectra of the slow-rate (up to 10 Hz) amplitude modulations of the full signal and 7 octave bands ranging in center frequency from 125 to 8000 Hz. Six variables were calculated for each band relating to peak frequency and amplitude and relative energy above, below, and in the region of 4 Hz. Discriminant function analyses (DFA) determined which sets of predictor variables best discriminated between and among groups. Results Each of 6 DFAs identified 2–6 of the 48 predictor variables. These variables achieved 84%–100% classification accuracy for group membership. Conclusions Dysarthrias can be characterized by quantifiable temporal patterns in acoustic output. Because EMS analysis is automated and requires no editing or linguistic assumptions, it shows promise as a clinical and research tool. PMID:20643800

  8. Revisiting the envelope approximation: Gravitational waves from bubble collisions

    NASA Astrophysics Data System (ADS)

    Weir, David J.

    2016-06-01

    We study the envelope approximation and its applicability to first-order phase transitions in the early Universe. We demonstrate that the power laws seen in previous studies exist independently of the nucleation rate. We also compare the envelope approximation prediction to results from large-scale phase transition simulations. For phase transitions where the contribution to gravitational waves from scalar fields dominates over that from the coupled plasma of light particles, the envelope approximation is in agreement, giving a power spectrum of the same form and order of magnitude. In all other cases the form and amplitude of the gravitational wave power spectrum is markedly different and new techniques are required.

  9. Measurement methods for evaluation of thermal integrity of building envelopes

    SciTech Connect

    Grot, R.A.; Burch, D.M.; Silberstein, S.; Galowin, L.S.

    1982-11-01

    This report presents reviews of various measurement and inspection techniques appropriate for the development of detailed diagnostic procedure for assessing the thermal performance of the exterior envelopes of federal buildings. The inspection techniques include the use of ground-based infrared thermographic surveys, aerial infrared surveys, tracer gas air infiltration measurement, pressurization tests for measuring the tightness of the building envelope, and spot radiometer surveys for detecting gross defects. Heat flow meters, a portable calorimeter, and a microprocessor-driven envelope testing unit are also considered.

  10. Carrier-envelope-phase stabilization via dual wavelength pumping.

    PubMed

    Seidel, Marcus; Brons, Jonathan; Lücking, Fabian; Pervak, Vladimir; Apolonski, Alexander; Udem, Thomas; Pronin, Oleg

    2016-04-15

    A power-scalable concept for carrier-envelope-phase stabilization is presented. It takes advantage of simultaneous pumping of the zero- and first-phonon absorption line of Yb:YAG at 969 and 940 nm. The concept was implemented to lock the carrier-envelope-offset frequency of a 45 W average power Kerr-lens mode-locked thin-disk oscillator. The lock performance is compared to previous experiments where carrier-envelope-stabilization was realized by means of cavity loss modulation.

  11. Survival of Enveloped and Non-Enveloped Viruses on Inanimate Surfaces

    PubMed Central

    Firquet, Swan; Beaujard, Sophie; Lobert, Pierre-Emmanuel; Sané, Famara; Caloone, Delphine; Izard, Daniel; Hober, Didier

    2015-01-01

    In the present study, we evaluated the viability of non-enveloped viruses, minute virus of mice (MVM) and coxsackievirus B4 (CVB4), and enveloped-viruses, influenza A virus (H1N1) and herpes simplex virus type 1 (HSV-1), on surfaces. We also investigated the impact of the initial concentration of proteins and sodium chloride on the persistence of infectious CVB4 on surfaces. Viral suspensions (>104.5 TCID50) were applied to petri dish lids and dried under the air flow of a biosafety cabinet. The recovered viral preparations were titered on appropriate cell lines. Enveloped viruses persisted for less than 5 days while CVB4 and MVM persisted for weeks. However, repetitive cycles of drying and resuspension had a stronger virucidal effect on CVB4 than on H1N1 and HSV-1. These repetitive cycles had no effect on the infectious titer of MVM. When exposed to drying, the initial concentrations of bovine serum albumin (from 0 to 90 mg mL−1), fetal calf serum (from 0 to 100%), and sodium chloride (from 0 to 300 mg mL−1) affected the viability of CVB4. CVB4 was more likely to be inactivated by drying in a protein-rich medium, whereas the impact of drying was reduced in the presence of sodium chloride. The results of the present study demonstrated that the resistance of viruses to drying, as suggested by iterative drying, was not due to the heterogeneity of viral subpopulations, but was influenced by media compositions and component concentrations, as illustrated in the model of CVB4. PMID:25843687

  12. ULTRASTRUCTURE AND PERMEABILITY OF NUCLEAR MEMBRANES

    PubMed Central

    Wiener, Joseph; Spiro, David; Loewenstein, Werner R.

    1965-01-01

    The fine structures of nuclear envelopes known to have different permeability properties were compared. Membranes of salivary gland cell nuclei of Drosophila (third instar) and Chironomus (prepupae), which are strong barriers to ion diffusion, and membranes of oocyte nuclei (germinal vesicle) of Xenopus and Triturus, which are much more ion-permeable, show no essential difference in size, frequency, and distribution of their membrane gaps ("pores") which could account for the marked disparities in membrane permeability. The gaps are occupied by diffuse electron-opaque material with occasional central regions of strong opacity. This material may possibly account for the high diffusion resistance of Drosophila and Chironomus nuclear envelopes, where the resistance is far too great to allow free diffusion through the gaps. But material of this kind is also present in the more permeable nuclear envelopes of Xenopus and Triturus oocytes, and there are no convincing structural differences discernible with the techniques employed. PMID:5892850

  13. EWS-FLI1 impairs aryl hydrocarbon receptor activation by blocking tryptophan breakdown via the kynurenine pathway.

    PubMed

    Mutz, Cornelia N; Schwentner, Raphaela; Kauer, Maximilian O; Katschnig, Anna M; Kromp, Florian; Aryee, Dave N T; Erhardt, Sophie; Goiny, Michel; Alonso, Javier; Fuchs, Dietmar; Kovar, Heinrich

    2016-07-01

    Ewing sarcoma (ES) is an aggressive pediatric tumor driven by the fusion protein EWS-FLI1. We report that EWS-FLI1 suppresses TDO2-mediated tryptophan (TRP) breakdown in ES cells. Gene expression and metabolite analyses reveal an EWS-FLI1-dependent regulation of TRP metabolism. TRP consumption increased in the absence of EWS-FLI1, resulting in kynurenine and kynurenic acid accumulation, both aryl hydrocarbon receptor (AHR) ligands. Activated AHR binds to the promoter region of target genes. We demonstrate that EWS-FLI1 knockdown results in AHR nuclear translocation and activation. Our data suggest that EWS-FLI1 suppresses autocrine AHR signaling by inhibiting TDO2-catalyzed TRP breakdown.

  14. Experimental study of vortex breakdown in swirling jets

    NASA Astrophysics Data System (ADS)

    Billant, Paul; Chomaz, Jean-Marc; Huerre, Patrick

    1998-12-01

    The goal of this study is to characterize the various breakdown states taking place in a swirling water jet as the swirl ratio S and Reynolds number Re are varied. A pressure-driven water jet discharges into a large tank, swirl being imparted by means of a motor which sets into rotation a honeycomb within a settling chamber. The experiments are conducted for two distinct jet diameters by varying the swirl ratio S while maintaining the Reynolds number Re fixed in the range 300Breakdown is observed to occur when S reaches a well defined threshold Sc[approximate]1.3 1.4 which is independent of Re and nozzle diameter used. This critical value is found to be in good agreement with a simple criterion derived in the same spirit as the first stage of Escudier & Keller's (1983) theory. Four distinct forms of vortex breakdown are identified: the well documented bubble state, a new cone configuration in which the vortex takes the form of an open conical sheet, and two associated asymmetric bubble and asymmetric cone states, which are only observed at large Reynolds numbers. The two latter configurations differ from the former by the precession of the stagnation point around the jet axis in a co-rotating direction with respect to the upstream vortex flow. The two flow configurations, bubble or cone, are observed to coexist above the threshold Sc at the same values of the Reynolds number Re and swirl parameter S. The selection of breakdown state is extremely sensitive to small temperature inhomogeneities present in the apparatus. When S reaches Sc, breakdown gradually sets in, a stagnation point appearing in the downstream turbulent region of the flow and slowly moving upstream until it reaches an equilibrium location. In an intermediate range of Reynolds numbers, the breakdown threshold displays hysteresis lying in the ability of the breakdown state to remain stable for Sbreakdown, i.e. when 0

  15. Quenching of vortex breakdown oscillations via harmonic modulation

    NASA Astrophysics Data System (ADS)

    Lopez, J. M.; Cui, Y. D.; Marques, F.; Lim, T. T.

    Vortex breakdown is a phenomenon inherent to many practical problems, such as leading-edge vortices on aircraft, atmospheric tornadoes, and flame-holders in combustion devices. The breakdown of these vortices is associated with the stagnation of the axial velocity on the vortex axis and the development of a near-axis recirculation zone. For large enough Reynolds number, the breakdown can be time-dependent. The unsteadiness can have serious consequences in some applications, such as tail-buffeting in aircraft flying at high angles of attack. There has been much interest in controlling the vortex breakdown phenomenon, but most efforts have focused on either shifting the threshold for the onset of steady breakdown or altering the spatial location of the recirculation zone. There has been much less attention paid to the problem of controlling unsteady vortex breakdown. Here we present results from a combined experimental and numerical investigation of vortex breakdown in an enclosed cylinder in which low-amplitude modulations of the rotating endwall that sets up the vortex are used as an open-loop control. As expected, for very low amplitudes of the modulation, variation of the modulation frequency reveals typical resonance tongues and frequency locking, so that the open-loop control allows us to drive the unsteady vortex breakdown to a prescribed periodicity within the resonance regions. For modulation amplitudes above a critical level that depends on the modulation frequency (but still very low), the result is a periodic state synchronous with the forcing frequency over an extensive range of forcing frequencies. Of particular interest is the spatial form of this forced periodic state: for modulation frequencies less than about twice the natural frequency of the unsteady breakdown, the oscillations of the near-axis recirculation zone are amplified, whereas for modulation frequencies larger than about twice the natural frequency the oscillations of the recirculation

  16. 42GHz ECRH assisted Plasma Breakdown in tokamak SST-1

    NASA Astrophysics Data System (ADS)

    Shukla, B. K.; Pradhan, S.; Patel, Paresh; Babu, Rajan; Patel, Jatin; Patel, Harshida; Dhorajia, Pragnesh; Tanna, V.; Atrey, P. K.; Manchanda, R.; Gupta, Manoj; Joisa, Shankar; Gupta, C. N.; Danial, Raju; Singh, Prashant; Jha, R.; Bora, D.

    2015-03-01

    In SST-1, 42GHz ECRH system has been commissioned to carry out breakdown and heating experiments at 0.75T and 1.5T operating toroidal magnetic fields. The 42GHz ECRH system consists of high power microwave source Gyrotron capable to deliver 500kW microwave power for 500ms duration, approximately 20 meter long transmission line and a mirror based launcher. The ECRH power in fundamental O-mode & second harmonic X-mode is launched from low field side (radial port) of the tokamak. At 0.75T operation, approximately 300 kW ECH power is launched in second harmonic X-mode and successful ECRH assisted breakdown is achieved at low loop_voltage ~ 3V. The ECRH power is launched around 45ms prior to loop voltage. The hydrogen pressure in tokamak is maintained ~ 1×10-5mbar and the pre-ionized density is ~ 4×1012/cc. At 1.5T operating toroidal magnetic field, the ECH power is launched in fundamental O-mode. The ECH power at fundamental harmonic is varied from 100 kW to 250 kW and successful breakdown is achieved in all ECRH shots. In fundamental harmonic there is no delay in breakdown while at second harmonic ~ 40ms delay is observed, which is normal in case of second harmonic ECRH assisted breakdown.

  17. Aerosol-induced laser breakdown thresholds: wavelength dependence.

    PubMed

    Pinnick, R G; Chylek, P; Jarzembski, M; Creegan, E; Srivastava, V; Fernandez, G; Pendleton, J D; Biswas, A

    1988-03-01

    Aerosol-induced loser breakdown thresholds have been measured for liquid droplets at wavelengths lambda= 1.064, 0.532, 0.355, 0.266 microm using a Nd:YAG laser with 5-10-ns pulse duration. Breakdown thresholds are 2-3 orders of magnitude below those for clean air and range from 4 x 10(7) to 3 x 10(9) W cm(-2) for nominal 50-microm diam droplets, depending on laser wavelength and droplet composition. Thresholds decrease with decreasing wavelength; they also decrease for droplets having a higher real refractive index. For water droplets the breakdown threshold intensity varies approximately as lambda(0.5). The wavelength dependence of breakdown thresholds can be qualitatively explained by considering (1) the effect of enhancement of internal fields and energy density within and near droplets and (2) the increasing importance of multiphoton absorption processes at shorter wavelengths. Laser transmission losses through the breakdown plasma and observations of the suppression of stimulated Raman scattering by the addition of small quantitites of absorbing material to water and carbon tetrachloride droplets are also reported.

  18. Detectable initial breakdown pulses and the following discharges

    NASA Astrophysics Data System (ADS)

    Zhang, Y.; Lu, W.; Zheng, D.

    2014-12-01

    The initial breakdown which was descripted by so-called BIL model is an important discharge process. Bipolar pulses as important signals of initial breakdown process, were usually used in the research. However, the detectability of the pulses is different in different researches. At the same time, the discharge process from initial breakdown to return stroke is not always the simple BIL model. In the research, using electric field waveforms of lightning discharges observed in Beijing and Guangzhou, the detectable initial breakdown (preliminary breakdown, PB) pulses and the following discharges are analyzed. The results show that, (1) Although, the percentage of detectable PB pulse trains is just as a function of latitude in large range of latitude, the values are similar in Beijing and Guangzhou, which indicate that the characteristic of storm also play a key role in detectable PB besides the latitude. (2)According to the difference in discharge process of following PB pulse, three discharge types, which exhibits PB pulses followed by inverted IC, hybrid flash, leader and RS, can be identified. And the fourth type only exhibits a RS waveform without detectable PB pulses. (3)The discharge with undetectable PB pulse is dominant in Beijing and Guangzhou. But the percentage of each discharge type is obvious different in the two regions. The occurrence of different discharge types and different percentage in Beijing and Guangzhou can be contributed to the difference in low positive charge region.

  19. Delta Wing Vortex Breakdown Suppression by Vortex Core Oscillation

    NASA Astrophysics Data System (ADS)

    Cain, Charles

    2000-11-01

    The flow over a delta wing is characterized by two counter-rotating vortices that can undergo a sudden radial expansion at high angles of attack known as vortex breakdown. Downstream of this breakdown is a region of organized unsteady flow that can cause tail buffeting and structural fatigue, especially on twin-tailed aircraft. The recent self-induction theory of vortex breakdown points to the "pile-up" of vorticity due to the linear addition of vorticity in the spiraling shear layer that surrounds the vortex core as a principal cause of vortex breakdown (Kurosaka 1998). Based on that theory, this research attempts to relieve vorticity pile-up by altering the straight-line path of the vortex core and preventing the linear addition of vorticity. This is accomplished by applying a combination of periodic blowing and suction with low mass and momentum flux. The blowing and suction are directed normal to the low-pressure surface and supplied from ports under the vortex core which are near the forward tip of the delta wing. This oscillating input causes the vortex core to transition into a spiral formation downstream of the input ports. Initial results indicate that this change in the vortex core path may prevent vortex breakdown over the surface of the delta wing.

  20. Chlorophyll breakdown and chlorophyll catabolites in leaves and fruit†

    PubMed Central

    Kräutler, Bernhard

    2010-01-01

    Chlorophyll metabolism probably is the most visible manifestation of life. Total annual turnover of chlorophyll has been estimated to involve more than 1000 million tons. Surprisingly, chlorophyll catabolism has remained an enigma until less than twenty years ago, when a colorless chlorophyll catabolite from senescent plant leaves was identified and its structure was elucidated. In the meantime, chlorophyll breakdown products have been identified in a variety of plant leaves and their structural features have been elucidated. Most recently, chlorophyll breakdown products have also been identified in some ripening fruit. Chlorophyll breakdown in vascular plants only fleetingly involves enzyme-bound colored intermediates. The stage of fluorescent catabolites is also passed rapidly, as these isomerize further to colorless nonfluorescent tetrapyrrolic catabolites. The latter accumulate in the vacuoles of de-greened leaves and are considered the final products of controlled chlorophyll breakdown. The same tetrapyrroles are also found in ripening fruit and are effective antioxidants. Chlorophyll breakdown leads to tetrapyrroles that appear to have physiologically beneficial chemical properties, and it may thus not merely be a detoxification process. PMID:18846275